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PP3 - Displayed on Tuesday, September 11 Gynecopathology PP3-1 BILATERAL SERTOLI -LEYDIG TUMOR OF THE OVARY WITH RETIFORM PATTERN Amel Trabelsi, Sarra Mestiri, Atef Ben Abdelkader, Med. Tahar Yacoubi, Badreddine Sriha, Sassi Bouguizéne, Sadik Korbi Pathology Department, F. Hached Hospital, 4000 Sousse, Tunisia Background: Sertoli-Leydig tumor of the ovary is rare, representing less then 1 % of all ovarian tumors. The mean age of patients is usually 25 years, and is usually unilateral. The retiform pattern is seen in young patients and malignancy depend on differentiation at histopathological examination. Method: We report a case of 35 year-old woman that consulted in emergency for an acute abdominal pain. Results: Radiological investigations showed a right ovarian tumor. The patient underwent a right ovariectomy with biopsy of the peritoneum. Histological examination concluded to a Sertoli-Leydig tumor with a retiform pattern; One year later, the patient presented with a left ovarian mass. Microscopically, the mass was also a Sertoli-Leydig tumor with a retiform pattern. Clinical outcome was favourable with no local recurrence two years after. Conclusion: Sertoli-Leydig tumor of the ovary is rare and is usually unilateral; malignancy is based on the degree of differentiation at histopathological examination. PP3-2 PRIMITIVE NEUROECTODERMAL TUMOR OF THE CERVIX: A CASE REPORT Amel Trabelsi, Sarra Mestiri, Lilia Ben Yacoub-Abid, Fatma Kebir, Soumaya Rammeh, Moncef Mokni, Atef Ben Abdelkader, Sadok Korbi Service de Pathologie, Hopital F. Hached, 4000 Sousse, Tunisia Background: Primitive neuroectodermal tumor of the cervix is extremely rare in this location, raising for the pathologist differential diagnosis difficulties. Immunohistochemistry is of great value to lead to the correct diagnosis. Method: We report a case of 50 year-old menopausal Tunisian woman who presented with vaginal bleeding. Physical examination revealed a 2,5 cm cervical exophytic mass. Results: The specimen biopsy showed a round small cell malignant proliferation. Diagnosis of primitive neuroectodermal tumour of the cervix was retained after immunohistochemical results (positivity of mic-2 : CD 99). The patient underwent a radical surgical treatment followed by adjuvant chemotherapy. Clinical outcome was marked by local recurrence 18 months later and the patient died 6 months later. Conclusion: Primitive neuroectodermal tumor of the cervix is uncommon, raising diagnostic difficulties. Its prognosis is usually unfavourable. PP3-3 OVEREXPRESSION OF LAMININ-5 GAMMA 2 CHAIN IN CLEAR CELL CARCINOMA OF THE OVARY Noriko Kato, Teiichi Motoyama Department of Pathology, Yamagata University School of Medicine, Japan Background: One of the characteristic microscopic features of ovarian clear cell carcinoma (CCC) is the densely hyaline basement membrane material expanding the stroma. The biological significance of this material, however, has remained unclear. Recent studies have shown that laminin-5 (LN-5), a major component of the epithelial basement membrane, plays a more active role in cell migration or tumor invasion. Method: Twenty-five surgically resected CCCs were examined for LN-5 expression immunohistochemically, using an antibody against

LN-5 gamma 2 chain. For comparison, 5 borderline clear cell tumors, 10 serous adenocarcinomas, and 10 endometrioid adenocarcinomas were examined. Three CCC cell lines were analyzed in vitro for migration over excessive recombinant LN-5, with or without function-blocking antibody against integrin Į3. Results: All of the 25 CCCs showed a focal or diffuse immunoreactivity with the LN-5 gamma 2 chain in the tumor stroma; whereas, borderline clear cell tumors, serous or endometrioid adenocarcinomas rarely showed a stromal immunoreactivity. Cytoplasmic accumulation of the LN-5 gamma 2 chain was far less common than stromal accumulation, suggesting an accelerated secretion in CCC. In vitro, CCC cell lines showed a significant increase of cell migration over excessive LN-5, and the migration was blocked by an antibody against integrin Į3. Conclusion: The stromal accumulation of the LN-5 gamma 2 chain is common in ovarian CCC but rare in borderline clear cell tumors or other ovarian carcinomas. It is indicated that an interaction between CCC cells and extracellularly accumulated LN-5 is responsible for cell migration and the subsequent stromal invasion of CCC. PP3-4 REDUCED EXPRESSION OF THE METASTASIS SUPPRESSOR KAI-1 IN UTERINE TUMORS OF EPITHELIAL AND STROMAL ORIGIN: CORRELATION WITH P53 STATUS Juliane Nowak1, Christoph M. Bamberger2, Thomas Loning3, Maria Sajin4, Karin Milde-Langosch5, Sylvia L Asa6, Ana-Maria Bamberger2 1 Department of Pathology, University of Toronto, Canada 2 Section on Endocrinology and Metabolism of Ageing, University Clinic Hamburg - Eppendorf, Hamburg, Germany 3 Department of Pathology, Centre for Cytology and Gynecopathology, Germany 4 Department of Morphopathology, University of Bucharest, Bucharest, Romania 5 Department of Gynecology, University Clinic HamburgEppendorf, Hamburg, Germany 6 Department of Pathology, University of Toronto, Canada Background: The ability of tumor cells to metastasize often requires additional genetic changes, which result in activation and inactivation of metastasis stimulating and metastasis suppressor genes. Recently KAI1 has emerged as a tumor-suppressor and more specifically as a metastasis suppressor. A loss of KAI-1 expression is associated with the advanced stages of many human malignancies and results in the acquisition of invasive and metastatic capabilities by tumor cells. The underlying mechanisms responsible for this down-regulation of KAI-1 expression remain to be resolved. Method: In the present study, immunohistochemistry and Western blot analysis were performed on uterine tumors, hyperplasias and normal endometrium to investigate the expression pattern and cell-type specific localization of KAI-1 and to correlate it with the expression of p53 and histological and clinical data. Result: 29 (69.04%) of 42 endometrial carcinomas showed reduced or absent KAI-1 expression, which correlated with strong expression of p53 (P < 0.001). 13 tumors (30.95%) of 42 demonstrated a moderate score (4-7) of Kai-1 expression. It is interesting that these carcinomas were all well-differentiated tumors of malignancy grade G1/G2, which showed low expression of p53. There were significant correlations between KAI-1 expression and histological type, tumor grading and clinical stage (P < 0.001). Most of the investigated sarcomas of the uterus (including 5 carcinosarcomas, 8 endometrial stromal sarcomas and 2 leiomyosarcomas) were clearly negative for KAI-1, whereas these tumors displayed a strong immunostaining for p53. Conclusion: In conclusion, our results demonstrate a strong inverse correlation between KAI-1 and p53 expression and it thus seems, that reduced KAI-1 expression possibly as a result of disregulated/mutated p53 gene could be an important step in uterine carcinogenesis.

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PP3-5 COMPARION OF MICROIMMUNOFLUORECENCE, ELISA, RAPID DETECTION KIT (DIMA) AND GIMENEZ STAINING FOR DETECTION OF CHLAMYDIA INDUCED CERVICITIS Nour Amirmozafari, Homa Forohesh, Liella Gang Iran University of Medical Sciences, School of Medicine, Tehran, Iran

positive predictive value was 96,8 % for benign tumors, 85,7 % for borderline tumors and 100 % for malignant tumors. Conclusion: Frozen section diagnosis seems to be a reliable method for the surgical management of patients with ovarian masses with a high sensitivity for malignant tumors and a low false-positive rate. However, diagnostic problems can occur in mucinous and borderline tumors wich require extensive sampling.

Backgroud: Chlamydia trachomatis is one of the most prevalent causative agent of STD. It causes a variety of genital tract complications such as urethritis, cervicitis, endometritis, epididymitis, and lymphogranuloma venereum. The prevalence rate of Chlamydia cervicitis is varied in different societies. In a recent study, the rate in sexually active American women was in the range of 5-15%. The purpose of this study was to evaluate the prevalence rate of Chlamydia induced cervicitis in Iranian women, and to compare the efficacy of different detection techniques. Method: A total of 137 women with cervicitis were admitted for this study. Two endocervical swabs were obtained. One was used for the rapid Dima test, and the other swab was subjected to Gimenez staining. Blood samples were also obtained for serological tests. Results: ELISA indicated that 18 patients had positive IgG antibody levels in their blood, and 4 of them had IgM antibodies against Chlamydia trachomatis. Ten patients had significant IgG levels and 3 of them had anti-Chlamydia IgM according to the MIF test results. Dima rapid detection test was able to show positive results for only 5 patients. We were not able to detect any Chlamydia inclusion bodies with direct microscopy after Gimenez staining. Conclusion: The MIF technique which is generally considered to be the Gold Standard serological detection method, showed the prevalence rate of Chlamydia cervicitis to be 7.2%. There was no statistically significant difference in IgM titers detected by ELISA and MIF methods. But there was statistically significant different IgG titer rates between ELISA and MIF. Therefore, it is suggested that any ELISA positive IgG titer samples be rechecked and reconfirmed by MIF. Due to the lack of any Chlamydia inclusion body detection by direct microscopy, Gimenez staining is not recommended as a diagnostic tool.

PP3-7 OVEREXPRESSION OF COX-2 AND HER-2/NEU IN PAGETS´S DISEAE OF THE VULVA AND THE BREAST Lars-Christian Horn1, Sandra Purz1, Christine Krumpe1, Karl Bilek2 1 University of Leipzig, Department of Gynecologic & Perinatal Pathology, Germany 2 University of Leipzig, Department of Obstetrics & Gynecology, Germany

PP3-6 ACCURACY AND RELIABILITY OF FROZEN SECTION DIAGNOSIS IN A SERIES OF 183 OVARIAN TUMORS Farah-Klibi Faten Charles Nicolles Hospital Tunisia

PP3-8 HIGH FREQUENCY OF LYMPHOVASCULAR INVASION IN EARLY ADENOCARCINOMA OF THE UTERINE CERVIX Keiko Abe1, Chikako Suzuki2, Atsushi Arakawa2, Masaaki Suzuki3, Toshiharu Matsumoto2 1 Department of Humanpathology, Juntendo University School of Medicine, Sanikukai Hospital, Tokyo, Japan 2 Department of Obstetrics and Gynecology, Juntendo University School of Medicine, Japan 3 Sanikukai Hospital, Tokyo, Japan

Objective: Frozen section is an important diagnostic tool in determining the nature of ovarian masses. However, it still has some pitfalls. This retrospective study was conducted to determine the accuracy of frozen section diagnosis of ovarian masses and to discuss the type and the reasons of inaccuracies associated with this procedure. Methods: From January 2002 to April 2005, 183 ovarian specimens were examined by frozen section. The frozen section results were compared with the final diagnosis in paraffin sections. The sensitivity (Se), specificity (Sp) and predictive values of frozen section diagnosis were calculated. The 95 % confidence interval of each parameter was determined. Results: Ovarian masses accounted for 11,1 % of all specimens examined extemporaneously. The frozen section diagnosis was benign in 85,8 % of the cases, borderline in 3,8 % and malignant in 7,7 %. The frozen section diagnosis was deffered to permanent sections in 2,7 % of all cases. Overall diagnostic agreement was 96,6 % (Kappa=0,85). Six cases (3,4 %) were incorrectly diagnosed by frozen section. There was one false-positive and five false-negative cases. All of them were due to erroneous interpretation. The majority of the cases of disagreement were mucinous and borderline tumors. The Se for benign, borderline and malignant tumors were 99,3 %, 66,7 % and 87,5 % respectively. The Sp for benign tumors was 80 %, for borderline tumors 99,4 % and for malignant tumors 100 %. The

Background: Paget´s disease (PD) of the breast as well as the vulva is a rare condition that accounts for about 4% of breast neoplasms and 1% of vulvar malignancies. Recurrent disease after breast and vulvar surgery might be a challenge. To evaluate therapeutically relevant molecules, tissue from mammary and vulvar PD lesions was investigated immunohistochemically. Methods: Histopathologic samples from eleven patients with mammary PD and eight patients with vulvar PD were stained with antibodies against estrogen and progesterone receptors, HER-2/neu and COX-2 followed by semiquantitative evaluation of the staining results. Results: All tested mammary lesions as well as seven out of eight vulvar PD were negative for estrogen and progesterone receptor. Strong membranous staining for HER2/neu (Score 3) was seen in all cases. Six out of eight vulvar and ten out of eleven mammary PD showed COX-2 overexpression. Conclusions: PD of the breast and the vulva is not under hormonal control of estrogens and progesterons, therefore, antihormonal therapy is not indicated. The high frequency of Her2/neu and COX-2 overexpression, however, suggests that these molecules could be therapeutically relevant in patients with PD. These results require further investigation.

Abstract Backgrounds: The prognostic factors of adenocarcinoma of the uterine cervix were said to be the stage of disease and lymph node metastasis. The incidence of lymph node metastasis was reported to be significantly higher in the patients with adenocarcinoma than in those with squamous cell carcinoma of the uterine cervix even in stage Ib. The other report said that the risk of nodal metastases is higher in the patients with cervical adenocarcinoma showing stromal invasion over 5mm in depth. These studies suggested that lymphovascular involvement increased in the early stage of adenocarcinoma of the uterus. Methods: There were eight patients who underwent radical hysterectomy and bilateral salpingo-oophorectomy in 2000-2006 at Sanikukai Hospital in Tokyo. The average age of the patients was 47.1 years (36-75 in range). One or two specimen were selected for the following examinations. Each specimen was stained with Hematoxyline-Eosine and Elastica-van-Gieson. Immunohistochemical staining was also performed for anti-D240. Results: Histological type of the eight patients was four

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endometrioid type, two endocervical type and two adenocarcinoma in situ. Stages of eight patients with cervical adenocarcinoma showed that 4 cases of stage Ib2, two cases of stage Ib1 and two cases of adenocarcinoma in situ. Three cases of four Ib2 cases showed prominent lymphatic invasion. Two of the four Ib2 cases revealed vascular invasion. Lymph vessel invasion was found on the peripheral area of the tumor, with the background of markedly lymph vessel proliferation. On the other hand, both stages of Ib1 and adenocarcinoma in situ did not show any lymphovascular involvement. No lymph nodes metastasis was found in all of the eight cases. Conclusion: Lymphovascular invasion was frequently observed in adenocarcinoma of the uterine cervix stage Ib2 even in the condition of no lymph nodes metastasis. PP3-9 LYMPHANGIOLEIOMYOMATOSIS IN THE FEMALE GENITAL TRACT Takuo Hayashi1, Toshio Kumasaka1, Keiko Mitani1, Yasuka Miyakuni1, Yoko Gunji2, Koichi Suda1, Kuniaki Seyama2 1 Department of Humanpathology of Juntendo University School of Medicine, Japan 2 Department of Respiratory Medicine of Juntendo University School of Medicine, Japan Background: Lymphangioleiomyomatosis (LAM) is a rare, multisystem disorder of unknown etiology that affects exclusively young women and characterized by proliferation of abnormal smooth muscle-like cells (LAM cells). LAM is usually recognized of its pulmonary manifestations characterized by cystic destruction of the lung parenchyma due to proliferation of LAM cells. LAM cells can also involve the extrapulmonary organs including the lymphatic system. However, through analysis of female genital tract in patients with LAM has not been reported. Method: Six cases with LAM involving the female genital tract (3 surgical specimens and 3 autopsy cases) were collected from archives of the Department of Humanpathology at Juntendo University. A uterus was cut from cervix to fundus to identify the LAM lesions (average, 26.4 specimens per case; range, 2-78). Adnexa were serially cut from isthmus to fimbria at 5-mm thickness, including ovary and mesovarium (average, 16.4 specimens per case; range, 2-30). Retroperitoneal lymph nodes were obtained from 5 cases. Immunohistochemical staining was carried out using monoclonal antibodies against Į-smooth muscle actin (Į-SMA, dilution 1:200, Dako Cytomation, Carpinteria, CA), HMB45 (dilution 1:50, Dako Cytomation), Flt-4 (VEGFR3,dilution 1:50,R&D Co Ltd., Minneapolis, MN), and CD10 (dilution 1㧦200, Novocastra Laboratories Ltd., New Castle, UK ). Results: 1) Patients’ age ranged from 25 to 83 years (mean 41.3 years) when the specimens were obtained. Two patients had tuberous sclerosis complex (TSC). 2) All cases showed LAM lesions in the adnexa and the retroperitoneal lymph nodes, while four of 5 cases did them in the uterus. 3) LAM lesions were detected in the serosa and/or subserosa of the uterus in all cases. However LAM lesions were not identified in the endometrium. 4) Proliferating patterns of LAM cells in the uterus were categorized into two types: nodular and diffuse type. All of sporadic LAM patients revealed nodular type and two TSC-LAM patients did diffuse type of uterine LAM lesions. 5) LAM cell clusters (LCCs) in the lymphatic vessels were detected in all cases. Conclusion: The anatomic distribution of LAM in the female genital tract is similar to that of metastatic cancers to the uterus, and LAM lesions are accompanied by LCCs in the lymphatic vessels in all cases. These results suggest that LAM lesions in the female genital tract may be disseminated from the extragenital tract through lymphatic vessels.

PP3-10 COX- 2, HER- 2/NEU AND HORMONE RECEPTOR ANALYSIS IN PRIMARY PAPILLARY CARCINOMA OF THE PERITONEUM Lars-Christian Horn1, Sandra Purz1, Michael Hockel2, Jens Einenkel2 1 Institute of Pathology, Division of Gynecologic and Perinatal Pathology, University of Leipzig, Germany 2 Department of Obstetrics and Gynecology, University of Leipzig, Germany Background: Primary peritoneal carcinomas (PPC) are rare but aggressive malignancies. Evaluating therapeutic relevant molecules PPC were investigated immunohistochemically. Methods: 26 PPC were stained with antibodies against estrogen and progesterone receptors, HER-2/neu and COX-2 with semiquantitative evaluation of the staining results. Results: Twenty one out of the 26 PPC (80.8%) were negative or showed only weak expression for estrogen receptor. One half of the tumors showed moderate or strong positivity for progesterone receptor. Only three PPC (11.5%) showed strong immunoreaction for HER-2/neu (Score 3). Nearly two thirds of the PPC represented COX-2 overexpression. Conclusions: PPCs are probably not under hormonal control of estrogens and lack HER2/neu overexpression in the majority of cases. So, anti-hormonal or antibody therapy with herceptine is not indicated. The use of COX-2 inhibitores might be helpful in PPC. PP3-11 ALTERATION OF CELL-CYCLE REGULATORY PROTEINS AND TUMOR BIOLOGY OF SMALL CELL NEUROENDOCRINE CARCINOMAS OF THE UTERINE CERVIX Lars-Christian Horn1, Jeanett Edelmann2, Bettina Hentschel3, Karl Bilek4, Uwe-Gerd Liebert5, Jens Einenkel4, Steffen Hauptmann6, Cornelia Leo4 1 Institute of Pathology, Division of Gynecopathology, University of Leipzig, Germany 2 Institute of Legal Medicine, University of Leipzig, Germany 3 Institute of Medical Informatics, Statistics and Epidemiology, Germany 4 Department of Obstetrics and Gynecology, University of Leipzig, Germany 5 Institute of Virology, University of Leipzig, Germany 6 Department of Pathology, University of Halle-Wittenberg, Germany Background: Small cell neuroendocrine carcinomas (SmCC) of the uterine cervix are rare tumors. The knowledge regarding protein-expression of several checkpoint candidates of cell-cycleregulation, their proliferative activity and the prognostic impact of only focal neuroendocrine differentiation is limited. So, SmCC were identified from our files and studied immunohistochemically with HPV-analysis. Methods: Immunohistochemical anlysis consisted neuroendocrine markers, p53, p16, p14. pRb-1, cyclin D1 and Ki-67. PCR-analysis, using general primers was performed for HPV-analysis. In cases with mixed tumors, the percentage of SmCC-component was calculated and correlated with survival. Results: Nine out of 677 tumors (1.3%) were classified as SmCC after Grimelius-staining and immunohistochemistry for NSE, chromogranin A, synaptophysin and CD 56. Two SmCC`s represented p53- and one case p14-positivity. Cyclin D1-staining was completely negative. All buat one SmCC were negative for pRb-1. All cases showed strong nuclear and/or cytoplasmic p16-immunostaining. Seven tumors represented HPV-positivity for high risk types. Four patients died of the tumor after a median time of 36.7 months (range 15 to 56 months), representing a 5-year-survivalrate of 56%. Even a SmCC-component of 17% was associated with fatal course in mixed cases. SmCC represented significant lower proliferation (Ki-67 labeling-index) than the non-small-cell

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component in the same tumor (12.8 vs. 70.8%; p<0.001). Conclusions: The overexpression of p16 in all cases but a loss of cyclin D1 and pRb1 in the majority of cases, might indicate a widespread disruption of the Rb-pathway which could be responsible for the aggressive course of SmCC. Even a small SmCC-component in mixed carcinoma of the uterine cervix was associated with adverse outcome. Proliferative activity, determined by Ki-67 labeling index, is of no prognostic value. PP3-12 ASSOCIATION OF P63 WITH TUMOR CELL DISSOCIATION AND JUXTATUMORAL STROMAL REMODELLING IN CERVICAL CARCINOMA Lars-Christian Horn1, Nicole Hommel1, Bettina Hentschel2, Karl Bilek3, Ulrike Roschlau1, Ulf-Dietrich Braumann4, Jens Einenkel3 1 Institute of Pathology, Division of Gynecologic Pathology University of Leipzig, Germany 2 Institute for Medical Informatics, Statistics and Epidemiology, University of Leipzig, Germany 3 Department of Obstetrics and Gynecology, University of Leipzig, Germany 4 Interdsiziplinary Centre of Bioinformatics, University of Leipzig, Germany Background: p63 (syn. KET and p51) is located at the short arm at chromosome 3 (3q27-29) with tumor suppressive and oncogenic properties. Its overexpression has been reported in carcinomas of the head and neck and the lungs. The present study evaluates p63-expression on carcinoma of the cervix uteri in correlation to tumor-stromal interaction. Methods: Paraffine embedded tumoral tissue from 140 patients with cervical carcinoma FIGO stage III and IV, treated with radiation therapy, were examined immunohistochemically. Staining results were evaluated using an immunoreactive score (staining intensity (1-3) x percentage of positive stained nuclei). The score values were compared to histologic tumor type, tumor grade, pattern of invasion and grade of juxtatumoral stromal remodelling (i.e. desmoplastic change). Results: Squamous cell carcinomas showed more often p63-expression than adenocarcinomas (p=0.0001). Poorly differentiated tumors (grade 3) represented a reduced p63 expression (p=0.001). Carcinomas with high tumor cell dissociation (characterised by spray-like pattern of invasion) and those with strong peritumoral stromal reaction were also associated with a loss of p63-expression (p<0.02 and p=0.074). There was no correlation between p63-expression and response to radiation therapy nor to overall survival. Conclusions: p63 is associated with high tumor cell dissociation and strong remodelling of juxtatumoral stroma in carcinoma of the cervix uteri. But, the mechanism how acts p63 in the context of the alteration of tumor cell adhesion and stromal remodelling is not well understood at time. PP3-13 PATTERN OF INVASION (TUMOR CELL DISSOCIATION) AS PROGNOSTIC FACTOR IN CARCINOMA OF THE UTERINE CERVIX Lars-Christian Horn1, Uta Fischer2, Bettina Hentschel3, Georgios Raptis1, Karl Bilek2, Christine E Richter1, Ulf-Dietrich Braumann4, Jens Einenkel2 1 Institute of Pathology, Division of Gynecologic Pathology, University of Leipzig, Germany 2 Department of Obstetrics and Gynecology, University of Leipzig, Germany 3 Institute for Medical Informatics, Statistics and Epidemiology, University of Leipzig, Germany 4 Interdisciplinary Centre of Bioinformatics, University of Leipzig, Germany Background: Different patterns of invasion (representing different grades of tumor cell dissociation) are associated with

prognostic outcome in cancer. We evaluated the prognostic value of different patterns of invasion (PI) in cervical carcinomas (CX). Methods: 611 surgically treated CX (FIGO IB to IIB) were reevaluated histologically regarding the PI, using a three-level scoring system. Closed PI was defined as cohesive growth with well-delineated (pushing) borders. In finger-like PI the tumor grows in solid cords/trabecles. Highly dissociative growth in small groups or single cells was defined as spray-like PI. Types of PI were correlated to tumor stage, histo-morphologic factors and prognostic outcome. Results: 60% of the tumors showed a spray-like PI, 30% a finger-like PI, and only 7.4% were of the closed type. Spray-like PI showed a significant correlation with advanced stage disease, lymphovascular space involvement, poorly differentiated tumors and pelvic lymph node metastases. Spray-like PI was accompanied by a reduced 5-year overall survival when compared to the finger-like and closed PI (68.7% vs. 80.9% vs. 88.5%; p=0.0004). The prognostic impact of the PI disappeared in node-positive patients (p=0.06), but persisted in patients without pelvic lymph node disease (p=0.03). In multivariate analysis, using COX-regression model, the PI represented as independent prognostic factor. Conclusions: Spray-like PI, (i.e. highest degree of tumor cell dissociation) is associated with advanced tumor stages, increased rate of recurrency and a reduced overall survival. In separate analysis of patients with and without lymph node metastases, the impact of PI persisted only in node-negative cases as an prognostic factor. PP3-14 SURVIVIN EXPRESSION IN THE MANAGEMENT OF ENDOMETRIAL CANCER PATIENTS Maria Lambropoulou1, Dimitrios Stefanou2, George Alexiadis1, Grigorios Tripsianis3, Demetrio Tamiolakis1, Ekaterini Chatzaki4, Vasilis Liberis5, Nikolas Papadopoulos1 1 Department of Histology-Embryology, Democritus University of Thrace, Greece 2 Department of Pathology, University of Ioannina, Greece 3 Department of Medical Statistics, Democritus University of Thrace, Greece 4 Department of Pharmacology, Democritus University of Thrace, Greece 5 Department of Obstetrics & Gynecology, Democritus University of Thrace, Greece Background: Survivin is a member of the inhibitor of apoptosis (IAP) family and is also involved in the regulation of cell division. Our objective was to study survivin expression in endometrial cancer and its correlation to clinicopathological parameters. Materials and Methods. A series of 110 cases of primary untreated endometrial carcinoma hosts were studied. Survivin immunoreactivity was assessed by immunohistochemistry using a polyclonal antibody, and evaluated semiquantitatively according to the percentage of cells demonstrating distinct nuclear or diffuse cytoplasmic staining. Correlation was made with tumor stage, grade, myometrial invasion, and histologic type. Association with disease outcome was also investigated. Results. Survivin overexpression, indicated by nuclear and cytoplasmic staining of at least 10% of the tumor cells, was found in 47 (42.7%) cases. A statistically significant association was demonstrated between survivin overexpression and FIGO stage (p=0.048), histological grade (p=0.024) and myometrial invasion (p=0.021). No significant association was found with histologic type of the tumor (p=0.164). Interestingly, patients with survivin positive tumors had significantly shorter survival rate (p=0.048). Conclusions. Survivin is a specific marker of endometrial cancer and strongly correlates with tumor stage, grade and myometrial invasion. It is also an independent prognostic indicator of poor outcome.

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PP3-15 PROGNOSTIC SIGNIFICANCE OF CYTOPLASMIC EXPRESSION OF C-erb-B2 IN ENDOMETRIAL CARCINOMAS Maria Lambropoulou1, Dimitrios Stefanou2, George Alexiadis1, Demetrio Tamiolakis1, Grigorios Tripsianis3, Ekaterini Chatzaki4, Gerasimos Vandoros1, Anastasia Kiziridou1, Nikolas Papadopoulos1 1 Department of Histology-Embryology, Democritus University of Thrace, Greece 2 Department of Pathology, University of Ioannina, Greece 3 Department of Medical Statistics, Democritus University of Thrace, Greece 4 Democritus Department of Pharmacology, University of Thrace, Greece Overexpression of the epidermal growth factor type II receptor cerb-B2 has been associated with resistance to chemotherapy and poor survival in several human tumors. The aim of this study was to investigate the expression of cerbB2 in endometrial cancer with attention both to membranous and cytoplasmic staining, to study its correlation to established clinicopathological parameters, and to elucidate the significance of cytoplasmic signaling. Tumor tissue from 110 patients with primary untreated endometrial carcinomas was available. C-erb-B2 reactivity was assessed by immunohistochemistry using a polyclonal antibody, and evaluated semiquantitatively according to the percentage of cells demonstrating membranous or diffuse cytoplasmic staining. Correlation was made with tumor stage, grade, myometrial invasion, and histologic type. Association with disease outcome was also investigated. C-erb-B2 overexpression, indicated by membranous and cytoplasmic staining of at least 10% of the tumor cells, was found in 47 (42.7%) cases. Cytoplasmic expression of c-erb-B2 was observed more frequently than membranous (69.09% versus 5.5%), and was correlated to established clinicopathological parameters. Synchronous cytoplasmic and membranous signaling was noticed in 7.9% of cases. A statistically significant association was demonstrated between c-erb-B2 overexpression and FIGO stage (p=0.048), histological grade (p=0.024) and myometrial invasion (p=0.021). No significant association was found with histologic type of the tumor (p=0.164). Interestingly, patients with c-erb-B2 positive tumors had significantly shorter survival rate (p=0.048). These results indicate that c-erb-B2 is a specific marker of endometrial cancer and strongly correlates with tumor stage, grade and myometrial invasion. It is also an independent prognostic indicator of poor outcome. C-erb-B2 antibody demonstrated a similar cytoplasmic and membranous staining. Cytoplasmic staining is as important as membranous staining and is a specific finding as well. PP3-16 IMMUNOHISTOCHEMICAL EXPRESSION AND CORELATION OF PROLIFERATING CELL NUCLEAR ANTIGEN (PCNA) AND MIB1 IN HYPERPLASTIC AND NEOPLASTIC ENDOMETRIUM: POSSIBLE PROGNOSTIC VALUE Ermina Iljazovic1, Elmir ýiþkusic1, Zinaida Karasalihovic1, Jasminka Mustedanagic2, D. Ljuca, Silvija Avdic1, Jugoslav Stahov3 1 University Clinical Center Tuzla, Polyclinic for Laboratory Diagnostic, Bosnia and Herzogovinia 2 University Clinical Center, Clinic for Gynecology and Obstetricion, Bosnia and Herzogovinia 3 Statistic Department, Nature Mathematic science, University of Tuzla, Bosnia and Herzogovinia Background: Endometrial neoplasia describes a spectrum of proliferative disease that constitues both a morphologic and biologic continuum beginning with mild hyperplasia and ending with poorly differentiated adenocarcinoma. The proliferative

activity of a hyperplasia and tumor has long been considered to bear a relationship to its clinical course. The main goal is to examine immunohistochemical expression of proliferation indices (PCNA and MIB1) in hyperplastic endometrium and endometrial adenocarcinoma and to identify possible prognostic value one of them. Methods: We conducted a retrospective study of 60 patients who had simple (n=20), complex (n=20) endometrial hyperplasia with or without atypia and endometrioid adenocarcinoma (n=20). The archival paraffin blocks from the uterine specimens were recut and assessed for histologic reexamination and PCNA and MIB1 immunostaining. Results were statistically analyzed with respect to marker expression and localization to the stromal or glandular component. Results: The PCNA index in the all forms of hyperplasia and adenocarcinoma was higher than MIB1 index (p< 0,0001; CI: 95,1%, median difference 4,5) in both, stroma and glandular components. There were no significant differences (one-factor analysis of variance [ANOVA]) in PCNA expression in hyperplastic endometrium and endometrial adenocarcinoma (F=0.48; P= 0 62; df =2,57). The MIB1 score was significantly lower and more heterogeneous in different hyperplastic and neoplastic lesions. Statistically significant difference (Scheffe test) was found in MIB1 immunoreactivity between simplex and complex hyperplasia (p= 0,0006), and complex hyperplasia and adenocarcinoma (p<0,0001), while there is no difference between simplex hyperplasia and adenocarcinoma (p=0.84). Conclusion: PCNA has a higher but constant expression in endometrial carcinoma and in hyperplasia than MIB1. These findings support that MIB1 may play a role in endometrial carcinogenesis, and that MIB1 index seems to be a better and significant prognostic parameter in hyperplastic endometrium than PCNA. PP3-17 A STUDY COMPARING EFFECTS OF INTRAUTERINE LEVONORGESTREL AND SYSTEMIC MEDROXYPROGESTERONE AS TREATMENT FOR ENDOMETRIAL HYPERPLASIA. ASSESSMENT BY LIGHT MICROSCOPY AND MORPHOMETRY Anne Oerbo1, Anne Beate Vereide2, Marit Arnes3, Kurt Larsen1, Inger Pettersen1, Bjorn Straume4 1 Department of Pathology, University Hospital of Northern Norway, N-9038 Tromsø, Norway 2 Department of Obstetrics and Gynecology, University Hospital of Tromsø, N-9038 Tromsø, Norway 3 Department of Pathology, Institute of Medical Biology, Faculty of, Norway 4 Institute of Community Medicine, Faculty of Medicine, University of Tromsø, N-9038 Tromsø, Norway Background: Endometrial carcinoma, showing increasing incidence in the Western world is generally developing through endometrial hyperplasia. Correct diagnostics and optimal treatment of endometrial hyperplasia is therefore of great importance. The female sex steroid hormone progesterone and different synthetical mimics, progestins, have shown a potent antiproliferative effect in the human endometrium. This effect has been utilized in clinical regimens in the treatment of endometrial proliferative disorders, endometrial hyperplasia as well as endometrial cancers. According to the literature no standard therapy regimen for progestine has been agreed upon in treatment of endometrial hyperplasia. To investigate if local application of the levonorgestrel impregnated intrauterine device was a better therapy for endometrial hyperplasia (EH) compared to per oral gestagen treatment, two patient groups were compared. The diagnostic evaluation was based on subjective (WHO criteria) and objective (prognostic data-based morphometric and stereological method/ D-score, predicting the risk of cancer development for each single patient) evaluation. Method: Women between 30 and 70 years with EH and D-score>0 were treated with levonorgestrel impregnated intrauterine device (n=26) and the results compared to a historic group of women treated with

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per oral gestagen (n=31). In both treatment groups only patients with low risk (D-score >1) and uncertain risk (D-score 0-1) of cancer development were included. Endometrial specimens were investigated prior to treatment and after 3 months of therapy. The endometrial samples from the two groups were examined by light microscopy and objective databased morphometry to assess tissue characteristics and to evaluate nuclear size variation. Results: After 3 months all patients treated with levonorgestrel intrauterine device showed regression of hyperplasia, whereas, 14 of 31 patients in the per oral group still had persisting disease. Morphometric analysis showed reduction in nuclear size for both treatment groups, including the D-score >1 as well as the D-score 0-1 patients. However, the reduction was most obvious for the levonorgestrel intrauterine device treated patients with initial Dscore of 0-1. Conclusion: The present study indicates that levonorgestrel intrauterine device is a superior alternative to per oral treatment of endometrial hyperplasia. This study showed that the hyperplasia patients with the highest malignant potential (Dscore 0-1) were those taking most benefit from local high dose levonorgestrel therapy. PP3-18 bcl-2, BAX AND APOPTOSIS IN ENDOMETRIAL HYPERPLASIA AFTER HIGH DOSE GESTAGEN THERAPY Anne Oerbo1, Anne Beate Vereide2, Turid Kaino3, Georg Sager4 1 Department of Pathology, Institute of Medical Biology, Faculty of, Norway 2 Department of Obstetrics and Gynecology, University Hospital of Tromsø, N-9038 Tromsø, Norway 3 Department of Pathology, University Hospital of Northern Norway, N-9038 Tromsø, Norway 4 Department of Pharmacology, Institute of Medical Biology, Faculty of, Norway Objectives. The aim of the study was to investigate apoptosis as a growth regulatory mechanism of gestagen in endometrial precancers and to compare differences in the immunohistochemical expression of proteins in the apoptotic cascade after high and low dose gestagen regimens. Method. Preand post treatment paraffin-embedded endometrial hyperplasia specimens from women treated with levonorgestrel intrauterine device (n = 26) and women treated with 10 mg medroxyprogesterone for 10 days per cycle (n = 31) were examined for changes in the expression of Bcl-2 and BAX and the amount of apoptosis after 3 months of treatment. Expression in tissue specimens for Bcl-2 and BAX were evaluated by Hscore. Apoptosis was assessed by counting apoptotic cells per 100 negative cells within ten different high power field (40X). Results. All the patients in the IUD group (n=31) but only about half of the patients in per oral group (16 of 26) responded to treatment. The glandular reduction in Bcl-2 expression was markedly greater for the IUD patients than for the patients who received oral gestagen. The decrease in BAX expression after IUD treatment was less than the reduction of Bcl-2. Decrease in glandular Bcl-2 after 3 months of treatment was coincident with a significant increase in the measurable amount of apoptotic bodies. In stromal cells increase in expression of Bcl-2 and BAX was found after gestagen treatment, the response being more extensive for the IUD group. The non- responders had no Bcl-2 expression in stroma after 3 months of therapy. The number of apoptotic bodies in endometrial glands increased significantly after 3 months of treatment with gestagen intrauterine device. No difference was observed for the per oral therapy. Conclusion. Our results indicate that proteins in the apoptotic cascade are regulated by gestagen therapy in human endometrial precancers. Expression of these proteins is shown to be dependent on type and administration form of the gestagen. Stromal Bcl-2 expression appears to be a potential biomarker which can separate responders of gestagen treatment from non-responders after oral administration.

PP3-19 P16 INK4A IMMUNOSTAINING: A USEFUL TOOL TO IDENTIFY OCCULT PREMALIGNANT LESIONS OF THE UTERINE CERVIX IN HPV-POSITIVE PATIENTS Susana Moyano1, Stefania Landolfi1, Sonia Garcia2, Natalia Basini1, Immaculada Alonso2, Elias Campo1, Aureli Torné2, Jaume Ordi1 1 Department of Pathology, Hospital Clinic, Barcelona, Spain 2 Department of Obstetrics and Gynecology, Hospital Clinic, Barcelona, Spain Background: The usefulness of p16INK4a in reducing interobserver variability; its ability to detect small foci of cervical intraepithelial neoplasia (CIN); and the close relationship between high-risk human papillomavirus (HR-HPV) infection and CIN, led us to evaluate whether p16INK4a staining could help to recognize under- or over-estimated CIN lesions, which may have clinical impact in the management of cervical disease. Method: Of 1259 women having a histological study and a simultaneous HR-HPV detection using the Hybrid Capture 2 (HC2) test, we selected all patients (n=139) testing positive for HR-HPV having a negative biopsy (group A), 26 women testing negative for HR-HPV with a biopsy of CIN 1 (group B) and 11 women testing negative for HR-HPV and having a biopsy of CIN 2-3 (group C). The following groups were randomly selected as controls: 50 cases negative for HR-HPV and with negative biopsy (group D); 50 cases positive for HR-HPV and with biopsy of CIN1 (group E) and 50 cases positive for HR-HPV and with biopsy of CIN2-3 (group F). Results: All cases from group D were either completely negative or showed only focal p16INK4a immunostaining, whereas all biopsies from group F were positive for p16, with 98% showing diffuse and strong staining. Thirtyfour out of 139 biopsies (24.5%) from the group A were positive for p16INK4a. Thirty of these cases (21.6%) were reclassified as harboring a CIN (11 CIN 1, 19 CIN 2/3) after reevaluation. Both the number of cases reclassified as CIN of any grade or as CIN 2/3 were significantly higher for cases with HR-HPV load above 100 RLU (p<0.005). Conclusion: Our results indicate that particular attention should be paid to biopsies from patients having positive HC2. The risk of harboring small foci of CIN of any type or a CIN 2/3 is significantly higher for patients with high HR-HPV load. Immunostaining with p16INK4a should be considered as a highly desirable adjunct to the histologic evaluation of cervical biopsy specimens. PP3-20 ATYPICAL (BIZARRE) STROMAL CELLS IN VARIOUS CONDITIONS OF THE FEMALE GENITAL TRACT: AN IMMUNOPATHOLOGIC REPORT OF 9 CASES Stolnicu Simona1, Gonzalez-Rocha Talina2, Mendoza Eladio3, Nogales Francisco F2 1 University Of Medicine Targu Mures, Romania 2 University Of Granada, Spain 3 Hospital Virgen Del Rocio, Seville, Spain INTRODUCTION: Atypical stromal cells of the female genital tract have been reported in various conditions of the vulva, vagina, cervix and endometrium, predominantly associated with polypoid lesions. Their atypicality includes them in the differential diagnosis of various conditions. Their origin and pathogenesis are unknown. MATERIAL AND RESULTS: We describe the presence of atypical stromal cells in a series of 8 cases comprising: fibroepithelial polyp of the vulva (1), fibroepithelial polyps of the vagina (3), normal cervix (1), of stromal response to a squamous carcinoma of the cervix (1 case) and endometrial polyps (2). Immunohistochemical studies were performed in order to elucidate its origin and proliferative capacity using the following antibodies:CAM 5.2, desmin, S 100 protein, actin, caldesmon, RE, RP and Ki 67. CONCLUSIONS: These studies demonstrated fibroblastic or myofibroblastic features, suggesting that they may represent a reactive or degenerative process of mesenchymal stem cells from

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subepithelial stroma. Although they have a benign clinical course, awareness of the pathologic and immunohistochemical spectrum of these cells is crucial in their correct differentiation from other lesions as microinvasive carcinomas, sarcomas, blue nevi and intermediate trophoblastic lesions, which also exhibit bizarre stellate cells. The differentiation with superficial myofibroblastoma is difficult since this entity does also share this cellular component that originates from a specialized band of subepithelial mesenchyme in the lower female genital tract PP3-21 FOLLICULAR-LIKE PATTERN IN METASTASES OF FEMALE GENITAL TRACT LEIOMYOSARCOMAS AND RHABDOMYOSARCOMAS. A PITFALL IN DIFERENTIAL DIAGNOSIS Simona Stolnicu1, Talina Gonzalez-Rocha2, Jose Fernandez Aneiros3, Luis Jose Sarasa3, Lucian Puscasiu1, Francisco F Nogales4 1 University Of Medicine Targu Mures Romania 2 University Of Granada Spain 3 Fundacion Jimenez Diaz Madrid Spain 4 University Of Medicine Granada Spain INTRODUCTION: We present an unusual and unreported follicular-like change in the abdominal and vaginal metastases of four cases of leiomyosarcoma (LMS) and rhabdomyosarcoma (RMS) of the female genital tract; three of them originated in the uterus and the remaining one in the vaginal wall. RESULTS: This pattern was not present in the primary tumour and consisted in a moderately atypical spindle cell population, sometimes exhibiting lobular growth, which presented many pseudofollicular spaces with an apparent cuboidal lining and basophilic contents in their lumina. Primary tumours corresponded to 3 highly differentiated leiomyosarcomas of the uterus and vagina respectively and a carcinosarcoma with rhabdomyosarcoma component in the remaining one. Immunohistochemically, the tumour cells from the primary and metastases were diffusely positive for vimentin, actin, desmin, caldesmon and myosin in the last case, which demonstrated the smooth or striated muscle identity of the tumours. The luminal contents were positive for both mucicarmine and PAS stains. CONCLUSIONS: Since similar follicular- like spaces are characteristic of tumours such as juvenile granulosa cell tumour (JGCT )and small cell carcinoma associated with hypercalcemia (SCC with HC), this phenomenon should be taken into account in their differential diagnosis which should also include the alveolar patterns of various sarcomas as well as malignant melanoma which also may conform such pseudofollicular spaces. PP3-22 REPRODUCIBILITY OF CERVICAL HISTOLOGIC INTERPRETATIONS BETWEEN PATHOLOGISTS Alenka Repše Fokter, Simona Šramek Zatler, Boris Kavþiþ, Zlatko Iterniþka, Marjeta Jene Kladnik Department of Pathology and Cytology, Celje General Hospital, Celje, Slovenia Background: The interpretive reproducibility of cervical cytology and histopathology is critical to cervical cancer prevention programs. In the medical community, histopathologic interpretations are generally considered as the reference standard upon which treatment of cervical disease is based. Therefore the aim of our study was to determine the interpathologist and intrapathologist reproducibility of histologic cervical specimen interpretations at our Department. Methods: We reviewed 60 randomly selected histologic cervical specimens, which were originally diagnosed by one of four pathologists at our Department. The original number of specimens was equally distributed between the pathologists (each pathologist 15 specimens). After review we calculated intraobserver reproducibility for each pathologist (15/15) and interobserver reproducibility, where all the pathologists had to interpret all 60

histologic specimens. We also compared cytologic and histologic interpretations if cytologic interpretations were available. Results: The average interobserver agreement was 73,8% (70% - 78,3%). The lack of reproducibility was most evident for negative and less severe interpretations. For high grade SIL lesions the average reproducibility was 91,7% (88,9%-97,2%). The average intraobserver agreement was 76,7% (60%-86,7%), and for high grade lesions 93,8%. Cyto-histologic correlation showed excellent results (100%) for H-SIL lesions and moderate for low grade lesions. Conclusion: Interpretive variability is substantial in cytology and histopathology. Our results showed excellent reproducibility of cervical histologic specimens for high grade squamous lesions and only moderate for low grade squamous lesions, which is in concordance with some previous studies on larger series. PP3-23 OVARIAN CELLULAR FIBROMAS: A CLINICOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL ANALYSIS OF TEN CASES Neli Basheska, Irina Prodanova, Katerina Kubelka-Sabit, George Zografski Department of Histopathology and Clinical Cytology, Institute of Radiotherapy and Oncology, Medical Faculty, Skopje, Republic of Macedonia BACKGROUND: Traditionally, cellular fibroblastic tumors of the ovary were classified as either cellular fibroma (CF) or fibrosarcoma. A recent study suggests that cellular fibromatous neoplasms with bland cytology and elevated mitotic counts are associated with favourable prognosis and should be diagnosed as "mitotically active cellular fibroma" (MACF) rather than fibrosarcoma. In addition to clinicopathological features, immunohistochemistry may aid in further differentiating between CF and MACF, but its role has not been analyzed so far. METHOD: We retrospectively analyzed the clinicopathological and immunohistochemical features of 10 cases of ovarian cellular fibroblastic tumors diagnosed either as CF or fibrosarcoma in the last seven years. Patient records and archival pathology specimens were reviewed and immunohistochemistry was performed using pan-cytokeratin, EMA, vimentin, inhibin-alpha, calretinin, CD10, CD99, alpha-smooth muscle actin (SMA), desmin, S-100, c-kit, estrogen (ER), progesterone receptor (PR), p53, bcl-2, and MIB-1 antibody. RESULTS: Utilizing criteria proposed by Irving et al. the tumors were reclassified as CF (0-3 MFs/10 HPFs, n=5) and MACF (>4 MFs/10 HPFs, n=5). The mean age of patients with CF and MACF was 44 and 36 years, respectively. All tumors were unilateral, and the mean tumor size of CFs was 6.0 cm and 13.3 cm for MACFs. The majority of the tumors were solid; four of them had a cystic component, while ovarian surface rupture was present in one CF and one MACF. All tumors consisted of cellular, intersecting bundles of spindle cells showing slight or moderate pleomorphism. The mean highest mitotic count was 2.3 MFs/10 HPFs for CF, and 7.6 MFs/10 HPFs for MACFs. Follow-up of 4 to 79 months (mean 38 months) was available in 9 patients and was uneventful in all cases. One patient with MACF died 15 days following the operation as a result of the intercurrent disease. Immunohistochemical analyses showed that spindle cells in majority of the tumors were immunoreactive for vimentin, alphaSMA, inhibin-alpha, calretinin, PR, and bcl-2. All tumors were negative for pan-cytokeratin, EMA, CD10, CD99, c-kit, ER, and p53, while one CF was positive for S-100, and one MACF showed positivity for desmin. In addition, the MIB-1 labeling index (LI) in MACFs was higher (mean 14.4%, range 10-25%), than that in CF (mean 5.6%, range 3-10%). CONCLUSION: Our results confirm the immunophenotypic similarity between ovarian fibromas and cellular fibromatous neoplasms, and suggest that the use of MIB-1 LI may help in differentiating between CF and MAFC.

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PP3-24 PRIMARY MELANOMA OF THE UTERINE CERVIX – CASE REPORT Grazyna Pasz – Walczak Department of Pathology, Chair of Oncology, Medical University of Lodz, Poland Only 5% of melanocytic malignancies in women originate in the genitalia. Primary malignant melanoma of the cervix is an extremely rare histological variety of cervical tumor, with only 54 cases reported. These lesions arise from melanocytes, which may be found within the mucosa lining the female genital tract. Primary malignant melanoma of the cervix is diagnosed by the presence of junctional melanocytic abnormality and the absence of distant metastases. We present a case of the malignant melanoma of the uterine cervix in 84-year-old postmenopausal women. The suspition of malignant melnoma was recognaised on a routine Papanicolaou-stained cervical smear. The gynecologic examination showed a cervical dark tumor, with extention into the vagina, the parametrium was free. Microscopic examination of the biopsy led to diagnosis of malignant melanoma (Fig.1). The cells were positive for S-100 (Fig.2)and HMB45 (Fig.3) in immunohistoceminal stain.The tumor was stage IIA of the International Federation of Gynecology and Obstetrics classification. Chest x-ray and abdominopelvic computed tomografy scanning were normal. Radical hysterectomy with bilateral salpingo-oophorectomy, pelvic and paraaortic lymphadenectomy, partial vaginectomy, and adjuvant radiation or chemotherapy have been advocated by some investigators. In our case, due to old age the patient received palliative radiation therapy to the pelvis, total dose 20Gy. Cervical melanoma is a rare disease of which no retrospective studies exist, only case reports.Primary malignant melanoma of the uterine cervix carries a very poor prognosis. The small number of reported cases makes it difficult to evaluate the diagnosis and treatment. Immunohistochemistry plays very important role in the diagnosis of this tumor in the early stage. This is essential since cervical melanoma is incurable with standard therapies. PP3-25 THE IMPORTANCE OF USING THE CYTOBLOCK TECHNIQUE FOR WOMEN WITH BLEEDING SYMPTOMS Mathilde E. Boon1, Saskia M. Duineveld2, Elisabeth Ouwerkerk-Noordam1, Dwen Dias2 1 Leiden Cytology and Pathology Laboratory, Leiden, The Netherlands 2 Goa Medical College, Department of Pathology, Goa, India Background: Cervical sampling was changed fundamentally when in the eighties of the past century the Cytobrush was introduced. Finally, the Ayre spatula was completely replaced by various brushes. The more immature lesions, including the preneoplastic CINs, are often located in the endocervical canal and are effectively sampled by the brushes. Our results indicate that inflammatory changes can lead to cervical bleeding, probably because inflammation is accompanied by angiogenesis. It is a well known fact that (pre)invasive cervical carcinoma is rich in newly formed small blood vessels, leading to post-coital bleeding. Our purpose was to use such cervical samples to prepare Cytoblocks and exploit the paraffin section made thereof for additional immunostaining for women with bleeding symptoms. Methods: The 261 cervical samples were processed by the Shandon Cytoblock Preparation System. The vials still containing the brush were placed in a shaker for 10 minutes to dislodge the material trapped between the bristles of the CervexBrush. Specimens fixed in BoonFix were concentrated by centrifugation (10 min at 3000 rpm). The Cytoblock cassette (containing the concentrated material infiltrated with agar) was removed from the Cytospin and histoprocessed in the Pathos (Milestone, Italy) applying m sections were cut and stainedPmicrowave technology. From each block several 4 with

the Papanicolaou method. On the Papanicolaou-stained paraffin sections made thereof a histological diagnosis was given rendered on the minibiopsies. Results: Of the total 261 women screened, 237 had a benign diagnosis, with only 138 having a completely normal histology. In 42 cases an inflammation was suggested. The CIN cases contained minibiopsies with classical feature of cervical neoplasia which were discerned with ease in the excellent paraffin sections with optimal nuclear morphology. In the two cases of squamous cell carcinoma, many minibiopsies examined displayed squamous cells connected with extended desmosomes and hyperchromatic nuclei containing a macronucleolus. The (pre)invasive cases had many Ki-67 positive nuclei displaying an S-phase staining pattern. In the Ki67 stained sections, the glandular architecture of the two AIS cases and the two adenocarcinoma cases was highlighted. Conclusion: In the histologic paraffin sections there were enough minibiopsies to allow concise diagnosis including evaluation of proliferation. Signs of cervical angiogenesis, including post-coital bleeding, can be a strong argument to prepare Cytoblocks from samples collected by sampling brushes. PP3-26 FREQUENCY AND CHARACTERIZATION OF ANTERIOR PITUITARY COMPONENT IN OVARIAN MATURE CYSTIC TERATOMAS Elena Rigoli, Donata Micello, Nikolaos Papanikolau, Stefano La Rosa, Cristina Riva, Carlo Capella Department Of Human Morphology, University Of Insubria Varese, Italy Background: Adenohypophyseal tissues and pituitary-type tumors, namely functioning corticotroph and lactotroph adenomas, have been rarely reported in ovarian mature cystic teratomas (OMCTs), but the frequency of this feature is, to date, unknown. The purpose of this study was to evaluate the incidence of a pituitary component in a large series of OMCTs. Methods: An archival series of 200 consecutively collected and of 4 selected OMCTs was revised. On the basis of the morphological finding of cellular nests or solid islands, suggesting an endocrine tissue, 56 cases were selected for the immunohistochemical study. Immunoreactions for Chromogranin A, GH, PRL, ACTH, ȕFSH, ȕLH, ȕTSH, Į-subunit and thyreoglobulin were routinely performed. Results:Anterior pituitary cells were detected in 8 OMCTs including 6 cases among the consecutive series (incidence: 3%) and 2 selected cases. Anterior pituitary component was frequently observed near a struma ovarii (6 cases) in form of small groups or strands of polygonal endocrine cells encircling or penetrating into the thyroid parenchyma. In one case, an adenomatous micronodule associated with a strumal carcinoid was present . In two cases the pituitary cells were localized beneath a respiratory mucosa. The following immunoreactivities were obtained: Chromogranin A (7/8 cases), PRL (6/8 cases), GH (5/8 cases), Į-subunit (4/8 cases), ȕFSH (2/8 cases), ACTH (1/8 cases). The adenomatous micronodule showed diffuse PRL+ e focal GH+. Conclusions: Adenohpophyseal cells are present in 3% of OMTCs. The immunoprofile of pituitary cells demonstrates PRL expression in the majority of the cases, suggesting that hyperprolactinemia could contribute to the clinical syndrome of amenorrhea and/or infertility in a subset of young patients with OMTCs PP3-27 LYMPHOEPITHELIOMA-LIKE CARCINOMA OF THE ENDOMETRIUM: AN UNUSUAL VARIANT OF ENDOMETRIAL CARCINOMA Kalliopi Patsiaoura, Ioannis Amplianitis, Afroditi Pantzaki, Alexandra-Anastasia Moulla, Despina Alataki Hippocration General Hospital, Greece Lymphoepithelioma is a distinct neoplasm with unique histologic features and a consistent etiologic association with the EB virus. The neoplasm was first identified in the nasopharynx, but since

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then carcinoma identical to lymphoepithelioma and designated lymphoepithelioma-like has been reported in many other organs. While lymphoepithelioma-like carcinoma of the cervix is well established lymphoepithelioma-like carcinoma of the endometrium is extremely rare with 3 cases reported to date. A 73 year-old woman with postmenopausal bleeding was admitted to hospital. The physical examination and the laboratory evaluation were both unremarkable. A total hysterectomy with bilateral salpingo-oophorectomy was performed. The hysterectomy specimen revealed a tan polypoid tumor filling the endometrial cavity and protruding partially into the cervical canal. Microscopically, the myometrium was diffusely infiltrated by a neoplasm composed mainly of plump cells with distinct cytoplasmic borders, vesicular nuclei and prominent nucleoli. The neoplastic cells were arranged mostly in groups with a syncytial appearance surrounded by an intense inflammatory infiltrate mainly with lymphocytes and plasma cells. Immunohistochemically, the neoplastic cells revealed strong keratin and EMA as well as focal estrogen and progesterone receptors positivity. The immunostains for synaptophysine, chromogranine, a-SMA, CD10, inhibin, and CEA were totally negative. In accordance with the previously reported cases the immunohistochemical analysis for the EBV was negative. The morphologic and immunohistochemical findings supported the diagnosis of a lymphoepithelioma-like carcinoma of the endometrium. We present this case as being both interesting and extremely rare and review the related literature. PP3-28 UTERINE LIPO-LEIOMYOMA WITH FOCI OF CARTILAGINOUS DIFFERENTIATION Kalliopi Patsiaoura, Ioannis Amplianitis, Aphroditi Pantzaki, Eleftherios Anagnostou, Anastasios Chatzitolios Department of Pathology, Hippokrateion General Hospital, Thessaloniki, Greece The presence of foci with cartilaginous differentiation within an otherwise typical uterine leiomyoma or lipoleiomyoma is an extremely rare phenomenon, with only 3 reported cases so far in the literature. A 47-year old woman with a history of HGSIL, submitted to hospital due to uncontrolled vaginal bleeding. A total abdominal hysterectomy with bilateral salpingooophorectomy was performed. In the uterine corpus two intamural tumors, with diameters of 5,5 cm and 3 cm respectively, were observed. The tumors were both wellcircumscribed, whitish with a soft consistency. Microscopically, the tumors were mostly composed of intervening fascicles of smooth muscle cells. Increased cellularity, cytologic atypia, necrosis or mitotic activity were totally absent. Between the smooth muscle cells, foci composed of mature cartilaginous tissue, as well as areas composed of well-differentiated adipocytes were noticed. These areas showed strong positivity for the S-100 immunostain. The histologic and immunohistochemical findings were consistent with the diagnosis of uterine lipoleiomyoma with foci of cartilaginous differentiation. We present the case because of its rarity and we review the few reported cases in the literature, regarding its histogenesis and the related differential diagnostic problems with the other mesenchymal neoplasms of the myometrium. PP3-29 ENDOMETRIOSIS AND ENDOMETRIOID CARCINOMA OF THE OVARY Laura Yébenes1, Paloma Ramos1, Ana Blasco1, Pedro Valenzuela2, Isabel Álvarez3, Antonio Ruíz1 1 Pathology Department Príncipe de Asturias Hospital, Spain 2 Obstetrics and Gynaecology Department Príncipe de Asturias Hospital, Spain 3 Ramón y Cajal Hospital Pathology Department, Spain

BACKGROUND: We present a retrospective analisis of 22 cases of endometrioid ovarian carcinoma, reviewed to identify endometriosis and its malignant transformation. METHOD: Twenty-two patients with endometrioid ovarian cancer were included in the review. Their clinical and histological data were retrospectively reviewed. The origin of the tumor was considered endometriosis-related when the presence of malignant changes in endometriosis glands leading to endometrioid carcinoma were found. RESULTS: Endometriosis was detected in three cases (3/22=14%). One of them presented a clearly benign to malignant transformation area. In another patient, the transition zone was abrupt and present in both ovaries. In the third, a pre-menopausal woman, ovarian endometriosis with only focal endometrioid carcinoma was observed. The three of them had a clear-cell carcinoma component. The presence of a clear-cell component was significantly greater in patients with endometriosis than in patients without endometriosis. Each patient had a different clinical presentation: increase in abdominal perimeter, postmenopausal vaginal haemorrhage and hypermenorrhea. Preoperative CA 125 levels were avalaible in 15 of the patients (15/22=68%). Endometriosis was found in two of these 15 patients, both with the highest CA 125 measured levels, exceeding 1700 U/ml. In the remaining of the patients, including the third one with endometriosis, CA 125 value did not exceed 35 U/ml. CONCLUSION: Although this association is not very frequent, patients with ovarian endometriosis and high CA 125 serum level should be managed with special care, regardless of their pre-menopausal or post-menopausal status. PP3-30 PRIMARY OVARIAN HEMANGIOENDOTHELIAL SARCOMA - REPORT OF A CASE Ioannis Amplianitis Department of Pathology ‘‘Hippokrateion’’ General Hospital, Thessaloniki, Greece A primary ovarian hemangioendothelial sarcoma, also known as ovarian hemangiosarcoma or angiosarcoma is a very rare neoplasm - fewer than 25 cases have been recorded to date. A 89 year - old woman was admitted to the hospital because of a cystic tumor mass on her left ovary. She was offered a unilateral salpingo- oophorectomy. Grossly, the ovary revealed a unilocular cystic tumor of a maximum diameter 5.5 cm, with clear fluid contend, which histologically proved to be a benign serous cystadenoma. Peripherally and within the ovarian stroma, an infiltrating neoplasm composed of vascular spaces of varying size, lined by endothelial cells with prominent atypical appearance was recognized. Extra-ovarian expansion was not observed. Positivity for the vascular markers CD34 and Factor VIII confirmed the endothelial origin of the neoplasm and supported the diagnosis of ovarian hemangioendothelial sarcoma. The immunostains for pankeratin, inhibin and calretinin were negative. The neoplasm, though rare, raises differential diagnostic considerations, involving the germ cell tumors, mostly the immature teratoma, the ovarian lymphangiosarcoma and hemangiopericytoma, as well as the metastatic spread to the ovary from primacies in the spleen or the breast. The histogenesis of the neoplasm remains uncertain. The concurrence of an epithelial neoplasm, as observed in our case, has been previously reported. We present the case as being both rare and interesting and review the related literature, regarding the differential diagnostic problems and the histogenetic considerations. PP3-31 A CASE OF MUCINOUS MICROGLANDULAR ADENOCARCINOMA OF THE ENDOMETRIUM Hannachi Sassi Samia, Driss Maha, Mrad Karima, Abbes Imen, Dhouib Rym, Ben Hamida Naziha, Ben Romdhane Khaled Department of Pathology Salah Azaeiz Institute, Tunisia

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Microglandular adenocarcinoma (MGA) is a rare type of endometrium carcinoma with only thirteen cases have been reported in the literature, according to our knowledge. MGA may mimic morphologically some benign and malignant lesions of the cervix. We report the fourteenth case of a 63-year-old, para 7 gravida 7, postmenopausal Tunisian woman consulted for uterine bleeding of 3-month duration. Histopathologic evaluation of the curettage specimen made the diagnosis of mucinous MGA. The patient underwent a total hysterectomy and bilateral salpingooophorectomy with pelvic lymph node sampling. Histology showed a stage IB grade II mucinous MGA. Because MGA is an uncommon type of endometrium adenocarcinomas and has potential problems in differential diagnosis with some lesions of the cervix, we discuss the clinical, histological and immunohistochemical characteristics of MGA useful in differential diagnosis. PP3-32 ENDOCRINE AND IMMUNE PARALLELS IN LICHEN SCLEROSUS (LS) AND VULVAR INTRAEPITHELIAL NEOPLASIA (VIN): IMMUNOMORPHOLOGICAL STUDY Gleb Sychugov, Eugene Kazachkov, Ella Kazachkova, Eugenia Malachova Medical Academy, Chelyabinsk, Russia Background. Immunomorphological study of vulvar biopsy specimens taken of 19 women aged from 37 to 65 diagnosed with LS and VIN, grade 3 was undertaken (LS - 10, VIN - 9). Method. Paraffin sections were incubated with monoclonal antibodies to progesterone (PGR), estrogene (ER) receptors, CD3, CD4, CD20, CD68, HPV types 6,11,18, polyclonal antibodies to IgA, IgG, IgM (“Novocastra”). Results. HPV was found in cellular nuclei of stratified squamous epithelium (SSE) in all subjects. Volume density of ER-positive cells in LS was 1.65±0.14%, in VIN 2.95±0.19%, while PGR-positive - 3.93±0.21% and 5.46±0.23% respectively. ER-positive and PGR-positive cells expression in SSE was mainly noted in basal layers, particularly in acanthotytic bands, where hormone sensitive cells representation was total. Volume density of various subpopulations of lymphocytes and macrophages did not change significantly aggravated: for CD4lymphocytes it averaged 0.5±0.08%, CD8-positive – 1.05±0.16%. CD4/CD8-ratio was 0.47. The number of IgGproducing cells remained stable and averaged 39%. As VIN got more severe the number of IgA-producing cells decreased from 28% in LS to 17% in VIN3, the number of IgM-producing cells increased from 33% to 44%. Conclusion. Thus, as VIN becomes more severe some changes in local humoral immune status are observed, particularly an increased level of IgM-producing cells. Cellular immunity transformation is expressed in the decreased number of CD4-lymphocytes.The number of cells expressing receptors to sex hormones increased significantly at the site of SSE growth. PP3-33 FIBROSIS AND SMOOTH MUSCLE METAPLASIA IN INTESTINAL ENDOMETRIOSIS Makoto Jinushi1, Atsushi Arakawa2, Keiko Abe2, Toshiharu Matsumoto2, Tomoko Itoga1, Mari Kitade1, Hiroyuki Takeuchi1 1 Department of Obstetrics and Gynecology, Japan 2 Department of Pathology, Japan Background: In case of endometriosis of the colon, serosa and muscularis propria are involved much more frequently than submucosa and mucosa. This finding suggest that lesions of endometriosis spread from the serosa to the mucosal layer. Itoga et al. examined the histological findings of endometriosis in the rectovaginal space, and showed that the degree of fibrosis and smooth muscle metaplasia might increase with relation of the age of ectopic endometrial tissue. This conclusion also suggested

that fibrosis and smooth muscle metaplasia could represent the chronic phase of ectopic endometrial lesion. We examined the degree of fibrosis in the ectopic endometrium in each layers of the colon, for decision of which lesions early or chronic phase are. Method: Three women with endometriosis of the rectum were archived from the medical records of our hospital. All of them underwent surgical resection of the rectum. These materials were fixed by 10% formalin at least overnight. Ninety-four lesions were selected from these three specimens. These lesions were located at the subserosa (ss-lesion) (13 lesions), at the muscularis propria (mp-lesion) (78 lesions) and at the submucosa (sm-lesion) (3 lesions). No lesions were observed in the mucosa (m-lesion). Each lesion was classified into four groups in accordance with the degree of fibrosis surrounding the endometrial duct. The classification of the degree of fibrosis was, Group1: endometrial duct without stromal tissue surrounding fibrosis, Group2: endometrial duct surrounded by only endometrial stroma, Group3: endometrial duct partially surrounded by fibrous tissue, Group4: endometrial duct completely surrounded by fibrous tissue. Results: 1) Ectopic endometrial tissues were most frequently observed in the muscularis propria (sm 3, mp 78 and ss 13). 2) The lesions in the muscularis propria showed various stromal and fibrosis features (Group1 17%, Group2 32%, Group 3 22% , Group4 29%). 3) On the other hand, the lesions in the submucosa and subserosa revealed some endometrial ducts surrounding by fibrosis, completely or focally. (sm: Group3 33%, Group4 67%; ss: Group3 30.8%, Group4 69.2%). 4) Especially, in the lesions of the subserosa, smooth muscle metaplasia was found in the extensive spread of fibrous tissue. Conclusion: Ectopic endometrium in the subserosa with prominent fibrosis and smooth muscle metaplasia might represent the long-standing lesion of the enodometriosis in the colon. The first site of endometrial tissue infiltrated into the colon could be identified as the subserosa of the colon. PP3-34 TISSUE MICROARRAY STUDY OF TUMOUR BIOMARKERS IN HPV INDUCED CERVICAL NEOPLASIA Ita Hadžisejdiü1, Maja Grahovac2, Dražen Kovaþ1, Maja Kraševiü1, Blaženka Grahovac1 1 Department of Pathology, Rijeka University School of Medicine, Croatia 2 Department of Dermatovenerology, Medical School, University of Zagreb, Croatia The aim of this study was to analyse expression patterns of tumour biomarkers Ki 67, p53, p16 (INK4A) and epidermal growth factor receptor (EGFR), in cervical carcinoma using tissue microarray technology (TMA). Methods: The expression of Ki67, p53, p16(INK4A) and EGFR was studied in series of 34 cervical cancer samples: 6 cervical intraepithelial neoplasia (CIN III), 14 squamous cell carcinomas (SCC), and 14 adenocarcinomas (ADC). TMA slides containing triplicate of 1.0 mm cores were constructed and subjected to immunohistochemical protein analysis. Results: High-risk HPVDNA was identified in 94 % (32/34) of the cases. HPV-16 predominated in SCC and CIN III, while HPV 18 and HPV 16/18 mixed infection were detected in ADC. Ninety-one percent of samples (31/34) exhibited strong proliferative activity (+2 and +3 Ki 67 immunopositivity), 64% (22/34) exhibited high expression of p16(INK4A), 73% (25/34) demonstrated high expression of p53 and 61 % (21/34) of uterine cervix neoplasia exhibited high EGFR expression. According to the histological type, SCC and ADC demonstrated high expression of Ki67, p16(INK4A) and p53. In CIN III only high expression was detected for Ki67, while p16(INK4A) and p53 were expressed at low level. Substantial differences were shown in EGFR expression: SCC and CIN III demonstrated high level of EGFR expression, in 92 % (13/14) and 100% (6/6) samples, respectively. In eighty-five percent of

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ADC’s the expression of EGFR was absent (9/14) or (+1) intensity (3/14). Only two tumours demonstrated moderate/strong (+2) level of expression. Conclusion: Our study has demonstrated the reliability of tissue microarray technology in protein expression studies. Preliminary data revealed substantial differences in tumour markers expression in different histological types of cervical neoplasia. To confirm these results, larger studies are needed. Especially, EGFR expression in ADC and adenosquamous carcinoma should be investigated to elucidate the role of EGFR expression in these histological types of cervical carcinomas. PP3-35 OVARIAN SEX CORD TUMOR WITH ANNULAR TUBULES - REPORT OF A CASE Milco Ristovski1, Rubens Jovanovic1, Vanja Filipovski1, Slavica Kostadinova Kunovska1, Elizabeta Milanova2 1 Institute of Pathology, Medical Faculty, Univesity "St. Cyril and Methodius" Skopje, R. Macedonia 2 Clinic for Gynecology and Obstetrics, Clinical Centre, Skopje, R. Macedonia The sex cord tumor with annular tubules (SCTAT) was discovered as a distinctive entity when its unusual appearing microscopic pattern was encountered repetitively in a pathology consultation practice. This tumor is a distinctive ovarian neoplasm, the prominent component of which shows morphological features intermediate between those of the granulosa cell tumour and those of the Sertoli cell tumour. The aim of this paper is to report the first case of SCTAT in our Institution. Methods: We used clinico-biochemical data, macroscopic and microscopic morphology analysis, as well as immunohistochemical analysis of FFPE tissue samples. Results: A 23-year-old woman presented hirsutism and menometrorrhagia. Tumorous formation on the left adnexa was palpated. Ultrasound findings indicated septate cystic tumor on the left ovary. Erythrocyte Sedimentation Rate was 30/60 and CA-125 more than 40 U. (ref. 30 U.). Karyotype of the leucocytes from the peripheral blood was 46, XX. Gross examination revealed that the tumor was unilateral with smooth surface. It measured 25x22x12 cm. Cut surface exhibited multilocular cystic compartments filled with translucent gelatinous content. The compartments were separated with yellowish septa of variable width. Microscopic examination showed simple and complex annular tubules with prominent basement membrane-like material around the tubules. The lumina of tubules contained hyaline bodies. Small sheets of solid clear cells were also present, as well as calcified necrotic fields. Tumor cells were positive for cytokeratin cocktail and alpha-inhibin, they showed intermediate signal for WT1 (Wilms Tumor 1) and negative stain for cytokeratin 5/6. We can conclude that SCTAT is a very rare entity with unclear histogenesis, unpredictable prognosis and, very often, with miss out on morphology recognition. PP3-36 BILATERAL OVARIAN WELL-DIFFERENTIATED SERTOLI-LEYDIG-CELL TUMOR(S) ASSOCIATED WITH UNILATERAL SEROUS CYSTADENOMA – A CASE REPORT Elvira Stacher, Farid Moinfar Department of Pathology, Medical University, Graz, Austria Background: Bilateral Sertoli-Leydig-Cell Tumors of the Ovary, especially in association with unilateral Serous Cystadenoma, are exceedingly rare. We present a case of a 61 year old woman with this diagnosis, in which the Sertoli-Leydig-Cell Component shows leiomyogenic (heterologous) differentiation. On routine examination bilateral ovarian cystic tumors were detected by ultrasonography. Bilateral oophorectomy was performed, the left ovary was sent for frozen section diagnosis. Macrosopically the

left ovary measured 27x16x7cm, had a weight of 1965 grams and showed multiple cysts with numerous greyish to yellowish solid nodules measuring up to 1cm. The right ovary (no frozen section diagnosis obtained) measured 4x2,5x2cm and and showed on cut surface a 2,5x1,5x0,5 cm brown to grey solid-cystic tumor. Method: The tissue was processed and stained (H&E, immunohistochemistry) according to standard protocols. To confirm the sex-cord stromal component, immunohistochemistry for Į-Inhibin (Serotec, dilution 1:20) was also performed. Results: Left ovary: The frozen section revealed a serous cystadenoma/adenofibroma. The examination of the formalinfixed paraffin-embedded tissue displayed in addition infiltrates of a well differentiated Sertoli-Leydig-Cell Tumor. The SertoliLeydig-Cell Tumor showed irregular infiltration of the cysts. The tumor cells lacked significant cytologic atypia and mitotic activity. Immunohistochemistry revealed intense and diffuse positivity for Į-Inhibin. Within the well-differentiated sex-cord stromal tumor there were mesenchymal areas showing fascicular arrangement of spindle cell with eosinophilic, fibrillary cytoplasm. These heterologous areas were immunoreactive for smooth muscle actin. Right ovary: The tumor of the right ovary resembled a well differentiated Sertoli-Leydig-Cell Tumor. Conclusion: To the best of our knowledge, this is the first described case of a Sertoli-Leydig-Cell Tumor of the ovary occurring in a serous cystadenoma. Leiomyogenic differentiation and bilaterality are further interesting features of this particular case. PP3-37 IS CD10 IMMUNEXPRESSION HELPFUL IN DETECTING OCCULT STROMAL CELLS IN OVARIAN ENDOMETRIOSIS? Funda Eren, Ipek Erbarut Marmara University School of Medicine Department of Pathology, Istanbul, Turkey The diagnosis of endometriosis is based on finding glands lined by endometrioid epithelium surrounded by densely packed small fusiform cells with scanty cytoplasm and bland cytology typical of nonneoplastic endometrial stromal cells. These features may be obscured by hemorrhage, inflammation and fibrosis and diagnosis of endometriosis could be difficult. CD10 has been shown to stain some non-hematological tissues including endometrial stromal cells. In this study our aim is to detect CD10 immunreactivity in cases of presumptive ovarian endometriosis and to determine the value of CD10 in achieving a definitive diagnosis. 14 cases of ovarian biopsies diagnosed as not diagnostic but compatible with endometriosis between the years 2001-2007, were retrieved from the archives of Marmara University Department of Pathology. All slides were reevaluated and selected sections were stained with CD10 by immunohistochemistry. A case of definitive endometriosis and an endometrial tissue were used as positive controls. 2 cases with follicular cysts and inclusion cysts were used as negative controls. In 12 of the 14 cases (86%) strong CD 10 immunreactivity was seen in the stroma underlying a mainly attenuated epithelium. These areas were unnoticed in H&E sections due to fibrosis or hemorrhage. Areas that may simulate endometriosis were negative for CD 10 both in the study cases and negative controls except one. In this one case luteinized follicular cells showed weak cytoplasmic positivity for CD10. In two cases (24%) CD10 immunreactivity were negative. Overall strong positivity for CD 10 is a specific marker for endometrial stromal cells but it should remain in mind that weak staining may be seen in luteinized cells. However, such lesions are not difficult to differentiate morphologically. In conclusion, our study showed that CD 10 immunreactivity is useful and can be used in differential diagnosis of controversial cases of endometriosis.

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PP3-38 HISTOPATHOLOGICAL CHANGES IN PREPUBERTAL RAT OVARIES SUBJECTED TO TORSION AND DETORSION: EFFECTS OF L-CARNITINE AND NACETYL CYSTEIN Ufuk Usta1, Mustafa Inan2, Hakan Erbas3, Nurettin Aydogdu4, Fulya Oz Puyan1, Semsi Altaner1 1 Trakya University, Medical Faculty, Department of Pathology 2 Trakya University, Medical Faculty, Department of Pediatric Surgery 3 Trakya University, Medical Faculty, Department of Biochemistry 4 Trakya University, Medical Faculty, Department of Physiology Background: We aimed to evaluate histopathological changes, to detect HIF-1Į levels and to determine MDA levels in prepubertal rat ovaries, which were subjected to torsion and detorsion and treated with L-Carnitine or N-Acetyl Cystein (NAC). Methods: Forty prepubertal female Sprague-Dawley rats were used in the study. In group 1, only laparatomy was performed. In group 2, right ovaries were subjected 3 h of ischemia and removed sugically. In group 3, an ischemic period of 3 h was followed by reperfusion for 24 h. In groups 4 and 5, either L-Carnitine (100mg/kg) or NAC (20mg/kg) was infused intraperitoneally on 150th min of ischemia and ovaries were reperfused on 180th min. After 24 hours of reperfusion right ovaries were removed. Blood samples were collected before sacrification of each rat to determine serum IL-6 levels. All ovarian tissues were subjected to histopathological examination and MDA levels were determined biochemically. Immunohistochemically HIF-1Į antibody was applied to all ovaries. Histopathologically a score between 0 and 9 was formed for each ovary, which was composed of sum of grades of vascular congestion, edema and paranchymal hemorrhage, each graded between 0 and 3. Immunohistochemically HIF-1Į staining was scored between 0 and 9 which was the sum of separate evaluations of staining intensities of granulosa cells, theca interna cells and theca externa cells, graded between 0 and 3. Results: HIF-1Į did not show any nuclear staining while there was expressive cytoplasmic staining with an unknown significance. Histopathological scores and tissue MDA levels as well as HIF-1Į scores, were significantly higher in groups 2 (all P<0.001) and 3 (P<0.01, P=0.05 and P<0.001 respectively) then group 4. They were higher significantly in group 2 (P<0.001) then group 5. Histopathological scores and MDA levels were significantly high in group 3 when compared to group 5 (P<0.01 and P<0.05 respectively. Serum IL-6 levels were significantly high in group 2 when compared to groups 4 and 5 (both P<0.01). Conclusion: Histological pathologies of ovarian tissues seem to decrease after 24 hours of reperfusion of the torsioned ovaries. Tissue MDA levels and serum IL-6 levels also support those results. However, reperfused ischemic ovaries treated by both L-Carnitine and NAC, showed better recovery than the untreated group, though LCarnitine seems to be more effective then NAC. PP3-39 PROGNOSIS OF SQUAMOUS CELL CARCINOMA (FIGO STAGE IA2, IB AND IIA) OF THE UTERINE CERVIX PREDICTED BY MALIGNANCY GRADING SCORE EVALUATION Tormod Eggen, Marit Arnes, Bjorn Moe, Straume Bjorn, Marika Ostman, Anne Orbo University Hospital of North Norway, Norway BACKGROUND Most reliable prognostic factor for squamous cell carcinoma (SCC) of the uterine cervix is staging according to FIGO. Pelvic lymph node metastasis implies less favorable prognosis and such patients are usually offered adjuvant treatment. Some patients lacking metastasis will also experience relapse and would probably benefit from adjuvant treatment, but predicting such tumor behaviour has proven difficult. The

malignancy grading score (MGS) system based on 8 histological parameters (structure, polymorphism, mitotic rate, mode and stage of invasion, vascular invasion, cellular response) was introduced in the seventies, but has not been taken into general use. We wanted to investigate if the MGS system and image analysis would be suitable tools for identification of high-risk metastasis-free patients. METHOD In the study 82 patients diagnosed with- and treated for early stage SCC at the University Hospital of North Norway were included. Nineteen patients had been given adjuvant treatment due to lymph node metastasis, lack of radical exciscion or high risk profile. Nine patients experienced relapse and 7 died during follow-up of 5-16 years. Information including lymph node metastasis, resection margins, depth of infiltration, tumor diameter and clinical stage according to FIGO was taken from the hospital files. On the biopsy material initially evaluated, an MGS-score was given in each case by two pathologists independently. Finally, image analysis was performed. RESULTS We found metastasis, unfree resection margins and deep infiltration to significantly predict a poor outcome. Interestingly, the MGS system proved highly significant in separating patients with high risk of relapse from those with low risk. This was true when the material was evaluated as a whole and also when node positive patients were excluded. The inter-observer reproducibility was evaluated by Cohen’s Kappa to 0,9. Image analysis proved unable to predict prognosis. CONCLUSION The MGS system is a fairly simple method and can easily be implemented into routine diagnostics. Its ability to predict relapse in patients lacking metastasis at time of initial surgery is therefore especially interesting. Image analysis offers no additional information. PP3-40 INVESTIGATION OF FASCIN PROGNOSTIC SIGNIFICANCE IN ENDOMETRIAL CARCINOMAS Mattheos Bobos1, Ioannis Kalogiannidis2, Alexios Papanikolaou2, Georgios Makedos2, Eleni Nenopoulou1 1 Department of Pathology, Aristotle University Medical School, Thessaloniki Greece 2 4th Department of Obstetrics and Gynecology, Aristotle University Medical School, Thessaloniki Greece Background: Fascin is an actin cross-linking protein that has been implicated in cell motility and migration. The absence or low expression of fascin in normal epithelia is dramatically altered in many human carcinomas. The aim of this retrospective study was to investigate fascin expression in endometrial carcinoma cases and correlate with clinicopathological factors and survival data. Methods: FFPE tissue specimens from 58 patients with endometrial adenocarcinoma were arrayed (5 cores of 0.6mm) using the tissue microarray technology. Clinical, pathological and survival data were collected from all patients and correlated with the immunohistochemical data. Results: The mean age of patients was 63 years (range 35-80). The majority of the cases (96%) was of endometrioid subtype and 79% of FIGO Stage I. Fascin expression was detected in 44/56 (76%) of cases, more common in grade 1 tumors, tumors with diameter <2cm, and with superficial myometrial infiltration (p = 0.2, p = 0.1 and p = 0.1, respectively). The estimate overall survival (OS), cancer related survival (CRS) and disease free survival (DFS) of patients with positive fascin were not significantly higher from those with lack of fascin expression. DFS was not significantly improved when expression was correlated with different clinicopathological parameters (age, FIGO staging, grade, myometrial infiltration, tumor diameter, extrauterine disease). Conclusion: According our data, it seems that fascin expression in patients with endometrial cancer is related with favorable grade, myometrial invasion and tumor diameter. In contrast to previous study we didn’t find any impact in patients’ survival.

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PP3-41 DOCUMENTATION OF 938 OVARIAN LESIONS (NEOPLASTIC-NONNEOPLASTIC) IN A PERIOD OF SIX YEARS Seyran Yigit, Betul Bolat Kucukzeybek, Mehmet Ali Uyaroglu Ataturk Training and Research Hospital, Izmir, Turkey Background : The aim of this study is to establish the relative frequency and distribution of neoplastic and nonneoplastic lesions of the ovarian materials sent to the Pathology Laboratory of Ataturk Research and Training Hospital, Izmir, Turkey during 2000-2005. Method : In the institutional database, during these 6 years, 938 ovarian lesions were evaluated according to histopathological diagnosis retrospectively. They were also subclassified according to the neoplastic nature, origin and localisation of the tumor . Results : Out of 938 ovarian lesions reviewed, 736 (78,46 %) were neoplastic and 202 ( 21,54 %) nonneoplastic. Of 736 neoplastic lesions, 692 (92,02 %) were primary and 44 (5,7 %) metastatic. Out of 692 primary tumors 527 (76,15 %) were epithelial, 120 ( 7,34 % ) germ cell tumor and 45 (6,50 %) sex-cord stromal tumor. Krukenberg tumor was the most frequent metastatic tumor (40 % ) noted.Out of 202 nonneoplastic lesions 80 (39 %) were endometriotic cyst. Conclusion: The data demonstrate that the profile of the studied ovarian lesions in Ataturk Research and Training Hospital quantitatively parallels most of the literature reviewed PP3-42 SUBEROYLANILIDE HYDROXAMIC ACID, A HISTONE DEACETYLASE INHIBITOR, SELECTIVELY INDUCES GROWTH SUPPRESSION AND CELL DEATH OF ENDOMETRIAL STROMAL SARCOMA CELLS IN VITRO Andelko Hrzenjak, Farid Moinfar, Marie-Luise Kremser, Bettina Strohmeier, Kurt Zatloukal, Helmut Denk Medical University Graz, Department of Pathology, Austria Background: Endometrial stromal sarcomas are rare uterine malignancies and molecular mechanisms involved in their pathogenesis are poorly understood. Beside surgical excision, very often followed by recurrences, other therapeutic methods like radio- and/or chemotherapy seem to be less efficient. Covalent modifications of histone proteins, in particular de/acetylation of lysine residues, play an important role in the regulation of gene transcription in normal and cancerous cells. Nevertheless, the studies about these processes in solid mesenchymal tumors are limited. In our recent work we have shown increased HDAC2 expression in ESS samples and cognate ESS-1 cell line in comparison to normal endometrial stroma cells. Methods: For in vitro experiments endometrial stromal sarcoma cells (ESS-1) and human endometrial stromal cells (HES) were grown under standard conditions. After treatment with suberoylanilide hydroxamic acid (SAHA) in different concentrations, the cells were harvested at 24, 48 and 72 hours and analyzed by flow cytometry and Western blotting. Cells were also continuously monitored and changes in cell-growth and morphology were documented by cell-observer. Results: Our study shows that SAHA, which inhibits the activity of histone deacetylases class I and II, is able to mediate the cell cycle and expression of some genes related to malignant phenotype of endometrial stromal tumors. We were able to show for the first time that SAHA caused a dose dependent decrease in viable cell number and an increase in percentage of dead endometrial stromal sarcoma cells over time. In our in vitro experiments SAHA (3μM) led to differentiation and to death of ESS-1-cells, but not of HES-cells. Exposure of HES-cells to SAHA resulted in slightly decreased cell proliferation without evidence of cell death. SAHA also increased the p21WAF1 expression and caused significant changes in the cell cycle by inhibiting the G1/S transition in ESS-1 cells. Recovery experiments indicated that these changes became irreversible when the tumor cells were

treated with SAHA for longer than 24 hours. Conclusion: Taken together our study suggests that HDACs might be considered as potential drug-targets in the therapy of endometrial stromal sarcomas. For different hematologic malignancies and solid tumors SAHA has recently entered the phase II and III clinical trials. Our data suggest that SAHA might be also a promising therapeutic agent for endometrial stromal sarcoma. Further studies are needed to prove its efficacy in vivo. PP3-43 EXPRESSION OF IL-32 AND ITS PROGNOSTIC RELEVANCE IN CERVICAL SQUAMOUS CELL CARCINOMA Shengjin Li1, Song Mei Huang1, Do Young Yoon2, Jin-Man Kim3 1 Department of Pathology, Chungnam National University College of Medicine, Korea 2 Bioscience and Biotechnology, Konkuk University Graduate School, Korea 3 Cancer Research Institute and Department of Pathology, Chungnam National University College of Medicine, Korea Background: IL-32 is a recently discovered cytokine that induces TNF, IL-1, IL-6, and chemokines. This study examines the expression of IL-32 and its association with clinicopathological features in cervical cancer patients. Design: The following samples were analyzed: 40 paraffin-embedded specimens of cervical squamous cell carcinoma patients treated with radiation therapy or post-operation radiation therapy. The patients were grouped into FIGO stage IB (n=16) and stage IIA-IIIB (n=24). All tissues were subjected to immunohistochemical staining for IL-32 and clinical correlation with FIGO stage and survival, and the following results were obtained. Result: IL-32 expression was observed in 55% (n=22) of all tumor tissues and was strong in the tumor invasion site. The expression was located mainly in cytoplasm as well as in some tumor cell nuclei. When IL-32 expression was investigated according to the groups with regard to the FIGO stage IB and IIA-IIIB, there was a statistically significant (Chi-square test) IL-32 expression frequency in the stage IIA-IIIB (71%) compared to stage IB (31%) (P value = 0.014). The staining intensity showed borderline significance with advanced stage (P value = 0.064). However, IL-32 expression was not correlated with survival of the patients (P value = 0.79 and 0.90 in stage IB and IIA-IIIB, respectively). Conclusion: IL-32 expression is frequently observed in cervical squamous cell carcinoma cells. Our results show that the (enhanced) expression of IL-32 by carcinoma cells is correlated with advanced stage of patients with cancer of the uterine cervix. The expression of IL-32 in itself is not associated with worse survival in these patients PP3-44 HISTOPATHOLOGICAL STUDY AND DNA IMAGE ANALYSIS IN HYDATIDIFORM MOLE AND NONMOLAR ABORTION: A STUDY OF 89 CASES IN TUNISIA Hanene Landolsi, Nabiha Missaoui, Mohamed Tahar Yacoubi, Moncef Mokni, Atef Ben Abdelkader, Sihem Hmissa, Sadok Korbi Department of Pathology, F. Hached Hospital, Sousse, Tunisia Background: Complete hydatidiform mole (CHM) and partial hydatidiform mole (PHM) are two gestational trophoblastic diseases. Commonly, CHM is genetically androgenic and diploid whereas PHM is triploid. The absence of standardized morphological criteria and the presence of atypical cases in put major problems in the histopathological recognition of these diseases. Aims: to review histological diagnosis of molar and non molar first-trimester pregnancy by two pathologists and to evaluate the correlation between the final diagnosis and some histological criteria leading to the diagnosis of mole. Then we evaluate the correlation of consensus histological diagnosis with

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ploidy status for molar pregnancy. Material and methods: We proceeded on a retrospective study of 89 specimens of abortus conception, 35 CHM, 12 PHM and 42 hydropic abortions (HA). Kappa value (K) was calculated for interobserver agreement, and to compare initial and consensus diagnosis. The final histopathogical diagnosis, was compared to the results of DNA contents detected by imaging analyser (Samba 200) studying all cases of molar pregnancy. Results: In the consensus histological diagnosis the cases were reclassified into 30 CHM (initial diagnosis (ID): 27 CHM and 3 PHM), 13 PHM (ID: 7 PHM and 6 CHM) and 46 HA (ID: 42 HA, 2 CHM and 2 PHM). There was agreement among the two pathologists in 77 cases (K= 0.72, 0.52 and 0.95 respectively for CHM, PHM and HA). We objectivate a significant statistical correlation between the final diagnosis and some histological criteria leading to the diagnosis of mole: large villi ,hydropic swelling, cistern, scalped outlines, trophoblastic hyperplasia, vacuolisation of tropholastic cells, polarisation of the trophoblastic cells, atypia and trophoblastic necrosis, free trophoblastic cells, absence of vessels and fibrosis into the villous axis. Others criteria were significantly distinctive between CHM and PHM: diffuse hyperplasia of trophoblastic cells, polarisation of trophoblastic cells and the presence of trophoblatic inclusions in the core of the villi. The ploidy results showed that 56.6 % (17/30) of CHM were diploid and 53.8% (7/13) of PHM were triploid. Conclusion: Our study demonstrates that some histopathological criteria can distinguish between PHM, CHM and non molar abortion. The study of DNA content correlate to the clinical outcome is useful in some difficult cases in which these morphological criteria were not easily reproducible. PP3-45 E-CADHERIN EXPRESSION IN ADENOMYOSIS J Julia Romadanova, Olga Popova, Julia Gorbacheva, Irina Voloshchuk Moscow Medical Academy, Russia Background. E-cadherin is one of the major molecules that determine epithelial cells adhesion and physiology. The aim of the study was to reveal the possible role of E-cadherin in ectopic loci formation in adenomyosis. Methods. E-cadherin determination using enzyme-linked immunosorbent assay was performed in endometrial and myometrial samples of 53 patients with adenomyosis and 24 women without adenomyosis. Immunohistochemical study with antibodies to E-cadherin was made on paraffin-embedded tissue sections of hysterectomy material from 12 women with adenomyosis and 4 women with benign lesions excepting adenomyosis. Results. E-cadherin concentration in endometrium of adenomyosis group was 15.95±7.72 ng/ml while in control group 8.98±4.09 ng/ml (p=0.007). In myometrium of adenomyosis group E-cadherin concentration was 1.76±1.92 ng/ml, in control group 0.66±0.27 ng/ml (p=0.05). Correlation between E-cadherin concentration and disease duration was revealed. In cases with anamnesis of adenomyosis less than 3 years E-cadherin concentration in endometrium was 8.77±2.24 ng/ml, in cases with duration of disease 4-7 years 15.96±4.36 ng/ml (p=0.002) and in cases with duration 8 years and more 24.88±13.53 ng/ml (p=0.03). In myometrium these values were 0.61±0.09, 1.64±0.99 (p=0.05), 4.61±2.99 ng/ml (p=0.01) respectively. Immunohistochemically strong membrane expression of E-cadherin was revealed in all glands both in eutopic and ectopic endometrium. Intensity of reaction was higher in ectopic loci than in eutopic endometrium. Conclusion. No evidences of homophilic adhesion disturbance of epithelial cells in adenomyosis were revealed. Increased Ecadherin expression according to disease duration allows to suppose compensatory intensification of adhesion in response to long-term existing of ectopic loci.

