key: cord-0041555-6yecun89
authors: nan
title: Poster Session
date: 2009-08-31
journal: Cytopathology
DOI: 10.1111/j.1365-2303.2009.00691.x
sha: d74d5bf5e583be773f05cb01c09f36520e50e3cb
doc_id: 41555
cord_uid: 6yecun89

nan

origin of the tumor. It is good to notice that in some cases of highrisk endometrial adenocarcinomas p16 positivity can be found too. Objective: In our Laboratory the Shandon Cytoblock Ò technique is used to optimize cytological diagnoses in difficult cases. In this case of atypical glandular cells (AGC) we used the residual material of the ThinPrep Ò (Hologic) specimen for cytohistology and immunohistochemistry. Materials and Methods: In 2008 we received a ThinPrep slide of a 50-year-old woman without complaints, containing many thick cell groups, that were impossible to classify. The vial still containing the brush (Cervex-Brush Ò Combi) was therefore placed in a shaker for ten minutes to dislodge the material trapped between the bristles. The residual sample was used to prepare paraffin sections. From the block 4 lm sections were cut and stained with Papanicolaou and the biomarkers Ki-67 (proliferation marker) and p16INK4a (HPV indication) . Results: In the paraffin sections we encountered numerous small tissue fragments covered with slight atypical glandular cells intermingled with many goblet cells. Since we could not detect adenocarcinomatous growth, the case was signed out as adenocarcinoma in situ (AIS) of the intestinal type, with positive staining for Ki-67 and p16INK4a. This diagnosis was confirmed by histology (cone biopsy). Conclusion: Cytohistology and immunohistochemistry on the residual of cervical ThinPrep specimens are very helpful in cases with thick cell groups. In this case of AIS of the intestinal type, where no feathering or rosettes were present in the ThinPrep specimen, cytohistology could avoid a false negative diagnosis. Objective: In the near future, it will become completely out of date to ask cytotechnologists to screen a complete cytology slide (containing over 200 000 cells) in search of abnormal cells. Computer-assisted screening is not only much more fun and relaxing, but also fully profits from the skills of the cytotechnologists, because they can focus and spend ample time in judging the few microscopic fields selected by the computer. To analyze the effects on the cytoscores for computer-assisted screening when a large throughput laboratory switches from neural network scanning (PAPNET) dating back to the eighties to the state of art Imager (Hologic). Materials and Methods: Prior to July 2007, 57 913 ThinPrep slides were scanned via neural networks and the year after 64 651 with the Imager. 99 705 concerned populations screening of asymptomatic women, the remainder were indication cases. Results: Positive cytoscores for ‡ CIN III were for neural networks 0.28 and 0.61, for the Imager 0.44 and 0.87, with higher values in the populations screened women than in the indication cases. The same trends were observed for the lesser lesions and for the cytoscores for the infections. Conclusion: The highest positive scores were realized for the combination indication cytology and Imager scanning. The switch to the Imager has enhanced our diagnostic acumen over the full range of cytology in our practice of computer-assisted screening. Objective: My objective was to reflect on the factor joy in screening for a Japanese cytotechnologist, in a study comparing thin layer and conventional cancer slides. Materials and Methods: Out of the LCPL archives, 10 ThinPrep Ò slides and 10 conventional slides were retrieved to assess the similarities and differences between the two preparation methods. The focus was morphologic features as such taught by the Brazilian cytotechnologist N. Meyer-Alcâ ntara. Results: For the thin layer slides cell size, fixation, background features, uniformity of staining, clearness of nuclear and cytoplasmic detail were studied. For the conventional smear cell size, fixation, smear pattern, and background features were analyzed. Conclusion: In the thin layer slides, fixation was superior but rounding up of the cancer cells was more pronounced which might be a problem. However, in all cases there were single more flattened cancer cells that could be studied in detail. The background features differed in the conventional smears and the fixation was sometimes poor. Moreover, the cells were flatter and larger. Finally, I have to mention that I learned to enjoy screening here in Holland, especially of the ThinPrep slides. Brazilian joy, as I have encountered in Holland, has proven to be very important for me.

were screened using neural network scanning (NNS) and 24 866 via the Imager. In the NNS series there were 958 consultation cases and in the Imager screening (IS) 1138. For each cytotechnologist (CT) the percentage of full rescreen slides at the Imager was calculated, that is of cases diagnosed as negative based on the Imagerselected 22 fields of view (FOV) . Results: The percentage of full rescreen Imager slides varied per cytotechnologist (3) (4) (5) (6) (7) (8) (9) (10) (11) (12) (13) (14) (15) (16) (17) (18) and decreased for each cytotechnologist from July until December 2007.The percentage of consultation cases, that is those cases that the CT did not dare to sign out as negative on his/her own, varied for each individual, junior CTs having more consultations than senior CTs. The cases selected for consultation by the junior CTs concerned mainly slides finally signed out as negative. The consultations of the senior CTs consisted mostly out of ‡ CIN 2 cases. Conclusions: For both types of computer-assisted screening (NNS and IS) experienced cytotechnologists were more confident in signing out negative cases than junior CTs. Objective: To compare the diagnoses obtained by cytology and biopsy with the results of the detection and genotyping of HPV, associating the most frequently types in different diagnoses. Materials and Methods: A retrospective study was performed, researching the results of 218 liquid-based Pap smear classified as NIL, ASC-US, ASC-H; SIL, HSIL and LSIL, of which 70 cases had biopsy. Results of biopsies in which no viral cytopathic effect was observed were considered negative. The other results were classified as LSIL, HSIL and condyloma.After the Pap smear result, the samples were analyzed by the Human Papillomavirus Clinical Arrays â for detection and genotyping of 35 HPV with the greatest clinical importance. The results were statistically analyzed by Chi-Square Test and the analysis of frequencies.

Results: Statistical analysis by Chi-Square Test shows a significant association between diagnosis and outcome of cytologic genotyping of HPV. The more relevant results are the cytological diagnosis NIL (with 73.2% negative for HPV), the diagnosis LSIL (with 96.7% of positivity for HPV) and the diagnosis of ASC-US (with 60% of positivity for HPV). There is also a significant association between the histological and cytological diagnosis. It was found that 91.3% of the histological diagnosis was consistent with the cytological diagnosis of NIL. In 45.5% of the cytologic diagnosis of ASC-US, the histological diagnosis did not identify viral cytopathic effect and in 45.5% of cases were diagnosed LSIL. Concerning the cytological diagnosis of LSIL, in 66.7% of cases was not identified viral effect on histological examination. The analysis of frequencies shows the presence of HPV 16 in 21.4% of diagnoses of NIL and in 26.7% of diagnoses of ASC-US. The HPV 53, of probable high-risk, is frequent both in cytological diagnosis of NIL as ASC-US, LSIL and HSIL.

The inexistence of a 100% concordance between the cytologic diagnosis, histology and the results obtained with the HPV DNA tests, reports to the importance of association the cytology with techniques of detection and genotyping HPV, to increase the reliability of screening for cervix cancer. Objective: Compare three stain protocols (A -Papanicolaou Standard, B -Papanicolaou Alternative, C -Papanicolaou Quick Ò ) so as to determine speed, economy and stain quality. Methods: Fifty samples of gynaecologic cytology were processed in ThinPrep Ò . Three slides from each sample were obtained with each being submitted to a different Papanicolaou staining (A, B, C). The stain's quality was evaluated by four observers at three different times, focusing on the cellular preservation, acidophil and basophilic structure, nuclear basement, cytoplasm differentiation, stain quality and processing artefacts. Statistic tests were performed: One-way ANOVA to verify if there were differences between the quality of the three Papanicolaou stainings and; ANOVA repeated measures to verify if there were significant changes in the quality of the stained slides over the storage time. At the same time, the cost and the execution time for the respective protocol was determined. Results: Differences were not verified in the final quality of the three protocols. Changes were observed in the staining quality over the storage time for all protocols. Alternative Papanicolaou B is faster and more economical than the others. Conclusion: It was ascertained that Alternative staining protocol is faster and more economical than Papanicolaou standard and Papanicolaou Quick Ò , which provides an equivalent in terms of quality, is a less expensive method and also has environmental advantages. Objective: 'Projeto Expediçõ es Cientificas e Assistenciais' is an annual project which started in 2004, created by the medical students from the Medical School of 'Santa Casa de Sã o Paulo'. The project is organized by third year medical students, doctors, nurses and members from the faculty. This year, in its fifth edition, the project took place in Areia Branca, a less favored rural district belonging to the city of Itapeva in the interior of Sao Paulo, Brazil, with a population of 5052 inhabitants, its main economic activity being tomato cultivation. The project took place between the 24th and 29th January 2009, where 231 professionals and students were involved. It aims to provide a comprehensive and multi-disciplinary approach, with emphasis given to cervical cancer prevention, and intervention in primary medicine to a population with poor access to health care. Materials and Methods: A total of 2000 patients were seen by specialists and 71 women were screened for cervical cancer through conventional Pap smear. Cases were reviewed by a cytopathologist and diagnosis was made according to the Bethesda System, 2001. Results: Age interval was from 16 to 76 years, showing 2 HSIL, 5 LSIL, 2 ASC-US and 62 inflammatory conditions. Microbiological studies showed 2 infections by Trichomonas vaginalis, two Candida albicans, 12 Gardnerella vaginalis, 18 mixed flora and the remaining cases corresponding to lactobacillus. Conclusion: The finding of nine positive cases (12.5%) is a great indicator for the need of an early screening of this less favored population. This approach also reinforced the students' commitment towards a holistic view of the patient, aiming the primary prevention and outcome of the patients. Background: As suggested by the nomenclature, a diagnosis of AGC on cervical cytology samples strongly suggests underlying glandular lesions of either endocervical or endometrial origin. These patients are also at increased risk for concurrent squamous dysplasias. In this study, the histologic outcome of patients diagnosed with AGC on ThinPrep Ò (TP) at a single institution over a 6.3-year period was determined and differences in histologic outcome were stratified by patient age. Design: A computerized search of the cytology laboratory information system was performed for all TP cervical cytology samples diagnosed as AGC from March 2002 through July 2008. The histologic outcome of the AGC cases was divided into eight Categories: no histologic follow-up, negative, microglandular hyperplasia, complex hyperplasia, low-grade dysplasia (LGD), high-grade dysplasia (HGD), benign endometrial polyp (EMP), adenocarcinoma in situ (AIS), and malignancy. EMP, squamous dysplasia, AIS, and malignancy were considered to be clinically significant lesions. The data was stratified by patient age into two groups; patients 35 years of age. Results: Of 107 cytology cases diagnosed as AGC on TP, 63 patients (59%) had follow-up biopsies and 34 cases (32%) had clinically significant lesions. Of the clinically significant lesions, 32 (94%) occurred in women greater than 35 years of age and only two cases (6%) occurred in women aged 35 years or less. All malignancies occurred in the older age group. AIS accounted for all significant lesions in women aged 35 or less. The results are summarized in the table below. Conclusions: At our institution, clinical recommendations for follow-up biopsy of TP AGC were ignored in 41% of cases. The histologic diagnostic yield for significant abnormalities following TP AGC diagnosis was only 32% mostly occurring in the older (> 35) age group. Of these clinically significant lesions, 50% were histologically confirmed as definitive malignancies (15 endometrial primaries and two metastases), strongly suggesting that malignancies are underdiagnosed in our laboratory. Our results reflect the importance of adhering to established clinical management guidelines for patients diagnosed with AGC. Objective: Prophylactic administration of a quadrivalent HPV6/11/ 16/18 vaccine is highly effective in preventing HPV6/11/16/18-related CIN and external genital lesions (EGL = VIN, VaIN, and genital warts). We report the impact of the vaccine on the incidence of all HPV disease, regardless of causal HPV type. Materials and Methods: 17 622 women aged 16-26 were enrolled in one of two randomized, placebo-controlled trials. Vaccine or placebo was given at Day 1, Month 2 and 6. Pap testing occurred at Day 1 and every 6-12 months for up to 48 months. Colposcopy referral was algorithm-based. We estimated number of cases prevented annually per 10 000 vaccinated women, in terms of risk difference (RD) or prevented risk based on subtracting the rate in the vaccine arm from the rate in the placebo arm in: (1) an unexposed population that approximates sexually naïve females;

(2) a mixed population of HPV-exposed and unexposed women which approximates the general population of sexually active women; and (3) the population of women already exposed to HPV. Results: Within~3.6 years, we observed reductions in the number of lesions in all populations. In the unexposed population (defined as DNA negative at baseline to all of 14 HPV types, seronegative to HPV6/11/16 and 18, and with a normal Day 1 Pap), the annual estimated number of cases prevented per 10 000 vaccinated women was: any CIN = 71 (RD = 0.71.95%CI: 0.4, 1.0); any CIN2/3 or AIS = 35 (RD = 0.35.95%CI: 0.2, 0.5); and any EGL = 102 (RD = 1.02.95%CI: 0.8, 1.2). In women who were exposed to HPV (defined as DNA positive at baseline to any of 14 HPV types and/or seropositive to HPV6/11/16 or 18), the estimated number of cases prevented was similar to the prophylactic population: any CIN = 99 (RD = 0.99.95%CI: 0.4, 1.6); any CIN2/3 or AIS = 34 (RD = 0.34.95%CI: < 0, 0.75); any EGL = 60 (RD = 0.6.95%CI: 0.3, 0.9). Conclusions: Our data suggest that whether we offer HPV vaccination to a population of women who are HPV naïve, a mixed population of naïve and exposed (data not shown), or a population of previously exposed women, we could expect to have similar public health impacts in terms of disease reduction in the years immediately following vaccination. Seven cytology laboratories all over Estonia is currently involved in screening programme. The project is financed by health insurance fund that is inviting women to attend for cervical screening on a five-yearly basis. In 2008 women who were born in 1948, 1953, 1958, 1963, 1968 and 1973 received the invitation letters. 1898 women participated East-Tallinn Central Hospital in 2008. As a result of this programme we discovered three early stage cervical adenocarcinomas.

First case: A 35-year old woman recieved the PAP smear diagnosis of AGUS, suspicion for adenocarcinoma in situ (AIS). HR-HPV type 18 were detected by HPV DNA analysis. Cervical conization was performed and histological diagnosis of the cone biopsy material was adenocarcinoma in situ. Immunohistochemistry for p16INK4a revealed strong positive staining in glandular epithelial cells.

Second case: 40-year old woman recieved the PAP smear diagnosis of AGUS, suspicion for adenocarcinoma in situ. HR-HPV type 16 was detected by HPV DNA analysis. Cervical biopsy was performed, but dysplasia was not seen in biopsy material and repeat PAP test in 6 months was recommended. In a month a conization was still performed and histological diagnosis of cone biopsy material was adenocarcinoma in situ (AIS). Immunohistochemistry for p16INK4a revealed strong positive staining in glandular epithelial cells.

Third case: 45-year old woman recieved the PAP smear diagnosis of AGUS, suspicion for adenocarcinoma in situ. HR-HPV type 16 was detected by HPV DNA analysis.Conization was performed and histological diagnosis of the cone biopsy material was adenocarcinoma in situ (AIS) and CIN I. Immunohistochemistry for p16INK4a revealed strong positive staining in glandular epithelial cells. Aim: To determine the rate of false-negative cervical cytology findings using the methods of internal quality control. Materials and Methods: During a 3-year period, 83 547 conventional Pap smears were analyzed. Out of 40 000 negative cytology findings on initial screening, 1033 (2.59%) Pap smears were submitted to random rescreening, whereas 23.72% (10 200/43 547) Pap tests initially evaluated as inflammatory or reactive lesions of squamous epithelial cells were separated for control cytology and rescreened by senior cytologist. Employing the method of random sample, the cytologist analyzed 693 (6.79%) of these 10 200 Pap smears. Results: The initially negative screening findings were confirmed in 97.39% (1006/1033) Pap smears, whereas 2.61% (27/1033) proved to be false-negative findings including ASCU-S (1.65%), ASC-H (0.29%), H-SIL (CIN III) (0.10%) and AGC -probably reactive lesions of endocervical origin (0.58%). Negative rescreening performed by senior cytologist was confirmed in 85.43% (592/693) of Pap tests, whereas 14.57% (101/693) proved to be false-negative findings including ASCU-S (12.84%), LSIL (CIN I) (0.58%) and AGC -probably reactive lesions of endocervical origin (1.15%). No case of HSIL was recorded in this group. Conclusion: Employing the methods of internal quality control on initially negative Pap smears upgrades not only the sensitivity of cytologic screening but also the quality of cytotechnologist performance.

Objective: DNA methylation is an early event in carcinogenesis and is often present in the precursor lesions of various cancers. Testing for DNA methylation has potential in cancer screening. The aim of this study was to investigate the feasibility of detecting methylated DNA as a screening tool for squamous cell carcinomas (SCC) and squamous intraepithelial lesions (SIL) in cervical scrapings. Materials and Method: A multiplex, nested, methylation-specific polymerase chain reaction approach was used to examine promoter methylation of 12 genes in biopsy-proven SCC (n = 69), high-grade SIL (HSIL, n = 67), low-grade SIL (LSIL, n = 32), and negative cases (n = 41) from liquid-based cytology samples.

Results: For eight genes (DAPK, HIC-1, HIN-1, MGMT, RAR-b, RASSF1A, SHP-1, and Twist), the frequency of methylation increased statistically significantly with increasing severity of the cervical squamous lesions. The number of methylated genes increased with the severity of cervical squamous lesions (P < 0.001). Clinical stages and the status of lymph node metastasis of SCC had no effect on methylation frequency. At least one of the three genes (MGMT, RAR-b, and Twist) was methylated in 78.7% of samples with HSIL/SCC but in only 17.8% of samples with LSIL or less. The estimated specificity of the three-gene panel was 82.2%, and its sensitivity was 78.7% for detecting HSIL/SCC. Conclusion: Although aberrant DNA methylation has the potential to function as a molecular biomarker of HSIL and SCC in liquidbased Pap tests, additional genes that are selectively methylated in HSIL and SCC are needed to improve the clinical performance. Objective: Transitional cell carcinoma of the cervix is a recognized clinicopathological entity with a spectrum of papillary patterns ranging from squamous morphology to pure transitional differentiation. However cytologic features are far less documented, especially in liquid based sampling. We report the morphology observed in BD-SurePathTM liquid based technology. Case Report: A 49 year old female complaining of exacerbated diffuse abdominal pain is investigated in the emergency unit. Colposcopy reveals a bleeding papillary exophytic lesion of the cervix; a cytologic sample and a day afterwards a biopsy are sent to the laboratory. CTscan demonstrating invasion, the patient is staged FIGO IIb and treated by radiotherapy. Results: The cytology was compatible with an epidermoid carcinoma. The biopsy enhanced the papillary architecture covered with basaloid transitional cell-like cells. Immunostainings showed a CK7+/CK20-profile with p16 positivity. Ki67 and p63 were strongly and diffusely expressed. Invasion could not be evaluated on this superficial biopsy. The cytological characteristics are poorly clearcut. The cellularity appears increased in a dirty background like in common squamous and glandular carcinomas. Most of the cells are loosely dispersed or aligned in somewhat papillary clusters lacking obvious features of HSIL or glandular differentiation. The cells are elongated, fusiform without polarization. The high mitotic activity and the nuclear patterns exclude papillary immature metaplasia (PIM). Koilocytosis and keratinisation are not found despite a positive HPV-DNA testing by Hybrid Capture II performed on the cell pellet. Conclusion: The characteristic cytologic features of this rare variant of squamous carcinoma might be more easily recognized if more comprehensively illustrated through further case reports and in traditional textbooks.

high risk types. Thus HPV genotyping is important to identify patients for colposcopy when cytology is ambiguous. Since each HPV type has different malignant potential and coinfection with multiple hrHPV types is associated with worse prognosis, information of the precise HPV type in LSIL is important for clinicians. Conclusions: We conclude that HPV genotyping is an appropriate tool in clarifying abnormal cytology and it can differentiate lesions that may progress and require early treatment. The aim of this research was to evaluate the distribution of Pap smears of pre-neoplasic and neoplasic lesions, by age groups, which is part of the routine of the Division Pathology of Adolfo Lutz Institute. Retrospective analysis of diagnoses derived from women of the public health system carried out between [2004] [2005] [2006] [2007] [2008] . Of the 222 025 samples analysed, 1.67% (3674) were classified as unsatisfactory, 92.98% (206 439) negative and 5.36% (11 912) with abnormal cytology diagnoses. Of those cases with abnormal cytology, 6,647 (55.80%) were for atypical squamous cells of undetermined significance (ASC-US), 575 (4.83%) for atypical glandular cells of undetermined significance (AGC), 3,264 (27.40%) low-grade squamous intraepithelial lesions (LSIL), high-grade squamous intraepithelial lesions (HSIL), 7 (0.06%) adenocarcinoma in situ, 8 (0.07%) adenocarcinoma,131(1.10%) invasive carcinoma(CA). The distribution of the diagnosis of squamous and glandular intraepithelial lesions in differing age ranges at 5 year intervals, presented a major tendency among the LSIL in younger women (15-25 years of age) and in the cases of HSIL and CA, for the > 50 age group. As to the glandular variations such as adenocarcinoma in situ, this occurred in the > 30 age group and adenocarcinoma in the > 50 age group. Two (0.16%) cases of HSIL were detected in women under 14 years and 51 (4.06%) between 15-19. We concluded that the Pap test is an important tool for detection of cervical cancer especially when associated with internal and external quality control of the Pap smears Our data reinforces the necessity of educational campaigns, investigation and follow-up of the patients for the improvement and effectiveness of the programs of prevention. The human papilloma virus is the main viral agent of sexually transmitted diseases (DTS). In Brazil, according to DATASUS, cancer of the penis is very frequent and there is evidence of the pres-ence of HPV in up to 75% of partners of women suffering from carcinoma of the uterine cervix. The aim of this research was to evaluate the frequency of high and low risk HPV in men. We evaluated 156 men who underwent a first examination without history of previous treatment and showed signs and symptoms compatible with sexually transmitted diseases in any age range. These men were submitted to hybrid capture (CH2) for high risk HPV (HR-HPV) and low risk (LR-HPV) and a histopathological examination when anogenital warts were present. The average age was 38.5 years . Of 156 men, 55 (35.2%) were CH2 positive for HPV and 47 (85.45%) of these were positive for HR-HPV. Of the 49 men with genital warts, 44 (89.80%) presented HPV and 48% positive for HR-HPV. The relationship of penis warts to HR-HPV and LR-HPV was significant: P = 0.0001 and P = 0,006, respectively. Of ten men with anal warts, seven presented HR-HPV and three LP-HPV. Of the six cases identified as HIV positive, 4 (66.67%) presented HR-HPV (P = 0.004). The precise diagnosis of male lesions is fundamental steps in the process of prevention of cervical cancer throughout the world. Our research shows the importance of treating both male and female groups in order to reduce the epidemiological indexes with reference to infection with HPV. P.153 ATYPICAL GLANDULAR CELLS ON PAP SMEAR K. Zachou, E. Tzartza, A. Orologa, A. Skenteri and A. Boni Cytopathology Department of Hippokration Hospital Thessaloniki, Greece

Background: Atypical glandular cells (AGC) on pap smear, is uncommon finding, but may represent a variety of benign and malignant lesions. Objective: The aim of this study is to evaluate the association between AGC on pap smear and histological findings to tailor management protocols. Results: Out of 84.509 Test Papanicolaou which were performed in Hippokratio Hospital of Thessaloniki, between 1999 and 2007, 682 cases (0.81%), were diagnosed as ASCUS, 803 (0.95%) as LGSIL, 550 with HPV infection, 279 (0.33%) as HGSIL, 71 with HPV infection and 46 as AGC (0.05%). Women with AGC on pap smear underwent colposcopy, directed biopsy, endocervical curettage, endometrial sampling and cervical conisation to determine the cytological and histological correlations of AGC on pap smears. The mean age of the patients with AGC was 50, 4 years (rage 32-79 years). Of these patients 22 were postmenopausal and 24 were in reproductive age. We found significant pathological findings including HGSIL, CA in situ, adenocarcinoma of cervix, and endometrial carcinoma in six cases. Conclusion: AGC on Pap smear was associated with a clinically significant diagnosis in approximately 13% of our cases. The women with a diagnosis of AGC on cervicovaginal smear are needed to be evaluated at least with colposcopy, endocervical and endometrial curettage. Clinicians should be careful about the significance of AGC on pap smear. Objectives: It is well known fact that almost all cervical lesions are induced by persistent HPV infection. Incidence of HPV infection among young women is high, but fortunately for them not every infection results with disease. Despite of having good tools for detecting epithelial changes still one can not be sure which lesion will progress, so additional diagnostic or therapeutic efforts must be made. Aim of the study: By performing immnustaining on slides from cervical smears with cellular protein p16INK4a and viral capsid protein HPV L1 we tried to predict which cervical lesion will progress or regress. Materials and Examinees: In our study we followed-up 56 patients with abnormal cervical smears by repeating their Pap smears every 6 months for two years after performing immunoassaying. All of the patients were HPV positive. Slides for immunoassay were taken from the archive or simultaneously, with initial Pap smear and were stained with p16INK4a and HPV L1 according to manufacture proscriptive. After at least two negative Pap smears we could say that initial cervical lesion regressed.