PP3-46 HISTOLOGICAL AND IMMUNOHISTOCHEMICAL STUDY OF SEX-CORD STROMAL TUMOURS OF THE OVARY Maria Rosa Escoda1, Irmgard Costa2, Neus Combalia2, Tamara Parra2, Jordi Antoni3, Jesus Montesinos4, Manuel Corona3, Merce Rey2 1 Department of Pathology. Corporació Parc Taulí-UDIAT, Parc Taulí s/n, 08208 Sabadell, Spain 2 Department of Pathology. Corporació Parc Taulí-UDIAT, Parc Taulí s/n, 08208 Sabadell, Spain 3 Department of Gynecology. Hospital de Sabadell-Corporació Parc Taulí, Parc Taulí s/n, 08208 Sabadell, Spain 4 Department of Oncology. Hospital de Sabadell-Corporació Parc Taulí, Parc Taulí s/n, 08208 Sabadell, Spain Background: Sex cord-stromal tumours (SCST) are rare ovarian tumours, with diverse but similar microscopic patterns, a characteristic immunohistochemical (IHC) expression and a usual benign behaviour. We analysed clinical, macroscopic, microscopic and IHC parameters, in order to better classify these tumours and to predict the prognosis. Method: 12 SCST (except thecoma-fibroma group) were retrieved from our files (19892004), and we analysed histologic features (architectural and nuclear pattern, mitosis and presence of luteinized stromal cells) and the IHC profile (CAM 5.2, inhibin, calretinin, s-actin, EMA, CK7 and neuroendocrine markers). Results: The initial diagnosis were 9 granulosa cell tumours (8 adult – AGCT- and 1 juvenile – JGCT-), 2 Sertoli-Leydig cell tumours –SLCT- (1 welldifferentiated and 1 of intermediate differentiation) and 1 unclassified SCST. The patients ranged from 16 to 75 years old (mean, 45 years), the youngest was the JGCT. The known clinical presentation was vaginal bleeding (2), pelvic pain (4) and incidental finding (1). All tumours were unilateral, solid or solidcystic (9/12) and ranged from 0,6 to 30 cm. The tumours presented an admixed variety of patterns: macrofollicular (6/12), solid (6/12), insular (3/12), microfollicular (4/12), watery-silk (2/12), and tubular (2/12). Nuclei were grooved in 10/12, and round with nucleoli in 2. Call-Exner bodies were mainly seen in microfollicular and watered-silk patterns. No significant atypia was observed and mitotic count ranged from <1/10HPF to 12/10HPF. The tumours stained positively for calretinin (8/8), sactin (5/8), CAM5.2 dot-like (6/8) and diffuse (2/8) and inhibin (1/8). All were negative for EMA, CK7 and neuroendocrine markers. Luteinized stromal cells were highlighted with inhibin and calretinin. We reclassified the well differentiated SLCT as Sertoli cell tumour (SCT) and the SLCT of intermediate differentiation and the unclassified SCST as AGCT. Only the SCT and one AGCT presented pelvic-abdominal recurrences 9 and 15 years later, respectively. The mitotic activity (6/10 HPF) and the poorly defined margins seemed to be the prognostic factors in the SCT. The recurrent AGCT corresponded to the largest tumour in our series. Conclusions: SCST exhibit several growth patterns. The insular, solid and tubular patterns raise the most differential diagnosis among the SCST and with other ovarian neoplasms. Nuclear grooves (hallmark of the GCT) and the IHC profile described will help in establishing the diagnosis. Some histologic features could be associated with an adverse outcome. PP3-47 ENDOMETRIAL CARCINOMA WITH “PREVIOUS INADEQUATE SPECIMEN SAMPLING” IN ENDOMETRIAL BIOPSY Ana Blasco1, Laura Yébenes1, Gabriel Olmedilla1, Paloma Ramos1, Pedro Valenzuela2, Antonio Ruíz1 1 Department of Pathology, Príncipe de Asturias University Hospital, Alcalá de Henares, Madrid, Spain 2 Department of Obstetrics and Gynecology, Príncipe de Asturias University Hospital, Alcalá de Henares, Madrid, Spain

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BACKGROUND: Endometrial biopsy reported by pathologists as “inadequate specimen sampling” (“insufficient material” or “no representative sampling”) can lead to an erroneous idea of absence of malignancy. The aim of this study is to find out the overall rate of biopsies diagnosed as “inadequate sampling”, and its repercussion on the time-delay in establishing the diagnosis of endometrial cancer. METHOD: Clinical data and pathological reports from 227 patients with endometrial carcinoma, over a period of fourteen years (from 1989 to 2003) were retrospectively reviewed, and the “insufficient” or “no representative” samples proportion in endometrial biopsies, prior to histological confirmation of malignancy, was calculated. RESULTS: Eighteen out of these 227 patients (7.9%) had a previous pathological report of “inadequate sample for diagnosis”. Only two of those eighteen cases (11.1%) underwent a new biopsy in less than a month, and twelve (66.6%) had a delay in the rebiopsy of more than three months. CONCLUSIONS: Assessment of specimen adequacy is important for pathologist interpreting endometrial biopsies. Endometrial biopsies reported as “inadequate sample for diagnosis” should not be considered, for practical purposes, as a benign sample, and early rebiopsy should be recommended. PP3-48 EXPRESSION OF SURVIVIN AND TOPOISOMERASE II ALPHA IN OVARIAN CARCINOMA: CORRELATION WITH CLINICOPATHOLOGICAL PARAMETERS AND PROGNOSIS Derya Gumurdulu1, E. Handan Zeren1, Baris Guzel2, Gulsah Seydaoglu3, Seyda Erdogan1, Suzan Zorludemir1, Mehmet Ali Vardar2, Aytekin Altintas2 1 Cukurova University, Faculty of Medicine, Department of Pathology, Adana, Turkey 2 Cukurova University, Faculty of Medicine, Department of Obstetrics and Gynecology, Adana, Turkey 3 Cukurova University, Faculty of Medicine, Department of Biostatistic, Adana, Turkey Background: Ovarian carcinomas account for four percent of all female cancers and for 25% of cancers of female genital organs. The clinicopathological parameters, such as clinical stage, tumor grade, histological type may reflect the aggressiveness of ovarian tumors and clinical outcome. The additional prognostic information closely related to tumor cell biology is essential for the identification of patients with poor prognosis. Survivin is a member of the inhibitor of apoptosis protein (IAP) family that has been reported to be broadly expressed in a wide range of malignant tumors but not in most corresponding normal tissues. Topoisomerase IIĮ (T2a) is an enzyme with an important role in DNA topology, repair and replication. It is a cell-cycle-related protein, expressed in normal as well as neoplastic cells in the S, G2 and M phases. The aim of this study was to investigate the association of survivin and T2a expression with clinicopathological parameters and prognosis. Materials and methods: Formalin-fixed, paraffin-embedded tissue sections from 61 ovarian carcinomas were obtained from the files of the Cukurova University, Faculty of Medicine, Department of pathology. We performed primer antibodies to survivin, T2a and Ki67 by immunohistochemically. Nuclear staining for all antibodies were considered as positive reactivity, percentage of positive cells was scored on a three-tiered system with <10% as negative. and we evaluated the relationship between expressions of survivin, T2a, Ki67 and prognosis. Results: The age of patients ranged from 17 to 78 and the mean age was 52.16 ± 14.84. The 61 cases in the study comprised 35 serous carcinomas,, 12 endometrioid carcinomas, 6 mucinous carcinomas, 2 clear cell carcinoma and 6 mixed carcinoma of the ovary. Survivin expression was found in %27.9 (17 of 61) of the tumors. T2a positivity was noted in %91.8 (51 of 61) of the tumors. There was a correlation between Ki 67 and T2a expression (r=0.28, p=0.030). Survivin and T2a expression did not correlate

histological type, stage or overall or disease-free survival. Conclusion: The immunohistochemical assessment of survivin and T2a does not seem to be helpful in the prognostic characterisation of ovarian carcinoma. The most important prognostic factors are stage and histological type. Further studies addressing to find the additional prognostic parameters will likely provide valuable insights into the behaviour of ovarian carcinoma. PP3-49 IMMUNOHISTOCHEMICAL STUDY OF THE EXPRESSION CD44 IN SEROUS, BORDERLINE, MALIGN SURFACE EPITHELIAL OVARIAN TUMORS AND METASTATIC OVARIAN TUMORS Gamze Yurdakan1, Nilufer Onak Kandemir1, Sacide Colak1, Aylin Ege Gul2, Nimet Karadayi2, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Dr. Lutfi Kirdar Kartal Research and Training Hospital, Department of Pathology Istanbul, Turkey Backround: Carcinogenesis and metastasis are a multistep process involving complex interactions between tumor cells and the environment. The main cause for the motion of the tumor cells in invasive carcinomas may be loss of completeness intercellular adherence junction. One of these adhesion receptors, CD44 binds to extracellular matrix component hyaluronic acid. In the present study, to elucidate role of CD44, we investigated a series of benign, borderline, and malignant ovarian serous neoplasms. Materials and Methods: Archives paraffin- embedded formalin-fixed blocks from benign serous tumors (n=11), benign mucinous tumors (n= 8), borderline serous tumors (n=6), borderline mucinous tumors (n=1), primer malign ovarian serous tumors (n=9) and metastatic ovarian tumors (n=12) were immunohistochemically stained for CD44. Percentage of reactive tumor cells and stromal cells with CD44 were scored. Positive staining in at least 10% of cells with strong staining considered being positive. Staining intensity was scored from 1+ to 3+. CD44 protein was preferentially expressed along the basolateral domain of plasma membrane of tumors cells. Some of tumor cells were reactive with CD44 in cytoplasm. Results: CD44 was not only detected in 1 (5, 2%) of benign tumors. Remain of the tumors were reactive with CD44 in differential percentage and localization. CD44 staining was evaluated in 4 (8, 33%), in 21 (43, 75%), in 16 (33, 33 %), in 7 (14, 58%) of all tumors as grade 0, 1, 2, 3 respectively. Statistically significant associations were found between serous and mucinous benign tumors (p=0.0001), borderline and malign serous tumors (p=0.001), malign serous tumors and metastatic carcinomas (p=0.054), and primary malign ovarian tumors and metastatic carcinomas (p=0.002) as compared CD44 staining grade. Grade 0,1,2,3 stromal staining was determined in 14 (29, 16%), in 31 (64, 58 %), in 1 (2, 08 %) and in 1 (2, 08 %) ovarian tumors respectively. But no difference was in reaction with CD44 in stromal cells statistically. In primary malign tumors, CD44 was detected more than primary benign ovarian tumors and was different statistically (p=0.0001). Conclusion: These results suggest that CD44 expression may be important in differential diagnosis between borderline and malign serous tumors, primary malign ovarian tumors and metastatic carcinomas. Also CD44 expression is a characteristic factor in stromal invasion for ovarian serous carcinomas. A large number of samples are necessary to verify prognostic significance of CD44 expression in tumor progression.

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PP3-50 COMPARISON OF TWO VERSUS THREE GRADES FOR ENDOMETRIAL CARCINOMA: AN ASSESMENT OF INTEROBSERVER RELIABILITY IN ENDOMETRIAL SAMPLES AND HYSTERECTOMY SPECIMENS Sema Ozuysal, Ilker Ercan, Hulya Ozturk Nazlioglu, Ulviye Yalcinkaya, Berna Calisir Uludag University School of Medicine Bursa, Turkey OBJECTIVE:To compare the agreement in determining the histological grade of endometrial adenocarcinomas in endometrial samples and hysterectomy specimens using the three-grade FIGO classification and the two-grade system. METHODS: Endometrial samples and subsequent hysterectomy specimens from 55 patients with endometrioid adenocarcinoma were graded according to the FIGO and the binary system (BS) proposed by Lax et al. All tumours were graded independently by four pathologists on two separate occasions. The reliability coefficient for two-facet crossed design (specimen x observer x occasion) based on generalizability theory was calculated for each grading system in endometrial samples and hysterectomy specimens. We used the generalizability coefficient (GC) as a measure of interobserver reliability and the factors which could influence it (different psychological status of the observers at different times, changes in the laboratory conditions, etc).GC was interpreted as follows: 0.00 to 0.40, poor; 0.41 to 0.60 low; 0.61 to 0.80 moderate; and 0.81 to 1.00, high. The pathologists were separated into two groups as “experienced in gynecologic pathology” and “with limited experience in gynecologic pathology”. Exact agreement percentages were calculated as the percent of all cases in which the grades were identical in endometrial samples and hysterectomy specimens. The FIGO and BS were compared by using chi–square test and kappa values were calculated for each grading system. RESULTS: GCs were 0.93 for endometrial samples, and 0.83 for hysterectomy specimens for FIGO system. The BS revealed GCs of 0.89 and 0.83 for endometrial samples and hysterectomy specimens, respectively. The GC levels were higher for experienced pathologists both when using the FIGO (endometrial samples= 0.92 hysterectomy samples= 0.92), and the BS (endometrial samples=0.92 hysterectomy samples= 0.90) compared to the group with limited experience in gynecologic pathology (FIGO0.89- 0.49; BS- 0.68- 0.60). The agreement rates between the endometrial sampling grade and the hysterectomy grade in FIGO and BS were 247/439 (%56.2) and 361/439 (%82.2) respectively (p<0.05). Kappa values were 0.29 (p<0.05) in FIGO system and 0.38 (p<0.05) in BS. CONCLUSION: In this study high GC values were obtained both for the FIGO and BS. GC levels were higher for experienced pathologists in both the FIGO and BS than the group with limited experience in gynecologic pathology. The agreement rates between the endometrial sampling grade and the hysterectomy grade were higher in the BS compared to the FIGO system. PP3-51 BABES-PAPANICOLAOU TEST METHOD TO VERIFY LSIL TREATED LESIONS Eliza Gramada, Geanina Micu, Ionela Celea, Adina Ene, Andrei Murariu, Florica Staniceanu Colentina University Hospital, Department of Pathology, Bucharest, Romania Background: The quality verification of the treatment applied by the gynecologist is realised by periodical cytological cervicovaginal examinations using Babeú-Papanicolaou test. This test may indicate the presence or the absence of pathologycal cells. Material and method: In the Department of Pathology from Colentina Universitary Hospital, the authors took for study a lot of 83 patients, with the age between 20-30 years, treated by the gynecologist with Podofilin or electrocautherisation for mild dysplasia and plan condyloma. The study analyzed the koilocytic

cells, the intermediate squamous epithelial cells with nuclear increased size and/or mild dyscariosis, pathological changes present in mild dysplasia and plan condyloma. The smears were stained with Papanicolaou stain and the 2001-revised Bethesda System was used in order to interpret the results. Study results: From all the treated patients, 69 patients were present for a control test (63,13%), 52 of them (75,36%) being treated for plan condyloma. Among these, using the cervico-vaginal examinations, 15 cases (28,84%) have had normal aspect, 9 cases(17,30%) presented inflammatory aspects of the reparing type, 28 cases (53,84%) presented koilocytic cells. From the 17 cases (24,67%) with mild dysplasia, a number of 11 cases (64,70%) presented normal Pap smears, and 6 cases (35,30%), presented ASC-US subtype modifications. Conclusion: The study displays that, from the number of the patients examinated, almost 40% completely recovered while the other 60% remained with pathological modifications. These data prove the necessity of cytodiagnostic reevaluation after treatment. PP3-331 UTILIZING OF KI 67 EXPRESSION AND ESTROGEN AND PROGESTERONE RECEPTORS IN DIFFERENTIAL DIAGNOSIS OF SMOOTH MUSCLE TUMORS OF THE UTERUS Nilgun Dicle, Sevil Sayhan, Duygu Ayaz, Sibel Kececi, Hakan Camizoglu Ege Dogumevi ve Kadin Hastaliklari Egitim Hastanesi, Izmir, Turkey BACKGROUND: Smooth muscle tumors of the uterus are frequent and most of them are benign. Some leiomyomas may have unusual morphologic features difficult to distinguish from leiomyosarcomas. Smooth muscle tumors of uncertain malignant potential (STUMP) represent a variant of uterine leiomyomas and remain a dilemma due to their uncertain clinical behavior. MATERIAL AND METHODS: One thousand four hundreds and twenty patients operated for smooth muscle tumor of uterine origin between 2005-2007 at our hospital’s department of gynecology. Seven leimyosarcoms and 7 STUMP cases were reported during this term. The average age of STUMP and leiomyosarcoma cases was 42 and 57 years respectively. The average size of STUMP and leiomyosarcoma cases was 6.8 and 12 cm respectively. Seven leiomyomas with similar age and sizes were selected to compare with STUMP cases. Number of mitosis, presence of necrosis, atypia, Ki 67 expression, presence of estrogen and progesterone receptors were evaluated in paraffin embedded tissue. CONCLUSION: There is limited experience in the clinical behaviors of problematic uterine smooth muscle tumors like STUMP. Several parameters are used to facilitate the appropriate diagnosis. Besides the criteria of the WHO classification we found that the expression of Ki 67 and the presence of estrogen and progesterone receptors in uterine leiomyomas, STUMP and leiomyosarcomas is helpful in differential diagnosis.

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Uropathology PP3-52 E-CADHERIN EXPRESSION IN TUR MATERIALS WITH LOW AND HIGH GRADE UROTHELIAL CARCINOMAS: A STUDY OF 30 CASES Elif Ozer, Derya Arslanoglu, Haluk Pulat, Mehmet Gonultas, Muzaffer Caydere, Hesna Muzeyyen Astarci, Huseyin Ustun Ankara Education and Research Hospital, Pathology Department, Turkey The cadherins are a family of transmemrane glycoproteins and are the primer mediators of intercellular adhesion. Cadherins are responsible for cell adhesion and transmembrane protein and expressed in all tissues. E-cadherin, the epithelial spesific cadherin mediates the selective adhesion of epithelial cells and required for the interaction and maintenance of normal epithelial integrity. Loss of E-cadherin expression leads to dissociation of cells from cohesive tissues and generates dedifferentiation and invasiveness in a variety of solid tumors, revealing the crucial role of E-cadherin as a suppressor of tumor invasion and metastasis. This study comprised with 30 transurethral resection of urinary of bladder tumor specimens. We immunostained 15 cases of low grade papillary urothelial carcinomas and 15 cases of high grade papillary urothelial carcinomas for E-cadherin using tissue arrays. We observed that E-cadherin expression in low grade tumors is more pronounced than high grade malignancias. In 5% of low grade tumors E-cadherin expression was indistinct than aspected. We think that this must be due to the increased potential of invasiveness and metastasis, contrary to their histological features. We propose to use E-cadherin as a prognostic marker in TUR materials for urothelial carcinomas. PP3-53 RELATIONSHIP BETWEEN GLEASON SCORE AND NUCLEAR SIZE AND SHAPE FACTORS IN PROSTATIC ADENOCARCINOMA Sibel Bektas1, Burak Bahadir2, Banu Dogan Gun2, Gurkan Kertis2, Sukru Oguz Ozdamar2 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey Background: The Gleason grading system which depends on differentiation of the glandular structures is the most common used grading system in prostatic adenocarcinoma. Recently several quantitative morphometric methods have been developed to test the objectivity of the Gleason grading system. The aim of this study is to assess the size and shape characteristics of the tumor cell nuclei, and compare the results of measurement with Gleason score of the cases by using computer-assisted analysis system. Methods: This study included 130 cases with prostatic adenocarcinoma, 100 (77%) were needle biopsy specimens and 30 (23%) were prostatectomy specimens. The ages of patients ranged 46 to 94 (Mean: 67±8.26). Each slide were reevaluated and rescored by the same pathologist without having any knowledge about the prior diagnoses. About 150 nuclei with sharply demarcated contour were included for morphometric analysis for each case. After transferring microscopic images to the computer, nuclear area, nuclear length, nuclear perimeter, roundness factor, and form ellipse were automatically measured by an image analysis program. The relationship between Gleason score and these variables were determined by correlation analysis with Spearman and Pearson correlation coefficient. Results: Among all the specimens, Gleason score were significantly correlated with mean nuclear area (r: 0.516, p=0.01), with mean nuclear length (r: 0.298, p=0.01) and with mean nuclear perimeter (r: 0.303, p=0.01). However, in the needle biopsy group Gleason score were significantly correlated with mean nuclear area (r: 0.522, p=0.01), with mean nuclear length (r:

0.398, p=0.01), with mean nuclear perimeter (r: 0.432, p=0.01) and with mean form ellipse negatively (r: -0.213, p=0.05). In the prostatectomy group Gleason score were correlated with mean nuclear area (ı: 0.619, p=0.01) and mean roundness factor negatively (ı: -0.425, p=0.05). Conclusions: As a result, nuclear size and shape factors especially mean nuclear area were found concordant with Gleason score. Nuclear size and shape assessment may support to evaluate pathologic status of the disease in the prostatic adenocarcinoma. On the other hand, the association of clinical outcome and morphometric parameters could not be evaluated because of the short observation period in this study. Therefore, more detailed studies with long period of observation are required to asses more accurate results. PP3-54 INTRAOBSERVER AND INTEROBSERVER REPRODUCIBILITY OF THE FUHRMAN NUCLEAR GRADING FOR RENAL CELL CARCINOMA Sibel Bektas1, Figen Barut1, Burak Bahadir2, Banu Dogan Gun1, Nilufer Onak Kandemir1, Aylin Ege Gul3, Sevinc Keser3, Nimet Karadayi3, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 3 Lutfi Kirdar Kartal Training and Research Hospital, Department of Pathology, Istanbul, Turkey Background: Several systems have been proposed for grading renal cell carcinoma. Classification of renal cell carcinoma has been divided into four categories according to nuclear size and the presence of nucleoli, the most widely used grading system reported by Fuhrman. However, there are few studies in the literature assessing the intraobserver and interobserver variability of this system. The objective of the current study was to evaluate the intraobserver and interobserver reproducibility of the Fuhrman nuclear grading system. Methods: In this study, pathology slides from 46 cases of renal cell carcinoma (Conventional: 37, chromophobe: 5 and papillary: 4) were rescored according to Fuhrman nuclear grading system by two pathologists. The intraobserver variation was determined as the first pathologist’s observation on two different occasions. Interobserver variation was determined by the second pathologist who did not known the result of the first. Both intraobserver and interobserver reproducibility were assessed using Cohen’s kappa statistics. Results: The original Fuhrman nuclear grade was grade 1 in 4 patients (8.7%), grade 2 in 30 patients (65.2%), grade 3 in 11 patients (23.9%), and grade 4 in 1 patients (2.2%). After slides review by same pathologist, nuclear grades were reassigned as follows: grade 1 in 8 patients (17.4%), grade 2 in 23 patients (50%), grade 3 in 14 patients (30.4%), and grade 4 in 1 patients (2.2%). Intraobserver reproducibility of the Fuhrman nuclear grading system was found to be substantial (ț = 0.66). Fuhrman nuclear grading of the second pathologist was grade 1 in 11 patients (23.9%), grade 2 in 27 patients (58.7%), grade 3 in 7 patients (15.2%), and grade 4 in 1 patients (2.2%). Interobserver reproducibility of the Fuhrman nuclear grading system was found to be moderate (ț = 0.42). We found low intraobserver and interobserver agreement for the Fuhrman nuclear grade 2 and grade 3, respectively. Conclusions: Despite strong intraobserver reproducibility of the Fuhrman nuclear grading system, one should consider moderate interobserver reproducibility and low agreement for the grade 2 and grade 3. Further studies are required to confirm our results in larger series.

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PP3-55 CONCORDANCE OF NUCLEAR MORPHOMETRIC ANALYSIS WITH FUHRMAN NUCLEAR GRADING IN CONVENTIONAL RENAL CELL CARCINOMA Sibel Bektas1, Figen Barut1, Gurkan Kertis1, Burak Bahadir1, Banu Dogan Gun1, Nilufer Onak Kandemir1, Nagehan Ozdemir2, Nimet Karadayi, Sukru Oguz Ozdamar1 1 Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey 2 Lutfi Kirdar Kartal Training and Research Hospital, Department of Pathology, Istanbul, Turkey Background: Nuclear grade, stage and tumor size are generally used for estimating prognosis of renal cell carcinoma. Ouantitative image analysis has been applied to renal cancers for diagnostic and prognostic purposes. The aim of this study was to investigate whether there is any correlation between the results of morphometric measurement and Fuhrman nuclear grading score, pathologic stage and size of conventional (clear cell) renal cell carcinoma. Methods: Morphometric nuclear parameters such as mean area, mean roundness factor, mean form ellipse, mean length, mean breadth and mean perimeter were evaluated in hematoxylin and eosin stained slides of 37 conventional renal cell carcinoma by using computer assisted image analysis system. The relationship between Fuhrman nuclear grading score, pathologic stage, tumor size and morphometric results were determined by correlation analysis with Pearson correlation coefficient. Results: Mean nuclear area were correlated with pathologic stage (r: 0.413, p=0.05) and with Fuhrman nuclear grade (r:0.588, p=0.01). Mean nuclear length were correlated with pathologic stage (r:0.446, p=0.01) and with Fuhrman nuclear grade (r:0.580, p=0.01). Mean nuclear breadth were correlated with pathologic stage (r:0.377, p=0.05) and with Fuhrman nuclear grade (r: 0.544, p=0.01) and with tumor size (r:0.366, p=0.05). Mean nuclear perimeter were correlated with pathologic stage (r: 0.449, p=0.01) and with Fuhrman nuclear grade (r:0.593, p=0.01). Mean nuclear roundness factor were correlated with pathologic stage (r: 0.418, p=0.05) and with Fuhrman nuclear grade (r: 0.456, p=0.01). The correlation between mean nuclear form ellipse and Fuhrman nuclear grade, pathologic stage, and tumor size was not statistically significant. Conclusions: In this study, the relative concordance of morphometric results and pathologic stage, Fuhrman nuclear grade, and tumor size exhibited the importance of nuclear morphometric analysis in conventional renal cell carcinoma. Studies including larger series of cases investigating detailed nuclear morphometric analysis of conventional renal cell carcinoma are needed. PP3-56 CHROMOPHOBE RENAL CELL CARCINOMA Nadia Kourda, J Kourda, R Noomene, A Derrouiche, O Lamine, M Ferchiou, M Chebil, Sara Baltagi Ben Jilani, R Zermani Charle Nicolle’s Hospital Department of Pathology, Tunisia INTRODUCTION: Chromophobe renal cell carcinoma (CRCC) is an uncommon,which accounts for average 5% of all renal neoplasm. CRCC shows an intermediate prognosis between conventional cell carcinoma and oncocytoma. Oncocytoma, a benign renal tumour, is considered the aim histological differential diagnosis of CRCC. A careful anotomopathologic study, special coloration and immunostaining, allow differentiating these two entities. We report 21 cases of CRCC. The aim of our study was to evaluate the incidence, clinical presentation, anatomopathological and immunohistochemical findings, prognosis, and clinical outcome of CRCC. METHODS: Hospital records and pathological slides were reviewed for 21 patients CRCC treated at Charles Nicolle Hospital between 19992006. The relevant clinical and pathologic data were extracted from the clinical charts and collected into a unique database. The following parameters were noted in each patient: age, sex,

symptoms at diagnosis, tumour diameter, TNM stage and grade, histological cell type, follow-up time, local recurrence, disease progression, and death. RESULTS: 21 cases of CRCC; 8 (38%) were men and 13 (62%) women. The mean patient age was 54.8 (range 31to 74 years). The tumour was located in the right kidney in 11 patients (53%) and in the left kidney in 10 (47%); no familial or bilateral disease was observed. An incidental diagnosis was founded only in two cases and accounted for 9.5% of the cases. The other patients were symptomatic. The mean tumour size was 7,9 cm (between 2,5 to 16,5cm). Grossly, it was in 95%, a mediocortical and a necrotico-hemorragic tumour. On microscope, most of the pathologic features were typical. However, in 24% we needed immunostainnig using cytokeratine 7, vimentine, CD117 and PS100. At diagnosis, 10 patients (48%) had Stage pT1a, 1 (4.7%) pT1b, 5 (24%) pT2, 1 (4.7%) pT3a, 3 (14%) pT3b, and 1 (4.7%) pT4. The lymph node status was pN0 in 19 (90.5%), and N1 in only 2 patients (9.5%). Of the 21 patients, there were no metastatic disease at diagnosis, and 1 had pulmonary metastasis. The nuclear grade was, grade 1 in 1case, grade 2 in 17 (81%), and grade 3 in 3 (14%). Radical nephrectomy was performed in 20 patients (95%). After a median follow-up of 3 years (Range 2 to 5), one local recurrence was observed. The 5-year overall survival rate for CRCC was 90%. Of the 21 patients, 3 died of unrelated causes and renal cancer, respectively. CONCLUSION: The results of our study have confirmed a general favourable outcome for CRCCs, which are predominantly low-stage and low-grade tumours. These tumours had little local aggressiveness. Although metastases at diagnosis and disease progression after nephrectomy are rare, this neoplasm should be distinguished from renal oncocytoma. PS100 A1 seems to be a powerful marker to differentiate these two entities.CONCLUSION: The results of our study have confirmed a general favourable outcome for CRCCs, which are predominantly low-stage and low-grade tumours. These tumours had little local aggressiveness. Although metastases at diagnosis and disease progression after nephrectomy are rare, this neoplasm should be distinguished from renal oncocytoma. PS100 A1 seems to be a powerful marker to differentiate these two entities. PP3-57 RENAL ONCOCYTOMA. PATHOGICAL FINDINGS Nadia Kourda, O. El Lamine, R. Noomene, A. Derrouiche, J. Kourda, M. Ferchiou, M. Chebil, Sarrah Baltagi Ben Jilani, R. Zermani Charle Nicolle’s Hospital Department of Pathology, Tunisia INTRODUCTION: We report the clinical and histopathological features of 10 cases of oncocytoma. One diagnostic problem is distinguishing oncocytoma from other renal epithelial tumours with pink cytoplasm. Oncocytomas are the most common benign epithelial tumours. It accounts about 10% of renal tumours. MATERIALS AND METHODS: Hospital records and pathological slides were reviewed for 10 patients with renal oncocytoma treated at Charles Nicolle Hospital between 19992006. The clinical data included age and gender. Microscopic examination was performed on all cases. RESULTS: The median age of the patients was 54,9 years (range 23-84 years). All our patients were symptomatic (10/10) flank pain (8/10) hematuria (2/10). They occur more frequently in women than in men (Female’s predomination (9/10)) and have a peak incidence in the seventh decade as same as RCC. On gross examination, most of the tumours were cortical in 6 cases, and the median tumour size was 74 mm (range 30-120 mm). Oncocytomas were solitary renal tumours in the ten cases we study. Multiple and/or bilateral cases were not observed. Most of these tumours were wellcircumscribed encapsulated (6/10) neoplasm, with a uniform mahogany (6/10) or red dark color (4/10). A central, white satellite scar was seen in 3 cases. Necrosis (3/10), hemorrhage (2/10). Histollogically, different architectural pattern was found with nesting (7/10), acini (4/10) and/or“tubulocystic” (4/10) architecture with a myxoid and/or hyalinized stroma (3/10). A

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moderate to abundant amount of eosinophilic cytoplasm with central, small, oval–round nuclei is found in all our cases. DISCUSSION: Light microscopic differentiation of oncocytomas from RCC specifically eosinophilic variants of the chromophobe type can be very challenging. The nuclear features are an important discriminating feature; RCCs have wrinkled, raisinoid nuclear membranes, whereas oncocytomas have round nuclei with nucleoli. Additional staining techniques and immunostaining may be helpful. In fact, Hale's colloidal iron positive staining is strong and diffuse in chromophobe RCC, whereas oncocytomas exhibit focal, weak positivity. Cytokeratin 7 is often negative in oncocytoma. Conclusion: Based on our findings, oncocytomas and RCC have similar clinicopathologic features. On the other hand, the nuclear features are an important discriminating feature; RCCs have wrinkled raisinoid nuclear membranes, whereas oncocytomas have round nuclei with nucleoli, immunohistochemical stain has been helpful in some studies. Knowledge of these distinguishing histological and immunohistological features will enable to better differentiate these two pathologic entities. PP3-58 THE EFFECT OF ORAL HIGH ALUMINIUM INTAKE ON RAT SPERMATOGENESIS Iran Rashidi1, Ali Reza Mozaffari2 1 Pathologist. Head of Department & Member of Expert Committee, Iran 2 M. D. – Internist, Iran ABSTRACT: Introduction : Aluminum is a silver-white flexible metal with a vast number of uses. It is poorly absorbed and efficiently eliminated; however, when absorption does occur aluminum is distributed mainly in bone, liver, testes, kidneys, and brain ( 1 ) . Aluminium poising causes wide range of disorders, including: a decrease in the release of neurotransmiters and inhibition of voltage dependent calcium channels. The role of calcium on GnRH release and its action is detected so, in this studying, the effect of high aluminium intake on rats spermatogenesis is investigated. Methods and Materials: The experiment performed in four groups, a control group and three experimental groups consumed 0.625, 1.25 and 2.5 mg aluminium per gram diet for 60 days. Epididymis and vas deferens were dissected cut and diluted with normal salin. In all groups weight of vas deferens , epididymis, testis and whole animal, sperm count per gram deferens and epididymis tissues were determined then, the testicular tissues fixed in formalin for study of histopathology. Results: The results have shown that in experimental groups which consoumed 1.25 and 2.5 mg aluminium per gram diet, the vas deferens, epididymis, testis and animal weight were significantly decreased. In this animals the number of sperm per gram tissues from vas deferens, epididymis were reduced. The maturation arrest is seen in seminoferous duct and it haven‘t spermatogenesis. Conclusion: This studying indicated that high aluminium in take in rat have an inhibiting effect on spermatogenesis and this effect is dose dependent. PP3-59 COMPARISON OF HIF-1Į IMMUNREACTIVITY WITH TUMOR STAGE, GRADE, ANGIOGENIC PROFILE AND PROLIFERATIVE INDEX IN BLADDER UROTHELIAL CARCINOMAS Hale Deniz1, Metin Karakok2, Faruk Yagci3, Muhammet Emin Guldur4 1 Gaziantep Pediatric Hospital, Turkey 2 Gaziantep University, Medical Faculty, Department of Pathology, Turkey 3 Gaziantep University, Medical Faculty, Department of Urology, Turkey 4 Gaziantep Gynecologic and Obstetric Hospital, Gaziantep, Turkey

BACKGROUND: Hypoxia-inducible factor-1Į (HIF-1Į) is a critical regulatory protein of cellular response to hypoxia. In this study, we evaluated the relationship of HIF-1Į immunreactivity with clinicopathologic parameters such as tumor stage and grade, as well as angiogenic profile and proliferation index. METHOD: The expression of HIF-1Į was assesed in 70 primary bladder urothelial carcinoma tissue samples immunohistochemically. Vascular endothelial growth factor (VEGF) immunreactivity and microvessel density (MVD) were used to evaluate the angiogenic profile. MVD was calculated by immunohistochemical staining of endothelial cells with CD34. Proliferation index was determined by the percentage of Ki-67 nucleer staining in tumor cells. RESULTS: There was a significant relationship between HIF-1Į immunreactivity and stage, as well as histological grade of the tumor (p<0,001). HIF-1Į immunreactivity was also closely related to VEGF expression (p<0,001), MVD (p=0,002) and proliferation index (p<0,001). VEGF, MVD and proliferation index were found to be closely related to tumor stage and histological grade. There was no correlation between HIF-1Į immunreactivity and lamina propria (p=0,13), muscularis propria (p=0,009) or vascular invasion (p=0,1). CONCLUSION: In this study, HIF-1Į expression was found to be closely related to prognostic parameters (stage, grade, angiogenic profile and proliferation index) in urothelial carcinoma of the bladder. For this reason, it may be a useful marker to determine the prognosis and to choose the appropriate treatment modality. PP3-60 IMMUNOHISTOCHEMICAL EXPRESSION OF P53, P16 AND P21 IN UROTHELIAL CELL CARCINOMA AND THEIR CORRELATION WITH CLINICOPATHOLOGICAL FEATURES Adamandia Zizi-Sermpetzoglou, Xanthippi Grammatoglou, Chryssoula Glava, Maria-Eleni Nikolaidou, Nerantzoula Petrakopoulou, Thivi Vasilakaki Department of Pathology, Tzaneion General Hospital of Piraeus, Piraeus, Greece Background: Transitional cell carcinoma (TCC) comprises approximately 90% of all primary tumors of the urinary bladder. Several types of mutations of cell cycle regulatory genes are commonly in those tumors. Alterations in the p53 are a predominant component in the TCC. Recently another gene – INK4A, which is located in chromosome 9p21, encodes proteins p16 and p19 which regulate the pRb and p53 pathways. P21 is both a p53-inducible and p53-indipendent cyclin-dependent kinase inhibitor that can arrest the cell by inhibiting DNA replication. The aim of this study was to determine the association of p53, p21 and p16 expression with clinocopathological features. Materials and methods: Immunohistochemical staining for p53, p16 and p21 was carried out on serial sections from (archival) specimens of 108 patients who underwent transurethral resection for TCC. Based on percentage of nuclear reactivity, p53 was considered as wild-type (0% to 10%) or altered (>10%); p21 was scored as wild-type (>10%) or altered (<10%); and p16 status was considered wildtype (1% to 50%) or altered (0% or >50%). Results: Positivity of p53 was detected in 80/108 (81,5%). P21: absence or focal positivity (<10%) of tumor cells was detected in 72/108 (66,7%) of the carcinomas. Absence of p16 was detected in 40/108 (37%) of the tumors. The p53 phenotype seems to be the strongest predictor of bladder cancer associated with bladder cancer progression. P21 expression was correlated with tumor grade and stage. P16 expression was related to recurrence and survival but not to tumor progression. The alteration of each marker was independently associated with disease progression (p[LTEQ]0,30) and disease-specific survival ( p[LTEQ]0,37 ). Conclusion: This study suggests that p53 and p21, as well as p53 and p16, have cooperative and synergistic effects in the promotion of bladder cancer progression.

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PP3-61 PROGNOSTIC VALUE OF EGFR AND HER 2 IN UROTHELIAL CARCINOMA Adamandia Zizi-Sermpetzoglou, Xanthippi Grammatoglou, Maria-Eleni Nikolaidou, Nerantzoula Petrakopoulou, Chryssoula Glava, Thivi Vasilakaki Department of Pathology, Tzaneion General Hospital of Piraeus, Piraeus, Greece Introduction: Bladder cancer is a common malignancy with highly growth patterns. HER 2 is one of the most frequently amplified oncogenes and a potential therapeutic target in bladder cancer. EGFR has been associated with genes of bladder tumor. The aim of this study was to investigate the expression of EGFR and HER 2 and examine their prognostic value in the progression of bladder cancer. Materials and methods: We examined the immunohistochemical expression of HER 2 and EGFR in formalin-fixed paraffin samples from 109 cases of urothelial carcinoma (TUR) corresponding in 77 cases of invasive (stage pT1-4, grade 1 to 3) and 32 cases of noninvasive (pTa, grade 1 to 3) carcinoma. Results: EGFR expression was stronger in invasive tumors (p<0,0001) and in high grade tumors (<0,0001). 52 of the 77 investigated invasive (67,5%) and 26 of the 32 noninvasive (81,3%) carcinomas showed overexpression of HER2 protein. Of 77 invasive carcinomas 23 (29.9%) showed 3+ positivity , 29 (37.7%) showed 2+ positivity and 25 (32,4%) were scored as negative with a score of +1 and 0. Of the 32 cases no sample was evaluated as 3+ positive, 26 (81,25%) showed a 2+ positivity and 6 (18,75%) cases were identified as 1+ positive or negative. Conclusion: The combination of HER 2 and EGFR or the correlation between these markers and histological grade will yield prognostic information that is more powerful than what histologic grade alone can provide. The staining pattern of EGFR and its widespread expression in bladder cancer make it a good target for antigrowth factor or gene therapy. The validation of over expression of HER 2 oncogene in bladder cancer may allow for the potential use of Herceptin antibody therapy. PP3-62 CAVERNOUS HEMANGIOMA OF THE BLADDER. A CASE REPORT Nerantzoula Petrakopoulou1, Xanthippi Grammatoglou1, Chryssoula Glava1, Spiridon Koutsoukis1, Maria Politou2, Thivi Vasilakaki1 1 Department of Pathology, Tzaneion General Hospital of Piraeus, Piraeus, Greece 2 Department of Pathology, General Hospital of Rhodos, Greece Objectives: Bladder hemangiomas are exceptional benign tumors representing 0,6% of the bladder tumors. They present as solitary unique lesions and can regress spontaneously as a result of fibrosclerosis suggesting a conservative approach wherever possible. They have a low potentional for recurrence. Asymptomatic hemangiomas do not require treatment. Case report: We report a case of a 52-year-old woman who was admitted to our hospital with the chief complaint of macroscopic haematuria, accompanied by lower abdominal pain. Cytoscopy revealed a red sessile tumor 1,2cm in diameter at the internal urethral orifice. Transurethral resection of the tumor was performed. The specimen was pathologically diagnosed as cavernous hemangioma of the urinary bladder. The typical microscopical feature was the presence of dilated and congested vascular spaces in the lamina propria of the mucosa of the urinary bladder. The patient was treated with biopsy and fulguration and she developed no recurrence 2 years after. Conclusions: Hemangioma of the urinary bladder is rare. The diagnosis is made histologically. Hemangiomas are classified into cavernous, capillary or arteriovenous types based on conventional criteria from other sites. A low incidence of recurrence is seen and the patients have a favorable outcome. Biopsy and fulguration are effective for hemangioma of the bladder when the lesion is small.

PP3-63 TERTIARY GLEASON PATTERN IN RADICAL PROSTATECTOMY SPECIMENS Arsenal Sezgin, Hakan Postaci, Guliz Ozkok Izmir Training and Searching Hospital, Department of Pathology, Izmir, Turkey BACKGROUND: The Gleason system is one of the most widely used grading system for prostatic adenocarcinoma. The Gleason score consisting of the sum of the grades of the primary (predominant) and the secondary (second most prevalent) grades, is an important prognostic parameter for prostatic carcinoma. Although the Gleason grading system does not account for the existence of a tertiary (third most prevalent) pattern, more than two predominant Gleason patterns can be present in one prostatectomy specimen. METHOD: Between 1998 and 2003, 51 patients underwent radical prostatectomy for clinically localized prostate cancer in our hospital. Prostatectomy specimens were examined retrospectively for Gleason scores including a tertiary grade and pathological stage including margin status, seminal vesicle invasion and extraprostatic extension. RESULTS: Overall, of the 51 patients, 18 (%35.3) were found to have a tertiary pattern. In previous studies concerning the prevalence of a tertiary Gleason pattern in prostate cancer, presence of more than two patterns were reported between %16-50. Of the 18 prostatectomy specimens with a tertiary pattern in this study, 11 presented tertiary pattern 5. The Gleason score was >= 7 in 16 (%89) of the prostatectomy specimens with a tertiary pattern. All of the prostatectomy specimens (%100) with a tertiary Gleason pattern had positive margins. Extraprostatic extension was observed in 8 (%44.4) and seminal vesicle invasion was observed in 6 (%33.3) of the 18 cases with a tertiary pattern. In our study, the prevalence of a tertiary pattern was found to be consistent with previous studies published before. CONCLUSION: Prostatic carcinoma shows multiplicity and hetereogenity in many cases and it is possible that a third and even a fourth most prevalent pattern can be identified. In recent years, the prevalence of a tertiary pattern in radical prostatectomy specimens has become more important. High grade tertiary patterns are found to be associated with a more advanced pathological stage. Although further studies are needed, the presence and percentage of a tertiary, particularly less differentiated Gleason pattern can be accepted as a potential new predictive factor for prostate cancer behaviour. PP3-64 IMMUNOHISTOCHEMICAL EXPRESSION OF RHOE IN BENIGN PROSTATE GLANDS AND PROSTATIC ADENOCARCINOMA Gabriel Matheu1, Joan Maria Benejam2, Priam Villalonga3 1 Servei d’Anatomia Patològica Fundación Hospital Manacor, and Institut Universitari d’Investigació en Ciències de la Salut (IUNICS). Illes Balears, Spain 2 Servei d’Urologia Fundación Hospital Manacor Illes Balears, Spain 3 Institut Universitari d’Investigació en Ciències de la Salut (IUNICS), and Departament de Biologia Fonamental, Universitat de les Illes Balears, Spain BACKGROUND: RhoGTPases are key regulators of the cellular cytoskeleton, but also regulate many other processes, including cell proliferation and transformation. Accordingly, there is increasing evidence for a role of Rho proteins in human cancer, and increased expression levels of Rho-family members are found in a variety of tumours. RhoE/Rnd3 is a member of the Rnd subfamily that, in contrast to other GTPases, do not hydrolize GTP and remain always in their GTP-bound active conformation. RhoE is a ubiquitously-expressed protein antagonizing RhoA function, promoting disassembly of actin stress fibres and focal adhesions. Recently, it has been shown that RhoE can also regulate cell proliferation and transformation. RhoE overexpression inhibits cyclin D1 expression leading to

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cell cycle arrest in fibroblasts and also in prostate cancer cells, in correlation with its lower expression levels in prostatic tumor samples relative to benign tissue, which have supported a tumor supressor role for RhoE in prostate cancer. METHOD: Formalinfixed and paraffin-embedded prostatic tissue sections obtained from 12 radical prostatectomy specimens were examined by immunohistochemical staining for RhoE (labelled streptavidinebiotine method, goat anti-Human RhoE antibody, Santa Cruz Biotech. Inc.®). The selected samples included benign prostatic glands (n=12) and adenocarcinoma scoring Gleason grade (G) 3 (n=10), G4 (n=9) and G5 (n=2). The staining index (SI) scoring system was defined by staining intensity on a scale of 0 to 4 multiplied by the estimated percentage of stained cells. Benign glandular epithelium was considered as positive internal control and graded 4+. RESULTS: All samples of benign prostatic glands showed positive citoplasmic staining for RhoE. RhoE expression was decreased or absent in malignant glands compared to benign epithelia. In addition, RhoE expression in prostatic adenocarcinoma was inversely correlated to the higher Gleason-grades of tumors. SI for G3=1.31; G4=0.41; G5=0. CONCLUSIONS: RhoE expression, detected by immunohistochemical analysis in prostatectomy specimens, is lower in neoplastic glands than in neighbouring benign epithelia. And RhoE expression levels inversely correlates to Gleason grade. The loss of expression of RhoE can support its tumor supressor role in prostate cancer, and may serve as a marker of prostate cancer progression. PP3-65 SERTOLI CELL MALIGNANT NEOPLASM IN A REDUCTIVE TESTIS. A CASE REPORT Irini Zinovieva, Ioannis Nesseris, Efthymia Balassi, Niki Agnantis Ioannina University Hospital, Greece AIM: To present a rare malignant testicular neoplasm with location in a reductive testis, which has been treated clinically as an extratesticular tumor. MATERIALS AND METHODS: A 25year-old male presented with a solid paratesticular mass measuring 4,5X3X1,8 cm. The intraoperative biopsy specimen, sent for a frozen section diagnosis, concerned a “paratesticular tumor” which was positive for malignancy. A tumor excision followed. The paraffin-embedded sections have been studied microscopically on hematoxylin-eosin preparations. An immunohistochemical study followed. RESULTS: The microscopic examination showed a neoplasm with a biphasic histological structure featuring tubular formations lined by high columnar, cuboidal and flat cells. Among them there were spindle cells. The neoplasm showed high pleomorphism, foci of necrosis and a high mitotic rate (30 mitoses per 20 high power fields). The immunohistochemical study showed positive for vimentin, MIC2, NSE, synaptophysin, antichymotrypsin, hCG, pancytokeratin and C-kit. The neoplasm was negative for EMA, CEA, ER, PLAP and chromogranin. CONCLUSION: A rare tumor can create diagnostic difficulties in intraoperative frozen section biopsy when is developed from testicular embryonic remnants and is mimicking a paratesticular neoplasm. PP3-67 HISTOMORPHOLOGICAL AND IMMUNOHISTOCHEMICAL EVALUATION OF POSTRADIOTHERAPY PROSTATE NEEDLE BIOPSIES Arzu Saglam1, Gokhan Ozyigit2, Fadil Aksoy2, Sevket Ruacan3, Arzu Sungur1 1 Hacettepe University Faculty of Medicine, Department of Pathology, Ankara, Turkey 2 Hacettepe University Faculty of Medicine, Department of Radiation Oncology, Ankara, Turkey 3 MESA Hospital, Department of Pathology, Ankara, Turkey

Introduction: Prostate needle biopsies have an important role in the follow-up of patients treated with radiotherapy, especially with regard to prognosis. However histomorphological interpretation of post-radiotherapy prostate needle biopsies is a challenging venture, mostly because practicing pathologists are not familiar with the histomorphological changes secondary to therapy. Methods: In this preliminary study, we evaluated the histomorphological changes induced by therapy in 46 of our patients treated with combined hormonal therapy and radiotherapy. All patients received the same therapy regimen and were rebiopsied after completion of radiotherapy. We used immunohistochemistry against Ki-67, vimentin, prostate specific antigen, chromogranine, p63 and high molecular weight cytokeratin in addition to H&E sections. Furthermore we compared the original diagnosis of these biopsy samples with the ones given after detailed morphological and immunohistochemical evaluation during this study. Results: The most prominent changes noted after therapy were as follows: acinar atrophy with decrease in acinus/stroms ratio (% 95.7’sinde), acinar distortion (%34.8), basal cell hyperplasia (%34.8), stromal fibrosis (%47.8), stromal edema (% 41.3) and vascular changes characterized by myointimal hyperplasia and luminal narrowing (%43.5). Nuclear enlargement (%100), nucleolar prominence (%80.4), bizzaar nuclei (%54.3) and cytoplasmic vacuolization (%93.5) was noted in the cells lining the prostate glands. Rare atypical cells in the stroma were seen to be basal cells positive for p63. Even though, originally 9 cases were reported to have residual adenocarcinoma and 3 reported as suspicious for adenocarcinoma, adenocarcinoma was confirmed in only 4 of these cases, 3 of which presented with distant metastasis during follow-up. The other cases were seen to display therapy induced changes. Most of these cases were diagnosed by practicing pathologists not experienced in prostate pathology and not familiar with the changes induced by therapy. Interestingly the morphological changes described above were not obvious in the neoplastic component of the 4 cases with residual foci of adenocarcinoma, even though they were noted in the adjacent nonneoplastic component of these cases. Conclusion: We concluded that H&E morphology is sufficient in diagnosing posttherapy needle biopsy specimens provided that the pathologist is experienced enough and familiar with the changes produced with therapy and that immunohistochemistry can aid diagnostic decision making in difficult cases. PP3-68 MULLERIANOSIS OF THE URINARY BLADDER: A DIAGNOSTIC PITFALL IN BLADDER BIOPSY SPECIMENS Sonuc Buyuk1, Ismet Basar2, Suleyman Ulucay2, Arkin Akalin3, Dilek Ertoy4, Burcu Albayrak5, Mehmet Muderiszade5 1 Department Of Pathology, Dr. Burhan Nalbantoglu State Hospital, Nicosia, North Cyprus 2 Department Of Urology,Dr. Burhan Nalbantoglu State Hospital, Nicosia, North Cyprus 3 Nicosia Diagnostic Centre, Nicosia, North Cyprus 4 Department Of Pathology, Faculty Of Medicine, Hacettepe University, Ankara,Turkey 5 Department Of Pahology, Dr. Burhan Nalbantoglu State Hospital, Nicosia, North Cyprus Müllerianosis is an entitiy characterized by the presence of different Müllerian-derived tissues in the wall of this organ. The terms endocervicosis and endosalpingiosis are used when only endocervical and tubal type glands are present respectively. When endoservical, tubal and endometrial glands are found in combination in the bladder it is defined as müllerianosis. It is a rare entitity and to our knowladge up to now six cases have been reported. We present a further case in a 37- year-old woman who had 2 caesarean sections before. The patient didn’t have any bladder symptoms unlike cases that have been reported earlier . She only complained from dysmenorrhea and metrorrhagia. A

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bladder mass was found by pelvic ultrasonography during routine control for diabetes mellitus. Ultrasonographic findings consisted of 16x19x30 mm solid hypoechoic lesion occupying the base and posterior wall of the bladder projecting into the lumen. A complete transurethral resection , including the deep muscular layer, yielding 10 chips was performed . Cystoscopic gross appearence was different from the usual transitional cell carcinoma or adenocarcinoma, and was rather displaying a cystic form. Once the resection started, the inner surface and consistency of the tissues were mimicking the gross appearence of adenocarcinoma ( of a solid tumour ). Histologically there were intermediate to large-sized, irregularly shaped , haphazardly arranged, endocervical type glands in the lamina propria and in the muscularis propria, some of which were cystically dilated. In some areas rupture of the glands and inflamatory respond were seen . Siderophages also were noted in lumens and in the stroma. In focal areas there was endometrial type glands which were typically surrounded by endometrial stroma. Well differantiated adenocarcinoma has been taken into concideration. Since there was no atypia and desmoplastic reaction ,the presence of endometrial type glands and its typical stroma and the metaplastic change of superficial urothelial epithelium into the endocervical type epithelium lead us to the correct diagnosis. Although our clinicopathological findings supported metaplastic origin, still the clinical history of prior caesarean section puts doubts to this. PP3-69 DISTRIBUTION PATTERNS OF NEURAL NETWORK ON SURFACE OF PROSTATE AND EXTENDING PATTERNS OF PERINEURAL INVASION OF PROSTATIC CANCER. A MORPHOLOGICAL STUDY USING 3-D RECONSTRUCTION METHOD Mareyuki Endoh1, Taiyi Jin2, Ryoji Chiba1, Noriyuki Iwama1, Hironobu Sasano2 1 Dept. of Pathology Sendai Kousei Hospital, Japan 2 Dept. of Pathology Tohoku University Hospital, Japan Background: Perineural invasion would be frequently observed prostatic acinar adenocarcinoma. Although, not only distribution patterns of the invasion but also its patterns of neural network on surface of prostate were not clarified completely. So as to elucidate the mechanism of perinaural invasion, we selected several cases of prostatic cancer with extensive perineural invasion. However the distribution patterns were threedimensional and complicated, thus we analyzed completely serial sections of prostatic cancer specimens with a computer assisted 3-D reconstruction system for this study. Materials and methods: Four surgical specimens of total prostatectomy, with cancers having extensive perineural invasion, were used. After formalin fixed and paraffin embedded, complete serial sections were mede from the specimens as 4 um thickness and H&E stained. Every four serial sections were used for strict tracing the pattern of nerve branch and carcinoma invasion, and the data were digitalized. The data were incorpotaded in a computer assisted 3D reconstruction system and 3-D images were constructed in all its aspects for morphological analysis in detail. Result: Each number of serial sections was 170, 262, 265 and 267 sheets and their thicknesses were 510, 786, 795 and 801 um, thus searched total thickness of the prostatic tissues reached 2,892um. The 3-D images in all its aspects were apparently indicating the distribution patterns of the neural networks and extending patterns of the carcinoma, as shown in the conclusion below. Conclusion: The 3-D images showed the neural networks made complicated distribution patterns with apparent ganglions. The nerves extended narrow branch toward prostatic parenchyma. Even if tiny perinaural invasion would be showed in prostatic parenchyma, the extension of cancer could be spreading widely for outside of the prostate through the neural network. However, the invasion hardly extended from perineural space into surrounded soft tissue. The finding might explained a hypothesis

that perineural invasion should not be a prognostic factor of prostatic cancer. PP3-70 DECLINING DIAGNOSTIC VALUE OF HIGH-GRADE PIN IN PROSTATIC BIOPSIES Jurg Vosbeck1, Seife Hailemariam2, Lukas Bubendorf1 1 Institute for Pathology, University of Basel, Switzerland 2 Cantonal Institute for Pathology, Liestal, Switzerland Background: The detection of isolated high-grade prostatic intraepithelial neoplasia (HG-PIN) in prostatic core biopsies has long been considered an important risk factor for a concomitant adenocarcinoma. Given an estimated risk of 30%, re-biopsy within 6 months has usually been recommended. However, the diagnostic importance of HG-PIN has recently been questioned. Here, we investigate the diagnostic relevance of isolated HG-PIN in the biopsy material submitted to our institutes of pathology during the past 10 years. Methods and results: 125 patients with a diagnosis of isolated HG-PIN were identified in our databank from 1995 to 2005. All histological slides were re evaluated by the authors according to standardised criteria, and compared to the result of re-biopsies. Criteria for HG-PIN were unequivocal morphology with nuclear crowding, nuclear atypia with hyperchromasia and prominent nucleoli, as well as subepithelial retraction. 39 cases (31%) did not meet our stringent review criteria for HG-PIN and were excluded from further investigations. So far, a re-biopsy was done in 40 of the remaining 85 patients. Two of these re-biopsies (5%) showed an adenocarcinoma, and 9 (23%) showed isolated HG-PIN. 29 rebiopsies (73%) were inconspicuous. Conclusion: The diagnostic importance of isolated HG-PIN in a contemporary PSA-screened population is low, questioning the necessity of a short-term rebiopsy. Molecular markers may help to better stratify the diagnostic importance of HG-PIN in the future. PP3-71 IMMUNOHISTOCHEMICAL STUDY OF COX-2 EXPRESSION IN PROSTATE CARCINOMA; RELATION TO APOPTOSIS AND ANGIOGENESIS Seyda Erdogan1, Suzan Zorludemir1, Derya Gumurdulu1, Gulfiliz Gonlusen1, Gulsah Seydaoglu2, Emine Bagir1, Bulent Soyupak3, Zuhtu Tansug3 1 Cukurova University, Medical Faculty, Department of Pathology, Turkey 2 Cukurova University, Medical Faculty, Department of Biostatistic, Turkey 3 Cukurova University, Medical Faculty, Department of Urology, Turkey Background: COX-2 play role in tumorigenesis of a variety of human malignancies by stimulating cell proliferation, inhibiting epithelial differentiation, inhibiting apoptosis, mediating immune suppression and increasing the production of mutagens. The objective of this study is to determine if there is relationship between COX-2, apoptosis and angiogenesis in prostate carcinomas. Material and Methods: Paraffin-embedded tissue specimens from 49 prostate adenocarcinoma were retrieved from the files of the pathology department of Cukurova University, Medical Faculty. COX-2, Bcl-2, VEGF and M30 were studied immunohistochemically. The results of staining for COX-2, Bcl-2 and VEGF were analyzed semiquantitatively by using an immunohistochemical scoring system (HSCORE) that combines the percentage of immunoreactive cells (quantity score) and an estimate of staining intensity (staining intensity score). Apoptosis was figured out by monoclonal antibody M30, which reacts with the product from the cleavage of cytokeratin 18. The number of M-30 positive cells per 1000 cells was expressed as apoptotic index. Results: The age of the patients ranged from 48 to 73 years old (average, 63.8±6.4). Twenty-one (42.8%) of the cases were Gleason score ” 6 and 28 (57.2%) were Gleason score •7. COX-

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2 expression was detected in 81.6% of cases. It was significantly correlated with Bcl-2 expression (r=0.49; p<0.0001). There was no correlation between COX-2 and VEGF expression (p>0.05). Gleason score was negatively correlated with M30 (r=-0.28; p=0.04). No significant relation between Gleason score, VEGF and Bcl-2 was observed. Negative correlation between COX-2 expression and mean survival was also observed (p=0.007). Conclusion: Our study indicates that COX-2 plays role at the carcinogenesis of prostate carcinoma through inhiting apoptosis rather than promoting angiogenesis. This result may offer new insights into the treatment strategies. In addition, it may be useful for the prediction of clinical outcome. PP3-72 PROGNOSTIC VALUE OF THE EXPRESSION OF ECADHERIN, ȕ CATENIN AND CD44 IN BLADDER CANCER Duygu Dusmez Apa1, Tuba Karabacak1, Murat Bozlu2 1 Mersin University Faculty of Medicine Department of Pathology Mersin, Turkey 2 Mersin University Faculty of Medicine Department of Urology Mersin ,Turkey Background: Loss of expression of adhesion molecules may contribute to cancer progression. Epithelial cadherin (E-cadherin) interacts with cytoskeletal proteins through the catenin complex. CD 44 is also a surface adhesion molecule. The present study was undertaken to investigate the alterations in the expression of Ecadherin, BETA catenin, and CD44 in urothelial carcinoma of the bladder, and their prognostic value. Methods: 54 patients with primary urothelial carcinoma of the bladder were included in this study. The median age was 64 (range: 29-84 years), with 49 men and 5 women. Tissue samples were obtained by radical cystectomy in patients with invasive tumours and transurethral resections in patients with superficial tumours. The histological grading was performed according to the WHO-ISUP criteria. Patients were grouped as low grade and high grade. The pathological staging was done according to the TNM classification. The tumours were grouped as superficial (Ta and T1) and invasive tumours (T2, T3, and T4). Among the 54 tumours, 24 were classified as low grade and 30 as high grade. 42 were classified as superficial tumours and 12 as invasive tumours. The proportion of immunostained cells was used as a criteria for the evaluation. Results: The staining showed a membrane pattern, except for the CD44 where staining was also found in the cytoplasm. Loss of membranous E-cadherin immunoreactivity was higher in invasive and high grade tumours, but this finding was not statistically significant. Loss of expression of BETA catenin was significantly associated with higher grade (p=0.001), but CD44 failed to show the same correlation. Both BETA catenin and CD 44 were not associated with invasiveness. Conclusion: Several studies have examined the role of E cadherin in urothelial carcinoma revealing clear associations of decreased expression, with high grade and advanced stage tumours. Our study confirms this data although not statistically significant. These findings suggest that the expression of E-cadherin and BETA catenin might be useful prognostic markers for the clinical assessment of bladder cancer. Whereas several studies have shown an altered expression pattern of CD44, some reports suggested that it has no association with clinico-pathological variables. Our findings correlated with the second group. This retrospective study with a limited number of cases does not allow definite conclusions. Large prospective clinico-pathological studies to validate the prognostic value of Ecadherin and BETA catenin immunoreactivity should be performed.

PP3-73 SHORT TERM HIGH DOSE CORTICOSTEROIDS INDUCE APOPTOSIS IN PROSTATIC EPITHELIUM Bahar Muezzinoglu1, Mohammad Sayeedudin2, Gustavo Ayala2, Thomas M. Wheeler2 1 Kocaeli University Medical School Department of Pathology Kocaeli Turkey 2 Department of Pathology, Baylor College of Medicine, Houston. TX, USA Background: Corticosteroids (CS) induce apoptosis in various human cells. However, published data quantifying apoptosis in prostate glandular epithelium following short term high-dose CS is lacking. Design: Prostate tissue from 36 organ donors who died from head trauma and given high-dose CS for cerebral edema were evaluated. The prostate was the last organ removed of those organs consented for donation. Most of the prostate was frozen but a mid-transverse section was immediately fixed in 10% neutral buffered formalin. Apoptosis was quantitatively scored on this H E whole-mount section per 10 hpf in 4000 total epithelial cells counted. Age-approximated, non-tumor bearing autopsy prostate tissue (n=47) with no recent CS treatment served as controls. Result: The mean age for donors and controls was 32 and 39, respectively. For donors, mean apoptotic index (AI) of peripheral zone (PZ) was 1.47% compared to 0.72% in transition zone (TZ) (p< 0.05). Zonal apoptotic indices were significantly higher than the control autopsy prostate PZ and TZ epithelium; however, there was no significant difference in ejaculatory duct and seminal vesicle epithelium (table). Two donors had incidental prostatic adenocarcinoma and the tumor cells had AI of 0.1% and 0.02 % Conclusion: Short term high dose CS induces apoptosis in prostate glandular epithelium both in the PZ and TZ, an almost 50 fold increase over controls in both of these zones. However, ejaculatory duct and seminal vesicle epithelium remained unchanged in terms of AI. PP3-74 VALUE OF HIGH GRADE T1 SUBSTAGING IN BLADDER CANCER Isabel Trias, Isabel Español, Noemi Vidal, Santiago Bucar Clinica Plato Fundacio Privada, Spain BACKGROUND Since 90’s there is in the literature a discussion on the value of substaging high grade T1 bladder cancer (HGT1). In our center we perform substaging according to Younes criteria since 1996 and we have demonstrate a statistically significant difference in progression for initial high grade T1b and T1c (T1b,c) urothelial carcinomas (deep invasive submucosa) vs T1a (lamina propia invasive carcinomas). From 2001 we treat T1b,c with a specific protocol that includes re-TUR after 1st BCG instilation and then a radical surgery if its shows T1c or more, or pursuit with BCG treatment if there is T1a, T1b, CIS or nontumoral tissue. Some of the arguments against the value of substaging is that many of the so called deep invasive submucosal carcinomas could be underdiagnosed T2’s. The aim of this study is to know if substaging detects real high risk neoplasias or simply reflects infrastaged (not detected T2) carcinomas. METHODS We have reviewed our reTUR performed during 2001-2006 for HGT1. We define reTUR as the procedure performed before 2 months after the initial TUR. We also reviewed 27 cases (T1b,c) looking for evidence of lymphovascular invasion with H&E. RESULTS. 53 HGT1:12T1a; 22T1b,c (9/22 -group A- followed the entire protocol and 13/22 -group B- didn’t); 15T1; 4TX, with 17/53 reTUR. 9 came from T1b,c group A, 1 fromT1b,c group B, 3 from TX , 2 from T1a and 2 from T1. Group A re-TUR (9/9) results were 1HGT1c (radical cystectomy showed pT0 pN1) ,1HGT1A , 7 non-tumoral tissue. For group B there was 1/13 re-TUR with HGT1C . For the remaining 7 re-TUR patiens we found 2HGT1 (1 from TX 1 from T1a) and 5 non-tumoral tissue (2 from TX, 1 from T1A, 2 from T1). There was no muscle infiltrating carcinomas in any of the re-TUR. 4/13 T1bc group B and 2/15

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T1’s patients progressed, none of T1bc group A, none of T1a and none of TX progressed. Vascular invasion was found in 6/27 cases, 2/27 were doubtful and 19/27 were without invasion . 3/6, 1/2, 2/19, progressed. CONCLUSIONS. There is no evidence to consider HGT1b,c as understaged T2. We think that deep invasive HGT1 have a high risk of progression and merits a particular treatment protocol. Deep invasive submucosa carcinomas have the capacity of lymphovascular invasion and lymph node metastases. PP3-75 THE INCIDENCE AND CLINICOPATHOLOGICAL SIGNIFICANCE OF MULTIFOCAL PROSTATE CANCER Deniz Filinte1, Ozdal Dillioglugil2, Bahar Muezzinoglu1 1 Kocaeli University Medical School Department of Pathology, Kocaeli, Turkey 2 Kocaeli University Medical School Department of Urology, Kocaeli Turkey Background: The presence of multiple foci of prostate cancer within the same gland is not an uncommon finding. Various studies reported incidence of multifocality in prostate cancer between 67-87%. There is an index tumor (measured by largest volume), presumed to be the driver of prognosis and one or more separate accessory tumors. Design: Ninety-four radical prostatectomies were performed between 2003 and 2006 and processed as whole specimens with all tumor foci mapped. The cases didnot have any preoperative theraphy. The tumors were further analyzed in terms of number of tumor foci. Chi square test was used for the correlations between multifocality and and patient age, preoperative PSA, extracapsular extension, surgical margin positivity, lymph node metastasis and seminal vesicle invasion. Cases operated between 2003 and 2005 had preoperative sextant core needle biopsy. For cases operated in 2006 preoperative transrectal biopsy had 12 cores. Results: As a whole, in 41.5% of cases (39 cases) prostate cancer was multifocal. For cases operated in 2006 this incidence was 67.7% The mean number of foci was 1.68. The age and the preoperative PSA value were not different in unifocal and multifocal tumors. Unifocal tumors had more frequent extracapsular extension (ECE) than the multifocal tumors.(p=0.038). Lymph node metastasis, seminal vesicle invasion and surgical margin positivity didnot have any significant difference in terms multifocality. Conclusion: In our cases, unifocal prostate tumors are more frequent than the multifocal tumors. With the standard 12 core biopsy protocol the incidence of multifocal tumors are increasing when compared to sextant biopsies. Unifocal tumors are at higher pathological stage than the multifocal tumors. This may be attributed to, assimilation of smaller volume cancers by the large volume index cancer at the time of diagnosis. PP3-76 EFFECT OF SOY ISOFLAVONE SUPPLEMENTATION ON PROSTATE TISSUE, MARKERS OF PROLIFERATION AND APOPTOSIS AND HISTOPATHOLOGICAL ENDPOINTS OF TUMOR GRADE, VOLUME AND STAGE Gokben Yildirim Kupesiz1, Wael Sakr2, Michael Cher3, Mousumi Banerjee4, Edson Pontes3, Howard Parnes5, Omer Kucuk6 1 Department of Pathology, Akdeniz University, Antalya, Turkey 2 Department of Pathology, Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, Michigan, USA 3 Department of Urology, Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, Michigan, USA 4 Department of Biostatistics, University of Michigan, Ann Arbor, Michigan USA 5 Division of Cancer Prevention, National Cancer Institute, Rockville, Maryland USA 6 Division of Hematology and Oncology, Barbara Ann Karmanos Cancer Institute, Wayne State University, Detroit, Michigan,USA

BACKGROUND Isoflavones are among the most promising potential anticarcinogenic compounds in soybeans. Epidemiological studies indicate that consumption of soybeancontaining diets is associated with a lower incidence of certain human cancers like prostate cancer. METHOD Men diagnosed with localized prostate cancer who have agreed to undergo radical prostatectomy, brachytherapy or cryotherapy were enrolled in the study (21 African American, 12 White, and 1 Asian). Subjects were given soy isoflavone capsules containing 150, 300 or 600 mg genistein (PTI G-2535) or placebo capsules. The study included a four-arm, double blind, placebo-controlled parallel group dose ranging trial: arm 4 -placebo; arm 3 -150 mg; arm 2 -300 mg; arm 1 -600 mg. Both biopsy and radical prostatectomy specimens belong to this eligible patient group were examined by using H&E and immunohistochemical biomarkers. Biomarkers of cell growth, bcl-2, MIB-1, cyclinD1, CDK5, CDK6; biomarkers of cell differentiation, apoptosis, and growth control, bax, p21, p53, Cx43 and biomarkers of angiogenesis, MVD, VEGF expression in tissues were evaluated by immunohistochemistry. The results were compared with the pathological stage and tumor volume on radical prostatectomy specimens. RESULTS No statistically significant change from pre to post treatment levels in any group and no statistically significant difference between the placebo and experimental groups in terms of change from pre to post levels for Cx43, MIB1, p21, bcl-2, bax, p53, cyclin D, CDK5, CDK6, MVD and VEGF were found. Statistical analysis revealed no significant differences between the groups except in margin positivity, which was much lower in group 1 compared to other groups. CONCLUSION Our results did not show any difference between our study groups with respect to markers of proliferation and apoptosis. This may be partly due to small sample size. PP3-77 THE IMPACT OF UNUSUAL HISTOMORPHOLOGICAL VARIANTS ON PATIENT OUTCOME WITH HIGH GRADE MUSCLE-INVASIVE UROTHELIAL BLADDER CARCINOMA: CLINICOPATHOLOGICAL STUDY OF 63 CASES B. Handan Ozdemir, Gulnur Guler, Aysel Colak Baskent University, Faculty of Medicine, Ankara, Turkey Our aim is to investigate the unusual histomorphological variants of high grade muscle-invasive urothelial cell carcinoma of the bladder and to assess its influence on prognosis. For this purpose we re-examine total 350 cases with urothelial bladder carcinoma. Of 350 cases 63 (18%) showed diverse morphologic features including micropapillary (10 cases), microcystic (8 cases), lymphoepithelioma-like carcinoma (3 cases), sarcomatoid carcinoma (eight cases, including pseudoangiosarcomatous type), osteoclast-rich undifferentiated carcinoma (4 cases), clear cell (5 cases), lipoid cell (5 cases), nested (3 cases), plasmacytoid/lymphoma-like variant (4 cases), rhabdoid cell (3 cases), signet ring cell (2 cases), small cell carcinoma (4 cases), inverted type (3 cases), and syncytiotrophoblastic giant cells (1 case). The extension of these non-conventional histomorphological features in tumor and the number of histological differentiation types in each tumor were examined. We found that the extension of unusual morphologic features with any proportion of conventional urothelial carcinoma had an unfavorable impact on survival time. Survival time of patients decreases significantly with increasing amount of nonconventional histological type in tumor. We also noticed that the increased number of differentiation types had an unfavorable influence on patient outcome. In conclusion we suggest that the assessment of the extension and the number of unusual histological differentiation types may be an important prognostic factor in high grade and muscle-invasive tumors.

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PP3-78 THREE DIFFERENT SIMULTANEOUS TUMORS IN ONE PATIENT; CLEAR CELL TYPE RENAL CELL CARCINOMA, LOW GRADE COLLECTING DUCTUS CARCINOMA AND MICROPAPILLARY UROTHELIAL CARCINOMA Ipek Isik Gonul1, Asli Cakir1, Sinan Sozen2, Omur Ataoglu1, Turgut Alkibay2 1 Gazi University School of Medicine, Department of Pathology, Turkey 2 Gazi University School of Medicine, Department of Urology, Ankara, Turkey Background: The coexistence of multiple and synchronous primary neoplasms in the genitourinary system has rarely been described in the literature. The case represented here was much more exciting since collecting ductus carcinoma has a low grade morphology,which is very rare. Method: Our case was a 57-yearold man who presented with prostatism for 2 months. Cystoscopy showed a mass in bladder, and abdominal CT revealed two masses in the middle segment and lower pole of the right kidney. With the preoperative diagnosis of high grade urothelial carcinoma, total cystoprostatectomy with urinary diversion and obturatory lymph node dissection was performed and 2 months later he underwent partial nephrectomy for the excision of renal tumors. Result: Grossly, 5x4x3 cm nodular lesion was found in the left lateral wall of the bladder. Microscopically, high grade infiltrative urothelial carcinoma with squamous differentiation and micropapillary growth pattern invaded the perivesical fatty tissue. Two metastatic lymph nodes were identified. One of the masses of the kidney had a pale gray, sponge-like cross section and it was 5.5 cm in its maximum diameter. Histology revealed a tubulocystic architecture characterized by dilated tubules lined by cuboidal to columnar cells that often have a hobnail pattern in a desmoplastic stroma. Tumor cells were positive for HMWK, vimentin, EMA and RCC. The lower pole kidney tumor was an obvious clear cell renal cell carcinoma with Fuhrman grade 2. EMA, vimentin and CD 10 immunoreactivity was detected but HMWK was negative. Conclusion: When synchronous tumors are diagnosed, standart sequential therapies may need to be reconsidered because of the possible problems concerning the effectivity of the treatment. Our patient is now being treated by a chemotherapy protocol against the metastatic carcinoma of the urinary bladder. Although we know that collecting ductus carcinoma is an agressive variant of renal carcinomas and must be treated seriously, our case was one of the very rare reported examples of “low grade” morphology with favorable prognosis. Therefore, it was not considered as our priority. PP3-79 INVESTIGATION OF CYP 1A1 AND GST M1 GENE POLYMORPHISMS AND P53 IMMUNHISTOCHEMICAL STAINING WITH CLINICAL AND PATHOLOGIC FEATURES IN PATIENT WITH TRANSITIONAL CELL BLADDER CARCINOMA Tulin Ozturk1, Haydar Durak1, Oguz Ozturk2, Turgay Isbir2, Canser Cakalir1 1 Istanbul University Cerrahpasa Faculty of Medicine Department of Pathology, Turkey 2 Istanbul University Institute of Experimental Medicine Department of Molecular Medicine, Turkey INTRODUCTION: Bladder cancer is the second most common malignancy of the genitourinary tract and the fourth most common type of neoplasm in men in many industrialized countries. Genetic status of the individuals and environmental interactions are thought to play an important role. Individual differences in cancer susceptibility may be mediated in part through polymorphic variability in the bioactivation and detoxification of carcinogens. The aim of this study was to identify the polymorphisms of the GST M1 and CYP 1A1 genes,

making correlation between those polymorphisms and the P53 expression as well as comparing with pathological clinical parameters with the patients with transitional cell carcinoma of the bladder. RESULTS: We obtained pathologically proven transitional cell carcinoma samples from 100 (19 female, 81 male: mean age 63.69±14.42 years) bladder cancer patients who underwent transurethral resection (TUR) or radical cystectomy between 2000 and December 2005 at Istanbul University Cerrahpasa Faculty of Medicine, Department of Pathology. Healthy individuals, 101 total; 33 female, 68 male; mean age 56.46±12.86 years were chosen as the control group.DNA was extracted from formalin – fixed, paraffin –embedded blocks of the patients. We used cut-off value for P53positivity of 10% of cells or more. There is no significant difference in gene polymorphisms between the patients and control group, however, CYP 1A1 heterozygote mutant genotype was less common in patient group. GST M1 null genotype is not different between patients and conrol groups but increasing risk for perineural and vascular invasion, multifocality, involvement of the pelvic lymph nodes, metastatic spread and death from the disease. and also invasive growth pattern is greater in the patients with GST M1 null genotype. These patients show higher p53 expression as well.There is a significant difference in p53 expression between CYP 1A1 homozygote wild and both mutant genotypes. CONCLUSION: In our study, although there was no significant difference between patient and control groups for GST M1 nullgenotype we found that the patients with GST M1 null genotype had poor outcomes than the others. P53 immunoreactivity was also statistically higher in the patients who have GST M1 null genotype. PP3-80 A RETROSPECTIVE STUDY ON BLADDER NEOPLASMS Gulistan Gumrukcu, Senay Tosun, Bilgen Sari, Selvinaz Ozkara, Pembegul Gunes, Murat Erkan Haydarpasa Numune Education and Research Hospital , Department of Pathology , Istanbul, Turkey In Haydarpasa Numune Hospital of Pathology Laboratuary , the bladder TUR and cytology cases which have been examined between 2003 August-2007 February monts were documented. 893 TUR ve 26 urine cytology , that belongs to 505 patients, which have been sent to our laboratuary between retrospectively reviewed. The cases were between 6-91 years old. There were 802 men and 117 women. 654 of all cases were reported malign and the other 265 were benign. The 52 cases of the group , which was reported malign, were papillary urothelial neoplasia- low malignant potential, 287 low grade carcinomas and 301 high grade carcinomas. One case of all the cases in that malign group was squamous cell carcinoma, 1 case high grade malign lenfoma infiltration, 1 case embryonal rhabdomyosarcoma, 2 cases carcinosarcomas of the bladder, 2 cases adenocarcinoma infiltration, 1 case dissemine insitu carcinoma and the other 2 cases were described as amyloidosis. According to Mostofi grade, 286 of the low grade carcinomas cases , were grade II and only 1 case was grade III. 116 of the high grade carcinomas cases reported as grade II, 185 as grade III. Lamina propria invasion in 26 cases has been diagnosed low grade carcinomas group. In 147 cases of the high grade carcinomas detrusor muscle invasion defined. 40 cases reported as inistu carcinoma determined. In 162 cases squamous differantiation was reported. The 3 of the benign lesions were inverted papillom, 3 flat hyperplasia, ,1 nephrogenic adenoma, 1 papillom and 12 cases were reported papillary urothelial hyperplasia. When 180 cases described as chronic specific and nonspecific chronic cystitis, the other cases defined as normal urothelial mucosa.

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PP3-81 SURVIVIN, INHIBIN AND p16INK4a EXPRESSION IN PROSTATIC ADENOCARCINOMAS Figen Ozturk1, Hulya Akgun1, Fatma Tokat1, Atilla Tatlisen2 1 Erciyes University School of Medicine, Department of Pathology, Kayseri,Turkey 2 Erciyes University School of Medicine, Department of Urology, Kayseri,Turkey BACKGROUND: Prostate carcinoma is one of the most commonly occurring malignancies in Western men and is a leading cause of carcinoma-related deaths. The aims of this study were to characterize and compare the expression patterns of survivin, inhibin and p16INK4a in primary prostate adenocarcinomas. METHODS: Immunohistochemical staining for survivin, inhibin and p16INK4a was carried out on archival specimens from 21 patients who underwent transurethral resection. Six of the 21 patients were low grade prostate carcinomas (final Gleason grade 2-6) and 15 were high grade prostate carcinomas (final Gleason grade 7-10). The percentage of cells that expressed survivin, inhibin and p16INK4a were classified qualitatively into 4 categories (0, +1,+2, and +3) based on intensity of staining and percentage of cells. Chi-square test and Fisher’s exact test were utilized for the analysis. RESULTS: The median patient age was 63 years (40-76). p16INK4a analysis showed positive staining in 83.3 % of the low grade and 100% of the high grade carcinomas specimens. Five patients (23.8%) were positive for inhibin and 7 patients (33.3%) were positive for surviving, all of which were high grade prostate carcinomas. A statistically significant difference was found among p16INK4a, and survivin and inhibin expression. p16INK4a expression was significantly higher in prostate carcinomas (95.2%). We compared p16INK4a immunostaining scores with surviving and inhibin expressions. Regarding the inhibin and survivin immunohistochemical expression, 25% (5/20) were concomitant p16INK4a and inhibin positive, 35% (7/20) were p16INK4a and survivin positive cases. CONCLUSION: In this study, we have demonstrated p16INK4a staining might be a diagnostic aid particularly in prostate carcinomas and survivin and inhibin expression might be helpful for differentiating particularly high grade carcinomas. Increased expression of p16INK4a may be involved in the carcinogenesis of the prostate and may be a potential therapeutic target. Additional studies are needed to confirm the potential value of p16INK4a, survivin and inhibin expression in prostate adenocarcinomas. PP3-82 EPIDEMIOLOGY OF TESTIS TUMOURS AFTER 50 YEARS OLD Camparo Philippe1, Comperat Eva2, Bertocchi Céline1, Richard Francois2, Capron Frédérique2, Houlagtte Alain1 1 HIA Val de Grace, Paris, France 2 Hopital Pitié Salpétrière, Paris, France Objective : In attempt to identify causes of testicular tumors among adult after 50 years old. Materials and methods : A retrospective study was made from patients treated in the military hospital Val de Grace and hospital la Pitié Salpétrière in Paris. Clinical data and diagnosis were obtained from pathologic reports. Populations were divided according age of onset : group 1 : post pubertal patients of less than 40 years old, group 2 : patients between 40 and 49 years old, and group 3 : patients of 50 or more years old. Statistical analyses were made with Epiinfo 6.04d. Results : Between 1994 and 2006, 392 testis tumors were treated in the military hospital Val de Grace (252) and la Pitié Salpétrière (140). Mean age of patient was 35 (median 31 extreme 16-85). 281 tumors (72%) occurred in group 1, 52 tumors (13,3%) were observed in group 2 and 59 (15%) in group 3. Germ cell tumors were the most common tumors observed representing 70,9%. Sex cord stromal tumors (SCT) represented 10% and other tumors 19,1%. Regarding age of onset, germ cell

tumors represented 77,23% in group 1, and 80,77% in group 2. Non-seminomatous germ cell tumors and seminomas were observed with the same frequency before 40 years old. Seminomas were the main germ cell tumor observed after 40 years old (57,69% vs 23,08% for non seminomatous germ cell tumors in group 2). In group 3, germ cell tumors represented only 32,2% cases (seminomas 23,7%, NSGT 8,5%). Sex cord stromal tumors represented 18,6% and other tumors 49,1%. Among these, lymphoma were the most common (10 cases, 17%). Tumors developed from mesothelia represented 6,8% (adenomatoid tumors 2 cases, malignant mesothelial tumor 2 cases) and spermatocytic seminomas 3,4% (2 cases). Five tumours were developed from testis adnexa (2 sarcomas and 3 benign tumors), 2 were testicular metastasis (one prostatic and one colonic cancer), 2 were benign epidermoïd cyst and 3 pseudotumoral lesion (2 orchiepididymitis and 1 amyloid pseudo tumor of the testis). Conclusion: Germ cell tumors are the most frequent tumor observed in adult testis but their incidence drop dramatically after 50 years old. At this age, sex cord stromal tumors and lymphomas are common and the use of lymphoid markers CD20 and CD79a, plus inhibin, Melan A and calretinin may be usefull in the diagnosis of a testicular tumor. Spermatocytic seminomas and adnexial tumors are very uncommon and one should consider the possibility of a metastatic origin in a tumor with atypical histologic patterns. PP3-83 PRIMARY MUCOSA-ASSOCIATED LYMPHOID TISSUE (MALT) LYMPHOMA OF THE URINARY BLADDER Ahmet Midi1, Cem Comunoglu2, Sinan Ekici3, Alper Eroglu3, Onder Peker1 1 Maltepe University, Department of Pathology 2 Oruc Pathology Laboratory 3 Maltepe University, Department of Urology , Istanbul, Turkey Background: Primary malignant lymphoma of the bladder is a rare tumor and account for less than 0.2% of all extranodal lymphomas. We here present a case of primary low-grade B- cell lymphoma of the MALT type of the urinary bladder. Case report: Fifty-six year old male patient presented with frequent urination, dysuria and periumbilical pain. Cystoscopic biopsies from bladder neck, right and left bladder wall, and base were taken. Histological findings revealed a dense lymphoid infiltrate. Widespread and dense mucosal lymphoepitelial lesions were present. The infiltrate mainly were composed of centrocyte-like cells and infrequent immunoblast/centroblasts. Scattered lymphoid follicles that had active residual germinal centers were seen. Immunohistochemically CD3, CD20, CD30, CD23, CD5, BCL-2, Cyclin D1, and Ki-67 were performed. CD3 was positive in 10% of the small cells, CD20 was positive in 90% of all cells, CD30 was negative, CD23 was positive at follicular dentritic cells in residual germinal centers, CD5 was positive in 10% of the small cells, BCL-2 was positive in 90% of all cells, Cyclin D1 was negative, and Ki-67 was positive in 20% of the neoplastic cells. With these histomorphological and immunuhistochemical findings, a diagnosis of extranodal marginal zone lymphoma (Malt lymphoma WHO-REAL), was given. Conclusion: Urinary bladder’s lymphomas are divided into three clinical groups: 1Primary lymphoma localized in the bladder, 2- Lymphoma presenting in the bladder as the first sign of disseminated disease (Nonlocalized lymphoma), 3- Secondary lymphoma. Our case was a primary lymphoma. Clinical and histopathological diagnosis of the MALT type lymphomas may be difficult. Immunohistochemical findings help us in recognition of these tumors in cases displaying lymphoepithelial lesions and a dense lymphocytic infiltration.

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PP3-84 MORPHOLOGICAL AND IMMUNOHISTOCHEMICAL CRITERIA OF PROGNOSIS IN RENAL CELL CARCINOMA Margarita Rybakova1, Salman Al-Shukhri2, Yulia Ponomareva2, Andrei Loukyanov2 1 Department of Pathology, St Petersburg State Medical University, St Petersburg, Russia 2 Department of Urology, St Petersburg State Medical University, St Petersburg, Russia Aim of the study. To estimate histopathological and immunohistochemical features of renal cell carcinoma with regard to clinical course and prognosis. Material and methods. Biopsy specimens from 47 patients (aged 35-75) with renal cell carcinoma were studied histologically and immunohistochemically. Grading was performed according to Fuhrman. Immunohistochemistry was done for Ki-67, p53, bcl-2, CD34, N-cadherin and bax. Clinical records were evaluated with regard to type of surgery (resection or nephrectomy), tumor size, relapse and metastases. 13 patients showed relapsing and/or metastatic disease during 6-18 months follow-up after surgery, in 34 - the disease was local and non-progressive. Statistical analysis included p values and correlations. Results. Nearly all cases with progressive disease showed marked histological atypia. In these cases the capsule was thin or absent, the tumor had 3d -4th nuclear grade, and was characterized by low expression of N-cadherin and high proliferation rate (assessed by Ki-67 immunostaining). No clear difference was seen in bcl-2 and bax expression between the groups. A common feature associated with progressive disease was abundance of sinusoidal vessels, and invasion of tumor cells into these vessels was frequently seen. Expression of p53 gene product showed strong positive correlation with tumor nuclear grade. Conclusion Renal cell carcinomas with marked histological atypia, 3d -4th nuclear grade, having thin or no capsule, with abundance of sinusoidal vessels, low expression of N-cadherin and high proliferation rate are associated with progressive disease and unfavorable prognosis. Size of the tumor had minor influence on clinical course and prognosis in our setting. PP3-85 p27 AND HIGH MOLECULAR WEIGHT CYTOKERATIN EXPRESSION IN PROSTATIC ATROPHY AND POSTATROPHIC HYPERPLASIA Cumhur Ibrahim Bassorgun, Betul Unal, Irem Hicran Ozbudak, Bahar Akkaya, Mehmet Akif Ciftcioglu Akdeniz University, School of Medicine, Department of Pathology, Antalya, Turkey Background: Longstanding chronic inflammation and atrophy of the prostatic glands have known as linked entities. Development of carcinoma in a background of chronic inflammation has shown recently in several organs. This association between inflammation and carcinoma has also been supposed as a potential mechanism regarding the prostatic carcinogenesis. Atrophy of the prostate is identified as a reduction in the volume of preexisting glands and stroma and can be divided into two major patterns, diffuse and focal. Recently, a theory was suggested as the atrophy of prostate glands and the subsequent proliferation of the glandular epithelium represent the initial lesion, which can develop into cancer. Prostatic epithelium is stained with cytokeratins and p27 immunohistochemically. p27 is a cyclin-dependent kinase inhibitor which decreases in majority of prostatic adenocarcinomas and high-grade prostatic intraepithelial hyperplasia. In this study, we evaluated the expression of p27 and high molecular weight cytokeratin (HMWC) in prostatic epithelium. Method: The study was performed in 15 prostate tissues with prostatic atrophy and postatrophic hyperplasia. Prostate tissue samples were stained with p27 and HMWC immunohistochemically. Percentage of nuclear

staining in epithelial cells with p27 was assessed semiquantitatively. T-test was used for statistical analyses. Results: All atrophic and post-atrophic hyperplastic epithelial cells were stained with HMWC discontinuedly. The average count of p27 positive cells per 1000 cells were found 67 in atrophic glands and post-atrophic hyperplastic glands, but 79 in normal prostatic glands. There were statistically significant difference between p27 values of atrophic, post-atrophic hyperplastic glands and normal prostatic glands (p=0.001). Conclusion: Downregulation of p27 is seen in prostatic epithelial cells of atrophy and post-atrophic hyperplastic glands. Considering the suppressor role of p27 on cell proliferation, downregulation of p27 may be a marker of carcinogenesis. Specific staining pattern of HMWC may be helpful to distinguish foci of atrophy from normal prostate glands. PP3-86 MIXED GERM CELL – SEX CORD STROMAL TUMOR OF THE TESTIS. A CASE REPORT. Milanka Mrþerla1, Mladen Ugljareviü2, Anto BlažAnoviü2, Božo Krušlin3, Margareta Ugljareviü4 1 Department of Pathology and Forensic Medicine, Clinical Hospital Osijek, Croatia 2 Department of Pathology, General Hospital Vukovar, Croatia 3 Ljudevit Jurak Universitiy Department of Pathology, Sestre Milosrdnice University Hospital Zagreb, Croatia 4 Clinical Hospital Osijek, Croatia Introduction: The mixed germ cell – sex cord stromal tumor of the testis is defined as neoplasm having neoplastic germ cells elements and neoplastic sex-cord stromal elements arranged in a diffuse pattern. Case report The case of a 45-year-old man with painless right sided testicular tumor is reported. Radiologic and laboratory investigations including serum alpha-fetoprotein (AFP) and human chorionic gonadotropin (HCG) showed no abnormalities. A right radical orchidectomy was performed. A histologic examination showed biphasic tumor which was composed of two different types of cells. The majority of the tumor consisted of quite unifom predominantly spindle cells, which were immunohistochemically positive for vimentin and S100 protein and focally for Į-inhibin. The second cell type were large cells with abundant clear cytoplasm arranged in a small clusters at the periphery of the tumor. These cells showed no reactivity for immunostains mentioned above neither for cytokeratines MNF 116 and 7, AFP, HCG and PLAP. Conclusion As any rare neoplasm the mixed germ cell – sex cord stromal tumor of the testis makes a huge diagnostic problem. Clinical and biological behavior of this type of testicular neoplasm is not yet completely discovered. PP3-87 PREDICITVE VALUE OF HISTOPATHOLOGICAL FEATURES OF HIGH GRADE PROSTATIC INTRAEPITHELIAL NEOPLASIA FOR PROSTATIC ADENOCARCINOMA Gupse Turan, Bahar Muezzinoglu Kocaeli University Medical School Department of Pathology, Kocaeli, TURKEY Background: High grade prostatic intraepithelial neoplasia (HGPIN) is considered precursor of prostate carcinoma. Its detection in biopsy specimens warrants repeat biopsy for concurrent or subsequent carcinoma. Aim: In this study we aimed to determine the incidence of cancer in prostate needle biopsies after an initial diagnosis of HGPIN and ascertain whether histologic and/or clinical findings can help to predict the increased risk of finding cancer on repeat biopsy. Methods: There were 49 patients diagnosed as isolated HGPIN between January 1996 and December 2005 and had at least one repeat biopsy. Each case was reviewed and the following histopathological features were evaluated on each needle biopsy: number and

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percentage of cores involved by HGPIN, number and percentage of HGPIN glands, percentage of gland involved by HGPIN, linear measurement and percentage of HGPIN glands in whole biopsy, pattern of HGPIN, feature of pattern (pure vs combined); nucleolar prominence, pleomorphism and mitotic activity in HGPIN glands. Clinical parameters including the age and preoperative PSA levels were recorded. The pathological results in repeat biopsies were divided into 3 groups: benign, HGPIN and cancer. Relationship of morphological findings of HGPIN and the clinical outcome were evaluated by chi-square and t test. Results: Prostate cancer was detected in %16.3 of the cases in repeat biopsies. Pure pattern of HGPIN and presence of pleomorphism in HGPIN were predictive to determine the cancer on repeat biopsy (p=0.013, p=0.02 respectively). The difference between groups for other morphological or clinical parameters were not significant (p>0.05). Conclusion:Our results indicate that presence of cellular pleomorphism and pure pattern of PIN in needle biopsies are the histological features requiring close follow up for unsampled synchronuous or metachronous prostate cancer. PP3-88 IS THE OUTER PERIPHERAL ZONE / SUB CAPSULAR TISSUE COMPARTMENT OF THE PROSTATE MORE SUSCEPTIBLE TO CARCINOGENESIS AND EARLY NEOPLASTIC TRANSFORMATION? A TOPOGRAPHIC MORPHOLOGIC AND TISSUE MICRO ARRAY IMMUNOHISTOCHEMICAL STUDY Gokben Yildirim Kupesiz1, Charles Butler2, Vashali Pansari2, Wael Sakr2 1 Department of Pathology, Akdeniz University, Antalya, Turkey 2 Harper University Hospital, Karmanos Cancer Institute & Wayne State University Hospital, Detroit, USA Background: The approach to prostate cancer detection using needle biopsies evolved in recent years to obtain more biopsy cores and to concentrate on sampling the outer and sub capsular areas of the gland including the apical/anterior compartment. Several studies have demonstrated that this modification from the traditional “sextant” method results in better detection of particularly smaller tumors. Our objectives were to 1-document the anatomical distribution of morphological lesions characteristic of early prostatic neoplasia including high grade prostatic intraepithelial neoplasia (HGPIN) and small foci of prostatic carcinoma (PCA) in two large series of radical prostatectomy specimens and of prostate gland examined in a post mortem setting. 2- To explore whether the outer glandular epithelium is likely to express markers associated with early changes of carcinogenesis. Method: Foci of PCA and HGPIN were mapped in a series of 83 consecutive, entirely embedded glands obtained during autopsies of young men (age 22-55) and in a series of 227 radical prostatectomy specimens (RPS) harboring small tumors (total tumor volume <=1 c). The distance of PCA and HGPIN foci from the outer surface of the gland was measured. Using the hematoxilin and eosin stained section, tissue microarrays (TMAs) were constructed from benign glandular epithelium of both subcapsular and inner peripheral zone locations from both the RPS and the autopsy prostates. The TMAs were stained with cell cycle regulator/proliferative markers (p27 and Ki-67) and with Glutathione S-Transferase 1 (GSTP1), Results: Of the 235 foci of PCA found in the 83 autopsy gland, 191 (81%) were within 5 mm of the outer surface of the gland. Similarly 176 of the 223(79%) foci of HGPIN in this cohort had the same distribution. For RPS, 112 (49%) of the small volume tumors were also within 5 mm of the outer surface. GSTP 1 showed less diffuse distribution and with lower intensity in the outer versus the inner compartment of the gland. Similar pattern was found with p27 immunoreactivity while the proliferation rates were low in both compartments but slightly higher in the most outer epithelium. Conclusion: Most small volume PCA, (both sub clinical and low stage clinically

detected), and most HGPIN are present in the outer subcapsular prostatic tissue. Our morphologic and TMAs findings indicate a higher vulnerability of the epithelium in the outer peripheral zone to early neoplastic transformation and are consistent with the recent biopsy sampling approaches. PP3-89 MORPHOLOGICAL FEATURES OF KIDNEY CYSTS OF III BOSNIAK CLASSIFICATION TYPE Pasechnik Dmitry Rostov Medical University, Russia Kidney cysts of III Bosniak classification type are the most complex problem in malignancy potential assessment and adequate treatment strategy selection. The aim of the research was to investigate morphological features of III type kidney cysts. 15 kidneys operated under following conditions were investigated: kidney cancer (5), multilocular cysts with possible malignant transformation (5), multicystic dysplasia of the kidney (3), benign cysts (2). There were investigated 10 males and 5 females. The median age was 54.8±2.35 years. All samples were of the following histological structure: clear cell renal cell carcinoma with cystic degeneration (4 cases), cystic renal cell carcinoma (6 cases), multicystic dysplasia of the kidney (3 cases), angiomyolipoma with hemorrhages and cystic degeneration (1 case). There was one case of synchronous multifocal tumor of the kidney: 3 angiomyolipomas were combined with cystic nephroma adult type. Cystic renal cell carcinomas are characterized by single cysts inner lining preferably of rounded or columnar cells with clear vacuolated cytoplasm, which are much in common with typical clear cell renal cell carcinoma. Similar cells were revealed in cystic cavities fluid under cytological investigation. All tumors were well-differentiated (Fuhrman nuclear grade -1) without invasive growth out of kidneys. Cystic nephroma was characterized by inner lining consists of cubical or hobnail cells monolayer with eosinophilic cytoplasm without signs of nuclear atypism and invasive growth. Thereby III type kidney cysts to Bosniak classification represent the heterogeneous group of tumorous and nonneoplastic diseases demanding preoperational and intraoperational morphologic assay to make a correct choice of type and extent of surgical treatment. PP3-90 eNOS AND iNOS EXPRESSION IN UROTHELIAL TUMORS OF THE URINARY BLADDER Ipek Isik Gonul1, Asli Cakir1, Nalan Akyurek1, Serhat Gurocak2, Sinan Sozen2, Turgut Alkibay2 1 Gazi University School of Medicine, Department of Pathology 2 Gazi University School of Medicine, Department of Urology, Ankara, Turkey Background: NO (nitric oxide) is one of the main factors responsible for the cytotoxic activity that macrophages exert against tumor cells. It also plays an important role in tumor growth and angiogenesis. NOS (nitric oxide synthase) has been found in various tumor types suggesting that NO may be produced in tumor tissue. The purpose of this study was to analyze the expressions of endothelial (eNOS) and inducible (iNOS) isoforms of NOS in the urothelial tumors of different grade and stage and to compare it with clinicopathological findings. Method: The medical records of 87 patients with primary urothelial carcinomas were reviewed. Grade according to WHO 2004 classification, pathological stage, invasion patterns of invasive tumors (nodular, trabecular, infiltrative, micropapillary) and stromal response types (edema and congestion, desmoplasia, fibroblastic proliferation, inflammatory response) were assessed by light microscopy. Paraffin embedded tumor sections were stained immunohistologically with eNOS and iNOS antisera and semiquantitative evaluations were performed. Association of eNOS and iNOS immunoreactivity with tumor grade, stage, invasion patterns and stromal response was examined with their

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prognostic significance in predicting tumor recurrence. Statistically p values of less than 0.05 was considered statistically significant. Results: Both iNOS and eNOS expressions in tumor cells were heterogeneous, ranging from diffuse but weak expression to focal but severe staining. 73 % and 88% of tumors showed iNOS and eNOS expressions respectively. eNOS was also identified in endothelial cells. Stromal inflammatory cells showed varying degrees of iNOS expression but it was occasional. Neither eNOS nor iNOS immunoreactivity did correlate with tumor grade, stage or invasion patterns (p>0.05). On the other hand, tumor grade and stage were found to be statistically correlated with the invasion patterns of tumors (p<0.05). Conclusion: Although biological role of NO in urothelial carcinomas is still unclear, this study demonstrated that both iNOS and eNOS were being expressed in urothelial tumors. Our results were inconsistent with the literature in that we observed eNOS expression in tumor cells together with the endothelial cells of the capillaries. A clear correlation with tumor grade, stage and recurrence rate was not identified. PP3-91 EXPRESSION OF CYCLIN D1, p16 AND Ki67 IN PROSTATE CARCINOMA Chariklia Kouvidou1, Grigoris Karpenisiotis2, Dimitrios Anagnostakis1, Haralambos Zorzos1, Evgenia Lianou1, Dimitrios Karanastasis2, Konstantinos Stravodemos3, Kitty Pavlaki4 1 Department of Pathology, ELPIS Hospital, Athens, Greece 2 Department of Urology, ELPIS Hospital, Athens, Greece 3 Department of Urology, LAIKO Hospital, Athens, Greece 4 Deprtment of Pathology, Med School, University of Athens, Greece Background: Alterations in cell cycle regulators and subsequent deregulation of the G1-S transition may cause uncontrolled cell cycle progression and may implicate in the development and progression of prostate cancer. The aim of this study was to investigate the expression of cyclin D1, p16 and Ki67 in prostate carcinoma and to correlate them with clinicopathological parameters. Materials and methods: Formalin-fixed paraffinembedded tissue from 50 cases of radical prostatectomy was stained by immunohistochemistry for cyclin D1, p16 and Ki67. The anti-cyclin D1 (Neomarkers), the anti-p16 (NeoMarkers) and the anti-Ki67 antibody (MIB-1, Dako) monoclonal antibodies were used at dilution 1:30, 1:50 and 1:30, respectively. A step of microwave heating in a solution of sodium citrate was performed prior to incubation with the antibodies. Associations between p16 (<20% vs. [GTEQ]20%), Gleason’s score (<7 vs. [GTEQ]7) and cyclin D1 (<10% vs. [GTEQ]10%), were evaluated using Fisher’s exact test, while for Ki67 (median 8%, range 2%-30%) with all the above, Wilcoxon was adopted. Results: Cyclin D1 positive nuclear staining was found in 12/50 (24%) prostate carcinomas (14% of low and 86% of high grade). Benign prostate acini showed focal scattered cyclin D1 positivity of the basal and secretory epithelial cells. P16 positive nuclear and cytoplasmic staining was observed in 23/50 (31% of low and 69% of high grade). Positive staining was not found in benign prostate or hyperplastic acini. Ki67> median was found in 36/50 (40% of low and 60% of high grade). Cyclin D1 expression showed a statistically significantly correlation with higher Gleason’s score (p=0.04) and advanced tumour’s stage (p=0.03).There was a tendency for a significant relationship between p16 expression and Gleason’s score (p-value=0.13) and a statistically significantly correlation between p16 and stage (p=0.0003). A trend for an inverse relationship between cyclin D1 and p16 was detected (p=0.09). A trend for a parallel expression between cyclin D1 and Ki67 was found. Conclusions: Cyclin D1 functions mainly in promoting cell proliferation and seems to play a role in the pathogenesis of prostate carcinoma. Increased p16 expression during tumour progression suggests that the senescence pathways are still intact in large number of prostate carcinomas.

PP3-92 FAS LIGAND EXPRESSION IN PROSTATE ADENOCARCINOMAS AND ITS RELATIONSHIP WITH HISTOPATHOLOGICAL FEATURES Sema Bircan1, Korkut Bozkurt1, Nilgun Kapucuoglu1, Alim Kosar2 1 Suleyman Demirel University School of Medicine, Department of Pathology, Isparta, Turkey 2 Suleyman Demirel University School of Medicine, Department of Urology, Isparta, Turkey Background: Fas ligand (FasL) is a type II transmembrane tumor necrosis factor family protein, known to trigger apoptosis in cells that bear the FasL receptor, Fas. The aim of the study was to examine the expression of FasL in benign prostatic epithelium (BP) and prostate carcinoma (PC) and to investigate its relationship with histopathological features in PCs. Method: The study included 51 primary PCs (22 radical prostatectomy and 29 transurethral resection) and 28 BP specimens. All slides were reevaluated histopathologically. In all cases Gleason grading and scoring, and in radical prostatectomy cases, extraprostatic extension, vascular and perineural invasion, high grade prostatic intraepithelial lesion (HGPIN), surgical margin status were recorded. Using tissue microarray slides, FasL examination was performed immunohistochemically. The staining was evaluated semiquantitatively as percentage of the stained cells as 1+, focal (<10%); 2+, moderate (10-50%); 3+, diffuse (>50%), and staining intensity was considered negative (0), mild (+), moderate (++), strong (+++). Results: Cytoplasmic and focal membranous FasL immunoreactivity was observed in 82.1% of BP (23/28) and 94.1% of PCs (48/51). The percentage of stained cells in PCs were classified as 10 (19.6%) focal, 10 (19.6%) moderate and 28 (54.9%) diffuse, and in BP cases, 8 (28.6%) focal, 10 (35.7%) moderate and 5 (17.9%) diffuse, respectively. The percentage of the stained cells with FasL in PCs was significantly higher compared with that in BP tissues (p=0.003). The PC cases with higher Gleason scores showed significantly much higher positively stained cells compared with that in lower ones (p=0.045). The intensity of FasL staining was also significantly different between PC and BP specimens (p=0.005). Mild (+) and moderate (++) staining was seen in 15 (53.6%) and 8 (28.6%) of BP specimens, respectively. Conversely, 20 (39.2%), 18 (35.3%) and 10 (19.6%) PC cases showed mild (+), moderate (++) and strong (+++) staining, respectively. In radical prostatectomy cases, the percentage of FasL staining was also significantly higher in cases with higher Gleason score and HGPIN compared with that in cases with lower Gleason score and without HGPIN (p=0.022, p=0.024, respectively). Conclusions: The increased FasL expression and its association with higher Gleason scores in prostatic adenocarcinomas suggest that FasL may be important biologic marker in the growth and progression of prostatic cancer and potential target of therapeutic intervention. PP3-93 PATHOLOGIC CHARACTERISTICS OF PROSTATE CANCER IN AFRICAN AMERICAN AND CAUCASIAN MEN DIAGNOSED AND TREATED AT OR BELOW 50 YEARS OF AGE: RACIAL DIFFERENCES ARE MOST EVIDENT IN THE YOUNGER AGE BRACKET Gokben Yildirim Kupesiz1, Isaac Powell2, David Grignon3, Wael Sakr3 1 Department of Pathology, Akdeniz University, Antalya, Turkey 2 Department of Urology, Harper University Hospital, Karmanos Cancer Institute & Wayne State University Hospital, Detroit, MI, USA 3 Department of Pathology, Harper University Hospital, Karmanos Cancer Institute & Wayne State University Hospital, Detroit, MI, USA Background: Data from Wayne State University suggest that the racial discrepancies in the incidence and outcome of prostate cancer between African American (AA) and Caucasian (C), men

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maybe more pronounced within the younger patient groups. The objective of this study was to compare pathological findings in both the diagnostic needle biopsy (NB) and the radical prostatectomy specimens (RPS) for the subset of men diagnosed, and the overlapping but no identical subset, who were treated by surgery at our institution. Design: For the NB subset, diagnostic serum PSA, number of cores involved by tumor and Gleason score for each involved core were recorded. For the RP subset, pathological stage, Gleason score, calculated tumor volume and extensiveness of high grade PIN were documented. Statistical analysis of the findings comparing pathological parameters between the two races for each subset was performed. Result: For the biopsy subset, 116 AA and 81 C patients were included in the study. The mean and median age for both groups were identical. Mean and median diagnostic PSA for AA were 7.4 and 5.5 ng/ml compared to 7 and 7:00 and 5.6 ng/ml for C [p< 0.05 for both values]. Forty five of the 116 (39%) AA patients, had three or more core involved by carcinoma compared to 19 of 18 (23%) C patients [p<0.05]. Nine Ten of 116 (8%) and 4 of 81 (5%) AA and C patients respectively had a biopsy Gleason score of 8 [p<0.06]. The RP cohort, included a nearly identical number of AA and C, (82 and 83 patients respectively). Organ confined (pT2), disease was found in 52 of 83 (63%) AA vs 44 of 82 (53%) C, [p<0.05]. While Gleason score distribution was similar, slight difference was found in the percentage of patients having Gleason pattern 4 or higher as their primary pattern (17% AA, 13 % C, [p= NS]). Larger tumors of 4 cc or greater were found in 31% of AA compared to 19% of C patients, [p<0.05]. Extensive HGPIN with diffuse involvement of the gland was evident in 52% vs 22% AA and C men respectively, [p<0.0005]. Conclusion: Our data suggest that the difference in pathological parameters indicative of a more aggressive prostate cancer appear to be sharper within the younger cohort of AA and C patients diagnosed with the disease. These data may indicate a potentially different pathogenesis of the disease in the younger African American population. PP3-94 POTENTIAL DIFFERENCES IN THE CLINICOPATHOLOGICAL PROFILE OF FAMILIAL/HEREDITARY AND SPORADIC PROSTATE CANCER Gokben Yildirim Kupesiz1, Isaac Powell2, David Grignon3, Wael Sakr3 1 Department of Pathology, Akdeniz University, Antalya, Turkey 2 Department of Urology, Harper University Hospital, Karmanos Cancer Institute & Wayne State University Hospital, Detroit, MI, USA 3 Department of Pathology, Harper University Hospital, Karmanos Cancer Institute & Wayne State University Hospital, Detroit, MI, USA Background: There is increasing evidence implicating family history as a distinct risk factor associated with the development and possibly the aggressiveness of prostate cancer. Recent epidemiologic studies suggest a stronger familial aggregation for prostate cancer than that associated with colon or breast cancer, two malignancies with well recognized familial components. The objective of this study was to compare the clinicopathological characteristics of the patients with family history to that of the remaining radical prostatectomy patients. METHOD: Between1991 and 1997, the 1,321 patients treated by radical prostatectomy for clinically localized prostate cancer were considered for the study. The clinical history /record of our database indicated that 1074 (81.3%) of those patients were considered as having “sporadic” prostate cancer with no familial links while 247 (18.6%) met the criteria for familial/ hereditary distribution of the disease, (information self reported by patients). Of the 247, 120 were Caucasians, 114 African Americans and 13 belonged to other ethnic groups. Patients with affected first degree relative (father) or second degree relative (brother “s”)

were used for this study. Results: The median preoperative PSA was 8.0, 7.9, 7.7 ng/ml in control group, father family history and brother history groups respectively. [p=NS], while the mean age at surgery was significantly younger for men with a history of a father having PCA vs the control 57 and n65 years respectively [p<0.05]. The percentages of patients with organ confined diseases were 40.4%, 47.1%) and 43.6% for patients with history of a father, a brother or no family member having PCA respectively. The corresponding figures for patient with extraprostatic extension were 18%, 16.9 % and 15 % respectively. Approximately one third 29.7% of patients with history of a father having PCA had Gleason Score 6 compared to 18.8% for patients with history of a brother having PCA and to 22.8 % for the control group. There were no differences among patients with Gleason Score 7 or higher. Median tumor volume was 3.33, 4.14 and 3.42 for patients with a history of a father, brother or no family member with PCA respectively. Conclusion A subset of patients with family history (those who have a first degree relative; father), with PCA had an earlier age of onset. The several parameters compared otherwise between the sporadic and the familial cases were not significantly different. PP3-95 PRESERVATION OF ANTIGENICITY IN NECROTIC POST-CHEMOTHERAPY CHORIOCARCINOMA Enric Condom Mundó1, August Vidal Bel1, Fina Climent Esteller1, Xavier Garcia Del Muro2, Josep Maria Piulats Rodriguez2, Josep Ramon Germà Lluch2, Eladio Franco Miranda3 1 Servei d'Anatomia Patològica. IDIBELL. Hospital Universitari de Bellvitge. L'Hospitalet de Llobregat. Barcelona, Spain 2 Servei d'Oncologia Mèdica. IDIBELL. Institut Català d'Oncologia. L'Hospitalet de Llobregat. Barcelona, Spain 3 Servei d'Urologia. IDIBELL. Hospital Universitari de Bellvitge. L'Hospitalet de Llobregat. Barcelona, Spain Introduction. Immunohistochemistry is occasionaly used to demonstrate cell lineage in spontaneously necrotic tumors, mainly carcinomas and lymphomas. Antigen preservation in necrotic post-chemotherapy germ cell tumors of the testis has not been reported. Objective. To test the immunoreactivity of necrotic metastates of testicular germ cell tumors. MATERIAL AND METHODS. Immunostaining for Cam5.2, EMA, CD10, ĮFP, PLAP and HCG on formalin fixed paraffin embedded tissue from resected residual metastatic masses (retroperitoneal lymph nodes: 9 specimens; lung: 6; liver: 2: mediastinal lymph nodes: 1) in patients treated with chemotherapy for germ cell tumors of the testis. Results. In the specimens from 12 patients necrotic tumor immunostaining was retained for Cam5.2, CD10 and HCG. The stained cells were mainly shadow syncytiotrophoblastic giant cells. The time from completion of chemotherapy to the resection of the residual tumors ranged from three to ten weeks. Postchemotherapy HCG serum levels were normal in all cases (prechemotherapy levels were elevated in each case). PLAP, EMA, ĮFP and CD30 did not stain the necrotic tumor tissues. CONCLUSION. Necrotic post-chemotherapy germ cell tumors of the testis may retain immunoreactivity for Cam5.2, CD10 and HCG, thus allowing a positive diagnosis of necrotic choriocarcinoma. PP3-96 EXPRESSION OF MUCINS AND CD10 IN ADENOCARCINOMA OF THE URINARY BLADDER Ljubinka Jankovic-Velickovic1, Vukica Katic1, Jasmina Gligorijevic1, Vesna Zivkovic1, Biljana Djordjevic1, Goran Marjanovic2 1 Institute of Pathology Nis, Nis, Serbia 2 Clinic of Hemathology and Clinical Immunology Nis, Serbia Introduction: Mucins (MUCs) are high molecular weight membrane glicoproteins. Expression of MUC 1, 2, 5AC, and 6 genes may change during malignant transformation of epithelial

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tissues according to characteristic pattern. CD10/neutral endopeptidase (NEP) is a membrane-bound zinc metallopeptidase which is expressed in urothelial neoplasms and normal urothelium. The aim of this study was to evaluate the expression of MUC1, MUC2, MUC5AC, MUC6 and CD10 in order to determine immunoprofile of the primary adenocarcinoma of the urinary bladder. Methods: Research was performed in 3 patients with vesical adenocarcinoma, at the Institute of Pathology Clinical Centre Nis. The specimens were fixed in 10% formaldehyde and embedded in paraffin. The sections were stained with HE. We detected mucin by immunohistochemical staining using the following monoclonal antibodies from Novocastra, Newcastle, UK: MUC1, MUC2, MUC5AC, MUC6 and CD10. As a control we used the normal surrounding urothelium. Results: All of the vesical adenocarcinoma displayed expression of MUC 1 (luminal membrane staining) and MUC 2 (cytoplasmic staining); MUC 6 in 2/3 cases, while MUC5AC was not detected. Expression of CD10 was present in one case. In the normal urothelium MUC1 was limited predominantly to the apical membranes of the umbrella cell layer, while CD10 was found to be expressed weakly or reaction was negative in the apical portion of the cytoplasm along the luminal surface. Conclusions: The mucin expression profile in adenocarcinoma of the urinary bladder allows the identification of MUC 1 together with intestinal mucin (MUC 2), and in certain cases MUC6. Expression of CD10 is indicative for absorptive-cell differentiation in vesical adenocarcinoma. PP3-97 METANEPHRIC ADENOMA: A RARE BENIGN TUMOUR OF KIDNEY Guldal Yilmaz1, Eda Akpinar1, Berkan Resorlu2, Yasar Beduk2, Ayse Sertcelik1 1 Ankara University, School of Medicine, Department of Pathology, Turkey 2 Ankara University, School of Medicine, Department of Urology, Ankara, Turkey Introduction: Metanephric adenoma is a rare epithelial tumour composed of small, uniform, embrionic-appearing cells. It occurs in children and adults with female predominance. Approximately 50% of these tumours are incidental findings and they are categorized as benign metanephric tumours of kidney. Case Report: A 31-year-old female patient suffering from abdominal pain underwent ultrasonographic examination and a right renal mass was detected. Computerized tomography revealed a cystic renal mass located in the medial portion of upper pole of right kidney, which measured 3.5x3 cm in dimension. The laboratory findings, including blood counts and biochemical data, were in normal range. With the presumption of renal carcinoma an operation was planned and she underwent partial nephrectomy. Pathological examination, macroscopically, revealed a centrally cystic mass of 3.5x3x3 cm in dimention, with central haemorrhage and cut surface of grayish-tan in colour. Microscopically, a highly cellular tumour composed of tightly packed small, uniform, round tubular and acinar structures, encapsulated with thick fibrous tissue was seen. The cells were monotonous, with small uniform round to oval nuclei with delicate chromatin and inconspicious nucleoli. The cytoplasms were pale and scant. There was loose oedematous and focally hyalinized stroma. At the periphery of the tumour, there were many psammoma bodies. No mitotic figures were seen. Immunohistochemically, the tumour cells show diffuse and strong nuclear staining for WT-1, and cytoplasmic staining for vimentin and high molecular weight keratin. Also, moderate CD57 staining was detected in most tumoural cell cytoplasms. EMA, CK7 and low molecular weight keratin showed only focal positive staining. CD10 and RCC were negative. Conclusion: Metanephric adenoma is an uncommon, morphologically distinct tumour type, with characteristic histopathological features. Despite its size, it is benign and should be distinguished

particularly from papillary renal cell carcinoma type 1 and Wilms’ tumour. These unique features of metanephric adenoma should be pathologically and clinically recognized because of its invariably benign course. PP3-98 CYTOKERATIN 7 AND ANTIMITOCHONDRIAL ANTIBODY EXPRESSIONS IN DIFFERENTIAL DIAGNOSES OF RENAL EPITHELIAL NEOPLASMS WITH EOSINOPHILIC CYTOPLASM Ayhan Ozcan, Yildirim Karslioglu, Armagan Gunal, Bulent Kurt, Omer Gunhan Department of Pathology, Gulhane Military Medical Academy and School of Medicine, Ankara, Turkey BACKGROUND: The distinction between epithelial renal tumors with eosinophilic cytoplasm, namely, chromophobe renal cell carcinoma (ChRCC), the eosinophilic (granular) variant of clear cell (conventional) renal cell carcinoma (CRCC), and oncocytoma may remain questionable in some cases because of overlapping morphologic features. We evaluated cytokeratin 7 (CK 7) and antimitochondrial antibody (AMA) expressions to determine their potential value in distinguishing these tumors. METHOD: In this study, 22 renal tumors were included. They consisted of 11 ChRCCs, nine oncocytomas and two eosinophilic variant of CRCCs. Immunohistochemical study was performed on formalin-fixed, paraffin-embedded tumor tissue samples of these cases for CK 7 and AMA using streptavidin-biotin method. RESULTS: CK 7 staining: Seven cases of 11 ChRCCs showed strong cytoplasmic immunoreactivity with conspicuous peripheral accentuation. Among the nine oncocytomas, one case was positive, three cases were completely negative and five cases showed only focal staining in less than 5% of the tumor cell population. For two eosinophilic variant of CRCCs, only one of them was immunoreactive with CK 7. AMA staining: Five of 11 ChRCCs showed diffuse cytoplasmic coarse granular immunostaining with peripheral accentuation. Eight of nine oncocytomas demonstrated with diffuse cytoplasmic, but fine granular staining, and lastly, in one of the two eosinophilic variant of CRCCs, diffuse cytoplasmic and haphazardly distributed coarse granular immunoreactivity was observed. Combined interpretation of CK 7 and AMA stainings: Five of 11 ChRCCs showed combined CK 7 (+) and AMA (+) with diffuse coarse granular with peripheral accentuation. Eight of nine oncocytomas were CK 7 (-) and AMA (+). One of the eosinophilic variant of CRCCs was also CK 7 (-) and AMA (+) like in most of the oncocytomas, but the staining pattern of AMA was randomly distributed coarse granular, rather than fine granular which was observed in oncocytomas. CONCLUSION: When we reviewed all cases retrospectively with these immunohistochemistry results, we concluded that three of the ChRCCs were needed to be diagnosed as oncocytomas and two of them were eosinophilic variant of CRCCs. Similarly, one eosinophilic variant of CRCC was actually a ChRCC according to the CK 7 / AMA expression patterns. Lastly, only one of the nine oncocytomas was immunohistochemically consistent with the ChRCC. In conclusion, these results suggested that using CK 7 and AMA together may be a discriminative diagnostic tool in the differential diagnoses of renal epithelial tumors with eosinophilic cytoplasm, in addition to distinctive immunostaining patterns of AMA expresion PP3-99 ONCOCYTIC PAPILLARY RENAL CELL CARCINOMA O El Lamine, Nadia Kourda, A Bouzouita, M Sfaxi, R Noomene, B Tayahi, J Kourda, M Ferchiou, M Chebil, Sarrah Baltagi Ben Jilani, R Zermani Charle Nicolle’s Hospital Department of Pathology, Tunisia Introduction: Papillary renal cell carcinoma (RCC) is subclassified in type 1 displaying cells with scanty pale, cytoplasm arranged in a single layer and in type 2 showing

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pseudostratified cells with eosinophilic cytoplasm. However, the existence of more variants of papillary RCC may be inferred by the recognition of few cases with different morphological features. Materials and methods: We report a series of 3 oncocytic renal papillary tumors with the aim of determining their clinicopathologic features. Results: Two patients were male and 1 female (median age, 66 years). All our patients were symptomatic (3/3) flank pain (1/3), hematuria (2/3). All of them were treated by radical nephrectomy. On macroscopic exam, Tumors (median size, 5,5 cm) were intrarenal , unifocal and well limited, with no extrarenal extension. Histologically, they consisted of thin, nonfibrotic papillae lined by a single layer of oncocytic cells, with finely granular eosinophilic cytoplasm and round regular nucleus exhibiting central nucleolus (Fuhrman grade 2). All cases were positive keratin 7, vimentin, racemase and ckit positivity in one case. Then immunohistochemical profiles and the pathologic features argued against the diagnosis of oncocytoma and suggested our cases to be part of the papillary renal cell carcinoma group. Discussion: Papillary architecture is not a feature of renal oncocytoma. In contrast, papillary renal cell carcinomas are defined by their papillary or tubulopapillary architecture. Papillary renal tumors with oncocytic cells might be a distinct variant in the papillary renal cell carcinoma group. All These results suggest that adult papillary renal tumors with oncocytic cells might be a distinct variant in the papillary renal cell carcinoma group. This distinction is of importance because oncocytomas are benign tumors with an indolent course, whereas type 2 papillary carcinomas are malignant tumors associated with a poor outcome. In conclusion, we describe an oncocytic variant of papillary RCC with distinct clinicopathologic features from type 1 and type 2. PP3-100 EXPRESSION OF Ki-67, p53 AND E-CADHERIN IN RAT UROTHELIAL CARCINOGENESIS INDUCED BY NBUTYL-N-(4-HYDROXYBUTYL) NITROSAMINE Paula Oliveira1, Aura Colaco2, Luis De La Cruz3, Carlos Lopes4 1 Department of Veterinary Sciences, CECAV, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal 2 Department of Veterinary Sciences, CECAV, University of Trás-os-Montes and Alto Douro, Vila Real, Portugal 3 Deparment of Physiology, Faculty of Veterinary, Santiago University, Spain 4 Departament of Pathology and Molecular Immunology, Instituto de Ciências Biomédicas de Abel Salazar, University of Porto, Largo Professor Abel Salazar, Porto, Portugal Background: N-butyl-N-(4-hydroxybutyl) nitrosamine (BBN) induced urothelial carcinogenesis is a useful model for studying urothelial tumours. The present study investigated the expression of Ki-67, p53 and E-cadherin in chemically induced rat urothelial lesions. Material and Methods: Female Fisher 344 rats (n=40) were given 0.05% BBN in their drinking water for 10 and 20 weeks and then euthanized. Twenty animals were used as negative control. Results: At the 10th week 100% of rats exposed to BBN exhibited simple hyperplasia, 40% nodular hyperplasia, 90% dysplasia, 20% papiloma, 10% of papillary neoplasm of low-malignant potential, and 20% squamous metaplasia; after 20 weeks 30% of rats had developed simple hyperplasia, 80% nodular hyperplasia, 100% dysplasia, 40% papilloma, 50% papillary neoplasm of low-malignant potential, 70% low-grade carcinoma, 20% high grade carcinoma, 10% invasive carcinoma, and 60% squamous metaplasia. Ki-67, p53 and E-cadherin were analysed immunohistochemically in paraffin embedded tissue. Low- and high-grade papillary carcinoma showed labelling and apoptotic index greater than other lesions (p<0.05). Simple hyperplasia and squamous metaplasia showed a similar Ecadherin pattern when compared with normal urothelium, with their expression confined to the cell membrane. Nodular hyperplasias, dysplasias and papillomas exhibited weak Ecadherin expression. All papillary neoplasm of low-malignant

potential, low-and high-grade carcinoma revealed an increase in cytoplasm reactivity and cell discontinuous membrane positivity. E-cadherin immunoreactivity correlated significantly with p53 index (p<0.05) and Ki-67 (p<0.05) antigen index. Conclusion: We concluded from our data that immunohistochemical Ki-67, p53, E-cadherin staining may provide additional information on rat urothelial carcinogenesis. PP3-101 MALACOPLAKIA OF THE GENITOURINARY TRACT: A CLINICOPATHOLOGICAL STUDY OF 5 CASES Faten Limaiem1, Amina Mekni1, Ines Chelly1, Salma Bellil1, Issam Msakni1, Slim Haouet1, Nidhameddine Kchir1, Mohamed Moncef Zitouna1, Mohamed Horchani2, Khadija Bellil1 1 Department Of Pathology. La Rabta Hospital. Tunisia 2 Department Of Urology La Rabta Hospital Tunisia Introduction: Malacoplakia (MPK) is a rare granulomatous inflammatory disease that predominantly affects the urinary tract (60%) with a predilection for the bladder while renal parenchymal involvement is less common, occurring in 16% of cases. Testicular and prostatic MPK are exceedingly rare. This benign lesion is histologically characterized by histiocytes containing distinct basophilic inclusions called “MichaellisGutmann bodies” which are believed to result from abnormal macrophage function. Patients & Methods: Between January 1999 and February 2007, 5 cases of genitourinary tract MPK involving the prostate (n = 2), the bladder (n = 1), the kidney (n=1) and the testis (n=1) were diagnosed at the pathology department of La Rabta Hospital. Medical records and microscopic slides of each patient were retrospectively reviewed. Results: There were 3 male and 2 female patients aged between 50 and 80 years (mean age = 65,40 years). The chief complaints were fever (n=3), dysuria (n=3), nocturia (n=2), bladder outlet obstructive symptoms (n=2), lumbar pain (n =1) and testicular swelling (n =1). One patient underwent right nephrectomy for a cystic renal mass associated with pyelonephritis. Radical orchidectomy was performed in another patient. A prostatic (n=2) and vesical (n=1) biopsy under ultrasonic guidance was realized in the remainder of cases. Histological examination of the surgical and biopsy specimen showed a granulomatous inflammatory process characterized by the presence within histiocytes (von Hansermann‘s cells) and also extracellularly in the stroma, of concentrically layered, target or ring-like structures that reacted positively to periodic Schiff reagent. The final pathological diagnosis was malacoplakia. During the follow-up period that ranged between 6 months and 3 years, all patients were well and alive. PP3-102 DIAGNOSTIC AND PROGNOSTIC VALUES OF c-KIT, Ki–67, MATRIX METALLOPROTEINASE-1 (MMP-1) EXPRESSIONS IN GERM CELL TUMORS OF TESTIS Sema Kizilgedik Dogdas, O. Fahrettin Goze Cumhuriyet University Faculty of Medicine Pathology Department, Sivas, Turkey Germ cell tumors constitude 90-95% of all the testis tumors. Testicular germ cell tumors may be either in pure or in mixed form. Pure germ cell tumors are divided into two groups as seminomatous (seminoma and spermatocytic seminoma) and non-seminomatous (embryonal carcinoma, yolk sac tumor, trophoblastic tumor and teratoma) tumors. Mixed germ cell tumors include mainly embryonal carcinoma-teratoma, teratomaseminoma, choriocarcinoma and different combinations of the others. H&E stained slides of 49 cases found in the archieves of Cumhuriyet University Faculty of Medicine Pathology Department which were diagnosed as germ cell tumor were reexamined according to the criterias of WHO defined in 2004. Slides obtained from paraffine blocks were stained with PLAP,

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C-kit, Ki-67 and MMP-1 immunohistochemically. When pure and mixed germ cell tumors were reexamined distribution of the tumors were found as 30 cases of seminoma, 25 cases of embryonal carcinoma, 13 cases of yolk sac tumor, 14 cases of teratoma and two cases of spermatocytic seminoma. Statistical analysis of the study was done among the tumors with different histopathological types which also include intratubular germ cell neoplasia. In respect to PLAP and C-kit expression there is a significant difference betweeen seminoma and nonseminomatous tumors which have strong and intermediate membranous staining property (p<0.05). The highest Ki-67 proliferation index was determined in spermatocytic seminoma (0.63±0.01) and embryonal carcinoma (0.50±0.12). The difference between these two tumors and the others having low proliferation index such as seminoma, yolk-sac tumor, and teratoma was also significant (p<0.05). As a result, strong and intermediate membranous staining with PLAP and C-kit is a reliable marker in differential diagnosis between seminoma and non-seminomatous tumors. PLAP expression suggested the diagnosis of IGCN more than C-kit. When we considered the Ki67 staining property, spermatocytic seminoma and embryonal carcinoma had higher proliferation indexes than the other germ cell tumors. Increase in Ki-67 index with an increase in primary tumor stage showed that those markers are important in prognostic aspect. PP3-103 IMMUNOCHEMICAL FRACTAL ANALYSIS OF NEOVASCULAR NETWORK IN PROSTATE CANCER P. Kakouri1, P. Pavlopoulos1, N. Kavantzas2, A. Apostolaki3, E. Koniaris3, S. Bouzoukas4 1 1st Social Security Urological Clinic, Athens, Greece 2 Department of Pathology, National University of athens Medical school, Athens, Greece 3 Department of Pathology, Anticancer Oncological Hospital of Athens 'St. Savvas', Athens, Greece 4 1st Social Security Hospital, Urologic Clinic, Athens, Greece BACKGROUND: Fractal analysis is being applicated in the field of urological pathology with increasing frequency. In the present study, fractal analysis was performed in order to be investigated whether prostate cancer neovascular network fractal dimension (FD) differs from that of normal prostate vasculature. METHOD: Archival material from 34 cases of prostate cancer as well as 10 normal or hyperplastic prostates were examined. From each case two representative CD34 immunohistochemically stained fields (x200) were randomly selected and digitised as high quality JPEG files. Using especially designed image analysis applications the images were automatically processed to threshold the vascular section, by the use of a modified and extended clastering method for colour images based on an algorythm proposed by Otsu. The solid areas of the binary images were then converted to an outline of single pixels, and finally the fractal dimension was estimated using a box - counting algorythm implemented in our application. RESULTS: The normal prostate vasculature presented a mean FD of 1,959. The corresponding value of prostate cancer neovascular network was 1,3297. Both values were statistically different from the topological dimension (= 1,000). The difference of the two values was statistically significant (all p<0,05). CONCLUSION: Our findings suggest that by using the proposed methodology the vascular network of prostate cancer presents fractal charateristics. Thus, FD seems to be an important morphometric parameter possibly valuable for the future development of an unbiased and reproducible image analysis system for the study of prostate cancer pathology.

PP3-104 HYBRID RENAL CELL CARCINOMA WITH EOSINOPHILIC CHROMOPHOBE AND UNDIFFERENTIATED FEATURES DEMONSTRATING A THIRD FEATURE WITHIN IPSILATERAL ADRENAL METASTASES Ozden Tulunay1, Guldal Yilmaz1, Yasar Beduk2 1 Medical School of Ankara University, Departments of Pathology, Ankara, Turkey 2 Medical School of Ankara University, Departments of Urology, Ankara, Turkey Background: Hybrid renal cell carcinoma (RCC), containing tumor cells displaying cytological features of different subtypes in the same kidney has been described recently. Although rare, synchronous renal primary malignancies can be observed. Identification of the hybrid character of an RCC has important diagnostic, and potentially, prognostic ramifications. Case report: A 55-year-old male admitted to the intensive care unit, due to idiopathic myelofibrosis, with left lumbar pain of 20 days duration. His past medical history was unremarkable otherwise, and there was no family history of a hereditery disease. Serological studies demonstrated minimal increase in serum BUN and creatinin levels. Abdominal CT 2 cm solid mass in the midportion of the left kidney and aurevealed a 3.2 1.5×0.9 cm solid mass in the left adrenal gland. Patient underwent a left radical nephrectomy. Findings: The resected left kidney weighed 357 g, with a 10.5×4.5 cm.u2.8×2.7 cm mass in the mid-portion, measuring 17u3.6 Microscopically, the tumor was an eosinophilic variant of chromophobe RCC (ChRCC/eo) with areas of undifferentiated carcinoma (UC). UC with necrosis and high nuclear grade of Furhman (Grade 3), formed multiple foci within ChRCC/eo, and revealed large polygonal cells with condensed eosinophilic/granular cytoplasm with increased Nuclear size, pleomorphism, high mitotic activity, and 4.5×2 cm,udemonstrated transitions with ChRCC/eo. The left adrenal gland, 6.5 2×1.2 cm solid mass, consisted of a tumor displaying cytologicaluwith a 2.7 features of UC and of mucinous tubular carcinoma (MTC). The adrenal vein and periglandular lymphovascular spaces were extensively invaded by UC panCK, CK7, and EMA were diffusely and strongly positive in all tumor subtypes while, CEA E12 only focally, in UC Conclusion: Hybrid RCCsEwas positive only in MTC, and 34 exhibit characteristic histologic features that set them apart from classical RCC. This type of hybrid tumor seems to be associated with a more aggressive behavior, and warrants diagnostic caution. This case is the first of its type reported in the literature, which is a combination of ChRCC/eo, and UC within the same tumor of the kidney and of a synchronous third subtype, MTC, within the synchronous metastases in the ipsilateral adrenal gland. PP3-105 RETROPERITONEAL LYMPH NODE METASTASES FROM BURNT-OUT PRIMARY TESTICULAR SEMINOMA. REPORT OF TWO CASES S. Bellefqih, K. Joseph, A. Durlach, J. Brandt, P. Birembaut Laboratoire Pol Bouin, Departement d’Urologie, CHU Hopital Maison Blanche, Reims-France The burnt-out phenomenon in germ-cell neoplasia is defined by the presence of an extragonadal germ cell tumour with no tumour at the testis. This tumour has spontaneously regressed with no treatment. We report two cases of burnt-out primary testicular tumours revealed by a retroperitoneal metastatic seminoma. Case 1 A 30-year-old man, with a history of cryptorchidy, was admitted for exploration of a left iliac lymph node mass, 7 cm in diameter. A biopsy specimen suggested a metastasis from a germ cell tumour. At clinical examination, the left testis was small. Case 2 A 40-year-old man presented with flank pain. Abdominal CT scan showed a 9 cm mass left retroperitoneal between the aorta and inferior cava. A lymphadenectomy was performed.

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Frozen section analysis suggested lymph node metastasis from testis seminoma. Scrotal ultrasonography in these 2 patients revealed a small left testis containing a 5 mm (case 1) and 1 cm (case 2) hypoechogenic area. Chest CT was normal. They had a left radical orchidectomy and the lymphadenomectomy. The macroscopic examination of the pulp (case 1: 34 g , 3x2 cm; case 2: 46g, 3,5x2,5) showed a 8 mm juxtahilar ( case 1) and 1.5 cm central (case 2) fibrous scar. Histological examination showed a testicular atrophy with sclerosis of the seminiferous tubules, hyperplastic Leydig cells, and foci of intratubular germ cell neoplasia unclassified type. associated with fibrous scar and calcifications. Lymph node (n=3) examination confirmed a pure seminomatous germ cell metastatic tumour with an epithelioid giant cell reaction associated with lymphocyte infiltrate. Immunohistochemical staining in lymph node specimen showed a cytoplasmic membrane immunoreactivity for placental alkaline phosphatase and CD117. Chemotherapy was administred. Testicular ultrasonography is today the gold-standard technique and any parenchymal echostructure abnormality should be verified histologically PP3-106 A SEARCH FOR THE USEFULNESS OF ANTIBODY COCKTAIL (AMACR (P504S)+HMW-CK+P63) IN PROSTATIC NEEDLE BIOPSIES WITH A PROPOSAL OF A NEW SCORING SYSTEM Dilek Demir, Isin Kilicaslan, Yasemin Ozluk, Veli Uysal Istanbul University, Istanbul Faculty of Medicine, Department of Pathology, Istanbul, Turkey BACKGROUND We aimed to search for the usefulness of an antibody cocktail, combining alpha-methylacyl CoA Rasemase (AMACR), high molecular weight cytokeratin (HMW-CK) and p63 antibodies, in differentiating prostate cancer (Pca) from highgrade prostatic intraepithelial neoplasia (HGPIN), atypical small acinar proliferation (ASAP) and benign glands. We also tried to evaluate whether to use HMW-CK singly or to use the antibody cocktail. MATERIALS-METHODS: We included 109 prostatic needle biopsies displaying features of ASAP, HGPIN and Pca in H&E stain. Benign glands were detected in all biopsies as a control group. Each lesion were analyzed separately. HMW-CK and p63, as basal cell markers, were expected to be positive in basal cells of benign glands and HGPIN, whereas AMACR was expected to be positive in the tumoral cells. The result of the interpretation of the antibody cocktail were scored from (-1) to (7) based on the percentage and the intensity of staining. We checked the statistical significance of the diagnostic value of the antibodies, individually and together. RESULTS Thirty five lesions of carcinomas were all negative for basal cell markers, whereas those markers were detected to be positive in HGPIN and benign glands. The staining pattern was patchy or diffuse, respectively. In ASAP lesions, immunohistochemistry demonstrated basal cell staining in 67.7% (23/34) and no staining was found in 32.3% (11/34). It did not differ whether we use p63 or HMW-CK as a basal cell marker. We detected strong positivity for AMACR in carcinomas in more than 75% of the tumor. Strongly positive staining for AMACR in more than 75% of cells was seen in 31.4% (17/54) of HGPIN and 26.4% (9/34) of ASAP. Benign glands were negative for AMACR. The immunoreactivity score of carcinomas were 7 in 32 and 6 in 3, whereas it was •6 in 16.6% (9/54) of HGPIN and 23.5% (8/34) of ASAP. None of the benign tissues displayed a score of •5. We observed a superiority in terms of specificity for antibody cocktail (96.3%) over HMW-CK (85.1%) in ASAP. The sensitivity for both antibodies were 100%. CONCLUSIONS As a result, we suggest that combining AMACR with a basal cell marker may improve diagnostic performance, especially in ASAP lesions. We recommend the use of antibody cocktail as a simple and cost-effective method in routine practice.

Pulmonary Pathology PP3-107 PLEUROPULMONARY ABNORMALITIES IN PATIENTS WITH MULTIPLE MYELOMA Jurgen Knolle1, Stefanie Hege1, Beatrix Bohnsteen2, Jens Schreiber2 1 Dessau Medical Center, Institute for Pathology, Germany 2 Department of Pulmonary Medicine, Dessau, Germany Background: Multiple Myeloma (MM) is a low malignant Non– Hodgkin’s lymphoma. Pleuropulmonary abnormalities in patients with MM require a broad differential diagnostic spectrum. Method: We retrospectively analysed the clinical histories and findings of 22 patients with MM in whom bronchoscopies were performed because of pulmonary infiltrates, both alveolar and interstitial. The series included 16 men and 6 women; the median age was 64 y (46 – 83 y). Results: Bronchoscopy enabled differential diagnosis in all pts. and was performed without complications regardless of cytopenias. In 16 pts. (72,7%) infections were diagnosed. Rare manifestations were diffuse pleural and pulmonary infiltrations with atypical plasma cells (n=1), light chain deposition disease (n=1), diffuse alveolar hemorrhage (DAH) (n=1), amyloidosis (n=1) and toxic pneumonitis (n=2). Subclinical DAH was found in 7 pts. (31.8%). In one patient an intercurrent disease was diagnosed. Conclusion: We conclude that the most common causes of pulmonary abnormalities in pts. with MM are infections. However there are rare differential diagnoses, which are presented in more detail. PP3-108 SOLITARY FIBROUS TUMOUR OF PLEURA- 16-YEARS REVIEW Jelena Stojsic, Branislava Milenkovic, Maja Ercegovac Institute for Lung Diseases and Tuberculosis, Clinical Centre of Serbia, Belgrade, Serbia Background: Solitary fibrous tumour of pleura (SFTP) originate from primitive submesothelial fibroblasts differentiated to hemangipericytoma-, neurofibroma- and fibroma-like pattern. SFTPs are pedunculated with visceral, parietal and mediastinal pleura. Extrathoracic SFTPs are rare. Method: Analysis was made on 47 SFTPs diagnosed on surgery extirpated tumours during last 16 years. Results: SFTPs was diagnosed in 25 females and 22 males. Patients’ age ranged from 27 to 71 years, average 52.1 and predominance in 6. life decade. Eighteen tumours were localized on the right and 13 from the left site and 14 in mediastinum. Twenty tumours originated from visceral, 13 from parietal and 14 from mediastinal pleura. SFTPs Tumour size varied from 30mm to 540mm, mean 117cm. The greatest SFTP measured 4900g, occuping the whole right hemithorax, involving pulmonary tissue, visceral and parietal pleura and ribs. The other giant SFTPs measured 235mm (2430g), 240mm(2850g) and 350 mm (3600g). Predominant morphological pattern according cellularity was: 28 mixed, 11 purely hypo- and 8 purely hypercellular and rarely purely hypo- or hypercellularity. Predominant morphological pattern was: hemangio-19, neurofibro-19 and fibroma-like pattern 9. Three of them were malignant on surgery. Three SFTPs recidived 3 months to 6 years after surgery. Diagnosis of SFTP was precised by histochemical (van Gieson, Masson-trichrome) or by immunohistochemical stains (vimentin, CD34, bcl-2). Preoperative needle biopsy was performed in 17 patients with suggestive diagnosis of mesenchymal tumour of variable morphological pattern and unpredictive biological behavioring. Conclusion: Diagnosis of SFTPs on preoperative biopsy can be estimated by percutaneous needle biopsy applying CD34. Precise diagnosis of SFTPs is useful for planning surgery. Hypercellularity, increased mitotic activity, nuclear polymorphism and invasion of tumour capsule are suggestive for suspicion on malignancy. Tumour size is not

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mandatory for malignancy. On gross examination of tumour node it is necessary to obtain numerous tissue samples specially with tumour capsule for precise the biological behavior of SFTP. PP3-109 PULMONARY ANGIOMYOLIPOMA: CLINICOPATHOLOGICAL ANALYSIS OF 2 NEW CASES AND 10 PREVIOUSLY REPORTED CASES Irem Hicran Ozbudak1, Konstantin Shilo2, Jeffrey R. Galvin3, Teri J. Franks2 1 Department of Pathology, University of Akdeniz School of Medicine, Antalya, Turkey 2 Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology, Washington, DC, USA 3 Department of Radiologic Pathology, Armed Forces Institute of Pathology, Washington, DC, USA; Department of Diagnostic Radiology and Pulmonary/Critical Care Medicine, University of Maryland School of Medicine, Baltimore, MD, USA Background: Angiomyolipoma is a rare mesenchymal tumor that most often arises in the kidney, but can also occur in extrarenal locations. Angiomyolipoma of lung is very rare and to date only 10 cases have been reported in the literature. Method: Three cases of pulmonary angiomyolipoma were identified in the registry of the Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology, from 1977 to 2007 (one of which was previously reported). The clinical, radiologic and pathological findings of these three cases were reviewed and were compiled with findings abstracted from the cases reported in the literature. Results: Twelve patients included 4 males and 8 females (male:female, 1:2) with a mean age of 52 years (range, 28 to 68). Pulmonary angiomyolipoma occurred de novo in 10/12 (83%) patients and in association with tuberous sclerosis in 2/12 (17%) patients. Pulmonary lesions presented as isolated findings in 7/12 (58%) patients, but were also seen in association with renal (3/12 patients, 25%), hepatic (2/12 patients, 17%) and splenic (1/12 patient, 8%) angiomyolipoma. Radiologically, tumors were well-circumscribed nodules with fat density that were located in the lung parenchyma in 10/12 (83%) patients and as an endobronchial lesion in 2/12 (17%) patient. Tumors ranged in size from 0.4 to 9.5 cm (mean, 2.2 cm). Microscopically, all tumors demonstrated variable amounts of mature adipose tissue, smooth-muscle cells and tortuous, small- to medium-sized, thickwalled blood vessels. Minor histologic features included: epithelioid smooth-muscle cells containing melanin pigment, histiocytes, multinucleated giant cells, plexiform vascular channels, atypia and mitosis. Tumor smooth-muscle cells were immunoreactive with HMB-45 (6/9, 67%), actin (6/7, 86%), desmin (2/4, 50%) and S-100 (2/6, 33%). Conclusion: Pulmonary angiomyolipomas are rare tumors with only 12 cases known to date. They show a female gender bias. Pulmonary angiomyolipomas typically arise de novo as parenchymal nodules. The combination of thick-walled vessels, smoothmuscle cells and adipose tissue is distinctive and aids separation from other benign and malignant pulmonary mesenchymal lesions. PP3-110 EXTRAMEDULLARY HEMATOPOIESIS IN PULMONARY SPINDLE CELL TUMORS: RARE AND UNUSUAL ASSOCIATION Irem Hicran Ozbudak1, Konstantin Shilo2, Jeffrey R. Galvin3, Teri J. Franks2 1 Department of Pathology, University of Akdeniz School of Medicine, Antalya, Turkey 2 Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology, Washington, DC, USA 3 Department of Soft Tissue Pathology, Armed Forces Institute of Pathology, Washington, DC, USA

Background: Extramedullary hematopoiesis is defined as development and growth of hematopoietic tissue outside the bone marrow. It tends to occur as a compensatory phenomenon in the organs of reticuloendothelial system. Involvement of other organs and some tumors such as pilomatricoma and hemangioblastoma is recognized. Only one case of pulmonary carcinoid with intratumoral hematopoiesis is reported in the English language literature. Method: Two cases of lung tumors with extramedullary hematopoiesis were identified in the registry of the Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology. Herein, we describe the pathological and immunohistochemical features of these two cases. Results: A 72 year-old male presented with 3.5 cm right middle lobe, pleural based solitary fibrous tumor. The tumor was composed of CD34 positive spindle cells with variable collagen deposition. Scattered nests of extramedullary hematopoiesis were distributed throughout the lesion. They exhibited trilineage hematopoiesis and contained numerous dysplastic megakaryocytes. A 64 yearold male presented with a 7 cm peripheral left lower lobe intermediate grade sarcoma with myxoid features. Tumor cells were immunoreactive for smooth muscle actin, muscle specific actin and vimentin, while the pancytokeratin, CAM5.2, EMA, desmin, HMB-45, S-100, CD34, Factor VIII were negative. Hematopoietic cells consisting of megakaryocytes, erythroblasts and rare myeloid precursors were present as small clusters throughout the tumor. No hematopoietic cells were identified in vessels or adjacent lung parenchyma in either case. Megakaryocytes and erythroid precursors were immunoreactive with Factor VIII and hemoglobin respectively. Conclusion: Tumors of the lung displaying foci of intratumoral extramedullary hematopoiesis are rare. However, awareness of this association is helpful in avoiding misdiagnosis of a hematopoietic malignancy and upgrading of spindle cell tumors based on the dysplastic megakaryocytes which resemble pleomorphic tumor cells. PP3-111 INFLAMMATORY MYOFIBROBLASTIC TUMORS OF THE LUNG ARE NEGATIVE FOR HHV-8 Fabio Tavora1, Irem Hicran Ozbudak2, Konstantin Shilo1, Jean M. Przybocki3, Guanghua Wang3, William D. Travis4, Teri J. Franks1 1 Department of Pulmonary and Mediastinal Pathology, Armed Forces Institute of Pathology, Washington, DC, USA 2 Department of Pathology, University of Akdeniz School of Medicine, Antalya, Turkey 3 Department of Molecular Pathology, Armed Forces Institute of Pathology, Washington, DC, USA 4 Department of Pathology, Memorial Sloan Kettering Cancer Center, New York, NY, USA Background: Inflammatory myofibroblastic tumors are uncommon lesions composed of spindled myofibroblasts within a variable fibroinflammatory background. Although the true nature of these lesions has not been fully elucidated, identification of consistent cytogenetic alterations in the anaplastic lymphoma kinase (ALK) gene suggests that they may represent a neoplastic proliferation. Furthermore, few cases of inflammatory myofibroblastic tumor have been reported to harbor human herpesvirus (HHV)-8 infection, implicating HHV-8 in its pathogenesis. In this study, the largest to date series of pulmonary inflammatory myofibroblastic tumors were tested for the presence of HHV-8 by immunohistochemical and molecular methods. Method: Twenty cases with classical features of inflammatory myofibroblastic tumor were selected for the study. Immunohistochemical studies for latent nuclear antigen of HHV8 (clone 13B10, 1:50, Vector Laboratories, Burlingame, CA) and for ALK (clone ALK-1, 1:100, Dako, Carpinteria, CA) were performed according to the manufacturers’ instructions. Four open reading frames (ORF) of HHV-8, including ORFK2, ORF16, ORF26, ORF72, were targeted with real time PCR.

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Results: The study cohort included 9 men and 11 women with a mean age of 37 years (range, 1 to 81). Microscopically, the tumors were composed of predominantly spindled myofibroblasts. On immunohistochemical studies, 4/20 cases (20%) demonstrated diffuse cytoplasmic positivity with ALK-1. Imunohistochemical staining for HHV-8 was negative (0/20, 0%) in all cases. All inflammatory myofibroblastic tumor cases tested with real time PCR were negative (0/20, 0%) for all four ORF, while 10/10 (100%) samples of Kaposi sarcoma, serving as positive control, were positive. Her2 gene, tested for the presence of amplifiable DNA in the tissue lysate, was positive in all (20/20, 100%) of the inflammatory myofibroblastic tumors. Conclusion: This study documents the absence of HHV-8 in a series of pulmonary inflammatory myofibroblastic tumors and suggests that further investigation is needed in elucidating other potential etiologic factors for this lesion. PP3-112 IDENTIFICATION OF RELEVANT LUNG ADENOCARCINOMA BIOMARKERS BY MASS SPECTROMETRY BASED N-GLYCOPROTEIN PROFILING OF PLEURAL EFFUSIONS Alex Soltermann1, Reto Ossola2, Sandra Kilgus-Hawelski1, Arnold Von Eckardstein3, Tobias Suter4, Ruedi Aebersold2, Holger Moch1 1 Institute for Surgical Pathology, University Hospital Zurich, CH-8091, Switzerland 2 Institute for Molecular Systemsbiology, Swiss Federal Institute of Technology, CH-8093, Switzerland 3 Institute for Clinical Chemistry, University Hospital Zurich, CH-8091, Switzerland 4 Clinics of Immunology, University Hospital Zurich, CH-8091, Switzerland Background: Late stage lung adenocarcinoma is frequently accompanied by pleural effusion. Tumour cells in the effusion may be the source of proteins, which are potentially detectable in tumour tissue or serum. In particular, N-glycosylated proteins (NGP) can be secreted into body fluids and are therefore considered potential biomarkers. We aimed for establishing a mass spectrometry based N-GP effusion profile for cytology specimens. Methods: Malignant pleural effusions of 5 patients with lung adenocarcinoma and 5 non-malignant controls were used for N-GP capturing by solid phase extraction. Following trypsin digest and PNGase F release, a liquid chromatography separation connected online to a tandem mass spectrometer was performed. The resulting peptide spectra were screened against IPI and NCI databases. Results: Concentration differences between malignant and non-malignant effusions were found for total protein, LDH, cholesterol, glucose, CA-125 and CEA (p’s < 0.05). 174 non-redundant N-GP were detected in the total of 10 samples with probability • 0.9, 29 of them exclusively in malignant effusions. The overall specificity for the N-glycomotif was 89%. Identifications were compared with serum of 4 healthy donors and A549 cell culture supernatant as well as published data sets. Specific protein markers were correlated with their corresponding immunoreactivity in either effusion fluid (CA-125, interferon gamma) or tumour cells or tissue (periostin, CD166), respectively. By applying strict criteria and using data mining, a potential lung adenocarcinoma marker signature of 6 proteins, including periostin, CD166, multimerin 2, lysosome-associated membrane glycoprotein 2, latent TGF beta binding protein 2 and surfactant protein A, was elaborated. Conclusion: The N-GP capturing and LC-MS/MS protocol enables the confident identification of several relevant biomarkers for lung adenocarcinoma.

PP3-113 VALUE OF VARIOUS CYTOHISTOLOGICAL TECHNIQUES FOLLOWING FIBEROPTIC BRONCHOSCOPY IN THE DIAGNOSIS OF LUNG CANCER Nicoletta Maounis1, Eleni Ellina1, Aikaterini Blana1, Maria Chorti2, Agathi Lekakou3, Nikos Trakas4, Aikaterini Pierakou2, Aphroditi Emmanouilidou1 1 Department of Clinical Cytopathology. General Hospital,Greece 2 Department of Pathology General Hospital, Greece 3 1st Pulmonary Department General Hospital, Greece 4 Department of Biochemistry General Hospital, Greece Introduction: The aim of the present study was to evaluate the value of various diagnostic techniques following fiberoptic bronchoscopy in the diagnosis of lung carcinoma. Material and methods: A total of 2005 patients underwent diagnostic bronchoscopic procedures during a 3-year period for various neoplastic and nonneoplastic lung diseases. Histologic examination of specimens obtained by forceps biopsy and cytological evaluation on brushing, washing and postbronchoscopic sputum smears were performed. The diagnostic value of the procedures was analyzed by Mc Nemar’s exact test. Results: Eight hundred thirty patients had at least one of the techniques positive for lung malignancy (41,39%). Mean age 67.46; range 22-91 years. %). Of the above 830 tumors, 302 were diagnosed as squamous cell carcinoma (36.4%), 134 as adenocarcinoma (16.1%), 11 as BAC (1.3%), 148 as small cell lung carcinoma (17.8%), 191 as non- small cell lung carcinoma (23%), 8 as mixed (1.0%),12 as metastatic (1.4%), 4 as carcinoids (0.5%) 1 as lymphoma (0.1%) and 19 as suspicious for malignancy (2.3%). Forceps biopsy specimens gave positive results in 34.3% of our malignant cases, brushing in 52.8%, washing in 69.4% and post-bronchoscopic sputum in 28%. Collection of brushing specimens in addition to washing and post-bronchoscopic sputum increased the diagnostic yield of bronchoscopy from 78.8% to 87.5% (p=0.0001). Furthermore, the addition of forceps biopsy increased the diagnostic yield of the combination of the three cytological techniques from 87.5 to 98.8% (p<0.0001). In conclusion, the best diagnostic yield for the diagnosis of lung cancer, in our material which included visible and non-visible pulmonary lesions, was obtained with the association of brushing and bronchial washing specimens. Furthermore, the application of both cytology and histology can achieve better results. PP3-114 PROGNOSTIC SIGNIFICANCE OF VASCULAR AND INTERSTITIAL REMODELING PROCESSES IN NONSPECIFIC INTERSTITIAL PNEUMONIA: SYSTEMIC SCLEROSIS VERSUS IDIOPATHIC Erika De Carvalho, Edwin Parra, Romy De Souza, Alexandre A´B Saber, Juliana Machado, Vera Capelozzi University of São Paulo Medical School, Departments of Pathology and Rheumatology, Brazil Aims and Methods: Pulmonary biopsy specimens were examined from 40 patients, 22 with idiopathic nonspecific interstitial pneumonia (NSIP) and 18 with nonspecific interstitial pneumonia associated with systemic sclerosis (SSc). We compared the septal and vascular matrix remodeling, vascular grade, pulmonary function tests and survival between the two groups. Results: The content of septal collagen and elastic fibers, as well as the elastic fibers in the vascular interstitium, were higher in the SSc group ( p=0.01, p=0.001 and p<0.0001, respectively). Among pulmonary function tests the DLCO/VA was affected to a greater extent in the SSc group (59% of the predicted value in SSc and 97% in the idiopatic group). There were no differences in the collagen content of the vascular interstitium, vascular grade, or survival between the two groups. Conclusions: Although the fibrotic process is more intense in the SSc group, it does not affect the

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prognosis of these patients, as it has been described in idiopatic lung fibrosis. It seems that the fibrogenic process has a minor role in SSc when compared with the idiopatic group. Because the elastotic process is higher in the SSc group, this might suggest that autoimmune inflammatory mechanisms affecting the elastic fiber system could play a greater role in the pathogenesis and pulmonary remodeling process of SSc-NSIP than in idiopathicNSIP. PP3-115 PARENCHYMAL AND VASCULAR INTERACTIONS IN THE PATHOGENESIS OF NON SPECIFIC INTERSTITIAL PNEUMONIA IN SYSTEMIC SCLEROSIS AND IDIOPATHIC INTERSTITIAL PNEUMONIA Edwin Parra, Erika De Carvalho, Romy De Souza, Alexandre A´B Saber, Vera Capelozzi University of São Paulo Medical School,Departments of Pathology and Rheumatology, Brazil Background: Interstitial lung disease is a well recognized prognostic factor in systemic sclerosis (SSc). As the prognosis in nonspecific interstitial pneumonia (NSIP) has been described to be better in collagen vascular disorders compared to the idiopathic forms, we hypothesize that the mechanisms of repair and remodeling are different between these two forms of the disease. Objectives: To compare the mechanisms of repair and remodeling between SSc associated nonspecific pneumonia and the idiopathic form, its impact on pulmonary function tests and survival rates. Methods: We analyzed 18 biopsies from patients with NSIP associated with SSc and 22 with idiopathic NSIP and compared the epithelial and vascular densities as well as vascular activity by immunohistochemistry with antibodies directed against Cytokeratin-7, Surfactant protein-a, CD34, and VCAM-1. Results: Epithelial cell density was lower in SSc-NSIP when compared with idiopathic-NSIP (p<0.0001). Type II pneumocytes and Clara cells were reduced in idiopathic NSIP ( p=0.02) . A decrease in microvessel density was found in SScNSIP compared to idiopathic-NSIP (p<0.0001). The vascular activity measured by VCAM expression was higher in NSIP-SSc when compared to the idiopathic group (p<0.0001). The DLCO/VA in SSc-NSIP was more compromised . A direct association between vascular density and DLCO/VA was found (p=0.02). There was no difference in the survival rate between the two groups after a follow-up of 36 months. Conclusions: Alterations in the epithelium and vasculature seem to differ in the pathogenesis of SSc-NSIP when compared to the idiopathic form of the disease. Further studies may be required to assess the significance of these findings and explore if they can provide prognostic and/or treatment information. Financial Support: FAPESP,CNPq. PP3-116 AIRWAY COLLAGEN AND ELASTIC FIBERS REMODELING IN IDIOPATHIC INTERSTITIAL PNEUMONIAS Edwin Parra, Gustavo Noleto, Vera Capelozzi, Marco Antonio Capelozzi University of São Paulo Medical School, Department of Pathology, Brazil Background: Structural alteration of the airways and lung parenchyma, "remodeling", is a recognized feature of pulmonary fibrosis. In this study, we sought to validate the importance of bronchiolar remodelling and to study the relationships between bronchiolar collagen/elastic system and survival in the major histological pattern of idiopathic interstitial pneumonias (IIPs). Material and Methods: We examined bronchiolar collagen/elastic system fibres in 9 non-specific interstitial pneumonia (NSIP), 24 idiopathic pulmonary fibrosis (IPF), 5 bronchiolites obliterants (OP) and 18 acute interstitial pneumonia (AIP) cases. We used the Picrosirius-polarization method and Weigert's resorcin-

fuchsin histochemistry to evaluate the amount of bronchiolar collagen/elastic system fibres. Results: The bronchiolar measurement of collagen fibres content was higher in AIP (p=0.08) than lungs of OP, UIP and NSIP. In addition, was a not difference of elastic fibres inter groups. Conclusion: We concluded that a not differences of progressive bronchiolar fibroelastosis occurs in IIP histological patterns, probably indicating who not interview in to parenchymal injury.Financial Support: FAPESP,CNPq. PP3-117 TYPE V COLLAGEN PARTICIPATION IN PATIENTS WITH SYSTEMIC SCLEROSIS: PRELIMINARY STUDY Edwin Parra, Walcy Teodoro, Vera Capelozzi University of São Paulo Medical School, 1Department of Pathology and Discipline of Reumathology, Brazil Background: Systemic sclerosis (SSc) is a polymorphic and heterogenic systemic disorder with inflammation, fibrosis and vascular damage. Recently, the pathogenesis of SSc has been extensively studied regarding its autoimmunity aspects related to extracellular matrix remodelling, with an emphasis on the collagens at the inflammatory site. The present paper describes the importance of type V collagen morphologic disorganization, distribution and quantitation in pulmonary interstitium in patients with SSc. Methods: We used immunofluorescence and morphology analise to evaluate and quantified the deposition of collagen I, III and V, and correlation with the active alveolar septal, bronchiolar and arterial interstitum remodeling in open lung biopsies of 11 SSc. Results: The interstitum of alveolar septal, bronchiolar and vascular changes were characterized by morphologic disorganization of fibrillar collagen with diverse disarray, thickness and strong green birefringence of the type V collagen. The quantification of alveolar septal, bronchiolar and vascular collagen I was higher in all groups. In addition, the increase of collagen V in vascular interstitum was major than to collagen III (p=0.05), in contras the collagen III septal and bronchiolar interstitum was major than to collagen V. Conclusions: We concluded and suggested that the collagen type V fibrils are increased, in disarray and combine to other collagen (I and III) make larger than normal lungs, creating an abnormal spatial organization principally in vascular interstitium. Financial Support: FAPESP. PP3-118 STATISTIC ANALYSIS OF MORTALITY AND DISEASE INCIDENCE IN CHRONIC LUNG DISEASES AMONG THE POPULATION OF SEMIPALATINSK REGION WITH LONG-TERM RADIOACTIVE AND POLLUTED ENVIRONMENT Gulmira Sagindikova, Evguenija Kogan Moscow Medical Academy, Russia Work is aimed at examining the prevalence of chronic pulmonary pathology, characteristic of mortality and disease incidence in chronic lung disease (CLD) among the population of Semipalatinsk region having been subjected to long-term radioactive and polluted environment. Public health authorities 1969-2003 reports on spread, mortality and disease incidence of respiratory apparatus among the population of the Semipalatinsk region have been analyzed. 7274 habitants of the Semipalatinsk region have been screened and questionnaired and frequently revealed CLD types in the Semipalatinsk region have been epidemiologically researched. 631 autopsy notes with CLD diagnosis of Morbid Anatomy Bureau of Semipalatinsk for 19962003 years period has been studied. Research has revealed tendency to increase in disease incidence and CLD mortality in the course of growth of exposure rate and close location to testing area. Conclusion: tendency to increase in CLD incidence within 1992-2003 years period, i.e. after nuclear tests cessation has been revealed that proves the radiation value as etiopathogenetic factor.

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PP3-119 SUDDEN DEATH CASES DUE TO TUBERCULOSIS Alexandra Enache1, Fotios Chatzinikolaou2, Natalia Vladika2, Ioannis Moisidis2 1 Department of Forensic Medicine,University of Timisoara, Romania 2 Department of Pathology,Theageneio,Cancer Hospital,Thessaloniki,Greece Aim:in this study we investigate the sudden death cases caused by tuberculosis with different localisations. Materials and methods: We performed a retrospective analysis on the forensic necropsies from IML Timisoara in a period of four years 2002 2005. There were 36 cases of death by pulmonary tuberculosis (1.23% of the cases). Results: the majority of the victims, 94.44% were males. 27.77% of the victims were aged between 40 and 49 years. Homeless victims’ cases represented 27.77% of the cases, 73.33% of these not being hospitalized for this disease. On 18 of the cases (50%) we found fibrocavitary form, 12 of the cases (33.33%) has fibronodular form, 4 cases (11.11%) has an ulcerative form and 2 case was of miliar form (5.55%). Some cases presented unassociated pulmonary tuberculosis, some presented pulmonary tuberculosis associated with bronchopneumonia (12 cases), 1 case presented tuberculosis associated with pulmonary carcinoma, some presented pulmonary tuberculosis associated with tuberculosis pericardic and hepatic granular tuberculosis (1 single case) and an association of pulmonary tuberculosis and hepatic tuberculosis granulia. In 61.11% of the cases (22 cases) the pulmonary tuberculosis was associated with hepatic steatosis or cirrhosis and 4 cases (45%) had increased alcohol rate. We did not observe any evidence suggesting the existence of HIV. Conclusions: The study shows that sudden death caused by pulmonary tuberculosis mainly affects male alcohol consumers and social cases. It is necessary to observe. the possible cohabitation of HIV/bK. PP3-120 A MIXED SQUAMOUS CELL AND GLANDULAR BRONCHIAL PAPILLOMA: A CASE REPORT Ferda Aksoy1, Gokhan Haciibrahimoglu2, Irfan Yalcinkaya2, Murat Keles2, Leyla Yagci Tuncer3 1 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital, Department of Pathology Istanbul, Turkey 2 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital, Department of Thoracic Surgery, Turkey 3 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital, FDepartment of Pulmonology, Turkey Bronchial papillomas are very rare representing less than 0.5 % of lung tumours. The histological types are squamous, glandular and mixed. Papillomas should be differentiated from malignancies by endobronchial biopsy, because complete resection appears to be curative. We describe a case of bronchial mixed squamous cell and glandular papilloma of the bronchus diagnosed by frozen section. The case is a 51 years old male smoker, who was admitted with dyspnea and hemoptysis. Physical examination revealed no abnormalities. Thorax computed tomography slices showed a minimal consolidated lesion in left lower lobe. In bronchoscopic examination a swinging polypoid tumour in left main lower bronchus was detected. Bronchoscopic biopsy was not diagnostic. A left thoracotomy was performed for diagnosis and treatment. The lesion which was 1x1.5x 2 cm in size was removed and a frozen section examination was performed. The lesion was diagnosed as bronchial papilloma with frozen section. Postoperative histopathological examination confirmed the diagnosis as “mixed squamous cell and glandular papilloma”. This extremely rare encounter of mixed squamous cell and glandular bronchial papilloma is presented with the review of the literature.