Results: The correlation between L1 and p16 expressions suggests that L1 negative/p16 positive and double positive cases persist or have the potential for progression with one case of regression. All L1positive cases as well as double negative cases showed regression with two cases of persistence. Conclusion: Performing immunoassaying with p16 and L1 can be helpful tool for predicting the behaviour of cervical low grade lesions and to plan the clinical management of these patients. Results: Atypical glandular cells were diagnosed in 370 (0.43%) smears, severe glandular cell atypia/AIS in 59 (0.07%) and adenocarcinoma in 25 (0.03%). Follow up data could not be obtained in six patients with atypical glandular cells. At follow-up examinations 294 (80.7%) patients with atypical glandular cells were negative, 19 (5.2%) had CIN1, 40 (11.0%) had CIN3/CIN2, 3 (0.8%) had squamous carcinoma, 1 (0.3%) had AIS, 5 (1.4%) had adenocarcinoma and 2 (0.5%) had AIS and CIN3/CIN2. Among the patients with severe glandular atypia/AIS 16 (27.1%) proved to be negative at follow-up, 4 (6.8%) had CIN1, 26 (44.1%) had CIN3/CIN2, 1 (1.7%) had squamous carcinoma, 9 (15.5%) had adenocarcinoma and 3 (5.1%) had AIS and CIN3/CIN2. In the patients diagnosed with adenocarcinoma cells in the smear, histology showed adenocarcinomas in 23 cases (92%) and CIN3 or squamous carcinoma in the other two cases (8%). Conclusions: 19% of patients with cytological diagnosis of severe glandular cell atypia/AIS and adenocarcinoma were overdiagnosed, while 14% of patients diagnosed with mildly atypical glandular cells were underdiagnosed.

Materials and Methods: In the Department of Cytopathology at the Institute of Oncology Ljubljana, Slovenia we re-screened 6813 (12.53%) conventional Pap smears out of a total of 54 373 slides screened in 2006, 2007 and 2008 . Random re-screening of the slides (both negative and atypical) was conducted as a component of the educational program for screeners. Results: Of 6813 Pap smears re-screened, 6147 (90.2%) were assessed the same as in the original screening and 666 (9.6%) were reclassified in different categories. There were 221 (3.2%) smears reclassified from negative to reactive, 73 (1.1%) from negative to LGSIL, 8 (0.12%) from LGSIL to HGSIL and two cases (0.3%) were reclassified from negative to HGSIL (true false negatives). Five cases of HGSIL were histologically confirmed CIN2 and CIN3, one CIN 1, in four other cases histology was not yet registered in central registry. Of the 349 cases which were considered to have been overdiagnosed, 219 were reclassified from reactive to negative, 116 from LGSIL to negative, 11 from HGSIL to LGSIL and 3 from HGSIL to negative. Conclusion: In our laboratory we have limited personnel which make rapid re-screening of all negative smears too difficult, subsequently a random re-screen of smears during the screeners' educational program is an adequate policy for quality control. Introduction: Uterine neoplasias require treatments based on surgical resection, radiotherapy and chemotherapy. The realization of cytology is mandatory in the follow-up. The cellular changes associated with radiation are multinucleation, bizarre cells, proportional cytoplasmatic and nuclear increase, anisocariosis and prominent nucleoli. These cellular changes may difficult the differential diagnosis with neoplasic alterations. Objective: Analyze vaginal smears from women treated with radiation. Materials and Methods: Between 2005 and 2008, 124 vaginal monolayer smears were analyzed from 54 women submitted to radiation due to uterine malignant neoplasias. The range of ages varies between 29 and 88 years. The primitive tumours were squamous cell carcinoma (34), endometrial adenocarcinoma (15), endocervical adenocarcinoma (3), adenosquamous carcinoma (1) and leiomyossarcoma (1). All smears were classified using the '2001 Bethesda System'. Results: Out of 47 smears negative for intraepithelial lesion, one revealed benign glandular cells. Of the 72 (58%) smears with reactive cellular changes, 12 (9.7%) were associated with radiation. 5 (4%) smears with epithelial cell abnormalities were observed, 4 (3.2%) with atypical squamous cells of undetermined significance (ASC-US) and 1 (0.8%) with squamous cell carcinoma recurrence. Conclusion: The use of cytology in the follow-up of patients with previous uterine cancer allows the early detection of recurrence/ persistence of tumour. Radiotherapy may induce cellular changes, more or less marked, that need clinical evaluation for exclusion of other pathology and for adequate treatment.

A. R. Rafati* and S. S. Hashemi † *Islamic Azad University, Sarvestan Branch, Iran, Veterinary Medicine School, Shiraz University, Iran Different media have been introduced and experienced for in vitro maturation of oocyte. The aim of this study was to compare three culture media for in vitro maturation (IVM) of bovine occyte. Slaughter house ovaries are the main source of occyte for IVM and IVF studies. Bovine ovaries were isolated from a local slaughter house and transport to laboratory. Oocyte were aspirated from the follicle and washed 4 times in . Total 1820 compact cumulus occyte complexes (COCs) were aspirated from ovaries. Oocytes were randomly cultured in the first medium (control) consists of TCM-199, HCG, follicular fluid (FF) and antibiotic. In the second and third media, different concentrations (10, 15 and 20%) of rat estrus serum (RSS) or estrus sheep serum (ESS) were added to control medium. Then occytes were incubated in 38.5 c, containig 5% co2 and 95% humidity for 24 hours. The maturation of occytes was jugged according to cumulus cell expansion or observation of polar body in randomly orcein stained oocyte. The result of this study showed that maturation rate was significantly higher in second and third group in comparision with control group (P < 0.001), there was no significant difference between second and third groups. Introduction: According to the register for cancer on the theritory of Serbia, every year registrate new 1400 cases of cervical cancer. Incidence is 27.2% at 100 000 of the women. By the occurance frequency the cervical cancer is on the second place, after the breast cancer.

Objectives: The objective of this paper is to present prevention of the cervical cancer in Serbia. Materials and Methods: National program for cervical cancer screening in Serbia is approved in 2008, year by decision of the Government of the Republic of Serbia. At the primary health protection of the women, as a part of the national screening in DZ Savski venac from the 11 500 invited women, only 19.1% of patient came to the medical examine. The methods of the diagnostics are colposcopy and cytodiagnostics. The cytology smears are classified according by Bethesda System and Pap test. Patients are examined by cytologist and supervisor. Women that have first smear normal repeated test after one year, and then every three years. Reports are sending to the gynecology and women received a personalized letter informing them of the test results. Data processing was done using standard statistical procedures. Results: On sample of 2200 patients having first cytology examine 30% of the women have unsatisfactory results, normal results have 63.8%, inflammatory 3%, LSIL 3%, HSIL 0.1%, and invasive carcinom 0.1%. Patients with inflammatory diagnosis are curing and after new examine the test results are normal. Results of histopathology finding show 3. 6% of patients with SIL diagnosis and 3% of patients with ASCUS witch diagnostics by supervisor.

Conclusion: The number of tested patients is small. The great number of tested women is 40 to 50 ages.

V. Balboa*, C. Padilha* † , L. Padilha*, M. Paim* † , R. Agnese* † and R. Carvalho* *Radqualycenter, Rio de Janeiro, Brazil, Cancer National Institute, Rio de Janeiro, Brazil

Objective: Present a new and more effective model in breast and gynecological cancer prevention. Materials and Methods: The project is based on 1 -ACTIVE SEARCH 2 -QUICKNESS in diagnosis and treatment (SAME DAY) 3 -Primary prevention CONSCIENTIZATION.One Mobile Unit of Prevention in Health -MUPH -a truck specially adapted to collect (Pap Test, breast FNAC, breast core biopsy, and cervical biopsy), to do mammography and to emit diagnosis (X-ray, Pap Test, colposcopy, breast cytopathology), is taken to a poor community. The patients are registered, and examinations and diagnosis do ON THE SAME DAY. For gynecological cancer prevention, on those with Pap Test alterations a colposcopy is done, where ambulatory treatable lesions are treated (see and treat method) and, if necessary, medicated for disturbed vaginal flora. In cases of hospitalar treatment the biopsy is done and the patient is sent to the oncologic hospital with the complete written reports, where it also the biopsy result and the slide will be sent. They are previously scheduled after no more than 15 days. Breast Cancer Prevention: Patients indicated do mammography. Those with nodules are punctioned (FNAC) and then if is a solid nodule, the core biopsy is done. Conscientization -Speeches and demonstrations about prevention in uterine and breast cancer, STD and family planning, are ministered in a tent beside the truck, while patients are waiting and also blood pressure and glucose checked. ', 196 women (1. 3%) underwent colposcopic evaluation as a personal request and without having a prior or concurrent history of SIL and had a positive colposcopy and/or a histologically confirmed cervical intraepithelial lesion. Smears of all these 196 women were reviewed by two cytopathologists who searched for the presence of patches of anucleated squames with angulated edges, in conventional and liquid based cytology smears. Results: In 33 (16.8%) out of 196 otherwise negative Pap smears, the additional cytologic criteria of hyperkeratosis were identified (as patches of anucleated squames) which tend to occur in aggregates or overlapping clusters that stain orange or yellow. All these 33 cases were corresponding to biopsy proven low grade SIL. Conclusion: Hyperkeratosis in cytology specimens reported as negative for malignancy (according to 2001 Bethesda system terminology), could be used as a first sign of a possible underlying intraepithelial lesion. For this reason, women with negative Pap smear but with presence of hyperkeratosis, should be referred to colposcopic evaluation. Objective: The aim of this study was to evaluate the effect of estrogen production on the vaginal smears in postmenopausal women, by determining the maturation index (MI). Materials and Methods: Maturation index is expressed as a ratio of the percentages of parabasal, intermediate and superficial cells identified in the smear. Over a period of five years (2003) (2004) (2005) (2006) (2007) , eighty-six (86) healthy women over 50 years of age, at least 1 year after the menopause, had a vaginal smear examined to the Cytology Department of Alexandra Hospital, with a cytological diagnosis 'negative for malignancy' but with an additional indication of high maturation index, as a means of determining an increased estrogen activity, which is not corresponding with the age of the patient. These (86) women were free of gynecological history (rectocele, cystocele, cervical polyp, proptosis uteri and hormonal replacement therapy). The maturation index was also considered high, when the superficial cells were over 20%. Results: In three women(7.5%) with high maturation index, during follow up, were identified histologically proven invasive adenocarcinomas (two adenocarcinomas of the ovary and one endometrial adenocarcinoma). In five women(12.5%) the increased MI was corresponded to benign lesions that usually can cause an effect to the vaginal epithelium (one endometriosis,one leiomyoma of the uterus,one endometrial polyp and two rectoceles). The vaginal smears of the remaining 30 women(75%) became atrophic over the period of the next 3 years. Conclusion: An identification of a high maturation index on the vaginal smears in postmenopausal women, indicates an increased estrogen activity and should lead to a clinical investigation, in order to exclude carcinomas of the genital tract. Introduction: The endometrial tissue is a sensitive target for steroid sex hormones. The histological effect depends on the type of the hormone, the potency, dosage, and the host receptor status. The progesterone therapy produces an arrest of glandular proliferation, with secretory changes in the glands and decidual reaction in the stroma. The endometrial tissue appears quiescent, with no mitotic activity. We present a case of tumour-like decidual reaction of the endometrium in an adolescent, with emphasis on cytologic features of the lesion. Case Report: A 12-year-old Caucasian virgo girl presented with 'vulvar' lesion. A physical examination revealed red elastic mass that prolapsed from her vagina, nearly 6 cm in length without any evidence of vaginal hemorrhagia. She reported progesterone therapy (Premolut), which was prescribed to her for dysfunctional uterine bleeding and had lasted for 20 days. Standard biochemical blood tests were carried out without revealing any pathology or pregnancy. A smear from the lesion was taken and sent for cytological examination and subsequently the lesion was excised and sent for histology. Microscopic examination of the smear showed occasional large eosinophilic cells admixed with abundant small glandular cells and a few squamous cells. Histology revealed decidual reaction of endometrium with aggregation of granulocytes and occasional glandular crowding. Conclusion: Iatrogenic hormonal changes of the endometrium may result to tumourlike lesions. Cytology can exclude malignancy, although the cytologic features may be quite unusual. Excisional biopsy will provide the final diagnosis. COULD FLOW CYTOMETRIC EVALUATION  PLAY A SIGNIFICANT ROLE AS INTERNAL  QUALITY CONTROL OF A CYTOPATHOLOGY  LABORATORY? C. Kottaridi*, J. Georgoulakis*, D. Kassanos † , A. Pappas † , A. Spathis*, N. Margari*, D. Aninos* and P. Karakitsos* *Department of Diagnostic Cytopathology, University General Hospital 'Attikon', Chaidari, Greece, 3rd Department of Obstetrics and Gynecology, University of Athens, Attikon Hospital, Chaidari, Greece

Objective: Cervical cancer is the second most common cancer in women worldwide comprising a major concern of public health. Liquid-based cytology provides significantly more effective detection of low grade intraepithelial neoplasia or more severe lesions, without loss of diagnostic specificity and reduces the number of cases classified as unsatisfactory and false negative cervical smears. The objective of the study is the evaluation of flow cytometry as a rapid tool for quality control of the liquid specimen adequacy for the purpose of precise cytological diagnosis in detecting cervical abnormalities. Materials and Methods: A total of 1854 women undergoing routine cervical cytology joined the present cohort study that took place in Attikon University General Hospital. A monolayer smear was prepared and 1 ml aliquot was removed and prepared for flow cytometric analysis. Cells were fixed and run on a Partec CyFlow SL, with front-scatter (FSS) and side-scatter (SSC) set on logarithmic scale. The forward scatter versus side scatter dot-plot was used for the distinction of ectocervical, endocervical and polymorphonuclear cells. Results: In the total of 1854 cases, 15 specimens (0.8%) were characterized by the cytopathologist as unsatisfactory. The flow cytometric analysis of the same samples confirmed the inadequacy of two out of the 15 samples, as no endocervical cells were distinguished by light scatter while two more samples were found to contain many degraded endocervical cells, as suggested by the reduction of their side-scatter. Monolayer smears were re-prepared for the remaining 11 samples, in order to verify whether slide preparation was the reason for their inadequacy. After the second microscopic observation six out of the 11 cases (54.5%) were found to be satisfactory while the remaining five were still designated as unsatisfactory by the cytopathologist. Conclusion: The parallel screening of adequacy by quantification of cell populations with flow cytometry, is feasible and requires only 1 ml of the LBC sample. It can serve as an internal control for diagnosis and can overall decrease the number of unsatisfactory samples. Objective: The aim of our study is to estimate the prevalence and distribution of oncogenic HPV genotypes in liquid based cytology samples according to Bethesda system cytological results in the same specimen. Materials and Method: Liquid based samples (PreservCyt liquid media) were collected from 1000 women who underwent Thin Prep Pap test and HPV DNA test either for screening or because of a previous abnormal cytology. The HPV typing was done with two methods. The first typing was done with a Multiplex PCR amplification kit. The second method was clinical Arrays HPV which detects up to 35 HPV genotypes. Finally all samples had a PCR screening test (SCR) which permitted detection of presence of HPV DNA through the amplification of a specific fragment of the DNA of the virus using consensus primers. Results: 594 specimens (59.4%) were positive for high risk (HR) genotypes, single or multiple. The concomitant infection with low risk genotypes was circumstanly found.The HPV types identified in order of decreasing prevalence were:

(1) In all the specimen: 31, 16, 53, 66, 58, 33, 18, 51, 52, 59, 68, 56, 39, 70, 26, 85, 73, 35, 45. (2) HGSIL: 16, 31, 18, 51, 53, 56, 58, 59, 73, 84. (3) LGSIL: 16, 31, 18, 51, 52, 56, 33, 58, 53, 59, 68, 70, 39. (4) HPV: 16, 31, 18, 51, 66, 33, 58, 53, 26. (5) ASCUS: 31, 53, 33, 16/66, 58, 51, 52, 59/39, 70/26. (6) SCC: 31, 66 (one case) (7) AdenoCa: 16 (one case) (8) 31, 58, 18, 33, 16, 52, 51, 66, 59, 73 . Conclusion: In our group of women the first in the prevalence of the HPV was the type 31, while HPV 16 was the second. HPV 16 which is the most common type with the biological advantage of transmission, persistency and transformation has the highest prevalence in our study in HPV infection, LGSIL, HGSIL and AdenoCa of the cervix. Introduction: This paper presents the importance of the cytodiagnostics in the early finding of cervical cancer in postmenopausal women. Objective: The objective of this paper is to presents sensitivity of the Pap test in postmenopausal women. Materials and Methods: Due to abnormal cytology and colposcopy finding in 150 postmenopausal women tested on Gynecology and Obstetrics Clinic Narodni front Belgrade, the examination was repeated. Data processing was done using standard statistical procedures. Results: Out of the total number of women examined, PA II group was find in 75% of patients, ASCUS was find in 15%, HSIL was find in 5% and LSIL was find in 5% of patients. The curettage treatment of cervical canal was carried out on patients with ASCUS finding. The colposcopy examination has shown that finding was normal in 70% of patients, in 25% REE was find, REE and punctuation was find in 3.1%, carcinoma was find in 0.6% and HSIL was find in 1.3% of patients examined. Conclusion: Sensitivity of the Pap test is low as well as specificity because the original lesions are not accessible in postmenopausal Objective: The adenocarcinoma of the cervix is a malignant invasion neoplasia expressed by epithelial cells with endocervical origen. The diagnosis of adenocarcinoma has taken on greater significance in recent years, due to both a relative and absolute increase in its incidence. Correctly diagnosing endocervical adenocarcinoma will greatly depend on the grade of the invasive lesion as well as the presence or absence of a tumor diathesis.The authors aim to present a clinical case of adenocarcinoma of the cervix. Materials and Methods: (1) Seventy three year-old postmenopausal women, with metrorrhagias pelvic weight and coital vaginal bleeding.Gynaecologic exam: cervix with marqued vascularisation and intense acidofilic periorificial area.

(2) Endocervical cytology in liquid-based Thin Prep, stained by Papanicolaou. (3) Cervico-vaginal biopsy stained with Haematoxylin -Eosin (HE), Imunohistochemistry (IHC) study (CK7, CK20, CD56, CEA, E12 and VMT) and histochemistry (HC) study with Alcian Blue/PASbP63, CK5/6, 34 with amilase treatment. (4) Radical hysterectomy stained with HE and IHC and HC study. (5) Pelvic linfadanectomy, parametrics and vaginal borders stained with HE. Results: Cervico-vaginal cytology-neoplasic cells, blood background and atipical exocervical cells compatible with HSIL. Biopsy: Low grade poorly differentiated carcinoma of the cervix. The IHC study reveled endometrial or endocervical primitive nature or infiltration by uroterial carcinoma. Histopathologically study of the radical hysterectomy-cervix indiferential adenocarcinoma without uterine body infiltration. This study was complemented with the posterior histological study. Conclusion: The citological diagnostic was extreamely important because guieded the pacient study to the glandular area. This was confirmed by hystology and IHQ study. Introduction: Brazil has one of the highest mortality indexes by cervical cancer around the world. Here the current methodology for this cancer screening is based on conventional Pap test in almost all organized services. NOVOPAP is a liquid-based cytology training center for residents in pathology, a public-private partnership between the Federal University of Sã o Paulo and Qiagen Brasil. Materials and Methods: From April to December 2008, 2413 women from two basic health service unities were evaluated for cervical cancer at NOVOPAP. The Bethesda System was performed and women were sub-classified in two age groups. Data was analyzed in BioEstat 5.0 by qui-square test. Results: There were 262 women aged under 25 (10.9%) and 2151 (89.1%) aged 25 or more. We found 2174 (90%) Negative results, 35 ASC-US (1.5%), 36 ASC-H (1.5%), 70 LSIL (2.9%), 13 HSIL (0.5%), 6 glandular atypia (0.3%) and 74 Non-satisfactory samples (3.1%) . In both age groups, LSIL was the commonest diagnosed atypia. HSIL and glandular atypia were found only in the oldest group. The data showed statistic significance (P = 0.04). Hybrid capture will be performed for all ASC-US, Negative and Non-satisfactory samples as a quality control method monitoring cytological diagnostic since we don't have any follow-up visit notice from these women, so more data is going to be applied. Conclusion: Our experience on liquid-based cytology seams to be like that found in many population-based studies abroad. Objective: The basis for a good quality cervical smear is a correct sampling method. We evaluated a cone-shaped polyurethane sampling device, the Papcone (Otto Bock, Duderstadt, Germany); designed to obtain simultaneously cells from the ecto-endocervix. We analysed its ultrastructure by means of the scanning electronic microscope (SEM), comparing it with the traditional wooden Ayre's spatula (S) and cytobrush (C), and the related smears. Materials and Methods: A 28-years old, primiparous woman was submitted to traditional Pap smear (by Ayre spatula and citobrush), and after 3 months to Papcone sampling. Slides were prepared according to Papanicolaou method and examined by light microscopy. Papcone, S and C ultrastructure were examined by SEM before and after sampling. Specimens for SEM were fixed in 2.5% glutaraldehyde in 0.1M PBS, and stored at 4°C. Samples were postfixed in 1% osmium tetroxide, and then dehydrated in increasing concentrations of ethanol. Specimens were critical-point dried with carbon dioxide, mounted on aluminum stubs, coated with platinum, and observed with a Hitachi S-4000 FE-SEM. Results: Papcone showed lower bleeding and less overlapping cell layers in comparison with S and C. Traditional Pap smears showed a clear separation between ecto-and endocervical cells. SEM evidenced how the Papcone surface appeared porous, partly occluded by membranes, and divided by trabecles lodging sampled cells. S showed a finely regular surface, sometimes showing chips, cells appearing clustered in small groups. C most often entrapped cells at or among the bristles, wherein red cells were usually noted. Conclusion: Papcone is a good method to obtain cervical samples and it can be useful especially in case of flogosis or cervical bleeding. Background: Conventional cervical smears may provide important data not only about exocervical and endocervical pathology, but also of endometrial pathology, limited by the involvement of the lining columnar epithelium of the cervical canal and of endometrial cells desquamation.

Objective: The aim of this study was to perform a retrospective correlation of the atypical glandular cells (AGC) observed on conventional cervical smears, stained with Papanicolau technique, with subsequent histopathological diagnosis obtained on biopsies. Materials and Methods: We retrieved 87 cases of conventional cervical smears from our files, with AGC suspicions, 32 of them being considered more likely as reactive changes. All cases were correlated with their cervical biopsies, endocervical and endometrial curettages, paraffin-embedded and routinely stained, during a 3 years follow-up period. Results: Biopsy findings of cervical smears were as follows: 62.06% miscellaneous benign lesions (32 cases, already excluded, were represented by endometritis, endocervical, endometrial, and mixed polyps, irregular proliferative endometrium, adenomyosis, simple and complex endometrial hyperplasia, tubal and squamous metaplasia, and cervical endometriosis and 12 cases were represented by chronic cervicitis), 22.98% malignancies: cervical adenocarcinoma in situ (n = 6), cervical adenocarcinoma (n = 10), and cervical adenosquamous carcinomas (n = 4), and 14.94% squamous intraepithelial lesion: LGSIL (n = 8) and HG-SIL (n = 5). Conclusion: Biopsy follow-up of conventional cervical smears with diagnose suspicion of atypical glandular cells (AGC) revealed mainly benign reactive epithelial changes, followed by in situ and invasive malignant glandular lesions, and squamous cervical intraepithelial lesions. A careful examination and follow-up of glandular lesions is necessary, as in our small series, 37.93% of investigated cases were malignant type and precursors lesions. Objectives: Women with repeat ASC-US or LSIL were referred to colposcopy. At colposcopy samples were taken for HPV testing, cytology and histology. We wanted to compare high risk HPV DNA detection by two different methods. Cytology as well as histology from colposcopies were available for comparison. Materials and Methods: Cytology was assessed by LBC (Thin-Prep). High risk HPV DNA was tested by Hybrid Capture 2 (HC2) liquid hybridization method and Roche HPV Amplicor from the same specimens. Histological sections were stained with HE. Immunostaining for p16INK was done for histological specimens. Results: Altogether 345 patients were examined. When comparing the two hrHPV DNA tests, concordant results were obtained in 281/345 (81.4%) of cases. Of these 55% were HPV positive. Overall, Roche Amplicor test had higher sensitivity and was positive in 57.9%, as compared to 47.5% positivity of HC2. Among the discordant HPV results, histology was available in 61/67 cases. HC2 negative, but Amplicor positive cases (n = 49) included 15 CIN1, 4 CIN2 and one CIN3. HC2 positive but Amplicor negative cases (n = 12) included one CIN1 and one CIN3.

The interpretation of atypical squamous cells of undetermined significance (ASC-US) and cannot exclude highgrade squamous intraepithelial lesion (ASC-H) is very controversial due to standardization of the cytological criteria by interobserver and interlaboratory. Objective: To determine the interobserver variability in the interpretation of ASC-US and ASC-H in cervical cytology specimens. Materials and Methods: Over 879 conventional cervical cytology specimens stained by Papanicolaou method, analyzed by Bethesda System 2001 criteria as ASC-US and ASC-H were revised by two observers. Results: From 640 cases previously interpretated as ASC-US, 433 (67%) were concurrence by the first observer and 410 (64%) by the second one and from 239 cases of ASC-H, 72 (30%) were concurrence by the first observer and 76 (31%) cases by the second one. Using the Kappa test to compare the interpretation among the observers the concurrence was minimal (< 0.40) to the confidence range of 95% and P < 0.001 and grade by the general Kappa index (0.248). Conclusion: There was a great variation in the interpretation of the two interobservers compared to the previous analyses, but between the two ones there was a good one concurrence. Several factors could be contributed to the fair interobserver reproducibility of adequacy assessment as obscured smears with great number of red and white blood cells and air drying artifact. This indicated the needed to provide a more realistic gauge of interpretive reproducibility on the diagnosis criteria standardization and particularly in the interpretation of ASC cases, several observers could be required. Introduction: Genital HPV infections are one of the most common sexually transmitted diseases caused by human papillomaviruses (HPV). These viruses are thought to be the main etiological factor in cervical cancer development. Greatest percentage of the determined cases of carcinoma are located on the cervix uteri. This percentage could greatly be decreased by an efficient functioning of prevention in medical care of women. Aim of the study: The aim of this study is to evaluate the extent to which colposcopy, cytology and targeted biopsy may be used in HPV diagnosis so that treatment is promptly administered and possible malignity development prevented. Materials and Methods: Retrospective cohort analysis was used to test 1148 women that underwent colposcopy, cytology (Pap smear) and targeted biopsy testing during the 2008 at the Clinic of Gynaecology and Obstetrics 'Narodni Front', Belgrade. Results: Changes in the cervix were found in 1.000 out of 1148 tested women; 56 in the vagina, and 92 in vulvae. HPV infection was found in 484 women where cervical changes were detected; for the remaining 516 women the infection was not diagnosed. Colposcopy testing showed thet abnormal results more frequently occured in HPV-positive subjects (v 2 = 17.97; P < 0.05). Cytology results showed that HPV infected women usually exhibited group III of the Pap smear test, but HPV-negative subjests exhibited group II (v 2 = 603.37; P < 0.05). A positive pathohistology test more frequently occured in HPV-positive subjects, whereas HPV-negative subjects predomintly had normal test results (v 2 = 351.94; P < 0.05).Women with HPV infection demostrate a low grade dysplasia (v 2 = 200.4; P <0.05).

Conclusion: All three methods should be used in conjunction to diagnose HPV infection. Regular screening and detection of premalignant changes will reduce the incidence of cervical cancer in women. Objective: In this work, we evaluate the utility of computer monitoring of HC2 positivity in order to identify possible deviations in the cytological interpretation. Materials and Methods: In the software SGAP (used in our lab) is registered the cytological interpretation proposal (preliminary report), the pathologist interpretation (definitive report) and the HPV test result by hybrid capture 2. The SGAP software permits, for a determined period of time, to obtain the positivity percentages for each interpretation (Nilm, ASC, LSIL, HSIL, etc) in the preliminary or in the definitive report. The software permits also, the identification of cases for review. Were analyzed data from January 2007 to April 2008. Results: The SGAP refered to an excess of HC2 positivity from one of the cyto-technician in NILM category. The NILM HC2 positive cases were revaluated by the same cyto-technician. The result of this action was analyzed between April 2008 and December 2008. Objective: The ThinPrep Ò system is a liquid based cytology which its vial (containing preservcyt medium) is processed in a automated device. Currently there are three models available (TP2000, TP3000 and the new TP5000). With the introduction of molecular biology techniques, the value of liquid cytology samples increased due to the possibility of performing these techniques in the same vial. However, this is conditioned to the remaining material when the cytological preparation is primary done. This work intends to evaluate the impact in the remaining volume of preservcyt after the cytological processing when using the new TP5000 comparing with the TP2000. In fact, the most wide spread HPV test, hybrid capture 2, requires at least 4 ml of preservcyt remaining after cytological processing (in HPV reflex testing). For this work we consider that below this value we have insufficient residual volume. Materials and Methods: 6303 samples were processed with the TP2000 in the selected period and 3011 with the TP5000 until the present date. Results: 3011 cases were processed with the ThinPrep 5000 Ò and 6303 with the ThinPrep 2000 Ò . The number of cases which the remaining volume after processing was less than 4 ml is 169 for the ThinPrep 5000 Ò (5.61%) and 264 for the ThinPrep 2000 Ò (4.19% Objective: The objective of the study was to analyze the follow-up tissue diagnoses after a cytological interpretation of 'low grade squamous intraepithelial lesion, a more severe lesion can not be ruled out' (LGSIL-more). ' LGSIL-more' interpretation includes lesions that have characteristics of LGSIL lesions with additional features that do not completely satisfy a HGSIL interpretation. A further comparison was made between tissue follow-up of the cytological interpretation of 'low grade squamous intraepithelial lesion' (LGSIL) and LGSIL-more. Methods: A review of patient records from years 2005-2008 revealed that 147 cases had cytological findings of LGSIL-more and 415 cases had cytological findings of LGSIL. Surgical tissue follow up study results were retrieved and correlated with the findings on cytology. Results: Follow up of patients with the diagnosis of LGSIL-more revealed that 11% of these patients had benign histological findings, 57% grade 1 cervical intraepithelial lesion (CIN I), and 32% of these patients had grade 2 cervical intraepithelial lesion (CIN II) or higher. In comparison, 23% of patients with LGSIL had benign histological findings, 70% had CIN I and 7% had CIN II or higher.

The majority of patients with LGSIL-more cytological diagnosis had CIN I on surgical follow-up studies. However, a greater percentage of patients with a LGSIL-more cytological interpretation had a CIN II or higher lesion on tissue follow-up as compared to the patients with LGSIL interpretation. This suggests that a cytological interpretation of LGSIL-more may warrant closer clinical surveillance. Objective: The objective of this study was to evaluate the frequency and the significance of cytomorphological criteria defined in previous studies as being predictive of neoplasia in cervical smears of women with a cytological diagnosis of atypical glandular cells (AGC) or adenocarcinoma in situ (AIS). Materials and Methods: Women (n = 103) with cytological findings suggestive of AGC or AIS, whose diagnoses were later established by histopathology, were included in the study. The criteria analyzed in cervical smears previously classified as AGC-NOS (not otherwise specified), AGC-FN (favor neoplasia) or AIS were: irregular nuclear membranes; scanty cytoplasm; dyskeratotic cells; increased nuclear/cytoplasmic ratio; the presence of nucleoli; overlapping; papillary clusters, feathering; loss of polarity; nuclear enlargement; coarsely granular chromatin; and pseudostratified strips. For statistical purposes, odds ratio (OR) with 95% confidence interval (95% CI) were calculated. Results: Histopathology resulted in squamous or glandular neoplastic diagnoses in 55 cases (53.3%). Coarsely granular chromatin was observed in 62.5% of cases with a diagnosis of neoplasia. Feathering was present in 80% of cases of AIS confirmed by histopathology. Loss of polarity and coarsely granular chromatin were significantly associated with neoplastic diagnosis showing OR values with OR values respectively of 6.12 (95%CI 1.13-43.60) and 3.50 (95%CI 1.13-9.44). In AGC-SOE subclassification, coarsely granular chromatin was significantly associated with neoplastic diagnosis (OR 95%CI 4.50 1. 02-22.72 Introduction: The most important event in cervical cancer carcinogenesis is the infection by HPV and the integration of the viral episome in cellular genome. However, it is still unknown the 'timing' of progression between the developement of the lesion in squamous cervical cancer, after viral integration. P16INK4A is a cyclin-dipendent kinase inhibitor, negatively controlled by RB gene product. The protein is tightly correlated to viral DNA integration and with the expression of two viral oncoproteins: E6 and E7. Methods: 137 women, who underwent to Pap Test using Liquid Based Cytology (LBC) technique, were observed in the period 2006-2008. In 78 cases the corresponding bioptic sample was also available. We observe the distribution of the lesions, the cyto-histological correlation, the presence of HPV as detected HC2, the incidence of p16 (CIN Tec) in cyto-histological samples. Results: The incidence of no negative Pap-test has been of 310/12312 (2.5%). In none of the 48 evaluated cases with Negative cytology, including 6 HPV+, p16 immunoreactivity (IR) has been evidenced. IR has been found in 1/22 cases ASCUS/AGUS, in 4/20 L-SIL, in 6/13 H-SIL and in 2/4 SCC. In 51/78 cases, of which the histological sample was available, the p16 IR has been determined: a strong IR was present in 1/28 negative cases, 2/8 lesions CIN1, 9/12 CIN2-3, 3/3 SCC and in 2 lesions condylomatous. In the evaluable cases (excluding ASCUS, AGUS and L-SIL) the Overall Agreement was 91% (83.3% with LSIL). 2/14 cases cytologically classified as L-SIL were confirmed by histology, while on eight cases histological lesion CIN2+ was found and in 4 last cases any lesion was evidenced. In 18 cases, p16 has been determined both on the cytological and histological samples: in two was present IR only on the cytological sample, while in one case the opposite event was observed. In 75 cases the presence of HPV has been determined: 15 case p16 IR positive also presented infection by Low and High Risk-HPV (LR-HPV and HR-HPV); in 21/ 44 cases that didn't express p16 IR, LR-HPV and HR-HPV infection was detected. Conclusions: Our study shows that the presence of p16INK4A protein in cyto-histological samples could be useful in giving information on viral integration also in absence of morphologically detectable lesions.

study to determine the overall HPV prevalence and identify the most prevalent HPV genotypes associated with the lesions. Materials and Methods: Liquid-based cytology samples (Thin-Prep) were collected consecutively for cytological screening. HPV testing was performed in all cases with Hybrid Capture 2 (HC2; Digene) (high-risk probes), and positive or inconclusive tests were genotyped using PapilloCheck Ò test (GreinerBio-One). We analysed samples from 587 women, classified according to the Bethesda System: 62.0% NILM, 9.0% ASC-US, 21.0% LSIL, 5.0% HSIL, 1% squamous cell carcinoma, 1% atypical glandular cells (AGC), 0.5% adenocarcinoma and 0.5% others. Results: HPV was detected in 351/587 (60%) cases and 126/587 (36%) were multiple infections. HPV 16 was the most prevalent genotype (21% of all women), and was also the most prevalent in women with HSIL (49%) or with squamous cell carcinoma (67%). Other common HPV genotypes in women with HSIL included HPV 18, 31, 52. HPV 16 and/or HPV 18 alone were present in 61% of HSIL and in 77% of cervical cancers. Conclusion: This suggests that an efficacious HPV16/18 vaccine will have a substantial preventive potential in the general female population. Cervical cancer is one of the most important causes of death, particularly in undeveloped countries, including Uruguay, representing a major public health problem. Although in our country a screening program of cervical cancer has been implemented in the last few years, no study about the prevalence of cervical squamous intraepithelial lesions (SIL) has ever been reported. The aim of this study was to determine the prevalence of SIL in a uruguayan women population and to investigate the distribution of HPV genotypes. Cytological samples were processed for conventional cytology applying Bethesda system, 2001. In case of abnormal cytology, colposcopy and eventually biopsy were conducted. HPV DNA was investigated in 287 of these samples; 167 Low grade SIL (L-SIL), 37 High grade SIL (H-SIL), an adenocarcinoma and 82 control samples. HPV DNA was detected by PCR using MY09/MY11and/or Gp5+/Gp6+ consensus primers and genotyped by RFLP and/or sequencing. Overall, HPV DNA was detected in 24.7% of women. In L-SIL, H-SIL, and Adenocarcinoma samples, the rates observed were 25% (42/167), 67% (25/37) and 100% (1/1), respectively. Similarly, HPV DNA was detected in 12 (14.5%) out of 82 control women. In all cases, HPV 16 was the genotype most commonly found. This report represents the first study carried out in Uruguay about SIL prevalence and HPV distribution. Results notably the low HPV prevalence rate in SIL, particularly in those categorized as low grade. In agreement with almost all studies reported elsewhere, we found out that also in Uruguay HPV 16 is the most widely distributed genotype. Genotype HPV 18 was no detected in any case. The results reported in this study might represent useful information in determining the impact of HPV vaccine program implementation. Objective: The role of HPV in cervical cancer carcinogenesis is nowadays well established, as is the regression capability of its precursor lesions. Several authors have proposed that HPV L1 and p16INK4a together can be helpful for estimating the biologic potential of precursor lesions. We tested the use of Cytoactiv Ò HPV L1 Screening Set, on ThinPrep Ò PAP stained slides, in which expression of p16INK4a had been previously detected. Materials and Methods: Ninety seven ThinPrep Ò PAP stained slides, in which p16INK4a immunocytochemical staining had been previously performed, were retrieved from our files (years 2000 to 2005) . In all cases HR-HPV DNA had been assessed by HC2 Ò (reflex test). After cover slip removing, Cytoactiv Ò HPV L1 Screening Set (Cytoimmun, Germany) was performed according to manufacturer's instructions (antigen demasking: water bath at 95°C for 20 minutes). Positive and negative controls were used in each round. A positive threshold of one stained nuclei was used. Results: Cell morphology and preservation remained intact allowing easy identification of cell type, with the positive red nucleus. Double stained red/ brownish cases were also easy to recognize. Seventy two cases were HR-HPV DNA positive, and were classified on the following cytological categories: ASC-US(34), LSIL(19) and HSIL(19) . Negative HR-HPV DNA cases were 25: NILM(17) and ASC-US (8). There was no staining of HPV L1 in this group, but p16INK4a was positive in 13 cases (7 NILM and 6 ASC-US). In the HR-HPV positive group, HPV L1 was detected in 24 cases (33.3%): 9 ASC-US, 10 LSIL and 5 HSIL; p16INK4a was positive in 41(56.9%) cases: 17ASC-US, 7 LSIL and 17 HSIL. Thirteen cases (18.0%) were HPV L1+/p16INK4a+. Twenty cases (27.8%) were HPV L1-/p16INK4a-. Conclusions: Cytoactiv Ò HPV L1 immunostaining method is a simple, easy to read method, without subjective interpretation. Double staining with Cytoactiv Ò and p16INK4a is also easy to interpret and LBC proved to be versatile and resistant, since cell morphology was maintained after two different rounds of antigen retrieval technique. This double staining may prove to be useful for retrospective and follow-up studies. Objective: One of the advantages of liquid-based cytology over conventional Papanicolaou smears is the significant reduction of unsatisfactory samples. ThinPrep Ò manufacturers (HologicTM) recommend a reprocessing protocol for gynecologic cytology samples, considered as poorly cellular at the first screening, in order to further diminishing the number of unsatisfactory cases. We reviewed a series of consecutive cases in which this protocol was applied in order to evaluate its impact in reducing the number of unsatisfactory samples. Materials and Methods: We reviewed 221 cases, out of 4000, in a 9 month period, considered by cytotechnicians, at first screening, as unsatisfactory due to low cellularity or obscuring blood, which were reprocessed according to the HologicTM protocol. The protocol consists on removing a 5 ml sample from the vial, centrifugating it and adding to the pellet 30 ml of acetic acid/CytoLyt (glacial acetic acid in CytoLyt 10%). The sample is then centrifugated again and the resulting pellet is added to a new vial of PreservCyt and processed in the TP2000. The use of just 5 ml of the initial sample will still allow search for HPV-DNA in the remaining sample, if needed. We consulted the final cytology result of these cases, considering specimen adequacy and number and type of abnormalities found. Results: From the 221 samples reprocessed, 186 (84%) were classified as satisfactory in the cytological report and 35 (16%) remained as unsatisfactory. From the 186 cases successfully reprocessed, 28 (15%) were considered as abnormal, being 18 cases classified as abnormal squamous cells (11 ASC, 3 LSIL, 4 HSIL) and ten abnormal glandular cells (9 AGC, 1 ADC). Follow-up of these cases confirmed 4 CIN III, 3 CIN I and 2 ADC. Conclusion: The protocol for reprocessing samples recommended by HologicTM reduced the number of unsatisfactory samples in 84% and identified 28 (15%) cases of abnormalities of epithelial cells, which otherwise would not have been diagnosed. Objective: Infection with oncogenic HPV types is acknowledged as the major risk factor for the development of cervical cancer. However, the vast majority of the infections will spontaneously regress or disappear while a small a percentage will persist or progress to cervical intraepithelial neoplasia. The pattern of E6-and E7 mRNAs expression is concerned a functional discriminator between high-and low-risk HPV lesions. Recently, a new technique for the detection of HPV E6-and E7-mRNAs using the high-throughput capabilities of flow cytometry has been described. In this study we investigated the significance of HPV E6-and E7 mRNAs in women with cervical intraepithelial neoplasia. Materials and Methods: Our study included 150 cervical smears obtained by 140 women. The specimens were processed by Thin-Prep method and were diagnosed according to the BETHESDA SYSTEM. In the rest Preserv material of each case, flow cytometry (HPV OncoTect Invirion Diagnostics LLC) and Hybrid Capture 2 were performed. Twelve cases underwent follow-up after 3 or 6 months. The results of these methods were evaluated in correlation with Cytomorphology. Results: This assay was performed in less than 3 hours directly from Thin Prep specimens. All control cases were negative. The percentage of E6-and E7-mRNA expressing cells in HSIL was greater than in samples diagnosed as LSIL or ASCUS. In a small percentage of specimens diagnosed as WNL flow cytometry assay was positive at the first control and negative in follow-up. Conclusions: E6-E7-mRNA testing is a rapid, easy method and seems to be more appropriate for risk evaluation, especially in younger age groups. This method is more concordant with cytomorphology than HPV DNA test. Persistent expression of E6/E7 oncogenes could serve as an indicator of progression to cervical intraepithelial neoplasia and invasive cancer. Our study suggests that the detection of mRNAs E6/E7 using flow cytometry, in comparison with other assays relied on viral DNA, could lead to a more appropriate management of cervical intraepithelial lesions. Further prospective studies are necessary in order to confirm these findings. This study aimed to verify one of the major diagnostic dilemmas in routine Pap smears regarding ASCUS and AGUS was tested immunocytochemically for p16INK4A (p16). The results were correlated with follow-up biopsies. The study designed included 66 Pap smears of ASCUS diagnostic categories and 36 Pap smears of AGUS diagnostic categories, reclassified according to the 2001 Bethesda System, all of which were correlated histologically. Pap smears were decolorized and immunostained with the primary anti-p16 antibody -clone E6H4. Increased levels of p16 were related to infection of high-risk human papillomavirus (HR-HPV), most possibly via the production of the viral oncoproteins E7. Increased expression of these HR-HPV oncogenes in abnormal cervical cells was reflected in an over-expression of p16. Conversely, lesions mimicking those cervical neoplasias discussed above and showing no evidence of HPV infection will not exhibit elevated expression of p16. The results revealed that of the 66 smears containing ASCUS, 47 (71%) was reclassified as ASC-US and 19 (29%) as ASC-H. Follow-up biopsies revealed that 21 (32%) cervices had no obvious abnormalities but only reactive changes. A significant proportion of histological diagnoses were CIN1/LSIL (24 cases, 36%), CIN2 or 3/HSIL (17 cases, 26%), squamous cell carcinoma (two cases, 3%), or AIS/adenocarcinoma (two cases, 3%). The p16 immunocytochemical stain was reactive in 40 (60.6%) of 66 smears: either weakly/sporadically (18 cases, 45%), or strongly positive (22 cases, 55%). Conversely, 26 (39.4%) of the smears were negative for p16 and displayed predominantly reactive changes. However, five cases of LSIL and one of HSIL were negative for p16. Of the 36 smears containing AGC, 22 (61%) were reclassified as general AGC and 14 (39%) as AGCfavor neoplasia. Follow-up biopsies revealed that 15 (42%) cervices had no obvious abnormalities but only reactive changes. Significant proportions of histological diagnoses were LSIL/CIN1 (2 cases, 6%), HSIL/CIN2 or 3 (12 cases, 33%), adenocarcinoma in situ (4 cases, 11%), and carcinoma (3 cases, 8%). More than half cases (19/36, 53%) of follow-up biopsies concerning AGC-containing smears represent significant lesions. There was much higher proportion of significant lesions (13/14, 93%) found in AGC-favor neoplasia than (6/22, 27%) in general AGC cases. 15 of 36 (36%) AGC-containing cases were actually squamous abnormalities in follow-up biopsies. The p16 immunocytochemical stain was reactive in 22 (61%) of 36 smears: either weakly/sporadically (2 cases, 6%), or strongly positive (20 cases, 56%). Conversely, 14 (39%) of the smears were negative for p16 and displayed predominantly reactive changes. However, there was one case of HSIL showing negative immunostaining for p16. From the viewpoint of clinical significance, both ASCUS and AGUS analyses were highly sensitive and specific, in addition to favorable positive and negative predictive values. In conclusion, on the basis of both morphological and immunostaining patterns, there is a clear association between strong p16 immunostaining of atypical cells in the smears and the presence of significant lesions in the cervix. Similarly, there is a clear association between the lack of p16 expression and the absence of cervical lesions. The p16 immunocytochemical stain can be applied successfully to conventional Pap smears and may serve as a useful biomarker in ASCUS-and AGUS-containing smear diagnoses. This may offer a more objective parameter to help clarify this ambiguous area in gynecological cytopathology. Introduction: Cervical cancer (CC) is the second most common cancer among women worldwide. The German Federal Ministry of Health and Social Security recommends CC screening for all women at the age of 20 years on basis of the Papanicolaou (PAP) smear. HPV testing and liquid based cytology (LBC) should not be integrated. A lot of epidemiological studies have demonstrated that HPV infections are a major risk factor for the development of invasive cervical cancer (ICC). At least 14 of the high risk (HR) subtypes are accepted to be oncogenic (F. X. Bosch, 2008) . German Society of Gynecology and Obstetrics recommends integrating HPV testing into primary screening for women older than 30 years (AWMF 015/027). Most of the female population will get infected with HPV during their lifetime, but generally the infection is eliminated by the t-cell system. Persistent infections with certain HR HPV types (HPV 16, 18 and 45) seem to demonstrate a higher risk to develop CC (Bulk S, 2006) . Materials and Methods: At enrollment two HPV DNA tests were performed. The FDA approved Hybrid Ò Capture II test and the 16/18/45 Probe Set test (HC2T, PST; Qiagen, Hilden, Germany). A total of 586 women were recruited [109 women attending for routine cc screening/477 women with well-known cervical lesion and infections (eligibility criteria: hr+)]. 31 were excluded due to history of conization or hysterectomy (25), pregnancy (2), age (1), hpv negativity (1) and lost-to follow up (3) . Results: The overall baseline prevalence for HPV 16, 18, 45 infections in the risk group is 65.47%. 13.76% of the control group were HC2T positive, respectively 8.25% were PST positive. Discussion: Starting CC screening at the age of 30 seems to be difficult as 17 of the 68 histological verified severe cervical lesions arise in women < 30 years. 86.36% of HSIL were PST positive and progressed to CIS. Results: In decreasing order, the following genotypes were detected: 16 (32.58%), 31 (12.96%), 53 (11.14%), 66 (7.23%), 58 (6.60%), 52 (6.57%), 6 (6.02%), 18 (5.75%), 51 (5.63%), 33 (5.56%), 56 (4.98%), 42 (4.88%), 39 (4.86%), 68 (4.06%), CP8304 (3.92%), 45 (3.05%), 59 (2.49%), 35 (1.91%), 11 (1.89%), 44 (1.43%), 43 (0.48%) and indefinite (10.25%). Conclusions: Several meta-analysis have shown geographic variation in the prevalence and genotype distribution of HPV [3] [4] [5] . In Extremadura, the most common type is HPV-16, and together with HPV-31 and HPV-53 involves a half of infections in our population. Introduction: The prevalence of cervical infection with human papillomavirus (HPV) in women is closely related to the corresponding risk of cervical cancer [1, 2] . Data on HPV prevalence and type distribution are essential to predict the potential impact of prophylactic vaccines against HPV6/11/16/18. Materials and Methods: 600 cervical pap tests negative for intraepithelial lesions or malignancy within five broad age-groups: 24 years and less, 25-34 years, 35-44 years, 45-54 years and more than 54 years were analyzed the presence of DNA-HPV by consensus PCR. Results: HPV DNA was detected in a 6% (36) of cervical pap smears analyzed. HPV prevalence decrease with patience age: 12.50% (24 years and less), 10% (25-34 añ os), 5% (35-44 añ os), 2.5% (45-54 añ os) and 0% (more than 54 years). Conclusions: HPV DNA prevalence in women without cytological abnormalities varies greatly worldwide [3] [4] . In Extremadura, 6% of women with normal cytology are carriers of HPV DNA, with marked variation between age-groups. Alentejo: A total of 290 gynaecological cytologies positive for HPV (detected by PCR) classified as: negative for intraepithelial lesions or malignancy (8), inflammation (7), ASC-US (116), ASC-H (28), LSIL (87), HSIL (38), squamous cell carcinoma (2) and AGC (4) . Results: Extremadura: In descending order the following genotypes were identified: 16 (32.58%), 31 (12.96%), 53 (11.14%), 66 (7.23%), 58 (6.60%), 52 (6.57%), 6 (6.02%), 18 (5.75%), 51 (5.63%), 33 (5.56%), 56 (4.98%), 42 (4, 88%), 39 (4.86%), 68 (4.06%), CP8304 (3.92%), 45 (3.05%), 59 (2.49%), 35 (1.91%), 11 (1.89%), 44 (1.43%), 43 (0.48%) and indeterminate (10.25%). Alentejo: In descending order the following genotypes were identified: 16 (28.97%); 51 (15.17%); 31 (12.41%); 53 e 66 (11.38%); 33 (8.97); 58 (7.93%); 52 e 70 (5.17%); 84 (4.83%); 18 e 61 (4.48%); 82 (4.14%); 59 (3.79%); 6 e 62 (3.10%); 39 e 56 (2.76%); 11, 68 e 83 (2.07%); 42 e 81 (1.72%); 35 (1.03%); 72 e 73 (0.69%); 26, 44, 45, 85 (0.34%). Conclusions: Prevalence and genotypic distribution of HPV varies in different geographical regions worldwide [3] [4] [5] . In Extremadura and Alentejo the most common type is HPV-16. In Extremadura the HPV-16, HPV-31 and HPV-53 are present in more than half of HPV infections in the population, while in Alentejo are the types HPV-51, HPV-31, HPV-53 and HPV-66. Introduction: Nowadays the integral health of women is a major objective for our society to reach the effective equality between men and women. The existence of preventive ways that have already demonstrated their effectiveness in different population groups and the difficulty to access to it prompted the beginning of the 'Cervical Cancer Screening Programme'. The performance of cytology is a preventive way, accessible, simple and beneficial to all women, which will prove his state of health and in case of any anomaly can effectively benefit from other measures that could improve its state. The correlation between cytology and histology is one of the tasks involved in the daily practice of a screening and a tool for cytological diagnosis in acute, with the improvement of the same. Materials and Methods: A total of 107 cytological specimens positive for HPV, classified as ASC-US (28), ASC-H (10), LSIL (29), HSIL (27), AGC (5) squamous cell carcinoma (6) and adenocarcinoma (2) were correlated with their biopsy or surgical specimen. [3] [4] [5] . In our community the most common type is HPV-16, which with HPV-51, HPV-31, HPV-53 and HPV-66 is present in more than half of HPV infections in the population. The aim was to investigate the prevalence of HPV L1 capsid proteins in HPV infected HSIL and LSIL. HPV L1 capsid proteins are considered to be a major target of cellular immune response in cervical intraepithelial squamous lesions. Materials and Methods: Cervical conventional smears from 82 women with cytologically and histologically confirmed LSIL (n = 54) and HSIL (n = 28) were collected retrospectively to detect HPV L1capsid proteins by immunocytochemistry using the monoclonal antibodies (High Risk Antibody VAHP) in a standardized protocol. HPV infection was detected by the presence of cytopatic HPV effect (koilocytes) in the smears and biopsies.