PP3-121 PULMONARY MALT LYMPHOMA: 3 CASE REPORTS Ferda Aksoy1, Erdal Okur2, Asim Kutlu2, Haluk Calisi3, Ebru Sulu3, Semih Halezeroglu2 1 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital, Department of Pathology Istanbul, Turkey 2 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital Department of Thoracic Surgery, Turkey 3 Sureyyapasa Chest Diseases and Thoracic Surgery Training and Investigation Hospital Department of Pulmonology, Turkey Primary pulmonary lymphomas are relatively rare tumours that comprise less than 1 % of all lymphomas. Approximately 70-80 % of primary pulmonary lymphomas are marginal zone B-cell lymphomas of Mucosa-Associated Lymphoid Tissue (MALT) type.In this paper 3 MALT lymphoma cases are presented. Case I: A 52 years old male patient was admitted with a solitary pulmonary nodule of 2 cm in diameter. Transthorasic needle aspiration was performed. Cytologic examination revealed atypical lymphocytes which reminded a low grade lymphoma. A thoracotomy was performed and the lesion was removed with wedge resection. Case II: A 56 years old male patient was referred with solitary pulmonary nodule of 2.5 cm. in diameter. The lesion was removed with thoracotomy and frozen section was performed which revealed tumour of lymphoid tissue. Histopathological examination of both cases revealed marginal Zone B-cell lymphoma of the Malt type. Case III: A 37 years old female was admitted with cough and dyspnea. CT examination showed multiple bilateral patchy consolidations. Transbronchial biopsy was negative. VATS biopsy was performed. Histopathological examination revealed marginal zone B-cell lymphoma of the MALT type. All cases were referred to medical oncology clinic. Pulmonary lymphomas could be found as solitary or multiple nodules in radiographic examination, so it should be reminded in diagnosis of solitary pulmonary nodule and interstitial lung disease. PP3-122 DIFFERENT HISTOLOGICAL PATTERN IN SYSTEMIC SCLEROSIS WITH INTERSTITIAL LUNG DISEASE (ILD) Romy Souza1, Edwin Parra2, Claudia Borges1, Vera Capelozzi2, Jorge Kavakama3, Ronaldo Kairalla4, Eloisa Bonfá1 1 University of São Paulo Medical School, Department of Rheumatology, Brazil 2 University of São Paulo Medical School, Department of Pathology, Brazil 3 University of São Paulo Medical School, Department of Radiology, Brazil 4 University of São Paulo Medical School, Department of Pneumology, Brazil Background: The new histological pattern of idiopathic interstitial pneumonia (IIP) named Centrilobular Fibrosis (CLF) characterized by an aggressive bronchocentric scarring, basophilic intraluminal contents, and consequent parenchyma involvement may be relevant in systemic sclerosis (SSc). In fact, the high frequency of esophageal involvement in SSc patients is compatible with the suggested mechanism of CLF proposed by the authors, which is the pulmonary aspiration caused by the gastroesophageal reflux (GER). Materials and Methods: Twentyeight consecutive patients who fulfilled the ACR criteria for SSc and had ILD on HRCT had been recruited to an open-lung biopsy as part of an ongoing prospective treatment protocol. Sections had been analyzed by two pathologists specialized in lung diseases and classified according to the new consensus classification of IIP and to the diagnostic criteria for CLF. Results: NSIP and CLF pattern had been the predominant patterns in SSc patients, observed in 67.8% and 21.4%, respectively. Intraluminal basophilic content has distinguished the two main groups. In fact, all 6 patients with CLF had this

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feature whereas in only 6 of the 19 NSIP patients this parameter had been observed (p=0.007). Higher frequency of foreign bodies and bronchocentric distribution of the lesions in patients with CLF had been observed when compared to those with NSIP, but not reach statistical significance (p>0.05). As expected, peripheral injury had been found absent in 100% of CLF cases, compared with a frequency of 57.9% in the NSIP, though it had not reaching statistical significance (p>0.05). Other three characteristics had been found in the totality of the CLF patients: epithelium necrosis, basement membrane exposure and bronchiectasia. Conclusion: This is the first report of CFL in ILD of SSc patients, characterized by an intraluminal basophilic content, high frequency of foreign bodies and a bronchocentric distribution of the lesions. Since chronic aspiration had been suggested as the main pathophysiological mechanism for this severe form of lung involvement, the identification of this subgroup of patients will certainly contribute for a more appropriate therapeutic approach. Financial Support: FAPESP. PP3-123 CILIARY DYSMORPHOLOGY: ULTRASTRUCTURAL FINDINGS IN 12 PATIENTS Gulay Ozbilim1, Aysen Uguz2, Irem Hicran Ozbudak1, Fatih Celmeli2, Hakan Er 3 1 Akdeniz University, School of Medicine, Department of Pathology, Antalya, Turkey 2 Akdeniz University, School of Medicine, Department of Pediatrics, Antalya, Turkey 3 Akdeniz University, School of Medicine, Department of Histology, Antalya, Turkey Background: Various ciliary abnormalities are found in patients with nonneoplastic diseases,such as unexplained recurrent upper and /or lower respiratory tract infections in children with or without Kartegener ‘s triad (sinusitis,bronchiectasis and situs inversus). Primary ciliary dysmorphology is an autosomal resessive disorder in childhood. Abnormal cilia which have specific electron microscopic features can be detected by transmission electron microscopic study. Method: Herein, we investigated 37 children with severe chronic /recurrent upper and lower respiratory tract problems for presence of primary ciliary dysmorphology. A plastic nasal brush was used to collect the tissue samples from nasal mucosa. Nasal brushing materials were evaluated by transmission electron microscopic. Other possible etiologic causes of severe and recurrent upper and lower respiratory tract problems were also evaluated by immunological tests, including atopy,respiratory function test, sweat test,reflux sintigraphy and radiologic studies. RESULT: Primary ciliary dysmorphology was shown in 12 of 37 children electronmicroscopically. Mean age of the patients was 11 years at diagnosis. Five of them were male and seven of them were female. Family history of cousin marriages was determined in 4 patients. Six patients had bronchiectasis ,6 patients had chronic sinusitis, 1 patient had gastroeosophageal reflux, 2 patients had situs inversus (including Kartegener’s triad). Ultrastructural examination of samples from nasal brushing revealed at least one of the following abnormalities: 1)Loss of normal 9+2 microtubules pattern , 2)Compound cilia, 3)Multiple axonemes in a single cilium, 4)Dysorganization of axonemes including loss of dynein arms, 5)Increasing in number of intracellular cilia. Conclusion: Primary ciliary dysmorphology is a cause of recurrent upper and /or lower respiratory tract infections in approximately one third of the patients. Electronmicroscopic study of nasal brushing is the valuable method in diagnosis of PCD patients with or without Kartegener’s triad and it may be recommended for unexplained recurrent upper and /or lower respiratory tract infections

PP3-124 EXPRESSION OF THE c-KIT AND Ki–67 PROLIFERATION INDEX IN PATIENTS WITH LUNG CANCER Cumhur Ibrahim Basorgun1, Gulay Ozbilim1, Irem Hicran Ozbudak1, Alpay Sarper2, Abdullah Erdogan2 1 Department of Pathology, School of Medicine, Akdeniz University, Antalya, Turkey 2 Department of Thoracic Surgery, School of Medicine, Akdeniz University, Antalya, Turkey Background: Kit is a transmembran tyrosine kinase receptor which is the product of protooncogene c-kit. Stem cell factor, ligand of c-kit, is co-expressed in various solid tumors, including carcinomas of the lung. The aim of this study was to determine the expression of c-kit and Ki-67 labelling index and the differences between histopathologic subtypes. Methods: In this study, resection materials of 75 lung carcinomas were used. Sections were taken from parafin-embedded blocks and stained immunhistochemically with c-kit and Ki-67 antibodies. Cytoplasmic and membranous staining density and percentage of stained areas were evaluated semiquantitatively. Mann-Whitney U and chi-square test were used for statistical analyses. Clinical data, information on tumor stage and histomorphologic details were reviewed. Results: Seventy-five patients included 70 males and 5 females (male:female, 14:1). Tumor samples of 30 squamous cell carcinomas, 30 adenocarcinomas and 15 small cell carcinomas were studied. Membranous or cytoplasmic c-kit immunoreactivity was documented in 9 of 15 small cell carcinomas (60%), 26 of 30 squamous cell carcinomas (86%) and 25 of 30 adenocarcinomas (83%). The average of Ki-67 labelling index was found 82% in small cell carcinomas, 62% in squamous cell carcinomas, 34% in adenocarcinomas and statistically significant differences were shown between histologic subtypes (p=0.000). No significant association was found between c-kit immunoreactivity and Ki-67 labelling index (p=0.706). Conclusions: C-kit is expressed in a high percentage of lung tumors; however there is no association between c-kit expression and histologic tumor subtypes. Ki -67 labelling index is higher in small cell carcinomas. Among non small cell lung carcinomas, Ki-67 labelling index is higher in squamous cell carcinomas than adenocarcinomas. There is no correlation between c-kit expression and Ki-67 labelling index in lung tumors. PP3-125 ROENTGENOMORFOLOGIC PREDICTORS OF HORMONAL THERAPY RESPONSE IN PULMONARY SARCOIDOSIS Oksana Kichigina, Evguenija Kogan, Vera Osipenko Moscow Setchenov Medical Academy, Russia Sarcoidosis is systemic, relatively high-quality granulomatosis of unknown etiology, which is characterized by the accumulation of T-lymphocytes and mononuclear phagocytes, by the formation of the non-secreting, non-caseous epitelioid-cell granulomas and by the disturbance of the normal architecture of the diseased organ. There is a large quantity of sarcoidosis classification, which do not always accurately reflect the nature of clinical and roentgenological data with the answers to the ongoing therapy. Aim. The aim of the study was to investigate correlation of the Xray and morphological variants of lung sarcoidosis depending on the nature of granulomatous inflammation and special features of answers to the anti-inflammatory hormonal therapy. Materials and methods. High resolution multispiral CT scan and open lung biopsies of lung and lymph nodes have been studied from 40 patients with sarcoidosis. Results. Patients were divided into two groups according to the localization of sarcoid granulomas. First group (27 patients) had perivascular and subpleural arrangement of granulomas – places of pulmonary lymphatic collectors, the second group (13) – did not have subpleural and perivascular granulomas , granulomas were localized predominantly in

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interstitium of respiratory part of the lung. Positive response to therapy has been found in 22 patients of the first group(80%) and 5 patients (20%) didn’t show negative dynamics. In 2 group positive dynamics was observed only in 4 patients (34%) and 9 (66%) didn’t show any. Conclusions. Hormonal therapy response in patients with sarcoidosis correlates with granuloma location and is better when sarcoid granulomas are accumulated in lymphatic collectors of the lung. PP3-126 GEOGRAPHY OF MESOTHELIOMA AND ASBESTOS EXPOSURE Claudio Bianchi, Tommaso Bianchi Center for the Study of Environmental Cancer - Italian League against Cancer - Monfalcone, Italy BACKGROUND The etiological relationship between exposure to asbestos and malignant mesothelioma is well established. However, it remains uncertain if the effects of asbestos are the same in all the populations. We collected the available data about the geographical distribution of mesothelioma. METHODS Some 300 researchers (including epidemiologists, pathologists, clinicians, etc.) were interviewed by a questionnaire to obtain information on mesothelioma incidence, and on the percentage of asbestos-related mesotheliomas in the various countries. In addition, literature data on mesothelioma were reviewed. RESULTS 1) Reliable data on mesothelioma incidence/mortality are available for a relatively low number of countries, corresponding to about 15% of the world population. 2) The highest annual crude incidence rates (over 25 cases per million) are reported or estimated for few countries (Australia, Belgium, UK). Intermediate rates (11-25 cases per million) are estimated for some countries of Europe (France, Germany, Italy, Scandinavian countries, The Netherlands), and New Zealand. Crude incidence rates lower than 11 cases per million are estimated for many countries of Europe (central Europe, Iberian peninsula, Ireland), some countries of America (Canada, USA), Asia (Cyprus, Israel, Japan, Turkey), and Africa (Morocco, Tunisia). 3) A parallelism between mesothelioma incidence and asbestos consumption in previous decades is observed in a large part of countries. 4) Marked differences in mesothelioma incidence from one region to another are seen in several countries (Croatia, Italy, Japan, Spain, Sweden, UK, etc.), with case clustering in areas with shipyards and asbestos-cement factories. 5) Very low incidences of mesothelioma are reported from some countries with high asbestos production and/or consumption, such as Russia, Thailand, South Korea. 6) Major difficulties are encountered in the histological diagnosis of mesothelioma. In some countries the original diagnosis of mesothelioma has not been confirmed by expert panels in a high proportion of cases. In addition, major difficulties are often encountered in mesothelioma registration. CONCLUSIONS Differences in previous asbestos exposure may explain a large part of the differences in mesothelioma geography. The apparent discrepancies between asbestos consumption and mesothelioma incidence have probably to be attributed to different factors (underestimation, low life expectancy, recent history of industrialization), rather than to ethnic differences in susceptibility to asbestos. PP3-127 CONGENITAL MALFORMATIONS OF THE LUNGS AS POSSIBLE PRECURSOR OF CANCER Ludmila Shestakova1, Evgenia Cogan2, Michail Paltsev3, Yurij Cotlovsky4 1 Moskow Setshenov Medical Academy, Russia 2 Moscow Setshenov Academy, Russia 3 Moscow Setshenov Medical Academy, Russia 4 Krasnojarsky Medical Academy, Russia

Aim. A retrospective study was carried out on 110 cases with malignant lung tumors arising within congenital malformation areas. Matherial. There were 91 males / 82 % / and 19 females / 18 % / with ages ranging from 23 to 65 years. In 18 cases / 22% / congenital lung malformations had been revealed 1-15 years earlier lung cancer development. Methods included routine histology, immunohistochemistry of oncomarkers ( p53, bcl-2, cfos, c-myc, K-ras, c-erbB2, TTF-1, CK 8,19, AE1/AE3, Ki-67, PCNA, EGF, TGF-ȕ), PCR for p53 mutations. Results. Correlations between type of congenital malformation and histological variant of lung cancer were established. Morphologic analysis revealed five groups of patients: 80 cases / 73% / with different types of adenocarcinoma / papillary, solid adenocarcinoma with mucin productions, bronchioloalveolar adenocarcinoma / inside congenital cystic adenomatoid malformations of the lungs; 9 cases / 8% / - different grade squamous cell carcinoma incide bronchogenic cysts; - 9 cases / 8% / - different grade squamous cell carcinoma or adenocarcinoma incide simple hypoplasia; - 7 cases / 6.5% / different grade squamous cell carcinoma or adenocarcinoma inside congenital bronchiectasis; 5 cases / 4.5 % / adenocarcinoma or carcinoid inside intralobar sequestration. Precancer lesions in congenital malformation and lung cancer cells had similarities in expression of p53, bcl-2, c-myc, K-ras, CK19. p53 mutation were found in lung cancer cells, as well as in epithelial cell of precancer lesion in congenital malformation. Conclusion: obtained results may proof contribution of congenital malformations in the development of and lung cancer. PP3-128 VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) AND ITS RECEPTORS (VEGF-R1-Flt-1, VEGF-R2KDR/Flk-1) AS PROGNOSTIC INDICATORS IN SMALL CELL LUNG CARCINOMAS (SCLC) Evdokia Arkoumani1, Georgia Hardavella2, Yiotanna Dalavanga3, Petros Galanis4, Stavros Constantopoulos2, Dimitrios Stefanou1 1 Department of Pathology, University of Ioannina, Medical School, Ioannina, Greece 2 Department of Pneumonology, University of Ioannina, Medical School, Ioannina, Greece 3 Department of Anatomy, Histology and Embryology, University of Ioannina, Medical School, Ioannina, Greece 4 Laboratory of Epidemiology, University of Athens, School of Nursing, Athens, Greece Background: Tumor angiogenesis is a highly regulated process influenced by the host microenvironment and mediators. VEGF and its receptors (VEGF-R1-Flt-1, VEGF-R2-KDR/Flk-1) are good markers of vascular proliferation but their expression and relevance to tumor spread in SCLC is less clear. The present study aims to investigate angiogenesis and the expression of VEGF, VEGF-R1-Flt-1 and VEGF-R2-KDR/Flk-1 in SCLC and to establish their interrelationship and prognostic influence. Method: Immunohistochemical study of 50 SCLC was performed on paraffin sections with VEGF, VEGF-R1-Flt-1 and VEGF-R2KDR/Flk-1 antibodies (DBS California-Menarini Hellas). Mean vascular density (MVD) was assessed and rectospectively correlated with the clinical outcome. Results: A group of 50 patients (5 women, 45 men, mean age 64,3 years) included 25 with limited and 25 with extensive stage SCLC. 6%, 34% and 40% of tumor specimens presented high expression of VEGF, VEGF-R1 and VEGF-R2 respectively. 1/3 specimens presenting high VEGF expression, 18/20 having high VEGF-R2 expression and 12/17 with high VEGF-R1 expression corresponded clinically to metastasis. Mean survival for patients with high VEGF expression was 8,3 months and with low expression was 10,83 months. Mean survival corresponding to high VEGF-R1 and VEGF-R2 was 10,94 and 6,75 months respectively and to low was 10,35 and 9,41 months respectively. Conclusion: VEGF is expressed less than its two receptors (VEGF-R1-Flt-1, VEGF-

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R2-KDR/Flk-1) in SCLC. VEGF-R1 and VEGF-R2 expression are more closely correlated with metastasis than VEGF although the latter and VEGF-R2 seem to be positively correlated with poor patient outcome. PP3-129 EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR (VEGF) AND ITS RECEPTORS (VEGF-R1-Flt1, VEGF-R2-Flk1) IN NON SMALL CELL LUNG CARCINOMAS (NSCLC): CORRELATION WITH LYMPHANGIOGENESIS AND CLINICAL OUTCOME Georgia Hardavella1, Evdokia Arkoumani2, Yiotanna Dalavanga3, Stavros Constantopoulos1, Niki J. Agnantis2, Dimitrios Stefanou2 1 Department of Pneumonology, University of Ioannina, Medical School, Ioannina, Greece 2 Department of Pathology, University of Ioannina, Medical School, Ioannina, Greece 3 Department of Anatomy, Histology and Embryology, University of Ioannina, Medical School, Ioannina, Greece Background: Angiogenesis is an essential process in tumor growth and metastasis. VEGF and its receptors are considered to be good angiogenic markers but their correlation with lymphangiogenesis and patient outcome in NSCLC is yet to be defined. The present study aims to evaluate angiogenesis and the expression of VEGF, VEGF-R1-Flt1 and VEGF-R2-Flk1 in NSCLC, to correlate them with lymphangiogenesis (CD105 expression) and clinical outcome. Method: 96 NSCLC specimens were analyzed immunohistochemically using VEGF, VEFG-R1Flt1, VEGF-R2-Flk1 and CD105 antibodies (DBS CaliforniaMenarini Hellas). Mean vascular density (MVD) and intratumoral lymphatic microvessel density (ILMVD) were determined and rectospectively correlated with clinical outcome. Results: 13 women and 83 men (mean age 64,03 years) were included in our study. 15,6% were in stage I, 46,8% stage II, 40,4% stage III, 2,1% stage IV. 25% presented high VEGF expression, 40,6% high VEGF-R1-Flt1 and VEGF-R2-Flk1 expression and 87,5% high CD105 expression (ILMVD>25). 63 patients presented metastasis, 27% of which had high VEGF expression, 42,9% high VEGF-R1-Flt1 expression, 44,4% high VEGF-R2-Flk1 and 95,2% high ILMVD (p<0,05). ILMVD was significantly correlated with the stage of the disease (p<0,05) whilst angiogenic factors were not. Conclusion: Lymphangiogenesis proved to be of superior significance in being correlated with clinical outcome and predicting metastasis in NSCLC in comparison with VEGF and its two receptors (VEGF-R1-Flt1, VEGF-R2-Flk1) that seem to be of little significance. PP3-130 CLINICOPATHOLOGIC FEATURES AND PROGNOSTIC SIGNIFICANCE OF LUNG TUMORS WITH MIXED HISTOLOGIC PATTERN Ebru Cakir1, Funda Demirag1, Mehtap Aydin1, Ebru Unsal2 1 Ankara Ataturk Chest Disease and Chest Surgery Education and Research Hospital Department of Pathology, Turkey 2 Ankara Ataturk Chest Disease and Chest Surgery Education and Research Hospital Department of Chest Disease, Ankara, Turkey Background: Tumors with mixed histology are seen less often than the tumors with single histology in the lung. The prevalence is approximately in the range of 2-4%. The biologic behavior and clinicopathologic characteristics of these tumors have not been well described. We evaluated the clinicopathologic features and prognosis of lung tumors with mixed histology and compared them with the tumors that have single histology. Method: The study group consisted of 39 patients with a mixed histologic pattern and control group consisted of 41 patients with a single histology on the surgical specimen which were diagnosed between 1997-2007 at Ankara Ataturk Chest Disease and Chest

Surgery Education and Research Hospital.In the study group three types of tumors were identified: adenosquamous carcinoma, combined neuroendocrine tumors and biphasic tumors (epithelial + mesenchymal malignant components). The combined neuroendocrine tumors were further divided in small cell carcinoma (SCLC)+non-neuroendocrine carcinoma (NNEC), small cell carcinoma+large cell neuroendocrine carcinoma (LCNEC) and LCNEC +NNEC. Clinicopathologic characteristics, pTNM and survival rates were evaluated and compared with the population of resected lung tumors with single histology. Results: There were 20 adenosquamous carcinoma, 9 SCLC+ NNEC, 5 SCLC+LCNEC, 2 LCNEC+NNEC, 3 biphasic tumors as study group and 25 squamous cell carcinoma, 16 adenocarcinoma as control group. There was no significant difference for mean age, sex, smoking history, asbestos exposure and tumor size between study and control group (p>0.05). Among adenosquamous carcinomas advanced stage (IIIa or IIIb) was significantly more evident than the single histology group but survival rates were not different (p=0.328). Among combined neuroendocrine tumors advanced stages was more evident than adenosquamous carcinomas and single histology group (p=0.014). Survival was significantly worse for patients with combined neuroendocrine tumors than for patients with adenosquamous carcinoma and single histology group (p=0.0002). Median survival of biphasic tumors was 21 months (range 10-36). Univariate analysis revealed that tumor stage was significantly associated with the survival rate for all groups (p=0.016). Conclusion: Tumors with mixed histology are rarely seen in the lung. Among these tumors adenosquamous carcinoma and combined neuroendocrine tumors present more aggressive clinic behaviour than the tumors with single histology. PP3-131 STUDY OF THE POSSIBLE HORMESIS EFFECT IN AN EXPERIMENTAL PASSIVE TOBACCO MODEL IN THE EVOLUTION OF LUNG CANCER INDUCED BY URETHANE Patricia Maluf Cury, Ludmilla Nadir Santiago, Juliana De Camargo Fenley, Lucia Campanario Braga, José Antonio Cordeiro Faculdade de Medicina de São José do Rio Preto, Brazil Background: In a previous study, we verified an unexpected reduction of the number of pulmonary nodules in mice exposed to the cigarette smoke. One of the possibilities for such event would be a dose response effect called hormesis, which is represented by a J-shaped curve. The aim of the present study was to verify whether passive cigarette smoke might act by hormesis in an experimental model of lung cancer carcinogenesis. Method: 3mg/kg urethane was injected in male Balb-c mice, with 7 to 13 life weeks. After that, they were exposed to passive tobacco smoke twice a day, for five times a week, in a closed chamber. The animals were randomly divided into four groups. G0: only received urethane; G1: exposed to the smoke of three cigarettes per 10 minutes each time; G2: exposed to the smoke of three cigarettes per 10 minutes, twice; G3: exposed to the smoke of six cigarettes per 10 minutes, twice. Autopsy was performed after 16 weeks and the number of nodules and hyperplasia in lung was counted. Results: The statistics analysis showed that there was not significant difference when we compare the mean number of nodules and the hyperplasia between the studied groups (p=0,146 and p=0,513, respectively). Conclusion: The studied model did not show evidences of hormesis effect. Balb-C mice apparently are resistant to cigarette smoke exposure. Financial Support: FAPESP – Fundação de Amparo à Pesquisa do Estado de São Paulo.

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PP3-132 RESPIRATORY BRONCHIOLITIS – ASSOCIATED INTERSTITIAL LUNG DISEASE WITH FIBROSIS IN YOUNG PATIENTS PRESENTING WITH SPONTANEOUS PNEUMOTHORAX Elvira Stacher1, Stefan Scheidl2, Helmut Popper3 1 Insitute of Pathology, Medical University Graz, Austria 2 Department of Pulmonology, University Hospital, Medical University Graz, Austria 3 Institute of Pathology, Medical University Graz, Austria Background: Respiratory Bronchiolitis (RB) is a common incidental finding in lung specimens of smokers. It is defined as a bronchiolocentric accumulation of pigmented alveolar macrophages and is usually an incidental histological finding. In association with clinical symptoms such as shortness of breath, abnormalities of pulmonary function tests and/or CT scan findings the diagnosis of Respiratory Bronchiolitis-associated Interstistial Lung Disease (RB-ILD) can be considered. It is known that RB and RB-ILD are associated with a higher prevalence of spontaneous pneumothorax. Method: We retrospectively reviewed tissue of 14 cases of RB-ILD of young patients (age less than 35 years) presenting with spontaneous pneumothorax and requiring surgical pleurodesis. Among the study group there were 3 females and 11 males, the age ranged from 19-34 years with a mean age of 23,9 years. In 8 cases a CTscan was available. The H&E-stained sections were examined, and of areas with marked fibrosis also a Movat-stain were performed. Results: In 10 of 14 cases (1 female, 9 males) we observed a marked fibrosis, predominantly in subpleural and centrilobular areas. The Movat-stain allowed to discern early fibrosis with immature Collagen Type III and late scarring with mature collagen Type I. Conclusion: We think that Respiratory Bronchiolitis – Interstitial Lung Disease may progress to fibrotic lung disease since the occurrence of Collagene Type I is an irreversible event. This disease therefore deserves more attention and should be mentioned in the pathohistological report. PP3-133 ASBESTOS: MYTH, HISTORY, RELIGION Claudio Bianchi, Tommaso Bianchi Center for the Study of Environmental Cancer - Italian League against Cancer - Monfalcone, Italy BACKGROUND The most ancient evidences regarding the use of asbestos were traced in Finland and Sweden. In particular, asbestos was used in pottery in Eastern Finland from about 4000 B. C.. In the Mediterranean region the commerce of asbestos in the Classic Age is documented. In ancient era and in the Middle Ages the properties of asbestos were the subject of legends, myths, and theological comments. METHODS In the present study some Latin, Greek, and Italian literature texts were reviewed. RESULTS In the first century, the Latin encyclopaedist Pliny the Elder describes asbestos under various names in his Historia Naturalis. In one of these descriptions, asbestos is defined as a plant which grows in the desert of India. Pausanias, a Greek geographer of the second century, reports the presence of asbestos in the lamp placed in a temple in the Acropolis of Athens. An apocryphal gospel, the Protoevangelium of James, reports that different materials, including asbestos, were used in making the curtain of the Jewish Temple. After the Liber Pontificalis, a book in which the biographies of the first Popes are collected, an asbestos wick was placed on the candle in the middle of the Lateran baptistery in Rome. Asbestos is also quoted by the Christian Fathers of the Church, Basil the Great (339-379), and Augustin of Hippo (354-430). Basil states that the bodies of the three boys of Babylon, who after the tale of the Bible resisted to fire, had the nature of asbestos. An interesting description may be found in the Milione (Travels of Marco Polo) of the Italian traveller Marco Polo (1254-1324). In a region of Central Asia, Marco Polo saw towels that were placed in the fire

without burning. In addition, he reports that the Great Khan, the emperor of China, sent an asbestos towel as a gift to Rome; such towel had to be used “to wind the sudarium of our Lord”. CONCLUSIONS The review of the above texts shows that in the past the ‘positive’ features of asbestos were strongly emphasized. This agrees with the fact that asbestos was also used in medicine. Finally, the frequent association of the mineral with places of worship and with religious issues makes asbestos in some way sacred. PP3-134 MIXED SQUAMOUS CARCINOMA AND OSTEOSARCOMA (CARCINOSARCOMAS) OF THE LUNG HAVE A CGH MAPPING SIMILAR TO PRIMITIVE SQUAMOUS CARCINOMAS AND OSTEOSARCOMAS Javier Pardo1, Enrique De Alava2, Jesus Javier Sola1, Angel Panizo1, Gregorio Aisa1, Natalia Rodriguez-Spiteri1 1 Clinica Universitaria de Navarra. Universidad de Navarra, Spain 2 Centro de Investigacion del Cancer-IBMCC. Universidad de Salamanca, Spain Background: Carcinosarcomas (CS) are malignant tumors with a mixture of carcinomatous and differentiated sarcomatous elements. We selected from the surgical files of the Department of Pathology, University of Navarra three cases of CS composed of squamous carcinoma (SC) and osteosarcoma tissues. Methods: Clinical parameters were recorded. All cases were studied by light microscopy, immunohistochemistry and CGH technique. CGH assay was made in both carcinomatous ans osteosarcomatous areas, using as controls two lung squamous carcinomas and two lung metastatic osteosarcoma. Results: All patients were male and their age was 72, 43 and 58 years respectively. The size of the neoplasms was 7, 5.5 and 5 cm in maximum diameter. Only one patient had lymph node metastasis at presentation. All three patients died of their disease 4, 9 and 14 months after surgery. Pathologically, all cases had similar proportion of both squamous and osteosarcomatous structures by light microscopy. Immunoreactivity was positive for AE3AE1, CAM 5.2, CK-7, EMA, E-cadherine, p53 and CEA in carcinomatous areas and for vimentin and CD-68 in sarcomatous component. Foci of chondroid differentiation were positive for S100. Chromosomal imbalances using CGH were identified in all tumors from predominant epithelial and mesenchymal areas. Overall, a total of 55 copy number changes were detected with a median of 18 abnormalities per case (range: 12-26): 48 gains, 6 losses and one high level amplifications. We found similar chromosome alterations in carcinomatous areas and lung squamous carcinoma and in osteosarcomatous areas and metastatic osteosarcoma respectively. Conclusions: This study provided an accurate demonstration that the CGH alterations in osteosarcoma component in carcinosarcomas of the lung are similar to CGH changes of soft tissue osteosarcomas. Additional functional studies are needed to determine the biological and clinical significance of the CGH changes and theorize about the histogenesis of this unsual tumor. PP3-135 IS D2-40 A USEFUL MARKER DISTINGUISHING MALIGN MESOTHELIOMA, PULMONARY ADENOCARCINOMA AND BENIGN MESOTHELIAL PROLIFERATIONS? Hale Deniz1, Yasemin Kibar2, Muhammet Emin Guldur3, Kemal Bakir2 1 Gaziantep Pediatric Hospital, Turkey 2 University of Gaziantep, Medical Faculty, Department of Pathology, Turkey 3 Gaziantep Gynecologic and Obstetric Hospital, Gaziantep, Turkey

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Background: Pulmonary adenocarcinomas, malign mesotheliomas and sometimes benign mesothelial proliferations may show great resemblance histomorphologically. For this reason an immunodiagnostic panel is usually needed for differential diagnosis. Recently D2-40, a lymphatic marker, is suggested to be useful for mesothelial differentiation. Aim of this study is to compare D2-40 immunostaining in malignant mesothelioma, pulmonary carcinoma and benign mesothelial proliferations. Method: In this retrospective study, D2-40 immunostaining was investigated in 37 cases of malign mesothelioma, 36 cases of pulmonary adenocarcinoma and 31 cases of benign mesothelial proliferation. Diagnosis of malign mesothelioma were previously confirmed by a panel including calretinin, CK5/6, CEA. Results: Predominantly membranous immunoreactivity was observed in 51% of malignant mesothelioma and 55% of benign mesothelial proliferations. All of the 36 pulmonary adenocarcinomas were negative. These results were statistically significant (p<0.001). Conclusion: We believe that D2-40 may be helpful in differential diagnosis of malign mesothelioma from pleural involvement of pulmonary adenocarcinoma. PP3-136 PROGNOSTIC SIGNIFICANCE OF OSTEOPONTIN EXPRESSION IN MALIGNANT PLEURAL MESOTHELIOMA Susanna Cappia1, Luisella Righi1, Paolo Ceppi1, Marco Volante1, Elisa Bacillo1, Francesco Ardissone1, Dario Mirabelli2, Giorgio V. Scagliotti1, Mauro Papotti1 1 Department of Clinical & Biological Sciences, University of Turin at San Luigi Hospital, Turin, Italy 2 Cancer Epidemiology Unit, University of Turin at San Giovanni Battista Hospital, Turin, Italy Background: Malignant pleural mesothelioma (MPM) is an asbestos-related type of cancer with a median survival time of approximately 10 months. Long survivors are usually rare and currently no clear prognostic factor are strongly associated with such status. Recently, in asbestos exposed individuals, high osteopontin (OPN) serum levels were correlated to the presence of mesothelioma, but not with other asbestos-related conditions (pleural plaques or interstitial lung disease) and it has been proposed as a serum marker of mesothelioma in asbestos exposed subjects. Since OPN is a glycoprotein involved in cell-matrix interactions and cell signalling that was shown to be correlated with invasion, metastatic spread and decreased survival in several carcinomas and melanoma, the aim of this study was to analyse the prognostic role of OPN in MPM. Method: From a large database of 395 MPM cases diagnosed between 1989 and 2003 in a single institution, 26 cases with long survival (>24 months, mean 39.2 months) were extracted. The male/female ration was 16/10, median age 63.5, epithelioid versus biphasic histology 24 and 2/26, respectively, and extensive versus limited pleural involvement 17 and 9, respectively. None was surgically resected and treatment was talc pleurodesis associated with cisplatin-based chemotherapy All cases were matched to control cases with comparable clinical-pathological characteristics, except for a fatal outcome within 15 months (mean 8.9 months). OPN expression was detected by immunohistochemistry on archival biopsy specimens, the results quantified scoring extent and intensity of the immune reaction (scale 0-300), and the expression levels compared with survival and other parameters. Results: Tissue immunoreactive OPN was found in all cases, with a median score of 90 (range 5-285). Interestingly, OPN levels were significantly higher in short survivors compared to long surviving patients (Mann-Withney U-test, p<0.0001). At univariate analysis, Kaplan-Meier curves showed that patients with low (< median) OPN expression had longer survival compared to those with higher expression levels (38.7 vs 11.5 months, p=0.00001), while performance status, age, gender and extent of pleural involvement had no impact on the outcome. Conclusion: As

shown in several carcinomas, OPN expression levels in mesothelioma cells have a prognostic significance, being this protein probably involved in tumor progression and dissemination. It could represent a novel target for growth control strategies of this highly aggressive neoplasia.

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Hematopathology PP3-137 MEDIASTINAL B LARGE CELL LYMPHOMA A STUDY OF 11 CASES Atef Ben Abdelkader1, Mohamed Tahar Yacoubi1, Moncef Mokni1, Lilia Abid1, Adnen Laatiri2, Sadok Korbi3 1 Department Of Pathology, Tunisia 2 Department of Hematopathology, Tunisia 3 Depatment of Pathology, Tunisia Background : Mediastinal B large cell lymphomas is rare, distinguished from the other locations of lymphoma by its clinical and radiological features, its histopathological appearance and by a poor prognosis. The aim of this study is to specify the clinicopathological particularities, the management and the behaviour of this entity. Material and Methods : We studied 11 cases of mediastinal B large cell lymphomas diagnosed in the department of pathology during a period of 11 years (1993-2003). Clinical, radiological and biological informations provided from patient files. We used the index performance status (IPS) of the OMS to evaluate the clinical activity level of patients. Slides were reviewed to confirm the diagnosis . An immunohistochemistric study was performed to specify the proliferative index of tumors. The staging of tumors was evaluated according to the clinical staging system of Ann Arbor. Results : Our population was composed of 7 women and 4 men with a mean age of 25 years. The predominant presenting symptom was superior vena caval syndrome. The diagnosis was confirmed with an expression of CD20 by tumor cells in all cases. The IPS was low in 7 cases, which were stage IV of Ann Arbor system. Patients were treated chemotherapy associated to radiotherapy in three cases. Remission was noted in four cases and recurrence in three cases. No response was seen in four cases. Conclusion : Mediastinal B large cell lymphoma occurs preferentially in young females, most commonly revealed by superior vena caval syndrome. The diagnosis should be confirmed by histopathology and immunohistochemistry. A good response is a rule to radiation therapy and chemotherapy but recurrences are frequent. PP3-138 HETEROGENEITY OF MANTLE CELL LYMPHOMA Jirka Macak, Jana Smardova, Iva Kroupova, Barbora Ravcukova, Martin Klabusay, Leos Kren Department of Pathology, University Hospital Brno, Czech Republic Background: the aim of this study is to investigate the heterogeneity of mantle cell lymphoma (MCL) in morphology, immunohistochemistry, FISH and molecular biology. Material and Methods: in the archive of Department of Pathology University Hospital Brno we found 27 cases of MCL in lymph nodes. Classical histology, immunohistochemistry, FISH method and from native tissues FASAY (Functional Analysis of Separated Alleles in Yeast) were performed. Results: in series of 27 cases of MCL were 17 men and 10 women. Most patients were in age of 61-80 years. Morphologically, diffuse infiltration with medium size lymfoid cells was found. Mitotic rate (number of mitoses/10HPF) in first category (0-5 mitoses) in 13 cases; second category (6-10 mitoses) in 8 cases; third category (11-20 mitoses) in 4 cases; fourth category (21-30 mitoses) in two cases. Highly characteristic marker for MCL, hyalinne deposits, was recognized in 20 cases. FISH for t(11;14) was performed in all cases and 51-100% of tumour cells were positive. Immunohistochemistry revealed positivity with marker CD 5 in 19/27 cases (30% cases were negative); CD 20 in 27/27; CD 43 in 23/27; cyclinD-1 in 19/27 (30% cases were negative); bcl-2 in 26/27; p53 in 22/27. The proliferation marker Ki-67 was detected in approx. 20% of cells in 16 cases and 30-60% of cells were found in 11 cases. For status p53 by FASAY method, 12 native tumour tissues were examined. In the two of them, abberation in

the p53 gene was revealed and missense p53 mutations R282W and C288S were detected by DNA sequencing. Conclusions: our results show that MCL was uniform in morphology and in type of infiltration, which was in all cases diffuse. Very useful marker for diagnosis seems to be hyaline deposits stcattered in the lymphoma tissue. Number of mitoses enable us include 6 cases of the blastoid variant of MCL. In immunohistochemicstry, CD 5 and cyclinD-1 markers were negative in 30 cases. Abberation of p53 gene is rare in MCL. This work was supported by grant NR/9305-3 of The IGA MZ CR. PP3-139 PROLIFERATION MARKERS IN ZAP70+ AND ZAP70- B CELL CHRONIC LYMPHOCYTIC LEUKAEMIA Petra Mandakova, Katerina Kamaradova, Hana Kortankova, Jana Siftancova, Eliska Stranska, Vit Campr Department of Pathology and Molecular Medicine, Charles University in Prague, 2nd Medical Faculty, Prague, Czech Republic Background: B cell chronic lymphocytic leukaemia (B-CLL) is a low grade lymphoproliferative disorder. Generally, most patients are asymptomatic with a long survival, but there exists a subset of cases with a more aggressive course. The expression of protein ZAP70, often associated with non mutated status of IgVH genes, and expression of CD38 molecule divide B-CLL into prognostically relevant subgroups. In this study we correlated these subgroups with the expression of various proliferation markers (CD71, proliferation index (PI) and Ki67). Methods: We analyzed 50 samples from 40 patients with B-CLL/SLL (bone marrow, lymph nodes, peripheral blood and tonsils). The expression of ZAP70 and CD38 molecules on CD19+CD5+ tumor cells were assessed in cell suspensions by means of flow cytometry, as well as the expression of CD71 (transferrin receptor). The PI was defined as a number of cells in S and G2/M phase of cell cycle determined by flow cytometry analysis. Ki67 was detected immunohistochemically in formalin-fixed paraffinembedded material if available. Results: 32 samples from 26 patients were ZAP70+ and 17 samples from 14 patients were ZAP70-. The expression of CD38 subdivided samples into ZAP70+CD38+ (30 %), ZAP70+CD38- (35 %), ZAP70-CD38+ (4 %) and ZAP70-CD38- (31 %). The higher proportion of sample infiltration, higher CD71 expression and higher proliferation was found in ZAP70+ patients compared to B-CLL without expression of ZAP70. The information on status of CD38 brought no significant difference. Conclusion: B-CLL is clinically as well as biologically a heterogenous disorder. This fact is reflected by description of various markers which divide the disease to subcategories with different prognosis. In the investigated set of patients we showed that the ZAP70 positivity of B-CLL cells is followed by a higher expression of proliferation markers and implicates a heavier tumor infiltration of the tissue. We conclude that the expression of ZAP70 is associated with a higher proliferative activity and thus can be a good marker of more aggressive disease with worse prognosis. This study was supported by Research Project FNM MZO 00064203 n. 6704. PP3-140 FIRST EXPERIENCES WITH DETECTION OF JAK2 MUTATION IN PARAFFIN EMBEDDED BONE MARROW BIOPSIES OF POLYCYTHEMIA VERA PATIENTS Juraj Marcinek1, Luká Plank1, Tatiana Burjanivová1, T. Balhárek1, Peter Szépe1, Zora Lasabová2 1 Department of Pathology and National Consultation Centre of Hematopathology Jessenius Faculty of Medicine, Comenius University and Faculty Hospital in Martin, Slovakia 2 Department of Molecular Biology, Jessenius Faculty of Medicine, Comenius University in Martin, Slovakia

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Background: Point JAK2 mutation (V617F) was recently identified in association with Ph1- myeloproliferative diseases (MPD) in almost all polycythemia vera (PV) patients and in cca 50% of patients with essential thrombocythemia and chronic idiopathic myelofibrosis. The mutation shows association with typical laboratory and clinical findings, while its prognostic significance remains uncertain. It might be identified by direct DNA sequencing, allele-specific PCR (ASPCR), real-time PCR, DNA-melting curve analysis, etc. In majority of published papers fresh peripheral blood or bone marrow (BM) cell analysis was used, but only limited experiences based on retrospective analyses of BM biopsies are known. The aim of this study was to prove the possibility of DNA analysis obtained from the regular BM biopsies with the aim to implement JAK2 mutation detection into the routine praxis of our hematopathology center. Methods: The paraffin-embedded BM biopsies of PV patients diagnosed in the period January 2004 – March 2007 were selected from the files and reevaluated histologically and clinically. BM sections were deparaffinized in graded alcohols and xylene and digested with proteinase K over two nights. DNA was extracted using the Wizard® genomic DNA purification kit (Promega). Detection of the V617F mutation was performed by allele-specific hot start multiplex PCR with one common reverse and two separate forward primers. In the presence of the V617F JAK2 mutation two PCR products of different sizes were amplified, while in the presence of wild-type DNA only one fragment is amplified. Results: BM biopsies of 53 patients fulfilled WHO criterias of PV diagnosis established by BM biopsy and/or by hematologist. In addition, the clinicians were subsequently asked to report the follow up. At the time of abstract submission, results of JAK2 detection of 19 patients were available. BM biopsy showed typical PV morphology in 18 cases and MPD NOS in 1 case and the diagnosis was confirmed also by follow up in 13 of them. JAK2 mutation was detected in 16 of 19 patients and in all of 12 cases sharing both typical morphology and clinical behaviour of PV. Conclusion: Although the importance of JAK2 detection for MPD typing is limited, its diagnostic implementation is usefull and its possible therapeutic significance might not be excluded. Our preliminary results confirmed a high sensitivity of JAK2 detection by using routine BM biopsy material. Further analyses will follow, incl. the atempts to identify its homozygous and heterozygous forms. Supported by Grant UK/84/2007. PP3-141 OPPORTUNISTIC INFECTIONS IN HIV+/AIDS PATIENTS – COMPARATIVE AUTOPSY STUDY Florica Staniceanu1, Sabina Zurac1, Gianina Micu1, Alexandra Bastian1, Luciana Nichita2, Cristiana Popp1, Andrei Murariu1, Claudiu Socoliuc1, Liana Tudorica1, Adrian Streinu-Cercel3 1 Colentina University Hospital, Bucharest, Romania 2 Carol Davila University of Medicine and Pharmacy, Bucharest, Romania 3 Matei Bals Institute, Bucharest, Romania Background: Patients with HIV/AIDS have an important immunodepression, which makes them susceptible for opportunistic infections which are, in most cases, the main cause of death. Material and method: We made a comparative study of autopsy findings regarding infectious pathology including all AIDS patients submitted to autopsy in our department as follows: group A – patients HIV positive autopsied between 1997-2001 (27 cases) and group B – 19 cases between 2002-2006. Results: The cause of death in our two groups varied as it follows: • group A: - opportunistic infections (single or combined) - 22 cases (7 cases of each tuberculosis and systemic candidosis, 4 cases of pneumocystosis, 2 cases of cryptococcosis, 1 case of each Cytomegalovirus infection and systemic aspergillosis). nonspecific infections - 4 cases - other causes – 1 case (nonhodgkinian malignant lymphoma) • group B: - opportunistic infections (single or combined) – 13 cases (3 cases of

tuberculosis, 2 cases of each pneumocystosis, toxoplasmosis, progressive multifocal leukoencephalopathy and Cytomegalovirus infection, 1 case of coccidioidomycosis and 1 case of systemic aspergillosis. - nonspecific infections – 4 cases other causes – 2 cases (one cerebral malignant lymphoma and one myocardial infarction). There is a definite change in the spectrum of lethal opportunistic infections in AIDS related deaths in Romania - a lower rate of tuberculosis in group B comparing with group A, disappearing of lethal systemic candidosis and occurence of other type of opportunistic infections: toxoplasmosis and progressive multifocal leukoencephalopathy due to JC virus infection. The most probable cause of this trend is high active antiretroviral therapy (HAART) which assures a tolerable level of immunity for these patients. The majority of the group B patients who died by opportunistic infections were recently found HIV+ (1-3 months) and HAART was either not used or very recently administrated. Conclusion: Opportunistic infections are an important health problem and a frequent cause of death in HIV positive patients. This study provides informations about changes occurred in the main causes of AIDS related deaths, data that may be useful for improving the medical management of the HIV+/AIDS patients. PP3-142 BONE MARROW (BM) PATHOLOGY CONTRIBUTES TO “IN SITU” UNDERSTANDING OF CHRONIC LYMPHOCYTIC LEUKEMIA (CLL) IMMUNOPATHOGENESIS: ANALYSIS OF A SERIES OF 92 PATIENTS Theodora Papadaki1, Kostas Stamatopoulos2, Niki Stavroyianni2, Maria Fameli1, Anastasia Athanasiadou2, Chrysavgi Lalayanni2, Anastasia Hadzidimitriou2, Ioanna Athanasiadou2, Evi Puliou1, Achilles Anagnostopoulos2, Athanasios Fassas2, Dimitra Anagnostou1 1 Evangelismos Hospital, Greece 2 G. Papanicolaou Hospital, Greece We conducted a detailed analysis of BM biopsy (BMB) samples from a series of 92 CLL patients and explored associations with clinical features and outcome, CD38 expression, and IGHV mutation status. Patient group: M:F=54:38, median age: 66 years, Binet stage-A/B/C: 76/12/4. Peripheral blood immunophenotype: (1) CD38+: 35/92 cases (2) IgG+: 19/92 cases. Mutated/unmutated IGHV genes (M-IGHV/A-IGHV): 59/33 cases. Formalin-fixed, decalcified, paraffin-embedded sections of BMBs were examined: (1) morphologically (H&E); (2) immunohistochemically (ABC technique). The following antibodies were used: CD20, CD79a, CD3, BCL-2, CD5, CD23, SIg/CIg (kappa/lambda/mu/delta/gamma/alpha), CD27, myeloperoxidase, PGM1 (CD68), glycophorin-C, CD61. All cases showed neoplastic lymphocytic infiltration (20-95% of BM cellularity). In all cases, neoplastic lymphocytes expressed CD20/CD79a/CD5/CD23. In the vast majority of cases, the neoplastic population consisted almost exclusively of small lymphocytes. Medium- or large-sized cells were admixed in 4 and 2 cases, respectively. A moderate number of monocytoid-like cells with clear cytoplasm were identified in 3 cases. Finally, 5 cases exhibited lymphoplasmocytoid differentiation in a small proportion of the neoplastic population. Three patterns of neoplastic lymphocytic infiltration were identified: (1) interstitial: 41 cases; (2) nodular/nodular+interstitial: 31 cases; (3) diffuse: 20 cases. Moderate-to-significant reduction of the granulocytic series was observed in 47/92 cases. The remaining cases showed hyperplastic granulocytic series with a left shift and, occasionally, dysplastic changes of mature forms. Hyperplasia of the erythroid and megakaryocytic series -often with dyserythropoiesis and dysmegakaryopoiesis- in a context of normal/decreased hemoglobin levels and platelet counts was observed in 52/92 and 72/92 cases, respectively; the remaining cases showed moderate-to-significant reduction of either series. Significant correlations were identified between: (i) nodular

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infiltration and M-IGHV genes (p<0.001); (ii) diffuse infiltration and shorter time-to-progression (p=0.05). In conclusion, BMB examination: (1) permits identification of the rare CLL subtype with plasmocytoid differentiation (WHO); (2) allows evaluation the BM stroma and the hematopoietic marrow and provides important evidence for CLL-associated hematopoietic autoimmunity. Finally, the present study indicates that the favorable prognosis of the nodular pattern of infiltration in CLL may be interpreted in the context of its association with M-IGHV genes. PP3-143 A CASE OF V617F(+) MYELODYSPLASTIC / MYELOPROLIFERATIVE DISORDER Ozden Ozer1, Emre Eskazan2, Huseyin Bekoz3, Kamil Peker1, Sirin Yuksel4, Ender Altiok4, Burhan Ferhanoglu2 1 Istanbul Patoloji Grubu, Turkey 2 Cerrahpasa Tıp Fakültesi, Hematoloji Departmanı, Turkey 3 Onurlu Saglık Hizmetleri, Turkey 4 Acıbadem Genetik Tanı Merkezi, Turkey Background: Mutations involving the JAK-2 tyrosine kinase have been described in a great majority of Polycytemia Vera(PV)and to a lesser extend in Essential Thrombocytemia(ET) and Chronic Idiopathic Myelofibrosis(CIMF). Myelodysplastic syndromes( remained mostly (-) for JAK-2 mutation except a unique subgroup with persistant presence of anemia, ringed sideroblasts and thrombocytosis. While some of these were referred as Refractory anemia with ring sideroblasts –Thrombocytosis (RARS-T ) in the literature some were classified as Myeloproliferative / Myelodysplastic Disorder– unclassifiable (MPD/MDS–u). Here we are reporting a patient with a V617FJAK(+) MPD/MDS-u, characterised by anemia, ring sideroblasts, thrombocytosis,granulocytosis and multilineage dysplasia.Material&Methods: A 53 year old male presented with recurrent gastrointestinal bleeding. He was found to have anemia,thrombocytosis, granulocytosis and hepatosplenomegaly. In contrast to an iron deficiency anemia, expected in a setting of recurrent bleeding, he had macrocytic anemia. Upon completion of his acute management, the patient underwent a bone marrow biopsy in the outpatient setting. His bone marrow biopsy showed hypercellularity(%100) due to granulocytic and megakaryocytic expansion. Erythroid precursors were decreased and there was multilineage dysplasia. Numerous ring sideroblasts were identifed on the Prussion blue stain. The case was classified as MPD/MDS-u and a correlation with JAK-2 mutation was suggested by the pathologist. Real time PCR with melting curve analysis was performed for V617FJAK-2 on genomic DNA isolated from peripheral blood. Results : Our case characterised by anemia, thrombocytosis, ring sideroblasts, leucocytosis and multilineage dysplasia, classified as MPD /MDS–u revealed V617FJAK-2. Discussion : Detection of JAK-2 mutation with different diseases raises the possibility of presence of additional mutations determining the final disease phenotype. V617F mutation is known to render JAK-2 constitutively active resulting in abnormalmyeloproliferation. Therefore, in the myeloproliferative disease PV, JAK-2 mutation may be the sole genetic anomaly, correlating with the promising results obtained with JAK-2 inhibitors. However, in the MPD/MDS category, sole JAK-2 inhibition may not be sufficient in controlling the dysplastic component. Although, effects of JAK-2 inhibitors remain to be tested in clinical practice,further studies are needed to uncover cooperating mutations in this category.

PP3-144 c-MYC GENE TRANSLOCATION AND INCREASED COPY NUMBER PREDICT POOR PROGNOSIS IN ADULT DIFFUSE LARGE B CELL LYMPHOMA (DLBCL), ESPECIALLY IN GERMINAL CENTER-LIKE B CELL (GCB) TYPE Yoon Kyung Jeon1, Sun Och Yoon1, Young A Kim2, Ji Eun Kim2, Chul Woo Kim1 1 Department of Pathology, Seoul National University College of Medicine, Seoul, Korea 2 Department of Pathology, Seoul National University Boramae Hospital, Seoul, Korea Diffuse large B-cell lymphoma (DLBCL) is a heterogeneous disease showing variable clinical presentations, histology, and molecular and cytogenetic features. In this study, c-MYC, BCL2, and BCL6 gene translocation and gene copy number change were comprehensively investigated in adult DLBCLs using interphase fluorescence in situ hybridization (FISH) analysis in formalinfixed paraffin-embedded tissues, and their clinicopathologic features and prognostic implications were analyzed, especially in the context of germinal center B-cell like (GCB) and non-GCB type of DLBCLs based on immunohistochemistry. C-MYC translocation was observed in 9% (14 of 156), and increased gene copy number (ICN) was in 7.1% (11 of 156) of DLBCLs. Of the 14 cases with c-MYC translocation, 12 had IgH gene as a fusion partner. C-MYC translocation was significantly more common in GCB type DLBCLs than non-GCB type (22% [9 of 41] in GCB vs 4.9% [5 of 103] in non-GCB, p = 0.004). C-MYC translocation or ICN was not associated with Ki-67 labeling index. However, c-MYC translocation or ICN exhibited association with decreased BAX expression and increased BIM expression (p = 0.022 and 0.001, respectively). Clinically, cMYC translocation was closely related with advanced IPI group (p = 0.005). In univariate survival analysis, c-MYC aberration including translocation or ICN was associated with shorter overall survival (p = 0.012) in adult DLBCLs, especially within GCB type (p = 0.030). BCL2/ IgH translocation in DLBCLs was only observed in 3.4% (5 of 145) in our cohort, which was much lower in comparison with the reports of Western countries. BCL2 ICN was detected in 11.7% (17 of 145), more frequently in nonGCB type DLBCLs than GCB type (16% [15 of 94] in non-GCB vs 2.5% [1 of 40] in GCB, p = 0.038). BCL2 ICN and/or BCL2/IgH translocation was well correlated with BCL2 protein expression (p = 0.031). BCL2 aberration such as ICN or translocation tended to show adverse effect on the overall survival of patients, but without statistical significance. BCL6 translocation was observed in 17.5% (24 of 137), which was most common gene aberration in our study. However, BCL6 translocation had no influence on the prognosis of DLBCL patients. In conclusion, we suggest that the evaluation of c-MYC gene status including translocation and increased gene copy number and the analysis of the results in the context of DLBCL phenotype might be important to predict the prognosis of DLBCL patient and determine the therapeutic strategies. PP3-145 A SIX-MONTH ART INTERRUPTION IN HIV-INFECTED PATIENTS IMPROVES ADIPOSE TISSUE MORPHOLOGY AND GENE EXPRESSION Cervera Pascale Hopital St Antoine, France To determine the ability of adipose tissue to recover after sixmonth ART interruption, subcutaneous abdominal adipose tissue of forty well-controlled HIV-patients with thirty three completed the study was studied at inclusion and after 6 months. Adipose tissue morphology, mithochondrial DNA and gene expression were evaluated. At M6, clinical parameters were not significantly modified, but inflammation markedly improved with a decreased in numbers of macrophages, and of adipocytes or inflammatory

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cells stained with TNFĮ and IL6. The expression of adipogenic factor PPAR gamma and of markers associated with mitochondrial function and biogenesis, COX2 and PGC1Į, was improved. Mitochondrial alterations were improved with decreased COX4 mRNA and increased mtDNA and COX2/C0X4 ratio. These data clearly outline the delerious impact of the thymidine analogues on adipose tissue. Even if no modification was clinically noticed, a 6-month treatment interruption allows a partial recovery of adipose tissue alterations. PP3-146 SIMIAN VIRUS 40 IN KOREAN PATIENTS WITH NONHODGKIN’S LYMPHOMAS Young A Kim1, Ji-Eun Kim1, Yoon-Kyung Jeon2, Chul-Woo Kim2 1 Department of Pathology, Seoul National University Boramae Hospital, Korea 2 Department of Pathology, Seoul National Univertiy Hospital, Korea DNA sequencies coding for simian virus 40 (SV40) large T antigen have been detected in various human tumors, including non-Hodgkin's lymphomas (NHLs). But the relationship between SV40 and NHLs was controversial and needs to be studied. 89 cases of NHLs and 20 control lymph node tissues were tested for SV40 sequences using PCR amplification and sequencing. Viral sequences were present in two (2.2%) of 89 NHLs and were absent in 20 control tissues. Two SV40 positive cases were both intestinal diffuse large B cell lymphomas, and immunohistochemical staining for SV40 large T antigen was completely negative. These results suggest that SV40 is unlikely to have played a pathogenetic role in most of lymphoproliferative disorders in the Korean population. PP3-147 UNUSUAL SITES OF B-CLL INVOLVEMENT: BIOPSY DIAGNOSIS OF B-CLL INFILTRATES PRESENTING IN EXTRANODAL AND EXTRAMEDULLARY TISSUES Tomas Balharek, Lukas Plank, Peter Szepe, Juraj Marcinek, Zuzana Kviatkovska, Martina Barthova Department of Pathology and National Consultation Centre of Haematopathology, Comenius University, Jessenius Faculty of Medicine and Faculty Hospital, Martin, Slovakia Background: Majority of B-CLL/SLL cases with the diagnosis established by a biopsy is based on examination of lymph node (LN) or bone marrow (BM) specimens using WHO classification. Less commonly the patients show extranodal and/or extramedullary B-CLL manifestation, requiring an endoscopic or surgical intervention and biopsy verification of the tumor infiltrate. The aim of this study was to evaluate the possibilities of an appropriate B-CLL/SLL biopsy diagnosis in such cases. Methods: We reviewed records of 716 biopsies of B-CLL/SLL patients performed in the interval 01/2004 – 03/2007 and selected those presenting in other than LN and/or BM localisations. All the cases were reevaluated in sections stained with HE and Giemsa staining and by immunohistochemical detection of CD20, CD23, CD43, CD3 or CD45RO and Ig light chain. In addition, the biopsy register was analyzed to identify all other previous or subsequent biopsies of the patients. Results: From all reviewed biopsies only 15 cases (2,1%) represented infiltrates of other than LN or BM tissues, 3 of them represented a high grade transformation of B-CLL. In 5 patients the disease was diagnosed by tonsillectomy and in 10 cases by involvement of nonlymphoid organs: upper respiratory tract (n=3), trachea (n=1), thyroid gland (n=1), skin (n=3) and soft tissues (n=2). BCLL/SLL diagnosis was confirmed: a.) by typical morphology and phenotype (n=12), incl. 2 cases of CLL with coincidental diffuse large B-cell lymphoma and classical Hodgkin lymphoma, resp. b.) by previous or subsequent BM (n=6) or LN biopsy (n=3) confirming the B-CLL diagnosis, incl. also cases problematic at the actual time of biopsy. These cases included: 2 limited and

partially necrotic specimens insufficient for the complex analysis, especially in relation to confirmation CD23 positivity and 1 skin biopsy of a patient with B-CLL showing diffuse large B-cell lymphoma without residual CLL infiltration. Conclusion: The biopsy examination of B-CLL infiltrates appearing in the extranodal localisation seems to represent a rare event. These cases may cause diagnostic and differential diagnostic problems, while it might be difficult to follow the WHO criteria in limited material from unusual sites of involvement. Also the evaluation of high grade transformation might be problematic. In such cases the diagnosis should be supported by a complex clinical evaluation and/or by a BM and LN biopsy. Supported by Grants of Slovak Ministry of Health Nr. 2005/12-MFN-04, VEGA Nr. 1/4285/07 and Comenius University Nr. UK/83/2007. PP3-148 PROGNOSTIC SIGNIFICANCE OF Bcl-2, P53 PROTEIN EXPRESSION AND Ki67 PROLIFERATIVE INDEX IN DIFFUSE LARGE B-CELL LYMPHOMA Betul Bolat Kucukzeybek1, Sadi Bener2, Aylin Orgen Calli2, Tugba Dogruluk Paksoy3 1 Department of Pathology, izmir Training Research Hospital, Turkey 2 Department of Pathology, Izmir Ataturk Training and Research Hospital, Turkey 3 Department of Pathology, KahramanmarasYenisehir Government Hospital, Turkey Background: Diffuse Large B-Cell Lymphoma (DLBCL) is a high grade neoplasm which has a heterogeneous properties in clinical, morfological, immunophenotypical aspects. DLBCL exhibits varies markedly in response to treatment and prognosis. Clinicopathological and biological factors must be investigated and evaluated to determinate the prognose of the disease. In our study we searched effects of p53 (tumor supressor gene) which is a cell cycle regulator, bcl-2 oncoprotein which is an inhibitor of apoptosis and a cell proliferation marker Ki-67, on prognosis and relations with clinical parameters. Method: Total of thirty five patients who had been diagnosed as nodally located DLBCL at the Izmir Ataturk Training and Research Hospital between January 1999-June 2006 were included in this study. The Ann Arbor classification used to detemine the stage of patients. Patients were evaluated according to the age, sex, stage, B symptoms, extranodal involvement, lactate dehydrogenase (LDH) level and also immunohistochemically; p53 expression, bcl-2 oncoprotein expression and Ki-67 proliferation index. Statistical anlyses were performed using Chi-square, Fishers Exact and Logrank test. Results: High Bcl-2 expression was identified in 9 patients (%25.7), high P53 expression was identified in 10 patients (%28.6) and high Ki67 expression was observed in 23 patients (%65.7). There was no statistical significant corelation between the p53, bcl-2 expression, Ki 67 proliferation index and age, sex, stage, B symptoms, extranodal involvement, LDH level, overall survival ( p>0.05). There was no statistical significant relationship between p53, bcl-2, Ki 67 expression and prognosis ( p>0.05). There wasn’t a statistical significant relationship between overall survival and age, sex, stage, B symptoms, extranodal involvement, LDH level (p>0.05). Conclusion: As a result in our study we found that bcl-2 and p53 protein expressions, Ki67 proliferation index have no effect on overall survival of patients with DLBCL. The prognostic importance of p53, bcl-2 protein expression, Ki 67 proliferation index in DLBCL which has a biological and clinical heterogenity, can be understood in large series studies which have subclasses and with immunohistochemical markers which have optimal cut off values.

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PP3-149 SPLENIC MARGINAL ZONE LYMPHOMA WITH ABERRANT CD5 EXPRESSION: REPORT OF A CASE Fulya Oz Puyan1, Ufuk Usta1, Cigdem Ozdemir1, Burhan Turgut2, Erman Ozturk2, A. Kemal Kutlu1 1 Trakya University Medical Faculty Department of Pathology, Edirne, Turkey 2 Trakya University Medical Faculty Department of Hematology, Edirne, Turkey Splenic marginal zone lymphomas (SMZLs) are low-grade B-cell lymphomas that characteristically lack CD5 expression. However, rare cases of marginal zone lymphomas have been reported with CD5 coexpression which show an aggressive clinical course than the CD5- negative SMZLs. We represent here a 44-year-old male with splenomegaly and leucocytosis in laboratory findings. Peripheral blood smears showed monocytoid B cells with villous projections. Flow cytometry of the peripheral blood revealed CD19, CD20, CD22 and FMC7 positivity without CD5, CD23, CD10, CD38, and CD103 positive expression. The spleen histology showed a distinctive pattern of white pulp infiltration by abnormal B lymphocytes with marginal-zone differentiation. Immunohistochemical findings of the spleen were consistent with flow cytometric analysis of the peripheral blood. Mononuclear cells showed CD20 positivity without CD5 and CD23 expression, However bone marrow biopsy revealed an interstitial and nodular lymphoid infiltration with strong CD5 immunopositivity. Clinicians and pathologists must be aware of this unusual aberrant CD5 expression in SMZLs on bone marrow infiltrate in order to reach a correct diagnosis. PP3-150 CD 30 POSITIVE T- CELL LYMPHOPROLIFERATIVE DISORDER: A CASE REPORT Fulya Oz Puyan1, Emre Tekgunduz2, Aysegul Ilgili1, Gulsum Pamuk2, Omer Yalcin1, Oner Dogan3 1 Trakya University Medical Faculty Department of Pathology, Edirne, Turkey 2 Trakya University Medical Faculty Department of Hematology, Edirne, Turkey 3 Istanbul University Istanbul Medical Faculty Department of Pathology, Turkey Introduction: CD30-positive T-cell lymphoproliferative disorders (CD30+ T-LPD) of the skin include a heterogeneous group of diseases which differ in their clinical and histological presentations. Lymphomatoid papulosis (LyP), primary cutaneous anaplastic lymphoma (C-ALCL) and borderline lesions constitute CD30+ T-LPD. The diagnosis of C-ALCL and LyP is very difficult as many of them have overlapping clinical, histologic, and immunophenotypic features. Most patients within this spectrum of disease have an excellent prognosis with 5-yearsurvival rates of 100% for LyP and %90 for C-ALCL. Extracutaneus spread and regional lymph node involvement may be seen on patients with multifocal tumor lesions. LyP is generally managed by observation whereas C-ALCL is treated with spot radiation or surgical excision with systemic chemotherapy reserved for cases with large tumor size or extracutaneous involvement. Therefore it is necessary to differentiate these types of disorders from each other. We report here a case of CD30+ T-LPD with multiple skin nodules and no signs of generalization, lymph node or internal organ involvement. Case Report: A 63-year old male represented with increasing number of skin papules and nodules on his belly, arm, back and subcostal region. The largest one 4x3 cm in size enlarged rapidly and became ulcerated. The patient had a history of bladder carcinoma in 1998 without any postoperative chemoor radiotherapy. The physical examination revealed no lymphadenopathy or organomegaly. Initial biopsy showed a subepidermal dense infiltrate of large lymphocytes with irregular nucleus and abundant clear-eosinophilic cytoplasm intermixed

with a few reactive cells. Immunohistochemistry revealed strong and widespread CD30 positivity with expression of T-cell associated antigens. The clinical, histologic, and immunohistochemical findings pointed to a borderline lesion of CD30+ T-LPD. The patient was treated with one dose of CHOP protocol. After the 3 month-follow-up period a rapidly regress of all the nodular lesions was noticed. Conclusion: This report adds to the clinical and morphologic spectrum of the CD30+ T-LPD with uncertain progress and treatment. Accurate recognition of this lymphoma and follow up is needed. PP3-151 CYTOGENETIC ABERRATIONS OF CHROMOSOME 1 IN B-CELL NON HODGKIN LYMPHOMAS: CORRELATIONS WITH DISEASE SUBTYPE Govind Bhagat, Shivakumar Subramaniyam, Subhadra Nandula, Vundavalli Murty Columbia University, New York, USA Background: B-cell non Hodgkin lymphomas (B-NHL) comprise a variety of distinct entities that are associated with characteristic chromosome (chr) abnormalities. Many B-NHL acquire additional, recurrent cytogenetic aberrations during disease evolution. Abnormalities of chr 1 are considered a frequent secondary event in the cytogenetic progression of certain B-NHL. We undertook this study to analyze the location and frequency of chr 1 abnormalities in a large series of B-NHL to determine differences and common regions of alterations in distinct types of B-NHL. Methods: G-band karyotypes with clonal abnormalities, obtained from tissue submitted for cytogenetic analysis for suspected B-NHL, were reviewed to detect chr 1 abnormalities. All numerical and structural chr 1 aberrations were mapped on an ideogram to identify specific regions of gains and losses and sites of translocations. The B-NHL were classified according to current WHO criteria. Results: Chr 1 abnormalities were detected in 110/275 (40%) cases comprising 103 (37.4%) B-NHL; 58 diffuse large B-cell lymphomas (DLBCL), 21 follicular lymphomas (FL), 13 Burkitt lymphomas (BL), 5 mantle cell lymphomas (MCL), 5 marginal zone B-cell lymphoma (MZBCL), 1 chronic lymphocytic leukemia (CLL), and 7 (2.5%) reactive lymphoid hyperplasias (RLH). Isolated chr 1 abnormalities were detected in 2 B-NHL. Intra or interchromosomal translocations were found in 77/103 (74.7%) B-NHL and 4/7 (57%) RLH. Gains of chr 1 resulting from duplications, triplications, and homogeneously staining regions were observed in 47.6%, 6.8%, and 1.9% B-NHL, respectively and in 37.5% RLH. HSR were only detected in DLBCL. Deletions were identified in 33/103 (32%) B-NHL and 1/7 (14.3%) RLH. Gains were restricted to 1q and losses predominantly involved 1p, only 9 (8.7%) B-NHL showed 1q losses. The single CLL had a 1q translocation. The majority of rearrangement breaks were localized to chr 1q21.1 in all B-NHL subtypes. However, differences in common regions of gain/amplification (1q21-23 in BL and MZBCL and 1q21-31 in FL and DLBCL) and deletion (1p32-34 in MCL and 1p36 in FL and DLBCL) were noted. Conclusions: In addition to a high frequency of secondary chr 1 alterations, we detected primary chr 1 abnormalities in B-NHL and RLH. Common and different cytogenetic aberrations of chr 1 in B-NHL subsets indicate shared as well as distinct pathways of lymphoma progression. Elucidation of genes whose expression levels are altered due to chr 1 copy number changes could help identify tumor suppressor genes and protooncogenes of prognostic relevance. PP3-152 PRIMARY FOLLICULAR LYMPHOMA OF THE GALLBLADDER Leyla Abid, Olfa Ismail, Aida Ayadi, Faouzi El Mezni Department of Pathology, AMami Hospital, 2080 Ariana, Tunisia

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Primary lymphomas of the gallbladder are extremely rare, and follicular lymphoma has been reported only twice. We report the third case of follicular lymphoma grade 1 limited to the gallbladder , found in the laparoscopic cholecystectomy specimen from a 80 year old woman with symptomatic gallstones. The patient was admitted to the hospital due to the symptoms and signs of acute cholecystitis. Ultrasound examination demonstrated gallstones within the gallbladder, the wall was not thickened. There was no lymph node enlargement in the abdominal cavity. On microscopy, the wall of the gallbladder was extensively infiltrated by lymphoid cells forming neoplastic follicles. The neoplastic infiltrate was composed of small to medium sized centrocytes with cleaved nuclei, inconspicuous nucleoli and scant pale cytoplasm. Additionally, less than to 5 centroblasts per high power field were found throughout the background of cleaved follicle centre-like cells. Immunohistochemical study showed that these cells were positive for B–cell marker CD20, CD79a. The majority of neoplastic cells showed bcl-2 immunoreactivity. Furthermore, they were CD5 and cyclin D1 negative. Our case demonstrates that follicular lymphoma can be limited to the gallbladder and confirm that it can occur in an organ normally devoid of lymphoid tissu. PP3-153 CYCLIN D1 AND TROP-2/GA733-1 IMMUNOHISTOCHEMICAL INVESTIGATION IN ANAPLASTIC LARGE CELL LYMPHOMAS Mattheos Bobos, Vassiliki Kotoula, Vassiliki Kaloutsi, Georgia Karayannopoulou, Ioannis Kostopoulos Department of Pathology, Aristotle University Medical School, Thessaloniki, Greece Background: Anaplastic large cell lymphoma (ALCL) is a rare non-Hodgkin lymphoma with special immunophenotypic and molecular characteristics. The protein Cyclin D1 is a major regulator of the G1 to S phase transition of the cell cycle and a well known proto-oncogene. In lymphomas, up-regulated cyclin D1 protein resulting from the translocation t(11;14)(q13;q32) has been reported in MCL, MM and cases classified as B-CLL and prolymphocytic leukaemia. Recently, has been reported that nuclear expression in 7/10 C-ALCL was detected using polyclonal antibody against cyclin D1. Trop-2/GA733-1, a cell surface glycoprotein, is a calcium signal transducer, may play an important role in the control of cell-cell adhesion and is frequently expressed at high levels by human carcinomas. A fusion transcript between CCND1 and TROP2 genes has been isolated from human tumors and has been found to possess potent transforming activity. Moreover, a cryptic promoter in the coding region of CCND1 gene drives the expression of TROP2 in the absence of exogenous promoters. The aim of the present study was to study the expression of cyclin D1 in ALCL. In addition, we tested for first time in paraffin material the mAb Trop-2 in order to examine the expression profile in ALCL. Methods: We utilized 52 FFPE biopsies classified as ALCLs according to the latest WHO criteria. For the study purposes, whole tissue sections and sections from 41 arrayed ALCL cases were used for IHC. The rabbit monoclonal anti-human cyclin D1 (clone SP4) and the mouse monoclonal anti-human cyclin D1 (clone DCS-6) antibodies were used for detection of cyclin D1 protein. An immunoenzymatic doublestaining method was also performed (Envision G/2 System/AP) for Cyclin D1 (SP4 or DCS-6) and CD30 mAb. For Trop-2/GA733-1 investigation the mouse antihuman mAb (551317, BD) was used. Results: None of the ALCL cases expressed nuclear or cytoplasmic cyclin D1 protein with both mAbs. All the ALCL tested cases were also negative for Trop-2 expression. Trop-2 predominantly membrane staining pattern was observed in squamous cells of the epidermis and in the epithelial cells of merocrine sweet glands. In 2 ALCL cases (1 C-ALCL and 1 systemic ALCL) scattered neoplastic cells showed slight nuclear positivity. Conclusions: The absence of Cyclin D1 protein in our study of ALCL cases is in line with

previous reports. Trop-2 absence in lymphoma cells may indicate that this protein is not involved on the tumor growth. The nuclear staining in rare tumor cells requires further investigation. PP3-154 COMBINED SKIN LESION CONSISTING OF BASAL CELL CARCINOMA AND LANGERHANS CELL HISTIOCYTOSIS Ann Goussia1, Ioannis Nesseris1, Eufemia Balasi1, Efstathios Lykoudis1, Christos Kalogeropoulos2, Maria Bai1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Ophthalmology, Medical School, University of Ioannina, Greece Background. Langerhans cell histiocytosis (LCH) is a neoplastic proliferation of CD1Į, S100-protein positive dendritic Langerhans cells (LCs) characterized by a highly variable clinical presentation and biological behavior. The pathogenesis of the disease is unknown. There is an association of disseminated LCH with other malignancies, mainly leukemias and lymphomas, although many of these cases may be related to therapy. Moreover, uncommonly combined lesions consisting of LCH and lymphomas or solid tumors have been reported in patients without previous history of systemic disease. In these cases the development of systemic LCH has not been described. We report a highly unusual case of combined skin lesion consisting of basal cell carcinoma (BCC) and LCH. Case report. A 63-year-old healthy man presented with a skin nodule involving the upper left eyelid. An excision biopsy was performed. The histopathological findings of the excised lesion (greater diameter:1.4 cm), were those of BCC. However, in the reticular dermis, adjacent to the BCC, a diffuse infiltration composed of large cells with moderate abundant cytoplasm and grooved or indented nuclei was observed. An admixture of small lymphocytes, plasma cells and few eosinophils was also present. Immunohistochemically, the large cells were CD1Į, S-100, CD68, CD3, CD45 and vimentin positive and EMA, pan-CK, MPO, CD30, B and T-cell markers negative. Discussion. Combined skin lesions consisting of LCH and BCC represent an uncommon condition. We are aware of only one similar reported case localized in the scrotum. The nature of LCs in combined lesions remains elusive with debate as to whether they represent a true neoplasm or an exaggerated reactive proliferation, propably induced by factors released by the adjacent tumor cells. X-chromosome inactivation assays in some of these lesions showed that, at least in a proportion of cases, LCs are polyclonal. PP3-155 IMMUNOHISTOCHEMICAL EXPRESSION OF CELL CYCLE REGULATORY PROTEINS IN MULTIPLE MYELOMA Ilias Pessach1, Sevasti Kamina2, Ann Goussia2, Eufemia Balasi2, Evangeli Lampri2, Angelos Skyrlas2, Maria Stoura1, Helen Kapsali1, Panagiotis Kanavaros3, Niki J. Agnantis2, Konstantinos L. Bourantas1, Maria Bai2 1 Department of Hematology, Medical School, University of Ioannina, Greece 2 Department of Pathology, Medical School, University of Ioannina, Greece 3 Department of Anatomy-Histology-Embryology, Medical School, University of Ioannina, Greece Background. The mechanisms involved in cell cycle deregulation in multiple myeloma (MM) are largely unknown. Therefore, the immunohistochemical expression of G1/S cell cycle regulatory proteins were analyzed in relation to the proliferation profile (Ki67, cyclin A, cyclin B1) and clinicopathological parameters in MM. Materials and Method. Bone marrow biopsy specimens from 45 patients with MM were studied by immunohistochemistry, using antibodies directed against cyclin

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D1, cyclin D2, cyclin D3, cyclin A, cyclin B1, Ki67, p53, Rb and p16 proteins. Correlations between continuous variables were calculated using Spearman’s coefficient test and overall survival was estimated using the Kaplan–Meier method. Results. Expression (more than 10% of neoplastic cells) of cyclin D1, cyclin D2, cyclin D3, cyclin A, cyclin B1, Ki67, p53, Rb and p16 proteins was found in 17/45 (37.8%), 18/42(43%), 3/43(7%), 11/44(25%), 0/43(0%), 20/44(45.5%), 17/45(37.8%), 35/44 (79.5%) and 37/44 (84.1%) cases, respectively. The expression of Ki67 protein was positively correlated with the expression of cyclin A and cyclin D2 proteins (p<0.001 and p=0.001, respectively). Moreover, Ki67 and cyclin A expression was increased in parallel with the expression of Rb (p=0.016 and p<0.001, respectively) and p16 proteins (p=0.043 and p=0.005, respectively). A positive correlation was also found between Rb and p16 expression (p=0.003). Although, cyclin D1 and p53 protein were overexpressed, their expression was not correlated with the expression of Ki67, cyclin A or cyclin B1 proteins. The histological stage was positively correlated with Ki67, cyclin A, cyclin D2 and p53 expression (p=0.03, p=0.04, p=0.003 and p=0.026, respectively). Kaplan–Meier analysis showed that the histological stage was significantly associated with patient overall survival (p=0.036). The expression of the studied cell cycle regulatory proteins did not show any prognostic significance. Conclusions. The findings of this study suggest that in MM: a) cyclin D1 and cyclin D2 proteins are overexpressed, b) the expression of cyclin D2 is increased in parallel to proliferative activity and may be involved in the deregulation of cell cycle, and c) the expression of Rb and p16 proteins follows proliferative activity suggesting no abnormal loss of these proteins. PP3-156 MICROVESSEL DENSITY AND EXPRESSION OF THE ANGIOGENIC PROTEINS VEGF, VEGFR1, VEGFR2 AND HIF-1Į IN HODGKIN AND REED-STERNBERG CELLS OF CLASSICAL HODGKIN LYMPHOMA Evangeli Lampri1, Nafsika Simou1, Evangelos Briasoulis2, Ann Goussia1, Sevasti Kamina1, Lazaros Nikiforidis2, Asimina Demou3, Vasiliki Malamou-Mitsi1, Nikolaos Pavlidis2, Niki J. Agnantis1, Panagiotis Kanavaros4, Maria Bai1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Oncology, Medical School, University of Ioannina, Greece 3 Department of Pathology, Ioannina General Hospital G. Hatzikosta, Greece 4 Department of Anatomy-Histology-Embryology, Medical School, University of Ioannina, Greece Background. Few studies have focused on angiogenesis in classical Hodgkin lymphoma (cHL). Therefore, in the present study the microvessel density (MVD) and the immunohistochemical expression of angiogenesis relatedproteins in Hodgkin and Reed-Sternberg cells (HRS) of cHL were analysed. Materials and Method. Biopsy material from 95 cases with cHL (mixed cellularity:60, nodular sclerosis:23, lymphocyte depletion:3, unclassified:9) were studied immunohistochemically for the expression of VEGF, VEGFR1, VEGFR2 and HIF1a proteins. In addition, MVD was estimated after immunostaining for CD34 (MVD-CD34) and CD105 (MVD-CD105) proteins and evaluated as the mean (±SD) vessel count/high-power microscopic field (HPF). The results were correlated with clinicopathological data. Results. Expression (at least 10% positive HRS cells) of VEGF, VEGFR1, VEGFR2 and HIF1a proteins was found in 78/90 (86.7%), 32/95 (33.7%), 18/89 (20.2%) and 46/81 (56.8%) cases, respectively. The MVDCD34 and the MVD-CD105 were 61(±28.7) and 12(±7.65), respectively. VEGF expression was positively correlated with VEGFR2 expression and MVD-CD34 (p=0.003 and p=0.02, respectively). A similar correlation was found between VEGFR1 expression and MVD-CD34 (p=0.04). Moreover, an inverse

correlation between the expression of VEGFR2 protein and the clinical stage of the disease was found (p=0.02, r=-0.505). Conclusion. The present study shows that the VEGF, VEGFR1, VEGFR2 and HIF1a proteins are expressed in HRS cells of cHL. VEGF produced by HRS cells may act in a paracrine and autocrine fashion in cHL and possibly is implicated in the pathogenesis of the disease. PP3-157 ALTERATIONS OF THE p53, Rb AND p27 TUMOR SUPPRESSOR PATHWAYS IN DIFFUSE LARGE B-CELL LYMPHOMAS Elena Tsanou1, Ann Goussia1, Angelos Skyrlas1, Sevasti Kamina1, Ioannis Sainis2, Eufemia Balasi1, Niki J. Agnantis1, Panagiotis Kanavaros1, Maria Bai1 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Anatomy-Histology-Embryology, Medical School, University of Ioannina, Greece Background. Diffuse large B-cell lymphomas (DLBCL) display defects in cell cycle and apoptosis regulation. Therefore, the immunohistochemical expression patterns of the proteins p14, p21, Hdm2 and cyclin D2 were analyzed in relation to the previously reported expression of other major cell cycle proteins (p53, Rb, p16, p27, Ki67 and cyclins A, B1, D2, D3 and E), apoptosis-associated proteins (bcl2, bcl-xl, bax, bak, bad and bid) and the B-cell differentiation immunophenotypes. Materials and Method. Seventy nine cases of de novo DLBCL (37 nodal and 42 extranodal) classified according to WHO classification were studied by immnohistochemistry, using antibodies directed against p14, p21, Hdm2 and cyclin D2 proteins. Results and Conclusions. Expression of the proteins p14, p21, Hdm2 and cyclin D2 was observed in 62/71 (87%), 22/76 (29%), 35/74 (47%) and 11/77 (14%) cases, respectively. Immunohistochemical alterations of the p53 [p53-Hdm2-p21p14], Rb [Rb-p16-cyclin D (D2 or D3)] and p27 [p27-cyclin E] pathways were found in 56/77 (73%), 53/79 (67%), 54/79 (68%) cases, respectively. Concomitant alterations of the p53-Rb, p53p27 and Rb-p27 pathways were found in 40/77 (52%), 38/77 (50%) and 36/79 (46%) cases, respectively. Three concomitant alterations of p53-Rb-p27 pathways were found in 28/79 (35%) cases. The main findings of the present study were the following: 1) alterations of the p27 pathway were associated with higher expression of Ki67 (p=0.023), 2) concomitant alterations of the p53-Rb pathways and the p53-p27 pathways were associated with higher expression of cyclin A (p=0.015 and p=0.021, respectively) and 3) concomitant alterations of the p53, Rb and p27 pathways were associated with higher expression of cyclin A (p=0.013). Since cyclin A supports DNA replication, centrosome duplication and mitosis, these findings indicate that concomitant alterations of the p53, Rb and p27 pathways in DLBCL may have cooperative effects resulting in increased neoplastic cell proliferation. This might explain, at least partially, the association between concurrent aberrations of the p53, Rb and p27 pathways and aggressive clinical behavior in DLBCL. PP3-158 ANGIOGENESIS AND MYELOMA : A COMPARATIVE STUDY OF CD34 AND CD138 IN PRE- AND POSTTREATMENT BONE MARROW BIOPIES Karima Mrad1, Maha Driss1, Salwa Ladeb2, Neziha Ben Hamida1, Houda Arbi1, Radhia Ben Ghorbel1, Houda Azaiez1, Mongia Zitouni1, Abdelrahman Abdelkefi2, Khaled Ben Romdhane1 1 Salah Azaiez institute, Tunisia 2 Centre de Greffe de Moelle Osseuse, Tunisia Background and aims: Bone marrow (BM) angiogenesis is increased in multiple myeloma and is an important prognostic factor for survival. Given its potential antiangiogenic effect, we

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evaluated if thalidomide therapy would affect the BM microvessel density (MVD). Material and methods : We tested both CD34 and CD138 on paraffin sections of bone marrow biopsy using a labelled streptavidin-biotin peroxydase method. MVD and % of CD138 positive cells were compared between pretreatment biopsies and those obtained following therapy. Slides stained with antibodies to CD34 were scanned under low power (x100) to identify three hot spots or areas with the greatest number of microvessels. These three areas were then evaluated at x400 magnification. Large vessels and vessels within the bony spicules and under the periosteum were excluded. Areas of staining without discrete breaks were counted as single vessels. The presence of a lumen or red cells was not required for the identification of microvessels; any highlighted endothelial cell or cell cluster separate from the adjacent microvessels was counted as a distinct vessel. The number of vessels in one x400 field was determined for each hot spot; the average was taken as the MVD. Results : On the basis of the MVD, the patients were grouped into high- and low grades using the mean MVD as the cutoff. The mean MVD for the entire group was 18 (range, 3–54). Good responders (9 patients) with a mean decrease of 60% in CD138 cells (range 30-100%) had a low-grade MVD (mean 15) and a decreasing MVD (pre-treatment MVD : 19, post-treatment MVD : 10). Patients with disease progression (5 patients) had a high MVD (mean 23) with an increasing MVD (pre-treatment MVD : 20, post-treatment MVD : 26). Conclusion : Our results confirm a significant decrease of MVD with thalidomide therapy. Although not conclusive, this result lends further support to the hypothesis that angiogenesis is a relevant therapeutic target in myeloma. PP3-159 MYELOID SARCOMA OF THE LYMPH NODE WITH EXTENSIVE NECROSIS Hulya Ozturk Nazlioglu1, Tulay Ozcelik2 1 Uludag University Medical School, Department of Pathology, Bursa, Turkey 2 Uludag University Medical School, Department of Hematology, Bursa, Turkey Myeloid sarcoma is a tumour mass of myeloblasts or immature myeloid cells occurring in an extramedullary site. The tumour mass may precede or occur concurrently with acute or chronic myeloid leukemia. A 38-year-old woman was admitted to hospital because of a left axillary lymph node enlargement. She was previously diagnosed as having Acute Myeloid LeukemiaM5 (AML-M5) and was treated with chemotherapy. Fine-needle aspiration cytology of the lymph node was consistent with lymphoma. Excisional biopsy specimen of the lymph node revealed diffuse infiltration of myeloblasts positive for CD43 and myeloperoxidase staining on immunohistochemistry, and changed the diagnosis from malignant lymphoma to myeloid sarcoma. We report an unusual case of acute myeloid leukemia presenting with myeloid sarcoma with a lymph node lesion exhibiting extensive lymph node infarction. A high index of suspicion for myeloid sarcoma is necessary to avoid missing this lesion. The critical factors for detecting this tumor are to be aware of this disease, cooperation between clinician and pathologist and the application of spesific immunohistochemical antibodies to detect the myeloid origin. PP3-160 EPSTEIN-BARR VIRUS AND HUMANHERPES 8 IN LYMPHOID NEOPLASMS FROM SPAIN Ramon Bosch1, Marylene Lejeune1, Lluis Pons1, Eva Domingo2, Silvia De Sanjose3, Yolanda Benavente3, Vicens Romagosa4, Juan Jose Sirvent5, Monica Garcia-Cosio6, Vanessa Morente7, Joaquin Jaen1, Xavier Ortin8, Barbara Tomas1, Mª Teresa Salvado1, Tomas Alvaro1 1 Pathology Department Hospital de Tortosa Verge de la Cinta, Spain 2 Hematology Department CSUB Barcelona, Spain

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Catalan Institute of Oncology, Barcelona, Spain Pathology Department CSUB Barcelona, Spain 5 Pathology Department Hospital Joan XXIII de Tarragona, Spain. 6 Pathology Department Hospital Ramon y Caja Madrid, Spain 7 Pathology Department Hosptial Sant Joan de Reus, Spain 8 Hematology Department Hospital de Tortosa Verge de la Cinta, Spain 4

INTRODUCTION: Epstein-Barr virus (EBV) and Human Herpesvirus 8 (HHV8) are well known lymphotropic oncogenic agents. Our goal was to identify the proportion and the histological types of lymphoid neoplasms that express EBV and HHV8 in tumoral cells of a large series of cases diagnosed in Spain. MATERIAL AND METHOD: Tissue microarrays (TMA) were constructed from paraffined tumoral tissue of lymphoid neoplasms diagnosed in 5 Spanish hospitals from 1998 to 2003. EBER in situ hybridization for EBV and LANA immunohistochemistry for HHV8 were performed on histological sections of the TMA. RESULTS: From 182 obtained biopsies, 24% were Hodgkin’s lymphomas (HL) and 76% were nonHodgkin’s lymphomas (NHL). 93,5% of NHL were of B cell origin whereas 6,5% were of T/NK cell. HLs were all HHV8 negative while only one case of NHL was HHV8 positive (an extracavitary PEL in a HIV+ patient). 22,7% of HL and 10,9% of NHL were EBV positive (EBV+). 19,4% (13/67) of the highgrade B-cell NHLs (Diffuse large B-cell, Burkitt and Burkitt-like) were EBV+ while all the low-grade B-cell NHL (0/60) were EBV negative. 11,1% (1/9) of T/NK NHL were EBV+. 10% of the cases were associated with an immunodeficiency and showed an expression significantly higher of EBV than cases from immunocompetents (9.1% vs. 55,6% p<0.0001). In the immunocompetent group, there was a significant difference between HLs and NHLs EBV expression (21,4% vs. 4.9%; p=0.003) and, by histological subtypes, the high-grade B-cell NHLs were EBV+ in a 9,3% of the cases and, in this group, diffuse large B-cell NHL showed an expression of 7,7% (4/52). CONCLUSIONS: When studied with TMA, the expression of HHV8 in the tumoral cells of lymphoid neoplasms is very infrequent (0.54% of lymphoid neoplasms) and when found, an underlying immunodeficiency should be excluded. By contrast, EBV is expressed in a higher proportions, mostly in HLs and high-grade B-cell NHLs and, in a significantly higher proportion, in the setting of an immunodeficiency. (PI 04-0091, PI 04/1440, 04/1467, 05/1527) PP3-161 MACROPHAGES AND DENDRITIC CELLS IN THE MICROENVIROMENT OF THYMIC NEOPLASMS. CORRELATION TO VASCULAR DENSITY AND HISTOLOGIC TYPE Vassiliki Tzelepi1, Vassiliki Zolota1, Athanassios Tsamandas1, Paraskevi Zyli1, Georgia Sotiropoulou Bonikou2 1 Department of Pathology, Medical School, University of Patras, Greece 2 Department of Anatomy, Medical School, University of Patras, Greece Background: Tumor associated macrophages (TAMs) are key components of the tumor microenvironment and seem to promote tumor growth. Their number is increased in various neoplasms. Dendritic cells (DCs) are basic components of the innate inflammatory response. Their role in tumor microenviroment is unclear. The presence of TAMs and DCs in thymic neoplasms has not been thoroughly evaluated before. This study investigates the presence of TAMs and DCs in thymomas and correlates them with microvessel density (MD). Method: A total of 20 thymomas (4AB, 2B1, 4B2, 6B3 and 4C according to WHO histologic classification and 3T1, 10T2, 4T3 and 3T4 according to TNM staging) were immunostained using antibodies to S100, PGM1, CD34 and CD31 proteins. For all antibodies, areas with the

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highest density of positive cells were selected at low power magnification (x40), and the total number of immunostained cells or vessels was counted at high power (X400). At least three high power fields were counted and the mean value was marked as labeling index (LI). Results: LIs for microvessels according to histologic subtype were as follows (mean value±SD): AB 73.25±23.78, B1 54±43.8, B2 42.2±26.9, B3 32.83±24.02 and C 36.5±24.26. Tumors were subdivided for statistical purposes in two groups. Surprisingly, AB, B1 and B2 thymomas demonstrated higher MD compared to B3 and C (58.9±28.75 vs 34.3±22.8, respectively, p=0.049). S100 positive DCs were always identified within the tumor parenchyma. DCs number was higher in AB, B1 and B3 (17.125±12.26) compared to B3 and C (3.3±2.62) thymomas (p=0.003). A positive correlation between DCs and MD was also recorded (p=0.007). PGM1 positive TAMs were diffusely found in the tumor parenchyma in all AB, B1 and B2 tumors. In contrast, 50% of the B3 and C thymomas exhibited a tumor-stroma interface preference for TAMs localization. No statistical correlation between histologic type and TAMs number was seen [AB/B1/B2 45.67±16.67, B3/C 39.10±18.6336 (p=0.432)]. In addition, MD, DCs and TAMs were not correlated to tumor stage. Conclusions: A decrease of DCs is noted in less differentiated thymomas and implies that impairment of the host defense accompanies and possibly supports the progression of thymic neoplasia. TAMs do not seem to play an important role in thymic tumors, albeit their presence at the invasion front in the less differentiated tumors suggests a possible role in tumor invasion. Finally, further studies are needed to elucidate the exact role of vascular density in thymomas. PP3-162 IMMUNOHISTOCHEMICAL EXPRESSION PATTERNS OF NEURAL AND NEUROENDOCRINE MARKERS, NEURAL GROWTH FACTOR RECEPTORS AND THE TUBULIN-ȕ2 AND -ȕ4 ISOTYPES IN THYMUS Michael Doukas1, Alexandra Papoudou-Bai1, Georgios Karatzias2, Ann Goussia1, Kalliopi Stefanaki3, Yiotanna Dalavanga2, Niki J. Agnantis1, Maria Bai1, Panagiotis Kanavaros2 1 Department of Pathology, Medical School, University of Ioannina, Greece 2 Department of Anatomy-Histology-Embryology, Medical School, University of Ioannina, Greece 3 Department of Pathology, Agia Sophia Hospital of Athens, Greece Background. Increasing evidence suggest that neuroimmune networks play key roles in the thymic histophysiology and pathology. Therefore, we analyzed the distribution of human thymic cells expressing major neural and neuroendocrine markers and the neural growth factor receptors (NGFR) in combination with the expression patterns of various cytokeratins. Additionally, since some tubulin-ȕ isotypes are preferentially expressed in neuronal cells, we analyzed the immunotopographical distribution of thymic cells expressing tubulin-ȕ2, -ȕ3 and -ȕ4. Materials and Method. Twenty cases of normal thymuses were analyzed by immunohistochemistry for the expression of the protein gene product 9.5 (PGP 9.5), chromogranin A (CHRA), synaptophysin (SYN), neuron specific enolase (NSE), tyrosine hydroxylase (TH), CD56, CD57, neurofilaments (NFs) (140160kDa), the NGFRs (TrKA and p75), the tubulin-ȕ2 and -ȕ4 isotypes and the cytokeratins (CKs) 7, 8, 10, 13, 14, 18 and 19 proteins. Results. Thymic epithelial cells (TEC) expressed PGP 9.5, CHRA, SYN, NSE, TH, CD56, CD57, NFs, TrKA, p75, tubulin-ȕ2 and -ȕ4 isotypes and the CKs 7, 8, 10, 13, 14, 18 and 19. PGP 9.5 was preferentially expressed in cortical TEC whereas SYN, CHRA, NSE, TH and NFs were preferentially expressed in medullary TEC and Hassall corpuscles. Variable levels of expression of tubulin-ȕ2 and -ȕ4 were observed in all TEC subtypes whereas tubulin-ȕ3 was undetectable in TEC.

Subcapsular and cortical TEC display higher expression levels of tubulin-ȕ4 and lower expression levels of tubulin-ȕ2 in comparison to those observed in medullary TEC and Hassall corpuscles. Conclusions.The diversity of the immunotopographical distribution and the expression levels of neural and neuroendocrine markers, the NGFRs, the tubulin-ȕ2 and -ȕ4 isotypes and the CKs in the distinct subtypes of TEC may reflect the diversity of their biological functions and/or their different stages of differentiation. The present results provide further immunohistological evidence that numerous neural and neuroendocrine factors may be required for the development and the function of the human thymic microenvironment. PP3-163 CIRCULATING LEVELS AND PROGNOSTIC SIGNIFICANCE OF VASCULAR ENDOTHELIAL GROWTH FACTOR AND ITS RECEPTOR IN CHILDHOOD ACUTE LEUKEMIA Mitra Mehrazma, Khadije Arjomandi, Behzad Torkamani Poor Iran University of Medical Sciences, Ali-Asghar Hospital, Iran BACKGROUND:Vascular endothelial growth factor (VEGF) and its receptors (VEGF-R1 and R2) are major regulators of angiogenesis, which plays a key role in the growth and dissemination of solid tumors and hematologic neoplasms. METHOD: To assess serum VEGF and VEGF-R1 levels and their prognostic significance in newly diagnosed acute leukemias (ALL and AML), we used a radioimmunoassay (RIA) to quantify their levels in stored samples obtained before treatment from 53 pediatric patients. Bone marrow blast percentage was counted at day of diagnosis and 2 weeks after induction therapy . RESULT:s VEGF level was between 2.7 and 138 pg/ml sVEGF-R1 level was between 0.0020 and 0.1836 ng/ml. A reverse correlation was observed between plasma sVEGF and especially sVEGFR1 levels and the rate of complete remission in pediatric patients with acute leukemias. CONCLUSION: Concentration of sVEGF and sVEGF-R1 in plasma should be considered a significant factor in response to treatment and considerable reduction of bone marrow blast percentage. In contrast, there was no relationship between their levels and WBC or blast count at diagnosis, or between VEGF level and such established prognostic factors as age, cytogenetics or FAB category. PP3-164 PRIMARY LYMPHOMA OF THE THYROID GLAND. A CLINICAL PATHOLOGICAL REVIEW OF 3 CASES SEEN AT THE PATHOLOGY UNIT OF THE GENERAL HOSPITAL OF MEXICO Monica Romero1, Dulce Maria Macias1, Marco Antonio Duran2, Mercedes Herrnandez1 1 Hospital General de Mexico, Mexico 2 Hospital General de Mexico, Mexico Introduction Primary lymphomas of the thyroid gland constitute approximately 8% of all thyroid neoplasms. They are often associated (80% of the time) to Hashimoto and lymphocytic thyroiditis. The most frequent histological types are: large cell diffuse lymphoma B, marginal zone MALT type lymphoma, follicular lymphoma, plasmacytoma and peripheral t/nk lymphoma. Material and methods A retrospective review (20002005) was done of all pathological surgical sample records of the thyroid gland. A total of 115 cases of thyroid neoplasms were found: 64 (5.7%) corresponded to thyroiditis, 26 (40%) to lymphocytic thyroiditis, 24 (37.5%) to Hashimoto´s thyroiditis and 14 (21.8%) to unspecified chronic thyroiditis. Three lymphomas were found which had been submitted to the following specific immunohistochemical tests: cd 20, cd79a, cd 45, cd45 RO, cd2, bcl2, bcl6. cd 5, VEB, calcitonin, chromogranin and specific neuron enolase. Results The three lymphoma patients were females 56, 58 and 70 years of age. The symptomatology constituted a gradual and symmetrical growth of

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the thyroid gland, with one of the patients having had cervical compression syndrome. Two cases corresponded to large cell diffuse lymphoma of the centroblastic variety. No marginal zone lymphoma cases nor thyroiditis were found, while one Malt type of the marginal zone associated to Hashimoto thyroiditis having LBDCG transformation zones was found. All were positive for cd 20 and two for bcl2 and negative for the rest of the antibodies. Conclusions Primary thyroid lymphomas were found in 2% of the cases we reviewed and are considered infrequent. The associated diseases, lymphoid and Hashimoto thyroiditis were also found to be infrequent (5.7%). This was probably due to the low frequency of primary thyroid lymphomas in our series. PP3-165 IMMUNOEXPRESSION OF Skp2 AND CUl1, MOLECULES IMPLICATED IN p27Kip1 DEGRADATION IN AGGRESSIVE AND INDOLENT B-CELL LYMPHOMAS Chaido Sirinian1, Nikolaos Giannakoulas2, Panagiotis M. Zikos3, Panagiota Matsouka2, Maria Melachrinou1 1 Department of Pathology, University of Patras Medical School, Patras, Greece 2 Department of Internal Medicine, Division of Hematology, University of Patras Medical School, Patras, Greece 3 Hematology Division, “Agios Andreas” General Hospital of Patras, Patras, Greece Background: Reduced levels of p27Kip1 (p27) are frequently observed in human cancers and associated with aggressive tumor behavior and poor prognosis. There is evidence that Skp2, a member of ubiquitin-ligase SCF (Skp1-Cul1-Rbx1-F box) complex, promotes p27 degradation. Moreover, Skp2 deregulation has been implicated in oncogenesis and lymphomagenesis. The aim of this study was to investigate the expression of Skp2 and Cul1, components of ubiquitin-ligase SCF complex, in aggressive and indolent B-cell lymphomas, and compared them with p27 expression and proliferation index (PI). In addition, the prognostic significance of Skp2 and/or p27 expression was studied in cases with available clinical data. P21Cip1 (p21) expression was also examined, because p21 is a good substrate for the ubiquitin-ligase SCF complex. Method: Formalin-fixed, paraffin-embedded tissue from 126 B-cell lymphomas [64 aggressive (DLBCLs) and 62 indolent (18 SLLs, 14 FLs, 11 MCLs, 19 MZLs)] were immunostained with antibodies for Skp2, Cul1, p27, p21 and MIB-1 using Envision detection kit. Clinical data for statistical analysis were available for 87 cases (40 aggressive, 47 indolent). The median follow up period was 33 months (22 mo for aggressive, 40 mo for indolent). Results: The mean value of Skp2 immunoexpression in aggressive and indolent lymphomas was 33.2% and 7.7%, respectively (p<0.001). The corresponding values for Cul1 were 36.4% and 14.7% (p<0.001), for p27 32.5% and 62.6% (p<0.0001), for p21 13.7% and 6.6% (p=0.025) and for MIB-1 68.7% and 27.1% (p<0.001). Skp2 showed an inverted correlation with p27 (p=0.035) and a positive correlation with Cul1 (p=0.002) and PI (p=0.007) in aggressive lymphomas. In addition, Skp2 had a similar correlation with p27 (p=0.025) and PI (p<0.001), but not with Cul1, in indolent lymphomas. There was also an inverted correlation between the expression of p27 and PI (p=0.008). Overall probability of survival was 35% for the whole group. This was 51.7% and 31.5% for patients with aggressive and indolent lymphomas, respectively (p=0.074). Skp2 and p27 expression did not correlate patients’ prognosis. Conclusion: Our findings demonstrate a significant positive correlation between Skp2 expression and proliferation index and suggest the p27 degradative function of Skp2 in aggressive and indolent lymphomas. The positive relationship between Skp2 and Cul1 expression in aggressive lymphomas indicates their implication in p27 degradation.

PP3-166 PRIMARY BREAST LYMPHOMAS. A FOUR CASE REPORT Monica Romero, Mercedes Hernandez, Marco Antonio Duran, Avissai Alcantara Hospital General de Mexico, Fac. de Medicina, Mexico Introduction The lymphomas that affect breasts may be primary or infiltrates as systemic manifestations of the disease. The most common histological types are large cell diffuse B lymphomas and extra-nodal B mucosa-associated lymphatic tissue lymphomas (marginal zone or extra-nodal lymphomas). Material and methods This is a report of four cases with primary breast lymphomas. The criteria for diagnosis were based on Wiseman´s and Liao´s proposal which includes: 1) specimen biopsy that involves a lymphoma in close relation to breast parenchyma and 2) no evidence of systemic disease at diagnosis. The clinical and histological data were reviewed, as well as immunohistochemical reactions. Results The average age of the patients was 29 years old. Three patients had problems with their right breasts and another bilaterally. There was progressive increase in volume, pain at palpation and skin affection, in addition to purulent secretion from the nipple. No affected lymph ganglia were palpated in three of the patients. Three of the patients died a few months (2-5) after diagnosis with infiltrations to their central nervous system. All four were large cell diffuse lymphomas of immunophenotype B, three were centroblastic, one had clear cells and another had extensive areas of necrosis with a starry-skied focal pattern simulating Burkitt´s lymphoma. They were positive to cd 20, cd 79 a, bcl2, bcl6, and cd 10. Conclusions Primary lymphomas of the breasts are infrequent. In our cases, the lymphomas were seen in young women. One was associated to pregnancy and another to AIDS. The differential diagnosis should be done with scarcely differentiated lobule and duct carcinoma for which immunohistochemistry reactions are very useful. PP3-167 LANGERHANS CELL SARCOMA EXPRESSING CD56 IN A 60 YEAR OLD PATIENT WITH MYELODYSPLASTIC SYNDROME Konstantinos Giaslakiotis1, Athina Androulaki1, Theodora Papadaki2, Pinelope Pinelope Korkolopoulou3, Eustratios Patsouris3 1 Department of Pathology, Laiko University Hospital, Athens, Greece 2 Hemopathology Department, Evangelismos Hospital, Athens, Greece 3 Department of Pathology, Medical School, National and Kapodistrian University of Athens, Athens, Greece BACKGROUND: Langerhans cell tumors as currently defined in the WHO classification are a rare group of neoplastic proliferations showing CD1a and S-100 protein expression and displaying Birbeck granules by ultrastructural examination. Langerhans cell sarcoma (LCS) is the aggressive counterpart showing overtly malignant cytologic features and increased mitotic activity. The association of myeodyplastic syndrome with Langerhans cell tumors is extremely rare. Recently it has been suggested that CD56 expression is related to aggressive biological behavior in Langerhans cell tumors. We report a case of LCS expressing CD56 in a 60 year old patient with myelodysplastic syndrome initially diagnosed in the lymph node diagnostic material. METHODS: Formalin fixed, paraffinembedded tissue sections were stained with H&E and immunohistochemically according to established protocols. RESULTS: Histological examination revealed effacement of the normal structure by a neoplastic proliferation showing nodular and sinusoidal growth pattern. Tumor cells ranged between round and oval displaying a lobulated nuclei with delicate nuclear membrane, chromatin abnormalities and prominent nucleoli. Distinct features of Langerhans cell tumors with the characteristic

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nuclear grooving also were encountered, although infrequently. Eosinophils were rarely scattered. The mitotic rate was 10/10 HPF. Immunohistochemically, tumor cells were strongly positive for CD1a and S-100 protein and slightly positive for vimentin. CD56 was strongly positive. Only very weak CD68+ cells were seen. The proliferation marker Ki67 (MIB-1) reached 15%. Tumor cells failed to express CD3, CD20, CD21, CD35, CD30, CD34, pancytokeratin, HMB-45, Melan A and chromogranin. CONCLUSION: Our report further supports the association of Langerhans cell tumors with myelodyplastic syndrome, reinforcing the hypothesis of a common stem cell disorder. CD56 expression could enhance the prediction of the potential of an aggressive biological behavior in Langerhans cell tumors with borderline morphological features of malignancy. PP3-168 EXPRESSION AND PROGNOSTIC SIGNIFICANCE OF SURVIVIN AND P 53 IN HODGKIN LYMPHOMAS Suheyla Bozkurt1, Nagehan Barisik2, Isik Kaygusuz3, Mahmut Gumus4, Emine Bas1, Sezer Gezer2, Mahmut Bayik3, Tulay Tecimer1 1 Marmara University Pathology Department, Turkey 2 Dr. Lutfi Kirdar Kartal Research and Training Hospital Pathology Department, Turkey 3 Marmara University Hematology Department, Turkey 4 Dr. Lutfi Kirdar Kartal Research and Training Hospital Oncology Department, Istanbul, Turkey Aim: Apoptosis represents a critical pathway whose abnormalities are linked with carcinogenesis. p 53, tumor supressor gene and inducer of apoptosis, suvivin a member of the inhibitors of apoptosis protein family are important regulators whose abnormal expression or mutation is correlated with several human cancers. To investigate their relationships to carcinogenesis and/or prognosis of Hodgkin lymphomas we analyzed their immunohistochemical expression in Hodgkin lymphomas. Material-Method: Paraffin sections from 56 Hodgkin lymphomas (28 noduler sclerosis, 18 mixed cellularity, 4 lymphocyte-rich, 1 nodular lymphocyte predominant, 5 classicalnot specified) were examined with monoclonal antibodies antip53 and survivin using streptavidin-biotin immunohistochemical tecnique. Survivin was evaluated for both staining density and percentage of staining. Staining density evaluated as fallows:negative, mild, moderate, severe; percentage evaluates as fallows: +,<25%; ++, 25-50%; +++, 50-75%; ++++, >75% of Hodgkin/Reed Sternberg cells positive. p 53 was evaluated as fallows: negative <10%; +,10-25%; ++, 25-50%, +++, 5075%;++++ ,>75% of Hodgkin/Reed Sternberg cells positive. Results: p 53 protein was detected in 20 cases. Survivin expression was detected in all cases with variable values. Survivin expression showed significant positive correlation with only age of the patients(p<0.05). Expression of survivin and p53 did not showed significant correlation with subtypes, stage, survival time and recurrences in Hodgkin lymphomas(p>0.05). Conclusion: Altough survivin and p53 expression was detected in Hodgkin lymphomas, these results showed that these markers not considered as a prognostic factors for Hodgkin lymphomas. PP3-169 EXPRESSION AND PROGNOSTIC SIGNIFICANCE OF COX-2 AND P 53 IN HODGKIN LYMPHOMAS Nagehan Barisik1, Suheyla Bozkurt2, Mahmut Gumus3, Isik Kaygusuz4, Nimet Karadayi1, Emine Bas2, Mahmut Bayik4, Tulay Tecimer2 1 Dr. Lutfi Kirdar Kartal Research and Trainig Hospital Pathology Department, Turkey 2 Marmara University Pathology Department, Turkey 3 Dr. Lutfi Kirdar Kartal Research and Trainig Hospital Oncology Department, Turkey 4 Marmara University Hematology Department, Istanbul, Turkey

Aim: Cyclooxigenase(COX) is a rate-limiting enzyme that plays an important role in the inflammatory process and catayses the conversion of arachidonic acid to the prostaglandins and tromboxanes. One of two isotopes, COX-2 is usually not detectable in normal tissues. COX-2 is associated various human cancers especially colorectal adenocarcinomas and it is related to tumor progression. p 53, tumor supressor gene and inducer of apoptosis whose abnormal expression is correlated with several human cancers. To investigate their relationships to carcinogenesis and/or prognosis Hodgkin lymphomas we analyzed their immunohistochemical expression in Hodgkin lymphomas. Material-Method: Paraffin sections from 40 Hodgkin lymphomas (20 noduler sclerosis, 14 mixed cellularity, 2 lymphocyte-rich, 4 classical-not specified) were examined with monoclonal antibodies anti-p53 and COX-2 using streptavidinbiotin immunohistochemical tecnique. COX-2 was evaluated for both staining density and percentage of staining. Staining density evaluated as fallows:negative, mild, moderate, severe; percentage evaluates as fallows: +,<25%; ++, 25-50%; +++, 50-75%; ++++, >75% of Hodgkin/Reed Sternberg cells positive. p 53 was evaluated as fallows: negative <10%; +,10-25%; ++, 25-50%; +++, 50-75%;++++ ,>75% of Hodgkin/Reed Sternberg cells positive. Results: p 53 protein was detected in 14 cases. COX-2 expression was detected in all cases except one with variable values. Expression of COX-2 and p53 did not showed significant correlation with subtypes, stage, survival time and recurrences in Hodgkin lymphomas(p>0.05). Although it was not statisticaly significant, COX-2 expression was higher in nodular sclerosis and mixed cellularity subtypes compared to lymphocyte rich and classical-not specified subtypes(p=0.054). Conclusion: The expression of COX-2 and p53 is confirmed in H&RS cells of classical Hodgkin lymphoma. But, due to absence of association with stage, survival and recurrences of the disease these markers not considered as a prognostic factors for Hodgkin lymphomas. PP3-170 HISTOLOGICAL ASPECTS OF FOLLICULAR DENDRITIC CELL SARCOMA AND PITFALLS IN ITS DIAGNOSIS Petar Cvetkovski1, Ljube Ivkovski2, Vladimir Popovski3, Oliver Karanfilski4, Gordana Petrusevska1 1 Institute of Pathology, Faculty of Medicine, Skopje, Macedonia 2 Laboratory for Histopathology, Institute of Oncology, Skopje, Macedonia 3 Clinic for Maxillofacial Surgery, Skopje, Macedonia 4 Clinic for Haemathology, Clinical Center, Skopje, Macedonia Follicular dendritic cell sarcoma (FDCS) is a neoplastic proliferation of spindle to ovoid cells with morphologic and phenotypic features of follicular dendritic cells. It is a rare neoplasm with most studies representing single case reports or small series. The aim of this presentation is to present a rare case of FDCS in 27 year-old woman, previously diagnosed and treated as B cell immunoblastic non-Hodgkin`s lymphoma. The first diagnosis was made on a cervical lymph node biopsy five years ago, when she was treated with five cycles of CHOP regimen and complete remission of the disease followed. Four years later new tumor mass in the left midneck region appeared and was surgically treated. On the dissection one could barely recognize destroyed architecture of lymph node by infiltrative fleshy ten grey tissue that even involve subcutaneous and dermal tissue. There was no ulcerative lesion. Histological examination of the tissue sections stained with HE, Giemsa, Reticulin, PAS and Van Gieson revealed spindle to ovoid cell proliferation arranged in fascicles and storiform pattern. There were areas with plexiform architecture and myxomatous degenerative changes. In some of the cells there were PAS negative clear vacuoles. The cells had elongated oval nuclei with granular chromatin and small but distinct nucleoli enclosed in lentiform and syntitial slightly eosinophylic cytoplasm. There were fluid-filled cystic spaces with angiomatous features and only small solid areas with large

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pleomorphic cells were found. Mitotic index was low. These tumor cells enclosed fibrous trabeculae with blood vessels surrounded by cuff of lymphocytes. Epidermal layer of the skin was intact but there was infiltration in the lower papillary dermis. Immunohistochemical analysis revealed positivity of the cells for Vimentin, HLA DR, CD21, CEA (nonspecific?) and spotty positivity for ESA. The stainings for cytokerains, S100, HMB45 and lymphocytic subpopulations were negative. Ki67 evaluated low proliferative index of 15%. Based on HLA DR and CD21 expression and the morphological appearance we made a diagnosis of FDCS and revised the previous diagnosis of B immunoblastic lymphoma. We found the same immunophenotype of the neoplastic cells that morphologically resemble large immunoblastic cells. However they were CD20 negative and CD21 positive. We can conclude that FDCS could have pleomorphic expression that ranges from lymphoid like appearance to spindle cell like or neurogenic-like neoplastic lesion. Immunohistochemical analysis is very helpful in distinguishing various entities. PP3-171 HUMAN IMMUNODEFICIENCY VIRUS-ASSOCIATED LYMPHOMAS. RECLASSIFICATION WITH TMABASED IMMUNOHISTOCHEMISTRY AND MOLECULAR BIOLOGY, ITS ASSOCIATION TO EPSTEIN-BARR VIRUS INFECTION AND EXPRESSION OF NUCLEAR FACTOR KAPPA-B AS PROGNOSTIC FACTORS Daniel Montante Montes De Oca, Fredy Chablé Montero, César Lara Torres, Braulio Martinez Benitez, Nancy Camorlinga Tagle, Brenda Marquina Morán, Arturo Ángeles Ángeles, Carmen Lome Maldonado Department of Pathology. Instituto Nacional de Ciencias Medicas y Nutricion, Mexico Objectives: To know the clinical and pathological features of human immunodeficiency virus (HIV)-associated lymphomas, its association to Epstein-Barr virus (EBV) infection and expression of nuclear factor kappa-B (NF-kB) as prognostics factors. Material and Methods: HIV-associated lymphomas were selected in a period of 18 years. All the cases were reviewed, and they were reclassificated on the basis of morphology, TMA-based immunohistochemistry and molecular biology. Clinical features were obtained from the clinical records. Results: Fifty-seven cases were obtained. The average age of presentation was 40 (+/10) years, 95% were men and 5% women; 92.9% cases were in C3 stage. Seventy-eight patients presented B symptoms. The length of symptoms was from 0 to 48 months, with average of 6 months. The most frequent anatomic localization was nodal in 22 cases, oral cavity, CNS, colon, and rectum in 6 cases each one, and pulmonary in 5 cases. Bone marrow infiltration was present in 13 (23.2%) cases. Twenty-nine cases were in clinical TNMstage IVB (51.7%). Thirty-six cases were reclassificated; 21 cases were excluded due to insufficient material. Twenty five cases (69.4%) corresponded to diffuse large B-cell lymphoma; of which, 48% were post-germinal origin, 40% centrofolicullar and 12% indeterminate origin. Six cases (16.6%) were reclassificated as Burkitt lymphoma; 3 of them displayed extensive plasmacytoid differentiation. Three cases (8.3%) corresponded to plasmablastic lymphomas and two cases (5.5%) were Hodgkin lymphoma. Twenty-one cases were EBV-associated (EBER positive), 19 cases were positive to NF-kB, and 12 were both positive. The global survival was of 15 months; in the cases that were EBV-positive, the average survival was of 17.3 months, and 14.5 months when NF-kB was positive. When both of them were positive, the survival decreases to 12.6 months. Conclusions: HIV-associated lymphomas are relatively frequent in our population. The prognosis is poor and the survival is short; that is the reason because is necessary to use the current and precise classification in those neoplasms. The VEB and NF-kB expression in HIV-associated lymphomas did not have significant

impact on the prognosis; nevertheless, they constitute an important mechanism in the oncogenesis of this neoplastic process. PP3-172 EPSTEIN-BARR VIRUS TUMORAL CELL EXPRESSION AND ITS IMPACT ON OVERALL SURVIVAL IN NONHODGKIN LYMPHOMAS IN TARRAGONA, SPAIN Ramon Bosch1, Marylene Lejeune1, Eva Domingo2, Joaquin Jaen1, Mª Teresa Salvado1, Barbara Tomas1, Luis Pons1, Carlos Lopez1, Patricia Escriva1, Xavier Ortin3, Marta Rodriguez3, Tomas Alvaro1, Llorenc Font3, Silvia De Sanjose4 1 Pathology Department Hospital de Tortosa Verge de la Cinta, Spain 2 Hematology Department CSU Bellvitge. Barcelona, Spain 3 Hematology Department Hospital de Tortosa Verge de la Cinta, Spain 4 Catalan Institut of Oncology, Barcelona, Spain BACKGROUND: Epstein-Barr virus (EBV) expression has been associated with an adverse prognosis in non-Hodgkin lymphomas (NHL) in few studies. AIM: To define the prognostic impact of EBV tumoral cell expression in a serie of NHL from Tarragona area in Spain. MATERIAL AND METHODS: We revised all the NHL cases diagnosed at our pathology department from the year 1990 to 2000 and we identified 265 NHL (rejecting bone marrow biopsies). After excluding the immunodeficient patients and cases without available paraffined tissue, 256 NHL cases were studied with EBER in situ hybridization. RESULTS: 24 cases (9.4%) were EBER positive (EBER+) [1 Burkitt (25%), 11 DLBC (12.36%), 1 Follicular lymphoma (2.86%), 3 small lymphocytic lymphoma (8.11%), 5 peripheral T-cell lymphomas of unspecified type (38.46%) and 3 extranodal NK/T-cell lymphoma of nasal type (100%)]. The EBER+ patients demonstrated substantially poorer overall survival (OS) [EBER+ vs. EBER-; 27 months (95% CI, 14-41 months) vs. 79 months (95% CI, 64-94 months), respectively, P=0.0013]. When studied by histological types, EBER+ remained statistically significant for poorer OS only for DLBC [EBER+ vs. EBER-; 23 months (95% CI, 3-43 months) vs. 60 months (95% CI, 47-72 months), respectively, P=0.0452] and for small lymphocytic lymphoma [EBER+ vs. EBER-; 23 months (95% CI, 12-34 months) vs. 88 months (95% CI, 59-118 months), respectively, P=0.0256]. Globally, in multivariate analysis for all LNH, EBER+ patients showed substantially poorer OS with 2.36-fold risk for death (95% CI, 1.3-4.3) while a high IPI status showed a 4.0-fold risk (95% CI, 2.6-6.0). When analysed by histological type, only the IPI status remained as a poorer independent survival factor. CONCLUSIONS: In our area, EBV expression in the tumoral cells of immunocompetents NHL patients appears to be associated with a poor OS, specially in DLBC and small lymphocytic lymphomas. However, in Cox multivariate analysis, EBV status did not affect survival with statistical significance in DLBC and small lymphocytic lymphomas. The absence of this signification could be explained, at least partially, by the small number of the EBV positive cases in this series that could affect the results. Further investigations in a larger series, in our area, are required. PP3-173 PROMOTER HYPERMETHYLATION OF GSTP1, SHP1, DAPK, p16, RB, AND p53 GENES IN DIFFUSE LARGE BCELL LYMPHOMAS: PREVALENCE AND PROGNOSTIC SIGNIFICANCE IN TUNISIAN PATIENTS Khaled Amara, Mounir Trimeche, Sonia Ziadi, Mohamed Hachana, Moncef Mokni, Badreddine Sriha, Sadok Korbi Laboratory of Pathology, CHU Farhat Hached, Sousse, Tunisia

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Background: Promoter hypermethylation is an important mechanism of inactivation of tumor suppressor genes in various human cancers including lymphomas. Diffuse large B-cell lymphomas (DLBCL) are heterogeneous in their clinical presentation, morphology, and molecular alterations. The aim of this study was to determine the prevalence and clinicopathological significance of gene promoter methylation in DLBCL in Tunisian patients. Methods: Sixty DLBCL cases diagnosed at the laboratory of Pathology at Farhat-Hached University Hospital of Sousse (Tunisia) were investigated for the methylation status of a panel of six tumor suppressor genes potentially involved in B-cell malignancies, comprising GSTP1, SHP1, DAPK, p16, RB1, and p53 by methylation-specific polymerase chain reaction. Effect of each gene methylation status on patient’s overall survival were estimated by the Kaplan-Meier method and compared with the log-rank test. Multivariable survival analyses were performed with the Cox proportional hazards model. Results: Aberrant promoter hypermethylation of DAPK, GSTP1, SHP1, p16, RB1 and p53 was detected in 75%, 70%, 65%, 53%, 20% and 3.3% of cases, respectively. All cases demonstrated aberrant hypermethylation in at least one gene. There was no significant correlation between overall gene hypermethylation status and patient’s age, gender, or clinical stage. Regarding the clinical follow-up data, available for 17 patients, promoter hypermethylation of DAPK and p16 were significantly associated with shortened survival (P=0.007 and P =0.016, respectively) and remained poor prognostic factor in multivariate analysis (P = 0.013 and P = 0.039, respectively). Conclusion: our results indicates that promoter hypermethylation is a frequent event in diffuse large B-cell lymphomas in patients from Tunisia, and that DAPK and p16 promoter regions may serve as a potential prognostic marker for predicting survival in patients with diffuse large B-cell lymphomas. PP3-174 APPENDICEAL LYMPHOMA PRESENTING AS ACUTE APPENDICITIS: A CASE REPORT Engin Cigerciogullari, Hilal Aki, Tiraje Celkan, Sergulen Dervisoglu Istanbul University, Cerrahpasa Medical Faculty, Istanbul, Turkey Malignant lymphoma comprises 1-4% of the malignant neoplasm of the gastrointestinal tract, but appendiceal lymphomas are very rare, also acute appendicitis is an extremely rare initial presenting of appendiceal lymphoma. We describe a case of a 14-year-old male, who had lymphoma of appendix presenting as acute appendicitis. His symptoms were vomitting and pain in the right lower quadrant of the abdomen. Due to the rebound and tenderness of Mc Burney’s point on physical examination, appendectomy was performed as a result of the clinical diagnosis of acute appendicitis. Histopathologic evaluation showed diffused proliferation of large and medium size lymphoma cells on mucosa and submucosa layers with acute inflamation on muscular and serosa layers. Immunohistochemical examination revealed that the lymphoma cells were positive for B cell markers. This case was diagnosed as a diffuse large cell lymphoma of the B cell type according to the WHO classification. Despite the fact that lymphoma of appendix is rare, clinically physicians and surgeons have to be aware of the differential diagnosis of acute appendicitis, because, all of cases clinically as acute appendicitis, it is not always true acute appendicitis. PP3-175 PEARSON SYNDROME: A CASE REPORT AND CLINICOPATHOLOGIC REVIEW Hilal Aki, Engin Cigerciogullari, Zeynep Karakas, Didem Apay, Gulyuz Ozturk Istanbul University, Cerrahpasa and Istanbul Medical Faculties, Istanbul, Turkey

Pearson marrow-pancreas syndrome is very rare fatal disorder that involves the hematopoietic system, exocrine pancreas, liver, kidneys in early infancy. Due to the diversity of clinical symptoms, the diagnosis can be difficult. We report on a 6 months-old boy who had presented with macrocytic anemia, neutropenia, thrombocytopenia, fever, hepatosplenomegaly and slight lactic asidosis.On bone marrow aspiration, his bone marrow was characterized by marked vacuolization of erythroblasts and myeloid precursors. Molecular analysis was made from the patient’s blood leukocytes to detect a deletion of mitochondrial DNA. When a neonate’s or infant’s bone marrow aspiration exhibits vacuolization of myeloid and erythroid precursors and also clinically the patient has metabolic acidosis of unknown cause, the possibility of Pearson syndrome should be considered. PP3-177 IMMUNOHISTOCHEMISTRY IN HODGKIN’S LYMPHOMA: DIAGNOSIS CHANGES BEFORE AND AFTER APPLICATION, TO IDENTIFY MORPHOLOGIC AND BIOLOGIC GREY ZONE LYMPHOMAS Nuray Bassullu1, Nukhet Tuzuner2 1 Departments of Pathology, Ministry of Health, Antakya Goverment Hospital, Hatay, Turkey 2 Departments of Pathology, Cerrahpasa Medical Faculty, Istanbul, Turkey BACKGROUND: There is a diagnostic grey zone between Classic Hodgkin lymphoma (CHL) and some histologic types of non-Hodgkin lymphoma (NHL), including anaplastic large cell lymphoma (ALCL) , T-cell-rich B-cell lymphoma (TCRBCL) and mediastinal large B cell lymphoma. Some morphological and/or phenotypic features are common in both CHL and those particular NHL types. METHOD AND RESULT: In this study, the cases of HL, diagnosed and subtyped without immunohistochemistry (IHC) were reinterpreted after the performance of IHC and the cases shareing common features with NHL were isolated. In addition the cases confused with HL in the morphological and/or biological grey zone but diagnosed after IHC as ALCL and TCRBCL were taken in the study group and together with HL, and all were reevaluated. After the reinterpretation of 508 cases, the diagnosis of CHL was reestablished in 390 cases out of 429 (90,9 %).In 35 cases (8,1 %) the diagnosis was changed to NHL. NHL subtypes were ALCL in 11 cases,TCRBCL in 21 cases, PTCL (peripheral T-cell lymphoma) in 1 case, DLBCL(diffuse large B-cell lymphoma) 1 case and B cell ALCL in 1 case. In 2 cases the differentiation between HL and TCRBCL, in 2 cases the differentiation between ALCL and HL couldn’t be achieved. Out of 11 cases in which the differential diagnosis between HL and NHL couldn’t be achieved initially, the diagnosis was established as ALCL in 3 cases(27,3 %) and as HL in 7 cases (63,6%) after the reevaluation of the cases. In 1 case the differentiation between NLPHL and TCRBCL couldn’t be made. In 31 cases of TCRBCL,the initial diagnosis wasn’t changed. Out of 37 cases initially diagnosed as ALCL the differentiation between ALCL and HL couldn’t be achieved in 1 cases . CONCLUSION: The grey zone between HL and NHL can either be a biological grey zone as a result of common pathogenesis or it may be a morphological grey zone due to the morphological similarities.In conclusion, as the biology and the origin of RS cells were cleared out, the differentiation of HL from other aggressive lymphomas such as TCRBCL or ALCL became possible at least to an extend. However, in cases displaying CD 15 negativity and high ratios of CD20 positivity, the differential diagnosis was still troublesome.

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PP3-178 CLINICAL ROLE OF FLOW CYTOMETRY IN REDEFINING BONE MARROW INVOLVEMENT IN DIFFUSE LARGE B-CELL LYMPHOMA (DLBCL) – A NEW PERSPECTIVE Dipti Talaulikar1, Bruce Shadbolt2, Jill Bell3, Kowsar Khan3, Jane Dahlstrom4 1 Department of Haematology, ACT Pathology, The Canberra Hospital and Australian National Univeristy Medical School, Canberra, ACT, Australia 2 Department of Epidemiology, The Canberra Hospital and Australian National Univeristy Medical School,Canberra, ACT, Australia 3 Department of Haematology, ACT Pathology, The Canberra Hospital, Canberra, ACT, Australia 4 Department of Anatomical Pathology, ACT Pathology, The Canberra Hospital and Australian National Univeristy Medical School, Canberra, ACT, Australia Aims: The clinical role of flow cytometry in staging bone marrows in Diffuse Large B-cell Lymphoma (DLBCL), especially its impact on outcome, remains ambiguous. The aims of this study were to determine the contribution of flow cytometry to conventional staging, and to study impact on survival. Methods and results: One hundred and thirteen cases of DLBCL diagnosed at The Canberra Hospital from 1996-2005 were identified for this retrospective study. On blinded analysis of Bone marrow (BM) aspirates, trephine biopsy slides, and flow cytometry data, involvement on morphology (M) was detected in 23 (20.3%) cases and on flow cytometry (F) in 25 (22.1%) cases with discordance noted in 18 cases (15.9%). Together, the 2 diagnostic techniques (M + F) detected 33 cases (29.2%). Cases with and without marrow involvement on conventional staging alone (M) had no significant difference in survival (p=NS). However, when bone marrow involvement was defined as positivity on morphology and/ or flow cytometry (M + F), the median survival of patients with involvement was significantly worse than patients without involvement (p=0.02). Conclusions: Flow cytometry positive cases should be included in a summative model to define bone marrow involvement in DLBCL as it may have a potential impact on outcome. PP3-179 IMMUNOHISTOCHEMICAL EXPRESSION OF CD34 IN LYMPH NODE AND BONE MARROW OF PATIENTS WITH HODGKIN’S LYMPHOMA Tatjana Terziü1, Vesna ýemerikiü –Martinoviü2, Maja Peruniþiü-Jovanoviü3, Ljubomir Jakoviü3, Sanja Radojeviü-Škodriü1, Gordana Basta-Jovanoviü1 1 Institute of Pathology, School of Medicine, University of Belgrade, Serbia 2 Histolab, Belgrade, Serbia, 3 Institute of Hematology, Clinical Center of Serbia, Belgrade, Serbia Background: Bone marrow infiltration is present in 5-15% of patients with Hodgkin’s lymphoma (HL). It is usually associated with prominent fibrosis and proliferation of blood vessels. Limited information exists about the significance of neoangiogenesis in HL. In this study we analyzed immunohistochemical expression of CD34 in lymph node and bone marrow of patients with HL. Methods: We identified 39 patients with HL (25 male and 14 female, median age 43 years), 25 patients with bone marrow infiltration by HL and 14 patients without bone marrow infiltration. Slides from paraffin embedded lymph node and bone marrow biopsy were stained with hematoxylin&eosin and reticulin fibers (Gordon-Sweet). Microvessels were visualized by immunohistochemical staining for CD34. We counted the number of vessels per high power field (x500) in two the most vascularized areas. Results: Microvessels density (MVD) in lymph node with HL was lower than in reactive lymph node (37±1,73). There was statistically

significant difference (p<0,05) between MVD in lymph node of HL patients with bone marrow infiltration (14,75± 5,23) and MVD in lymph node of HL patients without bone marrow infiltration (20±3,95). In patients with HL, but without bone marrow lymphoma infiltration, we found increase of MVD in bone marrow (9,15±5,06), compared with control bone marrow (3,50±0,71), but correlation was not statistically significant (p=0,205). We observed that MVD inside the bone marrow lymphoma infiltrate (17,63±9,27) in HL patients was higher than MVD in HL patients without bone marrow infiltration (9,15±5,06). This correlation was statistically significant (p=0,03). Conclusion: Lower MVD in lymph node of HL patients than MVD in reactive lymph node suggests that lymphoma proliferation leads to the destruction of the existing vessels rather than neoangiogenesis within lymph node. But, our findings suggest that neoangiogenesis in bone marrow infiltration by HL was extensively increased. Therefore, the density of microvessels may provide useful prognostic information and basis for the therapeutic investigation of anti-angiogenic agents in the cases of HL with bone marrow infiltration. PP3-180 THE DIFFERENTIAL DIAGNOSIS OF MAST CELL PROLIFERATIONS ON FIVE REPRESENTATIVE CASES Cevriye Cansiz, Gulsah Kaygusuz, Eda Akpinar, Isinsu Kuzu Ankara University, School of Medicine, Department of Pathology, Ankara, Turkey The term mastocytosis denotes a heterogenous group of disorders characterized by abnormal growth and accumulation of mast cells (MC) in one or more organ systems. It subdivides into the indolent systemic mastocytosis (ISM), Systemic mastocytosis with an associated clonal hematologic non-mast cell lineage disease (AHNMD), aggressive systemic mastocytosis (ASM) and mast cell leukemia (MCL). It is important to differentiate the mast cell diseases from the cytokine related mast cell hyperplasia in the bone marrow underlying other inflammatory processes or tumors without typical skin lesions or organ involvement. Demostrating the missense mutations on c-Kit gene is becoming important for astimating result of targeted drugs for treatment modalities. Our aim in this study is to discuss the diagnosis of different types of mast cell involvements in the bone marrow of 5 cases with their clinical and pathological presentations. Three of these cases were fulfulling the criteria of ASM with multiorgan involvement. One of these cases couldn’t be diagnosed for 5 years although she had splenectomy 3 years ago.Differing from the other SM cases the skin involvement could not be demonstrated in this case and her skin complaints were accepted as secondary symptom related to histamine effect. Two of the cases were fulfilling the criteria of AHNMD. One of these cases had cured Hodgkin’s Lymphoma history diagnosed 13 years ago and cured secondary acute myeloid leukemia (AML) history diagnosed 2 years ago. The bone marrow was in remission for AML but there were focal paratrabecular infiltration of spindle shaped mast cells fulfilling the criteria of atypical type 1 mast cell infiltration. The mast cells were not recognised on the first two bone marrow biopsies taken for the remission control of AML. The second AHNMD case was Waldernstroms macroglobulinemia developed under the treatment of rheumatoid artritis. The bone marrow was interstitionally infltrated by monoclonal lymphoplasmocytic B cells and multifocal paratrabecular groups of mast cells. The mast cell infiltration became more clear and increased on the second biopsy taken for her disease follow up. The mast cell population were confirmed by immunohistochemistry for every case by their positive staining with CD117, CD68 and mast cell tryptase. Conclusion: Neoplastic mast cell proliferations should be diagnosed and classified correctly for performing the appropriate treatment modality. And should be differentiated from secondery reactive mast cell proliferations in order to avoid unnecessary toxic treatment.

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PP3-181 THE RELEVANCE OF T CELLS AND FOLLICULAR DENDRITIC RETICULUM CELLS ON PREDICTING THE CLINICAL BEHAVIOUR OF FOLLICULAR LYMPHOMAS Ayse Ozgun1, Duygu Enneli Kankaya1, Sare Altas2, Gulsah Kaygusuz1, Sule Mine Bakanay3, Muhit Ozcan3, Isinsu Kuzu1 1 Ankara University, School of Medicine, Department of Pathology, Ankara, Turkey 2 Ataturk University, School of Medicine, Department of Pathology, Erzurum, Turkey 3 Ankara University, School of Medicine, Department of Hematology, Ankara, Turkey Follicular lymphomas (FL) are most commonly seen in western populations (22-35 %) whereas their incidence is around 8-10% in Turkish population. Recent studies including large series of FL cases revealed that the well known histopathological prognostic criteria may not be enough for estimating their clinical behaviour. This situation has brought the idea of focusing on the characteristics of microenvironment besides neoplastic cells. Our aim in this study is to compare the microenvironment and proliferation status of follicular lymphomas with known prognostic histopathological criteria. Methods: 33 follicular lymphoma patients (16 female, 17 male) with a mean age of 54,5 (28-75), diagnosed between the years 2002-2007, were included in this study. All cases were reviewed by examining their histological and immunohistocemical features. The WHO criteria was used for histological grading. As well as the known prognostic histopathological parameters, the presence of residual reactive follicules, the preservation of follicular dendritic reticulum cell (FDRC) network, the density of T lymphocytes and histiocytes within the follicules and the proliferation status of the neoplastic follicules by Ki-67 staining were evaluated. Chi square, Mann Whitney U, Kruskal Wallis tests were used for statistical analysis. Results: Eight cases were graded as grade 1, 12 cases as grade 2, 13 cases as grade 3. There was statistically significant correlation between histological grade and Ki-67 index (p<0.05), while such correlation was not present between the histological grade and the bcl-2 expression, the density of CD3 positivity and the ratio of diffuse areas. The cases having higher proliferation index (>49%) showed significantly decreased density of reactive T cells (p=0,01). No correlation was found between Ki-67 index, the preservation of FDRC network and the expression of bcl-2. Conclusion: Our results suggested that the proportion of reactive T lymphocytes within the neoplastic follicules may predict the biological behaviour of FL. In according to our data, preservation of FDRC network is not associated with histological grade, proliferation status or T cell population in FL.

Skin Pathology PP3-182 FIVE YEARS STATITICAL ANALYSIS OF HENOCH SCHOENLEIN PURPURA AND IT’S RELATION WITH SEASONS Fakhrozaman Pezeshkpour, Ramin Ghasememi, Mohammad Javad Yazdanpanah, Hamid Ahanchian Ghaem hospital, Department of Dermatology, Mashhad university of Medical sciences, Mashhad, Iran Abstract: Introduction and purposes: HS purpura is the most distinctive subgroup of hypersensitivity vasculitis that is characterized by purpura in buttocks and lower extremities, involvement of kidney, joint and abdomen. The main aims of this study include: Determination of the correlation between HS purpura frequency and seasons. We also determined the distribution of age and sex and frequency of HS purpura. Methods: This descriptive study was done between1993 to1998 in patients referred to dermatology clinic and pediatric emergency of Ghaem hospital. The diagnosis was based on clinical and pathological findings. Results: There were 40 patients, with M/F ratio 0.8. 13 patients (32.5%) presented in spring, 7 (17.5%) in summer, 11 in autumn (27.5 %) and, (22.5%) in winter. All of the patients had typical skin lesions, 57.5 % in lower extremity 22.5% in upper and lower extremity, 17.59 in upper, lower extremity and trunk. And 2.5% only in upper. The most frequent non cutaneous lesion belongs to abdominal and joint involvements (39%). Conclusion: The most frequent seasonal distribution was in spring and the least frequent was in summer and there was a significant differences between the two seasons (p=0.05). But there was not such significant difference among all seasons. The most frequent site of skin lesions was the lower limbs (97.5%). PP3-183 FEED FORWARD NEURAL NETWORK AS A DIAGNOSTIC HELP IN IMMUNOFLOURESCEIN PATTERNS OF SKIN DISORDERS Sulen Sarioglu1, Mustafa Sakar2, Banu Lebe1 1 Dokuz Eylul University Faculty of Medicine Department of Pathology, Izmir ,Turkey 2 Dokuz Eylul University Faculty of Medicine Department of Clinical Engineering, Izmir, Turkey BACKGROUND: Examination of the tissues by microscopy still depend upon the visual cognitive abilities of the pathologists. Although automatisation seems to be far away, image analysis methods are in progress. Measurements of area, volume and stained area percentage methods by image analysis may be applied especially for determining prognostic factors. Neural networks are mathematical algorithms that depend upon the functionality of small neural clusters. The study is designed in order to find if Feed Forward Neural Network (FFNN) depending upon automatic image analysis measurements in DIF images from skin disorders might help the pathologist. METHOD: Images from anti IgG, IgA, IgM, C3 and fibrinogen DIF stained sections from the skin biopsies from patients with immunodeposition were collected to a computer from immunoflourescence microscope. The patterns of deposition were grouped as: basement membrane linear (BML), basement membrane granular (BMG), epidermal intercellular (IC), vascular (V), negative and nonspecific background staining. Variables that might be helpful in the differential diagnosis were evaluated and total size, node, link, end point and branch were selected. The training set included 193 images from 32 cases with positive DIF results. An experienced pathologist suggested a possibility range for each image about the 6 categories. After the training process of FFNN the values for each variable were analyzed statistically for comparing the groups. For determining the automatic self made decision process based on learning, 100 images from 40

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cases were analyzed by FFNN. The decisions of the pathologist and the category with highest possibility level selected by FFNN were compared. RESULTS: All the variables were significantly different when all the groups were considered (One Way Anova p<<0,000), when groups were compared separately, all values for all variables for IC pattern was significantly different from other groups, but the others gave mixed results. Of the 100 images FFNN gave the highest possibility in 83% images, in concordance with the pathologist, but gave the right result as the second possibility in 14 cases. For the IC pattern best results were obtained (89% first possibility) followed by V (85%), BML (84%) and BMG (76%). CONCLUSIONS: DIF image FFNN may help inexperienced pathologists in future. Selection of total size, node, link, end point and branch as variable seems valuable. The FFNN may provide better results with long term training. PP3-184 EVALUATION OF MUTANT p53 GENE FREQUENCY IN PATHOLOGIC SAMPLES OF PATIENTS WITH NONMELANOTIC SKIN CANCERS AND ITS CORRELATION WITH CLINICOPATHOLOGIC ASPECTS Mahnaz Banihashemi, Fakhrozaman Pezeshkpoor, Norieh Sharifi Qaem Hospital, Iran Introduction: We aimed to investigate frequency and immunoreactivity of mutant p53 gene in non-melanotic skin cancers (NMSC) and to correlate its Over-expression with clinicopathologic aspects. Methods: Sixty paraffin-embedded pathology blocks with NMSC registered Diagnosis, including 39 BCC and 21 SCC were analyzed immunohistochemically To assess over expression of p53 protein related to sun-exposure ,age and gender Not only have we divided the samples to positive mutation samples and negative Mutation ones ,we sorted samples in the quality method to 0(negative), +1(mild), +2(moderate) and +3(severe) for p53 mutation based on visual impression and severity of staining. We also quantified proportion of the cells expressing p53 within a tumor; obtaining the percentage of the stained nuclei. Result: The frequency of mutant p53 gene in BCC and SCC was 82.1% and 71.4% respectively. Age and gender did not affect over expression of p53 significantly Frequency of mutant p53 gene was 89.1% among sun-exposed sites and 42.9% among unexposed sites to UV radiation (p<0.001). When sunexposed effect was studied in BCC and SCC separately; 87.1% of sun-exposed BCC showed p53 mutation (P=0.137)while 93.3% of sun-exposed SCC significantly showed p53 mutation (P=0.002). however ,sun exposure of the tumor site increases the severity of p53 staining-when mutation existed-in SCC and BCC significantly (P=0.015 & 0.005 respectively). Also, exposure to ultra violet radiation increases the mean of percentages of stained nuclei-when mutation existed- in SCC and BCC significantly (P=0.04 & 0.008 respectively). Conclusion: Chronic UV radiation can\\\'t induce p53 mutation in BCC but when the mutation occurs due to any other factor, in increases the quality of the activity of mutated p53 gene. In SCC chronic exposure to UV induces mutation in TP53 and also it increases the activity of mutated gene in both quality and quantity. PP3-185 THE EFFECT OF POST-BURN LOCAL HYPERTHERMIA ON THE REDUCING BURN INJURY: THE POSSIBLE ROLE OF OPIOIDS Shahram Shahabi1, Mahmud Hashemi2, Zuhair Muhammed Hassan2, Behrouz Ilkhanizadeh3, Nima Hosseini Jazani1 1 Department of Immunology and Microbiology, School of Medicine, Urmia Medical Sciences University, Urmia, Iran 2 Department of Immunology, School of Medical Sciences, Tarbiat Modarres University, Tehran, Iran. 3 Department of Pathology, School of Medicine, Urmia Medical Sciences University, Urmia, Iran

Purpose: This paper studied the effect of post-burn local hyperthermia on burn induced injury. Methods: A second-degree burn injury was induced on the right and left flanks of Balb/c mice.Thirty-two burn wounds were divided into four groups. Opioid receptor blocking was done for groups 3 and 4 by intraperitoneal administration of Naloxone (NLX) 30 min before the thermal injury. Local hyperthermia (45°C, 30 s) was applied only for the peripheral area of burn wounds of groups 2 and 4. Twenty-four hours after burn injury, the burned wounds were assessed for the level of iNOS (by immunohistochemistry) and the number of hair follicles (as an indicator of tissue injury). Results: The wounds that received hyperthermia (group 2) had significantly more hair follicles (p < 0.001) compared to the control wounds (group 1). There was no significant difference between the number of hair follicles and acute inflammation of group 1 and group 3 (NLX + burn). Group 4 (NLX + burn + hyperthermia) had significantly fewer hair follicles compared to group 1 (p < 0.001), group 2 (p < 0.001) and group 3 (p < 0.001). The level of iNOS in groups 1, 3 and 4 was not significantly different but significantly more than group 2 (p < 0.001, p < 0.001 and p < 0.001, respectively). Conclusions: The results showed that local hyperthermia after second degree burn decreased the tissue injury and iNOS expression. It is also concluded that endogenous opioid response may have a key role in the above mentioned effects of post-burn local hyperthermia. PP3-186 CASE OF THE BROOKE-SPIEGLER SYNDROME Pouran Layegh1, Nourieh Sharifi Sistani2, Fakhrozaman Pezeshkpour3, Mohammad Abadian3 1 Dermatology Department,Qaem Hospital, Mashad University of Medical Sciences, Iran 2 Pathology Department,Qaem Hospital, Mashad University of Medical Sciences, Iran 3 Dermatology Department, Qaem Hospital, Mashad University of Medical Sciences, Iran Case Report : The patient is a 26-year old woman, presented with cutaneous lesions on her scalp and face. These lesions had first appeared when she was 14 years old and primarily on her scalp which later developed on the face and had gradually increased in size and number over the years. In the patient's family, her father and brother, who was 13 years old had similar lesions on their faces. On examination, a group of round to oval –shape skin – coloured papules with a smooth pearly surface measuring 2-6 mm in diameter were seen in the mid-face, particularly in the nasolabial folds, the upper lip and less commonly on the forehead and the periocular areas.In addition to this, a variety of tumoral and nodular lesions was also seen on the scalp. The tumors of the scalp were pinkish –red, dome-shaped to some extent peduncular with surface telangiectasia and induration on palpation. They were measured 0.5-1.5 cm in diameter, the larger lesions were hairless and the smaller ones had less hair than normalAlso there was a subcutaneous firm nodule with a diameter of 0.5 ×0.5 cm on the patient´s right forearm. Her general condition was good; she had no known underlying disease and no previous history of taking any kind of medications. The histopathology examination of the scalp nodule revealed a dermal lesion with defined borders composed of islands of basaloid cells arranged in a jigsaw puzzle-like pattern and separated by hyaline basement membrane material. Two populations of basaloid cells (with a large nucleus centrally and smaller nuclei in a palisading pattern of the periphery) were observed, which can represent Cylindroma. In the facial papules basaloid inclusions with immature (basic) follicular papillae and horncysts were seen which were surrounded by fibrous stroma and are typical of trichoepithelioma .The patient's forearm nodule composed of sheets of small dark staining basaloid cells peripherally and larger cells with paler nuclei centrally, consistent with spiradenoma According to the clinical feature and the histopathology findings, the diagnosis of was Brooke _Spiegler syndrome made.

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PP3-187 HISTOLOGICAL ANALYSIS OF HUMAN SKIN FROM EGYPTIAN MUMMIES Dina Tiniakos1, Norah Caldwell1, Vasso Zafiri2, Athanassios Sakarellos2, Christos Kittas1, Theodoros Pitsios2 1 Laboratory of Histology & Embryology, Medical School, National & Kapodistrian University of Athens, Greece 2 Museum of Anthropology, Medical School, National & Kapodistrian University of Athens, Greece Background. Four Egyptian mummies of the Hellenistic/Roman period (332 B.C.-395 A.D.), an adult man, an adult woman and two young male children, are included in the ethnologic collections of the Museum of Anthropology, University of Athens, Greece. The biological age of the mummies, based on anthropometric and dental studies, is estimated at 25-35, 40-50, 7-8 and 2.5-3 years, respectively. Anatomical and imaging (xRay, computed tomography scan) studies combined with 3-D virtual reconstruction of scull and bones were undertaken. The aims of the present study, which to our knowledge is the first histological study on Egyptian mummies in Greece, were to optimize the conditions of mummified tissue specimen preparation for histological analysis using light microscopy and to evaluate the preservation state of tissue components. Method. Samples of skin tissue from all four mummies were taken without damaging the remains. Four different methods of tissue rehydration were applied for specimens from each mummy using a) 10% sodium bicarbonate in distilled water (dǾ2ȅ), b) fabric softener, c) 5% glucose in dǾ2ȅ and d) 3% dimethylsulfoxide in dǾ2ȅ, followed by fixation in 10% neutral formalin and embedding in paraffin. Serial 5 m- thick tissue sections were cut from each paraffin block and the histochemical stains hematoxylin-eosin and Masson Trichrome were performed. Results. Immersion of the specimens in 10% sodium bicarbonate solution for 8 hours was considered the optimal (most rapid and with best tissue preservation) rehydration method for the desiccated mummy skin. Histological analysis using light microscopy showed absence of epidermis in all tissue sections examined, while the dermis appeared well preserved with identifiable collagen fibers in the dense, irregular fibrous tissue which stained lightly with Masson Trichrome stain. No visible cells or cell remnants were present. Conclusions. We performed a, unique in Greece, histological analysis of 1700-2300-year-old human skin from Egyptian mummies. Rehydration in sodium bicarbonate solution followed by routine fixation and embedding was considered the best method for morphological and histochemical analysis of mummified human tissues. Collagen fibres and possible blood vessels were identified in the dermis of the mummified skin. PP3-188 A STUDY OF MONOCYTIC AND DENDRITIC CELL MARKERS IN BENIGN CUTANEOUS FIBROUS HISTIOCYTOMA (DERMATOFIBROMA) Daisuke Nonaka, Luis Chiriboga New York University School of Medicine, USA Background: Controversies have existed regarding the histogenesis of fibrous histiocytomas (FHs) as well as the issue regarding whether they represent a neoplastic or reactive process. Based on the expression of factor XIIIa (fXIIIa), the proliferating cells of these lesions have been considered to be dermal dendrocyte by some investigators. We report on the expression of various novel monocytic and dendritic cell markers on dermatofibromas (DFs) and, for comparison, giant cell tumor of tendon sheath (GCTTS). Design: A total of 38 cases of DF and 10 cases of GCTTS were stained with CD1a, CD11c, CD68, CD83, CD123, CD205 (DEC-205), CD207 (Langerin), CD208 (DC-LAMP), CD209 (DC-SIGN), fXIIIa, muscle specific actin (HHF-35), smooth muscle actin (SMA), and calponin. Results:

CD163, CD68 and fXIIIa were diffusely expressed in approximately two-thirds of the DFs, with the remaining cases being focally positive; myeloid dendritic cell markers including CD83, CD205, and CD208, and Langerhans cell markers such as CD1a and Langerin were negative; plasmacytoid dendritic cell marker, CD123, was completely negative; SMA and calponin were variably expressed in half of the cases; and HHF-35 was focally stained in only two cases. CD11c and CD209, which can be expressed in both myeloid dendritic cells and macrophages, were focally positive. Both CD68 and CD163 were diffusely stained in GCTTSs; fXIIIa was variably expressed in 8/10 cases; dendritic cell markers were negative; and smooth muscle markers were also negative except for focal staining of SMA in one case. CD11c and CD209 were also focally expressed. Conclusion: DFs are composed predominantly of cells with macrophage markers with a variable amount of myofibroblastic elements, without evidence of specific dendritic cell phenotype. GCTTSs are composed of cells of macrophage derivation. Our results support the notion that monocyte/macrophage represent the potential histogenesis of DFs. PP3-189 HISTOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL ASPECTS IN FEW TYPES OF CUTANEOUS SARCOMAS Costache Mariana1, Simionescu Olga2, Ene Ana-Maria3, Sajin Maria1 1 V. Babes National Institute of Pathology, Romania 2 UMF Carol Davila , Romania 3 Faculty of Biology University of Bucharest, Romania The present study describes the histopathological and immunohistochemical aspect in different cases of cutaneous sarcomas which are representative and also rare in the medical practice (dermatofibrosarcoma protuberans, malignant fibrous histiocytoma, leyomiosarcoma, rhabdomyosarcoma, hemangiopericytoma, malignant hemangioendothelioma ). Material and method: We have done a retrospective study within a period of two years concerning on a group of 35 patients who presented cutaneous sarcomas. We used surgical excision pieces, which had been fixed in buffered 10% formalin, paraffin embedded and sectioned at 3ȝm, then stained routinely by Hematoxylin - Eosin was used in the majority of cases and special stains (Gomory - silver impregnation) in several others, than examined by light microscopy. For the immunohistochemical (IHC) staining the following antibodies were used: CD34, CD31, Actin, Myoglobin, Desmin, Cytokeratin, Vimentin. Results:The primary cutaneous sarcomas have a various neoplastic forms; can occur at any age; these neoplasms to children are rare. Both sexes are affected. As an essential and distinctive feature of these tumours there is a risen tendency of recur. Clinically,the most sarcomas appear solitary and have a profound invasion; the usual locations are the extremities and the retroperitoneal region. The lesions are present as nodules or plaques. Frequently, in ultimate stages, the tumour may necrosise and ulcerate. Histopathologically, they are characterized by invazivity, being not encapsulated and unpreciselly delimitate by the neighbouring tissues. The cellular and nuclear atypia of size, form and colour, as well as atypical mitoses are frequently present. The tumours prouve high cellularity with the tendency of desposing the cells in different oriented fascicles. The vascular tumours show irregular vascular lumens surrounded by neoplastical proliferations. Immunohistochemical staining showed a diffuse positivity for vimentin; CD34 and CD31 was positive in vascular structures; cytokeratin was negative and actin and myoglobin were positive in tumour cells and in vassels walls. The cutaneous sarcomas represents a heterogeneous group of tumours with many histopathological aspects. However, a large group of them consists in problems of positive and differential diagnosis. The

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immunophenotyping of such a tumour is important because in correlation to the anatomo-clinical features permits a good differential diagnosis and rules out a large number of tumoral lesions. PP3-190 LANGERHANS CELLS AS A CLUE FOR TUMOROUS NATURE OF DERMATOFIBROMA Ivana Iliü, Zdenka Hutinec, Jadranka Iliü-Forko University Medical Center Zagreb, Serbia Background. Dermatofibroma is a common benign lesion usually found on extremities and trunk of adults. Despite its frequency and benign nature it is still unknown whether this is an inflammatory lesion or a true neoplasm. In most inflammatory lesions of the skin the number of epidermal Langerhans cells is higher than in normal epidermis. The number of epidermal Langerhans cells in the dermatofibroma could be a useful, although indirect, clue for defining whether this entity is closer to the inflammatory lesions or to the neoplasms. Method. In our study we included 10 patients with dermatofibroma, 10 with dermatofibrosarcoma protuberans and 10 with postoperative scar. All lesions were immunochistochemicaly stained with CD1a (Clone O10, dilution 1:50, Dako Cytomation, Glostrup, Denmark) to highlight Langerhans cells. We counted the number of Langerhans cells in the epidermis above the lesion. The intraepidermal CD1a positive cells were enumerated as a percentage of 100 keratinocytes. Two concomitant areas of high power field were counted and mean +/-SD percentage was calculated in each specimen. For comparing the differences between mean number of epidermal Langerhans cell in scar, dermatofibroma and dermatofibrosarcoma protuberans we used Student t-test. p <0.05 was regarded as statistically significant. Results. By comparing the number of Langerhans cells (mean+/SD) in epidermis above the scar (7.62+/-1.38), dermatofibroma (2.05+/-0.61) and dermatofibrosarcoma (2.76+/-1.31), we found that there is no statistically significant difference between mean number of epidermal Langerhans cells above the dermatofibroma and dermatofibrosarcoma (p=0.160). The difference between means of epidermal Langerhans cells number above dermatofibroma and above scar is found to be statistically significant (p<0.0001). Conclusion. This was an indirect way of showing the neoplastic nature of dermatofibroma. If dermatofibroma was an inflammatory lesion, we expected dermatofibroma to have higher number of epidermal LC, just like we found in scars. PP3-191 DOES MERKEL CELL CARCINOMA REPRESENT AN EPITHELIAL TUMOUR WITH NEUROENDOCRINE DIFFERENTIATION Vuka Katic, Jasmina Gligorijevic, Katarina Katic, Boban Stamenkovic Institute of Pathology, Medical Faculty Nis, Serbia Background: Merkel cell carcinoma (MCC) is uncommon malignant skin tumour with a poor prognosis and rapid progression. Most MCCs occur in elderly individuals, on sunexposed areas of the body, with the head and neck being the most common sites, followed by the extremitaties and trunk. However, the origin of the MCC is still controversial, because its cells have both epithelial and neuroendocrine elements. To answer on this dilemma we have investigated an expression of neuroendocrine and epithelial markers in MCC. Aims: The origin of the Merkel cell carcinoma is still controversial because its cells have both epithelial and neuroendocrine elements. To answer on this dilemma we have investigated an expression of neuroendocrine and epithelial markers in Merkel cell carcinoma. Methods: The study group was composed of 6 cases, with the mean age of 68 years. Multiple subcutaneous masses and lymph node metastases

were surgically ressected.Paraffine blocks of the formaldehydefixed tumorous tissue were cut and stained with H&E, PAS, HIDAB pH=2,5 and LSAB2 techniques.The following antibodies (Dacopatt) were used: Chromogranine A, Neurone specific enolase (NSE), panCytokeratin, Melan A, CD20 and CD45Ro. Results: The tumours involved the dermis, with sparing of the epidermis. A variety of histological subtypes were recognized, including intermediate, small cell, mixed intermediate and smallcell variantsMitoses were tipically numerous, and atypical forms were frequently seen. Lymphovascular invasion was extrmely common finding. MCCs expressed both neuroendocrine and epithelial differentiation. They showed positive labeling with epithelial membrane antigen, dot paranuclear panCytokeratin, as well as positive labeling with Chromogranine A and NSE, markers for neuroendocrine differentiation. Conclusion: Merkel cell skin carcinoma is immunohistochemically epithelial malignant tumour with neuroendocrine differentiation. PP3-192 TUMOUR REGRESSION, A VARIABLE TO BE AWARE OF IN EVOLUTION OF CUTANEOUS MELANOMA Ana Fuertes1, Carlos Horndler1, Ramiro Álvarez1, Maria Teresa Puértolas2, Isabel Marquina1, Mar Pascual1, Guillermo Muñoz1 1 Pathology Department, Miguel Servet University Hospital, Zaragoza, Spain 2 Oncology Department, Miguel Servet University Hospital, Zaragoza, Spain BACKGROUND: Prognostic variables of melanoma in TNM classification (AJC 2001) are Breslow thickness, ulceration of epidermis and Clark level. Besides, some other variables have been considered such as regression and mitotic index. Malignant melanoma is one of the tumours with a highest rate of spontaneous regression, around 15% of all cases. Regression has been associated with an increase of angiogenesis and a higher risk of developing metastases. METHODS: We have studied 116 cutaneous melanomas, of which we had all clinical and outcome data along 3 years at least. We have considered histological variables such as Breslow thickness, Clark level, presence of ulceration, histologic type, mitotic index, lymphoid response and tumour regression. We have also performed an immunological study of beta-catenin, Ki-67, Bcl2, p16, EGFR, c-Kit and cyclines (B1, D1 and D3). RESULTS: Descriptive study shows regression in 15 of the 116 studied patients (12,9%). In univariate analysis there are differences in survival-without-recurrence rate, being shorter the rate of patients with tumour regression (p=0,0073). Multivariate analysis shows that recurrence rate is 5,6 times higher in patients with tumour regression, and risk of death is 3,51 times higher for these patients. Presence of regression is associated with a higher risk of recurrence and a shorter specific survival for melanoma. CONCLUSIONS: Most bibliographic data seem to indicate that regression is not a significant prognostic factor, but it is related to an increase of angiogenesis, and therefore with a higher capacity of distant dissemination. In many studies regression does no seem to be a significant prognostic factor, probably because it is difficult to be defined and assessed. Other studies affirm that it can be an important factor, especially when Breslow thickness is less than 1 mm, and most of all when regression is extensive. In our case series, 33,33% of the patients with regression areas had a Breslow of 1 mm or below, and presence of regression correlates with an increase of the risk of recurrence and death. PP3-193 ASSESSMENT OF THE DEMODEX FOLLICULORUM IN HUMAN CUTANEOUS LESIONS Narcisa Mederle1, Ovidiu Mederle2, Marius Raica2 1 Veterinary Medicine Faculty, Timisoara, Romania 2 University of Medicine and Pharmacy, Timisoara, Romania

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Introduction Since the epizootic conditions of the appearance of dermatoses are common and the clinical signs often lead to confusions, a skin smear, correctly executed and interpreted, is helpful in the diagnosis of demodicosis and in the differential diagnosis between skin lesions. Cited by many authors as a saprophyte of the human skin, Demodex Folliculorum is the cause of many skin lesions, such as folliculitis, desquamations, prurit, hyperseborrhea of the facial region. Material and method 14 patients (3 men and 11 women) diagnosed with different dermopathies (endocrine, allergic, bacterial) which were non – responsible to the usual therapy were examined. The patients had erythema, folliculitis, desquamations, hyperseborrhea of the facial region. The skin smears were clarified in lactic phenol and paraffin oil and were microscopically examined. From the positive smears we have made measurements of 31 adult acarians. Results The microscopic exam of the skin smears showed the adult form of Demodex Folliculorum in all the patients. No evolutive stages of the parasite (egg, larva, nymph) were found. Medium sizes of the female and male parasites were 238/47 ȝm and 212/46 ȝm, respectively. The explanation for the absence of the intermediate stages of the parasite is that the samples were taken from the superficial skin, careful not to produce supplementary lesions or to leave scars (especially in women). Conclusions The microscopic exam of the skin smears of 14 patients diagnosed with dermatoses with other aetiology than parasitic and non responsible to treatment has shown the presence of the adult form of Demodex Folliculorum. The morphologic description and the dimensions of the adult Demodex Folliculorum were in correlation with bibliographic data. PP3-194 EXPRESSION OF HPV, p53, Bcl-2 AND PCNA IN PREMALIGNANT AND MALIGNANT EPIDERMAL TUMORS George Burkadze1, Oleg Kvlividze2, Gulisa Turashvili1, Liana Gogiashvili2 1 Department of Clinical Pathology, N. Kipshidze Central University Clinic, Tbilisi, Georgia 2 A. Natishvili Institute of Experimental Morphology, Tbilisi, Georgia Background. Bowen’s disease is an intraepithelial squamous cell carcinoma which has been related to human papillomavirus (HPV) infection. Non-melanoma malignant skin tumors such as invasive squamous cell carcinoma and basal cell carcinoma have also been associated with HPV. The interaction between ultraviolet light and HPV infection has been suggested to be a major mechanism for skin carcinogenesis. The purpose of our study was to assess the immunohistochemical characteristics of Bowen`s disease and invasive epidermal tumors. Methods. We examined immunocompetent patients with Bowen`s disease and malignant epidermal tumors such as invasive squamous cell carcinoma and basal cell carcinoma. Ten patients were involved in each group. Biopsy specimens were fixed in 10% neutral buffered formalin and embedded in paraffin. The paraffin sections were stained by hematoxylin-eosin and von Gieson. Immunohistochemistry was performed using monoclonal antibodies against HPV, oncoprotein p53, anti-apoptotic protein Bcl-2, and proliferation marker PCNA. The percentage of positive cells was determined as strongly, moderately or weakly positive. Results. Premalignant and malignant epidermal lesions failed to show specific clinical features suggesting that they are unlikely to be reliably distinguished on clinical signs alone and morphological examination seems to be mandatory. Immunohistochemistry revealed that HPV infection was 1.5 times more frequent in Bowen`s disease, than in invasive skin tumors. HPV+/p53+ types of Bowen`s disease showed higher PCNA expression in comparison to HPV-/p53+ types. Bcl-2 expression was associated with HPV-negativity. In HPV+ and

HPV- types of basal cell carcinoma and squamous cell carcinoma, the incidence of p53 and PCNA expression was similar. However, the strong expression of both markers (p53, PCNA) was related to the lower grade of squamous cell carcinoma. Conclusion. HPV infection appears to play a role in development of Bowen`s disease and malignant epidermal tumors. Independently of p53 expression, HPV-positive cases of Bowen`s disease exhibit higher proliferation activity, whereas HPV-negative cases are also Bcl-2-positive. Independently of the presence of HPV infection, basal cell carcinomas and squamous cell carcinomas show similar incidence of p53 expression and proliferation activity, and lower grade squamous cell carcinomas are associated with stronger expression of p53 and PCNA. PP3-195 EVALUATION OF CORRELATION BETWEEN CLINICAL AND PARACLINICAL DIAGNOSIS IN FEMALE PATIENTS WITH DIFFUSE NONCICATRICIAL ALOPECIA Pouran Layegh, Fakhrozaman Pezeshkpoor, Maryam Jannati Pour Qaem Hospital, Mashad University of Medical Sciences, Iran Background: Alopecia in all its patterns is a common symptom in patients who referred to dermatology clinics. It may cause anxiety and stress in these cases. There are four main diagnosis for a woman presenting with the chief complaint of diffuse, nonscaring hair loss: 1) Female pattern hair loss (androgenetic alopecia) 2) Acute and chronic telogen effluvium 3) Diffuse alopecia areata 4) Loose anagen hair syndrome. Management of hair diseases is very difficult, so we aimed to evaluate the correlation between paraclinical diagnosis (hormone studies, scalp skin biopsy/ histopathology) and clinical diagnosis (history and physical examination) in female patients with diffuse noncicatricial alopecia refer to clinic of dermatology of Ghaem Hospital. Method: in this prospective study, we filled out the questionnaires for 41 cases and their history, physical examination and hormonal and histopathological studies were recorded and final diagnosis was assessed. Then the correlation between clinical and paraclinical diagnosis were evaluated with Chi square test. Results: Most of patients had androgenetic alopecia. Clinical and paraclinical diagnoses were the same in 94.1 % of these cases. Diffuse alopecia areata was diagnosed by pathologic studies in all cases. Clinical diagnosis and hormone studies were the same in most cases of androgenetic alopecia and telogen effluvium. Conclusion: Pertinent history, physical examination and biopsy are suggested in all patients with diffuse alopecia. Hormone studies are not needed unless they have been other signs of hyperandrogenemia. PP3-196 EXPRESSION AND LOCALIZATION OF THYMIDINE PHOSPHORYLASE/ PLATELET-DERIVED ENDOTHELIAL CELL GROWTH FACTOR IN CUTANEUS MALIGNANT MELANOMAS Gultekin Kaner1, Fatma Kaya Dagistanli2, Selma Yilmazer2 1 Istanbul University, Cerrahpasa Medical Faculty, Department of Pathology, Turkey 2 Istanbul University, Cerrahpasa Medical Faculty, Department of Medical Biology, Turkey Backgraund: Angiogenesis is a significant prognostic factor in melanoma, but the angiogenetic factors controlling the neovascularization are not well defined. Platelet-Derived Endothelial Cell Growth Factor (PD-ECGF), also known as thymidine phosphorylase (TP) stimulates endothelial cell mitogenesis and chemotaxis in vitro and is strongly angiogenic in vivo. TP is overexpressed in various tumors and plays an important role in angiogenesis, tumor growth, invasion and metastasis. The aim of the study was to examine the expression

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of TP in melanomas. Method: In this study the expression of thymidine phosphorylase was examined immunohistochemically in 14 cases with melanomas. Result: TP was strongly expressed not only in macrophages in and around tumors but also in fibroblasts and keratinocytes. Immunoreactivity of TP was rarely seen in melanoma cells. TP positive reaction was also observed in ecrine gland epithelium. Conclusion: These results suggest that TP expressing infiltrative macrophages and stromal cells in and around tumors may contribute to angiogenesis and involved in the growth and metastasis of malignant melanomas. PP3-197 L1 EXPRESSION IS NOT RELATED WITH SENTINEL LYMPH NODES METASTASIS IN MELANOMA PATIENTS Lorenzo Memeo1, Raffaella Giuffrida1, Tiziana Perin2, Adriana Scogna2, Vincenzo Canzonieri2 1 Department of Experimental Oncology, Mediterranean Institute of Oncology, Catania, Italy 2 Division of Pathology, CRO- Aviano National Cancer Institute, IRCCS, Aviano, Italy Background: Recently, the expression of the cell adhesion molecule L1 has been associated with the metastatic phenotype in both human and murine model of malignant melanoma. Thies et al showed a positive association between L1 expression and metastasis in a 10-year retrospective study. L1 is a transmembrane protein and consists of an extracellular region, containing six immunglobulin-like domains followed by five fibronectin III-like repeats, a transmembrane domain and a short, phylogenetically-conserved cytoplasmic domain. Although originally described as a neuronal cell adhesion molecule, L1 expression is not restricted to nervous tissues, but has also been detected in lymphocytes, granulocytes, in epithelial cells of the intestinal and urogenital tract, and in the epidermis. In addition to expression in normal tissues, L1 has also been detected in several highly malignant tumours, including osteogenic sarcoma, squamous cell carcinoma of the lung, rhabdomyosarcoma and retinoblastoma. Therefore we decided to study L1 expression by immunohistochemistry in 52 non-consecutive patients with cutaneous malignant melanoma which underwent sentinel lymph node biopsy (SNB). Methods: L1 immunostaining was evaluated in melanoma cells by a semiquantitative scoring system. The cutoff for the overexpression was set at 20% of positive cells, irrespective of staining intensity. Results: Breslow Score Index (BSI) in all cases ranged from 0.44 mm to 8 mm. Out of the 52 melanoma patients, 23 ( 44%) had lymph nodes metastasis and 17 of them had the Breslow Index Score (BSI) available, ranging from 0.9 mm to 8 mm (12 cases with BSI >1,5mm). The 29 melanoma without lymph nodes metastasis had a BSI ranging from 0.44 to 4.85 mm (16 cases with BSI >1.5 mm). L1 overexpression in primary tumors was found in 10/23 (43%) cases with sentinel lymph node metastasis, and 16/29 (55%) cases with no evidence of sentinel lymph node metastasis When L1 expression was evaluated in combination with a BSI < 1.5 mm, it was found overexpressed in 3/5 cases with lymph node metastasis and in 4/13 without lymph node metastasis. When cases were stratified for a BSI > 1.5 mm, L1 overexpression was found in 6/12 cases with lymph node metastasis, and in 12/16 without lymph node metastasis. Conclusion: Our data suggest that, in the melanoma settings, L1 plays an important role in predicting long term survival but is not statistically significant in predicting sentinel lymph node metastasis. PP3-198 LYMPHOEPITHELIOMA LIKE CARCINOMA OF THE SKIN FOLLOWING 5-FLUOROURACIL TREATMENT Huseyin Kemal Turkoz, Deniz Ozcan Okmeydani Training and Research Hospital, Department of Pathology, Turkey

Background: Primary lymphoepithelioma like carcinoma (LELC) of the skin is a cutaneous malignancy with microscopic smilarities to undifferentiated nasopharyngeal carcinoma. It is a very rare neoplasm and just over 30 cases have been reported in the medical literature to date. Relation with previous actinic keratosis was not observed in the reported cases. Methods: We report a case of LELC on the right cheek of a 79 years old female who was treated with 5-fluorouracil for breast carcinoma five years before the onset of LELC. 5-fluorouracil is known to increase sensitivity to sun light. Patient had multiple actinic keratosis on her face three years after the 5-Fluorouracil therapy. All but one of the lesions regressed following topical diclofenac therapy and avoidance of direct sun light. The only remaining lesion was surgically removed with safe margins. Histologically, there was no connection between the tumor and the epidermis. There was no displasia in the neighbouring skin. The entire dermis and superficial subcutaneous tissue were occupied by atypical epithelial cell nests with a syncytial pattern of growth. Tumor cells were positive for cytokeratine and S-100 positive clear cells resembling Langerhans cells were scattered between tumor nests. Conclusions: None of the previously reported cases have been announced to be associated with neighbouring dysplasia or previous actinic keratosis in the vicinity of tumor location. This is the first LELC case which can be connected with sun light exposure and actinic keratosis as the precursor lesion of skin carcinoma of squamous type. PP3-199 A NEW CONCEPT OF MELANOCYTIC NEOPLASIA PATHOGENESIS BASED ON THE PHENOTYPE OF COMMON ACQUIRED NEVI Anna Batistatou1, Aikaterini Zioga1, John Panelos1, Daniela Massi2, Sevasti Kamina1, Konstantinos Charalabopoulos3, Niki Agnantis1 1 Department of Pathology, University of Ioannina Medical School, Ioannina, Greece 2 Department of Human Pathology and Oncology, University of Florence, Florence, Italy 3 Department of Physiology, Clinical Unit, University of Ioannina Medical School, Ioannina, Greece Background: To date, there are no known molecular hallmarks for nevus development but much progress has been made on the classification of melanocytic nevi. Common acquired nevi are classified into junctional, compound and dermal, based on the topography of nevus cells. The aim of this study was to examine whether common acquired nevi excised from the same individual share similarities in their microscopic appearance and in their molecular profile. Method: We retrieved from our files all the reports on excised nevi during the period 2/2005-2/2006. From these we selected only those that were more than one per patient and were diagnosed as common acquired nevi, compound or dermal. In total, our material consisted of 84 nevi from 31 individuals (Ƃ:ƃ=25:6, mean age: 31 years). Three patients had four excised nevi, 16 had three and 12 had two excised nevi. The histological slides were reviewed and all phenotypical characteristics were recorded. Furthermore, the immunohistochemical expression of E-cadherin (E-cad) was evaluated, using the monoclonal antibody (CM170B, Biocare Medical, Menarini Hellas). Results: Multiple nevi from the same individual share similar morphology, including secondary changes, some of which are not common. Such features include exophytic, papillomatous, acrochordon-like or endophytic architecture, cytologic characteristics, such as nuclear inclusions, predominant type A, B or C cells, multinuclear nevus cells, fatty metaplasia and “active” nevus cells. Regarding the immunohistochemical expression of E-cadherin in these nevi similar changes in expression in nevi of each individual were noted. Conclusion: On the basis of these observations, we hypothesize that melanocytes of the whole body, although

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dispersed in the entire epidermis, are all genetically similar in the same individual and changes predisposing to neoplasia are possibly a global melanocytic event characteristic for each person. PP3-200 LOSS OF BMI-1 EXPRESSION IS ASSOCIATED WITH THE CLINICAL PROGRESS OF MALIGNANT MELANOMA Ingeborg M Bachmann1, Hanne E Eknes2, Arie P Otte3, Lars A Akslen2 1 The Gade Institute, Section for Pathology, University of Bergen, and Department of Dermatology, Haukeland University Hospital, Bergen, Norway 2 The Gade Institute, Section for Pathology, University of Bergen, Haukeland University Hospital, Bergen, Norway 3 Department of Biochemistry, Swammerdam Institute for Life Sciences, University of Amsterdam, Amsterdam, The Netherlands Background The incidence of cutaneous melanoma is increasing, and there is a continuous search for improved prognostic markers and novel therapeutic approaches. BMI-1, a candidate stem cell marker is part of the Polycomb group of genes (PcG), which are important in embryonic gene regulation and tumor development. It has been suggested that BMI-1 protein is important in cell cycle regulation since both p16/INK4a and p14/ARF are downstream BMI-1 targets. Further, BMI-1 has been linked to the development of human malignancies of the haematopoietic and lymphatic systems, but its role in melanocytic tumors of the skin is unknown. Method BMI-1 staining in tumor cells was studied by immunohistochemistry of TMA sections made from 178 benign and malignant melanocytic lesions. We used two different monoclonal antibodies (clone F6, Upstate, Lake Placid, NY and the non-commercial clone 6C9). The arrays were scored blindly and there was good correlation between the two investigated antibodies (Spearman's rho correlation coefficient 0.62, p<0.0001). The staining was recorded using a semi-quantitative and subjective grading, considering both the intensity of staining and the proportion of tumor cells showing unequivocal positive reaction. A staining index (SI) was calculated as a product of staining intensity (0-3) and area of positive tumor cell nuclei (1<10 %, 2=10 %-50 %, 3>50 %). Results The immunohistochemical staining of BMI-1 showed a predominately nuclear staining pattern. BMI-1 expression was found to be significantly weaker in melanomas than in benign nevi. In established melanomas, loss of BMI-1 expression was associated with features of aggressive tumors, like increased tumor cell proliferation, presence of necrosis, and increased expression of both N-cadherin and ȕ3-integrin. Low levels of BMI-1 expression were significantly associated with decreased patient survival. Conclusions In conclusion, we demonstrate for the first time that loss of the candidate stem cell marker BMI-1 expression is associated with increased proliferative capacity, features of aggressive melanoma subgroups and reduced patient survival in established vertical growth phase melanomas. We also demonstrate that expression of the BMI-1 protein is reduced in melanomas when compared with benign nevi. PP3-201 A ZONAL COMPARISON OF MIB-1, CYCLIN D1, ESTROGEN AND PROGESTERONE RECEPTOR EXPRESSION IN NEVI FROM WOMEN IN PREGNANCY, PUBERTY OR NONE OF THE TWO Anna Batistatou, Aikaterini Zioga, John Panelos, Michael Doukas, Nafsika Simou, Niki J. Agnantis Department of Pathology, University of Ioannina Medical School, Ioannina, Greece

Background: Pregnancy and puberty influence the clinical appearance of common acquired melanocytic nevi, however the histologic changes are controversial and the possible factors implicated are still under investigation. The aim of the present study is to investigate the possible differences in MIB-1 immunoreactivity (a cycle-associated nuclear antigen), in expression of cyclin D1 (a nuclear protein synthesized in early G1) and in expression of estrogen (ER) and progesterone receptors (PR) in common nevi from women in pregnancy, puberty, or none of the two. Method: Our material consisted of 43 common acquired compound or dermal nevi: 11 from pregnant women, 20 from age-matched women that were not pregnant and did not take contraceptive pills, and 12 from girls in puberty. For immunohistochemical staining the antibodies against MIB-1 (DAKO), Cyclin-D1 (Diagnostic Biosystem, Menarini Hellas), ER (Novocastra) and PR (Novocastra), and the EnVision system (DAKO) were used. Positive and negative nevus nuclei were counted in at least six randomly selected fields (X400) in each of the upper (superficial zone), middle (middle zone) and lower (deep zone) third of the dermal part of the nevus. Results: The nevi from pregnant women, exhibited a statistically significant increase in MIB-1 immunoreactivity, more pronounced in the deep zone, compared to controls. In nevi from girls in puberty the counts were in between the other two categories. Cyclin D1 expression was weak and detected in <5% of nevus cells in nevi from all categories, but was more pronounced in nevi from pregnant women. ER expression was rarely detected, only in the pregnant women nevi. The majority of nevi in all three categories expressed PR. Conclusion: Nevi from pregnant women differ from those of non-pregnant women regarding MIB-1 immunoreactivity, cyclin D1 and ER expression. In girls in puberty, nevi exhibit an increase in the MIB-1 index. PP3-202 THE IMPORTANCE OF TENASCIN AND UBIQUITIN IN ESTIMATION OF WOUND AGE Ekin Ozgur Aktas1, Safiye Aktas2, Hulya Guler1, Huseyin Karaali1 1 Ege University Faculty of Medicine Department of Forensic Sciences, Izmir, Turkey 2 Dr Behcet Uz Children Research Hospital, Izmir, Turkey Background: Human wound age determination is of prime importance in forensic sciences. The different reactions of tissues to different natures of wounds require modern diagnostic methods to determine wound age. Ubiquitin is a small protein required for ATP-dependent, nonlysosomal intracellular protein degradation, which eliminates most intracellular defective proteins with a rapid turnover. It is expressed in nuclei of neutrophil leucocytes, macrophages and fibroblasts in wound area. Tenascin is an extracellular matrix glycoprotein. It is expression is restricted in adult tissues although it is widely distributed in embryonic tissues. The aim of this study is to determine the importance of ubiquitin and tenascin in wound age. Method: This study included 170 wound biopsies prospectively obtained from 89 forensic autopsy cases with known wound age. Ubiquitin and tenascin were immunohistochemically applied on formalin-fixed paraffin-embedded tissues. Pearson correlation analysis was performed. Results: Seventy four cases (83.15%) were male, while 15 cases (16.85%) were female. The mean age was 39.44 years. Among 170 wound samples 74 (43.5%) were gunshot wound, 20 (11.8%) were blunt injury and 76 (44.7%) were sharp weapon injury or surgical excision. Tenascin was negative in 123 cases (72.4%) in all series; it was negative in 98.3% in cases with wound age under 24 hours. It was positive in 91.8% in cases with wound age over 24 hours. Mean number of cells that express ubiquitin was 10.56%, while it was 4.25% in cases less than 24 hour wound age and, it was 26.14% in cases over 24 hours wound age. In correlation analysis both tenascin and ubiquitin were positively correlated with wound age. But in

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cases with wound age over 40 days tenascin becomes negative and ubiquitin is still expressed in fibroblasts. Conclusion: We conclude that tenascin and ubiquitin together is useful in determining wound age semi quantitatively. PP3-203 MERKEL CELL CARCINOMA – TWO CASE REPORTS Anastasia Nikolaidou1, Rodi Kotakidou2, Christos Tsalikidis3, Athena Kriaka2 1 Department of Pathology, General Hospital of Kilkis, Greece 2 Department of Pathology, General Hospital “G. Gennimatas”, Thessaloniki, Greece 3 2nd Department of Surgery, University Hospital of Alexandroupoli, Greece BACKROUND: Merkel cell carcinoma (MCC) is a distinctive cutaneous malignancy also known as neuroendocrine carcinoma of the skin. It is a rare disease in which malignant cells are located on or gust beneath the skin and in hair follicles. MCC has a malignant potential and thus prompt aggressive treatment is indicated. METHODS & RESULTS: We report two cases, a 28 years old male with a subcutaneous tumor located in the upper left arm measuring 2 cm and a 66 years old female with a subcutaneous tumor located in the buttock measuring 4 cm. The tumors were completely excised and submitted for histopathological examination. The latter showed dediffentiated malignant neoplasms consisting of small and medium sized cells with scanty cytoplasm, oval, hyperchromatic nuclei and numerous mitotic figures. The tumor cells were arranged in a diffused pattern or formed tiny nodules. The tumor extended deeply into the subcutaneous fat. The diagnosis of MCC was confirmed by histological and immunohistochemical analysis. The tumor cells stained positively for neurospecific- enolase, synaptophysin , chromogranin A, cytokeratin 20 and cytokeratin AE1/AE3. CONCLUSION: The differential diagnosis of MCC contains a vast variety of entities among which primary or metastatic malignant tumors are included. It is therefore important to combine physical examination detailed historical data and thorough histological immunohistochemical, ultrastructural even cytogenetic study in order to establish a prompt diagnosis PP3-204 UNCOMMON METASTASES OF MELANOMA WITHOUT KNOWN PRIMARY LESIONS Anastasia Nikolaidou1, Elisavet Pazarli1, Michael Katsamakas2, Christos Tsalikidis3, Andreas Tsalikidis2, Anastasia Kiziridou4 1 Department of Pathology, General Hospital of Kilkis, Greece 2 Department of Surgery, General Hospital of Kilkis, Greece 3 2nd Department of Surgery, University Hospital of Alexandroupoli, Greece 4 Department of Pathology, ‘’Theagenion’’ Anticancer Institute of Thessaloniki, Greece BACKGROUND: Clinically and histologically melanomas exhibit variant patterns of presentation. Melanoma is one of the malignant neoplasms that can regress, partially or completelly. The incidence of complete regression is 2,4 - 8,7 %. This means that many tumors considered to be primary could be metastases after the true primary lesion has regressed. METHOD AND RESULTS: We report four cases of male patients, 69 – 81 years old with unusual metastases, two with gastric melanoma, one at the inguinocrural area and one of the oral mucosa. None of them had a history of primary dermal melanoma. Two of the patients presented with symptoms of the gastrointestinal tract, one patient for treating incarcerated inguinal hernia and the fourth for ulcerative lesion of the oral cavity. The first two patients underwent gastroscopy and a biopsy was taken from an ulcerative neoplastic lesion of the stomach, the third one had a mass the size of tangerine surgically excised from his inguinocrural area and

the fourth had un ulcerative lesion also excised. According to the morphological and immunohistochemical findings (positivity for melanoma antigen, HMB-45, S-100 protein and vimentin and negativity for CEA, EMA, cytokeratin AE1 & AE3 and low molecular weight keratin) we diagnosed malignant melanoma in all cases. CONCLUSION: The cases we present are rare, especially in a general hospital. If there is no history of a prior melanoma, problems as far as the identification of the neoplasm can be set. Independently of the primary site or secondary tumor, we must keep in mind that melanomas must be included in differential diagnosis of a neoplasm with poor differentiation. PP3-205 MERKEL CELL CARCINOMA IN INGUINAL LYMPH NODE IN THE ABSENCE OF A PRIMARY SKIN TUMOR - A CASE REPORT Persefoni Xirou1, Ioannis Dimitriadis1, Kaliopi Gianna1, Doxakis Anestakis1, Barbara Christoforidou1, Konstantinos Setzis2, Frideriki Patakiouta1 1 Department of Pathology, “Theagenion” Cancer Hospital, Thessaloniki, Greece 2 Department of Surgery, “Theagenion” Cancer Hospital, Thessaloniki, Greece Introduction: Merkel cell carcinoma (MCC) is an aggressive primary neuroendocrine neoplasm of the skin, probably originating from precursor cells which give rise to keratinocytes and Merkel cells. It occurs mainly in elderly individuals, slightly more commonly in women, the head, neck and extremities being the most common location. Purpose: We report a rare case of MCC confined within an inguinal lymph node at the time of presentation. Case report: A 56 year old man presented with a large deep-seated mass in the right inguinal region, measuring 6 cm in maximum diameter, which was surgically resected. Histological examination revealed a lymph node extensively involved by malignant neoplasm with histological and immunohistochemical features of Merkel cell carcinoma. After thorough investigation of the patient, no primary skin tumour was found. Discussion: Merkel cell carcinoma is a highly malignant primary tumour of the skin, known for its propensity to metastasize early. MCCs within lymph nodes in the absence of a primary tumour have been sporadically reported. These rare malignant neoplasms might repesent metastases from an occult or totally regressed primary carcinoma. Other proposed explanations for their occurrence are malignant transformation of preexisting intranodal epithelial inclusions or anomalous carcinomatous differentiation of stem cells of the lymphoreticular system. PP3-206 MALIGNANT GLOMUS TUMOR ARISING FROM A BENIGN GLOMUS TUMOR: A CASE REPORT Roberto Salmaso1, Marina Gardiman1, Donatella Mannicci2, Amihood Cohen3, Ambrogio Fassina4 1 Department of Oncological and Surgical Sciences Section of Pathology Padua University, Italy 2 Department of Gynecological Science, Padua University, Italy 3 Department of Plastic Surgery, Padua University, Italy 4 Department of Oncological and Surgical Sciences, Section of Pathology, Unit of Cytopathology Padua University, Italy Glomus Tumours (GT) are benign tumours originatin from modified smooth muscle cells of the normal glomus body. GT are solitary, small size, painful lesions usually localized in the upper extremities, with predilection of subungueal regions. Rarely, GT may display unusual features as large size, infiltrative growth, mitotic activity, nuclear pleomorphism and necrosis.A 46 year old man presented with a painful, rapidly enlarging, dermalipodermal nodule noted three months before on the medial face of the right leg. The lesion was surgically removed without problems and grossly was cm 2,2 in diameter, solid, non necrotic

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and well circumscribed from the surrounding adipose tissues. Microscopically, the lesion appeared multinodular, circumscribed by a fibrous capsule, and composed by a population of round and fusiform cells with round nuclei and eosinophilic cytoplasms. The neoplastic glomic cells were organized in monotonous nests and sheets and formed collars around many blood vessels with prominent endothelial cells. Near the margins of the surgical excision, there were two accessory highly cellular nodules, with marked nuclear atypia, elevated mitotic index (> 30 mitoses / 50 HPF) with atypical mitotic figures, and with rare vascular channels.Immunohistochemical staining for alpha-smooth muscle actin and type IV collagen were suggestive for the glomic origin of the lesion, while MIB-1 reactivity demonstrated a strong difference between the two accessory nodules ( > 17%) and the larger neoplastic nodule ( < 2%). We concluded for a final diagnosis of malignant glomus tumour (MGT) arising in a benign glomus tumour. The differential diagnosis of MGT is broad. In the skin it can be confused with skin adnexal neoplasms including nodular hidradenoma, hidradenocarcinoma and eccrine spiradenoma. Demonstration of S-100 protein expression and HMB-45 positivity permits the distinction of malignant melanoma and clear cell sarcoma from MGT. Also small round blue cells tumour can be confused with MGT (cytokeratin 20 negative) including Merkel cell carcinoma (cytokeratin 20 positive), Ewing sarcoma and PNET (both muscle actins negative). In the deep soft tissues, MGT (CD 34 negative) should be distinguished from hemangiopericytoma (CD 34 positive), leyomiosarcoma and rhabdomyosarcoma. MGT is a low-grade malignant tumour, locally aggressive but rarely metastatizing, although more than 25% of the reported cases developped metastasis. A wide local excision with histopathologically free margins and close follow up is considered the treatment of choice.