Results: HPV L1-capsid proteins were detected in HSIL and LSIL for HPV high risk in 28% and 40% of the specimens, respectively. Expression of L1-capsid proteins was significantly reduced for HPV positive HSIL. In HPV positive LSIL, no significant reduction of L1 capsid protein expression could be demonstrated. Conclusions: These data support the statement that failure to detect L1 capsid protein in smears and biopsies correlates with progression of the lesion. Because L1 is the major target of the immune response, its loss at early stages of the transformation process may lead to inefficient stimulation of the immune response and promote further transformation of immature epithelial cells. Immunocytochemical detection of HPV L1 capsid protein has prognostic value for the follow up of early displastic cervical lesions. we have a discrepancy in the diagnosis. We didn't have any false positive cases. We had six false negative cases (three by sampling error and three by interpretation error). We found a discordance in grade in five cases. The frequency of discrepancy was 9.4%. Conclusion: Our study emphasize the importance of having a computorized QC data base. Our frequency of discrepancy was within the literature results. The study evaluates the quality of the sample and the overall interpretative skills of the cytopathologist. In the future we will address the question -should the histological result be the only golden standard? Objective: The kinetics of the dyeing procedure of the Adolfo Lutz Institute Modified Papanicolaou (ALIMP) method is analyzed to quantify the minimum value for absorbency and of the quantity of slides that can be dyed with the same solution. Our purpose is the development of the quality control of the ALIMP method.

The ALIMP is a progressive method and is used in the routine work of Adolfo Lutz Institute. The procedure of dyeing was accompanied along 33 working days and the control was done using slides of a woman's vaginal smear. The slides collected in ten different days were analyzed using the program SPCIM (System of Processing Microscopic Images). The slides were classified by eleven cytologists according with ten quality indexes. Results and Conclusions: Slides and analysis that gave discrepant results were eliminated using statistical techniques and paraconsistent logic. Non-parametric statistical technique was used to obtain an average classification of the slides according to indices given by the cytologists. It was observed a good adhesion between the average classification and the average intensity of absorbency. The present analysis gives us the support for the development of a system for the quality control of the ALIMP method using the program SPCIM. Considering that high-grade squamous lesions are more frequently diagnosed than glandular lesions in histological outcome of women with these cytological diagnoses and the role of HPV in cervical carcinogenesis, this study was designed to analyze whether specific HPV genotypes may predict histological outcomes in women with glandular abnormalities in their cervical smears. Material and Methods: Of the 160 women included, 111 were diagnosed with AGC, 35 had both AGC and HSIL, while 14 women had AIS, in one case associated with HSIL. All women underwent colposcopic examination and biopsy was performed in 129/160 (80.6%). Thirty-one women (19.3%) were considered negative for neoplasia and scheduled for follow-up. All specimens were tested for 27 HPV genotypes by Roche's PCR-reverse line blot assay. The associations of specific HPV genotypes with cytological criteria were assessed through the calculation of odds ratios, with confidence intervals at 95% (95%CI).

Results: Histological diagnoses were either cervical intraepithelial neoplasia (CIN) or invasive carcinoma in 75 (58%) women, and negative for neoplasia in 54 (42% Objective: Cytopathic signs related to the human papillomavirus (HPV) infection are more frequently found in cervical intraepithelial neoplasia (CIN) 1; however, there are indications that the histological diagnoses of CIN2 and CIN3 can include areas of koilocytosis atypia and others signs. The objective of this study was to assess the frequency and the significance of the cytological criteria suggestive of HPV infection in the cervical smears of women with a histological diagnosis of CIN. Material and Methods: One hundred and sixty two women with abnormal cytopathologic findings and with a diagnosis of CIN including 46 cases of CIN 1, 42 cases of CIN 2 and 74 cases of CIN 3 were included. The cervical smears corresponding to each case were selected and analyzed by three observers. The divergences were resolved by consensus. The cytomorphological criteria analyzed included koilocytosis, spindle cells, cytomegaly, atypical parakeratosis and atypical metaplasia. For the statistical analysis, the odds ratio was used to assess the correlation between cytopathic signs and CIN grades. All analyses were performed using a 95% confidence interval (95% CI).

Results: Koilocytosis was found in 63% of the smears from women with a histopathological diagnosis of CIN 1. This sign was observed in 26.2% and 25.7% of smears of women with a diagnosis of CIN 2 and CIN 3, respectively. Cytomegaly also was frequent in cervical smears of women with histopathological diagnosis of CIN 1 (71.8%). On the other hand, spindle cells and atypical metaplasia were more frequent in women with CIN 2 and CIN 3 compared with women with the diagnosis of CIN 1. Atypical parakeratosis showed similar frequency in all grades of CIN diagnosis. Koilocytois and cytomegaly were inversely correlated with the diagnosis of CIN2 or CIN 3, with OR values respectively of 0.30 (95%CI 0.13-0.68) and 0.26 (95%CI 0.11-0.58). The others signs analyzed did not show any significant association. Conclusion: Koilocitisis and cytomegaly can provides good reassurance that a patient with atypical cervical smear have CIN 1. Objective: Cervical cancer screening is performed annually in Brazil with conventional cytology. Over the years, the usefulness of human papillomavirus (HPV) DNA testing has increased and has now become a valuable tool. Some of the specific uses of HPV detection are the triage of women with atypical squamous cells of undetermined significance (ASC-US) and an adjunct to cytology in routine cervical screening programs. A random model was applied for meta-analytical pooling and the influence of covariates on the HPV positivity rate was analyzed in comparison with conventional cytology, colposcopy and biopsy of the cervix. These methodological trials will allow us to establish criteria for screening programs to be used across Brazil. Objective: Cytology coupled with HPV status helps in screening women for cervical cancer. Prior experimental studies with conventional cytology for vulvar cancer have not produced meaningful results because of suboptimal sampling or processing. Brush sampling with liquid-based cytology (LBC), when coupled with HPV genotyping, may offer a better opportunity of screening women for vulvar dysplasia and cancer. Material and Methods: After institutional IRB approval, women from a vulvar clinic were recruited over a 1 year period. All vulvar lesions suspected for dysplasia were vigorously brushed and sampled in two PreservCyt Ò vials, one for LBC and the second for HPV genotyping, prior to biopsy for routine histology. Gentotyping was done with PCR using PGMY09/11 multiplex primers. Identical control samples were collected from clinically normal vulva for cytology and HPV typing without biopsy. Teams working on samples for cytology, genotyping or biopsy were blinded to each other's results or the patient's clinical status. Results were collated at the end of the study. Results: Fourteen samples from 10 women, age range 24-46 years, with biopsy-proven high grade VIN (HGVIN) qualified for this part of the study. Cytologic results were: Unsatisfactory (1), ASCUS (2), ASC-H (1), VIN 1 (1), VIN1-2 (7), HGVIN (2). Morphology assessment was based on criteria identified for cervical cytology. Hyperkeratosis was a constant feature of HPV infection of vulva. Abnormal cells were typically fewer than usually seen in similar lesions of cervix. All 13/14 satisfactory specimens for genotyping had HPV. HPV types in samples were: 1 type (3), 2 type (9), 3 types (0), 4 types (1). The genotypes were: Type 6 (1), 11 (1), 16 (5), 31 (1), 33 (1), 54 (3), 61 (2), 67 (2), 69 (1), 72 (1), 83 (4). Low risk HPV were always accompanied by high risk HPV (HRHPV). Type 18 was not identified in this cohort. Conclusion: Cytology, coupled with HPV typing, reliably identifies HGVIN. As previously reported, presence of Type 16 HPV can help identify patients more likely to develop squamous cell carcinoma. Presence of HPV types not covered by currently available HPV vaccines raises the possibility of incomplete protection, and a need for continuing monitoring of cases with vulvar dysplasia even in vaccinated women. Establishing consensus criteria for vulvar cytology will likely improve independent specificity for HGVIN. In the absence of genotypic evidence of HPV, the probability of a vulvar lesion being HPV-dependent HGVIN is negligible, and very unlikely in the absence of HRHPV. Larger controlled studies will help resolve hypotheses proposed in this study. Papanicolaou stain was used and the assessment of the slides was made by 3 independent cytopathologists for the SLE group and 2 for the control group.

Results: The prevalence of alterations in the SLE patients group was of 31.1% (11 ASC-US, 2 ASC-H, 5 LSIL, 6 HSIL, 1 AGUS) against 5% in the control group (4 ASC-US, 1 LSIL) (P = 0.000). No statistically significant association was found between the presence of alterations in pap smears and the variables DST history, first sexual intercourse at age £ 17, multiple partners ( ‡ 4), previous history of HPV, use of oral contraceptives, tobacco smoking, social class and cancer history in the family. Statistically significant association was found only in the variable group, in which a patient that belonged to the SLE group had an 8.2 higher chance (IC 95% = 2.7-24.9) of having alterations in the pap smear than a control group patient. Conclusion: In this study, 31.1% of the SLE women in intense immunosuppression presented cervical abnormality that was significally higher than the control group. These results suggest that SLE patients, when submitted to prolonged treatments with cytostatic agents, require special attention regarding the prevention of cervical cancer. Research shows the cause of cancer associated with sexually transmitted diseases (DST's) are important in understanding the different patterns of cancer observed in varying populations and age groups. The statistics in Brazil for epidemiologic infection of Chlamydia trachomatis (CT) are few. The infection is generally assymptomatic but can cause inflamatory pelvic disease, premature birth, neo-natal deaths in addition to other complications. The association of infection through human Papillomavirus (HPV) with CT is extremely high. This situation is particularly alarming in pregnant women who have low immunological resistance and for this reason are vulnerable. The purpose of our research was to evalute the frequency of infection by CT and relates this to the infection of high risk HPV within a populace of pregnant women. We studied 371 women with an average age of less than 25.4 years at an average gestation of 15.5 weeks. The Captura hybrid (CH2) for high risk HPV was positive in 50 (13.5%) of those examined. There was a significant relationship between high risk DNA-HPV and unstable marriage (OR = 2.61 (1.37-14.97) P = 0.003), CH2 positive for CT (OR = 3.03 (1.33-6.94) P = 0.009) and at the 25+age range (OR = 2.26 (1.09-4.71) P = 0.029). Of the 33 pregnant women with CT, 21 (63.3%) also present high and/or low risk DNA-HPV (P < 0.001). We concluded that the CT infection favors the entry and persistence of high risk HPV types, witch leads to viral integration. The traching of CT is an indispensable element in improving the qualitu of health of pregnant women. Objective: The aim of the study is to analyze the impact of the liquid based cytology with automated pre-screening and HPV testing on a population-based screening program with low incidence of cervical carcinoma. Results: A total of 290 HSIL cases were diagnosed, all of them histologically proved. 257 (88.6%) cases showed a viral load greater than 1 pg and were considered as HPV positive using HC-II method. 33 cases showed a viral load lower than 1 pg, and were considered as negative (11.3%). 10 of 33 HSIL-HCII-negative cases have been analyzed using PCR methods (Genomica, Madrid, Spain). The patients have a mean age of 47 years. All of them showed HSIL changes in the cytology sample. 40% showed moderate dysplasia and 60% showed changes of sereve dysplasia at the biopsy. The PCR results were as follows: five cases were consistently negative and five were positive four of them with high-risk virus (73, 53 or 16), and in one case there was not possible to determine the viral type.

We think it is necessary to investigate the presence of HPV in HSIL cases mainly in HSIL-HC-II negative ones. The HC-II negativity migth be explained for two different mechanisms: viral integration or E2 metilation. Furthermore there has been showed that some moderate or severe dysplasia cases could maintain the expression of adhesion molecules. In these cases the probability to obtain scraping atypical cells is theoretically lower and this fact might explain the negativity of HC-II, and probably he negativity of PCR. It must be considered too the possibility of carcinogenesis independently of HPV infection. Background: In addition to oncogenic Human papillomaviruses (HPV), several cofactors are needed in cervical carcinogenesis, but it is poorly understood, whether these HPV cofactors associated with incident (1) CIN1 are different from those required for progression to (2) CIN2 and (3) to CIN3.

Objectives: To gain further insights in the true biological differences between CIN1, CIN2 and CIN3, we assessed HPV cofactors increasing the risk of incident CIN1, CIN2, and CIN3. Study Design and Methods: Data of the NIS Cohort (n = 3187) and the LAMS Study (n = 12 114) were combined, and cofactors associated with progression to CIN1, CIN2 and CIN3 were analysed using multinomial logistic regression models with all covariates recorded at baseline HR-HPV-positive women (n = 1105), who represent a sub-cohort of all 1865 women prospectively followed-up in both studies. Results: Altogether, 90 (4.8%), 39 (2.1%) and 14 (1.4%) cases progressed to CIN1, CIN2, and CIN3, respectively. Baseline HR-HPV was the single most powerful predictor of incident CIN1, CIN2 and CIN3. When controlled for residual HPV confounding by analyzing HR-HPV positive women only, the risk profiles of incident CIN1, CIN2 and CIN3 were unique, i.e., completely different HPV cofactors were associated with progression to CIN1, CIN2 and CIN3 in univariate and multivariate analysis, irrespective whether non-progression, CIN1 or CIN2 was used as the reference outcome.

Conclusions: HPV cofactors associated with progression to CIN1, CIN2 and CIN3 display unique profiles, implicating genuine biological differences between the three CIN grades, which revisits the concept of combining CIN2 with CIN3 or (as also suggested) with CIN1.

patterns in cervix liquid-base specimens and relate them with hybrid capture 2 (HC 2) of high-risk HPV subtypes. Methodology: The authors performed p16INK4a and cytoactiv Ò HPV L1 immunocytochemistry in 98 liquid-base cervical samples, with cytologic diagnoses of atypical squamous cells of undetermined significance (ASC-US), atypical squamous cells cannot exclude high-grade squamous intraepithelial lesion (ASC-H), low squamous intraepithelial lesion (LSIL) and high-grade squamous intraepithelial lesion (HSIL); 1 case negative for intraepithelial lesion or malignancy as an internal control. The results of immunocytochemistry were compared and associated to results of HC. Conclusion: The expression patterns of L1 screening antibody were more easier to identify in pap slides than p16 antibody, probably due to the fact that the last one can be found in metaplastic and reactive cells besides dysplastic ones. The L1 detection in association with p16 as a second biomarker seems to be an useful tool in the behaviour prediction of dysplastic lesions associated with HPV infection. Objective: The recurrence of primary oral's tumor after treatment has been reported as 25-45%. The tumoral recurrence, as well as the interval in which it takes place has a good correlated prognostic value. The routine use of smears for monitoring and diagnosis of the recurrences of buccal cancer can have a positive impact in the patient prognosis. Materials and Methods: Nineteen cytologic smears were taken from oral mucosa of patients in attendance after oral carcinoma surgery. The chosen area was the site with surgical scar or in places with clinical alterations. A triple smear was carried out using a cytobrush (Koplast Comercial Industrial Ltda., Brazil), a metal and a woode spatula. The smear were transferred to clean, dry glass slides and fixed by the polyethylenoglycol and processed for cytogical assessment using the Papanicolaou staining procedure. The Bethesda guidelines was applied for the diagnosis.