PP3-208 SILVER STAINED ORGANIZER REGIONS AND IMMUNOGLOBULINS IN CUTANEOUS KERATOACANTHOMAS AND SQUAMOUS CELL CARCINOMAS Kiriaki Aroni, Aikaterini Mastoraki Department of Dermatopathology, Kapodistrian University of Athens, Greece

PP3-207 A DERMATOFIBROSARCOMA PROTUBERANS (DFSP) OCCURRING IN A SITE OF MULTIPLE CORTICOID INJECTIONS Mameri Saâdia, Bouzid-Bendisari Kheira CHU Béni-Messous, Algeria

PP3-209 ECCRINE SPIRADENOMA MIMICKING THYMOMA Stephanie Vgenopoulou1, Georgia Levidou2, Athina Androulaki1, Grigorios Kouraklis3 1 First Department of Pathology, University of Athens, Laiko Hospital, Greece 2 National and Kapodistrian University of Athens, School of Medicine, First Department of Pathology, Greece 3 Second Department of Surgery, University of Athens Medical School, Laiko Hospital, Greece

Background: DFSP is a mesenchymal sub-cutaneous tumor characterized by a strong tendency to recur, rarely metastasizing, which grows very slowly. In some instances, inflammatory conditions like trauma, sites of injections, vaccinal scars, burns have preceded the apparition of the DFSP. Method: We report one case concerning a woman of 53 years old .She has been affected 8 years ago by a DFSP on the left shoulder. This one occurred on the site of multiple corticoïd injections for a severe rheumatoid polyarthritis. The clinical exam reveals a round mass, firm covered by a very thin skin, badly limited in the hypodermis with a pink section. Many tumor specimens are realized and stained by hematoxylin-eosine. Results: The microscopic analysis shows a dermal tumor with a Grenz zone.It is composed of spindle cells arranged in a classical storiform pattern (cartwheel pattern).An immunostain for CD34 gives a diffusely and intensively positivity confirming the diagnosis of DFSP. Conclusion: Some patients relate their DFSP to a prior trauma. Only few authors report factors or conditions which precede the apparition of the lesion while others consider these factors just like a coincidence. This aspect must be known in order to control every imperfectly cicatrisation after a vaccination or repeated injections. This imperfectly scar must be analysed because the recommended surgical resection of a DFSP is a wide deeply excision with margins as large as possible.

Background: The aim of the study was to investigate the epidermal cells biological activity in Keratoacanthomas (KAs) and Squamous Cell Carcinomas (SCCs) by counting the number of silver stained Nucleolar Organizer Regions (AgNORs), to estimate the quantity of Ig-producing cells and the inflammatory cellular infiltrate (ICI) in these entities and to achieve a comparative evaluation of them. Method: Thirty KAs (10 in growth stage, 10 in mature stage and 10 in involution stage) and 28 SCCs (9 well differentiated-G1, 7 moderately differentiatedG2, 5 poorly differentiated-G3 and 7 pseudoadenoid) were investigated. Results: The KAs examined had a mean number of 1.727 AgNORs (S.D. 0.232) and IgG predominated in most cases. IgG and IgE increase at the involution stage, IgA remains at almost the same level in the three stages and IgM decreases during the maturity stage. The SCCs examined had a mean number of 2.105 AgNORs (S.D. 0.446). IgG predominated and gradually increases in proportion to the degree of malignancy. Conclusion: There is a significant difference in the number of AgNORs among the three stages of KA with an increase in the growth and involution stage and a drop during the maturity stage. The proportion of Ig subclasses was different in the three tumor stages in contrast to the cellular infiltrate. In SCCs, the number of AgNORs and the percentage of Igs and ICI increased gradually in proportion to the degree of malignancy.

Eccrine spiradenomas are well demarcated, lobular, often painful adenomas that can be encountered almost anywhere on the body. Occasionally, eccrine spiradenomas show a heavy lymphocytic infiltrate with a resulting appearance reminiscent of thymoma. Here we present such a case. Our case was investigated immunohistochemically with a wide range of antibodies, including lymphocytic and epithelial markers. A comparative study of histological and immunohistochemical characteristics of eccrine spiradenomas in contrast to thymomas was carried out. After a comprehensive study of both morphological and immunohistochemical parameters, we reached a diagnosis of eccrine spiradenoma. The lymphocytic markers CD1a and CD99 which are markers of an immature T-cell phenotype, were found to be negative in our case, verifying the diagnosis of spiradenoma. Despite overlapping histomorphological features of eccrine spiradenoma and thymoma, the immunophenotype of the lymphocytes is helpful in the differential diagnosis of the two entities. In thymomas, the T-cell population has an immature immunophenotype and shows positive staining for CD1a and CD99, while in spiradenomas, the T-cell population has a mature immunophenotype and is hence negative for the above mentioned markers.

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PP3-210 THE SIGNIFICIANCE OF HISTOLOGICAL PATERNS IN POSITIVE PPD TEST SITE Aptullah Haholu1, Faruk Ciftci2, Ozlem Karabudak3, Ali Kutlu4, Bulent Bozkurt4, Huseyin Baloglu1 1 Gulhane Medical Academy Haydarpasa Training Hospital Deparment of Pathology, Turkey 2 Gulhane Medical Academy Haydarpasa Training Hospital Deparment of Chest Disease, Turkey 3 Gulhane Medical Academy Haydarpasa Training Hospital Deparment of Dermatology, Turkey 4 Gulhane Medical Academy Haydarpasa Training Hospital Deparment of Allergy, Turkey BACKGROUND: The histologic features of tuberculin skin test site is not uniform. It may be related to status of tuberculosis. The aim of this study was to compare the histological patterns of tuberculin skin test reaction of the patients with active and latent pulmonary tuberculosis and to investigate the histopathological differences between them. METHODS: Forty-eight PPD positive cases were chosen for the study among the patients admitted to Gulhane Medical Academy, Haydarpasa Training Hospital, Department of Chest Disease in the year 2006. Thirty of the subjects had active tuberculosis. A 3 mm punch-biopsy specimen was taken from the central portion of the skin reaction after obtaining informed consent from these PPD positive subjects. They were processed in a usual way. Sections were stained with hematoxylin-eosin and examined under light microscope. The histological patterns of reactions were classified into three types: (a) Perivascular (PV) type, (b) Basal spongiotic dermatitis (BSD) type, (c) erythema multiforme (EM) type. The histological patterns in active tuberculosis and latent tuberculosis were comparatively evaluated by statistical analysis. RESULTS: There were 17 (56.7%) EM type, 9 (30%) BSD-type and 4 (%13.3) PVtype inflammatory reactions among 30 active tuberculosis patients. There were 2 (11.1%) EM-type, 7(38.8%) BSD-type and 9 (50%) PV-type inflammatory reactions among 18 latent tuberculosis patients. The EM type inflammations were frequently seen in active tuberculosis group than in latent tuberculosis (56.7% vs. 11.1%) (p<0.05). However, the PM type inflammations were less frequently seen in active tuberculosis group than in latent tuberculosis (%13.3 vs. 50%) (p<0.05). The BSD type inflammation frequencies between both groups were not significant. Interestingly, the bullae formations were only seen in 7 subjects with active tuberculosis. CONCLUSION: The histological pattern of PPD reaction sites may be an important sign reflecting the nature of the tuberculosis, which may be either latent or active. The bullae formation, another histological finding in our study, seems to be a specific sign for active pulmonary tuberculosis. Further detailed immunohistopathological studies of PPD biopsies including large number of cases might give important clues about tuberculosis immunology. PP3-211 CHARACTERISTICS OF PRIMARY CUTANEOUS BCELL LYMPHOMAS: CLINICOPATHOLOGIC FEATURES, IMMUNOPHENOTYPIC PROFILE AND PROGNOSTIC FACTORS Lebriz Uslu1, Cuyan Demirkesen1, Oya Oguz2, Nahide Onsun3, Nukhet Tuzuner1 1 Department of Pathology, Cerrahpasa Medical Faculty,Istanbul University, Istanbul, Turkey 2 Department of Dermatology, Cerrahpasa Medical Faculty,Istanbul University, Istanbul, Turkey 3 Department of Dermatology, Vakif Gureba Teaching Hospital, Istanbul, Turkey Background: In the recent WHO-EORTC classification for cutaneous lymphomas, primary cutaneous B-cell lymphoma

(PCBL) include cutaneous follicle center lymphoma (PCFCL), cutaneous marginal zone B-cell lymphoma (PCMZL), diffuse large B-cell lymphoma, “leg type” (PCLBCL-LT) and diffuse large B-cell lymphoma, “other type” (PCLBCL-O). Both PCFCL and PCMZL are indolent lymphomas with good prognosis, while PCLBCL has an intermediate-level prognosis. Method and results: In our study, 40 patients with PCBL were reviewed and reclassified according to the new classification scheme. All of the patients, diagnosed during 11-year period (1996-2007), were analyzed for clinicopathologic features, expression of several markers including CD20, Bcl-2, Bcl-6, MUM-1 and Ki-67, in situ hybridization for Epstein-Barr virus, and of Borrelia burgdorferi. The histologic features and the immunophenotypic profiles of each lymphoma group were stated. The patients were classified into the following categories: PCFCL, 16 patients, PCMZL, 16 patients, PCLBCL-LT, 6 patients and PCLBCL-O, 2 patients. The associations between histology, skin site, the number of lesions and survival were determined. Conclusion: The results underlined the clinical significance of the new classification, the importance of the localization and the number of lesions. PP3-212 PRIMARY CUTANEOUS CD30(+) LYMPHOPROLIFERATIVE DISORDERS: LYMPHOMATOID PAPULOSIS AND PRIMARY CUTANEOUS ANAPLASTIC LARGE CELL LYMPHOMA Erdem Caglar1, Cuyan Demirkesen1, Oya Oguz2, Nahide Onsun3, Nukhet Tuzuner1 1 Department of Pathology, Cerrahpasa Medical Faculty,Istanbul University, Istanbul, Turkey 2 Department of Dermatology, Cerrahpasa Medical Faculty,Istanbul University, Istanbul, Turkey 3 Department of Dermatology, Vakif Gureba Teaching Hospital, Istanbul, Turkey Background: Primary cutaneous CD30(+) lymphoproliferative disorders (LPD) are considered as the second most common group among primary cutaneous T-cell lymphomas (CTCL). Method and results: We reviewed 74 cases displaying atypical CD30(+) large cells in their biopsies. All of the patients, diagnosed during 12-year period (1995-2007), were analyzed for clinicopathologic features, expression of several markers including CD3, CD5, CD4, CD8, CD56, CD30, ALK, EMA, cytotoxic proteins such as granzyme B and in situ hybridization for Epstein-Barr virus (EBV). The patients were categorized as: Lymphomatoid papulosis (LyP), 34 patients, primary cutaneous anaplastic large cell lymphoma (ALCL), 16 cases, borderline CD30(+) LPD, 5 patients, ALCL, B-cell type, 1 patient, MF transformed into CD30(+) large cell lymphoma, 9 patients. One case of LyP and one case of CD30(+) borderline LPD suffered also from mycosis fungoides (MF). ALCL, B-cell type was associated with EBV. Other than these, there were 2 cases of scabies and 6 cases of arthropod bite, showing numerous CD30(+) large T-cells. Conclusion: The results underlined the importance of clinicopathologic correlation. Sometimes the course should be used as decisive criteria for the definite diagnosis. The prognosis of primary cutaneous CD30(+) LPD were excellent, however, MF, transformed into CD30(+) large cell lymphoma had an aggressive course.

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PP3-213 PRIMARY CUTANEOUS T-CELL LYMPHOMAS OTHER THAN MYCOSIS FUNGOIDES AND PRIMARY CUTANEOUS CD30 (+) LYMPHOPROLIFERATIVE DISORDERS Murat Ozbalak1, Cuyan Demirkesen1, Oya Oguz2, Nahide Onsun3, Nukhet Tuzuner1 1 Department of Pathology, Cerrahpasa Medical Faculty,Istanbul University, Istanbul, Turkey 2 Department of Dermatology, Cerrahpasa Medical Faculty, Istanbul University, Istanbul, Turkey 3 Department of Dermatology, Vakif Gureba Teaching Hospital, Istanbul, Turkey Background: Among primary cutaneous T-cell lymphoma (CTCL) Mycosis fungoides (MF), together with its variants and subtypes, is the most common type of CTCL. The second most common type is primary cutaneous CD30(+) lymphoproliferative disorders (LPD), accounting for approximately 30% of CTCL. In the recent WHO-EORTC classification for cutaneous lymphomas, the rest is classified as subcutaneous panniculitislike T-cell lymphoma, extranodal NK/T-cell lymphoma, nasal type, primary cutaneous aggressive epidermotropic CD8(+) Tcell lymphoma (provisional), cutaneous Ȗ/į T-cell lymphoma (provisional), primary cutaneous CD4(+) small/medium-sized pleomorphic T-cell lymphoma (provisional) and , primary cutaneous peripheral T-cell lymphoma, unspecified. Method and results: In our study, 17 patients with CTCL other than MFand primary cutaneous CD30(+) LPD, were reviewed and reclassified according to the new classification scheme. All of the patients, diagnosed during 10-year period (1996-2006), were analyzed for clinicopathologic features, expression of several markers including CD3, CD5, CD4, CD8, CD56, CD30, granzyme B and in situ hybridization for Epstein-Barr virus. The patients were classified into the following categories: Subcutaneous panniculitis-like T-cell lymphoma, 1 patient, extranodal NK/Tcell lymphoma, nasal type, 4 patients, primary cutaneous CD4(+) small/medium-sized pleomorphic T-cell lymphoma, 3 patients, primary cutaneous peripheral T-cell lymphoma, unspecified, 5 patients. There were 3 MF and 1 pagetoid reticulosis patients, who were transformed into CD30(-) large T-cell lymphoma. Two patients were diagnosed as CD4(+)/CD56(+) hematodermic neoplasm (blastic NK cell lymphoma). Conclusion: The clinical and histologic features, the immunophenotypic profiles and their association with prognosis were analyzed. PP3-214 CLEAR CELL HIDRADENOCARCINOMA ARISING IN A NEVUS SEBACEUS Roberto Salmaso1, Marina Gardiman1, Amihood Cohen2, Donatella Mannicci3, Luciano Giacomelli4, Ambrogio Fassina4 1 Department of Oncological and Surgical Sciences Section of Pathology, Padua University, Italy 2 Department of Plastic Surgery, Padua University, Italy 3 Department of Gynecological Science, University of Padua, Italy 4 Department of Oncological and Surgical Sciences, Section of Pathology, Unit of Cytopathology, Padua University, Italy The nevus sebaceus of Jadassohn (SNJ) is a hamartomatous proliferation of the skin and adnexa arising in the face and scalp of adults, with frequent association with several neoplasms, most commonly basal cell carcinoma, trichoblastoma, tichoepitelioma and syringocystadenoma papilliferum. We report the case of a 46 year-old female with hidro-adenocarcinoma demonstrating both apocrine and eccrine differentiation arising in a long standing nevus sebaceus on the scalp. Microscopically, the epidermis showed focal benign basaloid hyperplasia and irregular follicles. A small dermal collection of basaloid sebocytes was suggestive of a sebaceoma. The large part of the lesion was composed of

lobules of epithelium with focal cystic spaces. The cells showed high mitotic activity and nuclear pleomorphism, with cytoplasmic vacuoles containing abundant glycogen. Cells with eosinophilic cytoplasm were evident in areas adjacent to groups of basaloid cells with no vacuolation very similar to basal cell carcinoma differentiation. Ductal differentiation with well-formed papillae and focal cribriform structures were also present. Other portions demonstrated a high-grade neoplasm with prominent nuclear atypia and a solid pattern of growth resembling ductal breast carcinoma. EMA strongly stained tumor cells and highlighted the ductal origin of these structures. Less than 4% of cells stained for progesterone and estrogen receptors. Her2/neu reactivity was focally present showing 1+ membranous reactivity in 10% of cells. Anti-p63 labelled the basaloid cells surrounding the tumor lobules. A breast primary was ruled out by clinical and radiological examination. Other tumors showing conspicuous cytoplasmatic vacuolation enter the differential diagnosis, including clear cells squamous carcinoma, trichilemmal carcinoma and metastatic clear renal cell carcinoma, lung, liver and female genital tract. This report illustrates an extraordinary adnexal neoplasm with multiple lines of differentiation arising "de novo" in a SNJ. PP3-215 FOLLICULO-SEBACEOUS CYSTIC HAMARTOMA: REPORT OF 16 CASES Fernando Terrasa1, Alexandra Gené2, David R Berk3, Carlos Saus2, Rosa Taberner1, Daniel Santa Cruz4 1 Hospital Son Llatzer, Mallorca, Spain 2 Hospital Son Dureta, Mallorca, Spain 3 Washington University School of Medicine, St. Louis,MO, USA 4 Cutaneous Pathology, St. Louis, MO, USA BACKGROUND Folliculo-sebaceous cystic hamartoma (FSCH), described by Kimura et al. in 1991, is an uncommon malformation composed of epithelial and nonepithelial elements that grow into an infundibular cyst-like structure, with sebaceous glands and ducts radiating from it, and surrounded by a fibrous stroma with variable amounts of vascular or adipose tissue. Some authors have reported neural elements or myxoid change in the stroma. METHODS We performed a retrospective search from 3 institutions between 1999 and 2006 and identified 16 cases. Specimens from all 16 cases had been fixed in formalin and routinely processed. Five m sections were obtained and stained with hematoxylin and eosin. Alcian blue stain was also done in some cases to highlight myxoid change in the stroma. RESULTS We report 16 cases of FSCH including 11 women and 5 men, ranging from 24 to 87 years-old. Six lesions occurred on the nose, 5 on the trunk, 3 on the scalp, 1 on the cheek and 1 on the labia minora. Epidermal induction, similar to that encountered in dermatofibroma, was present in 10 cases. Reactive melanocytic hyperplasia was present in 5 cases, including 1 which was prominent, similar to junctional nevus. Myxoid stromal change occurred in 4 cases. Hair shafts in the infundibulum were present in 4 cases. One case that presented as several papules around a scar was a recurrence of a hamartoma removed 7 years earlier. CONCLUSIONS FSCH is a rare benign skin lesion, probably underdiagnosed due to its wide range of histological appearances. We report here 16 cases of FSCH, the largest series of this lesion to date. Some rarely described features such as myxoid stromal change, presence of hair shafts in the infundibulum, epidermal induction and melanocytic hyperplasia appear to be fairly common. We also report a previously undescribed mucosal location (labia minora), and the first local recurrence. Histological recognition of FSCH is crucial to distinguish it from other mucinous or hamartomatous lesions.

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PP3-216 ROLE OF MISMATCH REPAIR PROTEINS AND TELOMERASE IN CELL KINETICS OF SUPERFICIAL SQUAMOUS CELL CARCINOMA OF THE SKIN Alfredo Blanes1, Inmaculada Ruiz1, Juan José Sánchez-Carrillo1, Blanca Cabra De Luna1, Salvador J Diaz-Cano2 1 University Hospital of Malaga School of Medicine, Malaga, Spain 2 Department of Histopathology, King’s College Hospital, London, United Kingdom Background: The cell kinetic of superficial squamous cell carcinoma (SCC) has revealed controversial results due to the heterogeneity of lesions considered, making more difficult to assess key elements in squamous cell tumor progression. This study analyzes kinetic, cell survival and mismatch repair in a series of high-grade intraepithelial and microinvasive SCC. Design: We selected bowenoid actinic keratosis (HG-AK, 22 cases), SCC in-situ (37 cases) and microinvasive SCC (<3mm depth, 36 cases) that have appropriate archival material. Representative samples were evaluated by standard immunohistochemistry for MCM2 (minichromosome maintenance-2), telomerase, mlh1, msh2 and TP53. The immunoexpression was assessed in the whole lesion and the positive cells as percentage of tumor cells. Appropriate controls were run in each sample. The results were statistically compared using analysis of variance and Student t-test, and considered significant if P<0.05. Results: MCM2 inmunoreactivity was observed in 9% of cellularity of AK-HG, 19.17% of SCC INSITU and 28.58% of SCC MICRO. TELOMERASE was expressed in 55.75% (AK-HG), 48.08% (SCC IN-SITU) and 72.39% (SCC MICRO). MLH1/MSH2: in 7.73% (AK-HG), 19.04% (SCC IN-SITU) and 20.15% (SCC MICRO) (NULL: 58% AK-HG, 12.7% SCC IN-SITU and 10.86% SCC MICRO) and finally, TP53 inmunoreactivity in 30.5% (AK-HG), 32.78% (SCC IN-SITU) and 36.63% (SCC MICRO). Both MCM2 and TP53 expression increased from HG-AK to microinvasive SCC, revealing no statistically significant differences. Telomerase showed significantly upregulated expression in microinvasive SCC when compared with intraepidermal lesions (P=0.0037) and mlh1 expression was significantly down-regulated in HG-AK (P=0.0022). Conclusions: 1) The invasion capacity in superficial SCC is marked by up-regulation of telomerase in lesions preferentially expressing abnormal TP53. 2) The differential growth pattern of intraepidermal SCC is kinetically related with down-regulation of mismatch repair proteins. PP3-217 A COLLISION TUMOR OF THE SKIN ASSOCIATING BASAL CELL CARCINOMA AND NODULAR MALIGNANT MELANOMA (MALIGNANT BASOMELANOCYTIC TUMOR) Alexandra Bastian1, Geanina Micu1, Razvan Andrei1, Luciana Nichita1, Carmen Ardeleanu2, Florica Staniceanu3, Virginia Chitu4 1 Colentina University Hospital, Department of Pathology, Bucharest, Romania 2 Victor Babes Institute, Bucharest, Romania 3 Colentina University Hospital, Bucharest, Romania 4 Colentina University Hospital, Department of Dermatology, Bucharest, Romania Background: The coexistence of two malignant tumors in the same histological specimen is rare. A collision tumor consisting in basal cell carcinoma and malignant melanoma (malignant basomelanocytic tumor) has been rarely reported. Most of these case reports presented in situ malignant melanoma component. Material and method: We report the case of an 86 years old woman with a whitish, ulcerated tumor of the left cheek, macroscopically measuring 3/1.5/0.8 cm. Results: Histological

examination revealed basal cell carcinoma solid type with adenoid areas that invades the reticular dermis admixed with ulcerated and amelanotic, nodular malignant melanoma, Clark level IV and Breslow index 5.9 mm (maximum thickness of the tumor). In some areas the distinction of the two lesions was difficult. Cords and islands of basal cell carcinoma invade malignant melanoma nodule with intimate association of these two cell populations. Both lesions have been closely attached to the epidermis, data supporting primary cutaneous origin of both of the tumoral proliferations. Immunohistochemical studies showed that the malignant melanoma component was positive for S100 protein, HMB45 and CD63 and negative for CK34betaE12 and BerEp4, while the basal cell carcinoma component of the tumor was negative for S100 protein, HMB45 and CD63 and positive for CK34betaE12 and BerEp4. Conclusion: This collision tumor could be interpreted as a tumor with bidirectional keratinocytic and melanocytic differentiation or a coincidence, an extremely rare event. PP3-218 PROGNOSTIC FACTORS FOR RECCURRENCES IN BASAL CELL CARCINOMA Alexandra Bastian1, Geanina Micu1, Irina Tudose1, Adina Ene1, Razvan Andrei1, Sabina Zurac1, Eliza Gramada1, Dragos Simeanu1, Florica Staniceanu1, Dorina Giurcaneanu2 1 Colentina University Hospital, Department of Pathology, Bucharest, Romania 2 Colentina University Hospital, Department of Dermatology, Bucharest, Romania Background: Basal cell carcinoma (BCC) is the most common cutaneous malignant tumor that occurs at any age with increased incidence after the 4-th decade. BCC rarely metastasize (reported incidence of 0.0028-0.5%), but it causes a considerable morbidity by invading and destroying the surrounding tissue and location especially involving the head and neck region. Method: We studied 366 cases diagnosed in our department with BCC in two consecutive years. Among these, 335 cases were primary BCC (group A) and 31 were recurrences of a previously resected BCC (group B). We analyzed clinical informations, location, histological types and the integrality of excision. Results: Insignificant differences were recorded in respect of sex or age between the two groups, but the location of the tumors was different between the two groups. All the group B cases occurred in the head area (nose 32.25%, zygomatic 16.12%, perioral 12.90%, temporal 6.45%, orbital area 6.45%, frontal 3.22%, laterocervical 3.22%) while group A included 206 cases (61.49%) in the head area (orbital area 21.94%, nose 12%, zygomatic 6.56%, temporal 6.26%, frontal 4.18%, perioral 1.19%, laterocervical 0.89%) and 129 cases (38.50%) with other locations. There was a preponderance of adenoid, keratotic and metatipic types in both group A and B. We noticed that the infiltrative type of BCC was quite rare: 23 cases in group A (5.67%) and 4 in group B (1.09%). Incomplete excision was found in 93 cases (27.76%) of the group A and 13 cases (41.93%) of the group B. Conclusion: The aim of entire excision of BCC is difficult to obtain because of both macroscopic indistinct margins of tumors and the need of maximal skin preservation that appear in specific areas as eyelid or nose. There is even more difficult to entirely remove the recurrent BCC because of the preexisting scars that interfere with complete excision. Despite our expectations, we found a low correlation between the infiltrative histological subtype of BCC and recurrence. The location and the completeness of the excision have a stronger influence on the recurrence rate than the histological type. The adequate excision of the primary tumor is the key of surgical treatment in BCC.

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Head and Neck Pathology PP3-219 LYMPHOEPITHELIAL CARCINOMA OF THE LARYNX IN A 60-YEAR-OLD TUNISIAN MAN Sarra Mestiri, Amel Trabelsi, InéS Kalamoun, Lilia Ben Yacoub-Abid, Thouraya Aachech, Badreddine Sriha, Moncef Mokni, Sadok Korbi Service de Pathologie, Hopital F. Hached, 4000 Sousse, Tunisie Background: Lymphoepithelial carcinoma of the larynx is extremely rare, representing less of 0,5 % of larynx carcinomas, it is uncommonly associated with Epstein Barr Virus. Method: We report a case of a 60 year-old Tunisian man, who presented with hoarseness, dyspnea. Laryngoscopy showed a tumor of the left larynx extending to the pharynx. Biopsy and the resection specimen were both submitted for histopathological study. In-situ hybridization was performed in the latter to detect the presence of Epstein-Barr virus . Results : Histopathological examination of the specimen biopsy showed an undifferentiated carcinoma with a prominent lympho-plasmocytic infiltrate. A pharyngolaryngectomy with left neck dissection was performed and the final diagnosis retained was a lymphoepithelial carcinoma non EBV induced. A post-operative radiotherapy was indicated. Clinical outcome was favourable one year later without any recurrence nor metastasis. Conclusion: Through this case and literature review, the diagnosis and therapeutic problems raised by this very rare tumor are discussed. PP3-220 CHRONIC SINO-NASAL PROGRESSIVE DESTRUCTIVE DISEASE IN COCAINE ABUSERS: A MORPHOLOGICAL STUDY AND ELABORATION OF A GRADING Barbara Paola Banzatti, Francesco Klinger, Valeria Bandi, Angelo Virgilio Pagliari, Marco Klinger Plastic Surgery Institute - University of Milan - Plastic Surgery Unit - Istituto Clinico Humanitas - IRCCS – Milano, Italy Background In the last decade cocaine consumption as a recreational drug by snorting or sniffing has worlwide increased. Cocaine can cause tissue damage virtually in every organ system. Cocaine's adverse effect on the nasal cavity was recognized and described first by Owens in 1910 and cause heterogeneous spectrum of lesions from ulceration and nasal septum perforation to idiopathic central facial destructive granuloma. Mehod A population of 16 cocaine abusers patients was carefully investigated by the standard clinical, laboratory, radiological methods and multiple biopsies of the nose. Results In all cases the erythrocyte sedimentation rate was elevated, but the complete blood count, antinuclear, antimithocondrial, anti-DNA, antineutrophil cytoplasmatic antibodies (c-Anca) and bacteriological cultures were normal. Computed tomography showed destruction of nasal septum in 11 cases, of inferior and middle turbinate in 3 cases, of medial wall of maxillary sinus and of hard palate in one patient. Biopsy specimens showed no evidence of malignancy, vasculitis or granulomatosis. In nasal mucosa microscopy found an acute, chronic and partly necrotising inflammation process. Interesting in this condition there are "onion skin" deposits of perivascular fibrosis with numerous eosiniphils. Plasma cells and lymphocytes mainly of T cell type were present in early stage. Conclusion Cocaine damage nose by a combination of intense vasoconstriction and irritation. The ischemia and irritation of the nasal mucosa lead initially to an acute inflammation clinically visualize as a dystrophic lesions (initial stage or grade 1 of cocaine lesion), then a chronic inflammation characterized by ulceration and crusting (intermediate stage or grade 2) and finally various and deeply necrosis process that cause a destruction of normal anatomic strctures such as septum, turbinate and palate (advance stage or grade 3). The altered muco-ciliary clearance and air flow together came up infection cause a continued expansion and progression of the clinico-pathologic process.

PP3-221 SURVIVIN AND INDUCIBLE NITRIC OXIDE SYNTHASE PRODUCTION DURING 4NQO-INDUCED RAT TONGUE CARCINOGENESIS: A POSSIBLE RELATIONSHIP Daniel Araki Ribeiro1, Darcio Kitakawa2, Maria Aparecida Custódio Domingues2, Luiz Antonio Guimaraes Cabral2, Mariangela Esther Alencar Marques2, Daisy Maria Favero Salvadori2 1 Federal University of Sao Paulo, UNIFESP, Brazil 2 Sao Paulo State University, UNESP, Brazil Background. This study was undertaken to investigate, by immunohistochemistry, the expression of survivin and inducible nitric oxide synthase during 4NQO-induced rat tongue carcinogenesis. Methods. Male Wistar rats were distributed into three groups of 10 animals each and treated with 50 ppm 4NQO solution through their drinking water for 4, 12, and 20 weeks. Ten animals were used as negative control. Results. Although no histopathological abnormalities were induced in the epithelium after 4 weeks of carcinogen exposure, survivin and iNOS were expresssed (p<0.05) in some cells of the ‘normal’ oral epithelium. In pre-neoplastic lesions at 12 weeks following carcinogen exposure, the levels of survivin and iNOS were increased (p<0.05) when compared to negative control, being the strongest effect observed to iNOS. In well-differentiated squamous cell carcinoma induced after 20 weeks of treatment with 4NQO, survivin and iNOS were expressed in some tumor cells. Lack of immunoreactivity for both markers was observed in the negative control group. Conclusion. Taken together, our results support the belief that expression of survivin and iNOS are early events during malignant transformation and conversion of the oral mucosa. PP3-222 PLACENTAL GLUTATHIONE S-TRANSFERASE CORRELATES WITH CELLULAR PROLIFERATION DURING RAT TONGUE CARCINOGENESIS INDUCED BY 4-NITROQUINOLINE 1-OXIDE Daniel Araki Ribeiro1, Renata Nunes Da Silva2, Daisy Maria Favero Salvadori2, Mariangela Esther Alencar Marques2 1 Federal University of Sao Paulo, UNIFESP , Brazil 2 Sao Paulo State University, UNESP , Brazil Background. Taking into consideration that glutatione Stransferase (GST) and cellular proliferation play a crucial role during carcinogenesis, the goal of this study was to investigate the expression of placental GST, called GST-P, and proliferating cellular nuclear antigen (PCNA) by means of immunohistochemistry during rat tongue carcinogenesis induced by 4-nitroquinoline 1-oxide (4NQO). This is a useful model for studying oral squamous cell carcinoma phase by phase. Method. Male Wistar rats were distributed into 3 groups of 10 animals each and treated with 50 ppm 4NQO solution by drinking water for 4, 12 or 20 weeks. Ten animals were used as negative control. Results. GST-P positive foci were detected in non-neoplastic oral cells at 4 weeks of 4NQO administration. In the same way, GSTP positive cells were detected in pre-neoplastic lesions and squamous cell carcinomas induced after 12 and 20 weekstreatment, respectively. None of the control animals expressed GST-P positive cells. Regarding cellular proliferation, PCNA positive nuclei were higher at 12 and 20 weeks following 4NQO exposure (p<0.05) when compared to negative control. Conclusion. These results suggest that the expression of GST-P is correlated with cellular proliferation, in which GST-P is associated with risk and progression of oral cancer, whereas PCNA is closely involved during neoplastic conversion.

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PP3-223 DIAGNOSTIC VALUE OF CYTOKERATIN MARKERS IN DIFFERENTIAL DIAGNOSIS OF SALIVARY GLAND TUMORS Mohamad Javad Ashraf1, Neda Bagheri1, Mohamad Shaghasemi1, Negar Azarpira2 1 Shiraz university of Medical Sciences, Iran 2 Transplant Research Center, Iran Inroduction: Salivary gland tumors have one of the most complex histopathologic features of any organ/tissue. In this study we want to evaluate diagnostic value of various cytokeratin markers and smooth muscle actin in differential diagnosis of pleomorphic adenoma (PA) and adenoid cystic carcinoma (ACC). All of the cases had both typical and atypical areas and we suppose that pattern of positivity of cytokeratin markers with SMA in atypical areas can help in proper diagnosis of poorly differentiated tumors. Material and Methods: We selected 35 cases of salivary gland tumors from the pathology files of Khalili Hospital between 2002-2006. These cases were 19 PA, 16 ACC .We performed immunohistochemical staining with six cytokeratin markers (CK7, CK8, CK13, CK14, CK17 and CK18) and SMA. Positivity was defined if 10% or more of the cells were immunostained. Result: In PA group CK7 and CK8 showed 100% positivity in both typical and atypical area. Positivity of CK13 in typical area was 0% but in atypical area was 10.5%. SMA immunoreactivity in typical and atypical areas was 36.8% and 21% respectively (P-value 0.003). 100% of cases showed CK18 positivity in typical area whereas 42% of cases in atypical area were positive.57.9% of cases showed CK17 positivity in typical area and 26.3% in atypical area . In ACC group 87.5% of cases showed SMA positivity in typical areas and 62.5% in atypical areas . CK7 positivity was present in 93.5% of cases in typical areas and 93.8% in atypical areas (p-value = 0.00). 100% of cases revealed CK8 positivity both in typical and atypical areas. None of the cases showed immunoreactivity for CK13 in typical or atypical areas. 93.8% of cases showed immunoreactivity for CK14 in typical areas and 81.3% in atypical areas. 31.3% of cases revealed positivity for CK17 in typical areas whereas none of the cases showed positivity in atypical areas. Conclusion: The following conclusions were obtained: 1- Five markers, SMA, CK7, CK8, CK13, CK14 have significant correlation between typical and atypical areas of both pleomorphic adenoma and adenoid cystic tumors. 2- Positivity of SMA and CK8 is more infavor of ACC rather than pleomorphic adenoma. 3- Coexpression of CK7 and CK14 and simultaneous negativity of CK13 and CK18 are significantly more infavor of pleomorphic adenoma than adenoid cystic carcinoma. PP3-224 METACHRONOUS DEVELOPMENT OF NASAL KAPOSI'S SARCOMA AND MALIGNANT MELANOMA IN A NON AIDS-RELATED PATIENT Ioannis Venizelos, Zoi Tatsiou, Lambros Kambas, Despoina Sioutopoulou Department of Pathology, Hippokration Hospital, Thessaloniki, Greece Primary malignant neoplams of the nasal cavities is believed to comprise about 1% of all human malignancies. Moreover, nonsquamous cell malignant neoplasms are very infrequent, comprising about 0.5% of all malignant tumors in this site. Among the few reported cases of Kaposi’s sarcoma (KS) involving the head and neck region, only 6 had primary presentation in the nasal cavity and 5 of them were related to the acquired immune deficiency syndrome (AIDS). Nasal malignant melanomas (MM) are rare, accounting for less than 1% of all MM. Metachronous development of nasal KS and MM in a nonAIDS related patient has not been reported. We report the case of a 59-year-old woman who was admitted in our hospital due to

rhinorragia. On physical examination it was found a purplish tumor with maximum diameter 0,8 cm mostly obstructing the left nasal cavity and arising from the nasal septum. There was no peripheral lymphadenopathy, hepatosplenomegaly or skin lesions. Hematological examination and chest x-ray was normal. All laboratory tests for bacteria, viruses, HIV, fungi, parasites as well as complete immunological studies for autoimmune diseases were negative. Histological examination of the excised tumor revealed features of KS. Immunohistochemical examination showed positivity of the neoplastic cells for CD34 whereas they were negative for S100, HMB45, Melan A, EMA, SMA and CD45. The patient received no further treatment and 6 years later she was readmitted due to difficulty in breathing. On rhinoscopy a polypoid tumor was found in the left nasal cavity which was considered to be a relapse. The tumor was excised within free surgical margins, it had 3,2 cm maximum diameter and a dark colored cut surface. Histological examination showed features of MM. Immunohistochemicaly, the neoplastic cells were positive for S100, HMB45 and Melan A whereas they were negative for CD34, CD45, SMA and EMA. The patient received no further treatment and 1 year later she is in an excellent condition with no evidence of local recurrence or metastasis, both clinically and radiologically. In the present study we report a unique case of primary KS of the nasal cavity with metachronous development of MM in the same location, in a patient with adequate immune system. PP3-225 A CASE OF CERVICAL ECTOPIC MENINGIOMA: CHALLENGING IN FROZEN SECTION Mohammad Hossein Sanei, Nezamaldin Berjis, Parvim Mahzouni, Azar Naimi Isfahan University of Medical Sciences, Iran Background Primary extracranial and extraspinal meningiomas are rare.they are usually limited to the head and neck region or to the paravertebral soft tissues. histologic diagnosis (particularly of fresh specimens) is often difficult. Case report A 16-year-old girl presented with a cervical mass.It was a sizable mass with an approximate diameter of 5 cm.It was diagnosed in fine needle aspiration as a pleomorphic adenoma. The diagnose of surgon,during the operation was paraganglioma and frozen section suggested an infiltrative tumor. Histologically, the lesion showed uniform spindle cell proliferation separated by hyalinized collagen bundles . The spindle cells were often arranged in sweeping fascicles and concentrically wrapped in tight whorls. Immunohistochemically, tumor cells were positive for EMA and vimentin,while showed negativity for cytokeratin, chromogranin,NSE and thyroglobuline. A diagnose of ectopic meningioma was established based on microscopic appearance and immunohistochemical profile of tumor. Conclusion Although rare,ectopic meningioma should be considered in differential diagnosis of any mass lesion in head and neck region,comosed of spindle cells. PP3-226 THE Ets-1 TRANSCRIPTION FACTOR IS INVOLVED IN PTERYGIAL ANGIOGENESIS Wallid Naib-Majani1, Winrich Breipohl1, Elham Elshazli2 1 Department Of International Medical Education and Development University Of Bonn Germany 2 Research Institute Of Ophthalmology, Egypt Introduction : Pterygial pathology is characterized by abnormal corneal epithelial proliferation, stromal modulation, matrix degradation, and a strong tendency for otherwise absent corneal vascularization. As the proto-oncogene c-ets1 is known to play a key role in angiogenesis and matrix degradation in other tissues. If its involvement in corneal vascularization was to be checked. Method: 25 pterygia in two Groups. Group 1 consisted of five

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clinically active, group 2 consisted of 20 samples clinical non active pterygia. were investigated with 35 S-labeled ets-1 antisense and sense in-situ hybridization of c-ets-1 transcription factor. Results: The cytoplasm of blood vessel endothelial cells showed strong expression of c-ets-1 mRNA in all group 1 pterygia. In contrast, no expression of c-Ets-1 or c-ets-1 was found in the pterygia of group 2. Conclusion: Proto-oncogene cets 1 expression has been shown for the first time in the metaplastic pterygium, an eye tissue of unknown pathogenesis. PP3-227 ERDHEIM-CHESTER DISEASE, A CASE REPORT AND REVIEW OF LITERATURE Maysa Al-Hussaini1, Eman Hijazi2 1 King Hussein Cancer Center, Jordan 2 Private Sector, Jordan Erdheim-Chester disease (ECD) is a rare form of non-Langerhans histiocytosis of unknown origin, characterized by adult onset with multi-systemic involvement, in which diffuse sclerosis of the diametaphysis of the long bones, especially in the lower limbs, is the most consistent finding. Orbital involvement is a rare feature seen in less then a third of cases. we are presenting a case of a 42 year old lady, who presented initially to the ophthalmology clinic with a 2 year history of progressive marked proptosis of the left eye, complicated by complete loss of vision and pain of 2 weeks duration. There was mild proptosis of the right eye, the vision however was preserved. enucleatin of the left eye revealed the presence of histiocytosis of non Langerhans cell type. Discussion of this rare form of presentation of ECD as well as review of the literature is presented. PP3-228 DERMATOFIBROSARCOMA OF HEAD AND NECK. PATHOLOGIST POINT OF VIEW Nadia Kourda, B Tayahi, A Landoulsi, M Helali, F Farah, J Kourda, O Lamine, M Ferchiou, A Adouani, Sarrah Baltagi Ben Jilani, R Zermani Charle Nicolle’s Hospital Department of Pathology, Tunisia Background : Dermatofibrosarcoma protuberans (DFSP) is a rare locally aggressive malignant neoplasm. It is most commonly found on the trunk and rarely on the head and neck. In fact only 12% to 15% of the cases are localized in the head and neck region. It represents 1.4% of the soft tissue sarcomas of this area, and its main feature is the high rate of recurrence. This study defines the clinical behavior of DFSP evaluates the role of pathologist in surgical margins analysis, and identifies factors that predict local control. Methods: Hospital records and pathological slides were reviewed for 5 patients with pathologically confirmed head and neck (DFSP) treated at Charles Nicolle Hospital between 1999-2006. Results: This study included 5 cases (3 men and 2 women; mean age 38 years, (range 28–48 years) of DFSP. 4 cases were primary tumors and 1 was recurrences after surgery excision. The tumors were located in the following areas: the scalp in two cases, the cheek in three cases. In all cases clinical and instrumental examinations showed absence of regional and distant metastases. Tumor size ranged from 4 cm to 10 cm. All 5 subjects were treated by surgical excision of the tumor. 2 our 5 subjects cases were with positive margin. These 2 patients had a recurrence of the disease, after 12 months of follow up. The new lesion was treated by resection with a 5-cm margin. Conclusions: Wide margin resection of head and neck DFSP predicts negative histological margins and impacts favorably on local recurrence-free survival.

PP3-229 OSTEOPLASTIC TRACHEOPATHY Simion George1, Ilie Marius1, Chirculescu Florin2, Marinescu Elena1, Sajin Maria1 1 Department of Pathology, Emergency University Hospital Bucharest, Romania 2 Department of Cardiothoracic Surgery, Emergency University Hospital Bucharest, Romania Background: Osteoplastic tracheopathy is a very rare disease that presents itself as multiple submucosal nodules composed of mature bone and cartilage. Method: We report the case of a 35year-old male diagnosed with schizophrenia 12 years ago and tried to attempt suicide by sedative ingestion. These induced comatose (GCS = 5), which required oro-tracheal intubation. The patient recovered from comatose and stabilized within 4 days. 4 months later the patient presents respiratory distress that accentuates progressively. He’s diagnosed with hypercapnic acute respiratory failure (PCO2 = 120 mmHg) and subglotic tracheal stenosis. The fiberoptic bronhoscopic procedure reveals a 2.5 cm in depth / 1 cm in width subglotic tracheal stenosis, caused by multiple partially necrotic granulomas, developed mainly on the left lateral wall. After a moderate hemoptoic episode and the aggravation of respiratory troubles surgery is considered necessary: tracheal subcricoid segmentary resection with termino-terminal anastomosis. The method used for diagnose was paraffin-embedded tissue and stained with hematoxylin and eosin (H&E). Results: Histopathological out findings revealed the presence of bone and cartilage nodules and plaques mainly in subepithelial tissues, underline tracheal epithelium acanthosis and papilomatosis with richly vascularised stromal repairing tissue. Only isolated epithelial ulcerations are seen. Conclusions: In this case the lesion’s etiology seems to be inflammatory, as a consequence of the oro-tracheal intubation. PP3-230 HISTOPATHOLOGY OF FLOOR OF MOUTH LEUKOPLAKIA: A MIMIC OF ORAL LICHEN PLANUS? Séamus Napier, Jacqueline James, Rajeev Shah Histopathology, Royal Hospitals, Belfast, Northern Ireland Background: Oral lichen planus (OLP) is reputed to have characteristic clinical and histopathological appearances that should allow confident diagnosis. What is not well recognised is that the histopathological features of OLP may overlap with changes due to tobacco use. The floor of mouth (FOM) is rarely affected by OLP but can develop leukoplakia in smokers. Defining the spectrum of histopathological findings of FOM leukoplakia biopsies will highlight the proportion of tobaccoassociated lesions that exhibit changes similar to those seen in OLP. Method: Laboratory records were trawled for FOM biopsies from August 2000 to December 2003. When cases of carcinoma, dysplasia and other “non-leukoplakia” diagnoses were excluded, 59 suitable cases were identified. Original slides of these 59 patients were reviewed by an oral pathologist blinded to the initial diagnosis and the following features recorded semiquantitatively: degree and type of keratosis, altered epithelial thickness, intraepithelial lymphocytes, apoptotic cells, lymphoid follicles and melanin pigmentation. The findings were compared to the clinical summary and smoking history. Results: A tobacco history was recorded for 53 (90%) of patients: 50 were current smokers, 3 were “non-smokers”. No smoking history was available for 6 (10%) patients. All cases exhibited hyperkeratosis and alterations of epithelial thickness, most often a mixture of ortho- and parakeratin with adjacent areas of atrophy and hyperplasia. Intraepithelial lymphoctyosis was seen in 49 (83%) cases, of which 44 (75%) were known to be smokers. Apoptotic cells were present in 35 (59%) cases and were numerous in 9 (15%) cases, of which 33 (56%) were known to be smokers. Abundant melanin pigment accumulation was seen in only 20

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(34%) cases, of which 19 (32%) known to be smokers. Conclusions: The presence of clusters of intraepithelial lymphocytes, apoptotic cells and/or melanin pigment in FOM mucosa is strongly associated with a history of smoking, particularly when abundant. These “interface mucositis”-like changes mimic oral lichen planus and question the validity of such a diagnosis in patients who smoke. PP3-231 IMMUNOEXPRESSION OF PDGF AND PDGFR IN DIFFERENT DEGREES OF CARCINOMA EXPLEOMORPHIC ADENOMA INVASION Cristiane Furuse1, Patricia Ramos Cury1, Albina Altemani2, Venâncio Avancini Ferreira Alves3, Ney Soares De Araújo4, Vera Cavalcanti De Araújo1 1 São Leopoldo Mandic Dental Research Center, Campinas, SP, Brazil 2 State University of Campinas School of Medicine, Campinas, SP, Brazil 3 University of São Paulo School of Medicine, São Paulo, SP, Brazil 4 University of São Paulo School of Dentistry, São Paulo, SP, Brazil Background: Immunoexpression of PDGF and PDGFR was investigated along the progression of pleomorphic adenoma to carcinoma ex pleomorphic adenoma (CXPA) using sixteen cases of CXPA in different degrees of invasion –intracapsular, minimally and frankly invasive carcinoma. Method: To this aim, we counted positive cells in different areas including reminiscent pleomorphic adenoma, in situ and peripheral areas of both intracapsular and minimally invasive tumours as well as in the frankly invasive carcinoma with only epithelial component and/or myoepithelial component. A minimum of 500 cells at 400X in the most marked areas (hotspots) was counted for each site by the computer-assisted image analysis. Results were presented as % of positive stained cells. Results: PDGF and PDGFR were observed in the cytoplasm of the cells but PDGFR was also present in some nuclei. Reminiscent pleomorphic adenoma showed focal stained of PDGF and PDGFR in luminal and plasmacytoid cells and in the cells of myxoid and condroid areas. In situ areas of intracapsular and minimally invasive types showed only strong expression in malignant luminal cells (100%). Both PDGF (71%) and PDGFR (80%) were present in the peripheral areas of intracapsular and minimally invasive with irregular distribution. In frankly invasive ones of the epithelial type, the percentage of positive cells was 90% for PDGF and 80% for PDFGR. In areas with small nests, negative cells were more evident. In frankly invasive type with myoepithelial component, 92% for PDGF and approximately 100% for PDGFR were observed. Conclusion: It seems that the expression of PDGF and PDGFR is involved at the beginning of the malignant transformation since its increase is present in in situ areas when compared with the reminiscent PA. But, their absence in the focal areas in the periphery of the tumours and in some cells of the small groups leads us to the hypothesis that they are not involved in the invasive process what merits further investigation.

PP3-233 ERCC1 IMMUNOHISTOCHEMICAL EXPRESSION IS RELATED TO HISTOLOGICAL DIFFERENTIATION AND PREDICTS OBJECTIVE RESPONSE AND DISEASEFREE SURVIVAL IN PATIENTS TREATED BY CISPLATIN-BASED CHEMOTHERAPY FOR LOCALLY ADVANCED HEAD AND NECK SQUAMOUS-CELL CARCINOMA Adriana Handra-Luca1, Juana Hernandez2, Giannis Mountzios2, Estelle Taranchon2, Jean Lacau-St-Guily3, Jean-Charles Soria4, Pierre Fouret5

1

APHP Hopital Jean Verdier Universite Paris 13/Nord, France Institut Gustave Roussy, France 3 APHP Hopital Tenon, France 4 Institut Gustave Roussy Universite Paris 11, France 5 Institut Gustave Roussy Universite Paris 6, France 2

Background: The excision repair cross-complementation group 1 (ERCC1) enzyme plays a rate-limiting role in the nucleotide excision repair pathway which recognizes and removes cisplatininduced DNA adducts. The relation between ERCC1mRNA expression and resistance to platinum compounds has been studied in advanced-stage gastric, ovarian, colorectal, esophageal, and non-small-cell lung cancer. ERCC1 immunohistochemistry was predictive of the survival benefit of cisplatin-based chemotherapy in non-small cell lung cancer. Purpose: The goal of this study was to examine whether ERCC1 immunohistochemical expression in locally advanced head and neck squamous-cell carcinoma (HNSCC) was correlated to clinico-pathological characteristics and p53 mutations of patients treated with a cisplatin-based chemotherapy. Design: Pre-therapeutic biopsy samples from 96 patients with a known tumor response were evaluated for ERCC1 immunohistochemistry. The median of the H-score (the proportion of stained tumour nuclei multiplied by the staining intensity of nuclei) was used as cutoff. P53 mutations had been previously studied. Results: Patients’ age varied between 51 and 67 years. Tumors were classified as T1/T2 (18) or T3/T4 (78), and nodal involvement was classified as N0 (46), N1 (13), N2 (22) or N3 (15). Of 96 patients, 68 (71%) had tumors that expressed ERCC1 intensively and diffusely, at high levels. Tumors with low ERCC1 were less differentiated than those with high levels (p=0.03). There was no difference for any other characteristics including age, gender, T, N, M, tumor localization or p53 mutation. Among 28 patients with tumors with low ERCC1, 22 (79%) had an objective response as compared to 38 of 68 (56%) patients with tumors with high ERCC1 (p=0.04). Using the logistic regression method, the 28 patients (29 %) with low ERCC1 had a four time greater odds of benefiting from an objective response to chemotherapy (OR 4.3, 95% CI: 1.4–13.4; p=0.01) as compared to the group of 68 patients with high ERCC1. ERCC1 and p53, but not their interaction, were independent predictors of tumor response. In a Cox model adjusted on age, T, N, M, tumor differentiation, and tumor localization, low ERCC1 was associated with a lower risk of cancer death (RR 0.42, 95% CI: 0.20-0.90; p=0.04), while p53 status had no prognostic value. Conclusion: The results of our study suggest that: 1) tumours with low ERCC1 were less differentiated and that, 2) those patients having HNSCC with low ERCC1 are more likely to benefit from cisplatin chemotherapy as compared to patients with high ERCC1. PP3-234 PATHOLOGIC ESTIMATION OF ORAL MUCOSA LESIONS Aphrodite Nonni, Pagona Ctena-Agapitou, Andreas Lazaris, Ilias Papadopoulos, Nikolaos Kavantzas, Emmanuel Agapitos, Sophia Tseleni-Balafouta, Efstratios Patsouris 1st Department of Pathology, Medical School, National and Kapodistrian University of Athens, Greece Background In periodontal destruction, bacterial substances mainly interact with inflammatory cells and stimulate epithelial cells either towards proliferation or death. Actinomycosis is an infectious disease that frequently has chronic granulomatous and suppurative lesions; however, intraorally and periodontally types of actinomyces infection occur rarely. Heat shock proteins (HSPs) are produced by cells as a protection against stress and may play a role in course of infections. CD43 is a cell surface glycoprotein with an intracellular domain implicated in signal transduction and consequent activation of phagocytes. Material and methods Thirty one oral mucosa specimens infected by

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actinomyces were studied with regard to the immunohistochemical expression of HSP70, histiocytic marker CD68 and CD43. Gram stain was performed in all cases. Results The inflammatory reaction consisted mainly of lymphocytes and plasma cells with a focally increased presence of neutrophils. The percentage of macrophages, as assessed by CD68 immunoreactivity, did not exceed 10% of the inflammatory infiltrates. HSP70 was overexpressed in inflammatory cells as well as on actinomyces’ surface and epithelial cells of the basal layer with a concomitant heavy inflammatory reaction. When actinomyces expressed HSP70 strongly, the adjacent epithelium above the basal layer was denuded; on the contrary, when HSP70 expression on actinomyces was low, the neighboring epithelium was hyperplastic. CD43 was expressed mainly in mononuclear inflammatory cells and it was practically negative in neutophils; the latter is probably linked with the increased presence of actinomyces in the examined samples. Conclusions HSP70, when expressed on actinomyces’ surface is likely to interact with adjacent epithelium. The absence of CD43 on neutophils seems to favor the colonization of oral mucosa by actinomyces and the progress of the infection-induced lesions. PP3-235 p53 EXPRESSION IN SINONASAL MUCOSA OF LEATHER WORKERS Annarita Palomba1, Tonina Iaia2, Mauro Biancalani3, Stefano Conti4, Giuseppe Battista5, Milena Paglierani1, Alessandro Franchi1 1 Department of Human Pathology and Oncology, University of Florence, Italy 2 Department of Prevention, USL11, Empoli, Italy 3 Division of Anatomic Pathology, Department of Laboratory Medicine, “S. Giuseppe” Hospital, Empoli, Italy 4 Division of Otolaryngology Head and Neck Surgery, “S. Verdiana” Hospital, Castelfiorentino, Italy 5 Department of Prevention, University of Siena, Italy Background. The association between sinonasal intestinal-type adenocarcinoma and the exposure to leather dusts has been widely documented, but the identification of precursor lesions and early alterations of p53 expression in the exposed nasal mucosa has so far received little attention. Methods. Biopsies of the mucosa of the middle turbinate were obtained from 40 subjects who had been employed in leather factories for 9-48 years (median 27 years). Thirty-three subjects had been employed in leather tanning activities, 21 in shoe making activities, and 14 had been exposed to both. Tissue samples were routinely processed for histologic examination and immunostaining for p53 was performed using the monoclonal antibody DO7 (Dako Co., Glostrup, Denmark). For statistical analysis, cases showing >20% of positive cells were considered positive. Results. In 22 biopsies (55%) we observed nuclear positivity for p53 in surface nasal epithelium. Immunoreactivity was present in basal and suprabasal epithelial cells in squamous metaplastic epithelium, while native schneiderian epithelium and goblet cells were negative in most cases. Concerning glandular epithelia, positivity was detected in 35 samples, 8 of which showed >20% of positive cells. Overall, p53 expression in surface epithelia was more frequently found in subjects employed in leather tanning than in subjects employed in shoe making or in both activities (p=0.06, Fisher’s exact test). p53 expression in surface epithelia was significantly associated with longer exposure to leather tanning activities (p=0.02, Mann Whitney U test), while no correlation was found with shoe making activities, or with global work exposure. Similarly, no correlation was found between p53 expression in glandular epithelia and work exposure parameters. Conclusions. In nasal mucosa, a site at high risk of developing intestinal type adenocarcinoma in subjects exposed to leather dust, epithelial cells frequently overexpress p53 protein, and this appears to be related particularly to leather tanning activities.

Further molecular studies are needed to clarify the role of p53 overexpression in the pathway leading to the development of these tumours in exposed subjects. PP3-236 THE ENAMEL MATRIX DERIVATIVE (EMDOGAIN®) ENHANCES HUMAN TONGUE CARCINOMA CELLS GELATINASE PRODUCTION, MIGRATION AND METASTASES FORMATION Matti Laaksonen1, Juho Suojanen1, Sini Nurmenniemi2, Esa Läärä3, Timo Sorsa1, Tuula Salo2 1 Department of Oral and Maxillofacial Diseases, Helsinki University Central Hospital, Institute of Dentistry, University of Helsinki Helsinki, Finland 2 Department of Diagnostics and Oral Medicine, Oulu University Central Hospital, Institute of Dentistry, University of, Oulu, Oulo Finland 3 Division of Statistics, Department of Mathematical Sciences, Faculty of Science, University of Oulu, Oulu, Finland Background: Enamel matrix derivative Emdogain® (EMD) is widely utilized in periodontal therapy to regenerate lost connective tissue and to improve the attachment of the teeth. Gelatinases, matrix metalloproteinases -2 and -9, are supposed to have an essential role in the progression of oral cancer growth and metastasis formation. The aim of this study was to determine the effects of EMD on human tongue squamous carcinoma (HSC3) cells in vitro and in vivo. Methods: Subconfluent cultures of HSC-3 cells and spontaneously immortalized oral mucosal keratinocytes (HMK) were incubated for 48 h in serum-free culture media containing 0, 100 or 200 μg/ml of EMD. Gelatinase production was analyzed using gelatine zymography and enzyme-linked immunosorbent assay (ELISA) kits. For transwell cell migration assay both cell lines were pre-incubated with EMD and allowed to migrate overnight. The attached cells on the lower side were stained and quantitated. In a set of experiments, a cyclic anti-gelatinolytic CTT-2 peptide was added to the transmigration assay. Cell proliferation was analyzed using microtiterplates coated with or without EMD. The cells were incubated from 12 to 96 h. The proliferation was determined using Cell Proliferation ELISA BrdU kit. For in vitro wound closure assay BSA pre-coated confluent cell culture plates were wounded and exposed to various concentrations of EMD. After incubation the wound areas were measured. For in vivo metastasis assay HSC-3 cells were injected subcutaneously into the back of athymic nude mice. After three days of inoculation, mice were injected subcutaneosly with or without EMD daily during five days of treatment. After scarification the number of the metastases were counted. Results: EMD at concentrations of 100 μg/ml and 200 μg/ml significantly induced the production of MMP-2 and -9 from HSC-3 cells. EMD also slightly induced the gelatinase production from HMK cells. Furthermore, EMD induced the transmigration of HSC-3 cells, but reduced the migration of HMKs in transwell assays. The in vitro wound closure of HSC-3 cells was significantly accelerated by EMD, whereas it did not effect on the wound closure of HMKs. The migration of both cell lines was inhibited by a specific cyclic anti-gelatinolytic peptide CTT-2. EMD had no effect on HSC-3 cell proliferation. EMD significantly induced metastases formation, but had only a limited effect on mice survival bearing human tongue HSC-3 carcinoma xenografts. Conclusion: We suggest that the use of EMD for patients with oral mucosal malignancies should probably be avoided.

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PP3-237 A COMPARISON BETWEEN THE FREQUENCY RATE OF MALIGNANT & PREMALIGNANT WHITE ORAL LESIONS AND THE REST OF WHITE ORAL LESIONS REFERRED TO THE RECORDS OF TEHRAN UNIVERSITY (IRAN) PATHOLOGY CENTER Parisa Piroozmand, Zahra Hosseini, Donya Sadri School of Dentistry, Islamic Azad University, Tehran, Iran BACKGROUND: White lesions of oral mucosa is caused by thickening of keratin layer, epithelial hyperplasia, intraepithelial edema and reduced vascularity in the underlying lamina propria that in some cases transformation into malignancy may occur. The aim of this study is to assess knowledge of potentially malignant white oral lesions indices rates, in order to prevent oral cancer. METHODE: In this descriptive study a total of 4799 documents of patients referred to the records obtained from the pathology section of Tehran University were reviewed from the year 1995 to 2005. Documents of affected patients by white oral lesions were evaluated. Variables (sex, age, lesion type, location of oral involvement, microscopic diagnosis) were assessed by Chi-Square analytical test. RESULTS: Of our samples, 425 patients (9%) were affected by white oral lesions, of them, 7.06% were premalignant and 22.35% were malignant and the rest (70.59%) had no report of malignancy and were benign. Among premalignant white lesions idiopathic leukoplakia with mild dysplasia had maximum percentage. Based on the results of this study, the maximum percentage of premalignant lesion occurrence were reported to be among the males over the age of 50 on the buccal mucosa vs. malignant lesions which were predominantly reported in females over the age of 50 on the tongue. CONCLUSION: Obtained results showed that 9% of oral lesions are white with less prevalence rate compared with that obtained in previous studies. This may arise from deficit consideration and carelessness of dental staff during examination and exclusion of white lesions. However it is found that 22.35% of white lesions are malignant, bigger percentage than that obtained by similar investigations. This may also arise from less consideration in primary diagnostic approaches performed by dentists. Considering prevalence of premalignant and malignant lesions among elderly patients, paying more attention during clinical examination is mandatory particularly for high risk patients. PP3-238 INTRAMUSCULAR HEMANGIOMA OF THE RETROPHARYNGEAL SPACE. A CASE REPORT Ivan Boricic1, Zorica Stojsic1, Dimitrije Brasanac1, Nada Tomanovic1, Dragoljub Bacetic1, Anton Mikic2 1 Institute of Pathology, School of Medicine University of Belgrade, Serbia 2 Institute of Otorhinolaryngology, School of Medicine University of Belgrade, Serbia Intramuscular hemangioma is a distinctive type of benign vascular tumor confined to the skeletal muscle. This tumor is prone to recur and behaves in a locally aggressive manner. The most common site of involvement is the lower limb, followed by the head and neck region. To our knowledge, only one case of intramuscular hemangioma arising in the retropharyngeal space has been reported in the English literature. We, herein, report a further case in a 51-year-old woman. The tumor manifested as a left-sided neck swelling of 15-day duration. The patient was found to have bulging of the posterior oropharyngeal wall on indirect laryngoscopy and an ill-defined left-sided retropharyngeal mass on computed tomography scan. The tumor was excised by a transoral approach and microscopically diagnosed as intramuscular hemangioma, the complex malformation subtype. It was mainly composed of irregularly shaped large- and medium-sized blood vessels, reminiscent of

arteries and veins. The second vascular component corresponded to capillaries, lined by prominent endothelial cells. Vascular channels were set in a fibrous-fatty stroma with entrapped striated muscle fibers. In addition, isolated bundles of smooth muscle fibers were present throught the tumor. Some of these structures contained lumens and were attached to vessel walls. We assume that at least some of these smooth muscle bundles could be malformed blood vessels, as well. Intramuscular hemangioma should be considered in the differential diagnosis of any soft tissue mass found in the retropharyngeal space. PP3-239 THE EXPRESSION OF Ets, E-CADHERIN AND Ki-67 IN HEAD AND NECK SQUAMOUS CELL CARCINOMA. AN IMMUNOHISTOCHEMICAL STUDY Paul Nagy1, Barnabás Horváth2, Ferenc Salamon1, Zsuzsa Schaff3 1 National Medical Center, Department of Pathology , Hungary 2 National Medical Center, Department of Otorhinolaryngology , Hungary 3 Semmelweis University, 2nd Department of Pathology, Hungary Background: Despite the development of diagnostic tools and therapy, squamous cell carcinoma of the head and neck (HNSCC) may present challenges to pathologists and clinicians as well. A particularly difficult problem is the treatment of regional metastases. We examined immunohistochemically molecules involved in the metastatic process: ets-1 transcription factor that regulates expression of matrix metalloproteinase-3, E-cadherin that is responsible for homotypic cell-cell adhesion, and cell proliferation marker Ki-67. We studied their possible relations relevant to the clinical practice. Method: 98 patients (87 males and 11 females) with HNSCC were treated only surgically in our institute during a period of 3 years . We divided the primary tumor samples into two groups. Group N(0) (n=49) were selected from patients without synchronous lymph node metastasis or nodal recurrence in a follow-up period of 2 years and Group N(+)(n=49) were from patients with lymph node metastases. Using immunohistochemistry the targets mentioned above were detected in paraffin-embedded specimens representing the primaries and the metastases as well. The following clinicopathological parameters were taken into account: sex, age, tumor localization, grade and stage. The results were statistically analyzed. Statistical significance was defined as p<0.05. Results: Correlation between ets-1 expression and clinicopathological setting, metastasis and Ki-67 labeling index (LI) was not significant. Reduced expression of E-cadherin showed significant correlation with metastasis, survival and Ki-67 LI. In addition, we found that the degree of E-cadherin expression in primary tumors may differ from that of the metastases. Conclusion: Ets-1 can be demonstrated mainly in the peritumoral stromal cells and has no direct prognostic significance. However, testing the Ki-67 labeling index and E-cadherin expression may be useful in identifying high-risk patients who need more aggressive therapy. Moreover, these markers may be valuable in a panel of prognostic factors for HNSCC. PP3-240 ANGIOGENIC SWITCH DURING TUMOR PROGRESSION OF CARCINOMA EX-PLEOMORPHIC ADENOMA (CXPA) Andresa Soares1, Priscila Juliano1, Vera Araujo2, Randal Adam1, Konradin Metze1, Albina Altemani1 1 State University of Campinas - Brazil 2 São Leopoldo Mandic - Brazil BACKGROUND: Angiogenesis is considered to be a critical process for tumor growth, invasion and metastasis. CXPA is the principal form of malignancy arising in pleomorphic adenomas (PA) and is usually a high-grade neoplasm, with frequent

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metastases and a poor clinical outcome. We investigated the question whether an angiogenic switch would take place during the malignant transformation of PA into carcinoma and during tumor progression. METHOD: In 8 cases of early CXPA (intracapsular and minimally invasive), 8 of advanced CXPA (widely invasive tumors) and 10 cases of PA without malignant transformation, tumor vascularisation was assessed by measuring total microvascular area (TVA) and microvessel density (MVD) in CD34 and CD105 stained sections. In digitalized images the texture of the CD34 stained capillaries was analyzed calculating Shannon´s entropy (as a measure of the CD34 staining heterogeneity) and the fractal dimension, which represents the complexity of the architecture. RESULTS: Whereas the CD34 values did not differ among the tumors, MVD for CD105 increased significantly during tumor progression (PA without malignant transformation: mean 3.3; early CXPA: 9.0; widely invasive CXPA: 33.6) with carcinomas without myoepithelial differentiation revealing higher values (mean 46.5) than carcinomas with such cellular differentiation (mean 16.3). In relation to TVA, in advanced CXPAs the TVA value was higher (mean 179.4) than in early carcinomas (mean 63.8) and tumors with myoepithelial differentiation showed higher values (mean 260.2) than those without (mean 118.9). In widely invasive tumors with myoepithelial differentiation the carcinoma cells often formed large hypovascularized aggregates surrounded by large vessels whereas in those without myoepithelial differentiation the cell aggregates were smaller with thinner vessels around them. Entropy and the fractal dimension raised during carcinogenesis (lower in adenomas and higher in frankly invasive carcinomas), which is equivalent to an increase in heterogeneity of the endothelial CD34 expression and complexity of the vessel architecture. CONCLUSION: During the progression from adenoma to carcinoma there is an increasing complexity of the vessel architecture. The antibody CD105 reflects better the angiogenic switch than CD34. The degree of angiogenesis and the total vascular area have distinctive patterns in CXPA with and without myoepithelial differentiation. Low angiogenesis associated with high TVA value is more characteristic of CXPA with myoepithelial differentiation. FAPESP-04/07960-0 PP3-241 POTENTIAL PROGNOSTIC IMPORTANCE OF THE EXPRESSION OF MATRIX METALLOPROTEINASE MMP9 AND TISSUE INHIBITOR OF METALLOPROTEINASE TIMP2 IN PATIENTS WITH LARYNGEAL SQUAMOUS CELL CARCINOMA Jozef Kobos1, Elzbieta Salacinska-Los1, Wioleta Pietruszewska2 1 Department of Pathology of the Age of Development Medical University of Lodz, Poland 2 Department of Otolaryngology Medical University of Lodz, Poland Matrix metalloproteinases belong to the family of human endopeptidases involved with degrading of the extracellular matrix. MMP9 belongs to the group of matrix proteins called gelatinases and TIMP2 acts as an inhibitor of metalloproteinases and also may promote growth inhibition. The aim of the study was the evaluation of the immunoexpression of matrix metalloproteinase MMP9 and tissue inhibitor of metalloproteinase TIMP2 in patients with laryngeal squamous cell carcinoma. Our material consisted of 67 cases of laryngeal squamous cell carcinoma (LSCC) from the files of the Department of Pathology Medical University of Lodz. For immunohistochemical study we used Novocastra Company antibodies and EnVisio (DAKO) visualisation system. The MultiScanBase CSS Ltd. Company image analysis system was used for scoring the intensity of expression of evaluated proteins. Only tumor cells with distinct cytoplasmic immunoexpression were classified as positive. Our date showed that the significantly

higher number of TIMP2 positive cells were observed in grade three LSCCs than in low and intermediate grade tumors. We also noted that the intense MMP9 expression was present more frequently in patients with high grade carcinomas but the results were not significant. We seen the MMP9 positive neoplastic cells mostly at the margins of tumor areas adjacent to the stroma. We also found the positive correlation between MMP9 and TIMP2 expression in evaluated carcinomas. In conclusion we believe that the TIMP2 and MMP9 expression might be useful prognostic markers in patients with oral squamous cell carcinoma. PP3-242 PLEOMORPHIC ADENOMA (MIXED TUMOR) OF THE MINOR SALIVARY GLANDS Christos Makos1, Anastasia Nikolaǿdou2, Rodi Kotakidou3, Symela Amanatidou4 1 Department Of Oral And Maxillofacial Surgery, General Hospital Of Kilkis, Greece 2 Department Of Pathology, General Hospital Of Kilkis, Greece 3 Department Of Pathology, General Hospital “G. Gennimatas’’, Thessa-Loniki, Greece 4 Technisian, Greece BACKROUND: Pleomorphic adenoma is an epithelial tumor of the salivary glands. The first prescription was made by Billroth and Von Bruns in 1859 and the initial name “mixed tumor” was given by Minssen in 1874 as a manifestation of a mixed epithelial and mesenchymal origin. Today, the prevalent name is pleomorphic ade-noma because of the proved isolated epithelial origin and the pleomorphic histological picture per location. The frequency is estimated about 60-80% of the salivary gland tumors and a 90-96% of the benign salivary gland tumors with a 85-90% of the cases located in the parotid glands and mainly to the distal caudal section. A pleomorphic adenoma in the minor salivary glands is not often. The purpose of this work is to pre-sent cases of pleomorphic adenomas of the minor salivary glands clinically presented as sebaceous cysts and or lipomas and to discuss about this entity. METHODS-RESULTS: The clinical, surgical and histological pictures in cases with pleomorphic adenomas of the minor salivary glands are presented and the particulari-ties in histological sections and stains are referred. CONCLUSION: Definite diagnosis in the above mentioned pleomorphic adenomas of the minor salivary glands is emphasized by histological evidences. Because of the relative small size of the tumor and the expansion in the cheek and labial subcutane-ous tissue these tumors clinically could be misdiagnosed as sebaceous cysts, pilar cysts and or lipomas. PP3-243 HUMAN PAPILLOMAVIRUS PRESENCE AND ASSOCIATED RISK WITH LARYNGEAL NEOPLASIAS Anastasia Nikolaidou1, T. Balampanidis2, I. Venizelos3, D. Rizos2, T. Apostolidis2, K. Tsimaratou4, I. Pappa4, A. Kotsinas4, V. Gorgoulis4 1 Department of Pathology, General Hospital of Kilkis, Greece 2 Department of Otorinolaryngology Head and Neck Surgery, General Hospital of Kilkis, Greece 3 Department of Pathology, General Hospital ‘’Ippokratio’’, Thessaloniki,Greece 4 Molecular Carcinogenesis Group, Department of Histology and Embryology, School of Medicine, University of Athens, Greece BACKROUND: The purpose of this study was to determine the relationship between human papillomavirus (HPV) infection in patients with invasive squamous cell carcinoma, in situ carcinoma, keratosis and mild-severe dysplasia of the larynx. HPV has been implicated as a risk factor in this carcinogenic process after cigarette smoking and chronic alcohol consumption. MATERIALS & METHODS: Formalin-fixed, paraffinembedded tissue was obtained from 20 males (aged 60-79) with

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laryngeal neoplasias. Extraction of total DNA from sections was performed using the QIAamp DNA mini kit (QIAGEN). Quality and integrity of extracted DNA was tested by PCR amplification of an amplicon of the IFN-Ȗ gene. A nested, multiplex, highly sensitive PCR method (~1fg; 103 viral copies) was used for HPV detection and genotyping. In this assay consensus primers for first-run amplification of a broad spectrum of mucosal HPV genotypes, including all high-risk HPV genotypes, were combined with nested PCR amplifications of type-specific primers. RESULTS: Out of the 20 samples, two of them (10%) were found positive for the high-risk subtype HPV16 (case 3, invasive carcinoma, moderately differentiated) and the low risk subtype HPV43 (case 16, invasive carcinoma, well-moderately differentiated), respectively. CONCLUSION: Both high and low risk HPVs (HPV16 and HPV43, respectively) were detected in patients with invasive laryngeal squamous cell carcinomas. The presence of, especially, high-risk HPV genotypes may identify a subset of patients at risk to develop invasive laryngeal squamous cell carcinoma. Nevertheles, examination of larger numbers of patients are required to expand these findings. PP3-244 IMMUNOHISTOCHEMICAL DETECTION OF ECADHERIN AND BETA-CATENIN IN SALIVARY GLAND TUMORS Theodoros Papadas, Nikolaos Mastronikolis, Maria Gkermpesi, Panagiota Ravazoula University Hospital of Patras, Greece BACKGROUND & OBJECTIVE: E-cadherin links to the cytoskeleton via catenins and mediates cell-cell homophilic adhesion. Beta-catenin not only regulates cell-cell adhesion, as a protein interacting with cadherin, but also functions as an important component of the Wnt signaling pathway which has been found to be closely associated with tumor formation. This study was performed to examine the expression of E-cadherin and beta-catenin in salivary gland tumors in order to evaluate their possible roles in the formation of salivary gland tumors. MATERIAL & METHODS: Archival formalin-fixed, paraffinembedded tissue sections of 48 mixed tumors (pleomorphic adenomas), 17 Warthin tumors, 5 mucoepidermoid carcinomas, total 70 tumors, were studied immunohistochemically using an Envision/horseraddish peroxidase (HRP) technique. Healthy salivary glands were used as controls. RESULTS: Membrane and cytoplasmic associated E-cadherin and beta-catenin expression was present in 54/70 and 68/70 of the tumor types studied, respectively. E-cadherin and beta-catenin showed a similar distribution; however, E-cadherin was less frequently expressed than beta-catenin. Expression of both proteins was stronger in Warthin tumors and in mucoepidermoid carcinomas. Reduction and/or absence of E-cadherin were only observed in mixed tumors. CONCLUSION: This study suggests a direct association of E-cadherin and beta-catenin expression with neoplastic histology phenotype of salivary gland tumors. PP3-245 EXPRESSION OF METALLOPROTEINASES IN SALIVARY GLAND TUMORS Maria Gkermpesi, Nikolaos Mastronikolis, Theodoros Papadas, Dimitra Koumoundourou, Panagiota Ravazoula University Hospital of Patras, Greece Matrix metalloproteinases (MMPs) are involved in extracellular matrix turnover. MMPs belong to a family of zinc-dependent endopeptidases and are classified according to their substrate specificity. Although MMPs are among the potential key mediators of cancer invasion, their involvement in premalignant lesions is not clarified. The aim of our study was to evaluate the expression of gelatinase A (MMP-2) and gelatinase B (MMP-9) in salivary gland tumors. Material and Methods: We examined

immunohistochemicaly formalin-fixed, paraffin-embedded tumor samples from 70 patients (48 pleomorphic adenomas,17 warthin tumors and 5 mucoepidermoid carcinomas) for MMP-2 and MMP-9 (Chemicon Laboratories) expression. Results: Cytoplasmic MMP-2 staining was detected in epithelial component of all pleomorphic adenomas and warthin tumors, and also in stoma fibroblasts and endothelial cells. MMP-9 cytoplasmic staining was detected in myoepithelial cells of pleomorphic adenomas, in epithelial cells of warthin tumors and in inflammatory cells in stroma. Nuclear immunostaining for MMP-9 was also detected in epithelial component of pleomorphic adenomas. In mucoepidermoid carcinomas MMP-2 and MMP-9 were also immunoexpressed in squamous malignant component. Conclusions: Our results suggest that MMP-2 and MMP-9 expression is an early event in salivary gland tumorogenesis. Our findings suggest roles of MMPs in tumor initiation in addition to invasion. PP3-246 DESMOPLASTIC FIBROBLASTOMA OF THE LARYNX Fred Ellinger1, Karla S. Pereira1, Adriana Brentegani1, Fernando Siemann1, Thiago Y. Miura1, Silvio B. Uvo1, Luiz C. Silva1, Frederico G. N. Ellinger1, Christopher D. M. Fletcher2 1 FAMEMA, Brazil 2 Brigham and Women’s Hospital, United States BACKGROUND Desmoplastic fibroblastoma (DM) is a rare benign soft tissue tumor of fibroblastic origin, first described by Evans in 1995. It affects mainly adult males, most commonly limbs and trunk and has no recurrences. It is characterized by a paucicellular proliferation of stellate shaped fibroblasts in a collagenous or myxoid background. Two cases have been described in the palate and one in the parotid gland. The authors describe the first case in the larynx. METHOD Representative slides were stained by hematoxilin and eosin and immunohistochemistry study was conducted using the Envision Plus detection system (Dako, Carpinteria CA) according to well established technique. Clinical and follow-up information were obtained from the files of the clinician. RESULTS Case presentation: ASA, 51 years old male presented complaining of “sore throat” and a foreign body sensation. Direct laryngoscopy reavealed a two centimeters, deglutation mobile nodule, in the larynx. It was locally ressected. Grossly the nodule was whitegrayish, firm, well demarcated, measuring 1,9 X 1,9 X 1,6 cm. Microscopically there was a well demarcated hypocellular nodule, underneath a squamous mucosa, composed of proliferating spindle and stellate fibroblasts. The stroma was collagenous and focally myxoid. The diagnosis was desmoplasic fibroblastoma. Because of the unusual location of the lesion, the case was sent in consulation to Dr. C.D.M. Fletcher who agreed with the diagnosis. He also performed and immunohistochemical study. The lesional cells were focally positive for CD-34, while negative for smooth muscle antigen (SMA) and S-100 protein. DISCUSSION AND CONCLUSION Desmoplastic fibroblastoma is a benign fibroblastic tumor that most frequently affects males between the 5th and 7th decades of life. It presents as an asymptomatic mass usually in the subcutis, but fascial and muscular involvement can occur. It occurs most often in the soft tissues, but two cases have been described in the palate and one in the parotid gland. It is well-demarcated, white-grayish, firm and nodular. It is typically a paucicellular proliferation of spindle and stellate regular fibroblasts over a collagenous stroma that can be focally myxoid and can be SMA, CD-34 and keratin positive. Two cases studied showed a 2;11 (11q12) translocation. To the best of our knowledge, this is the first case described in the larynx. The case is presented to call attention that visceral cases of desmoplastic fibroblastoma can occur.

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PP3-247 LYMPHANGIOMATOUS POLYP OF THE PALATINE TONSIL: REPORT OF TWO CASES OF A RARE TONSILAR LESION Fred Ellinger, Fernando Siemann, Thiago Y. Miura, Karla S. Pereira, Adriana Brentegani, Luis C. Silva, Frederico G. N. Ellinger FAMEMA , Brazil BACKGROUND Benign tumors of the palatine tonsil are rarer than malignant ones. The most common is the squamous papilloma. Lymphangiomatous polyp of the tonsil is a rare lesion typically occurring in the palatine tonsil and considered a hamartoma. Until 2000, 36 cases have been plublished in the literature. We describe two cases to divulge this uncommon lesion. METHOD The two cases were seen recently in the Department of Pathology of the Faculty of Medecine of Marília. Hematoxylin and eosin–stained slides were reviewed. Clinical and follow-up information were obtained from the patients hospital charts. RESULTS Case reports: Clinical findings Case 1: P.M.R., fourteen years old white female with essential leukopenia and palmo-plantar hyperhydrosis presented to the department of othorhynolaryngology complaining of a sensation of a “meat ball” in the throat on deglutition, of two years duration. Examination showed a polyp in the left palatine tonsil. Bilatered tonsilectomy was performed with a diagnosis of left tonsilar polyp. The patient is currently asymptomatic. Case 2: E.W.S., nineteen years old mulatto male presented with a history of a foreign body sensation in the throat and choking, of two years duration. Examination found a polypoid mass in the right palatine tonsil. Bilateral tonsilectomy was performed and the postoperative course was uneventful. Pathologic findings Case 1 and 2: Macroscopy: In both cases a pedunculated tonsilar polyp was found. They were yellow-brownish, firm and measured 1,7 cm and 2,0 cm in greatest dimension in cases 1 and 2 respectively. Microscopy: histological appearances were similar in both cases. The pedunculated polyps were lined by non keratinizing squamous epithelium. The core of the polyps was composed of poorly cellular fibrous tissue containing numerous dilated vessels, mainly lymphatics. Underneath the epithelium, lymphoid tissue composed of small lymphocytes and some follicles, even with germinal centers, was focally found. There was focal epithelial lymphotropism with formation of small lymphocytic aggregates within the epithelium. Mature plasma cells were present in smaller numbers. Case 2 showed more abundant lymphoid tissue and larger clusters of lymphocytes within the epithelium than case 1. Pathologic diagnosis: Case 1 and 2: Lymphangiomatous polyp of palatine tonsil. CONCLUSION: Lymphangiomatous polyp of the tonsil is a rare benign hamartomatous lesion with typical location and histological appearance, here presented so that pathologists can learn to recognize and diagnose it. PP3-248 INVESTIGATION OF EPSTEIN-BARR VIRUS (EBV) PREVALENCE IN THYROID CARCINOMAS IN TUNISIA Mounir Trimeche, Sonia Ziadi, Mohamed Hachana, Khaled Amara, Badreddine Sriha, Moncef Mokni, Sadok Korbi Laboratory of Pathology, CHU Farhat Hached, Sousse, Tunisia Background: Epstein-Barr virus (EBV) infection has been linked with several malignancies of lymphoid and epithelial cell origin. It has been recently suggested that EBV may be involved in the progression of thyroid papillary carcinoma to undifferentiated carcinoma in Japanese patients. The purpose of our study was to evaluate the prevalence of EBV in thyroid carcinomas in Tunisia. Methods: To achieve this issue, 101 thyroid specimens from 101 patients, comprising malignant tumors (41 papillary carcinomas, 6 follicular carcinomas, 7 anaplastic carcinomas, 2 medullary carcinomas), and benign lesions (31 follicular adenomas, 10

multinodular goiters, 2 Hashimoto thyroiditis, and 2 Basedow disease), were randomly-selected. EBV infection was investigated by EBER1/2-in situ hybridization on formalin-fixed and paraffin-embedded tissues. Results: EBER1/2 was not detected in epithelial cells in any of the 101 specimens, but in 6 cases, EBV positivity was found in scattered normal lymphocytes. Conclusion: Given the sensitivity of the technique used, theses results do not support a relationship between EBV and thyroid carcinomas, in spite of EBV being high prevalent in our population when other tissue specimens are concerned. PP3-249 INVESTIGATION OF HUMAN PAPILLOMAVIRUS (HPV) IN BREAST CANCER AMONG WOMEN FROM TUNISIA Mounir Trimeche, Mohamed Hachana, Sonia Ziadi, Khaled Amara, Badreddine Sriha, Moncef Mokni, Sadok Korbi Laboratory of Pathology, CHU Farhat Hached, Sousse, Tunisia Introduction: Human papillomavirus (HPV) infection has been associated with a variety of benign and malignant epithelial lesions. The association of HPV with a subset of breast carcinomas has been reported but remains controversial. The aim of this study was to evaluate the frequency of HPV infection in breast carcinoma in Tunisia. Methods: 96 randomly selected invasive ductal carcinomas of the breast were investigated for the presence of HPV DNA sequences by polymerase chain reaction (PCR). DNA was extracted from frozen tumor tissues. HPV sequences were investigated by three different methods including the E1350L/E1547 primers amplifying a 180-bp sequence in a highly conserved region of a wide spectrum of anogenital HPV, the pU-1M/PU-2R primers specific for the oncogenic HPV types 16, 18, 31, 33, 52, 58, and the PU-31B/PU-2R primers for the low-risk HPV 6 and 11. Results: PCR for the detection of a 268bp sequence in the ȕ-globin gene demonstrated the presence of amplifiable DNA in all cases. No HPV-related DNA sequences were identified by PCR in any case of breast carcinomas tested. Discussion: Although our results may reflect a geographic feature, they do not support a relationship between the anogenital HPV tested and breast carcinomas; however, the possibility that other types of HPV contribute in the breast carcinogenesis can not be excluded. PP3-250 ALLELIC IMBALANCE OF THE SHORT ARM OF CHROMOSOME 3 IN NASOPHARYNGEAL CARCINOMA AMONG PATIENTS FROM TUNISIA Mounir Trimeche, Sonia Ziadi, Hend Braham, Mohamed Hachana, Khaled Amara, Moncef Mokni, Sadok Korbi Laboratory of Pathology, CHU Farhat Hached, Sousse, Tunisia Background: As part of North Africa, Tunisia is one of the world’s intermediate-risk areas for nasopharyngeal carcinoma. Loss of heterozygosity on the short arm of chromosome 3 (3p) is the most frequent genetic change reported in nasopharyngeal carcinoma from endemic areas. Method: We investigated the chromosome 3p for loss of heterozygosity in 47 microdissected primary nasopharyngeal carcinoma specimens and corresponding non-cancerous tissues from Tunisian patients using six microsatellite polymorphic markers. Results: Loss of heterozygosity was identified in 87.2% of cases on at least one of the six markers tested. The highest frequency of loss of heterozygosity was found at D3S1038 (60%), D3S1067 (57%), D3S1568 (52%), and D3S1076 (44%). Nine cases showed coexistence of loss of heterozygosity for three consecutive adjacent markers, suggesting a common deleted region flanked by D3S1568 (3p21.31) and D3S1067 (3p21.1-14.3), centered at D3S1076. Furthermore significant higher frequency of loss of heterozygosity were observed in patients older than 25 years than in patients under 25 years at the locus D3S1076 (57% versus 17%; p=0.02). Conclusion: These findings supports the hypotheses suggesting that loss of heterozygosity on chromosome

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3p in nasopharyngeal carcinoma is a common genetic event irrespective of geographic and ethnic origins, and suggests that one or more gene(s) affected by loss of heterozygosity at the locus D3S1076 were specifically involved in the pathogenesis of adult form of this cancer. PP3-251 CLINICOPATHOLOGIC STUDY OF CARCINOMA ASSOCIATED WITH SINONASAL INVERTED PAPILLOMA: NOT ASSOCIATED WITH EPSTEIN-BARR VIRUS Shih-Ming Jung1, Pao-Hsien Chu2, Chi-Ru Yeh1, Wen-Yu Chuang1, Ngan-Ming Tsang3 1 Department of Pathology, Chang Gung Memorial Hospital and Chang Gung Children Hospital, Chang Gung University College of Medicine, Taoyuan, Taiwan 2 Department of Internal Medicine, Taiwan 3 Department of Radiation Oncology , Taiwan Background: Carcinoma associated with sinonasal inverted papilloma is uncommon. Patients with inverted papillomas and carcinomas may occur in three patterns: 1) those who have carcinoma with recurrent inverted papilloma; 2) those whose tumors are predominate typical inverted papilloma with only focal invasive carcinoma or carcinoma in situ; 3) those who have carcinoma with a pattern similar to that of papilloma. Status of Epstein-Barr virus in the pathogenesis of inverted papilloma is controversial. We investigated the role of Epstein-Barr virus in carcinoma associated with sinonasal inverted papilloma. Method: Carcinoma associated with sinonasal inverted papilloma were retrieved from the surgical pathology files. Tumor DNA was extracted from paraffin blocks. Oligonucleotide primers for detecting LMP-1 gene were used to evaluate for the presence of Epstein-Barr virus DNA by a polymerase chain reaction (PCR) amplification. Results: Eleven cases were identified. Seven patients were female and four were male. The mean age was 52 (31-72) years. Two patients had moderately differentiated squamous cell carcinoma 162 and 148 months after excision of inverted papilloma with recurrences in the interim. Three cases presented inverted papilloma with focal invasive squamous cell carcinoma or carcinoma in situ. Six cases had carcinoma with a pattern similar to that of papilloma. Genomic DNA extraction was not workable in four of eleven cases. Epsten-Barr virus was not detected in the seven cases by PCR. Conclusions: The study demonstrates that the pathogenesis in carcinoma associated with sinonasal inverted papilloma is not dependent on concurrence of Epstein-Barr virus infection. PP3-252 SOLITARY FIBROUS TUMOR OF THE UVULA Vazquez Navarrete Sofia, Murga Tejada Carlos Hospital Del Sas La Linea, Spain Solitary fibrous tumor (SFT) is an uncommon spindle cell neoplasm rarely located in the oral cavity. We report a very unusual case of SFT located in the úvula.We have not found another one in this location in the literature. CASE REPORT A 80-years-old woman had a polipoid lesium 2 cms located in the úvula. The lesion was removed with a clinical diagnosis of Papiloma.Microscopically we found a spindle cell tumour with atipia , but no necrosis and some mitotic figures (< 4 in 10 HPF). The cells showed a strong immunoreactivity for vimentin and CD34, and variable for bcl-2 and negativity for other immunomarkers. The patient has been follow-up for 7 years (2000-2007)and in spite of a very small recurrence (may 2006) after removal of the new lesion, she is well and free of disease. Diagnosis of SFT extrathoracic is difficult and, although uncommon it should be considered in the differential diagnosis of oral soft tissue tumors.

PP3-253 HUMAN SALIVARY GLAND BRANCHING MORPHOGENESIS: MORPHOLOGICAL LOCALIZATION OF CLAUDINS AND ITS PARALLEL RELATION WITH DEVELOPMENTAL STAGES REVEALED BY EXPRESSION OF CYTOSKELETON AND SECRETION MARKERS Silvia Vanessa Lourenco1, Claudia Malheiros Coutinho-Camillo2, Marcilei Eliza Cavicchioli Buim2, Sabrina Hitomi Uyekita3, Fernando Augusto Soares4 1 Department of General Pathology, Dental School, University of São Paulo; Laboratory of Immunopathology, Institute of Tropical Medicine and Department of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil 2 Department of Pathology, Hospital A. C. Camargo, São Paulo, Brazil 3 Department of Dermatology, Medical School, University of São Paulo, São Paulo, Brazil 4 Department of General Pathology, Dental School, University of São Paulo; Department of Pathology, Hospital A. C. Camargo, São Paulo, Brazil Background: The development of salivary glands from simple precursor epithelial buds to functional glands is a highly complex and dynamic process termed branching morphogenesis, where tissue generate branched structures that subsequently differentiate into mature glands. Tight junctions (TJ) are thought to be the principal structures that contribute to cell polarity, by acting as an intramembrane barrier to prevent lateral movement of membrane proteins that form specific sites in the apical or basolateral membranes, thereby playing a critical role in the physiological functions of tubular organs. Several lines of evidence have demonstrated that claudins are directly involved in the formation of TJ strands as well as their barrier function. Method: Using immunohistochemistry and immunofluorescence we have mapped the distribution of claudins-1, 2, 3, 4, 5, 7 and 11 and compared it with the expression of differentiation markers in human salivary glands obtained from foetuses ranging from weeks 4 to 24 of gestation. Results: Expression of all claudins, except claudin-2 was detected in the various phases of human salivary gland development, up to fully mature salivary gland. The expression of all claudins increased according to the progression of salivary gland maturation evidenced by the classical markers – cytokeratin 14, cytokeratin low molecular weight, smooth muscle actin and human secretory component. Conclusion: Tight junction proteins – claudins appear to be important in the final shape and physiological functions of human salivary glands and are parallel related with markers of salivary gland differentiation. PP3-254 IMMUNOHISTOCHEMICAL STUDY OF TYPE I COLLAGEN EXPRESSION IN DIFFERENT DEGREES OF CARCINOMA EX- PLEOMORPHIC ADENOMA INVASION Vera Cavalcanti De Araújo1, Cristiane Furuse1, Patricia Ramos Cury1, Albina Altemani2, Venâncio Avancini Ferreira Alves3, Ney Soares De Araújo4 1 São Leopoldo Mandic Dental Research Center, Campinas, SP, Brazil 2 State University of Campinas School of Medicine, Campinas, SP, Brazil 3 University of São Paulo School of Medicine, São Paulo, SP, Brazil 4 University of São Paulo School of Dentistry, São Paulo, SP, Brazil Background: Carcinoma ex-pleomorphic adenoma (CXPA), a rare salivary gland tumour, is an important model to study the complex mechanism of tumour malignization and progression.

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Our group has already studied various aspects of this tumour concerning cells and stroma. Collagen is the major component of extracellular matrix proteins. Interaction between cells and collagen can regulate cellular gene expression, differentiation and growth. Besides it plays an important role in tumorigenicity. Recently, it was demonstrated that type I collagen induces disruption of E-cadherin-mediated cell-cell contacts and promotes cell proliferation. The aim of this study was to analyze the expression of type I collagen in CXPA. Method: Seventeen cases of CXPA, classified according to the degree of invasion in intracapsular, minimally invasive carcinoma and frankly invasive carcinoma with epithelial and/or myoepithelial component were immunostained for type I collagen. Results: Two patterns of type I collagen were disclosed: fibrilar; and amorphous or dense fibers. The fibrilar was interpreted as newly formed collagen and the amorphous or dense fibers as the mature collagen. The amorphous and dense fibers were observed in reminiscent pleomorphic adenoma stroma, in the capsule of the intracapsular type, in most of the stroma of intracapsular and invasive types of CXPA, and also as a dense line surrounding the glandular structures of the in situ areas. The fibrilar component was found in focal peripheral areas of the intracapsular and minimally invasive types, interpreted as invasion front. In frankly invasive type with only epithelial component the fibrilar pattern was found in focal areas involving small nests of cells and scattered throughout the stroma. When the tumour had myoepithelial cells as component, the fibrilar collagen, if present, was never in direct contact with tumoral cells. Conclusion: Fibrilar and dense fibers of type I collagen was observed in all types of CXPA. In CXPA with only epithelial component, focus of fibrilar collagen in peripheral border of the tumours and its presence surrounding and permeating small nests of cells in invasive type lead us to suggest that it should be an important component of the matrix in the invasion process of CXPA with epithelial component. Nevertheless, the same can not be postulated for the CXPA with myoepitelial component. PP3-255 HPV IN SITU HYBRIDIZATION ANALYSIS OF JUVENILE LARYNGEAL SQUAMOUS PAPILLOMATOSIS Samiah Zafar1, Herman Yee1, Youming Huang2, Hannah Wen3, Beverly Wang3 1 New York University School of Medicine, USA 2 Mount Sinai School of Medicine, USA 3 New York Univeristy School of Medicine, USA Background: Laryngeal squamous papillomas are benign neoplasms, often affecting the true vocal cords of children. They can be multiple with local recurrence, referred to as papillomatosis. Papillomas are thought to be associated with Human papilloma virus (HPV), demonstrated by using polymerase chain reaction (PCR). HPV in situ hybridization (HPV-ISH) is a novel technique that can be applied on paraffinembedded tissue with well-preserved histological architecture. We analyzed for low and high risk HPV types in the laryngeal papillomas by HPV-ISH. Materials and Methods: Twelve excisional biopsy specimens of the larynx were retrieved from 4 patients over a 4 year period (2003-2007) from Mount Sinai Medical Center. Age ranged from 4months to 9 years (average 5 years). All tissue was formalin-fixed and paraffin embedded. Automated HPV in situ hybridization (ISH) analysis was performed for the hybridization on 4μm thick tissue sections, using a Bench Mark XT (Ventana Medical Systems, Tucson, AZ) with DNA probes for low risk HPV (6,11) and high risk HPV (16,18,31,33,35,39,51,52,56,58, and 66). Positive and negative controls were hybridized alongside the study cases. Positive staining was visualized using the precipitating chromogenic reaction NBT/BCIP with a nuclear localization. Results: Laryngeal papillomas are soft excrescences grossly.

Histologically, they are composed of multiple slender, finger-like projections coved by squamous epithelium with central fibrovascular cores. In 1 patient (Case 1), focal mild cellular atypia was seen in a recurrent biopsy. No significant high-grade dysplasia seen in any of the biopsies. HPV-ISH revealed that 4 specimens from 2 patients (Cases 1 and 4) were positive for low risk HPV (50%). Three of these 4 specimens were recurrences from the same patient (Case 1). All twelve specimens of 4 patients were negative for high risk HPV (Table). Conclusions: Localized juvenile laryngeal papillomatosis is a benign condition related to HPV infection, with no evidence of high risk HPV. The identification of low risk HPV by ISH may be useful to predict local recurrence. HPV- ISH is a useful technique that can be easily performed on paraffin embedded tissue. PP3-256 SIGNET-RING CELL SALIVARY DUCT CARCINOMA A CASE REPORT Maria Papagianni1, Ioannis Dimitriadis1, Doxakis Anestakis1, Thomas Sidiras2, Frideriki Patakiouta3 1 Department of Pathology, Greece 2 Department of Otorhinolaryngology, Greece 3 Department of Pathology, Theagenion Cancer Hospital, Thessaloniki, Greece Background:Salivary duct carcinoma (SDC) was first described by Kleinsesser in 1968 and was classified as an independent entity by the 1991 Revised World Health Organization. It is a rare but highly malignant tumour. Purpose: We report a rare case of primary high-grade ductal carcinoma of salivary gland with signet-ring cells, that was confirmed by autopsy. Case report: A case of a high-grade ductal carcinoma of salivary gland originated “de novo” in parotid gland is presented. A 73 year-old male came to our hospital for a parotid swelling. He was treated by superficial parotidectomy. The tumour was unencapsulated, poorly circumscribed, grayish white, measuring 2,8cm in maximum diameter. The results of gross, microscopic, and immunohistochemical examinations are described. Microscopic examination showed that the tumour was entirely consisted of large signet-ring cells, positive for PAS and PAS-diastase. Lymphatic, vascular and nerve invasion were detected. Immunohistochemically, the tumour cells were positive for Ker7, Pankeratin, CEA, KerLMW and EMA, and negative for S-100 protein, KerHMW, PSAP, PSA, ER, PR, TTF1. Further ENTinvestigations including endoscopy and scans (MRT, ultrasound) showed no pathologic findings. In particular, growth of cervical lymph nodes and distant metastases were not revealed. Discussion: Salivary duct carcinoma is one of the most aggressive types of salivary gland carcinoma and is characterized by local invasion, lymphatic and hematogenous metastases and poor prognosis. The occurrence of signet-ring cells within salivary duct carcinoma is an extremely rare finding. The poor prognosis and aggressiveness of salivary duct carcinomas dictates aggressive therapy. PP3-257 RELATIONSHIP BETWEEN ACTINOMYCES AND CLINICOPATHOLOGIC FINDINGS IN ROUTINE TONSILLECTOMY AND ADENOIDECTOMY SPECIMENS Banu Dogan Gun, Burak Bahadir, Havva Gokce, Arzu Sonmez, Sukru Oguz Ozdamar Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, Zonguldak, Turkey BACKGROUND: Tonsillectomy and adenoidectomy are frequently performed operations but there is little information about the histological features. In this study it is aimed to investigate the presence of actinomyces in tonsillectomy and adenoidectomy specimens and analyse the patient’s profile and

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main pathological changes. METHOD: The medical records of patients who underwent tonsillectomy and adenoidectomy at Zonguldak Karaelmas University, Faculty of Medicine, Department of Pathology, in the period between January 2001 and December 2006 were included. RESULTS: Out of 387, 179 (46.25%) were female and 208 (53.75%) were male patients. Mean age was 12.69 years ranging from 2 to 67 years. There were 309 (79.85%) in the pediatric age group and 78 (20.15%) were adults. Main surgical indication was concomitant presence of recurrent throat infections and stenotic symptoms of upper airways. In 62 (16.03%) patients an adenoidectomy and tonsillectomy was performed, 248 (64.08%) underwent tonsillectomy alone and 77 (19.89%) adenoidectomy alone. There was actinomyces in 42 (10.85%) of the 387 patients, of whom 17 were adult and 25 were pediatric. That is, actinomyces was present in 17 of the 78 adult patients (21.7%), and 25 of the 309 pediatric patients (8.09%). It was almost always seen in tonsillectomy specimens, but in one of the adenoidectomy specimen. In adult population,; Actinomycosis was present in all of the obstructive patients (17/17), but none in tonsillitis cases (0/17); however in pediatric population actinomycosis ratio was nearly equal; 48% and 52%, in terms of lymphoid hyperplasia cases (12/25) and inflammatory conditions (13/25), subsequently. CONCLUSION: The presence of actinomycetes may indicate an etiologic role for this organism in adenoid and tonsillar inflammatory conditions and hypertrophy. Our results indicate that actinomycetes may play an active role in the etiology of hypertrophy of tonsillar disease in adult populations and as well as tonsillitis in pediatric patients as the causative agent. PP3-258 EXPRESSION OF Į-, ȕ-, AND y-CATENINS IN LARYNGEAL CARCINOMAS: CORRELATION WITH CLINICO-PATHOLOGICAL FEATURES Elena Collantes-Bellido1, Juan Ignacio De Diego2, Alejandra Caminoa1, Maria Del Pilar Prim2, Marta Mendiola1, BeléN San José3, Manuel Nistal1, David Hardisson1 1 Department of Pathology. Hospital Universitario La Paz. Universidad Autónoma de Madrid, Madrid, Spain 2 Department of Otorhinolaryngology. Hospital Universitario La Paz. Universidad Autónoma de Madrid, Madrid, Spain 3 Section of Biostatistics. Hospital Universitario La Paz. Universidad Autónoma de Madrid, Madrid, Spain BACKGROUND: Carcinoma of the larynx is one of the most frequent tumor in developed countries. Although several genetic and epigenetic alterations are characteristic of laryngeal carcinomas no definitive prognostic markers for advanced tumours have been identified. Catenins are a family of proteins including the Į-, ß-, and y-catenins. In the last years, it has been demonstrated that alterations in the cadherin-catenin adhesion complexes are involved in tumor initiation, progression and metastasis. The aim of this study was to evaluate the immunoreactivity of the Į-, ß-, and y-catenins in laryngeal squamous cell carcinomas (SCC) and to correlate their expression with clinicopathologic features. PATIENTS AND METHODS:This study involved a series of 279 formalin-fixed, paraffin-embedded laryngeal SCC. We constructed eight tissue microarray blocks where immunohistochemical staining was performed. Monoclonal antibodies against Į-catenin (clone Įcatenin-1, Dako, 1:35) ß-catenin (BD Biosciences, clone 14, 1:700), and y-catenin (BD Biosciences, clone 15, 1:1000) were applied using the Envision (Dako) method. The membranous expression of the catenins was scored as preserved, reduced or absent. RESULTS: Most of the tumors were located in the supraglottis (39.3%), followed by the glottis (25.5%), and subglottis (3%). The remaining cases were transglottic. Tumors were histologically classified as well (25%), moderately (42.6%) and poorly (32.4% differentiated. Overall, more than 75% of the tumors showed a reduced membranous expression of ȕ- and Į-

catenin, especially in poorly differentiated carcinomas. In contrast, only 31.1 % of the cases showed a reduced expression of y-catenin expression. Conserved membranous y-catenin expression was associated significantly with tumor size (T1 vs T4), location (glottis vs non-glottis) and differentiation (well differenciated vs poorly differentiated carcinomas) (p<0.005. No other associations between catenin expression patterns and recorded clinicopathological variables were detected. DISCUSSION: Several studies have suggested that loss of ȕcatenin protein is associated with tumour progression, metastasis and poor patient prognosis in many types of cancer, and this may be attributable to its role in cell adhesion. In this study, we have show that loss of expression of Į-, and ȕ-catenins is a common abnormality in SCC of the larynx. A preserved membranous pattern of y-catenin was found in glottic, well-differentiated and small tumors. However, expression of Į-, ß-, and y-catenins did not correlate with the prognosis of the patients. PP3-259 SECOND PRIMARY SQUAMOUS CELL CARCINOMA ARISING IN CUTANEOUS FLAP RECONSTRUCTIONS OF HEAD AND NECK CANCER PATIENTS: A RARE LONG-TERM COMPLICATION Yan Monnier1, Philippe Pache2, Philippe Monnier2, Snezana Andrejevic-Blant1 1 Institute of Pathology, CHUV-Hospital, Faculty of Biology and Medicine, University of Lausanne, 1011-Lausanne, Switzerland 2 Department of Otolaryngology and Head and Neck surgery, CHUV-Hospital, Faculty of Biology and Medicine, University of Lausanne, 1011-Lausanne, Switzerland The advances in reconstructive surgery have allowed surgical eradication of tumours which were considered inoperable in the past. Currently, surgical reconstructions by means of various types of skin flaps are commonly performed for advanced primary tumours of the head and neck. These techniques have been shown to increase loco-regional control of the disease and to improve functional rehabilitation. Early complications of regional and distant free flap transfers have been extensively described. However, there is still limited knowledge on the potential longterm complications of these techniques. Here, we report the cases of two patients with antecedents of squamous cell carcinoma of the head and neck (HNSCC), experiencing the development of a second primary SCC on the cutaneous surface of the flaps used for reconstruction. Growth of the second primary tumours occurred within a minimum of three year-interval. Both lesions arose at distance from the original mucosa of the oral cavity and from the deep surgical margins of the flaps. This observation ruled out any putative spread of a second primary tumour from the oral mucosa nearby the flaps. This is a rare complication of skin flaps reconstruction in HNSCC patients. Given the important expansion of these interventions in patients with head and neck cancers, this type of complication may become more frequent in the future and merit to be recognized by pathologists as a distinct entity. Interestingly, it seems that the well-known risk of a second primary HNSCC development in patients with previous head and neck carcinoma also applies to foreign tissues implanted in the area at risk. Therefore, long-term follow-up of patients previously treated for HNSCC, not only require careful evaluation of the normal mucosa of the upper aero-digestive tract but also of the cutaneous surface of the flap used for reconstruction.

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PP3-260 TWO DIFFERENT SALIVARY GLAND TUMOURS LOCATED IN PAROTIS GLAND AND PARAPHARYNGEAL REGION SYNCHRONOUSLY: A CASE REPORT Mehmet Kefeli1, Ilkser Akpolat1, Senem Cengel2, Filiz Karagoz1, Yucel Tanyeli2, Bedri Kandemir1 1 Ondokuz Mayis University, Faculty of Medicine, Department of Pathology ,Turkey 2 Ondokuz Mayis University, Faculty of Medicine, Department of Ear, Nose and Throat,Turkey BACKGROUND: The presence of more than one salivary gland tumour is a rare condition. Cytologic and histologic findings of a patient having two different tumours located in different regions (parotis gland and parapharyngeal area) at the same time are presented. CASE: A 58 years old male patient was admitted to the hospital with a mass in left side of the neck for 3 months. Physical examination revealed a mobile, painless, left jugulary mass (lymph node?) which was 4x3 cm in dimensions and revealed a parapharyngeal mass behind left tonsilla palatina. On the CT examination of cervical region and pharynx, two soft tissue masses are identified. The first mass was in the left parapharyngeal region and it was about 3,5x3x3 cm in dimensions, well-circumscribed and showed heterogeneous contrast uptake. The second one was 4,5x2,5x2 cm in dimensions, sharply delineated and homogeneous. It was located in the caudal region of the left parotid gland and partially embedded in to the gland. Initial clinical diagnosis was metastatic malign tumour. Fine needle aspiration was carried out for the cervical mass. Cytologic findings were consistent with Warthin tumour. The parapharyngeal mass was excised and microscopic examination diagnosis was pleomorphic adenoma. The servical mass was excised as partial parotidectomy 3 months later and microscopic examination diagnosis was Warthin tumour. When these two masses were evaluated with operative and clinical findings, they were determined unrelated anatomically. CONCLUSION: Although cytological examination is difficult in salivary gland lesions because of its complex morphology, fine needle aspiration of salivary gland lesions is a simple and cheap procedure which allows surgeon to decide the appropriate treatment before operation.

Soft Tissue, Joint and Bone PP3-261 SYNOVIAL SARCOMA: A RETROSPECTIVE ANALYSIS OF 12 CASES Amel Trabelsi, Sarra Mestiri, Ben Yacoub- Abid Lilia, Hanene Gasri, Med. Tahar Yacoubi, Soumaya Rammeh, Atef Ben Abdelkader, Sadok Korbi Pathology Department, F. Hached Hospital, 4000 Sousse, Tunisia Background: Synovial cell sarcoma is a mesenchymal malignancy that represents 10% of all soft tissues sarcomas. It most often occurs in the lower limbs of young adults. Our study aim is to report clinical and histopathological findings, treatment and outcome of such tumors. Methods: This is a retrospective study of 12 patients diagnosed with synovial sarcoma, retrieved from 1990 to 2005, at the pathology department of F. Hached hospital (Sousse, Tunisia). Results: The median age of patients was 21 years. The sex ratio was of 1. The most common sites of involvement were: The lower limbs (10 cases), the upper limbs (1 case) and the oropharynx (1 case). The revealing symptoms were: limb swelling in 11 cases, pain in 3 cases, dyspnea (1 case), cough (1 case) and a pulmonary metastasis (1 case). Our cases were classified according to the WHO’s classification in: monophasic type (8 cases), biphasic type (3 cases) and undifferentiated type (1 case). The tumors were classified in grade 2 (8 cases) and in grade 3 (4 cases), according the FNLCC recommandations. The treatment consisted in surgical removal in 10 cases (consisting in an amputation in 3 cases); adjuvant chemotherapy was indicated in 3 cases, a post-operative combination of chemo-radiotherapy was indicated in 1 case. Therapeutic abstention was indicated in 1 case. Another patient refused treatment. Clinical outcome was unfavourable with death occurrence in 6 cases. Conclusion: Synovial cell sarcoma carries usually a poor prognosis, that is correlated to multiple factors such as: anatomic location, size of tumor, histological type, grade, treatment modalities, especially the quality of surgical resection. PP3-262 GIANT CELL TUMOR OF BONE: A CLINICOPATHOLOGICAL STUDY 14 CASES Amel Trabelsi, Sarra Mestiri, Soumaya Rammeh, Fatma Kebir, Wided Stita, Lilia Ben Yacoub Abid, Med. Tahar Yacoubi, Sadok Korbi Pathology Department, F. Hached Hospital, 4000 Sousse, Tunisia Background: Giant cell tumor of bone are particular by their morphologic features and the absence of accurate prognostic parameters. Method: This is a retrospective study of 14 cases of giant cell tumor of bones diagnosed at the pathology department of F. Hached hospital in Sousse between 1987 and 2004. Radiographic and histopathological features were reviewed and confronted with both clinical data and patient’s follow-up. Results: 5 patients among six with a grade 1 tumor had a local recurrence; 1 patient among 8 with a grade 2 tumor had a local recurrence, another one developed pulmonary metastases after a local recurrence; 7 patients had neither recurrence nor metastasis after treatment. Conclusion: Histopathological parameters cannot be by their own predictive of a local recurrence or metastasis. Clinical and radiological features must be also taken in consideration.

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PP3-263 LETHAL COMPLICATIONS OF RHEUMATOID ARTHRITIS - A RETROSPECTIVE CLINICOPATHOLOGIC STUDY OF 161 AUTOPSY PATIENTS Miklós Bély1, Ágnes Apáthy2 1 Policlinic of the Hospitaller Brothers of St. John of God in Budapest, Hungary 2 National Institute of Rheumatology and Physiotherapy, Budapest, Hungary The aim of this study was to determine: (1) the spectrum and incidence of complications related to RA such as systemic vasculitis (SV) systemic AA amyloidosis (AAa), lethal septic infection (SI), carditis (endo-, myo-, epi-, or pancarditis), interstitial pneumonitis, interstitial lymphoid hyperplasia, polyserositis, glomerulonephritis, atlanto-axial subluxation, etc, (2) the mortality of complications related to RA, and (3) the clinically missed diagnosis of these. Patients and Methods A randomized autopsy population of 161 in-patients with RA was studied. RA was diagnosed clinically according to the ACR criteria. The complications of RA and cause of death were confirmed histologically. Results The lethal complications of RA were: Systemic vasculitis, which was found in 36 (22.4 %) of 161 RA patients, and led to death in 19 (11.8%) of 36 patients. SV was recognized clinically in 7 (19.4 rel%) of 36, and not recognized in 15 (78.95 rel%) of 19 patients with lethal complication of SV. AA amyloidosis complicated RA in 34 (21.1%) of 161 patients. AAa led to death by uremia in 17 (10.56%) of 34, and was clinically recognized in 9 (52.95 rel%) of 17 lethal cases, and not recognized in 8 (47.05 rel%) of 17 patients. Lethal septic infection was detected in 24 (14.9%) of 161 RA patients at autopsy, and was recognized clinically in 11 of 24 (45.8 rel%) cases. Cardiac insufficiency was the direct cause of death in 29 (18 %) of 161 patients, and was always recognized clinically. A rare form of interstitial pneumonitis (characterized by intensive interstitial lymphoid infiltration and secunder lymphoid follicle) led to death in 2 (1.24 %) of 161 cases, and was never diagnosed clinically. Interstitial nephritis in two (1.24 %), glomerulonephritis in one (0.62 %) of 161 RA patients led to death by uremia. The underlying disease leading to uremia was never recognized clinically. In 2 (0.9 %) of 161 patients acute liver necrosis (“yellow atrophy”), subacute liver necrosis (“red atrophy”) were the direct cause of death, clinically labeled as hepatorenal insufficiency but basically not recognized as the underlying disease leading to death. Discussion Two thirds of deaths were related to RA. Systemic vasculitis, AA amyloidosis and lethal septic infection were the most important major complications of RA. They were responsible for the death in 60 (53.1 rel%) of 113 cases related to RA (37.3 % of 161 RA patients). In this study systemic vasculitis was the most likely lethal complication to be missed clinically with high probability. PP3-264 PATHOANATOMIST'S VIEW ON CERTAIN PROBLEMS IN PRESENT-DAY ONCOLOGY Alexander Zubritsky Municipal Institution "Taldom Central Regional Hospital",Russia Mesothelioma is a comparatively rare neoplasm, more frequently affecting the pleura and peritoneum, less often - the pericardium. The diagnosis is imperfect, being usually delayed, and it is not by chance that this tumour remains an enigma for the oncologist. The accurate diagnosis is made at best while a pathomorphological examination of the removed tumour, at the worst - during a post-mortem examination.The author carried out a retrospective analysis of the pathoanatomical material at the Municipal Institution "Taldom Central Regional Hospital" (NCRH) during 1984-2004. Of the 1387 postmortem examinations of adults, we revealed only one case of malignant

diffuse highly differentiated papillary mesothelioma of predominantly the visceral peritoneum.Below is our case report.A 79-year-old woman was admitted to the Surgical Department of the TCRH on 6th February 1994, complaining of abdominal pain, and lack of defecation for two weeks, having considered herself sick during two months when she had developed lack of appetite and less of flesh. At examination by the physician on duty, the following diagnosis was made: partial ileus, predetermined, in all likelihood, by a colonic tumour. Surgery performed on 15th February 1994 was laparotomy: ovarian cancer with metastases into abdominal organs. The cytological study of ascitic fluid: adenocarcinoma. The woman died on day 8 after surgery. The final clinical diagnosis: ovarian cancer, involving abdominal organs;ascites;disseminated atherosclerosis.At post-mortem examination: the dead body of an undernourished elderly woman;the abdominal cavity contains up to 300 ml of reddish, muddy fluid.The greater omentum is deformed, solid, in the form of detached conglomerate. Pathoanatomical diagnosis:malignant diffuse mesothelioma predominantly of the visceral peritoneum with the histological form of papillary adenocarcinoma;acquired mechanical ileus; serous-fibrous peritonitis;generalized progressing atherosclerosis of the aorta, of the ileal arterias stage IV, degree 5 (according to Avtandilov). Hence peritoneal mesothelioma as the primary cause of death amounted to 0.072% amongst 1387 postmortem examinations of adult patients of our general somatic hospital over 21 years.In our opinion,one should reconsider the policy of regional oncologists towards improvement of the material and technical basis,activization of their activities and making courageous decisions aimed at using vitally important emergency chemotherapy in order to increase the survival rate. PP3-265 SUBCUTANEOUS SOLITARY FIBROUS TUMOUR OF THE BACK Barbara Paola Banzatti, Francesco Klinger, Valeria Bandi, Angelo Virgilio Pagliari, Marco Klinger Plastic Surgery Institute - University of Milan - Plastic Surgery Unit - Istituto Clinico Humanitas - IRCCS – Milano, Italy Background Wagner in 1870 is credited of the first histologic description of solitary fibrous tumour (SFT) an uncommon spindle cell neoplasm recognized as a distinct entity among primary pleural neoplasm by Klemperer in 1931. In the last years has been observed that SFT is anatomically ubiquitous and multiple new extrapleural locations of this rare mesenchymal neoplasm have been reported. SFT may be difficult to diagnose outside the usual locations because of its histologic variability and its back of distinctive features. Method A 42-year-old female presented with a slow-growing and painless subcutaneous mass in her back that was initially clinically suspected of being an epidermal cyst or a lipoma. The patient underwent simple excision under local anaesthesia of the tumour which was observed to be located in the deep subcutis and focally in close proximity of the fascia at tumour base without involvement of the underlying musculature. Results Histologically the excised tumour consisted of a proliferation of spindle cells arranged in a storiform pattern embedded in a fibrous matrix. A prominent vascular pattern, focal myxoid and thick hyalinized collagen fibers were observed. There were few normal mitotic figures, but no abnormal mitoses. The tumour cells were positive for CD34, bcl-2, CD99 and vimentin whereas stains for S-100, EMA, cytocheratin, desmin and alpha-smooth muscle actin were negative. Based on these findings a diagnosis of SFT was made. Conclusion Clinically SFT presents as a slow-growing mass in middle-aged adults with no sex predilection. The histogenesis of this tumour remains a matter of controversy. It has been recently suggested that SFT may represent the neoplastic transformation of bcl-2/CD34 fibroblastic-like dendritic cells involved in antigen presentation within the connective tissue. Grossly SFT are well

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demarcated, but partially encapsulated neoplasm. Histologically is characterized by a morphological appearance of alternating hypo- and hypercellular areas of spindle-shaped cells, dense bands of collagen and a vascular pattern. Differential diagnosis is sometimes difficult and is possible a mistaken with a variety of other soft tissue and spindle cell neoplasm including dermatofibrosarcoma protuberans, synovial sarcoma, schwannoma, spindle cell lipoma and cutaneous myofibroma. Immunohistochemistry is particularly important in the identification of rare and atypical soft tissue tumour. Immunohistochemically the diagnosis of SFT has been refined by the availability of immunohistochemical marker such as CD 34, bcl-2 and vimentin. PP3-266 COMPARISON OF TWO METHODS OF ABDOMINAL FAT PAD ASPIRATION AND BIOPSY IN DIAGNOSIS OF AMYLOID DEPOSITION IN PATIENTS WITH LONG STANDING RHEUMATOID ARTHRITIS Gholamhussein Alishiri1, Nasrin Shayanfar2, Roya Setareshenas2, Jafar Forghanizadeh3 1 Department of Rheumatology, Baqiyatallah University of Medical Sciences, Tehran, Iran 2 Department of Pathology, Iran university of Medical Sciences, Rasol akram Hospital Tehran, Iran 3 Department of Rheumatology, Iran university of Medical Sciences,Rasol Akram Hospital, Tehran, Iran Objective: To comparing two methods of abdominal fat pad aspiration and needle biopsy in diagnosis of secondary amyloidosis in patients with long standing rheumatoid arthritis (RA). Method: 220 consecutive patients (53 males and 167 females) with a history of at least 5 years of RA in two rheumatology clinics were studied. Abdominal subcutaneous fat pad aspiration (ASFA) and abdominal subcutaneous fat pad biopsy (ASFB) was performed. Specimens were stained by alkaline congo red method. All specimens examined for presence of apple green birefringence under polarized light. For confirmation, all positive biopsy specimens and 40 negative controls with long standing RA ( duration>15y) stained for AA protein by IHC method. Results: Amyloid was positive in 16 patients(7.3%). Nine of them were positive in both methods. Amyloid deposition was positive in 11(5%) patients with ASFA and in 15 (6.8%)patients with ASFB. All had 1+ deposits except in 2 of ASFB specimens, which had 2+ deposits. The sensitivity of ASFA and ASFB were 57% and 78.9% respectively. Conclusion: According to this study, the sensitivity of ASFA is lower than ASFB, however because ASFA is technically simple and less aggressive than ASFB, It is recommended that ASFA be used as first step in screening of patients with amyloidosis. PP3-267 HISTOPATHOLOGICAL ASSESSMENT OF THE EFFECT OF NON- ABSORBABLE PROSTHETIC MESH ON THE SPERMATIC CORD CONTENTS: AN EXPERIMENTAL STUDY Omran Zeinab1, Abbas Mohamed2, Nosseir Mona1 1 Theodor Bilharz Research Institute (TBRI), Department of Pathology, Egypt 2 Department of General Surgery Theodor Bilharz Research Institute (TBRI), Giza, Egypt Background: Previous studies have documented that the use of synthetic prosthesis in repair of inguinal hernia markedly reduces the incidence of recurrence. However, little is known about the effect of these prostheses on the spermatic cord contents. This work was carried out to study the histopathological changes in response to contact with the commonly used prostheses (Prolene & Mersilene) on spermatic cord contents in male dogs. Materials and Methods: Thirty two male dogs were included in this study.

The spermatic cords and testes were harvested from 2 dogs as a base line for the normal histology. The remaining 30 dogs were divided into 3 main groups. In the first (control) group (GI,n=10), the spermatic cord was delivered, its coverings were dissected and left intact on one side but on the other side it was skeletonized by removing its coverings. In the second group (GII,n=10), a strip of polypropylene (Prolene) mesh was wrapped snugly without strangulation around the dissected intact cord on one side and the skeletonized cord on the other side. A polyester (Mersilene) mesh was similarly applied in the third group (GIII,n=10). In each group, the spermatic cords and testes were examined histopathologically one week, and three months postoperatively. Results: At one week, the microscopic examination of the spermatic cord contents revealed more acute inflammatory reaction in mesh groups than the control. This reaction considerably subsided after three months with varying degrees and patterns of fibrous tissue deposition. These changes were more noticed in GIII than GII and in skeletonized cords than intact ones. The testes of all animals showed no histopathological changes. Conclusion: This experimental study proved the safety of the used non-absorbable prostheses on the spermatic cord contents and testes. Prolene mesh without skeletonization of the cord revealed more favorable results than Mersilene one. PP3-268 CYCLOOXYGENASE-2 EXPRESSION IN BENIGN AND MALIGNANT CHONDROID TUMORS Hye-Rim Park1, Yong-Koo Park2 1 Dept of Pathology, College of Medicine, Hallym University, Anyang, Korea 2 Dept of Pathology, College of Medicine, Kyung Hee University, Seoul, Korea Background: Recent studies have shown increased levels of cyclooxygenase-2 (COX-2) in various human malignancies to include various bone and soft tissue tumors. However, little is known with regard to COX-2 expression patterns in chondroid tumors. Method: Immunohistochemistry assays were performed for COX-2 in enchondromas (n=10), chondroblastomas (n=11), chondromyxoid fibromas (n=5), conventional chondrosarcomas (n=17), clear cell chondrosarcomas (n=7), and mesenchymal chondrosarcomas (n=6). Results: Among the benign chondroid tumors, chondroblastomas revealed characteristic strong positivity in 6 of 11 cases (54.5%). All enchondromas and chondromyxoid fibromas were negative except in one case. In conventional chondrosarcomas, three cases (17.6%) were strongly reactive with COX-2 and all positive cases represented grade III chondrosarcomas. Clear cell chondrosarcomas were found to be focally positive in two cases (28.5%), while all mesenchymal chondrosarcomas were negative. Conclusion: These findings suggest that COX-2 overexpression in conventional chondrosarcoma may represent both a negative prognostic factor and also an advanced histologic grade. Interestingly, expression of COX-2 in chondroblastomas could be an important factor for inducing peritumoral inflammatory changes in these specific tumors. PP3-269 EXPRESSION OF VASCULAR ENDOTHELIAL GROWTH FACTOR-C AND ITS RECEPTOR IN OSTEOSARCOMAS Hye-Rim Park1, Yong-Koo Park2 1 Dept of Pathology, College of Medicine, Hallym University, Anyang, Korea 2 Dept of Pathology, College of Medicine, Kyung Hee University, Seoul, Korea Background: Vascular endothelial growth factor-C (VEGF-C) and its receptor, vascular endothelial growth factor receptor-3 (VEGFR-3) have been implicated as important factors in the

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formation of lymphatic vessels, but its role in osteosarcomas has not yet been fully investigated. This study aims to define the expression of VEGF-C and VEGFR-3 in primary and metastatic osteosarcomas and their relationship to various clinicopathologic parameters. Method: Thirty three primary osteosarcomas and two pulmonary metastatic samples were immunostained for VEGF-C and VEGFR-3. In addition, VEGF-C and VEGF-D mRNA expression levels in three different human osteosarcoma cell lines and control fibroblasts were evaluated by real-time quantitative polymerase chain reaction (PCR). Results: Both VEGF-C and VEGFR-3 were mainly expressed in the cytoplasm of the tumor cells. Of the 35 patients with osteosarcoma, 16 patients (45.7%) showed strong positive reaction with VEGF-C. Four cases (11.4%) were negative and 15 cases (42.9%) showed weak immunoreactivity. For VEGFR-3, 12 patients ((34.3%) showed strong positive reaction. Fifteen cases (42.9%) were negative and 8 cases (22.8%) showed weak immunoreactivity. A significant positive correlation (Rs=0.42, p=0.01) was found between the expression of VEGF-C and VEGFR-3 in osteosarcomas. The expression of VEGF-C was significantly associated with osteoblastic subtype and high histologic grade in osteosarcomas. However, the expression of VEGF-C showed no significant correlation with the presence of metastasis. Expression of VEGFR-3 was not related to any clinicopathologic features analyzed. Two of the three tested osteosarcoma cell lines showed amplification of VEGF-C mRNA compared to control cells. No amplification of VEGF-D was noted in these cell lines. Conclusion: Our data suggest that VEGF-C and its receptor are expressed in osteosarcomas. Although the level of VEGF-C was high, it does not seem to have a direct influence on tumor metastasis in osteosarcomas. PP3-270 RETROPERITONEAL LYMPHANGIOLEIOMYOMATOSIS ARISING FROM CYSTIC ENDOMETRIOSIS Masaharu Fukunaga1, Aki Mistuda2, Makiko Fukunaga3, Kazutoshi Shibuya4 1 Department of Pathology, Jikei University Daisan Hospital, Japan 2 Department of Pathology, Toho Univeristy School of Medicine, Japan 3 Department of Pathology, Nihon University, School of Medicine, Japan 4 Deparment of Pathology, Toho University School of Medicine, Japan Aim and design: A case of retroperitoneal lymphangioleiomyomatosis (LAM) arising from cystic endometriosis was described. Results: A 25-year-old woman with no history of tuberous sclerosis or hormonal therapy presented with a painless, palpable abdominal mass. Computed tomographic and magnetic resonance imaging studies of the abdomen demonstrated a 4 cm cystic mass in the retroperitoneum. Macroscopically, the excised retroperitoneal cyst was multilocular and measured 4.0 X 3.5 X 3.5cm. Histologically, the lesion demonstrated three components. The first were multiple cysts or glands lined by columnar epithelial cells with cilia. The second component was a condensation of endometrial-type small stromal cells immediately subjacent to the cystic epithelium or glands. The third component was a thick exterior wall composed of plump spindle cells with clear to palely eosinophilic cytoplasm in a fascicular pattern, and slit-like vascular spaces, resembling LAM. Immunohistochemically, the epithelium and glands were positive for cytokeratin 7. The endometrial-type stroma cells were positive for vimentin and CD10. The cells of the LMA-like component showed positive staining for HMB45, alpha-smooth muscle actin, muscle actin and h-caldesmon. Conclusions: The lesion, LAM arising from endometriosis, represents a distinctive pathologic entity that

should be recognized and studied further. This type of lesion should be included in the differential diagnosis of retroperitoneal cystic lesions. PP3-271 ONCOGENIC OSTEOMALACIA ASSOCIATED WITH PHOSPHATURIC MESENCHYMAL TUMOR, A CASE REPORT AND REVIEW OF LITERATURE Maysa Al-Hussaini King Hussein Cancer Center, Jordan Oncogenic osteomalacia is an acquired syndrome characterized by renal phosphate wasting, hypophosphatemia, normal serum calcium levels and low serum 1,25-dihydroxyvitamin D osteomalacia caused by mesenchymal neoplasms that are termed phosphaturic mesenchymal tumors (PMTs). The condition improves or completely disappears after removal of the tumor. This is a case of a 22-year-old male who presented with a 1-year history of increasing bone pain and difficulty in walking. This was associated with generalised muscle weakness. Physical examination was unremarkable. Laboratory investigation revealed the presence of hypophosphatemia, phosphaturia, elevated alkaline phosphatase levels in the presence of normal calcium and parathyroid hormone levels. By MRI a paraspinal heterogenous well circumscribed mass was seen.Complete excision of the mass was performed. By microscopic examination, phosphaturic mesenchymal tumor was diagnosed. Oncogenic osteomalacia as well as phosphaturic mesenchymal tumor are disscused along with review of the literature. PP3-272 GIANT CELL BIPHASIC SYNOVIAL SARCOMA WITH MASSIVE OSTEOID AND BONE FORMATION SIMULATING OSTEOSARCOMA Lankar Abdelaziz1, Laouar Omar2, Oukid Nadia1, Belghersa Lamine1, Kassouri Med Lamine3, Bouzitouna Mahdjoub3, Maza Rabeh3 1 Department of Pathology University Hospital Annaba-Algeria 2 LAOUAR-Algeria 3 Department of Orthopedical Surgery University Hospital Constantine-Algeria Introduction Synovial sarcoma with giant cells and predominant osteoïd and bone formation is a rare variant difficult to distinguish from osteosarcoma especially in presence of lytic areas. Case report We report a case in a man aged 49 years admitted in Orthopaedic Department for pathologic fracture of the upper right femoral metaphysis. Radiography shows a wellcircumscribed and calcified tumour of 37mm diameter with lytic areas in upper metaphysis and diaphysis of the right femur. The surgeon discovers a hard, reddish, rounded mass with focal calcifications occupying the external hillside of the fracture and infiltrating the neighboring bone. Histopathologic examination leads to a diagnosis of giant cell biphasic synovial sarcoma with massive osteoïd and bone formation. The pattern is biphasic with epithelial component expressing intensely CK and EMA, numerous mononuclear and giant cells and extensive osteoid and bone formation. Discussion Diagnosis is difficult because of the existence of lytic areas accompanying the tumor, massive osteoïd and bone formation and the character of the tumour cells associating a mononuclear background and giant multinucleated cells. It is important for the pathologist not to misdiagnosis this rare entity of synovial sarcoma because of its different course from osteosarcoma. The main argument which plead in favour of synovial sarcoma is essentially biphasic pattern with glandular and squamous component expressing CK and EMA.

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PP3-273 ALVEOLAR SOFT PART SARCOMA OF BONE, A CASE REPORT Fariba Abbasi, Ahmad Reza Afshar Urmia University of Medical Sciences, Iran Background:Alveolar soft part sarcoma (ASPS) is a distinct sarcoma that was defined by Christopherson et al in 1952.The incidence of ASPS has been reported to be less than 1% of all the primary soft tissue malignancies.Primary involvement of bone by ASPS is extremely rare. ASPS usually grows as a painless mass without functional impairement.Grossly,the tumor is well circumscribed,usually firm,grayish- yellow.Microscopically, the tumor cells are separated by fibrovascular tissue into nests.The cells are large and have vesicular, prominent nuclei and a granular cytoplasm.Mitoses are exceptional.Intracytoplasmic PAS positive diastase resistant crystalline material is diagnostic of the tumor. Case presentation:We report a 20 year old man with swelling of left heel.Radiography showed a well- defined large lytic lesion of calcaneous bone.Microscopic examination of curettage derived sample showed proliferation of large polygonal cells with vesicular nuclei and eosinophilic cytoplasm arranged in nests.Intracytoplasmic needle like PAS positive structures were seen.Immunohistochemical staining for S-100 protein was negative.Staining for desmin and vimentin was focally positive. Results:The mentioned microscopic findings confirmed the diagnosis of ASPS..Three months after curettage of the lesion metastatic foci were revealed in lung. Conclusion:Primary involvement of bone by ASPS is extremely rare,however it should be considered in differential diagnosis of primary lytic bone lesions with distant metastases,in particular metastatic hypernephroma in a young patient. PP3-274 CLEAR CELL ADENOCARCINOMA OF ABDOMINAL WALL Thouraya Achach, Soumaya Rammeh, Amel Trabelsi, Lilia Ben Yacoub-Abid, Wided Stita, Fatima Zahra Kebir, Mounir Trimech, Moncef Mokni, Sadok Korbi Department of Pathology. Hospital Farhat Hached, Sousse, Tunisia Background Endometriosis is a frequent benign disorder. Several observations of the coexistence of endometriosis and cancer have been published. Malignancy arising in extra-ovarian endometriosis is a rare event. Method Through a new case of clear cell adenocarcinoma associated with endometriosis foci occurring in the abdominal wall of a 49-year-old woman, we highlist the possibility of endometriosis malignant transformation and we review recommended criteria for this diagnosis. Results The patient was operated 20-years before for uterus leiomyoma. Actually, she presented with a large painful abdominal mass. After radiologic exploration, she was operated with a diagnosis of desmoid tumor. Histological examination showed a clear cell adenocarcinoma associated to endometriosis foci. Exploration of uterus and ovary did not show any malignancy or endometriosis. Conclusion When confronted to an extraovarian tumor with endometrial appearances, pathologists should concider a primary tumor and an origin in endometriosis. The demonstration of endometriosis might require the examination of multiple sections. Confirmation that a tumor is of endometrial type is of clinical importance. PP3-275 MALIGNANT PERIPHERAL NERVE SHEATH TUMOUR ARISING IN ASSOCIATION WITH ECTOMESENCHYMOMA: DOES IT REALLY EXIST? Guillermo Muñoz1, Francesco Felipo1, Isabel Marquina1, Mar Pascual1, Ana Fuertes1, Celia Del Agua2, Luis Plaza1 1 Pathology Department, Miguel Servet University Hospital, Zaragoza, Spain 2 Pathology Department, Alcañiz Hospital, Alcañiz(Teruel),Spain

BACKGROUND: Two situations have been described in which a MPNST is associated with ganglioneuromatous or ganglioneuroblastomatous tissue; the first, when one of the tissues transforms into a MPNST; the second, when the ganglioneuromatous portion of an ectomesenchymoma does the same. Ectomesenchymoma is a tumour that usually appears during paediatric age, and it is composed by rabdommyoblastic elements, mature ganglion cells and neuroma-like structures. MATERIAL AND METHODS: Newborn male that presents a right paratesticular mass of 4x3x1,5 cm, wich is removed. RESULTS: Histological study reveals three well differentiated areas; one has a nodular pattern, and it is composed by small round blue cells with Homer-Wright rosettes, reactive for neuroendocrine and skeletal muscle immunohistochemical markers. Other areas are fusiform and loose, and present abundant scattered mature ganglion cells. It is also seen densely cellular areas consisting of spindle atypical cells with a high mitotic index, reactive for S-100 protein. It is diagnosed as malignant ectomesenchymoma with areas of high grade MPNST, originated from the schwannian constituent of ganglioneuroblastoma. CONCLUSION: Does the possibility of a MPNST developing from the ganglioneuromatous portion of an ectomesenchymoma actually exist? Two cases of complex tumours which contain neuroblasts or ganglion cells and also mesenchymal elements, usually rhabdomyosarcoma have been described, although there are authors who are uncertain about them based on the fact that, in spite having MPNST, neither of them reports clear images of it. Besides, Conzutto et al. believe that an ectomesenchymoma must not have features of MPNST. Our case shows evident areas of rhabdomyosarcoma and ganglioneuroblastoma, with focus of MPNST, which leads us to consider it an ectomesenchymoma that undergoes transformation to MPNST. PP3-276 PRIMARY OSSEOUS RHABDOMYOSARCOMA WITH FOCAL MATRIX FORMATION MIMICKING OSTEOSARCOMA: A CASE REPORT Radzislaw Kordek1, Piotr Sowa1, Dorota Jesionek-Kupnicka1, Michaá Panasiuk2, Marek Kmieciak2, Cezary Chudobinski3 1 Department of Oncology, Medical University Lodz 2 Department of Orthopaedic Surgery, Copernicus Memorial Hospital Lodz, Poland 3 Department of Radiology, Copernicus Memorial Hospital Lodz , Poland We present an unusual case of primary osseous pleomorphic rhabdomyosarcoma with focal matrix formation mimicking osteosarcoma. The patient was a 21-year-old man who suffered for two months with pain and slight enlargement of his left calf. The plain film demonstrates large, predominantly osteolytic mass in the region of proximal fibula with typical features of malignant primary tumour. On open surgical biopsy, the tumour was consisted of atypical cells, some presenting spindle morphology. Between them, there were bands of densely hyalinized matrix with osteoid appearance, but without definite lacunae or calcifications, and the diagnosis of osteosarcoma was established. Consequently, the tumor was removed. The tissue presented more pleomorphic cells with some definite rhabdomyoblasts. Desmin, actin, Myf4 and MyoD1 were positive in tumor cells and the diagnosis of rhabdomyosarcoma was eventually established. A few cases of primary pure rhabdomyosarcoma of bone were reported, and there were also reported cases of other bone tumours with rhabdomyosarcomatous differentiation: dedifferentiated chondrosarcoma, fibrosarcoma and osteosarcoma. Our case does nor meet criteria for sclerosing rhabdomyosarcoma as matrix formation is focal and cells are spindle and pleomorphic, but it shows the another possiblity for mistake with primary osseous tumour.

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PP3-277 CORRELATION BETWEEN MICROVASCULAR DENSITY AND DIFFERENTIATION GRADE IN SOFT TISSUE TUMOURS Ovidiu Mederle1, Narcisa Mederle2, Dragos Andrei Izvernariu1, Marius Raica1 1 University of Medicine and Pharmacy, Timisoara, Romania 2 Veterinary Medicine Faculty, Timisoara, Romania Introduction Although there are thousands of publications on the subject of tumour angiogenesis, their great majority refer to carcinomas, and the publications on the subject in soft tissue tumours are very rare and, until present, with discordant results. According to this perspective, we have made the assessment of microvascular density in soft tissue tumours. Materials and methods 54 cases of soft tissue tumours (43 malignant and 11 benign tumours) were studied according to the following protocol: surgical samples were fixed in buffered formalin, embedded in paraffin, sectioned at 5 ȝ, then stained for morphological diagnosis (H&E), and for immunohistochemical detection of microvessels (smooth muscle actin – SMA, CD31 and CD34). Weidnet method of microvessel counting was used. Results All the tumours included in the study have presented rich blood vessel network, the majority (>80%) of the blood vessels from within the tumour area being of immature and intermediate type (positive for the endothelial markers and negative for SMA). This aspect was more obvious in the case of liposarcoma and malignant fibrous histiocytoma, but with no correlation with the differentiation degree. The highest values of microvascular density (MVD) were recorded in liposarcoma and malignant fibrous histiocytoma, in straight correlation with the high number of immature and intermediate blood vessels. We have noticed major differences in the blood vessel number between benign and malignant tumours, which were statistically significant (p<0.02). We have also noticed significant differences between the group composed of leiomyosarcomas and fibrosarcomas on one side, and liposarcoma and malignant fibrous histiocytoma on the other side (p<0.05). The results of microvascular density assessments were concordant for either endothelial marker used (table). Microvascular density was significantly higher in the malignant tumours than in the benign ones. Tumour MVD/CD34 MVD/CD31 Benign tumours 10.3 9.4 Fibrosarcoma 34.6 32.2 Liposarcoma 61.2 58.3 Malignant fibrous histiocytoma 58.9 55.7 Leiomyosarcoma 21.5 22.4 Conclusions Malignant soft tissue tumours have a rich blood vessel network, especially the liposarcoma and malignant fibrous histiocytoma. The majority of the blood vessels from within the tumour area are of immature and intermediate type. MVD does not correlate with the differentiation degree, but have significantly higher values in malignant tumours then in the benign ones. PP3-278 FLOW CYTOMETRIC ANALYSIS OF DNA PLOIDY AND S-PHASE FRACTION IN PRIMARY LOCALIZED MYXOFIBROSARCOMA: CORRELATIONS WITH CLINICOPATHOLOGICAL FACTORS, Skp2 EXPRESSION, AND PATIENT SURVIVAL Chien-Feng Li1, Wen-Wei Huang2, Hock-Liew Eng3, Ching-Nan Lin1, Shih-Cheng Chou4, Shih-Chen Yu3, Hsuan-Ying Huang3 1 Department of Pathology, Chi-Mei Foundation Medical Center, Tainan, Taiwan 2 Department of Family Medicine, Buddhist Dalin Tzu Chi General Hospital, Chiayi, Taiwan 3 Department of Pathology, Chang Gung Memorial HospitalKaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan 4 Department of Pathology and Laboratory Medicine, Veterans General Hospital-Kaohsiung, Kaohsiung, Taiwan

Background: Histological assessment for prognostication of myxofibrosarcomas remains challenging, for which we recently identified Skp2, an oncoprotein involved in S-phase progression, as an independent prognosticator . Methods: We assessed S-phase fraction (SPF) and DNA ploidy of myxofibrosarcomas and the association between SPF and Skp2. Flow cytometric findings were analyzed by CellQuest software for 75 primary myxofibrosarcomas and correlated with clinicopathological factors, Skp2 labeling index (LI), metastasis-free survival (MeFS), and overall survival (OS). Results: Forty-eight and 27 cases were classified as diploid and non-diploid, respectively. High SPF (҈20%) was detected in 32 of 61 interpretable cases. Skp2 overexpression (LI ҈ 10%) was seen in 36 of 72 cases with successful scoring. Non-diploidy was associated with higher FNCLCC grades (p=0.006), remarkable necrosis (p=0.010), and Skp2 overexpression (p=0.018). Noticeably, SPF was significantly related to Skp2 LI (p<0.001, r=0.458) and other clinicopathological factors, including FNCLCC grade, AJCC stage, and mitotic rate, etc. Non-diploidy predicted shorter OS (p=0.0045) and MeFS (p=0.0489), whereas SPF ҈ 20% was associated with inferior MeFS (p=0.0252) alone. In multivariate analyses, non-diploidy remained as an independent predictor for both OS (p=0.020, RR=3.337) and MeFS (p=0.013, RR=5.780), together with Skp2 overexpression (p=0.014 for OS; p=0.017 for MeFS) and positive margins (p=0.004 for OS; p=0.002 for MeFS). Conclusion: Skp2 overexpression promotes S-phase progression in myxofibrosarcomas, with a strong association between Skp2 LI and SPF. However, SPF, probably overshadowed by Skp2, provides no independent prognostic utility. Non-diploidy, along with Skp2 overexpression and positive margins, is independently predictive of adverse outcomes. PP3-279 INTRAMUSCULAR HEMANGIOMA. REPORT OF 41 CASES Samir Amr, Salwa Sheikh Dhahran Health Center, Dhahran, Saudi Arabia Intramuscular hemangiomas are uncommon soft tissue tumors with potential for recurrence. There are two large series in the literature (Allen and Enzinger: 89 cases; and Beham and Fletcher: 74 cases). Several single case reports or series of few cases are reported. We report herein a series of 41 cases. During 30-year period (1974-2003), a total of 41 cases of intramuscular hemangiomas were diagnosed at our laboratory. There were 16 males and 25 females. The mean age at presentation was 20 years with a range from 1 to 62 years. 15 patients (38%) were less than 15 years, and 33 (80%) were less than 30 years of age. The tumors were located in the upper limbs (10 cases), lower limbs (13 cases), trunk (8 cases) and head and neck (10 cases). The tumors were classified according to the type of blood vessels dominating the lesion. In some instances, a mixture of histologic patterns was observed in the same case. The commonest variant was the cavernous type (22 cases). Other variants included capillary (6), venous (6) and mixed (7). Several tumors showed the presence of thrombi (9) or phleboliths (10). Occasional tumors showed Masson lesion (Intravascular papillary endothelial hyperplasia). Most lesions showed variable amount of fatty tissue. Four tumors recurred locally, most probably due to incomplete excision on initial surgery.

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PP3-280 CANINE OSTEOSARCOMA: HISTOGENETIC AND CELL PROLIFERATION CHARACTERISTICS Josemara Cavalcanti, Eliane Amstalden UNICAMP-State University of Campinas, SP, Brazil Osteosarcoma is the most common of all malignant canine bone tumors and has a high aggressiveness potential. From the histological point of view it is a tumor with a heterogeneous cell pattern that is characterized by the production of bone matrix and/or immature bone by atypical osteoblasts. Systematic studies with a objective to evaluate histogenetic aspects and the cellular proliferation index in canine osteosarcomas have not yet been described. The objective of this study was to determine the differentiation of the cellular elements and evaluate the cell proliferation index of these tumors correlating them with their different morphological subtypes, using markers such as: vimentin, osteocalcin, S-100 protein, 1A4, HHF53, factor VIII, AE1/AE3, CD31, CD34 and Ki-67. A retrospective study was performed using 65 cases of canine osteosarcoma, clinically evaluated according to sex, breed, age and topography of the lesions. Tumors were classified according to their histological subtype in: nonproductive, moderately productive and productive osteoblastic tumors, chondroblastic, fibroblastic, giant cell type, telangiectatic and undifferentiated subtypes. Males from the Rottweiler breed with an average age of 7,4 years were more frequently affected by osteosarcoma. In 77,4% of cases, the lesions were on the appendicular skeleton (mainly on femur). The identified histological subtypes were: osteoblastic (47,7%), fibroblastic (23,1%), chondroblastic (21,5 %), giant cell type (4,7%), telangiectatic (1,5%) and undifferentiated(1,5%).In 98,2% of all tumors there was vimentin reactivity. Immunoreactivity to osteocalcin occurred in osteoblastic, chondroblastic and fibroblastic cells of 91% of all osteosarcomas. The S-100 protein was expressed in 79,7% of the cases (100% of the chondroblastic subtype). A significant proportion of osteosarcomas expressed positively to 1A4 and HHF35 antibodies. All osteosarcomas were negative for AE1/AE3 and CD31 and CD34 testing. Ki-67 analysis revealed a high cell proliferation index in all cases nonproductive osteoblastic osteossarcoma subtype. Canine osteosarcomas are constituted by a diverse cell population, composed of osteoblastic, chondroblastic, myofibroblastic and mioblastic elements, which are probably originated from a single pluripotent mesenchymal cell or from immature osteoblasts that go thru a differentiation process. Nonproductive osteoblastic osteosarcoma presented a considerably high cell proliferation index that may relate to a more aggressive behavior. PP3-281 FIBROMATOSIS-PRIMARY AND FIRST RECURRENT TUMORS. A FLOW CYTOMETRIC AND CLINICAL ANALYSIS IN SEARCH TO PREDICTORS OF OUTCOME Eliane Amstalden1, Frederic Preffer2, Andrew Rosenberg2 1 State University of Campinas, SP, Brazil 2 Massachusetts General Hospital, Boston, MA, USA Fibromatosis is a benign fibrous tissue tumor that has a significant propensity to locally recur. To help identify features predictive of aggressive behavior, we analyzed retrospectively, a large series (68 cases) of deep seated and plantar fibromatosis. We determined the phases of cell cycle and DNA concentration, recorded clinical information, and evaluated a variety of histological features (degree of cellularity, mitotic index, stroma composition, margin of growth, and status of resection margin) of primary (50 cases) and first recurrent (18 cases) tumors. We correlated these findings with the patient's outcome to ascertain prognostic parameters. The patients included thirty-eighty males and 30 females who ranged in age from 5-79 (mean 34) years. All

tumors were resected and nineteen patients also received radiotherapy. Recurrences developed in 38 patients. Tumor size ranged from 1 to 16 (mean= 6.7) cm. Flow cytometric analysis was performed on 51 tumors (35 primary and 16 first recurrent tumors). All tumors were diploid and greater than 93% of the analyzed cells were in the G0/G1 (resting) phase. The morphologic and flow cytometric findings in fibromatosis were similar in the primary and first recurrent tumors and were not associated with any prognostic features. However patient age, involvement of surgical margins and tumor size were predictive of local recurrence. PP3-282 AGGRESIVE ANGIOMYXOMA. A REPORT OF TWO CASES IN MEN Alva MartíNez-Angoa, Mario Alfonso Reséndiz-Morán, Ernesto Carrera-González, Mercedez Hernández-González Hospital General de México, Mexico These are the first two cases of aggresive angiomyxoma diagnosed and treated in men in the General Hospital of Mexico City. The first case was a 69-years-old man, presented with an inguinal mass, at first diagnosed as an hernia. The second case was a 32-year-old man with an increased volume in the scrotum diagnosed as lymphedema. The aggresive angiomyxoma (AA) is a soft tissue neoplasm found in pelvis or perineum, locally aggressive but not metastatic, it is frequently found in women, but it is very rare in men. However, it seems to have a better prognosis in men. The tumors in both men and women must be differentiated from intramuscular myxoma, myxoid neurofibroma and angiomyofibroblastoma, all of them benign neoplasms without any risk of relapse or local invasion. In several cases, the AA can be mistaken with a malignancy, because of the size and infiltration of surrounding tissues. So it is important to recognize it to select the correct treatment and give a prognosis. PP3-283 REACTIVE ANGIOENDOTHELIOMATOSIS IN A PATIENT WITH CHRONIC RENAL FAILURE George Zimmer1, Alex Ochsner2, Herbert Koppl1, Urs Wagner1, Hans-Martin Riehle1, Christian Bussmann1, Markus Zuber2, Carole Gengler3, Louis Guillou3 1 Viollier Histopathologie, Basel, Switzerland 2 Departement Chirurgie, Chirurgische Klinik, Kantonsspital Olten, Switzerland 3 Institut Universitaire de Pathologie, Lausanne, Switzerland BACKGROUND: Reactive angioendotheliomatosis (RAE) is a rare condition of unknown pathogenesis characterized by a reactive cutaneous vascular proliferation that often occurs in the setting of coexistent systemic disease. Different terms have been used for this lesion, including systemic proliferating angioendotheliomatosis, diffuse dermal angiomatosis and glomeruloid angiomatosis. METHODS: We describe a case of RAE in a patient with chronic renal failure. RESULTS: A 66year-old woman presented with a dermal swelling of the left thigh. No previous trauma was recorded. The patient had several concomitant diseases, including diabetic chronic renal failure, anaemia, hypertension, coronary heart disease, mitral and aortic valve disease. Calcium serum level was normal. Histologically, the lesion was characterized by a diffuse proliferation of anastomosing capillaries within the dermis and subcutis. Endothelial cells showed focal hobnailing, but lacked significant atypia or significant mitotic activity. Media calcinosis was observed in some dermal arteries as well as foci of subcutaneous fat necrosis. Immunhistochemically, endothelial cells expressed CD31 and CD34 and were negative for HHV8. Smooth muscle actin stained pericytes. CONCLUSION: RAE is a rare, probably under-recognized benign lesion, which can easily be confused with other benign and malignant vascular neoplasia. RAE is

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commonly associated with underlying systemic diseases, including renal or hepatic failure, chronic infections, rheumatoid arthritis, valvular heart disease, and cryoglobulinemia. Immunoreactivity for HHV8 has been detected in up to 40% of the cases. PP3-284 DISTRIBUTION OF CHONDROCYTES CONTAINING ALPHA-SMOOTH MUSCLE ACTIN IN HUMAN NORMAL, OSTEOARTHROTIC AND TRANSPLANTATED ARTICULAR CARTILAGE Ctibor Povysil Institute of Pathology, 1st Medical Faculty and University General Hospital, Prague, Czech Republic Background: Relatively little is known about the expression of a contractile actin isoform, alpha smooth muscle actin, in the normal human articular chondrocytes and in chondrocytes during certain pathological conditions. The aim of our work was to evaluate the occurence of alpha-smooth muscle actin in normal, osteoarthrotic and transplantated articular cartilage by means of immunohistochemistry. Method: Histology and immunohistochemistry were performed on paraffin embedded cartilage specimens obtained during total hip replacement for femoral neck fracture (15cases) and or osteoarthrosis (15 cases). Moreover, tibial plateau were retrieved from seven males with haemophilic arthropathy undergoing total knee arthroplasty. We also used samples from 9 patients with autologous chondrocytes transplantastion in the region of the traumatic defect in the distal femur 10 months after operation. Results: Approximately 20% of the chondrocytes in the superficial region of normal cartilage expressed alpha-smooth muscle actin as demonstrated by immunohistochemistry. It was readily apparent that fever cells in deep region contained alpha-smooth muscle actin. In the osteoarthrotic cartilage and cartilage from haemophilic patients occurred predominantly cells comprising clonal grouping stained for alpha-smooth muscle actin. In regions with formation of fibrocartilagineous repair tissue the majority of cells stained also positive. In the case of transplantated autologous chondrocytes the samples obtained 10 months after operations new cartilage had partly hyaline character and the majority of cartilage cells contained alpha-smooth muscle actin. Conclusion: Our study showed that significant percentage of articular chondrocytes express alpha-smooth muscle actin in health, disease , response to injury, defect healing and after autologous transplantation. They may utilize the contractile actin isoform in manipulating the extracellular matrix of articular cartilage. PP3-285 CUTANEOUS LEIOMYOSARCOMA Anastasia Nikolaidou1, Christos Makos2, Elisavet Pazarli1, Theodoros Balampanidis3, Rodi Kotakidou4, Symela Amanatidou5 1 Department of Pathology, General Hospital of Kilkis, Greece 2 Department of Oral and Maxillofacial Surgery, General Hospital of Kilkis, Greece 3 Department of Otorinolaryngology Head and Neck Surgery, General Hospital of Kilkis, Greece 4 Department of Pathology, General Hospital ‘’G. Gennimatas’’, Thessaloniki, Greece 5 Technician, Greece BACKGROUND: Superficial leiomyosarcomas are rare soft tissue tumors that account for less than 3% of all cutaneous soft tissue neoplasms. They further divided into cutaneous and subcutaneous and they usually occur in the extremities. METHOD AND RESULTS: We report a case of an 80-year-old female patient who came for excision of a firm protruding nodule at the temporal area, measuring 3,5 cm. Histological examination of the specimens of the resected nodule showed a spindle-cell

tumor with morphologic and immunohistochemical features compatible with leiomyosarcoma; the tumor was positive for smooth-muscle actin, caldesmon, desmin and vimentin but negative for S-100 protein, HMB-45 and pankeratin. CONCLUSION: Immunohistochemical study seems necessary due to the difficulties in the differential diagnosis, which includes spindle-cell dermal carcinoma, spindle-cell melanoma and malignant fibrous histiocytoma. Furthermore the location of this particular neoplasm is considered very rare. PP3-286 GLOMANGIOMA OF THE THIGH: CASE REPORT. CLINICOPATHOLOGICAL AND IMMUNOHISTOCHEMICAL FINDINGS Lydia Abou-Asabeh, George Sfyroeras, Konstantinos Moulakakis, Nikolaos Bessias, Calypso Barbatis Hellenic Red Cross Hospital, Greece Background: Glomus tumors are usually benign perivascular neoplasms composed of cells resembling modified smooth muscle cells of the normal glomus body with three variants: the solid type, glomangioma and glomangiomyoma. They are sporadic but autosomal dominant inheritance with linkage to chromosome 1p21-22 (locus VGLOM), leading to mutation of the glomulin gene has been reported. The thigh is an exceptionally rare site with less than 10 published cases. Methods: A 41-year-old female presented with a disturbing, painless mass at the inner side of her right thigh. An MRI scan revealed a mass fed by a branch of the deep femoral artery situated in between the adductor muscles of the right thigh. The tumor was excised and it was an encapsulated, 3.5 cm, brown, friable mass with recognisable transversing fibrous bands. The tissue was formalin fixed and examined with H+E, PAS, PASdiastase and with the immunohistochemical method of EnvisionHRP for detection of vimentin, Į-SMA, desmin, CD 34, factor VIII, CD 99, Bcl 2, c-kit, chromogranin, mixed cytokeratins, ki 67, S-100 and p 53. Results: The tumor was an encapsulated, non invasive mass with a complete fibrous vascular capsule, surrounded by fibrofatty tissue with scattered striated muscle fibers. The bulk of the tumor mass comprised large vascular channels with thin or hyalinised wall and in between the cellular areas were composed of oval cells with uniform hyperchromatic nuclei (ki 67 <1%,) and eosinophilic cytoplasm arranged in compact, trabecular masses, single cells or as perivascular wreath like. The neoplastic cells were vimentin, Į-SMA, CD 34, Bcl 2 positive with focal strong expression of desmin. With PASdiastase a distinct perivascular linear positivity was noted and a histological diagnosis of glomangioma was made. Conclusion: This is a case of a well circumscribed glomangioma of the right thigh, with an aberrant immunophenotype in an unusual site which required differential diagnosis from hemangioma, cellular leiomyoma and neuroendocrine tumors. PP3-287 SOFT TISSUE SARCOMAS: REVIEW OF 614 CASES Pelin Bagci1, Nuray Kepil1, Hale Demir1, Sergulen Dervisoglu1, Didem Colpan2, Fazilet Dincbas2, Nil Molinas Mandel3, Murat Hiz4, Pembe Cagatay5 1 Istanbul University Cerrahpasa Medical Faculty,Department of Pathology, Istanbul, Turkey 2 Istanbul University Cerrahpasa Medical Faculty,Department of Radiation Oncology, Istanbul, Turkey 3 Istanbul University Cerrahpasa Medical Faculty, Department of Medical Oncology, Istanbul, Turkey 4 Istanbul University Cerrahpasa Medical Faculty,Department of Orthopaedic Surgery, Istanbul, Turkey 5 Istanbul University Cerrahpasa Medical Faculty,Department of Biostatistics, Istanbul, Turkey

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BACKGROUND: Even with the reform in molecular genetics , soft tissue sarcomas (STS) are still among the main problems of diagnostic pathology and the pathogenesis of most soft tissue tumors is still unknown. We need larger epidemiologic studies to define the pathogenesis and to find out the most effective therapy options. METHOD: We reviewed 614 cases diagnosed in our department between the years 2000- 2007 according to the latest WHO classification; and analyzed them considering age, sex, etiology, localization, recurrence, metastasis, and therapy. RESULTS: Fifty-five percent of the cases were between 21-60 years; and the median age was 46 years. In the first two decades of life trunk and abdomen and then lower extremities were the most popular localizations. There were no significant relation between age and sex. Immunhistochemistry was used for the tumors in early ages (<40), but histological analyses were enough in adult tumors. Tumors of uncertain differantiation group were mostly diagnosed type (21%). Highest recurrence rate was seen in abdominal tumors and especially in liposarcomas. Lung was the first site for metastasis and the highest metastasis rate was seen in synovial sarcoma. There were only 2 postradiotherapy sarcomas; a synovial and a pleomorphic sarcoma. The prefered therapy option was mostly surgery; but chemotherapy and radiotherapy were also used for the aproppriate tumors. CONCLUSION: The median age in our material is younger then in the English literature. Distribution of localization is the same; but there is no correlation between gender and other variables. Tumors of uncertain differantiation constitute the largest part of our review. As we cannot obtain the origin of many sarcomas even with immunhistochemistry, histopathologic examination is still the gold standart in sarcoma diagnosis. PP3-288 REPORT OF THREE CASES OF MESENTERIC FIBROMATOSIS. IS IMMUNOHISTOCHEMISTRY NECESSARY FOR THE DIAGNOSIS? Hariclia Kalekou1, Ioannis Efstratiou2, Stavroula Pervana2, Ioannis Skandalos3 1 Pathology Department, Saint Paul General Hospital, Greece 2 Papageorgiou General Hospital Institute of Pathology, Greece 3 Surgical Department, Saint Paul General Hospital, Greece Mesenteric fibromatosis is a form of deep fibromatosis. According to the 2002 WHO classification fibromatoses belong to the category of intermediate , locally aggressive tumors which are characterized by infiltrative growth and local recurrences but no metastatic potential. Both sporadic cases and cases associated with familial adenomatous polyposis have been described. The purpose of this study is to report the histologic and immunohistochemical findings in 3 cases of mesenteric fibromatosis and discuss the role of immunohistochemistry in the diagnosis of these tumors. Two of our patients were female, aged 64 and 29 years and one male, aged 33. One gave a history of previous abdominal surgery, one presented with abdominal discomfort six months postpartum and the third presented with acute abdomen. None had familial adenomatous polyposis. In the first case the tumor extended from the mesentery to the pancreas infiltrating large vessels and was only partially excised. In the other two cases the tumors involved the mesentery, infiltrated the wall of the small and the large bowel respectively and were excised together with a segment of the involved bowel. All three cases exhibited a proliferation of spindle cells with myofibroblastic features forming long fascicles and associated with collagenous and focally loose stroma. Despite the variable cellularity and the infiltrative border, mitoses were absent to rare and there was no atypia or pleomorphism. Inflammatory infiltrates were distinctively absent. The immunohistochemical evaluation of the tumors included staining for Keratin AE1/AE3, CD34, S100 protein, smooth muscle actin, desmin, CD117 which were all negative and for ȕ-catenin which was positive with characteristic nuclear staining. The diagnosis of mesenteric

fibromatosis usually relies on careful observation of the histologic features in H+E sections. The immunohistochemical evaluation is a helpful adjunct in cases in which there is doubt regarding its distinction from sclerosing mesenteritis and gastrointestinal stromal tumor(GIST) or as part of protocol immunohistochemistry in spindle cell lesions of the mesentery. Our finding of nuclear ȕ-catenin positivity in the three cases studied is consistent with previous studies that have shown ȕcatenin positivity in mesenteric fibromatosis as compared to the negativity in GIST and in sclerosing mesenteritis. PP3-289 MORPHOLOGICAL AND CLINICAL CHARACTERISTICS OF THE PERIODONTAL TISSUES INFLAMMATION RESULTED FROM THE JAW FRACTURES Volodymyr Gavrilov, Sergiy Morozov Lugansk State Medical University, Lugansk, Ukraine Numerous epidemiological data suggested that periodontal tissues inflammation resulted from the conservative therapy of jaw fractures have high incidence rate in Ukraine. The aim of present study was to find out the morphological and clinical markers of the effectiveness of the improved design of a dental splint device combined with the ‘Uimisbol’ gel &Bolus Alba treatment in patients with jaw fractures. Methods. 25 volunteers were enrolled while receiving dental care for the jaw fractures. Each patient had a fixing of the jaw by the improved Tigershtedtt’s splints. The gel “Uymisbol” &Bolus Alba treatment was used locally on the fracture area in the form of the oral applications. The main active substance in “Uymisbol” was a plant extract of the Comarum palustre. 10 patients were treated in routine way without ‘Uimisbol’ gel &Bolus Alba application (group of comparison). Histological slides of gum tissue biopcies stained with H&E, Van Gison, and then stereological evaluation by point count method performed. The volume fractions of inflammatory infiltrates, vascularity, areas of dystrophy and necrosis of connecting tissue and others were determined. The results showed in the group of comparison the increase of volume fractions of inflammatory infiltrates, vessels, foci of periodontal connective tissue dystrophy and necrosis. The conservative therapy with the gel “Uymisbol” resulted in the reduction of hyperemia and vessels number. Inflammatory infiltrates and areas of connecting tissue dystrophy revealed occasionally. The improved device did not damage periodontal tissues in aesthetically important areas of oral cavity Conclusion These findings suggested the jaw fractions without improved treatment stimulate persistent inflammation in the gum of patients. On this background there is braking of repair processes, which can unfavorably alter bone repair processes. The use of gel “Uymisbol” &Bolus Alba treatment, possessing antioxidant, osteotropic, anti inflammatory and immune stimulating properties allows normalizing gum tissue’s metabolism, to recover the normal parameters of gum tissues and accelerate bone repair. PP3-290 CYTOGENETIC STUDY OF 108 ADIPOSE TISSUE TUMORS Piera Balzarini1, Paola Dal Cin2, Andrea Tironi1, Elisa Rossi1, Mariella Chiudinelli1, Paolo Pandolfi3, Maria Flocchini3, Piergiovanni Grigolato1 1 2nd Department of Pathology, University of Brescia, Spedali Civili, Brescia 2 Cytogenetic Laboratory, Brigham and Women's Hospital, Boston 3 Plastic Surgery Unit, Spedali Civili, Brescia Background. Cytogenetic investigation of soft tissue tumours, revealing clonal chromosome alterations associated to specific gene sites, allows better comprehension of pathogenesis and more

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precise classification of soft tissue neoplasms. We studied 108 adipose tissue tumours from 105 patients, 63 males, 42 females, mean age 52 (range 23-79). Tumour sites were mainly shoulder, forehead, dorsum, arm and cervical region. Size of tumours was ranging between 0.5 cm and 10.5 cm. Methods. All the 108 adipose tissue tumours were cytogenetically analysed. Chromosome preparations were performed after short-term culture, using standard cytogenetic technique and karyotype were established according to ISCN nomenclature. Hystopathologic features and karyotype of tumours were investigated and reported independently. Results. According to pathological diagnosis, 73 tumours were conventional lipoma, 12 fibrolipoma, 10 angiolipoma, 7 spindle cell lipoma, 1 lipomatosis, 4 atypical lipomatous tumours/well differentiated liposarcoma (ALT/WDL), 1 de-differentiated liposarcoma. Cytogenetically, 51,4% of lipomas showed abnormal karyotype involving 12q13-15 (58,3%), other sequences of chromosome 12, mainly 12p11-12 and 12q24 (22,2%) and chromosome 6 (16,7%). Angiolipomas showed a normal diploid karyotype in 90% of cases. Spindle cell/pleomorphic lipoma showed monosomy of chromosome 13 and 16 in 42,9% of cases, translocation of 6p21-23 with Xp21.1, 1p32 and 11q13 in 42,9% of cases and balanced translocation of chromosome 12 in 28,6% of cases. Five liposarcomas, 4 ALT/WDL and 1 de-differentiated liposarcoma, showed a complex karyotype, characterized by ring- and giantsupernumerary chromosomes. One case of ALT/WDL was a recurrence and the morphology was deceptively benign in despite of the presence of ring- and giant-chromosome. Conclusion. In the 108 cases of adipose tissue tumours analised cytogenetic studies confirm differences between lipoma and liposarcoma. Although the same chromosome can be involved in benign and malignant neoplasms, aberrations are quite different. The present case-study shows that in conventional lipoma chromosome 12 frequently translocates with 3q27-28, while in liposarcoma chromosome 12 gives rise to ring- and giant- chromosomes. So far even when lipomatous tumour has minimal atypical morphology, cytogenetic analysis still shows it’s sarcomatous nature. Although histopathological examination is the gold standard in lipomatous tumour diagnosis, cytogenetic analysis can help in difficult cases. PP3-291 PRIMARY EMBRYONAL RHABDOMYOSARCOMA OF LONG BONE IN A CHILD 5 YEARS OLD. A CASE REPORT AND REVIEW OF THE LITERATURE Catherine Michail-Strantzia, Irene Bonou-Boukouvalea, Panagiota Giamarelou, Chris Zambakidis, Helen Kosmidis Children's Hospital of Athens "P. & A. Kyriakou", Greece Background: RMS is a well documented soft tissue sarcoma. The majority of RMS that involve bone in children are metastatic or represent secondary involvement by contiguous growth of a soft tissue lesion. This case is the sixth one involving the femur in childhood. Design: We present the Clinico-Radio-Pathological findings in a boy 5 years old, who submitted with a painful left knee’s joint due to a left limb swelling. Results: The x-ray, CTscan and MRI of the left femur revealed a diaphyseal large lytic lesion with cortical destruction and reactive periostic “onion skinlike” proliferation, compatible with a Ewing’s Sarcoma. The histological and immunohistochemical investigation evaluated a high grade spindle cell sarcoma, with a herringbone pattern of proliferation, enlarged, bizarre or polygonal cells and abundant mitosis without any producing of malignant osteoid. The diagnosis of RMS has been prevailed over the diagnosis of Ewing’s Sarcoma and supported by the positive reaction in Desmin, Vimentin, Myf-4 and MyO-D1 abs. The patient has been treated according to the RMS protocol and five months later the bone tumor has been completely resected showing histologically 95% tumor necrosis and two nodules of live RMS without any evidence of malignant osteoid production. Conclusion: We

ordinarily do not make the diagnosis of a RMS in a bone tumor. Although the clinical and radiological data are compatible with those of a Ewing’s Sarcoma, the immunohistochemical results support the diagnosis of a RMS involving the periosteum and the adjacent skeletal mass. RMS described in children’s bones have not specific radiographic features and show bone destruction by a lytic process. Histologically they do not differ from RMS of soft tissue and any type of them can occur in bone. The presence of other component, such as cartilage, bone or undifferentiated sarcomatous elements in bones raise the suspicion that the lesion in question may represent dedifferentiation phenomenon rather than a primary RMS of bone or may be a part of a malignant mesenchymoma.