Results: In five cases clinically classified as normal, the smear was obtained from surgical scar. In one of them atypical cells of indeterminate meaning (ASCUS) were found and the patient stayed in observation. In six cases with typicals leucoplakias, the cytological smear showed inflammatory features. In five cases clinically classified as erythroplakias, there was one positive to maligne lesion with histological confirmation. There was two ulcerated lesions and one with clinical tumoral characteristics, but none presented compatible alterations with recurrence, in spite of the clinical suspicion.

Conclusion: The use of the cytologic smear is a good method for diagnosis of recurrences for patients in attendance after oral carcinoma surgery treated surgically for ablation oral carcinoma. The cytologic study of the cells of the oral cavity is simple, quick, not aggressive and relatively painless method. The cytologic examination is well accepted by patients and is efficient for analysis of suspect injuries and for attendance of malignant confirmed injuries after treatment. Columnar cell variant of papillary thyroid carcinoma (CCV-PTC) is a rare and aggressive tumor with widespread nodal and visceral metastasis. Histologically, the tumor is composed of frequent papillae lined by elongated cells whose nuclei are stratified and appear to be rich in chromatin. In this report we present the fine-needle aspiration cytologic findings of two cases of CCV-PTC with cytohistologic correlation. Smears showed tall columnar cells in monolayered, three-dimensional sheets and frequent papillary clusters. Several of the fragments showed columnar cells with nuclear pseudostratification, resembling that seen in colonic adenomas. Nuclei were oval or elongated and monomorphic. The classic nuclear features of PTC -including optical clearing, grooves, and inclusions-were rarely seen. The resected thyroid tissues revealed a columnar cell variant of PTC. The differential diagnosis of CCV-PTC includes tall cell variant of PTC, medullary carcinoma of thyroid and metastatic colorectal adenocarcinoma. Tall cell variant of PTC can be distinguished from CCV-PC by the oxyphilia of the tumor cells and the absence of nuclear pseudostratification. In other two tumors, immunohistochemical stains for calcitonin and thyroglobulin play an important role in difficult cases. The cytologic features of CCV-PTC are at variance with those of classic PTC and are elusive in fine needle aspiration cytology. The definite cytologic diagnosis of this aggressive variant is very important in designing the best modality of treatment. Solid papillary carcinoma of the breast is a subtype of intraductal carcinoma, which occurs in older women and usually has a favor-able prognosis. It is primarily intraductal, but also is often associated with invasive carcinoma. Recently, we encountered a case of solid papillary carcinoma and herein report the cytologic findings. A 89-year-old Japanese woman presented with a left breast mass, the fine-needle aspirates of which showed moderately cellular smears of polygonal to cuboidal tumor cells arranged in the form of papillary clusters with thin, fibrovascular cores. However, cellular cohesiveness was variable and dispersed single tumor cells or small clusters were also observed. The nuclei were round to oval, eccentrically located nuclei, showing plasmacytoid appearance, and have fine chromatin and inconspicuous nucleoli. The cytoplasm of the tumor cells was abundant and somewhat eosinophilic. There was neither necrosis nor naked nuclei. Mucinous material was not evident. Microscopic examination showed a well-circumscribed tumor (1.8 · 1.8 · 1.3 cm in size). The tumor cell proliferation was essentially solid in architecture. The tumor was composed by a uniform population of small to medium-sized, ovoid or spindle-shaped epithelial cells with round to ovoid nuclei with finely dispersed chromatin. A few fibrovascular cores, resembling the configuration of a palisade or pseudorosette, were observed. Mitotic figures were rare and no comedo necrosis was observed. Immunohistochemical studies showed focal positivity for synaptophysin. ER and PgR were positive, but overexpression of c-erb-B2. oncoprotein was negative. The patient was well at the follow up examination 1 year after operation. Objectives: The disease of nodular thyroid is frequently observed (25-40% in ultrasonographic evaluations and autopsies) and has difficulties in clinical practice. It is a common and frequently benign disease having incidence which increases with age, and is increased in women, in people with iodine deficiency, and after radiation exposure. Materials and Methods: We aimed to report the retrospective analysis of diagnostic results of fine-needle aspiration cytology of thyroid gland (FNAC-t) is performed from the same surgeon in the first year of a peripheral state hospital. In the present study, 52 fine-needle aspiration cytology of the thyroid gland is performed at Department of General Surgery, Ankara Golbasi Hasvak State Hospital between the dates of October 2006 and February 2008 which were evaluated retrospectively and classified as non-diagnostic, benign, indeterminate, and malignant. Results and Conclusion: A great majority of our cases had benign characteristics and the generally of the benign cases were colloidal nodular cytology. Only two indeterminate cases were detected among the 52 cases and postoperatively, one of them had been exhibited thyroidal malignity as papillary thyroidal carcinoma. According to terminology in Bethesda; diagnosis of FNAC-t classified as non-diagnostic, benign, follicular lesions of undetermined significance, suspicious for follicular neoplasm/suspicious for Hurthle cell neoplasm, suspicious for malignancy, and malignant. Anew aspiration for non-diagnostic, follow up for benign (malignity risk: < 1%), a new aspiration for follicular lesions of undetermined significance (malignity risk:~1%), lobectomy for suspicious for follicular neoplasm (malignity risk: 20-30%), lobectomy for suspicious for Hurthle cell neoplasm (malignity risk: 20-45%), lobectomy or thyroidectomy for suspicious for malignity (malignity risk: 60-75%), thyroidectomy for malignity (malignity risk: 97-99%) is suggested as clinical algorhythm of recommended Bethesda terminology. The aim of the study was to present a retrospective analysis of FNAC-t of the patients admitting to the unique hospital of a county in the interval of the first year of the hospital. Objective: We reported useful method using glucomannan for the distinction between malignant mesothelioma and ovarian carcinoma using cell block in ascites. The present study was established using transmission electron microscopy (TEM) from culture cell. Materials and Methods: The specimens were taken from ascites of a patient with definitively diagnosed ovarian carcinoma. The techniques employed in fix, centrifuge, glucomannan, and paraffinembedded block were essentially the same as described earlier.

TEM method was as follows; (i) the samples of was subjected to centrifugation at 1500 rpm for 5 minutes and the supernatant;

(ii) they were fixed in 1% paraformaldehyde and 1% osmium tetroxide; (iii).they were treated successively with each step of alcohol (50-100%) for 10 min. It puts it in the glcomannan formalin liquid in the specimen; (iv) glcomannan hardens if the specimen is put in the methanol for two hours. Then with a mixture of epoxy resins/QY1 (6 : 4), and pure epoxy resins for 60 minutes. The samples were sectioned by an ultracut and the sections were subjected to electron staining with uranium acetate and lead citrate for three min respectively. They were observed and photographed under a JEM-1010. Objective: We evaluated the follow up of 180 patients with superficial bladder cell carcinoma (T1-grade 2-3) after TUR-B and intravesical chemotherapy for the detection of recurrences and progression of these tumors first diagnosed by cytology followed by cystoscopy and biopsies. We also studied the impressive characteristic cytomorphologic changes due to the response of the chemotherapeutic agents. These tumors are multiple, multifocal and of high recurrence rate. Materials and Methods: We reviewed 180 patients aged 43-92 years with superficial urothelial bladder cell Ca grade 2-3 (T1). The mean follow up was 3 years. We evaluated over 1850 urine specimens (voided urine and bladder washings) after TUR-B and intravesical instillations of combined chemotherapy and BCG according to the protocols. All urine samples (voided urine and washings) were double cytocentrifugated and stained by two different staining techniques: Papanicolaou Stain and Rapid stain Hematoxylin -Eosin (HE) -first application in urine.

Results: Intravesical chemotherapy after TUR-B reduces the tumor recurrences (35-65%) and also induces specific cytomorphologic cell changes such as: impressive enlarged multinucleated urothelial cells with foamy vacuolated cytoplasm, with irregular dark nuclei with prominent nucleoli. By the application of the rapid Stain HE first applied in urine cytology, we obtained a priority of cellularity (over 70%) and better diagnostic results. Conclusion: The diagnostic accuracy by cytology of monitoring patients underwent TUR-B and combined chemotherapy arises by an experienced cytopathologist from 86-92%. The cytology follow up gives also important information to the Urologists about the response or not (MDR) to the applied intravesical chemotherapy. These patients have to be followed up longlife by cytology and cystoscopy every six months for possible recurrences. Objective: At our laboratory we examine approximately 1075 mamma FNA cytology specimens per year including palpable and non-palpable masses from a cluster of 4 hospitals. We describe our practice and experience over the past 5 years using a standardized protocol for increased quality. Materials and Methods: We classified each specimen into one of five categories on a scale of likely subsequent histological outcome: benign (B), uncertain benign (UB), uncertain malignant (UM), malignant (M), no diagnosis (ND). At regular intervals we analysed all cases and re-assed each one with a two step or more discrepancy with final histology in order to gain knowledge and expertise.

Results: About 40% of FNA's had subsequent follow up histology. We report an overall sensitivity of 78.4% including ND FNA, sensitivity of 90.1%, specificity of 95.7%, positive predictive value of 98.0% and negative predictive value of 80.3%. In practice, clinicalpathological correlation improved the real value of FNA further as it supported the clinical triple test. Pitfalls and areas of heightened scrutiny were: adjacent lesions punctured in one specimen, pregnancy changes, atypia after therapy, granular cell tumor, low grade neoplasia, metaplastic carcinoma, in-situ, cystic and papillary tumors. Conclusion: Although the trend in healthcare is towards initial histological core biopsy of mammary lesions, we find that FNA contributes to an important triage step in patient care and can guide towards an optimal cytology appreciation of difficult lesions if done in a well monitored setting with quality control including follow up with a clinical triple test setting. We emphasize a balanced report which leads to a best subsequent clinical approach and step wise management tailored to each case while using the five above categories to limit diversity in reporting. FNA remains a fast and easy test so that resources can be well guided. Introduction: Adenoid cystic carcinomas (ACC) of the lung and bronchus comprise less than 1% of all pulmonary tumors. They are derived from bronchial mucous glands and therefore tumor is located in the trachea, bronchus main stem or lobar bronchi. The diagnostic cells are more likely to be found in bronchial brushing specimens than in sputum because these neoplasms are covered by intact mucosa. Patients are more prone to develop local recurrence because of difficulty obtaining clear margin. Poor prognosis is related to stage of tumor at the time of diagnosis, the presence of positive margin and a solid cellular growth pattern. Regional lymph node and systemic metastases are seen in approximately 20% and 40% of cases.

Case report: The patient was a 60 year-old woman who presented with shortness of breath, cough and haemoptysis. Radiographs showed a centrally located mass and bronchoscopy showed a tumor affecting main bronchus in the left lower lobe. Material from both needle aspirates and bronchial brushings were obtained. With the Papanicolaou stain, there were clusters of small, uniform cells with scanty cytoplasm and monotonous dark nuclei arranged around spheres of the blue hyaline material. In core biopsy, the neoplastic cells were uniform and small, arranged in cribriform arrays and solid nests. The patient underwent resection of the left lower lobe and was reoperated with left hemipulmectomi because of unclear margins. Histologic section showed a typical pattern with ACC. A five-year-follow-up shows no recurrence or metastasis of the tumor.

The cytologic picure is characteristic, but because this is a rare condition and the nuclear atypia is minimal, the diagnosis might be overlooked. ACC and other salivary-gland type tumors must be kept in mind when performing bronchial cytologic examinations from centrally located lung tumors. The development of synchronous malignant mesothelioma with adenocarcinomas is rarely being reported. The presented case, concerns a 64-year-old man, with known chronic obstructive disease of the lung, who was admitted to our hospital with left pleural effusion. The patient had no history of asbestos exposure. The chest CT revealed left pleura thickening. No malignancy in the lung parenchyma was seen. The cytological examination of pleural effusion suggested the diagnosis of malignant mesothelioma. Thoracotomy was performed and biopsies were obtained. The histological and immunohistochemical analysis (TTF-1-, CEA-, WT-1+, CK5/6+, Vimentin+) confirmed the cytological diagnosis. The patient underwent three cycles of chemotherapy and showed significant clinical and radiological improvement. He presented 4 months later complaining for dyspnea and cough. Chest CT revealed a large solid lesion occupying most of the right lung parenchyma. Infiltration of the mediastinum and multiple small nodules, in the left lung, were also noted. Bronchoscopy was performed and washings and post bronchoscopic sputum samples were obtained. The cytological smears revealed a large number of malignant cells mainly of papillary configuration. The neoplastic cells demonstrated foamy cytoplasm, mostly eccentric nucleus and prominent nucleoli. Immunohistochemical studies, on cell block preparations of the bronchial washing material, revealed that the neoplastic cells were positive for CEA, TTF-1, CK5, CK7 and negative for WT-1-, calretinin, vimentin, CK5-6, thrombomodulin, HMBE-1 and CK20. Thus, the diagnosis of pulmonary adenocarcinoma was established. The patient died 2 months later. The present report calls attention to the likelihood of a simultaneous occurrence of pulmonary adenocarcinoma and malignant mesothelioma. Introduction: The importance of Her2 analysis in the management of breast cancer is well documented. FISH is regarded as the golden standard for the Her2 test. Chromogenic in situ hybridization test for Her2 (CISH) was previously tested on histology as well as on smeared cytology. The aim of this study was to discover the diagnostic feasibility of CISH on LBC prepared as SurePathä specimens, which we believe will save time and manpower in our laboratory. Materials and Methods: For this comparative study, we had unfixed breast tumors at our disposal. The fresh cut-surface of each tumor was brushed and the brush head was dumped into the Sure-Path vial. Formalin fixed and paraffin embedded sections were then taken from the same cut-surface. Two unstained slides were made from both the LBC and histological specimen. The LBC slides were prepared on PrepStainä non-gyn program. One slide from histological as well as LBC specimens were hybridized on Dako's Hybridizer followed by manual CISH staining. Immunostaining for Her2 was performed on the remaining two slides using Ventana's BenchMark-XT.

Results: Stainings for Her2 on both histological and LBC specimens were successful as were our CISH on histological specimens. The results on these tests were consistent with each other. Although the results of the CHIS test on LBC were successful, they varied from day to day, more so than the CISH test on histological specimens. We have also noticed that the more amplified a cell was the easier we had to get an optimal result on LBC slides. Aim: Cytological study of submit unfixed lymph node tissue currently has several objectives, such as diagnosis, confirming suspected FNA findings or, assessing the representativeness of tumor tissue, for the tumor bank. In this study, we perform a morphologic evaluation of cytological samples obtained in fresh lymph nodes to determine the usefulness and advantages of these techniques in daily practice. Materials and Methods: We reviewed 69 fresh lymph node samples. Sentinel nodes and lymphadenectomies for oncological disease were excluded. Among the total, 42 specimens were submitted to evaluation for inclusion in the tumor bank and the remaining to establish the diagnosis. Thirty-three (33) cases had a previous fine needle aspiration (FNA) . In all samples, tissue imprints and scrape cytology of the cut surface with Diff-Quick staining were performed; flow cytometry was additionally carried out with lymphomas.

Results: Among the 42 samples submitted for the tumor bank, 5 (12%) were not diagnosed with fresh tissue cytology: two atypical hyperplasia, one dermatopathic lymphadenopathy, and two reactive adenitis. In the five metastatic cases, the diagnosis was established by cytological evaluation of fresh material in all cases. Of the 33 with a prior FNA, eight had a diagnosis of suspected malignancy with this technique, and on fresh tissue cytology, four were lymphomas and three benign; the last sample was atypical hyperplasia, diagnosed in paraffin-embedded sections. Among the lymphomas diagnosed by FNA (7 cases), five were confirmed by fresh tissue assessment and two were found to be false-positives (1 granulomatous lymphadenitis and one atypical hyperplasia).

Comments: In comparison with tissue imprints, scrape cytology preparations present some advantages in both the quality and quantity of cells. Evaluation of cytological smears are simpler and faster (2 minutes) than frozen sections. Cytological assessment of these preparations is helpful in inconclusive FNA, although the diagnosis of atypical hyperplasia may be more accurate in paraffin sections. Scrape cytology is a useful quality control method for lymphoma samples destined for the tumor. Conclusions: The most common cause of diagnostic error was sampling of inadequate or non-representative material because of necrosis or cystic degeneration of the tumors, especially in astrocytic tumors. The most important advantages of this method are the preservation of cytologic features which enables a rapid intraoperative diagnosis even when frozen section is not possible due to the size and consistency of tissue samples, the speed and simplicity of the process and the possibility of tissue preservation for further examination, e.g. histological or immunohistological. Therefore, the cytologic examination is an extremely accurate method for the intraoperative diagnosis of CNS tumors and a necessary complement of histology and immunohistochemistry. [2006] [2007] [2008] . The samples were obtained by fine needle aspiration (FNA) or by brush from ulcerated lesions and subsequently were smeared on slides and stained with rapid Giemsa (Hemacolor) and Papanicolaou stains. Results: The cytological diagnosis revealed: four cases of giant-cell epoulis,four chronic obstructive sialadenites (sublingually), three Non-Hodgkin lymphomas (lingual, tonsillar, Waldayer ring), 11 lingual squamous carcinomas,one squamous carcinoma remission (sublingual mass), one keratinized nasopharynx carcinoma (metastatic to tonsil), one primary tonsillar squamous carcinoma, two secondary small cell lung carcinomas in tonsil and buccal mucosa, two minor salivary gland mixed tumors (sublingual and palatal), 12 squamous carcinomas (of the oral mucosa or floor, the palate, or the pharynx), three ameloblastomas (maxillar and mandibular), one sublingual salivary adenoid cystic carcinoma and one minor salivary gland oncocytoma of the palate. Surgical excision and histologic confirmation followed in 25 cases.

Conclusion: The majority of the oral cavity tumors refers primarily to high or low differentiation squamous carcinomas and secondarily to salivary gland tumors. The cytologic examination is a rapid, simple and specific method for the diagnosis and classification of oral cavity lesions (primary or not) and a valuable tool for the design and planning of further patient treatment. . For cytological analysis two to four smears from each aspirate and cytospins from the rinsing product of the needle were stained by MGG. In order to identify the source of foreign material in some slide, all cytological samples were re-examined blindly. The contamination was assessed by a semi-quantitative score. Mineral analysis of the rinsing product of needles was performed using a transmission electron microscope equipped with an energy dispersive X-ray spectrometer.

Results: Extra-cellular black particles different from dusts or anthracosis were identified only in the 57 samples from EBUS-TBNA aspiration. No difference was observed between five batches of needles. The contamination score and the number of passes during the procedure was correlated (P = 0.035). Mineral analysis of the washing products showed no particle with conventional needles while metallic compounds of iron, titanium, nickel and chromium were found with EBUS dedicated needle. Conclusion: The use of a dedicated EBUS-TBNA needle is associated with metallic particle release probably by friction between stylet and needle. These particles could be miss interpreted as anthracosis and may interfere with cytological examination. Objective: Liquid based cytology preparation of thin layer slides has improved morphological evaluation in diagnostic cytology and allowed immunocytochemical and molecular analyses on the fixed cells. Protein expression in cultured cells is usually evaluated by immunofluorescence or Western blot. Immunofluorescent cells, however, can only be stored for a week, while western blot is laborious, and requires a high cell number. We utilized liquid based cytology to study the expression of nuclear proteins in cultured cells.

The IL-7-dependent murine cells D1 and human peripheral blood (PBL) mononuclear cells cultured in IL-7 and starved for 4 and 32 hours, respectively, were stimulated with IL-7 (50 ng/ml) for 15 minutes to induce phosphorylation of the transcription factor g/ml) for 72 hours to lSTAT-5 (pSTAT-5). PBL cells were stimulated with PHA (two induce expression of Ki67 antigen. Cells (4 · 106) were washed once and fixed in Preserv-Cyt Ò . Results: In D1 cells pSTAT-5 immunoreactivity was negative on unstimulated cells and positive on IL-7 stimulated cells. Immunoreactivity and cell morphology were clearly appreciated 24 hours, 1 and 4 months after fixation. In PBL cells pSTAT 5 immunoreactivity after IL-7 stimulation and Ki67 immunoreactivity after PHA stimulation were appreciated 1 month after fixation, but the cell cytoplasm was damaged. Omission of antigen retrieval greatly improved the morphology of PBL cells while pSTAT-5 and KI67 We got a fine needle aspiration biopsy from a lesion of the 84 years old pacient with suspect tumor of the epigastrium to the examination on many slides and two cytoblocks.

Results: On the slides were haemolytic blood on a backround, mostly elongated cellules, coreless, unnoticed clusters and in cythoblocks were found spindle shaped elements with elongated nucleuses with oedematous saturation. Imunohistochemical examination was CK AE1/AE3 and S100 negative, Actin, H-Caldesmon, CD34, and CD117 positive. The diagnosis from cytology was derived as GIST. Gastrointestinal stromal tumors (GIST) represent most of the mesenchimal neoplasms arising in this setting. They are more frequent in the stomach and its biological behavior varies from benign small-middle size tumors to true malignant sarcomas. They usually present as solid and firm masses, although some may show focal cystic changes secondary to degenerative processes, necrosis, hemorrhage or due to treatment with thyrosin-kinase inhibitors. Nevertheless, frank cystic degeneration is infrequent in GIST, raising a differential diagnosis other than the usual that includes vascular lesions and pancreatic or GI cystic and pseudocystic tumors.Ecoendoscopy-guided fine-needle aspiration (EUS-FNA) is a useful and secure technique in the diagnosis of intra and extramural tumors of the GI tract, not accessible by means of simple endoscopic biopsy, with sensitivity, specificity and diagnostic accuracy figures superior to 90%. The rapid evaluation of the aspirated material and the use of complementary immunocitochemistry stains helps to obtain outstanding results with this technique. We present two GIST cases detected as subepithelial solid-cystic gastric tumors with a difficult radiologic characterization. Both cases were diagnosed by means of EUS-FNA and, after surgical resection, the diagnosis was histologically confirmed. Systemic lupus erythematosus (SLE) is a chronic, inflammatory disease that can affect several organs of the body. It is an immunemediated disorder characterized by a multisystemic microvascular inflammation with the production of autoantibodies. SLE frequently affects women during childbearing age and the use of exogenous hormones has been associated with lupus onset and flares, suggesting a hormonal role in the pathogenesis of the disease. Several studies have also investigated the role of infectious etiologies such as Epstein-Barr virus (EBV) that may also contribute to autoimmunity. Oral ulcers may be noted in 20% to 30% of the patients and comprise one of the diagnostic criteria. Other oral manifestations associated with immunosuppressed therapy may occur. The aim of this study was to investigate the frequency of oral lesions in LES patients and the importance of the oral cytology in the diagnosis of clinical and subclinical oral lesions. A prospective study was performed in 155 patients with diagnosis of SLE. Incisional biopsies and scraping were carried out in the lesions and at the lateral borders of the tongue respectively. It was identified 13 oral lesions in 10 patients. A biopsy was performed in eight lesions and three of them were suggestive of SLE manifestations. Cytopathologic exams were accomplished in 110 patients. It was observed 15 (13.7%) cases of subclinical oral candidiasis and four (3.7%) cases of oral hairy leukoplakia (one clinical and three in subclinical forms). The prevalence of the oral lesions in patients with SLE is low (6.5%). Besides, it was observed the importance of cytopathology exam in the detection of infectious lesions that may be associated with immunosuppression of SLE. The oral cancer is one of the most common cancers worldwide and constitutes a health problem. A key factor in the lack of improvement in prognosis over the years is the fact that a significant proportion of oral squamous cell carcinoma are only diagnosed or treated when they reach an advanced stage. Cytological study of oral cells is a non aggressive technique that is well accepted by patients. However, its usage has been limited due to poor sensitivity and specificity in diagnosing oral cancer. Objective: FNA cytology is the principal method for the investigation of thyroid nodules with a worldwide acceptance. However, the sensitivity of the method needs improvement as there are always a few ambiguous cases, in which even in the more experienced institution, a 'suspicious' diagnosis will be rendered. In the present study we evaluate the contribution of galectin-3, cytokeratin 19 and HBME-1 expression in establishing an accurate diagnosis. Materials and Methods: Fourty thyroid FNAs performed on surgically excised thyroid nodules were included in the study. The material consisted of 15 papillary carcinomas, two follicular carcinomas, two follicular adenomas, one Hurthle cell adenoma, 18 nodular goiters and three cases of Hashimoto's thyroiditis. Conventional and thinprep cytology diagnosis and the immunomarkers' (galectin-3, CK19 and HBME-1) expression were correlated with histology.