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Cardiovascular Pathology PP3-292 SYSTEMIC AND CORONARY VASCULITIS IN RHEUMATOID ARTHRITIS - A RETROSPECTIVE CLINICOPATHOLOGIC STUDY OF 161 AUTOPSY PATIENTS Miklós Bély1, Ágnes Apáthy2 1 Policlinic of the Hospitaller Brothers of St. John of God in Budapest, Hungary 2 National Institute of Rheumatology and Physiotherapy, Budapest, Hungary Systemic vasculitis (SV) is one of the main, and the most likely lethal complication to be missed clinically with high probability of rheumatoid arthritis (RA). The aim of this study was to determine: (a) the prevalence, the mortality, and the clinically missed diagnosis of SV at autopsy in RA, (b) the prevalence, the mortality, and the clinically missed diagnosis of coronary arteritis and arteriolitis (CAa). Patients and Methods A non-selected autopsy population of 161 in-patients with rheumatoid arthritis was studied. RA was confirmed clinically according to the criteria of the ACR. Vasculitis was diagnosed histologically. Results (a) SV was found in 36 (22.4%) of 161 RA patients. SV directly led to death in 19 (11.8%) of 36 patients (due to coronary arteritis and/or arteriolitis in 12, pulmonary and bronchial arteriolitis with disseminated lobular-sublobular pneumonia in 3, cerebral vasculitis and multifocal brain necrosis in 2, thrombovasculitis of renal artery and renal necrosis in 1, or thrombovasculitis of mesenteric artery and hemorrhagic intestinal necrosis in 1 patient). SV was recognized clinically in 7 of 19 with the lethal complication of SV. (b) Coronary arteritis and/or arteriolitis were associated with SV in 25 of 36 cases (15.5 %). CAa led directly to death in 12 of 25 patients (due to coronary arteritis and thrombosis of the main coronary artery with a large myocardial infarct in 1, coronary arteriolitis and multiple focal microinfarctions of the myocardium (myocardiocytolysis) in 11 cases). Coronary arteritis and/or arteriolitis was detected clinically only in 3 of 25 patients. Discussion In RA various organs are involved by vasculitis with different incidence. The leading and most endangered target is the heart, which was involved in this study in two thirds of 36 RA patients with SV. In most cases the arterioles and the small arteries are affected by vasculitis. Three types of vasculitis (non-specific, fibrinoid necrotic, granulomatous) may be present simultaneously in different vessels or combined in one. Different stages of inflammation can be found simultaneously, reflecting histologically the relapsing nature of this type of vasculitis. In most of the cases the clinical diagnosis of vasculitis is based on the visible skin involvement. Vasculitis of the skin does not necessarily run parallel with the heart involvement. Coronary vasculitis should be kept in mind, and it is very important to look for minor, transient, and recurrent clinical signs of cardiac disease, especially if there is a history of vasculitis. PP3-293 PATHOGENESIS SILENT MYOCARDIAL INFARCTION IN RHEUMATOID ARTHRITIS - A RETROSPECTIVE CLINICOPATHOLOGIC STUDY OF 161 AUTOPSY PATIENTS Miklós Bély1, Ágnes Apáthy2 1 Policlinic of the Hospitaller Brothers of St. John of God 2 National Institute of Rheumatology and Physiotherapy, Budapest, Hungary Coronary vasculitis is the main cause of silent myocardial infarction in rheumatoid arthritis (RA), and the most likely lethal complication to be missed clinically with high probability. The aim of this study was to outline the pathogenesis of silent

myocardial infarction due to coronary arteritis and/or coronary arteriolitis (CAa) in RA. Methods A non-selected autopsy population of 161 in-patients with RA was studied. RA was confirmed clinically and CAa was diagnosed histologically. Results 1. SV was found in 36 (22.36%) of 161 RA patients. CAa was associated with SV in 25 of 36 cases (15.5 %, 69.44 rel%). Subepicardial (main) coronary arteritis with, or without intramural coronary arteriolitis were present in 14 (56 rel%) of 25 patients. Coronary arteritis and thrombosis of the main coronary artery with a large myocardial infarct led directly to death in 1 (4 rel%) of 25 patients Coronary arteriolitis with multiple focal microinfarctions of the myocardium (myocardiocytolysis) was the direct cause of death in 11 (44 rel%) of 25 cases. 2. Three types of vasculitis (non-specific, fibrinoid necrotic, granulomatous) were present simultaneously in different vessels or combined in one. Different (acut-subacute-subchronic-chronic) stages of inflammation can be found simultaneously. Discussion All types of autoimmune vasculitis are of a relapsing (recurrent) nature. Histologically different stages of inflammation can be found simultaneously, reflecting histologically the relapsing nature of vasculitis. Vasculitis of the main coronary arteries with or without thrombosis may result in ischemia and may lead to a large myocardial infarct, macroscopically similar to myocardial necrosis due to coronary atherosclerosis and/or thrombosis. Vasculitis of the arterioles and small arteries can lead to local ischaemia and to regressive changes. This process is more or less widespread and multifocal, depending on the number of involved vessels. The necrotic foci are small and they are present in different stages of necrobiosis; small (1-2 mm of diameter) homogeneous areas alternating with small lytic foci of myocardium, and scars of a similar size simultaneously side by side. Because of the recurrent nature of immune vasculitis the regressive changes accumulate with time in the myocardium and may lead to unexpected sudden death. Clinically it is difficult to recognize the small accumulating myocardial necrosis (myocardiocytolysis). The history of vasculitis, transient cardiac complaints, low voltage electrocardiogram (ECG) may help in the diagnosis. PP3-294 PATHOBIOLOGY OF ATHEROGENESIS Vasileos Anestiadis1, Vladimir Nagornev2 , Eremei Zota3, Ilie Tsiple1, Vasile V. Anestiadi1 1 Centre for Pathobiology and Pathology, Academy of Sciences, Chisinau, Moldova 2 Research Institute of Experimental Medicine, Academy of Medical Sciences, Saint-Petersburg, Russia 3 Medical University, Chisinau, Moldova Background. We have studied the role of the immune inflammation (IINF) in the set-up and development of atherosclerosis, both in experimental material and in samples of human coronary arteries, obtained during surgical operations, from 250 patients. Methods. The material has been studied by ultrastructural and immunohistochemical methods. Results. The role of the lymphocytes in the atherogenesis (AG) is viewed from the standpoint of their input into the expression of the lymphocytes and monokines, into the regulation of the production of chemokines and in situ antibody synthesis. The input of the smooth muscle cells (SMC) into the AG is analyzed not only from the standpoint of the proliferation of the synthetic phenotype of the SMC and the synthesis of connective tissue matrix proteins, but also taking into account their involvement into IINF reactions, expression of cytokines and class II antigens. During initial AG the depositing and/or formation into the subendothelial intima layer of modified LDL, which acquired autoantigenic properties, is the triggering factor, which stimulates the expression by endothelial cells of chemoadhesive molecules, which connect with the ligands of non-granular leucocytes. Activated monocytes/macrophages and T-cells, which penetrate

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into intima, even during the most early stages of the AG, start a cascade of reactions, that makes sure the development of the early phase of the inflammation. Conclusion. The involvement of the intimal cells into the inflammatory reaction (macrophages, SMC and endothelial cells begin to produce class II antigens) transfers the acute phase of the inflammation into a chronic one, like the delayed hypersensitivity reaction. PP3-295 TWO AUTOPSY CASES OF A NEW CORONARY DISEASE (ISOLATED EOSINOPHILIC CORONARY PERIARTERITIS) Kajihara Hiroki1, Tachiyama Yoshiro2, Hayashi Yuzo3 1 Div. Path., Dept. Health Sci., Prefectural University of Hiroshima, Japan 2 Div. Path., Hiroshima Nishi Medical Center, Japan 3 Div. Path., Hiroshima City Asa Hospital, Japan Ist case: A 40y male presented symptoms of angina pectoris for 9 years and expired with symptoms of unstable angina. At the time of admission, 26 days before death, blood examination showed WBC5,000/mm3(St 3%, Seg 38%, Ly 47%, Eo 4%, Ba 5% and Mo 3%). At the autopsy, the heart weighed 430g and both right and left coronary arteries of the subepicardial region were grayish white in color and elastic hard. Histologically, inflammatory infiltration was localized only in adventitia of coronary arteries located in the subepicardial region. Inflammatory cells infiltrated in the adventitia were mostly eosinophils. The medial smooth muscle cells were slightly hypertrophied and the intima showed irregular thickening with fibrosis. Inflammatory change could not be found, not only in the intramural coronary arteries, but also in the arteries of other tissue and organs. IInd case: A 28y female suffered from bronchial asthma for 4 years and showed symptoms of angina pectoris about 3 months before death. Blood examination showed WBC8,600/mm3 with slight eosinophilia. At the autopsy, the heart weighed 220g and epicardial coronary arteries were grayish white in color, the same as case I. Histologically, inflammatory cells, mainly of eosinophils, infiltrated also in the adventitia of both right and left epicardial coronary arteries. The medial smooth muscle cells were well preserved, but the intima of these arteries showed irregular thickening with fibrosis. No findings of angiitis could be detected in the blood vessels except subepicardial coronary arteries. As a result of these autopsy findings described above, this type of coronary arteritis could be termed "Isolated Eosinophilic Coronary Periarteritis", a new coronary disease. PP3-296 RENOVASCULAR HYPERTENSION CAUSED BY GIANT-CELL ARTERITIS – CASE REPORT Vesna Mihajlovic-Bozic1, Svetislav Tatic2 1 Department of Pathology, Institute for Cardiovascular Diseases, Clinical Center of Serbia, Belgrade, Serbia 2 Institute of Pathology, Belgrade University School of Medicine, Serbia Background: Giant-cell arteritis (GCA) affects almost exclusively persons older than 50 years of age, and disease risk is highest among those who are 75 to 85 years of age. Two thirds of those affected are women. Giant cell arteritis typically causes vasculitis of the extracranial branches of the aorta and spares intracranial vessels. Transmural inflammation of the arteries induces luminal occlusion through intimal hyperplasia. Clinical symptoms reflect end-organ ischemia. We reported a case of a 15-year-old male teenager with severe heart and acute renal failure as the dominant clinical manifesations of renovascular hypertension (RVH) caused by atypical (GCA). Unrecognized RVH and treatment of the consequent heart failure by angiotensin-converting enzyme inhibitors (ACEI) probably contributed to progression of renovascular disease to bilateral renal artery occlusion. Contrast

angiography of the abdominal aorta and renal arteries disclosed bilateral renal artery occlusion and the patient was reffered for surgical revascularization. The diagnosis of giant-cell arteritis is established by biopsy. Material and methods: Surgical specimen is a part of the wall of the right renal artery. The material was fixed in formalin, filtered and embedded in paraffin. Serial sections of the renal artery were stained:H&E, Masson trichrome and Elastic Van Gieson. Microscopic finding of the resected right renal artery consistent with giant cell arteritis: Numerous multinucleated giant cells mainly close to the internal elastic lamina surrounded by nonspecific lymphocyte infiltrates. The elastic lamina is partially fragmented or thickened. The lumen is partially occluded by intimal hyperplasia with superimposed organized thrombus. Disscusion: Fibromuscular dysplasia is the most common cause of RVH in children and GCA is an exceptionally rare cause of RVH. Primary or isolated involvement of renal arteries is more, often described in Takayashu arteritis. The onset of symptoms in GCA may be acute or insidious. Conclusion: Unrecognized RVH treated with ACEI probably provoked acute renal failure and contributed to recurrent heart failure and “flash” pulmonary edema in our teenage patient with atypical GCA. Prompt normalization of heart function and arterial hypertension occured after adequate therapy which is related with GCA. PP3-297 BONE FORMATION IN CARDIAC VALVES Ivo Steiner Department of Pathology, Charles University Faculty of Medicine and Faculty Hospital, Hradec Kralove, Czech Republic Background: Dystrophic calcification is the most common pathological finding in operatively excised cardiac valves. Several reports describe bone in calcified cardiac valves (heterotopic ossification; bony metaplasia) and recent studies have identified bone proteins in such valves. Method: Histological and immunohistological examination of 1177 calcified cardiac valves operatively excised at the Department of Cardiac Surgery. Results: There were 128 patients (10.9 %) with bone and/or cartilage formation in the excised valve (119 aortic, 9 mitral). The incidence was higher in men (12.8 %) than in women (8.5 %). The average age of patients with metaplasia (64.1 years) did not differ from the average age of those without it (64.3 years). It was higher in females (66.9 years) than in males (62.6 years). Metaplastic changes were encountered in all types of calcific aortic valve disease (senile, bicuspid valve, postrheumatic). The marrow of the heterotopic mature lamellar bone was usually formed by adipose tissue. There was often chronic inflammation with an infiltrate composed predominantly of polyclonal plasma cells. Rarely, complete hematopoiesis was present. Conclusion: Heterotopic bone formation is closely related to the process of calcification, such that its presence has been described particularly in the aortic valve, in the calcified annulus of the mitral valve and, rarely, in arterial atherosclerotic plaques. In the past, calcific aortic disease was thought to be due to “degenerative”, time-dependent, wear-and-tear of the leaflets with passive dystrophic calcium deposition. Now, there is compelling data to suggest that calcific aortic disease is an active disease process similar to atherosclerosis. A crucial role in the process of calcific aortic stenosis is played by valvular interstitial (fibroblast-like) cells that, under various stimuli, become activated to myofibroblast-like cells. A subset of valvular myofibroblasts may differentiate into an osteoblastic phenotype capable of promoting formation of nodules of calcium and bone. Cartilage in calcified valves indicates that endochondral ossification, similar to that seen in bone fracture healing, is operative. Our results imply that bony metaplasia does not appear to be a late stage in the process of valvular calcification, but rather develops in parallel with it. Supported by the Research Project MZO (Grant 00179906)

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PP3-298 HEPATOMA WITH CARDIAC METASTASIS: AN ADVANCED CANCER REQUIRING ADVANCED TREATMENT Pao-Hsien Chu, Shih-Ming Jung Chang Gung Memorial Hospital, Taiwan Background Hepatocellular carcinoma with cardiac metastasis is very rare and has a poor prognosis. Surgery is frequently reserved for symptomatic patients. This investigation first report the clinical and pathologic findings, and then demonstrates the pathophysiology. Methods This study included eight patients with hepatoma involving the heart, treated by surgical excision. Detailed clinical parameters were reported. Results The patient population comprised two (25%) women and six (75%) men. The mean age of subjects was 50 years old, with an age range of 40 to 70 years old. The presentations included: asymptomatic (75%), heart failure (25%), and pulmonary embolism (12.5%). All lesions involved the right atrium, and extended to the lung (12.5%), inferior vena cava (25%), and left atrium (12.5%). The level of tumor marker, alpha-fetal protein, was not correlated with the severity of metastasis or disease prognosis. Moreover, the availably estimated doubling time was less than 3 months. The pathological findings included variable hemorrhage and necrosis. The survival time following surgery also varied from one month to longer than 30 months. Discussion. Hepatoma metastasis to the heart was detected in all eight patients. This study demonstrates that surgery might help the outcome in such cases. PP3-299 Z DISC PATHOLOGY IN CYPHER-NULL MICE: A 3D ELECTRON MICROSCOPIC STUDY Pao-Hsien Chu1, Anderea Thor2, Ju Chen2, Maryann E. Martone2 1 Chang Gung Memorial Hospital, Thawian 2 University of California, San Diego, USA Comparative studies of the relationship between structure and function offer a promising means of understanding the significance of differences in cytoarchitecture (Not sure what you mean, since you are not studying function?). The threedimensional organization of the sarcomere in the striated muscle of Cypher-null mice, which exhibited dilated cardiomyopathy and skeletal muscle failure, was studied to assess the ultrastructure of the Z disc. Three-dimensional reconstruction of the Z disc was performed on 1 μm thick sections of the diaphragm and heart tissues using electron tomography in both embryonic and new-born Cypher-null mice. Three-dimensional information was extracted from thick sections using stereo pairs and tomographic reconstructions from single axis tilt series from ethanolic phosphotungstic acid (EPTA) staining sections. Oure results show that both the three-dimensional reconstructions and the selectively stained thick sections of the Z disc demonstrated a discontinuous appearance. This investigation used the Telescience Project merging technologies to clearly demonstrate severe fragmentation of the sarcomeric Z disc of embryonic heart and newborn diaphragm of Cypher-null mice, which had not been documented using previous 2D techniques. The analytical results of this investigation strongly confirm that Cypher plays an important role as a linker-strut in the Z disc of striated muscles. PP3-300 MACROPHAGE-SUBENDOTHELIAL EXTRACELLULAR MATRIX CONTACT IN THE ATHEROGENESIS, ELASTIN CONTACT INDUCES TNFĮ UPREGULATION Ichiro Mori1, Tomonori Higuchi2, Misa Nakamura1, Tomoko Wakasa1, Kennichi Kakudo1 1 Department of Pathology, Wakayama Medical University, Japan 2 Department of Pathology, Nara Medical University, Japan

[Background] One of the earliest events of atherogenesis is endothelial injury. When subendothelial tissues are exposed to the blood flow, first exposed is endothelial basement membrane or internal elastic lamina which are mainly composed of elastin, laminin, and collagen type-IV. These extracellular matrices (ECM) are reported to be recognized by elastin-laminin receptor (ELR). We hypothesize that macrophage (Mĭ)-subendothelial ECM contact triggers atherosclerogenesis. We already reported that Mĭs contacted to elastin release M-CSF. This time, we investigated TNF-Į. [Methods] Primary cultured Mĭs were prepared from mouse bone marrow cells. Insoluble elastin and collagen (type I) were suspended in ĮMEM, and applied to adherent Mĭs. Total cellular RNA and protein were extracted, and northern blot analysis and western blot analysis for TNF-Į were performed. To investigate whether this reaction is specific for elastin, we treated Mĭs with galactose, which is said to uncouple the ELR molecular complex, and treated with both elastin and collagen. [Results and conclusion] Mĭs contacted to elastin showed 11.8±1.7 S.D upregulation of TNF-Į mRNA with its peak at 2 hours, and 1.34±0.05 S.D increase of TNF-Į protein with its peak at 6 hours. Type-I collagen also induced TNF-Į mRNA upregulation with its peak at 2 hours. The elastin induced upregulation was inhibited by galactose pretreatment while no inhibition was found in collagen-Mĭ contact. MAP-K inhibition experiment revealed that MEK 1/2 and JNK/SAPK were involved in this reaction while p38 is not. These results strongly suggest that Mĭ-subendothelial ECM contact is a key event of atherosclerogenesis. PP3-301 MORFOLOGICAL STUDY OF CONGENITAL HEART MALFORMATIONS WITH ENDOCARDIAL FIBROELASTOSIS Snezana Duganovska1, Goradana Petrusevska1, Liljana Spasevska1, Miljana Tolovska1, Biljana Dimova1, Roza Kacarska2 1 Institute of Pathology, Medical Faculty, University "St. Cyril and Methodius", Skopje, Macedonia 2 Pediatric Clinic, Clinical Centre, Skopje, Macedonia AIMS: Our study aimed to identify endocardial fibroelastosis (EFE), as a rare but important finding among hereditary heart endocardial malformations, which brings to persistence of fibrous thickening that is a cause of cardiac insufficiency and exitus letalis in early childhood. METHODS: We used autopsy material which has been collected during 19 autopsies, performed at the Institute of Pathology, Faculty of Medicine in Skopje. The patients were neonates and children, not older than 2 years. Histological sections of analyzed hearts were stained with HE, VanGieson-Elastica, Trichrome-Masson and Reticulin-Gomori. RESULTS: EFE was more frequent in males than in females (11/19 cases).Two of the patients showed presence of primary fibroelastosis as an isolated congenital heart malformation; in the rest 17 cases, secondary EFE combined with isolated or complex congenital malformations of the heart and large blood vessels, were found. The analysis of the anatomical location and place of the fibroelastosis in the ventricular system showed that it was more often situated in the left ventricle. More precise morphological analysis has revealed that the endocardium was thickened (to 2mm) along its wall diffusely and had porcelain white color. There was a hypertrophy of the heart, with bigger and wider LV; the apex belongs to the LV, lumen of the LV was wider, interventricular septum was pushed to the RV. The secondary EFE is usually associated with other heart malformations, the endocardium was confluent or focally thickened and accompanied with: SHLS (7 cases); DAP (3 cases); CoAo (2 cases) and pulmonary stenosis, SHDS, ASD, VSD and earlier closing of foramen ovale with one case of each. Histolopatological findings in endocardium showed proliferation of elastic tissue and collagen fibers which were parallel to the

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surface. Inflammatory cells in both endocardium and myocardium were not found. The most frequent cause of death in the cases with EFE was insufficiency of LV (10/19) where severe pulmonary hypertension was found in one case. In the rest of the cases pulmonary hemorrhage 2/19 was present, bronchopneumonia in 2/19, aspiration in 1/19, enterocolitis in 1/19, brain hemorrhage in 1/19 and sepsis in 1/19 cases. CONCLUSIONS: Morphological analysis as well as histopathological findings has shown that most of the analyzed complications were due to mechanical obstruction of the blood flow. The most frequent was secondary EFE accompanied with other CHM, which was the cause of: "inlet" and outlet" obstructions; anoxia in utero; earlier closing of the FO. PP3-302 INDUCTION OF MITOCHONDRIAL BIOGENESIS IS A MALADAPTIVE MECHANISM IN CARDIAC REMODELING Mariangela Sebastiani1, Carla Giordano1, Claudia Travaglini1, Massimo Zani1, Chiara Nediani2, Elisabetta Borchi2, Massimiliano Mancini3, Robert Taylor4, Pietro Gallo3, Giulia D'Amati3 1 Dipartimento di Medicina Sperimentale, Sapienza, Universita’ di Roma, Rome, Italy 2 Dipartimento di Scienze Biochimiche, Università di Firenze, Florence, Italy 3 Dipartimento di Medicina Sperimentale, Sapienza, Universita’ di Roma, Rome, Italy. 4 Mitochondrial Research Group and Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, United Kingdom Objectives. The purpose of this study was to clarify the molecular mechanisms linking mtDNA dysfunction to cardiac remodeling. Background. Defects of the mitochondrial genome cause a heterogeneous group of clinical disorders, including mitochondrial cardiomyopathies (MIC). The molecular events linking mtDNA defects to cardiac remodeling are unknown. Both energy derangements and increase of mitochondrial-derived reactive oxygen species (ROS) could play a role in the development of cardiac dysfunction in MIC. In addition, mitochondrial proliferation could interfere with sarcomere alignment and contraction. Methods. We performed a detailed morphologic and molecular analysis on failing hearts from three patients with MIC, failing human hearts due to ischemic heart disease (IHD) or dilated cardiomyopathies (DCM), and nonfailing hearts. Results. MIC hearts showed marked mitochondrial proliferation, with myofibril displacement. Consistent with morphologic features, increase in mtDNA content per cell and induction of genes involved in mitochondrial biogenesis, fatty acid metabolism and glucose transport were observed. Downregulation of these genes characterized DCM and IHD hearts. Furthermore, up-regulation of uncoupling proteins and a pronounced increase in mitochondrial-derived ROS was observed in MIC as compared to failing hearts due to other etiologies. The observed increase in free radical production was not paralleled by the induction of antioxidant enzyme activity, despite marked upregulation of the corresponding genes. Conclusions. Our results suggest that mitochondrial biogenesis is a maladaptive response in MIC and, possibly, in other metabolic cardiomyopathies. In fact, increased oxidative stress, mechanical interference with sarcomere alignment and contraction, and, possibly, uncoupled respiration are detrimental factors to myocytes function, besides energy deficiency.

PP3-303 COULD SUDDEN CARDIAC DEATHS HAVE BEEN PREVENTED: AN AUTOPSY STUDY Patrick Gallagher1, Hayley Burnley1, Debbie Chase2, Paul Roderick2, Paul Poberts3, John Morgan3 1 Pathology, Southampton University Hospitals, Southampton, United Kingdom 2 Public Health Medicine, Southampton University Hospitals, Southampton, United Kingdom 3 Wessex Cardiac Unit, Southampton University Hospitals, Southampton, United Kingdom The objective of this study was to identify which, if any, sudden cardiac death (SCD)victims could have been identified before their death and considered for an implantable cardioverter defibrillator (ICD) on the basis of their previous cardiac history.Autopsy proven cases of SCD were identified over a one year period in a defined population. They were categorised by a modification of Davies method. Those with evidence of myocardial scarring, coronary narrowing or ventricular hypertrophy may have died as a result of a ventricular arrhythmia. However cases with clear evidence of acute coronary thrombosis or acute infarction were excluded. Hospital and GP records were examinaed for information on previous symptoms, investigations and past history of cardiac disease. Two cardiac electrophysiologists judged the appropriateness of each case against pre defined criteria. 366 of 975 post mortems were sudden cardiac deaths and in 232of these there was no evidence of acute thrombosis or infarction. 215 lived within the catchment area, 54% were male and the median age was 75 years. Agreement between experts was good (kappa score 0.64). One case was considered appropriate for ICD insertion. 41% of cases had no evidence of cardiac events or prior heart disease. 49% of the remainder had previous cardiac events or symptoms suggestive of cardiac arrhythmias but had not been referred for further investigations. In particular some patients with a previous myocardial infarction were not referred for 24 hour ECG tracings and some of those with suspected heart failure did not have echocardiography. Our study has shown that 40% of SCDs could not have been predicted A significant proportion of the remaining patients were not referred for appropriate investigations. PP3-304 APOPTOSIS-RELATED FACTORS P53, BCL-2 IN ACUTE AND CHRONIC ISCHEMIC CARDIAC DISORDERS Angelos Tsipis, Anna Maria Athanassiadou, Pauline Athanassiadou, Nicolaos Kavantzas, George Agrogiannis, Efstratios Patsouris Department of Pathology, Medical School, University of Athens, Greece Background: Cardiac myocyte death during ischemic injury has been thought to occur exclusively by necrosis, but recently several studies have demonstrated that large numbers of myocytes undergo apoptosis in response to ischemic disorders. P53 limits cellular proliferation by inducing cell cycle arrest and apoptosis in response to cellular stresses such as DNA damage, hypoxia, and oncogene activation. P53 mediates apoptosis through a linear pathway involving bax activation, cytochrome c release from mitochondria, and caspase activation. The Bcl-2 family of proteins constitutes a critical checkpoint in cell death. These proteins contain agonists and antagonists of apoptosis, and alterations in their ratio determine the life or death of a cell. Proapoptotic proteins include Bax, Bak, Bad, and Bcl-xs whereas Bcl-2 and Bcl-xL are antiapoptotic. The aim of this study was to investigate the expression of apoptosis-related proteins p53, bcl-2 in acute and chronic ischemic cardiac disorders and their association with myocyte death in ischemic hearts. Method: We studied myocardial samples of hearts with histologic findings of acute myocardial infarction (group A, n=15), old myocardial

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infraction (group B, n=15), and chronic ischemic heart disease (group C, n=15). Myocardial samples of normal heart were also included in this study (control group, n=15). An immunohistochemical method was performed with the use of p53 and bcl-2 antibodies, in order to investigate the expression of apoptosis-related proteins p53, bcl-2 in ischemic cardiac disorders. Results: The percentage of bcl-2 positive samples was 66% in acute myocardial infarction (Group A, 10/15 positive samples). In old myocardial infraction and chronic ischemic disease the bcl-2 positive samples demonstrated weak staining as in the control group. P53 positive samples was 20% in acute myocardial infraction (Group A, 3/15 positive samples), 80% in old myocardial infraction (Group B, 12/15 positive samples), 80% in chronic ischemic heart disease (group C, 12/15 positive samples) and 20% in control group (3/15 positive samples). Conclusions: The elevated expression of p53 is associated with the progressive loss of myocytes by apoptosis and may play a role in the evolution of the chronic heart failure in patients with old myocardial infraction and chronic ischemic disease. The increased expression of bcl-2 in acute myocardial infraction represents a possible compensatory mechanism of salvaged myocytes. PP3-305 THE VASORELAXATION OF RAT RENAL ARTERY INDUCED BY WINE POLYPHENOL RESVERATROL Ljiljana Gojkoviü-Bukarica1, Jasmina Markoviü-Lipkovski2, Helmut Heinle3 1 Institute of Clinical Pharmacology, Pharmacology and Toxicology, Faculty of Medicine, Serbia 2 Institute of Pathology, Faculty of Medicine, Belgrade, Serbia 3 Institute of Physiology, Tubingen, Germany BACKGROUND: Resveratrol, a stilbene polyphenol found in grapes and red wine, has recently been found to produce vasorelaxation in endothelium-dependent and endotheliumindependent manner. The aim of this study is to determine the mechanism(s) of relaxation produced by resveratrol in the isolated rat renal artery (RA). METHOD: RA rings were precontracted with phenylephrine. In order to assess the endothelial integrity of the preparation we have used acetylcholine. Endothelium was removed mechanically by rubbing with a steel wire. Failure of arteries to relax to acetylcholine was considered as indicator of state of endothelial denudation. For detection of K+ channels in smooth muscle, peptide-specific antibodies in immunoperoxidase were used. RESULTS: Resveratrol (1–100 microM) produced concentrationdependent relaxation of RA rings with endothelium (EC50~10 microM, n=10) and without endothelium (EC50~15 microM, n=12). Methylene blue and L-NAME did not antagonize the resveratrol-induced relaxation of RA rings with endothelium. In order to analyze the contribution of different types of K+ channels in resveratrol-induced relaxation in the RA, various K+ channel blockers were used. The relaxation of RA was not blocked by glibenclamide, a selective ATP-sensitive K+ channel blocker, and tetraethylammonium, a non-selective blocker of calcium-dependent K+ channels. 4-aminopyridine, non-selective blockers of voltage-dependent K+ (Kv) channels, antagonized resveratrol-induced relaxation of RA in a noncompetitive manner. Margatoxin, highly selective blockers of Kv1.1-1.6 channels shifted the concentration response curves induced by resveratrol to the right without significant inhibition of maximal responses. Kv1.3 channels were detected in endothelium but not in the smooth muscle of RA using peptide-specific antibodies in immunoperoxidase. CONCLUSION: It is likely that endothelial Kv1.3 channels are involved in relaxation of RA produced by resveratrol. However, further experiments with smooth muscle peptide-specific antibodies need to be performed.

PP3-306 FATAL DRUG INDUCED HYPERSENSITIVITY MYOCARDITIS AFTER MINOCYCLINE-THERAPY Peter Dorfmuller1, Olivier Chosidow2, Francois Cook2, Jean-Louis Trouillet3, Pascal Leprince4, Eva Comperat1, Frédérique Capron1, Isabelle Brocheriou1 1 Service d’Anatomie et de Cytologie Pathologiques, Groupe Hospitalier Pitié-Salpêtrière, Université Pierre et Marie Curie, Paris, France 2 Service de Dermatologie, Hôpital Tenon, Université Pierre et Marie Curie, Paris, France 3 Service de Réanimation Médicale, Groupe Hospitalier PitiéSalpêtrière, Université Pierre et Marie Curie, Paris, France 4 Service de Chirurgie Thoracique et Cardio-Vasculaire, Groupe Hospitalier Pitié-Salpêtrière, Université Pierre et Marie Curie, Paris, France Background: Acute Hypersensitivity Syndrome (HSS) is an allergic-type complication of drug therapy with associated symptoms such as rash and fever associated with bloodeosinophilia and multiorgan involvement. Allopurinol, anticonvulsants, sulphonamides and tetracyclines are amongst the most frequent causative agents. A genetic predisposition has been associated with HSS which shows a higher overall-prevalence in the African-American population, especially with minocycline. Visceral lesions such as interstitial pneumonitis, hepatitis, interstitial nephritis or myocarditis can be lethal. Case and clinical findings: A 34-year-old black African woman, without any prior medical history, was admitted to the emergency unit in October 2006 with fever, arthralgias and dyspnea. She had been treated by minocycline 2 weeks before admission for chlamydial genital infection. The physical examination showed tachycardia, erythematous papular exanthema on arms, erythematous angina, facial oedema and leucocytosis. No infectious source was identified. Drug induced HSS after minocycline was suspected and she received high dosed corticotherapy. After initial improvement of her dyspnea, she was discharged after 1 week, against medical advice, and stopped her medication. One month later, she was readmitted with severe hypotension, dyspnea and major left ventricular systolic dysfunction. Support by a central extracorporeal-membrane-oxygenation was provided during open-heart surgery and myocardial biopsy was performed. She died 6 days later after multiple organ failure. Microscopic findings: Myocardial biopsy showed mixed inflammatory infiltrates with interstitial and perivascular distribution, mainly consisting in lymphocytes, plasmocytes, histiocytes and an increased proportion of eosinophiles. This acute eosinophil-rich myocarditic pattern was associated with pronounced interstitial edema and recent necrosis of myocytic fibers explaining myocardial pump failure. Conclusion: Eosinophilic myocarditis (EM) may occur in the setting of a hypereosinophilic syndrome, parasitic infections, malignancy or vasculitis. EM has been sporadically described in HSS, but the majority of reports are post-mortem observations due to frequent fatal outcome. Minocycline is often prescribed as the first-line therapy in acne or genital infection. Although fatal side effects are rare, HSS should be taken into consideration by the pathologist when confronted to eosinophil-rich necrotizing myocarditis. In our case, endomyocardium biopsy has been helpful for the diagnosis which often remains unsuspected. PP3-307 KAWASAKI DISEASE Zvezdana Dolenc - Strazar, Klara Limbaeck - Stokin Institute of Pathology, Medical Faculty, University of Ljubljana, Slovenia Introduction: Kawasaki disease (KD) is acute self-limited vasculitis effecting mainly children under 5 years. Etiology is still unknown. Currently debate is about infectious trigger leading to

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self-directed immune response. Coronary arteries are predominantly involved; aneurysms may lead to sudden death, infarction and ischemic heart disease. Treatment in the acute phase is directed towards reducing inflammation of the artery wall and preventing thrombosis, at the late phase against myocardial ischemia and infarction. Case report: Three and a half year old boy was admitted to hospital, four days after he was taken ill, with fever and classical symptoms of KD. Viral tests were negative. Among usual therapeutics, he was treated with aspirin and repeated doses of iv-immunoglobulin, without success. Echocardiography showed diffuse dilatation of coronary arteries, which persisted till the death. After three and a half months of hospitalizations, after receiving infliximab, fever dropped and laboratory tests normalized for the first time, but he died 10 days later, four months after onset of the illness, with signs of myocardial infarction. At autopsy the heart was enlarged. Histology showed acute myocardial infarction of left ventricle with some older necrotic areas with granulation tissue. Main coronary arteries were diffusely dilated, diameter of RCA aneurysm was 8 mm, and of others between 3-4 mm. Due to progressive myointimal proliferation aneurysms had very reduced lumen diameter. In distal part of coronary arteries there were organized thrombi in recanalisation. Beside bronchopneumonia, hemorrhagic pulmonary infarction and cyanosis of internal organs, there were no other major abnormalities. Conclusion: Clinical picture of typical KD is characterized by fever and at least four of five principal features which are rash, conjuctival injection, cervical lymphadenitis, inflammation of the lips and oral cavity, swelling of the hands and feet. Especially in infants, some of the criteria are lacking. It is very important to diagnose these cases early, as they are at risk of developing coronary artery aneurisms and stenosis. Myocardial infarction is the principal cause of death and may occur in acute phase, but commonly year or even years later, especially in patients with giant aneurysms when measuring more than 8 mm in diameter. Surgical bypass procedures are effective when stenoses are proximal, what was not the case in our patient. There were reports in literature that a small group of KD patients can benefit from transplantation, with no report of recurrence. PP3-308 CLINICAL AND PATHOLOGICAL FINDINGS IN FASTPROGRESSING OBSTRUCTIVE HYPERTROPHIC CARDIOMYOPATHY (OHCM) IN CHILDREN AND ADULTS Alexandra Gudkova1, Margarita Rybakova1, Konstantin Borisov2, Evgeny Shlyakhto1, Leo Bokeria2 1 Pavlov State Medical University, Department of Faculty Therapy, Saint-Petersburg, Russia 2 Bakulev Research Center of Cardiosurgery, Moscow, Russia Aim: To compare clinical and pathological features in severe cases of obstructive hypertrophic cardiomyopathy in children and adults. Materials and methods. Nineteen children (of them, 17 survived and 2 died) and forty-seven adults (30 survived and 17 died) with OHCM were observed. OHCM diagnosis was verified by means of echocardiography, MRI, macro- and micromorphometry of affected tissues. Myocardial samples were taken during cardiac surgery or urgent autopsies. Microscopic preparations were stained with H&E, or van Giezon stain. The degree of disturbed tissue architecture, as chaotic rearrangement of cardiomyocytes, was estimated by a semiquantitative method using a 5-point morphometric score (M. Sutton et al., 1980). Results. Mid-ventricular OHCM location in pediatric group was as common as 63,2 % (12/19). In adult patients, this variant was diagnosed in 40.4% of cases (19/47). In general OHCM group, mid-ventricular OHCM was registered in 31 of 66 patients (46,7 %). Transmural myocardial scars have been revealed in 4 of 10 (40 %) autopsied HCM patients with mid-ventricular OHCM, and in 2 of 7 (28.5 %) deceased HCM patients with left ventricular

outflow obstruction. In three adult patients with mid-ventricular OHCM, an apical aneurysm of top area was revealed, in absence of significant hemodynamic coronary stenosis, as evidenced by coronarograpy. In pediatric OHCM, a degree of cytoarchitectonic disorders in the right half of interventricular septum varied from 2.25 to 3.00 (2.87±0.05) units, thus being higher than at adults (p <0,001). In adult patients, this parameter varied from 0.25 to 3.00 (1.92±0.10). Stromal portion of the right part of interventricular septum in the children varied from 1.08 to 6.67 % (3.62±0.30). In adults with OHCM, this parameter changed from 2.12 up to 10.60 % (5.36±0.34), thus being higher than in pediatric patients (p <0.005). Conclusions. Increased grade of tissue «disarray» is a sufficient feature of OHCM in children. Meanwhile, enlarged area of myocardial stroma is a significant sign of OHCM progression in adults. In the patients with severe variants of the disease, occurrence of mid-ventricular OHCM is comparable to incidence of HCM with left ventricular outflow tract obstruction. Mid-ventricular OHCM progression is more often complicated by transmural myocardial scars. Higher frequency of apical aneurysm, developing at the top area, is detectable in adult patients with mid-ventricular OHCM. PP3-309 RELATION OF MORPHOLOGICAL STRUCTURE OF HIBERNATING MYOCARDIUM TO CONTRACTILE RESERVE IN HUMANS Elena Stefanovic1, Petar Otasevic1, Ljiljana Lausevic-Vuk1, Predrag Milojevic1, Bosko Djukanovic1, Milena Jovic2, Lidija Zolotarevski2 1 Dedinje Cardiovascular Institute, Belgrade, Serbia 2 Military Medical Academy, Belgrade, Serbia Objective.The aim of this study was to investigate the morphologic characteristics of the hibernating human myocardium and to correlate its with dobutamine stress echocardiography (DSE). Methods and results.We evaluated 15 patients with coronary disease (58 +/- 12 years old, ejection fraction 38+/-14 %) with a corresponding wall motion abnormality on DSE (up to 10 micrograms kg (-1) min(-1) before coronary bypass surgery. During surgery, transmural myocardial biopsies from hypokinetic or akinetic area were perfomed (n=15). The samples of myocardium were analyzed by histopathology and immunohistochemistry to investigate the extent of interstitial fibrosis, intracellular and interstitial proteins. Among the 15 biopsied segments included in the study, 7 recovered function as assessed with DE (an echocardiography) one month after bypass surgery. Segments with DE viability showed less fibrosis and less vimentin expression, more glycogen, a higher ration of alphasmooth muscle actin, actin and desmin then those without recovery. The degree of severity of the morphological changes (three stages) correlated well with the demonstration of inotropic reserve during DSE and with the extent of postoperative functional recovery (wall-motion score index, NYHA ). Conclusion. Morphologic evidence of hibernating myocardium correlates with DSE findings wich has high diagnostic accuracy for the detection of myocardial viability. PP3-310 THE ASSOCIATION FOR EUROPEAN CARDIOVASCULAR PATHOLOGY Ulrik Baandrup1, Cristina Basso2, Patrick J Gallagher3, Gaetano Thiene2, Allard Van Der Wal4 1 Aarhus, Denmark 2 Padua, Italy 3 Southampton, United Kingdom 4 Amsterdam, The Netherlands Although there are comparatively few pathologists who specialise in cardiovascular pathology all practicing pathologists are aware of the great importance of cardiovascular diseases. Ischaemic

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heart disease, stroke and peripheral vascular disease are major causes of morbidity and mortality in all societies. Much of the improvement in life expectancy in the western world has been the direct result of improved prevention and treatment of these disorders. The Association for European Cardiovascular Pathology is an affiliated society of the European Society of Pathology. As a professional association it is committed to setting the standards of practice throughout Europe and providing post graduate education and training in cardiovascular pathology. We alternate our annual meetings between the European Society of Pathology and free standing meetings held in the late autumn. Our last meeting held in Aarhus, DK in November 2006 included three separate symposia, invited keynote presentations and clinicopathological presentations. In Istanbul we are organising a Surgical Pathology slide seminar on Cardiovascular Disease and a Symposium on Sudden Cardiac Death. We extend a warm invitation to all delegates to attend our evening meeting On Monday 10th September where a series of clinicopathological cases will be presented. The poster will describe the activities of our Association, the format and venue of future meetings and educational opportunities in Cardiovascular Pathology PP3-311 REACTIONS OF MYOCARDIUM TO THE PRESENCE OF IMPLANTABLE ELECTRONIC MEDICAL DEVICES Katarina Múþková1 , Sepši Milan2, Šindler Martin3, Souþek OndĜej1, Kir Michal4 1 Departement of Pathology, Faculty Hospital Brno, Czech Republic 2 Departement of Internal Medicine and Cardiology, Faculty Hospital Brno, Czech Republic 3 Departement of Forensic Medicine, St. Anne's Faculty Hospital Brno, Czech Republic 4 Institute of Biostatistics and Analyses, Masaryk University Brno, Czech Republic Background: In view of constantly increasing number of electronic medical devices (EMDs) implantations, the possible alterations of cardiac structures associated with the presence of EMD electrodes are studied. Although transvenous implantable leads are coated with inert materials to minimize the reaction of human tissues, the experiment on dog hearts showed inflammatory and degenerative changes developing in the atrial wall after implantation of pacemaker (Pacing Clin Electrophysiol. 2003 Mar;26(3):685-91). Methods: The hearts of 182 autopsy cases (130 males, 52 females ) with EMDs (pacemakers and implantable cardioverter-defibrillators) were examined. The mean patients´ age at the time of EMD implatation was 76,8(±9,8). Different anatomic parameters of each heart were registered together with the number of fixations of leads to various cardiac structures. The tissue samples obtained from places of electrode insertions and each heart ventricles were processed using standard histological techniques. The tissue sections stained with hematoxylin-eosin were examined microscopically to identify inflammatory reaction, the presence of granulation tissue, fibrosis, hyaline scar, calcification and other changes in myocardium adjacent to the lead. Results: The mean weight of examined hearts was 544,48 g (±155,80), the mean left ventricle thickness was 16,22 mm (±4,19), the mean right ventricle thickness 4,86 mm (±1,94). 70% of all leads were attached to the cardiovascular structures by the band of fibrous tissue. 49,1% of all leads were fixed to the tricuspid valve or some of papillary muscles (potentially harmful fixation). The histological examination showed fibrosis and hyaline scar tissue in the electrode-myocardial interface. There was no correlation between the point of the fixation of lead and macroscopic parameters of right and left ventricles. The type of leads (pacing versus defibrillator), the type of stimulation (unipolar versus bipolar), and the percentage of stimulation did not influence the character of reaction in places of insertions. The mild reduction

of the tricuspid valve circumference was observed in cases with long-time insertion of the lead. The occurrence of thrombi in heart cavities was minimal (2%)in association with electrodes. Conclusion: Implantation of EMD is followed by a reaction of the myocardium adjacent to the place of insertion. Potential harmful effect of EMDs was observed in cases with a long- time insertion of the lead. The research is supported by grant MH CZ 1A/8598-3. PP3-312 CARDIAC MYXOMA WITH GLANDULAR COMPONENT: CASE REPORT Arzu Tasdemir 1, Ozlem Canoz 1, Kutay Tasdemir 2 1 Erciyes University Medical Faculty Department of Pathology, Turkey 2 Erciyes University Medical Faculty Department of Cardiovascular Surgery, Kayseri, Turkey Cardiac myxoma is the most common primary cardiac tumor. Glandular epithelial differentiation is a rare phenomenon. A striking observation, seen in approximately 1-5% of cardiac myxomas. Here, we present a case of a cardiac myxoma with glandular component removed from the left atrial wall of a 38year-old woman. Cardiac myxomas are the most common primary cardiac tumor in adults. Myxoma constitute approximately 50% of primary tumors of the heart. The sporadic tumor occurs in middle-aged woman, usually in the left atrium. A glandular component in cardiac myxoma is uncommon. The histogenesis of this feature is enigmatic. The presence of the epithelial element as glandular structure is an example of one spectrum of the multidirectional differantiation. Alternative cardiac myxoma was described as entrapped foregut rest or intracardiac endodermal heterotopia in some literatures. The results suggest that cardiac myxomas is the occurence of a glandular component embedded within the myxomatous stroma. Malignancy of the glandular component in cardiac myxoma has been reported, but in many cases this needs to be set apart from tumor embolization and local recurrence. PP3-313 SYPHILITIC AORTITIS WITH RUPTURED THORACIC AORTIC ANEURYSM, A CASE REPORT Khalid Al-Shibli, Hedda Soloy-Nilsen Pathology Department, Nordland Central Hospital-Norway Background: Aortic aneurysm is an important cause of morbidity and mortality. The symptoms caused by it can mimic other diseases and hence it is usually under diagnosed. Therefore, ruptured thoracic aortic aneurysm is a not uncommon postmortem finding. Syphilitic aortitis is a rare entity nowadays, but cases are still seen and its diagnosis needs a high index of suspicion. We report her post-mortem case of ruptured thoracic aortic aneurysm due to tertiary syphilis. Methods: We report post-mortem findings of a 71 years old man, who developed severe chest pain with vomiting and collapsed at home. man developed severe chest pain with vomiting and collapsed at home. Resuscitation was done with no benefit and the patient died after 30 minutes from reaching the hospital. No history of diabetes or systemic hypertension. Results: Post-mortem examination revealed a ruptured thoracic aortic aneurysm measuring 12x12cm involving the arch and descending parts of the aorta with hemothorax. The aortic valve showed mild dilatation and the same for the left ventricle. The heart weight was 475gm. No significant atherosclerosis was noted in any part of the aorta or in other major arteries. Microscopic examination of the aorta showed chronic inflammatory infiltrate, most sever in the adventitia and media with large number of plasma cells mostly around the vasa vasorum with endarteritis obliterans picture. Microscopy showed no significant atherosclerosis or cystic medial degeneration. The abdominal part of the aorta was

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not dilated and showed no significant atherosclerotic changes. Post-mortem examination revealed no features of Marfans syndrome or connective tissue diseases. Conclusion: Syphilis continues to be a cause of significant morbidity and mortality. It is a great mimicker for many diseases in its different stages. Missing a diagnosis will deprive the patient from a chance of an effective and specific treatment with a high chance of cure. High index of suspicion is required for the diagnosis of aortic aneurysm as a cause of chest and abdominal pain; and tertiary syphilis should be considered as a possible cause for thoracic aortic aneurysms. PP3-314 CAUSE, FREQUENCY AND TIME OF ONSET OF ASYMPTOMATIC ACUTE MYOCARDIAL INFARCTION IN ABRUPT UNEXPECTED DEATHS IN A FORENSIC AUTOPSY MATERIAL Elin Mortensen1, Torleiv Rognum2, Bjørn Straume3, Leif Jørgensen4 1 Dept fo Pathology, University Hospital of North Norway, Tromsø, Norway 2 Insitute of Forensic Medicine, University of Oslo, Norway 3 Institute of Community Medicine, University of Tromsø, Norway 4 Department of Pathology, University of Tromsø, Norway Background: Acute myocardial infarction is frequently found at autopsy in persons dying suddenly. The aims of this study was to determine the time between onset of an asymptomatic myocardial infarction to the time of abrupt deaths and to determine whether there are morphological signs of spasm of the epicardial coronary arteries at autopsy in persons dying abruptly and unexpectedly. Methods: In a forensic autopsy service cases of sudden deaths were selected consecutively, one group with a preliminary diagnosis of coronary heart disease, and a control group with cases of a non-coronary cause, mainly self-inflicted death. Complete autopsy was carried out with particular emphasis on the heart examination. Without knowledge of which group the case belonged, multiple histological sections from the coronary arteries and the myocardium were examined. The age of the lesions was estimated by observing several time-related phenomena: macroscopic evidence of myocardial infarct or, microscopic presence of C9-positivity, and increasing quantity of CD15 positive leucocytes in sections from the myocardium. The degree of spasm is quantified by picture analysis, measuring the degree of folding of the internal elastic lamina in cross sections of elastin stained proximal and distal part of the coronary arteries. Results: Acute coronary lesions (rupture of necrotic plaque, thrombi, etc.) were found in 13 coronary cases, and myocardial infarcts, with onset within 5-6 hours prior to death in 13 cases, no infarct in 2 cases. In the non-coronary group, acute coronary changes were found in 4 cases, small recent infarcts in 7. The degree of folding was significantly greater in the distal section of right coronary artery in cases of the coronary group compared to the folding in the same section of the non-coronary cases. There were no significant differences in the other arteries. Conclusion: Sudden, unexpected deaths in persons with coronary disease occur within the first several hours after the onset of an asymptomatic myocardial infarction. Small, acute infarcts were found incidentally also a minority of the non-coronary group. Also,our findings indicate that a spasmic contracture of an artery may be diagnosed post mortem. In several of the coronary cases spasm of the distal part of the right coronary artery may have caused focal ischemia in the central parts of the cardiac conducting system precipitating a lethal arrhythmia.

PP3-315 AN EXPERIMENTAL MODEL OF MYOCARDIAL INFARCTION IN PIGS: REPERFUSION DAMAGE, DYNAMICS OF HEALING PROCESSES Leos Kren1, Zdenek Pavlovsky1, Jiri Mayer1, Jaroslav Meluzin1, Ladislav Groch1, Petr Rauser2, Michal Vlasin2, Ivan Hornacek1 1 University Hospital Brno, Czech Republic 2 University of Veterinary Medicine Brno, Czech Republic Background: The goal of experiments was to create an animal model of myocardial infarction and to evaluate ischemic/reperfusion damage and dynamics of healing processes. The prospective use of the model is to evaluate fate, quantity and the best way of application of stem cells in the management of myocardial infarction. Material and Methods: Four experimental animals, female pigs, with weight from 30 to 40 kg were used. An experimental myocardial infarction was performed via occlusion of r. interventricularis anterior for 40 minutes. The hearts were examined 1 hour, 3 days, 5 days and 7 days after the procedure. Results: Macroscopically, red infarction, characteristic for ischemia with reperfusion was found. Microscopically, healing process with granulation tissue production/collagen deposition was remarkably accelerated in coparison with data in the literature. Also, besides of deposition of intracellular calcium, large deposits of extracellular calcium were found. Conclusion: dynamics of healing processes in myocardial infarctions in young female pigs is remarcably accelerated compared to data in the literature (probably because of examining tissues from significantly older human patients described in the literature). Also, not well described is a fenomena of large deposition of extracellular calcium, which may possibly interfere with healing processes. PP3-316 PAPILLARY FIBROELASTOMA WHICH OCCURED IN THE POSTERIOR LEAFLET OF THE MITRAL VALVE: REPORT OF A CASE Mihriban Gurbuzel, Saime Gul Barut Haseki Education and Research Hospital, Istanbul, Turkey Papillary fibroelastoma (PFE) is a rare benign tumor arising from the cardiac endotelium.They are usually located on valves, mostly the aortic valve.These tumors can be responsible for strokes, transient ischemic attacs, anjina pectoris, infarction and sudden death.Echocardiographic diagnosis, followed by surgical excision, may prevent these complications.Here, we report a rare case of papillary fibroelastoma because of its uncommon apperance in the posterior leaflet of the mitral valve. CASE: A 33-year-old male was hospitalized with a diagnosis serebrovasculer accident.Echocardiography revelaled a mass attached to the posterior mitral leaflet.The mass was removed by surgery.It was 0.7x0,6x0,5cm. diameter graywhite color and contain small papillary projections. On the microscopic evaluation,there was a lining of endocardial cells covering a core of hyalinized hypocelluler stroma. We identified PFE and supported immunhistochemically. The endotelial cells on papillary projection are reactive for CD34 and the stromal cells are weakly reaktive for S100. DISCUSSION: Papillary fibroelastomas are primary cardiac tumors of valvuler tissue. They are usually seen in elderly patients. But PFE of the mitral valve in 3-year-old child has also been reported. Transthorasic echocardiography may lead to suspicion of a PFE, but transoesophageal echocardiograhy is required for confirmation. It is identified histologically. This tumor may occur as isolated lesion or concomitant valve stenosis or other cardiac abnormalities. It is an important source of emboli. The lesions consist of a slender or broad fibrocollagenous stalk from which numerous papillary villous projections. Promt surgical excision is indicated in most cases. Anticoagulation is only recommended in situations of high surgical risk and during the wait for surgery.

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PP3-317 MORPHOLOGICAL CHANGES OF CEREBRAL VASCULAR STRUCTURES IN HYPERTENSIVE PATIENS DEAD WITH INTRACEREBRAL HAEMORRHAGE Iancu Emil Plesea1, Stelian Danut Enache2, Corneliu Cristian Georgescu1, Mirela Corina Ghilusi2, Dan Cioroianu1, Oltin Tiberiu Pop1, Alexandru Camenita2, Mihaela Tenovici3, Cornelia Enache2 1 University of Medicine and Pharmacy Craiova, Romania 2 Emergency County Hospital Craiova, Romania 3 National Railways System Universitary Hospital Craiova, Romania Background The purpose of this study was to assess the spectrum of cerebral vascular wall (VW) changes in patients with clinical suspicion of primary intraparenchymal hematoma, confirmed after autopsy. Method 82 cases, clinically diagnosed with hypertension (HT), died with stroke and confirmed with intracerebral haemorrhage at autopsy were selected. The studied material consisted of nervous tissue situated near and distant from the haemorrhagic focus (HF). The specimens were processed following the classical histological technique and stained with usual stainings (H-E, van Gieson and Goldner trichromes) and immunohistochemical stains for basement membranes (BM) (Collagen-IV) and endothelial cells (CD34). Results Extraparenchimal large arteries showed only classical atheroslerotic lesions (AthL) which were found in most of the patients. The severity and extention of AthL presented wide individual and interindividual variations and were not correlated with the patients’ age. The atrophy of the middle layer resulted in significant loosening and subsequent aneurismal spindle-shaped dilatations in certain extracerebral arterial segments. In some cases, thrombi in the lumen of large arteries were found, which partially occluded them. Intraparenchymal arteries and arterioles showed the entire range of VW degeneration steps caused by HT: thickening and folding of the inner elastica, hypertrophy followed by degeneration of the middle smooth muscle layer (SML), substitutive progressive and extensive fibrosis, starting from the outer adventicial layer and hyaline degeneration (HyD) of the collagen fibres which replaced the middle and outer VW layers. All observed changes had a focal irregular distribution, not related with the proximity of HF. Capillary walls showed focal or circumferential thickening of BM caused by the encreasing of collagen-IV material, followed by HyD of collagen fibres. The CD34 immunostaining showed that endothelial cells kept their structural integrity. Spindle-shaped dilatations and rupture of impaired VW in areas with hyalinization were also observed. Conclusions The VW changes are different depending on vessel histological structure and function. The sequence of cerebral VW degenerative lesions results in HyD of excessive fibrillar material from arteriolar wall or from BM. Weakening of wall resistance by hyalin material and minimal resistance of the surrounding parenchyma, explains vascular rupture due to HT, with subsequent HF only in cerebral parenchyma. The more adequate term for describing the VW changes seems to be sclerosis with HyD. PP3-318 ARRHYTHMOGENIC RIGHT VENTRICULAR CARDIOMYOPATHY AS A CAUSE OF SUDDEN DEATH IN 12 YEARS OLD GIRL Jordanka Brachkova Medical University Pleven, Bulgaria BACKGROUND: Arrythmogenic Right Ventricular Cardiomyopathy (ARVC) has recently been recognized as a cause of ventricular tachycardia and sudden death in young people. According to the WHO, ARVC is distinct from the more familiar forms of cardiomyopathy (dilated, hypertrophic and

restrictive) and is characterized by three features: progressive fibrofatty replacement of right ventricular myocardium, strong familial transmission and presentation with symptomatic arrhythmias or sudden death. Although a gene defect was localized on chromosome 14, the pathogenesis remains obscure. The disease almost always occurs after puberty. METHODS: Autopsy and routine histology. CASE: Twelve years old girl was operated for ductus thyreoglossus persistens. Two hours after operation she started bleeding from operational wound. Despite of chemostasis and reanimation procedures , she died three hours later. Autopsy investigation revealed : Thymic follicular hyperplasia (95gr.) and adrenals atrophy ; Degenerative changes with apoptotic body between cardiomyocytes and its replacement by fibrous scars; Subendocardial and interstitial fibrosis and edema in right atrium wall and large amounts of fat in right subepicardium and around coronary vessels was seen as well. The right ventricular wall was severely thinned with extensive fatty infiltration and loss of myocytes. Subepicardial fatty tissue and myocardial fatty tissue replacement was most severe in apical parts of right ventricle. Thymic follicular hyperplasia was accompanied with necrotic Hassal’s body and many eosinophyles and neutrophyles in medulary sone. Edema and inflammatory cells infiltrates composed from eosinophils, plasma cells, lymphocytes, neutrophils and mast cells was in subepithelial connective tissues of pharynx and epiglottis CONCLUSION: Cause of death was shock which first clinical symptom was bleeding in site of operation. Bleeding is consequence of acute congestive heart failure due to arrithmogenic right ventricular cardiomyopathy. In this case ARVC is accompanied with thymic hyperplasia, adrenal glands atrophy and ductus thyreoglossus persistens, which are probably involved in pathogenesis of ARVC. PP3-319 EXPRESSION OF TELOMERASE AND Ki-67 IN MYOCYTES AFTER MYOCARDIAL INFARCTION Angelos Tsipis, Anna-Maria Athanassiadou, Pauline Athanassiadou, Nicolaos Kavantzas, George Agrogiannis, Efstratios Patsouris Department of Pathology, Medical School, University of Athens, Athens, Greece Background: The presence of myocyte proliferation in the adult heart remains controversial. Telomerase replaces telomeric repeat DNA lost during the cell cycle, restoring telomere length. This enzyme is present only during cell replication and its activity reflects the extent of proliferation. Ki-67 is a nuclear antigen, strictly correlates with cell proliferation and is expressed in all phases of the cell cycle except G0. The aim of this study was to investigate the presence of myocyte replication by analyzing the expression of telomerase and Ki-67 in myocytes after myocardial infarction. Method: We studied myocardial samples of twenty hearts with histologic findings of myocardial infarction. Myocardial samples of ten normal hearts were used as controls. An immunohistochemical method was performed with the use of telomerase and Ki-67 antibodies, in order to investigate the expression of these markers in infarcted hearts. Results: The percentage of Ki-67 and telomerase positive expression samples was 85% (17/20 positive samples) and 70% (14/20 positive samples) in myocardium infarction, respectively. Moreover, we report that 70.5% of Ki-67 positive samples expressed telomerase. Positive expression of telomerase and Ki-67 appeared rarely in control group. Conclusions: The increased expression of Ki-67 and telomerase in infarcted myocardium, suggest the presence of myocyte proliferation and this may be a compensatory mechanism that could be replace damaged myocardium.

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PP3-320 IMUNOHISTOCHEMICAL STUDY OF THE ADIPOSE TISSUE IN A FATAL CASE OF ARRHYTHMOGENIC RIGHT VENTRICULAR DYSPLASIA Peter Ghenev1, Ivan Stankulov2, Anton Tonchev3, George Chaldakov3 1 Department of Pathology, Medical University of Varna, Varna, Bulgaria 2 Department of Forensic Medicine, Medical University of Varna, Varna, Bulgaria 3 Division of Cell Biology, Medical University of Varna, Varna, Bulgaria Arrhythmogenic right ventricular dysplasia (ARVD) is a heritable disorder characterized by progressive degeneration and fibrofatty replacement of right ventricular myocardium, causing ventricular tachyarrhythmias and resulting in sudden death at a young age. We report a young man without prior complaints dying suddenly during minor physical effort. The gross autopsy findings in the heart were minimal. The histologic features in both right and left ventricles were typical of ARVD, and consisted of fatty infiltrates with typical cardiomyocyte degeneration of the right ventricle and subepicardial regions of the left ventricle. Several lines of evidence suggest that impairment of cell-to-cell adhesion and involvement of desmosomal proteins may be the underlying pathogenic mechanism in ARVD, via accelerating apoptosis of myocardial cells. As fat replacement is most essential for the diagnosis of ARVD, the aim of the present study is to analyze immunohistochemically ARVD-related adipocytes compared to these from the subcutaneous fat. It is becoming increasingly evident that adipose tissue is a multifunctional organ that produces and secretes multiple factors that can act in both paracrine and endocrine fashion. So it is our opinion that adipose tissue in ARVD is not an innocent bystander and adipocytesecreted cytokines probably play a substantial part in ARVD pathogenesis. PP3-321 ONE STEP CLOSER TO DISCLOSING THE MORPHOLOGICAL SUBSTRAT OF SYNDROM X Tigran Ghevondya National Institute of Health, Armenia Background: The name and existence of several definitions of the syndrome X point to vagueness of this condition. In present research we adhere to opinion, that the ɏ-syndrome is the phenomena of angina pectoris and myocardial ischemia documented by electrocardiography without evidence of stenosis or a spasm of heart coronary arteries. Some researchers explain such pains through spasm of fine arteries of heart. There are no methods to study lifetime morphology of syndrome X at the same patient. We started with the assumption, that among died from first heart attack the number of persons suffered from ɏsyndrome at lifetime should be much more, than among healthy people. The aim was to clarify whether quantitative spatial peculiarities of intramyocardial arterial bed exist at died from the first heart attack? Method: Present research is carried out on 80 hearts received from practically healthy persons died of the violent reasons and on 60 hearts of patients died from acute myocardial infarction. Methods of X-ray micro-angiography of hearts; planimetry of X-ray films of myocardial strips; specimens cutting in three planes; scanning of histological slides under microscope; stereological calculation of volume density of arterial bed and statistics were used. Results: Both X-ray films data and data from histological slides show statistically reliable differences of the arterializations in different parts of a left ventricle wall. It has been revealed almost ninefold difference in mean value of parameters of volume density of arteries calculated for injected hearts in comparison with the data of non-injected

hearts. The wide variation of value of a parameter both in healthy and in hearts with a myocardial infarction is found out. Thus the degree of arterializations in heart muscle with infarction was significantly much lower than arterializations of healthy hearts. The difference was obvious both between the data of X-ray films and data of stereological analysis of injected and non-injected hearts. Conclusion: The types of myocardial angioarchitectonics possessing low volume density are less favorable for heart in respect to blood supply of a myocardium and these types most frequently meet at hearts attacked by acute myocardial infarction. It is possible to assume, that the syndrome X meets more often at persons, who have less favorable angioarchitectonics, which brings to faster ischemia even by week spasms of coronary or small intramuscular arteries. PP3-322 PRIMARY MALIGNANT PERICARDIAL MESOTHELIOMA: A CASE REPORT Selvinaz Ozkara, Ilkay Tosun, Murat Erkan, Fugen Vardar Aker, Pembegul Gunes Department of Pathology, Haydarpasa Numune Education and Research Hospital, Istanbul, Turkey BACKGROUND: Primary malignant pericardial mesothelioma (PMPM) is extremely rare and overall poor prognosis. On reviewing the literature it appears that approximately 72% of all PMPM were actually diagnosed on autopsy. This obviously indicates a silent nature of this highly malignant and lethal tumor. The case of primary pericardial mesothelioma presenting as constrictive pericarditis is reported here, because of rarity. CASE: A 42-year-old male presented with two months history of progressively worsening breathlessness and central chest discomfort on effort. The diagnosis of constrictive pericarditis was made based on CT evidence of pericardial thickening and hemodynamic evidence of constrictive physiology. The history did not reveal any pertinent information such as exposure asbetos fibers. At surgery, the pericardium was thick, white, homogeneous, and very strongly adherent to the myocardium. As a result, resection of the pericardium was incomplete. Postoperatively the patient’s condition deteriorated rather rapidly with low cardiac output state and he died soon after (one month) surgery. No post-mortem was possible. Several hard pericardial samples with irregular shapes were analyzed, the largest measuring 5x5x2cm. Microscopic examination revealed cancer infiltration of the pericardium with a multiple connected nodule morphology. These nodules consisted of large eosinophil cytoplasm, a vesicular nucleus and prominent nucleolus. The cells formed cord-like or tubular structures, confirming the mesothelial origin of the malignancy. Immunohistochemistry was positivity for calretinin, EMA and P53, while staining for desmin was negative. The final diagnosis was “malignant pericardial mesothelioma, epithelial type”. CONCLUSION: Pericardial mesothelioma is extremely rare, although it is the most common primary pericardial tumor. Its incidence was <0.0022% among 500,000 cases in a large necropsy study. Approximately 200 cases have been reported so far and only 25% of these were antemortem diagnoses. PMPM is an aggressive disease and in our patient this was responsible for his death within one month of diagnosis. Herein, we report this rare case of PMPM and present a brief review of the relevant literature. PP3-323 PEDIATRIC CARDIAC SARCOMAS, A SERIES OF 16 NEW CASES WITH LITERATURE REVIEW Allen Burke1, Fabio Tavora2, Irem Ozbudak3, Teri Franks2, Markku Miettinem2 1 CVPath Institute, Inc., Bethesda, MD, USA 2 Armed Forces Institute of Pathology, Washington, DC, USA 3 Akdeniz University School of Medicine, Antalya, Turkey

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Background: Malignant primary cardiac tumors are extremely rare in infants and children, and there are no series published to date. We present a pathological series of pediatric cardiac sarcomas with clinical follow-up. Methods: Pathologic and clinical information from 23 primary heart tumors in patients ” 16 years at presentation were retrospectively reviewed. Chart record and histologic material were available in all cases. Followup information was obtained from contacting health care provider or patient family in 10 patients. A literature review for primary heart sarcomas in children published since 1980 was performed. Results: Of the initial 23 tumors in the series, 7 were reclassified as inflammatory myofibroblastic tumor (IMT) or benign fibrous histiocytoma, based on pathologic features and benign clinical follow-up. Sixteen cases remained as primary heart sarcomas. There were 14 males and 2 females, mean age of 10.4 years (range 0-16). Clinical presentation was variable and included recurrent pericardial effusion (1), respiratory distress (2), pulmonary emboli (1), shortness of breath (5), arrhythmia (2), chest pain (4), congestive heart failure (1) and constrictive pericarditis. Six patients presented with tumor in the left atrium, 2 in the left ventricle, 4 in the right atrium, 2 in the right ventricle, 2 in the pericardium and 1 in the mitral valve. Histologically, there were 5 leiomyosarcomas, 3 undifferentiated round cell sarcomas, 2 pleomorphic undifferentiated sarcomas, 3 angiosarcomas, 2 rhabdomyosarcomas, 1 osteosarcoma, and 1 fibrosarcoma. The youngest patient with angiosarcoma was 14 years, the youngest patient with pleomorphic sarcoma 12 years, and the ages at presentation of leio-, rhabdo- and undifferentiated round cell sarcomas ranged from 1 – 16 years. Three patients presented with metastatic disease, all with lung metastases. In cases with follow-up, mean survival was 8.7 months; all patients were dead of disease, with one alive with recurrence. Of the review of 12 published reports, 4 were considered likely IMT, based on histologic description and benign follow-up. The remaining 8 were angiosarcomas (2), rhabdomyosarcomas (3) liposarcoma (1), fibrosarcoma (1) and sarcoma, not otherwise specified (1). Prognosis was uniformly fatal, with survival up to 13 months. Conclusions: Primary pediatric sarcomas of the heart have a dismal prognosis, are of varied histologic types, have a male predominance, occur most frequently in the atria, and should be clearly separated from cardiac IMT. PP3-324 INFLAMMATORY PSEUDOTUMOR OF THE HEART: A CASE REPORT Chtourou Imen, Bahri Zouari Ibtissem, Chaari Chiraz, Zribi JihèNe, Gouiaa Naourez, Makni Saloua, Sellami Boudawara Tahya Pathology Department, Habib Bourguiba Hospital, Sfax, Tunisia Introduction: Inflammatory pseudotumor is a rare entity which most commonly involves the lung and the orbit. It rarely occurs in the heart: only 14 cases are reported in the literature mainly in children and adolescents. In this report, we review the relevant literature and discuss the pathogenesis, clinical manifestation, the diagnosis and the treatment of cardiac inflammatory pseudotumor. Case report: A 31 year old woman presented since seven days a fever which was followed by chest pain. The clinical examination was normal. Viral serologies and the assessment immunological were normal. Radiological explorations (ultrasonography, thoracoabdominal tomography and magnetic resonance imaging) revealed the presence of a cardiac mass which measured 24x13mm suggesting a thrombus. Anti coagulant treatement was administrated, but no improvement was observed. Consequently, the patient was operated. Histological examination of the mass showed spindle shaped cells with eosinophilic cytoplasm and elongated bland nuclei; these cells were accompanied by a variable number of inflammatory cells including lymphocytes, macrophages, plasma cells and eosinophils. En immunohistochemical stain, the spindle

cells expressed actin and vimentin, but were negative for keratin, S-100 protein, CD34 and CD1a. Basing of this finding, the diagnosis of inflammatory pseudotumour of the heart was made. Conclusion: Inflammatory pseudotumour of the heart is rare, occurs mainly in children. Clinical presentation is variable. Tumor location and its characteristics can be observed by radiological findings but histological examination is necessary to make diagnosis. Complete surgical resection is the treatment of choice. PP3-325 UNUSUAL CARDIAC TUMORS OTHER THAN MYXOMA. PRESENTATION OF 2 CASES Ahu Senem Demiroz1, Cem Leblebici2, Buge Oz1, Sergulen Dervisoglu1, Feriha Oz1 1 Department of Pathology, University of Istanbul, Cerrahpasa Faculty of Medicine, Istanbul, Turkey 2 Department of Pathology, Istanbul Education and Reseach Hospital, Istanbul, Turkey Background: The presentation of heart tumors is very rare. About 75% of primary tumors are benign, and 75% of these are atrial myxomas. Malignancy is found in fewer than half of primary tumors of the heart. The majority (95%) are sarcomas, primarily angiosarcomas and undifferentiated sarcomas. Primary cardiac tumors are high-grade sarcomas with a high metastatic potency that often becomes evident early after surgery.The presence of metastatic tumor to the heart usually indicates widespread metastases. Lung carcinomas are the most commonly encountered tumor followed by breast and pancreas cancer and melanoma. We present 2 cases with rare heart tumors Case 1: 24 year old woman presented with cardiac arrhythmia and was found to have a mass on mitral valve and left atrium. Histopathology of the resected specimen was a high grade osteosarcoma composed of fibroblastic, chondroblastic and osteoblastic components. An immunohistochemical study was performed by using S-100 antigen to differentiate the osteosarcoma from malignant peripheral nerve sheath tumor. There was positive staining in histiocytes but negative staining for tumor cells. So the histological and immunohistochemical findings of the permanent sections of the mass were consistent with primary cardiac osteosarcoma. Case2: 44 year old man presented with chest pain. His radiological findings revealed a mass in right atrium. Histopathology of the incisional biopsy was an undifferentiated malignant tumor with partial epitheloid pattern, showed extensive necrosis and invasion through pericardium and myocardium. Metastatic undifferentiated carcinoma was taken into differential diagnosis, but keratin was negative and no suspicious primary focus was found. Immunohistochemical results didn’t support the relatively common primary malignant cardiac tumors such as rhabdomyosarcoma and angiosarcoma. Malignant paragangiioma was excluded due to chromogranin negativity and malignant melanoma was excluded due to HMB 45 and Mart-1 negativity. Conclusion: Sarcomas as unusual primary cardiac tumors should be taken in differential diagnosis when cardiac myxoma and metastatic carcinomas are excluded. PP3-326 THE RELEVANCE OF CO-MORBIDITIES FOR IN HOSPITAL LETHALITY OF PATIENTS WITH ACUTE MYOCARDIAL INFARCTION: CLINICOMORPHOLOGICAL STUDY Eleonora Vataman1, Vladimir Vataman2 1 Institute of Cardiology, Chisinau, Republic of Moldova 2 Medical and Pharmaceutical University, Chisinau, Republic of Moldova Aim: To assess the impact of co-morbidities on myocardial infarction outcomes during acute phase of the disease. Methods: The survey vas based on retrospective analyze of necropsy

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protocols for a 10-years period. 1269 cases of in-hospital death of patients (mean age 67,9±0,55 years) with myocardial infarction were studied. Statistical analysis included clinical features of myocardial infarction and its complications; co-morbidity; results of morphologic study. The aim of statistical analysis was to determine the factors which might have crucial role for the evolution of myocardial infarction and which could be used for predicting some fatal complications: acute ventricular aneurysm, cardiac rupture, pulmonary oedema, acute cerebrovascular accident, etc. Results: The group included 698 (55%) men and 571 (45%) women. Mean hospitalization period was 10,9±0,88 days. The frequency of co-morbidities were: arterial hypertension (27,1%); diabetes mellitus (21,2%), pulmonary diseases (11,5%), kidney pathology (11,5%), hepatitis/cirrhosis (3,7%), gastrointestinal pathology (4,2%), obesity (7,6%). Causes of death were: pulmonary oedema – 40,3%, acute heart failure – 29,7, cardiogenic shock – 19%, sudden death- 14%, left ventricular parietal wall rupture – 12,2%. Other important complications: rhythm disturbances (chronic atrial fibrillation – 12,7%, high gradation extrasystoles- 4,6 %), stable conduction abnormalities (atrio-ventricular block – 2,36%, complete left bundle block – 2,36%), development of left ventricular endocavitary thrombosis (6,7%), periinfarctic pericarditis (1,73%). Among clinical parameters, responsible for the development of myocardial infarction complications and death, discriminant analysis revealed myocardial infarction characteristics: depth (transmural – 33,9%), location (46,3% - anterior, 30% - posterior), width (anterior extended – 27,5%, circular – 23,5%, posterior extended – 19%), evolution (repeated – 39,5%, recidivant – 13,5%), presence of left ventricular aneurysm (total – 12%, acute – 4,4%, chronic – 8,2%). Co-morbidities were not found as specific determinants of lethal outcomes in acute phase of myocardial infarction. PP3-327 DYNAMICS OF ADAPTABLE RESERVES AT THE PATIENTS SUFFERING FROM ISCHEMIC HEART DISEASE AGAINST THE BACKGROUND OF TREATMENT WITH CITOPROTECTOR BIOR Lina Remish1, Ivan Butorov2, Vladimir Remish3, Vasile Anestiade3 1 Academy of Sciences of Republic Moldova, Republic of Moldova 2 Medical State University of Republic Moldova, Republic of Moldova 3 Scientific Centre of Pathobiology and Pathology of the Academy of Sciences of Republic Moldova, Republic of Moldova Both pharmacological means and the program for improvement of quality of a life and increase of tolerance to physical load of the patient with IHD should be used reasonably, render positive influence on the basic links of pathogenesis of the illnesses and increase nonspecific resistance of an organism. The purpose of research - to study the dynamics of adaptable reserves of the cardiovascular system at the patients with the ischemic heart disease during treatment. Material and methods of research. 135 patients with ischemic heart disease from which 21 (15,5%) had stenocardia I CF, 98 (72,6%) – a stenocardia II CF and 16 (11,9%) – a stenocardia III CF were under supervision. The most frequent risk factors were: an arterial hypertension -78 the patients (57,8%), hypercholesterolemia - 82 (60,7%). All patients received traditional medicamentous treatment, 82 patients (the basic group) in addition received cytoprotector BioR by 1, 0 ml intramuscularly daily during 20 days. Results of research. The analysis of dynamics of the clinical symptoms and syndromes against the background of the carried out therapy has shown, that in patients of the basic group pains in heart, a short breath, instability of arterial pressure decreased for 6-7 days earlier, than in group of comparison. It is established, that in patients with

initial depression of processes of tissue respiration the increase of the constant of the speed oxygen absorption (from 0,032±0,003 up to 0,040±0,002 sec, ɪ <0,001), alongside with the reduction of the exhaustion time of oxygen stocks (from 155,2±10,8 up to 131,7±11,2 sec, ɪ <0,001), and after treatment time of exhaustion of half of oxygen stocks (from 39,6±1,2 up to 32,7±1,6 sec, ɪ <0,001) has been revealed. That demonstrates the improvement of processes of recycling of oxygen by the tissue. In patients of the control group positive changes have also been revealed, but they are less expressed and are statistically doubtful. The improvement and increase of compensative-adaptive possibilities of the organism during application BioR are also testified by the positive changes of the hemodynamics parameters in the patients. So, the heart index has increased with 1,41 ± 0,03 up to 1.56±0,04 l/mines/m², (ɪ <0,001), while in group of the control with 1,43 ± 0,05 up to 1.48±0,06 l/mines/m², (p > 0,01); the Volume of performed work in the patients of the basic group has increased by 32,7%, in group of the control – by 6,7%; tolerance to loading has increased by 19,8% (ɪ <0,001) in the patients of the basic group, and only by 5,85% (p> 0,01) in control group. The involvement of cytoprotector BioR in the complex therapy of patients with IHD promotes the increase of the compensativeadaptive reactions of an organism and clinical efficiency of the treatment. PP3-328 SUDDEN DEATH FROM TUBERCULOUS MYOCARDITIS Butcovan Doina, Tinica Grigore, Grigoriu Carmen Cardiology Center Iasi, Romania Tuberculosis is an important public health problem considering to spare four organs: heart, skeletal muscle, thyroid and pancreas. We present a case of tuberculous myocarditis diagnosed on a post-mortem cardiac biopsy to a girl patient of fifteen years old, presented with unknown history of tuberculosis, but having a juvenile rheumatoid arthritis for who received corticosteroids for a long period of time. Clinical evaluation revealed rhythm disturbances on electrocardiogram, right cardiac failure and right upper lobe consolidation on chest X-ray. Microscopical examination of necroptic biopsies evidentiated specific lung giant-epithelioid granulomas with caseous necrosis and extensive infiltrative tuberculous lesions on the right lung associated with myocardial epithelioid granulomas without evident caseous necrosis. Tuberculous etiology was confirmed by applying a specific stain for Mycobacterium tuberculosis, obtaining a positive Ziehl-Neelsen result. We hypothesize that the mechanism of death was severe ventricular arrhythmia due to granulomatous proliferation in the structures of the interventricular septum. Because the involvement of myocardium in tuberculosis is rare, the increasing recognition of the entity and the use of endomyocardial biopsy may help us for detecting of more cases of this "curable" form of cardiopathy. PP3-329 CARDIOVASCULAR CAUSES OF SUDDEN UNEXPECTED DEATH; SIX YEARS PROCESS IN COUNCIL OF FORENSIC MEDICINE Arzu Akcay Turan1, Ercument Aksoy2, Oguzhan Melez3, Oguzhan Ekizoglu4 1 Council of Forensic Medicine, Istanbul, Turkey 2 University of Marmara, Faculty of Medicine, Department of Forensic Medicine, Istanbul, Turkey 3 Council of Forensic Medicine, Istanbul, Turkey 4 University of Istanbul, Faculty of Medicine, Department of Forensic Medicine, Istanbul, Turkey Background: Cardiovascular deaths constitute the major cause of sudden and unexpected deaths. Aim: To evaluate the pathological changes of 1424 cases reported as cardiovascular deaths,

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autopsied in Council of Forensic Medicine in Istanbul, Turkey Methods: Autopsy reports of sudden unexpected deaths of cardiovascular origin between the years of 1999-2001 were retrospectively analyzed. In the sudden unexpected cardiovascular originated deaths between 2002 and 2004 whole hearts were sampled and examined systematically after being fixed. Findings of these two periods were compared in order to the role of cardiac sampling and systematic evaluation in the diagnosis of sudden unexpected deaths. Results: 1424 autopsy cases were analyzed. 1240 (87,1%) of these cases were male and their age range was 56± 15, 184 (12,9%) were female and the age range was 49± 20. 657 (46%) of these cases were died suddenly, 619 (43,5%) were found death and 98(6,9%) were died at the hospital. We found chronic ischemic changes in 724 (50,8%), acute infarction in 177 (12,4%), valvular disease in 44 (3,1%), myocarditis in 21 (1,5%), cardiomyopathies in 7 (0,5%) and left ventricular hypertrophy in 326 (22,9%) cases and the rupture of an aortic aneurysm was the cause of death in 32 (2,2%) cases and 79 (5,5%) cases were morphologically negative. When compared to the group of only myocardial sections are carried out at autopsy, the incidence of cardiomyopathies (7,8%), congenital heart diseases (3,1%) and acute infarction (14,1%) was detected to be high in the group where heart were whole sampled. Discussion: Pathological changes detected during the autopsy in a systematic manner will not only support the clarification of the cause of death, but also provide in depth knowledge about sudden and unexpected deaths. PP3-330 DETECTION OF APOPTOSIS IN ENDOMYOCARDIAL BIOPSY SPECIMENS FROM PATIENTS WITH DILATED CARDIOMYOPATHY Sofija Glumac1, Anna Batistatou2, Petar Otaševic3, Zoran Popovic1, Zorica Stojsic1, Aleksandar Neskovic4, Jovan D. Vasiljevic1, Niki Agnantis2 1 Institute of Pathology, Medical School, Belgrade, Serbia 2 Institute of Pathology, Medical School, Ioannina, Greece 3 Institute for CV Desease Dedinje, Belgrade, Serbia 4 Cinical Center Zemun, Serbia Background: Apotposis is a regulated energy-consuming process and physiologically found in embryogenesis and the maturation of multiple cell systems, and pathologically in various diseases. Apoptotic cardiomyocyte death is encountered in various heart diseases such as DCM, myocarditis, coronary artery disease, reperfusion injury, as well as right ventricular dysplasia, and hypertensive heart disease. The most widely used method to identify apoptosis in human histological material is the TUNEL assay. Design: The aim of the present multicentric study was to asses the apoptotic index by TUNEL method in endomyocardial biopsies from patients with DCM. Apoptosis was detected by using the TUNEL method in 33 patients. The values of apoptotic index were expressed as means with standard deviations. Biopsies were performed at the Instutute of Cardiovascular Diseases Dedinje, Belgrade. Results: The mean age of patients with dilated cardiomyopathy (DCM) was 29 males and 4 females. The mean age of patients was 44 ( 31-59 years). All cases of DCM were diagnosed both clinically and pathologically DCM was detected in 26 cases (79%). In this group of patients, the most common diagnosis was advanced DCM (9 cases or 35 %), followed by mild DCM (7 cases or 27 %), early stage of DCM (3 cases or 11%); recovery phase (6 cases or 23%) and hereditary CM (1 case; 4%). In 3 cases (9 %) Idiopatic dilated cardiomyopathy was found and the most common diagnosis was early DCM (1 cases;33 %), mild DCM (1 case; 33%) and focal myocarditis (1 case; 33%).DCM and insuffitientio valvulae mitralis were found in 4 cases (12 %) and in this cases pathohistological diagnosis were: early DCM (2 cases; 50 %); advanced DCM (2 cases; 50 %). TUNEL-positive myocytes were found in 26 of 33 cases with DCM (78,8%). The apoptotic index ranged 1,4 - 10,4%. The mean apoptotic index for the whole

analyzed group of DCM was 3,3 ± 2,485%. Conclusion: The present study focused on the morphological analysis of apoptosis in DCM in endomyocardial biopsies specimens. We have found relatively high percentage of apoptosis in our series, in 78.8% of cases, as well as relatively high apoptotic index. These results could be explained by manifested cardiac failure of our patients, and represent midle values in other reported series.