Results: 14/15 papillary carcinomas (PC), 5/5 follicular neoplasms (FN), 21/21 benign lesions (BL) were diagnosed by cytology, conventional and thinprep. HBME-1 positive staining was seen in 10/ 15 PC, 1/2 FCa, 0/3 FAdenomas, 0/21 BL. Galectin-3 positive staining was present in 12/15 PC, 1/2FCa, 2/3 FA, 4/21 BL. CK19 positive staining was observed in 11/15PC, 2/2 FCa, 1/3 FA, 3/21 BL. Thus, sensitivity for papillary Ca was for cytology 93.3%, HBME-1 66.67%, Gal-3 80%, CK19 73.3% and for benign lesions 100% for cytology and HBME-1 non-staining. It is interesting that every neoplasm diagnosed by cytology was stained at least by one of the tested markers. The papillary carcinoma missed by cytology was also negative for all the markers, as it was a microcarcinoma that FNA failed to sample. Conclusion: The combination of galectin-3, CK19 and HBME-1 expression can assist morphology and provide a more accurate diagnosis in thyroid FNA cytology. Introduction: Lymphoma arising within the oral cavity accounts for less than 5% of all oral malignancies.Waldeyer's ring is the most common site of lymphomas involving the oral region and tonsil is the most prevalent site of the involvement 2.7%). Cervical lymph node enlargement is often an early clinical manifestation of the disease. FNA plays an important role in differentiating inflammatory conditions from malignant neoplasms and represents an accurate method in establishing diagnosis of Waldeyer's ring lymphomas (sensitivity 93.10% and positive predictive value 100%). We present two cases of tonsilar lymphomas, diagnosed by FNA cytology of enlarged cervical lymph node. Case presentation: Case 1: female, 50 years-old presented with left tonsil enlargement and left cervical lymph node enlargement (6 cm). Case 2: male, 40 years-old presented with sore throat and right cervical lymph node enlargement (4 cm Objective: The purpose of this study was to clarify sequential changes in the cytological features of pleural fluids in sarcomatoid mesothelioma.

Materials and Methods: Pleural fluid cytology specimens were obtained from a patient with a final diagnosis of sarcomatoid mesothelioma of the right pleura. The patient was an 82-year-old man with silicosis and a history of acute myocardial infarction 8 years previously, which precluded any invasive examination or treatment. He died of respiratory failure after a clinical course of approximately 7 months. During his seven admissions, thoracentesis was performed five times, and Papanicolau-or Giemsa-stained smear preparations were made on admission and 2, 3, and 4 months later. At the same time, H&E-stained cell blocks including immunohistochemical examination were prepared. Results and Conclusion: Cytological smear preparations initially showed small, scattered, piled-up, or sheet-like clusters of slightly atypical epithelioid cells, accompanied by many lymphocytes and macrophages. Subsequently, these epithelioid cell clusters gradually disappeared, isolated atypical large cells with prominent nucleoli appeared in small numbers, and, finally, these cells increased in number and their degree of atypia. Immunohistochemically, the atypical cells were positive for the epithelial cell markers and the mesenchymal marker, negative for the adenocarcinoma markers, and partially positive for the mesothelioma markers. These findings suggest that a low number of small slightly atypical epithelioid cell clusters and isolated atypical large cells appear in the early stage of sarcomatoid pleural mesothelioma. Fine needle aspiration cytology of germ cell tumours at extragonadal locations Objective: The objective of this study was to evaluate and describe the various cytologic features of extragonadal germ cell tumours, both primary and metastatic, in fine needle aspiration cytology. Materials and Methods: Aspirates of all extragonadal germ cell tumours diagnosed over a 11-year period were retrieved. Clinical details, including age, sex, site of aspiration and a prior history of gonadal tumour were recorded. The cytologic smears were reviewed and various features, including cellularity, cell patterns, cytoplasmic and nuclear features, background characteristics and any additional features were noted. An attempt was also made to sub-classify the germ cell tumours in all cases. Corresponding histologic sections were also reviewed, wherever available. Results: Of the 88 cases retrieved, 57 with adequate cytologic material and clinical data were finally analyzed. Seminoma/dysgerminoma showed dyscohesive cells with well-defined finely vacuolated cytoplasm, central nuclei with prominent nucleoli. The characteristic 'tigroid' background was seen in only a minority of these cases. Yolk sac tumour was recognized by papillary fragments of tumor cells with metachromatic basement membrane-like material in the cell clusters. Embryonal carcinoma, in contrast to seminoma and yolk sac tumour, did not have characteristic morphologic features. Mixed germ cell tumours were difficult to recognize as such due to frequent sampling bias in cytology.

Conclusions: Fine needle aspiration cytology is a reliable diagnostic tool for germ cell tumours at extragonadal sites, even in rare locations and for patients where metastatic tumour is the first clinical presentation. The cytopathologists need to remember the diagnostic problem posed by immature teratoma and mixed germ cell tumours due to sampling error. Also, the list of differential diagnoses at extragonadal sites is wider than at gonadal sites. Background: Compared to epithelial tumors metastases of melanoma to bone marrow are rare. Case report: A 46-year-old man was admitted to hospital with fever and hepatosplenomegaly. A blood cell count showed anemia, thrombocytopenia and normal differential leucocyte count. Anisocytosis, hypochromia and few nucleated red blood cells was found in peripheral bood smear. Abdominal CT scan revealed liver enlargement. Fine needle bone marrow aspiration (sternum) and bone marrow biopsy (pelvis) were performed. Results: Fine needle aspiration smears were cellular and showed replacement of normal marrow elements with large abnormal cells and macrofages with brownish cytoplasmic pigmentation. Immunocytochemicaly tumor cells were positive for melan -A and S-100 and negative for LCA and BerEP4. Bone marrow biopsy showed numerous tumor cells with hiperchromatic nuclei and cytoplasmic pigment. Immunohistochemicaly these cells were positive for melan-A, HMB-45, S-100 and negative for AE1/AE3 and synaptophysin. A diagnosis of malignant melanoma with bone marrow involvement was made. Patient passed away 10 days from admission. Autopsy revealed metastases to several visceral organs, and no tumor primary site. Conclusion: Metastatic involvement of bone marrow by melanoma of unkown primary has been reported rarely. The diagnosis is relatively easy, if melanin is found in the tumors cells and macrofages. Immunohistochemical staining is important to confirme definitive diagnosis. Objective: The aim of this study was to evaluate the significance of selected immunocytochemical stains in differentiating metastatic adenocarcinomas infiltrating the pleura or the peritoneum from benign reactive mesothelial cell proliferation. Methods: The material encompassed 84 cases of metastatic adenocarcinomas in serous effusions and 76 cases of reactive mesothelial cells. A panel of immunocytochemical stains was used in this study which included HBME-1, Calretinin, Ber EP4, B72.3, CA 125 and TTF-1 antibodies. The processing of the cytologic material was accomplished according to the Thin Prep technique. Results: Among the antibodies used in the study, anti-HBME-1 had the highest sensitivity (75.3%) but lowest specificity (76.2%). Staining for calretinin showed high specificity (97%), with moderate sensitivity 62%. When membrane staining was evaluated, the sensitivity of the markers studied in detecting malignant cells was as follows: Ber EP4: 75%, B72, 3: 76%, CA125: 84%. Membrane positivity for Ber EP4 and B72, 3, was not detected in reactive mesothelial cells. However, membranous staining in mesothelial cells was evident in 28% of cases with the use of CA 125.When Ber EP4 and B72,3, staining results in cancer cells were combined, the sensitivity was higher (Ber EP4 + B72,3: 89%).Staining for TTF-1 showed the highest specificity (100%) for adenocarcinomas of pulmonary origin, with moderate sensitivity (70%). Conclusion: The detection of malignant cells in effusions is facilitated by the use of immunocytochemistry using a wide panel of antibodies. The best combination of sensitivity and specificity was found for B72, 3, Ber EP4 and TTF-1 and for Calretinin and HBME-1. Background: Histopathologic classification of lung carcinoma is important, as prognostic factor and in the evaluation of treatment modalities. So, it is necessary to define in almost each case whether a carcinoma is a small or non-small cell carcinoma (adenocarcinoma, squamous or large cell carcinoma). Objective: In this study, we sought to validate the importance of TTF-1, p63, CD56, CK5/6 and CK7, as an antibody panel, for differentiating lung carcinomas. Methods: Bronchoscopic samples (washing, brushing, TBNA), from 358 lung carcinomas (70 small cell lung carcinomas, 126 squamous cell carcinomas, 142 adenocarcinomas and 20 large cell carcinomas), were evaluated. The cytologic material was processed using the Thin Prep technique. Immunocytochemistry was performed using specific monoclonal antibodies.

Results: All small cell lung carcinomas were positive for CD56, while 92.8% (65/70) of them showed diffuse moderate or strong staining for TTF-1, with no staining for CK5/6 and p63. In contrast, 95.2% (120/126) of squamous cell carcinomas manifested opposite immunoreactivities, with diffuse moderate or strong staining for CK5/6 and p63 and no staining for TTF-1 and CD56. Lung adenocarcinomas were consistently positive for CK7 and TTF-1 and negative for CD56. Only a small number of them demonstrated positivity to CK5/6 and p63. More than 50% of large cell carcinomas were of the same immunophenotype as adenocarcinomas. Conclusion: The immunophenotypes of the main histologic types of lung carcinoma are stable and highly reproducible. However, because of considerable overlapping, immunophenotyping should not be used alone for histopathologic classification of lung cancer, but only as an adjunct to light microscopy. Methods: Forty-two urine samples collected from immunosuppressed patients were processed using the ThinPrep method and analyzed by Papanicolaou stain and immunocytochemistry with a JC/BK virus-specific monoclonal antibody. Results: The morphologic sign of the (re) activation of polyomaviruses was the detection of typical intranuclear viral inclusion bearing epithelial cells, so-called 'decoy cells', in the urine. Overall, polyomavirus was positive in 15 of 42 urine samples (357%). Of 15 samples, 12 (80%) were positive by both methods, while 3 (20%) were positive by immunocytochemistry only. Also, immunocytochemistry detected more infected cells in samples with few positive urothelial cells, which would have gone undetected if only Papanicolaou staining had been used, as screening method. Conclusion: The main advantage of immunocytochemistry is that it allowed confirmation of polyomavirus infection diagnosis in urine samples. Urine samples testing for polyomavirus by both techniques will improve diagnosis in asymptomatic patients, allowing early therapeutic intervention and a better clinical outcome. Introduction: Gene expression studies related to cancer diagnosis and treatment are becoming more important. Housekeeping genes that are absolutely reliable are essential for these studies to normalize gene expression. An incorrect choice of housekeeping genes leads to interpretation errors of experimental results including evaluation and quantification of pathological gene expression. Materials and Methods: We examined (a) the degree of regulation of GAPDH expression in human glioblastoma cells under hypoxic conditions in vitro in comparison to other housekeeping genes like beta -actin, serving as experimental loading controls, (b) the potential use of GAPDH as a target for tumor therapeutic approaches and (c) differences in GAPDH expression between differnt WHO-grades of astrocytic tumors. GAPDH and beta -actin expression was comparatively examined in vivo in human lowgrade astrocytoma (WHO grade 2) and glioblastoma (WHO grade 4) on both, protein and mRNA level, by Western blot and semiquantitative RT-PCR, respectively. Furthermore, the same proteins were determined in vitro in U373, U251 and GaMG human glioblastoma cells using the same methods. HIF-1alpha protein regulation under hypoxia was also determined on mRNA level in vitro in GaMG and on protein level in U251, U373 and GaMG cells.

The results obtained show that there is no hypoxiainduced regulatory effect in the three glioblastoma cell lines studied in vivo or in vitro. In addition, no association between hypoxia and overexpression of GAPDH in human glioblastoma was observed. Conclusion: Therefore, we can conclude that the regulation of GAPDH mRNA and protein expression as a response to the hypoxic development in the tumor cell enviroment in vitro and in vivo is not an absolute phenomenon, but occurs as a cell-specific posttranscrip-tionally regulated event. Expression of GAPDH represents one of the alternatives of a housekeeping gene and can be used as a loading control in experiments with glioma cells. Therapeutic strategies for treatment of human astrocytic tumors involving GAPDH as target molecule do not represent a valid approach in conjunction with tumor hypoxia in the human brain. Gene expression studies related to cancer diagnosis and treatment are important. For this purpose, absolutely reliable housekeeping genes are essential to normalize cancer related gene expression. The most important characteristics of housekeeping genes are that they are present in all cells and that their expression levels remain relatively constant under different experimental conditions. However, no single housekeeping gene always manifests stable expression levels under all experimental conditions. Incorrect choice of housekeeping genes leads to interpretation errors of experimental results including evaluation and quantification of pathological gene expression. Methods: We examined (i) the degree of GAPDH expression regulation in human Human lung adenocarcinoma epithelial cell line (A549) and HT29, and HCT116 colon cancer cell lines, Hepatoma -1-6 mouse hepatoma and Hep3B and HepG2 mouse hepatocellular carcinoma cell lines under hypoxic conditions in -actin, serving asbvitro in comparison to other housekeeping genes like experimental loading controls, (ii) the potential use of GAPDH as a target for tumour therapeutic approaches was comparatively examined in vitro in on both protein and mRNA level, by Western blot and semi quantitative RT-PCR, respectively. Results: No hypoxia-induced regulatory effect on GAPDH expression was observed in the cell lines studied in vitro (Human lung adenocarcinoma epithelial cell line (A549), colon cancer cell lines HT-29, and HCT-116, Hepatoma -1-6 mouse hepatoma and Hep3B and HepG2 mouse hepatocellular carcinoma cell lines. Conclusions: As it is the case for mouse hepatocellular carcinoma, human colon cancer, mouse hepatoma and Human lung Adenocarcinoma, GAPDH represents an optimal choice of a housekeeping gene and/or loading control to determine the expression of hypoxia induced genes at least in glioblastoma. The results confirm the previous findings in human glioblastoma, that this gene is not an attractive target for tumour therapeutic approaches as well in lung cancer and colon cancer in Because of the lack of GAPDH regulation under hypoxia. Hypoxia, which is a crucial factor in tumour aggressiveness and its treatment resistance HIF-1a together with other factors is involved in NDRG1 regulation. NDRG1 expression analysis in gliobastoma in different hypoxic environments highlights some mechanism(s) involved in the regulation of this gene. Human U251, U373, GaMG & U87-MG glioma cells were exposed to (5-0.1%) O 2 , for 1 hour, 6 hour, 24 hour in addition to 24 hour with reoxygenation over 24 and 48 hour. Treatment with desferrioxamin (COCl 2 , 300 lM, 24 hour) served as a positive control, Incubation at (20% O 2 , 5% CO 2 ) as a normoxic negative control. HIF-1a expression were detected via western blots & mRNA (sq RT-PCR) while, NDRG1 was also detected via Northern blots. B-actin, b-Tubulin and 18 S RNA served as loading controls, respectively. HIF-1a nuclear extracts bound HRE element in a hypoxia dependent manner. With exemption of U87-MG (were NDRG1 was strongly expressed, independent of O 2 level, no NDRg1 (protein) expression was displayed in a normoxic environment in all other cell lines and at 5% O 2 . A substancial degree of NDRG1 expression, with a maximum at 0.1% and relative stability during reoxygenation emerged at (1-0.1%) O 2 . HIF-1a was moderately -strongly expressed after 1 hour 0.1% O 2 , and stable up to 48 hour reoxygenation after 24 hour at 0.1% O 2 . NDRG1 protein levels in human cancers aid in disease diagnosis. Glioma cells with minimal O 2 concentrations display a substancial NDRG1 expression with a relatively long stability. NDRG1 inhibition opens anti tumor therapeutic approaches potential. NDRG1 is regulated by the transcription factor hypoxia induced factor 1 -a (HIF-1a) and not by epidermal growth factor 1 (Egr-1).

omentum with ascites, without bone involvement, is extremely rare. We present an anusual case of a man, 75 years old, who referred to the oncological department with an increased serum PSA and a medical history prostate cancer following by prostatectomy, six years ago. The computerized tomography (CT) scan of abdomen and pelvis disclosed a massive ascites, as well as secondaries in the omentum and mesenteric fat. Furthermore, bone scintigraphy did not reveal any evidence of osseous metastases. The cytological diagnosis of ascitic fluid was positive for adenocarcinoma, probably derived from prostate. The clinicopathological findings are discussed. Objective: To assess the utility of having a cytopathologist during CT-guided FNA of deep body sites. Methods: From 2000 to 2008, 1550 FNA from 1525 patients were performed. The sampling was performed under CT guidance using a 18G needle. Smears were prepared by a cytopathologist, two slides were immediately stained with Giemsa; all FNA's were associated with on-site immediate microscopic evaluation, to assess adequacy and, when possible, a preliminary diagnosis was made. Cytohistological correlation was available in 255 cases obtained by subsequent surgical excision. Results: Cytology was negative in 416 (26.8%) cases, suspicious in 33 (2.1%), positive in 986 (63.7%) and inadequate in 115 (7.4%). Surgical specimen were negative in 40 (15.7%) cases, while 215 (84.3%) cases were positive for malignancy: these results were compared with corresponding cytologic reports. Sensitivity, specificity and accuracy of cytology were thus 98.1%, 83.3% and 95.3% respectively. Conclusions: This series study confirms the high diagnostic accuracy of on-site cytologic evaluation of deep body site lesions to reduce inadequacy rate. Cytologic sampling is considerably less expensive than biopsy techniques and it is associated with lower patient morbidity rate. Moreover, diagnosis is very quick, often contemporary to sampling, allowing physicians to provide diseasespecific intervention and reduce hospitalization time, with consistent financial saving. Objective: Malignant mesothelioma (MM) of the peritoneum (MPM) is a rare aggressive tumor but with an increasing incidence correlated with increased asbestos exposure. The diagnosis of malignant mesothelioma in serous effusions is a diagnostic challenge in cytology due to overlapping differential diagnostic features between mesothelial proliferations, MM and metastatic cancer. In our paper we describe and discuss the morphologic findings and immunocytologic profile of a case of MPM with respect to the latest improvements in the diagnostic approach of MPM. Case Report: The diagnostic workup of a 70 years old man with a 6 months history of abdominal pain, swelling and anorexia revealed the presence of ascitic fluid and peritoneal implants on CT scan. Cytologic examination of the ascitic fluid showed numerous clusters of neoplastic cells often with papillary configuration, morphologically compatible with adenocarcinoma. However, the immunocytological investigation of neoplastic cells showed positivity to Calretinin. The rest of the immunoprofile of the neoplastic cell population was: Positive markers CK 5.6, WT-1, HMBE-1, CK 7 and membranous staining for EMA and Negative markers B72.3, BerEP4, CK 20, CEA, TTF-1 and Leu M1. The cytologic diagnosis of MPM was confirmed by positive biopsy from the laparoscopy that followed and the patient received the appropriate therapeutic protocol. Clinical history revealed a possible correlation to asbestos exposure since the patient was a construction worker. Conclusion: The difficulty in diagnosing a MPM resides to the fact that the morphologic patterns displayed by MPM simulate more common epithelial malignancies and also that reactive mesothelium and MPM can mimic one another to a great extend. The use of immunocytochemistry with a panel of antibodies in relation to the clinical and radiologic findings can render a reliable diagnosis of MPM.

Objective: To examine whether BAL can help the differential diagnosis between asbestosis and IPF. Methods: Retrospective chart review of 31 patients diagnosed with asbestosis and 21 patients diagnosed with IPF. All patients had a BAL performed as part of their initial evaluation. BAL total cells and differential were counted and all specimens were examined for asbestos fibers. Statistical analysis was performed with the SPSS 14 statistical program. Results: Asbestos fibers were present in all patients with asbestosis and none of the IPF patients. IPF patients had significantly less lymphocytes (5.8 ± 3.73%) and significantly more eosinophils (10.85 ± 3.66%) compared to patients with asbestosis (13.41 ± 3.11%) and (6.83 ± 5.25%) respectively. No difference was found in neutrophils, macrophages and mast cells. Conclusion: BAL cell differential of patients with IPF is different from that of patients with asbestosis. This difference could be involved in the pathogenesis of IPF. Methods: Retrospective review of patient files. Sarcoidosis was diagnosed in 84 steroid-naive patients based on clinical-radiological findings, a CD4/CD8 > 3.5 and lymphocytic alveolitis in bronchoalveolar lavage (BAL). Differential count was determined on a stained preparation by MCG and Papanicolaou staining. Lymphocyte subsets in BAL were analysed by dual-color FACS. Patients were classified in chest x-ray stages. Results: Neutrophils and eosinophils were significantly increased in the BAL, from stage I to IV. Macrophages were significantly decreased from stage I to IV. The percentage of NK cells showed a significant increase between stages I and II (16.77 ± 2.25 to 18.76 ± 1.20%), I and III (16.77 ± 2.25 to 20.50 ± 1.67%) and I and IV (16.77 ± 2.25 to 20.4 ± 1.67%). Conclusions: Variations were observed in alveolar cells and lymphocyte subsets in different stages of pulmonary sarcoidosis. Cells exhibiting cytotoxic activity (NK) were increased with stage progression. These cells may be involved in the pathogenesis of the disease.

Objective: Pathological diagnosis of diffuse or focal lung diseases could be established on tissue samples obtained on fine needle aspiration biopsy (FNAB).

Aim: The evaluation of chest x-ray, CT-scan or ultrasound guided FNAB for diagnosis of lung diseases in period from 2006 to 2008. Method: Tissue samples obtained on FNAB were formalin fixed, paraffin embedded and hematoxylin-eosin stained. Immunohistochemistry was performed when microscopically finding indicated this procedure. Results: The results of 923 lung FNAB biopsies were reviewed to evaluate this investigation despite the age and gender of the patients. On 612 (66.31%) tissue samples diagnosis was representative. Final diagnosis of primary lung malignancy was established on 580 and secondary lung malignancy on 12 biopsies. The most frequently squamous cell carcinoma was diagnosed (250), followed by adenocarcinoma (164), small (106) and large (28) cell cacinomas and the other, rare (23) lung carcinomas. One malignant mesothelioma was also diagnosed. Malignant cells without precised typisation were diagnosed on two samples. Four benign tumors were diagnosed by FNAB (two hamartomas and two lipomas). Two benign mesenchimal and five metastatic mesenchimal tumors were diagnosed by FNAB. Granulomatous disease considering tuberculosis was diagnosed in 10 patients. Lung inflammation was diagnosed on two tissue samples. Among all of them immunohistochemistry was performed on 80 lung biopsies for precised typisation of tumor type. Conclusion: It is concluded that FNAB usefulness is predominant in diagnosis of malignant primary and secondary lung tumors. Benign pulmonary focal tumors and diseases or diffuse diseases could be diagnosed in rare cases. Paraffin sections of FNAB biopsies could be successfully used for immunophenotypisation of lung tumors.

Objective: Local mediastinal tumors are deeply localized and pathological diagnosis could be established obtained on fine needle aspiration biopsy (FNAB). Aim: The evaluation of chest x-ray, CT-scan or ultrasound guided FNAB for diagnosis of deeply sited mediastinal tumors in the last 3 years. Method: Tissue samples obtained on FNAB were routinely formalin fixed, paraffin embedded and hematoxylin-eosin stained. Immunohistochemistry was performed when microscopically finding indicated this procedure. Results: The results of 200 mediastinal FNAB biopsies were reviewed to evaluate this investigation despite the age and gender of the patients. On 110 (55.00%) tissue samples diagnosis was representative. Most predominantly Hodgkin and non-Hodgkin lymphomas were diagnosed on FNAB (34). The majority of them were Hodgkin lymphoma (25). Thymoma, as anterior mediastinal tumor mass was diagnosed on 10 aspiration biopsies. It was noted that granulomatous diseases, tuberculosis and sarcoidosis, presented as mediastinal lymphadenopathy were diagnosed in nine patients. Metastatic carcinomas were diagnosed in 49 patients, most of them were adenocarcinomas of different, predominantly lung origin. Small cell lung carcinoma (15) was diagnosed in cases with notable mediastinal lymphadenopathy. Metastatic squamous cell lung carcinoma (7) and one malignant melanoma were also diagnosed. Among mesenchimal tumors (4), neurilemmoma was diagnosed twice. Mediastinal teratoma and plasmocytoma were diagnosed once, each. Inflammation was detected once. Among all of them immunohistochemistry was performed on 70 mediastinal biopsies for precised typisation of tumor type, mostly lymphomas, thymomas and metastatic carcinomas. Conclusion: FNAB usefulness is predominant in diagnosis of Hodgkin and non-Hodgkin lymphomas and thymomas, respectively. Diseseses with mediastinal lymphadenopathy as a leading symptom, sarcoidosis and small cell lung carcinoma, could be diagnoses by FNAB in high percentage. Paraffin sections of FNAB biopsies are useful in immunohistochemical procedures and precised diagnosis of mediastinal lymphomas as well as thymomas and mediastinal carcinomas.

Introduction: Bladder CIS is a high grade non-invasive urothelial neoplasia with an elevated rate (> 50%) of progression to invasive carcinoma. Unlike other superficial or early bladder tumors, the clinical diagnosis by cystoscopy is difficult. Objectives: To analyze the diagnostic value of voided urine cytology specimens in carcinoma 'in situ' (CIS) of the bladder in liquid based (LBC) and convencional cytology. Materials and Methods: Histologic and cytologic samples from patients with bladder CIS diagnosed by histology in our archives from the past seven years, were retrospectively reviewed. Inclusion criteria for the study were: (i) histological sample with CIS and, at least, one voided urine specimen from less than a year before diagnosis, and (ii) no urinary tract malignancy other than CIS, either simultaneous or within the previous 12 months, and (iii) at least 2, 5 year total clinical follow up after the diagnosis of a previous bladder tumor other than CIS. Results: There were 45 cases meeting the inclusió n criteria. In 31 cases, the samples were processed by LBC. The remaining 14 patients had samples with CC, by the millipore filter technique. In 23 patients, there was no history of bladder tumor other than CIS, with a follow up ranging from 6 to 72 months. The remaining 22 cases had a 26 to 126-month follow up. Thirty three (73.3%) patients had at least one positive previous diagnosis by cytology. There was no statistically significant difference between LBC or CC-processed specimens in predicting malignancy. The quality of the sample was considerably better in LBC when compared to CC specimens. Conclusions: (i) cytological analysis of voided urine specimens has a high diagnostic rate in bladder CIS, (ii) liquid-based cytology yields high quality samples with similar results to CC in the diagnosis of CIS. (iii) Unlike with most early bladder neoplasias, urine cytology is a very useful diagnostic tool for bladder CIS follow up.

M. Albuquerque* , † , P. Mendonç a*, L. Antunes* , † and A. Ribeiro † *Escola Superior de Tecnologia da Saude de Lisboa, Instituto Polite´cnico de Lisboa, Portugal, CHLN -Hospital Pulido Valente, Lisboa, Portugal

Objective: To describe the Liesegang rings, that are laminated structures well known in the chemical field, but rarely recognised 'in vivo', appearing in association with inflammatory or cystic procedures. Materials and Methods: Left pleural effusion, of serohematic aspect, belonging to a 58 year old male, smoker, with a clinical diagnosis of a probable pneumonia. Pleural fluid received in cytolyt, processed, and fixed in preservcyt. The liquid base material was processed in the THINPREP 2000 Ò . Results: The pleural fluid presented round/oval laminated eosinophilia structures with parallel concentric bands in a background of some neutrophils, lymphocytes and histiocytes. The pleural biopsy revealed a fibrous conjunctive 'mixoide' tissue with fibroblast type cells and inflammatory infiltrate with a predominance of polymorphonuclear and proliferation of small vessels. Conclusions: The cytology diagnosis was negative for neoplastic cells and the structures were identified as Liesegang rings. Objective: Urinary tract cytology is the most important diagnostic tool for bladder cancer screening. During a national quality audit within our laboratory the conventional cytospin urinary tract samples were criticized for low quality. For some years thin layer technique is available in our department and has been applied successfully in cervical and pulmonary cytology. Furthermore, Thinprep cytology offers the opportunity for additional tests like in situ hybridisation. Therefore conventional cytospin preparations were compared to the Thinprep technique in urinary cytology. Materials and Methods: 103 urinary tract samples were split equally and processed by both Thinprep and cytospin method. Quality parameters (cellularity, background, cell preservation and nuclear detail) were compared and scored. Also diagnostic efficacy was evaluated. Results: Thinprep specimens scored equal or better on all qualitative parameters. Especially background artefact, cell preservation and nuclear details scored better, in respectively 52, 63 and 43% of cases. Cellularity scored equally in 55% and better in 27%. There were only minimal diagnostic differences. In both techniques 84% was benign and 7% malignant. 4% showed atypical cells, indeterminate for malignancy and 5% was rejected because of low cellularity. One sample was diagnosed as low grade transitional cell carcinoma in the conventional specimen and high grade in Thinprep. Histologically high grade transitional cell carcinoma was confirmed. Conclusion: Thinprep urinary tract samples showed cleaner background, better nuclear details and better cell preservation. However, despite better quality, Thinprep technique did not show major diagnostic improvement, as compared to conventional method. Because of the additional cost of Thinprep, we recommend to analyse a cytospin urine specimen first, and store the remaining urine in a liquid based preservation fluid. When necessary, a second thinlayer specimen can be subjected to additional tests. These way good diagnostic facilities are used at relatively low cost. Objective: Urine cytology is hampered by its low sensitivity for malignancy when traditional preparation methods are used. Therefore, we compared the diagnostic performances and effects on diag-nostic criteria in CytoRichRed (CRR) (AXLAB, Denmark) fixed Surepath preparations and conventional slides from a series of urine samples. Materials and Methods: Urinary samples from 196 patients (160 voided urine, 18 bladder urine and 25 bladder washings, in all 203 samples) were split, one part was fixed in CRR and prepared manually according to the Surepath protocol, and the rest was prepared according to the routine of the department (Poretics filter) The filter and the Surepath slides were evaluated in a blinded fashion, the diagnoses were compared, and related to histology, if available. Results: Surepath and filter diagnoses disagreed in 34/196 cases. In nine cases the discrepancy was due to insufficiency of the filter preparations. There were more benign diagnoses in Surepath than in filter preparations (159 versus 140) and fewer atypias (18 versus 24), but the numbers of malignant diagnoses were similar (23 versus 21). In 5/6 histologically diagnosed cases the Surepath and filter diagnoses agreed. One carcinoma was diagnosed as atypical with Surepath and benign with the filter method. The Surepath slides were usually easier to evaluate due to the removal of blood and salt crystals, but in bladder washings decreased cell cohesion and lack of preservation of poorly differentiated cancer cells were seen. Conclusions: CRR fixation and Surepath preparation of urine samples reduced the number of insufficient or atypical cases, but the number of malignant diagnoses was similar with the two methods. Decreased cell cohesion and lack of preservation of poorly differentiated cancer cells, seen in bladder washings, was a potential pitfall.

Objective: Rapid evaluation has been proposed as a method to improve the yield of guided FNAB. The aim of our study is to assess the value of immediate evaluation of respiratory specimens. Materials and Methods: 370 consecutive adult patients underwent guided (CT, X-rays, ultrasound) FNAB of lung and transbronchial needle aspirations of mediastinal lymph nodes. In the bronchoscopic and radiology department, direct smears were prepared from aspirated material. Smears were immediatly wet-fixed in Delaunay fixative, quickly stained by Toluidine Blue and covered. Cytopathologist or trained cytotechnologist assessed the specimen adequacy by telecytology system (Coolscope II, Nikon, Japan). The assessment included celullarity, preservation of cells and the presence of diagnostic material. Specimens were assessed as adequate or inadequate. Results were compared to final diagnoses (nonrepresentative, negative, positive). Results: Out of 370 specimens, 234 specimens (63.2%) were diagnosed without rapid assessment. The percentage of nonrepresentative samples whithout rapid evaluation is 41.9% (98/234). From 136 specimens (36.8%) which underwent rapid evaluation, 44 (32.4%) were inadequate and 92 (67.6%) adequate. Only 22.1% (30/136) of final diagnoses were nonrepresentative, including samples which were assessed in rapid evaluation as inadequate. In three patients (3/30) , where the obtained material was recognized as adequate, final diagnoses were nonrepresentative. Objective: The evaluation of thyroid nodules using the split-sample processing in US FNA specimens and the comparison of liquidbased method with the conventional one. Patients and Methods: In all, 806 thyroid US/FNAs from 699 patients, admitted to the 2nd Department of Radiology of our hospital, were included in this study. Two air-dried May-Grü nwald-Giemsa (MGG) stained slides and one slide prepared with the ThinPrep Ò technique (Cytyc Co., Boxborough, MA, USA) were examined for diagnostic purposes. Results and Conclusions: Cytological diagnosis concerned 740 benign lesions, 34 suspicious and 32 neoplasms. In 112 cases, cytological diagnosis was made by the ThinPrep Ò slide alone, while in 14 cases by the MGG smear alone (x2 = 20,22, P < 0,001). Five cases of neoplasms (three papillary carcinomas, one Hü rthle cell tumour and one follicular tumour) were detected by the ThinPrep Ò technique alone, while two papillary carcinomas by the MGG smear alone. Although no statistically significant difference was found between the above studied methods, as it concerns the diagnosis of neoplasms, liquid-based method alone may provide an effective tool for preparing thyroid US/FNA specimens, offering a better quality of smears, a cytotechnical convenience in US Outpatient's Department and the advantage of further ancillary techniques in the same specimen, in which cytomorphological diagnosis was made. Objective: Three markers -Galectin-3 (GAL3), Ki-67 e cytokeratin-19 (CK19) -were availed to define their respective accuracy and possibility of association to aim better diagnosis precision in the differentiation of follicular benign and malignant thyroid nodules. Methods: Retrospective study of anatomic pathology and cytological reports of follicular thyroid nodules in 34 patients submitted to fine needle aspiration (FNA) guide by ultrasound that was analyzed by the 'Center of Pathology of Curitiba -Paraná ', between January 2003 and July 2008. The material was analysed by immunocytochemistry (ICC) and was correlated to the histopathologic diagnosis. Results: The best performance of all statistics's criteria correlation occurs when it is considered malignant suggestive nodules to the ICC with the minimum of two positives markers, with 100% sensibility, 75% specificity and 93.3% accuracy results. Conclusion: The present study conclude that in thyroid lesions with follicular cytology, ICC analysis may be used in identification of malignant lesions when utilized a combined panel with GAL3, Ki-67 and CK19 and at least two of them markers were positives. Aim: To determine if FNA cytology is useful in the diagnosis of metastatic germ cell tumors. Materials and Methods: A FNA was performed on 66 surgical specimens (61 testicular specimens, 2 pulmonary nodules, 1 retroperitoneal mass, 1 thoracic-abdominal mass and 1 inter aortic-cava node). Aspirates were either fixed in 95% ethanol or air dried and stained with Papanicolaou or Diff-Quick methods, respectively. Two independent observers evaluated the cytologyc material. Concomitantly the histologyc diagnostic was made. Results: Twenty six seminomas were diagnosed by histology. The 40 remaining cases were non seminomatous germ cell tumors (GCT): 23 Mixed GCT, one Yolk Sac Tumor, six Embryonal Carcinoma, two Choriocarcinomas, six Teratomas and two Leydig tumors. The material obtained in four cases was insufficient to make the diagnosis (two seminomas, one embryonal carcinoma and one mixed GCT). The cytologic findings -tigroid background (47.8%), lymphocytes and plasma cells (100%), large isolated cells and/or loose sheets (100%), round nucleus with conspicuous nucleoli and fine granular chromatin (100%), stripped nuclei and chromatin threads (43.4%)-enabled a diagnosis of seminoma in all the smears. In non seminomatous GCT other different features were observed, namely: three-dimensional clusters of cells, papillary groups, marked pleomorfism and marked necrotic or mucinous background. Conclusion: The cytologic findings observed in testicular GCT are useful to differentiate between seminomatous and non seminomatous GCT and maight be extrapolated in metastatic GCT. Objective: We determined the anticancer activity, both in vitro and in vivo, of phosphoaspirin, 3-((diethoxyphosphoryloxy) methyl) phenyl 2-acetoxybenzoate, a novel derivative of aspirin. Materials and Methods: Phosphoaspirin was synthesized by linking a phosphate group (diethyl phosphate) via a spacer moiety to the conventional aspirin. HT-29 colon adenocarcinoma cells from ATCC, were counted using the MTT assay and proliferation and apoptosis was determined. HT-29 cells were injected subcutaneously into the flank of nude mice. Phosphoaspirin, 100 mg/kg body weight, was injected intraperitoneally daily for 21 days. The size of the xenographs was recorded and at the end of the experiment the tumors were excised and weighted. Proliferation and apoptosis of cancer cells was determined histochemically by staining for proliferating cell nuclear antigen (PCNA) and by terminal deoxynucleotidyl transferase-mediated deoxy-uridine triphosphate-biotin nick end-labeling (TUNEL) assay respectively. We calculated the proliferation index and the apoptotic index, using a light microscope at magnification · 400. Data was analyzed using t-test. Results: Phosphoaspirin inhibits the growth of HT-29 cells. At its IC50 concentration at 24 hour (276.6 lM), phosphoaspirin inhibited proliferation by 19% compared to control, whereas the apoptotic cells were 38% of the total, compared to 4% in untreated controls. Phosphoaspirin also inhibits the growth of HT-29 xenotransplants. The weight of the tumors at sacrifice showed a 57% reduction, while proliferation of cancer cells was reduced by 18.13% compared to control and apoptotic index showed an increase of 94.6%. Conclusion: Our studies demonstrate a strong cytokinetic effect of phosphoaspirin, a finding that needs further investigation for the role of phosphoaspirin in cancer control. Background: Historically, the diagnosis and monitoring of multiple myeloma (MM) and plasmacytoma has been based on bone marrow (BM) aspirations and surgical biopsies. FNA has been used to diagnose primary and recurrent MM and plasmacytoma, but its role in this application has not been well reported. The aim of this study was to determine the role of FNA in the diagnosis, follow-up, and treatment of patients with MM and plasmacytoma. Design: A computerized search of the cytology laboratory information system was performed and FNA cases diagnosed as MM or plasmacytoma were identified over an 18-year period (May 1990 through April 2008). All correlating hematopathology and surgical pathology reports and medical records were reviewed, as were selected slides.

Results: A total of 70 FNA specimens from 68 patients were included in the study. Of these, 47 were diagnosed as MM, 14 as plasmacytoma, eight as plasmacytoma/myeloma, and one as malignant plasma cell lesion. Patients ranged in age from 37 to 86 years (mean age: 64) and included 44 males (65%) and 24 females (35%). The sites of the FNAs were soft tissue/lymph node (40 cases, 57%), bone (27 cases, 39%) and viscera (three cases, 4%). In 42 cases (60%), FNA was performed on patients with prior histologic diagnoses of MM or plasmacytoma. The time interval between the primary diagnosis and the FNA ranged from 3 days to 23 years (mean: 37 months). In two cases, the date of primary diagnosis was unknown. Concurrent histopathologic/bone marrow (HP/BM) diagnoses were made in six cases (9%). In 22 cases (31%) there was no known previous or concurrent HP/BM diagnosis. A subsequent confirmatory pathologic diagnosis was obtained in 33 cases (47%) while 37 FNA cases (52%) had no confirmatory diagnosis. In three of eight FNA cases with a subsequent HP/BM diagnosis, treatment was initiated before confirmation was obtained. Of 14 FNA cases (20%) lacking any previous, concurrent, or subsequent HP/BM diagnoses, five patients (36%) received treatment based on the FNA, 2 (14%) received no treatment, and 7 (50%) had no available clinical follow-up information. Flow cytometry was used as a diagnostic aid in 18 cases (26%) and immunostains were performed in 14 cases (20%). In seven cases (10%) both flow cytometry and immunostaining were performed. Conclusion: FNA is a useful, rapid, relatively inexpensive method for diagnosing both primary and recurrent MM and plasmacytoma. It should be considered the diagnostic test of first choice for diagnosing disease recurrence and, at our institution, has been used as the sole basis for initiating therapy in some patients. Objective: Assessment of HER2 status is mandatory for treatment of patients with invasive breast cancer. According to recommendations (ASCO/CAP 2007) HER2 should be assessed on histological specimens with a validated immunohistochemical or FISH method. However, in some patients histological material is not available and cytological specimens could be an alternative. The aim of our study was to optimise an imunocytochemical (ICC) protocol for HER2 assessment on cytological material. Materials and Methods: HER2 protein was detected using c-erB-2 Oncoprotein antibody (DAKO; A0485). The results of ICC reactions were evaluated according to the histological guidelines. ICC protocol for HER 2 assessment was optimised on cytospines made from cultured cell lines (SK-BR-3, MDA-MB-453, MDA-MB-175 and MDA-MB-231) with known levels of HER2 expression (pos 3+, moderate 2+, neg 1+ and neg 0). The protocol was tested on 36 cytological specimens of primary or metastatic breast carcinomas. Results of ICC reaction were compared to results of a routine FISH method (PathVysion HER2 DNA probe kit, Abbott Molecular Inc) performed on cytospins from the same tumors which were used for ICC method.

Results: In the four cell lines the optimised ICC protocol resulted in a membrane stain that corresponded to the declared level of protein expression. Strong HER2 protein expression (3+) obtained by ICC was found in 6 out of 36 cases (17%), moderate (2+) in 3 (8%) and negative (0 or 1+) in 27 (75%). This outcome was confirmed also by Fish: HER2 gene was amplified only in six cases (17%) which were ICC strongly (3+) positive. Conclusion: HER2 can be reliably determined on cytological samples of primary or metastatic breast carcinomas using ICC method. Objective: We report a rare distinctive variant of invasive breast carcinoma characterized by sebaceous differentiation of tumor cells as a primary breast carcinoma. Our report covers the first case of sebaceous carcinoma recorded in Croatian medical literature. A search of the literature revealed a total of just a few cases reported. Materials and Methods: 85-year-old female presented to our hospital year with a two months history of swelling of the left breast under the mamila. Segmentectomy was performed after mammograhy and US guided FNA. Results: Cytology diagnosis was: suspicious sebaceous or lipid rich carcinoma but we also considered an apocrine carcinoma.The aspirates showed a lot of clusters and single, mostly uniform cells with visible clear cytoplasmatic vacualization and a coarse cromatin structure. A gross examination revealed a tumor measuring 2.5 cm with its portion reaching the excision margin. Hystologically, the tumor was made up of loose connective tissue stroma with solid clusters of large cells of markedly light-colored cytoplasm and areas of squamous morulas. Histology could not distinguish whether this was a primary sebaceal cancer of the breast or of the skin. Immunohistochemistry showed negative reaction for ER, PR and HER2/neu. After 10 months the patient came with a new swelling under the postoperative scar. The cytology picture of the smears was the same as previous one and mastectomy was performed.Tumor was 5 cm in diametar and hystologically was the same as the previous one. The lesion had no connection to the overlying skin or the nipple. Immunohistochemistry showed the tumor cells positivity to cytokeratin and a negative reaction for GCDFP-15, ER, PR and HER2/neu. Conclusion: Sebaceous carcinoma of the breast is extremely rare.The origin of sebaceous cells in breast carcinoma and their biological behavior is still unclear. Objective: We report a case of haemangiopericytoma of the parotid gland in a 29-year old woman. Haemangiopericytoma is a rare soft tissue tumor of vascular origin, with unpredictable clinical behaviour. A location in parotid gland is exeptionally rare (a search of the world literature revealed only 30 cases reported). Materials and Methods: A 29-year old woman presented with a 2-year history of the left parotid gland swelling.Total parotidectomy was performed after a clinical and Ultrasound examination, and US guided FNA. Results: The aspirates were moderately cellular with bloody background, and were comprised of cohesive fragments of cells with oval to spindle shaped nuclei and finely granular chromatin showing mild pleomorfism. Naked nuclei were also seen. The cytologic diagnosis was: suspicious mesenchymal tumor. After operation, gross examination revealed salivary gland tumor measuring 5 : 4 : 2 cm. On section a well-circumscribed pinkgreyish, firm tumor, measuring 4 : 2 : 1.5 cm was found. Microscopically, the tumor was rich in small wessels lined by single layer of endothelial cells surrounded by tightly packed round to oval spindle cells with indictinct cell borders. Perivascular hyalinisation and myxoid changes were focally found. Mitotic figures were rare (1/10 hpf). Immunohystochemmicaly the tumor cells were positive for CD 34 and Vimentin and negative for Cytokeratin and SMA. A final diagnosis of haemangiopericytoma of the parotid gland was made. Conclusion: This is a first case of haemangiopericytoma of the parotid gland recorded in Croatian literature. Because of its rarity, we wanted to present it as a case. Objective: Endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) is a minimally invasive technique, increasingly used for the assessment of digestive organs. The technique is also important to provide a primary diagnosis of lung cancer, as well as mediastinal staging. Endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-FNA) is a complementary technique that allows sampling of lymphnodes and masses in the middle mediastinum. These techniques are complementary and achieve a complete medical mediastinoscopy, knowing that an optimal management of patients with lung cancer requires accurate cell typing of tumours and staging at the time of diagnosis. We performed a preliminary study including 30 patients with clinical suspicion of malignant tumours, from which 25 patients were assessed by EUS-FNA within the tumour and five by EBUS-TBNA within the tumour and in the lymph nodes. For establishing the tumoral cells type, we performed immunocytochemical studies with several antibodies for squamous cell carcinoma and adenocarcinoma. None of the cases haven't the morphology suspicious for small cell lung carcinoma on H&E stain. Materials and Methods: The patients have been examined by EUS at the Research Center in Gastroenterology and Hepatology, University of Medicine and Pharmacy Craiova. EUS-FNA within the tumour combined with EUS-FNA within the aorthopulmonary window lymphadenopathy was performed in 25 patients, while the other five patients were assessed by EBUS-TBNA within the tumour and in the lymph nodes. We obtained two types of specimens: smears, at least 10 smears per case, which were prepared directly from the aspirate, and cell blocks from wax embedding of the material after centrifugation. The sections from cell blocks were subsequently stained with Haematoxylin -Eosin and with special immunostains in case of the malignant tumours. The smears have been stained either by Giemsa stain or Papanicolaou stain. Immunocytochemistry was performed on cellular blocks obtained from the tumor and we used the following antibodies for establishing the immunophenotype of the tumor cells: anti-CK 7, anti-cytokeratines AE1/AE3, anti-34bE12, anti-TTF1, anti-CEA, anti-chromogranin, anti-synaptophysine. For the evaluation of the proliferation rate of the malignant tumours we assessed the Ki67 proliferation index (Ki-67 LI). Results: Of the 25 patients assessed by EUS-FNA, 20 had a positive FNA, confirming the diagnosis of malignancy, and in four patients the result was negative. One of the patients could not be assessed by EUS-FNA because of the impossibility of finding a suitable puncture route, without interfering with the main vessels (the aorta, the pulmonary artery). All of the five patients assessed by EBUS-TBNA had positive results for malignancy either on smears or cell blocks. On immunocytochemistry, malignant tumor cells expressed cytokeratines AE1/AE3 and 34bE12 in 19 cases, consisting mostly of squamous pulmonary tumours. Immunostains positive for CK 7, TTF 1 and CEA in six cases was consistent mostly with adenocarcinoma. As expected, the Ki67-LI was higher in tumor cells with severe nuclear atypia. Conclusions: In conclusion, EUS is highly sensitive for diagnosing lung cancer, while FNA improves not only the specificity, but also the accuracy. In addition, EUS-FNA is an excellent minimal invasive technique, especially when the standard techniques of histological diagnosis are negative, influencing the therapeutic actions and preventing unnecessary invasive techniques such as: toracotomy, toracoscopy and mediastinoscopy. In association with EBUS-TBNA it can sample lymph nodes and masses in the middle mediastinum. The immunocytochemical examination on cell blocks can establish the immunophenotype of the tumour in the cases where other imaging methods could not obtain a suitable biopsy. Objective: The aim of this study is to evaluate the usefulness of the fine needle aspiration (FNA) and the ability of the Cytopatholo-gist to provide accurate diagnosis for the study of superficial and deep-seated space-occupying thyroid's gland lesions. Materials and Methods: We present ultrasound-guided (US) FNA specimens from 980 patients, 420 males and 560 females aged from 15 to 90 years (mean: 49.5 years) with thyroid lesions, in the last 3 years.The sampling was performed by the Cytopathologist under U/S guidance. The smears were rapidly stained using a quick method (Diff-Quick) mainly in order to examine sampling adequacy. Results: Satisfactory diagnostic material was obtained in 950 of the 980 FNA's (96.9%). All cases were diagnosed cytologically and 350 FNA's (35.7%) confirmed histologically with the following diagnoses: Nodular or hyperplastic goiter 256 cases (73.10%) and thyroiditis 26 cases (7.42%) (four subacute, one acute and 21 chronic lymphocytic thyroiditis of Hashimoto). Thyroid neoplasms were found in 68 cases (19.42%) of which follicular adenomas six cases (1.71%), papillary carcinoma 48 cases (13.7%), Hurthle cell tumor six cases (1.71%), medullary carcinoma six cases (1.71%), anaplastic carcinoma one case (0.28%) and metastatic carcinoma one case (0.28%). Ten cases with hyperplastic goiter and chronic lymphocytic thyroiditis have also a microscopic focus of papillary carcinoma m.d. 4 mm. For the evaluation of thyroid lesions, adequacy of the cytological sample is required as well as accurate evaluation of the cytomorphological features. Conclusion: FNA can be a safe, fast and accurate method which provides rapid and valid results in the diagnosis of the thyroid lesions. The attentive study of the cytomorphological features prevents mistakes and provides useful information to the clinicians for further follow-up and treatment of patients with thyroid lesions. Objective: Ultrasound guided thyroid FNA is performed by physicians with various backgrounds, with variable amounts of formal training and clinical experience, being utilized for not only previously unsuccessful thyroid FNA procedures, but also for small non-palpable nodules (< 1 cm), with an improvement in sample adequacy and diagnosis accuracy. However, different specialists have distinct competencies and a multidisciplinary clinical team performing FNA would be a perfect scenario for unison of these competencies and optimal results. Our present work compares the results of ultrasound guided FNA smears obtained from a joint multidisciplinary clinical team approach (radiologist, pathologist and head and neck surgeon) with those performed solely by a radiologist and non-ultrasound guided FNA performed by a pathologist alone. Materials and Methods: FNA performed by the three groups (clinical joint, radiologist and pathologist) in our service in the last five months were reviewed. Samples were assigned as satisfactory or non-satisfactory. The satisfactory results were also compared according to the diagnosis provided, using chi-square and fisher exact tests. Results: The multidisciplinary approach showed a smaller amount (6.7%) of non-satisfactory samples (P = 0.036) as compared to the other groups (17.9% -pathologist only; 24.0% -radiologist only). The rate of malignant diagnosis was higher (P = 0.023) using this method (13.3% versus 4.3% from the other groups combined). Conclusion: The presence of a clinical joint team in the FNA room improved the rate of adequacy and diagnosis accuracy. Whenever possible, this approach should be utilized for decreasing complications and permitting a mutual interchange for assessing the thyroid nodule, reaching for a better result. Objective: Rhino-orbital-cerebral mucormycosis (ROCM) is an uncommon and aggressive fungal infection occurring in several immunocompromised states, including diabetes, the most common predisposing factor. The diagnosis of ROCM is usually based on direct microscopy of fluid from the nasal/sinus mucosae or through histopathology. The finding of septate hyphae with right angled branching is determining for the diagnosis of the fungus. We present here a case of a 41 year old diabetic male in which the diagnosis was assembled through cytological examination of the nasal smear. Report: 41 year old male, with recent diagnosis of Diabetes and Hipertension, under no treatment. At the admission, patient presented periorbital erithema, fever and dizziness, Kusmaul breathing, tachycardia, dehydration and ketonic breath, with periorbital celulitis and ptsosis on the right side. CT scan showed a big tumor on the right brain lobe and edema. Patient progressed to coma, and underwent a MRI suggesting angioinvasive fungal infection. At this time, the nasal smear was sent to examination, showing septated hyphae, being diagnostic of mucormycosis, later confirmed through culture, with Rhizopus sp. growth. After the cytological diagnosis, antifungal treatment was introduced, with a progressive improvement of the patient condition. Conclusions: ROCM must be included as a differential diagnosis for brain tumors in imunnocompromised patients, especially diabetics, and the use of citology is an effective and inexpensive way to provide fast and precise diagnosis in theses cases, permitting a proper choice of handling and treatment. Cryptococcosis occurs through the world and is caused by a fungus Criptococcus neoformans. Clinical disease usually develops in immunocompromised hosts, particularly those with AIDS. Diagnosis could be done in cytology by identification of fungus using special stains.-Fine needle aspiration (FNA) cytology is a technique minimally invasive, well tolerated by patients, safe and inexpensive. We describe three cases of cryptococcosis in Mozambican patients, diagnosed by FNA cytology. Sites of involvement included lymph nodes, face and mediastinum, respectively. The first two patients were HIV positive. In all cases was possible the identification of round fungal structures with a clear halo in the periphery (capsule), confirmed by PAS, Grocott and Masson Fontana stains.Our presentation shows the clinical importance of FNA cytology for the diagnosis of infectious diseases, namely cryptococcosis, in low resource settings, were the burden of this type of pathology is high, particularly after the advent of HIV infection. Object: The paracoccidioidomycosis is a deep mycotic infection, caused by Paracoccidioides brasilienses. The fungus can be founded in Central America and South America countries. It's endemic in some areas in Brazil. The fungal reach the human host by inhalation and from lungs it can disseminate to several organs. The most of the cases present initially as a pulmonary infection. The most evident manifestation in the buccal cavity is as ulcerated lesion, or as area mulberry-like that can be mistaken with neoplasie. The objective of this study is to show that cytologic examination can be use to diagnosis this pathology. Materials and Methods: Five patients were examined; four were submitted to exfoliative smears and one to fine needle aspiration (FNA). They present extensive buccal lesions with ulceration and infiltrative features. Two lesions had infiltrative features similar to neoplasie but in anothers the clinical appearance was a paracoccidioidomicosis. One patient presented the salivary gland enlarged. The smears were transferred dry glass slides, fixed with polyetylenoglycol and processed for cytogical assessment using the Papanicolaou, PAS and Grocott staining procedure. The cytologic diagnosis of paracoccidioidomicosis was confirmed when the fungal cells with classic morphology (round or oval cell with 5 to 15 microns in diameter, reproducing by single or multiple budding) was found. Results: The fungal cells were present in tree cases (two in cytologics exfoliated smears and one in FNA) confirming the cytologic diagnosis of paracoccidioidomicosis. The patients were treated and followed periodically with cytologic examination without alterations. In the other two cases, only inflammatory alterations was found, the cytological smear was repeated without alterations and the patients had been followed. Conclusion: The cytologic examination obtained by the exfoliative smear or through the FNAF is an efficient tool in the diagnosis of paracoccidioidomicosis. If the test is positive the patient should be treated and in case of negative results, he can be followed with aditional cytologic examination or submitted to biopsy. Object: The oral cavity epithelium suffer changes in most of inflammatory conditions, witch make possible histological diagnosis. Frequently, it is necessary to correlate the histological alterations that take place in the epithelial lining with the one at the chorium, to make a correct diagnosis. Some of these pathologies are susceptible for malignant transformation, needing a biopsy for their detection. The objective of this work is to use cytology for diagnosis and check the method's efficiency for detecting possible malignant transformation. Materials and Methods: Ninety-five smears were obtained from patients with clinical diagnosis of trauma, lichenoid reaction, lichen, abscess, erythroplakias, leukoplakias, aphthous stomatitis, collagenous diseases, congenital epulis, nicotinic stomatitis, pemphigus vulgaris, herpetic stomatitis and other inflammatory conditions. The smears were processed for cytological assessment using the Papanicolaou staining procedure. Results: Inflammatory alterations were observed in all the examined cases. The presence of anucleated cells were found in 54% of the leukoplastic lesions indicating hiperkeratosis. One case of atypical cells of undetermined significance (ASCUS) was found in a clinical suspicion of lichen, being directed to biopsy. All the cases with clinical diagnosis of pemphigus presented cytological atypias, and they were described as atypical cells of undetermined significance (ASCUS), compatible with pemphigus. In two of these cases, Tzanck typical cells were observed. The presence of reactive nuclear alterations in cytological examination of two lesions caused by trauma, one of erythroplakias, two of leukoplakias, and one of lichen indicate cytological follow up. Conclusion: Nonspecific inflammatory alterations dominate the cytologic findings in oral mucosa smears. Intense hyperkeratosis can mask a malignant epithelial injury. Even though, cytological examination is a useful tool to detect neoplastic alterations in epithelial mucosa. The finding of Tzank cells in smears allows a conclusive diagnosis of oral pemphigus.

P. Liodantonaki*, G. Apostolou*, K. Kyriakou*, G. Bairaktari*, N. Arnogiannaki † and D. Daskalopoulou* *Cytology Department, Saint Savvas Oncologic Anticancer Institute, Athens, Greece, Pathology Department, Saint Savvas Oncologic Anticancer Institute, Athens, Greece Aim: To indicate the role of intraoperative cytologic examination in the diagnosis of orbit tumors that are uncommonly encountered. Materials and Methods: In a period of 10 years 321 cases of primary and metastatic orbital tumors were diagnosed cytologically in our department. Nineteen of the above cases were selected for this study.The selection criteria were the uncommon location of the tumors and the special interest of their cytomorphologic features. Cytologic imprints were obtained after surgical excision of the tumors.The specimens were stained with a quick Giemsa stain for a rapid intraoperative diagnosis and subsequently with Papanicolaou stain as well. Results: The cytologic examination revealed seven meningiomas, four lymphomas (three NHL high-grade, one NHL high-grade MALT), two myelomas, two primary melanomas, one lacrimal gland glandular carcinoma, one adenoid cystic carcinoma metastatic from salivary gland and two peripheral nerve neoplasms (one neurilemmoma and one potentially malignant neoplasm).Correlation with the histologic diagnoses revealed that in 18 out of the 19 cases the cytologic and histologic diagnosis were in concordance. Only in the case of the cytologically diagnosed potentially malignant peripheral nerve neoplasm, there was discrepancy, the histologic diagnosis being squamous cell carcinoma. Conclusion: Our study demonstrates that intraoperative cytology is a simple, quick and accurate method that can contribute significantly to the diagnosis of uncommon orbital tumors.

sarcomas is 97%. The assigned cytologic grade accurately reflects the histologic grade in 90% of sarcomas when segregated into high and low grades. Specific FNAB diagnoses (subtyping) are correct in 14% to 54% of the cases in combination with ancillary studies, clinical and radiologic findings. Conclusions: The FNAB of soft tissue mass lesions is used as an accurate and minimally invasive method for the initial cytopathologic diagnosis of the primary soft tissue tumors. Nevertheless, the accuracy and therapeutic significance of FNAB for histologic subtyping of soft tissue tumors are still controversial. Objective: The primary purpose of this case is to demonstrate that a diagnosis of Alveolar Soft-Part Sarcoma (ASPS) is possible by fine-needle aspiration cytology (FNAC). ASPS is a rare, distinctive sarcoma of uncertain differentiation. The majority of cases occur in adolescents and young adults, in the lower extremities (most commonly in the thigh and buttock) and there is a female predominance. Although it generally follows an indolent course, many of the patients have metastatic disease at the time of diagnosis, especially in the lung, bone, central nervous system and liver. Cytogenetically, ASPS is characterized by a specific unbalanced translocation: der (17)t(X;17)(p11;q25). Materials and Methods: Herein we report a case of a 29 year-old female patient, presenting multiple bilateral pulmonary nodules in a CT scan. Additional imagiological studies showed a 7.3 · 4.2 cm mass in the left buttock. In our Institution she was submitted to FNAC of the soft tissue mass and of a pulmonary nodule. Smears stained with Diff-Quik and Papanicolaou were available for study. Tissue from a core needle biopsy of a lung nodule was also available. Results: The FNAC of the soft tissue mass was inconclusive for diagnosis. The cytological examination of the lung lesion showed haemorrhagic smears with stripped nuclei and polyhedral malignant cells, either arranged in clusters or dispersed, with prominent nucleoli and granular cytoplasm, containing PAS-positive, diastase resistant material. A diagnosis of ASPS was rendered, and it was subsequently confirmed by histopathologic examination and cytogenetic studies performed in formalin-fixed, paraffin embedded tissue (FISH). Conclusion: Owing to its distinct and characteristic cytological features, ASPS can be reliably diagnosed by fine needle aspiration in an adequate clinical-radiological context, eventually preventing more invasive diagnostic procedures. Objective: To make a retrospective study, during a period of time, of the cases, that had a cytological examination in our institute, in order to establish the types and frequency of bronho-pulmonary diseases, diagnosed by cytology, their age and gender distribution. (9.8%) -fungal infections. The malignant cases represented 30.1%-(83 cases), most of them being adenocarcinomas. In differential diagnosis, difficulties were encountered in border-line cases. Conclusion: In the cases of those who are thought to have neoplasia or show radiological changes, the cytological examination represents an important step. The cytological method of investigation has its limits that must not be ignored. The cytopathological examination is a valuable method in diagnosis of both malignant and benign tumors that are raising a lot of issues.

with the result of thyroid FNAs obtained during years 2003-2004 without cytopathological assistance. The following diagnostic categories were compared: non-diagnostic, benign, suspicious for a follicular neoplasm, suspicious for papillary carcinoma, and malignant. Results: 185 nodules underwent US FNA with on-site evaluation as compared to 203 nodules before. The unsatisfactory aspirates due to scanty material were immediately re-aspirated. The number of non-diagnostic cases was significantly lower with on-site evaluation than without (2.7% versus 12.8%, P < 0.001). In 10 cases (5.3%), additional passes were advised for cell block and immunohistochemistry. Final cytological diagnosis did not differ from rapid evaluation. Percentage of diagnostic categories was not different in the two study periods. Conclusion: Our multidisciplinary approach with on-site cytopathological evaluation reduced the number of non-diagnostic cases and helped to adequately choose the fixative for additional study. The preliminary diagnosis reduces the patient's anxiety, permits immediate evaluation in cases necessitating surgery and reduces the delay to surgery. Objective: To evaluate the relationship between chronic renal failure and corneoconjunctival findings. Materials and Methods: Conjunctival samples from 77 patients with chronic renal failure and on regular hemodialysis were taken for cytologic examination. The filter paper that was used for sample collection was the VSWP 0.25 lm pore size from Millipore. The samples were fixed with cytospray and stained with the staining procedure described by Nelson and modified by us, in order to enlarge the spectrum of diagnostic possibilities. Results: The cytologic features and numerous applications of impression cytology were evaluated. Significant calcium deposits were frequent and extensive, as well as squamous metaplasia and decreased number of goblet cells. Inflammatory cells that infiltrated the conjunctival cells were noted in 54.5% of the patients with chronic renal failure. In addition PAS-positive mucous strands were observed. Conclusion: Impression cytology offers a simple, quick and reliable method of diagnosing various ocular lesions. It has also a role in assessing conjunctival changes in contact lenses wearers or patients with dry eye and chronic renal failure. Fine needle aspiration (FNA) of the breast is a well accepted diagnostic method in detecting malignant breast lesions with great accuracy. Objective: To evaluate the contribution of FNA in the male breast lesions in organized breast screening program for the 8-year period (2000) (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) and to correlate with the histologic findings of the surgical specimens. Materials and Methods: A total of fourty two (42) fine needle aspirates of male breast lesions performed over a period of 8-years, from January 2000 through December 2008, were evaluated. Results: Fine needle cytologic diagnosis of male breast lesions was consistent with non-neoplastic lesions in 54.76% (23/42) of the cases studied (including 12 cases of gynecomastia, 3 inflammatory cases), suspicious lesions for malignancy in 9.52% (4/42) of the cases studied and in 9.52% (4/42) with malignant lesions. Biospy of the suspicious for malignancy lesions was consistent with gynecomastia in 75% (3/4) of the cases and with malignancy in 25% (1/4) of the lesions. The 100% of cytologic malignant cases were histologically confirmed and showed comparable immunocytochemical findings regarding the oestrogen and progesterone receptor status of tumors cells. Conclusion: The overall rate of malignancy in the population studied was 12%, a percentage roughly comparable with the annual occurrence of male breast cancer. Objective: To evaluate and correlate the intraoperative cytologic results of different pathologic entities with frozen sections findings and histologic diagnosis. Materials and Methods: In the present study one-hundred seventy three (173) imprints smears taken from surgical specimens submitted to our laboratory for frozen sections were examined. The cytologic smears were stained with the rapid Eosin-Hematoxylin and Hemacolor stains. Results: The intraoperative cytologic findings were completely agreed with the histologic results in 96.5% (167/173) of the cases studied. In the remaining cases we found: 2 (1.15%) false negative cytologic diagnoses, 3 (1.73%) false positive cytologic diagnoses (all suspicious cytologic diagnoses were included) and 1 (0.5%) inconclusive cytologic diagnosis (one follicular neoplasm of the thyroid gland was included). Conclusion: In the present study, the diagnostic value of cytology for rapid intraoperative tissue diagnosis is cost-effective and unequivocal with diagnostic accuracy 96.5%. Castleman's disease is a rare clinicopathologic entity and represents a morphologically distinct form of lymph node hyperpasia. Ascitic involvement by Castleman's disease is a rare complication. Objective: To present a case of Castleman¢s disease diagnosed in serous effusions. Materials and Methods: A man 47 years old with a know history of Castleman¢s disease was admitted to the hospital because of ascitic fluid and abdominal pain. The ascitic fluid aspirated at the emergency unit as well as the consequent aspirations taken, were sent to the cytologic laboratory for the routine examination. The ascitic fluid sediment smears were stained with Papanicolaou and May-Grü nwald-Giemsa. Results: The cytologic examination of smears revealed the presence of lymphocytes in different stages of maturation as well as the presence of numerous plasma cells. Immunocytochemistry for a panel of monoclonal antibodies LCA, L26, CD138, bcl2, j, k chains was positive, while no immunoreactivity for CD3, CD5 was detected. The cytologic diagnosis based on morphologic findings, immunopositivity results, and the clinical history was consistent with diffuse Castleman¢s disease in ascitic fluid. Conclusion: The cytologic diagnosis of Castleman's disease in serous effusions is a possible but rare entity and helps to the patients' support and relief from the attended symptoms of the disease. Mucoepidermoid carcinoma is a common malignant neoplasm of salivary gland in adults and children and more frequently encountered in the parotid gland. It may infiltrate the surrounding tissue, overlying skin and metastasize to the regional lymph nodes and distant organs. Objective: To present a case of mucoepidermoid carcinoma of salivary glands. Materials and Methods: A 54 year old man was admitted to the hospital with a solid, well demarcated, painless mass in the parotid gland and a retroauricular mass .FNA was performed as a primary method for evaluating these space-occupying lesions. Results: The cytologic examination of smears stained with Papanicolaou and May-Grü nwald-Giemsa showed the presence of cellular debris and mucinous background with interspersed well-differentiated squamous cells, occurring singly or in clusters, and a few clusters of columnar epithelial cells. The presence of mucus was indicated with PAS stain. Immunocytochemistry perfomed with the panel of monoclonal antibodies for cytokeratins 5/6, 8/18 was positive. Conclusion: We wish to emphasize the role of FNA cytology diagnosis as a first approach of mucoepidermoid carcinoma and region lymph nodes metastases that influences the patient's management. Materials and Methods: Nine hundred seventy two CSF specimens were obtained from an equal number of patients admitted to our hospital because of neurological symptoms during an 8 years period from January 2000 through December 2008. Ninty six (78.7%) cases regarded patients with known malignancy, followedup during and after the completion of the therapy. Twenty six (21.3%) cases regarded patients with brain lesions of unknown etiology. The CSF sent to the cytologic department was centrifuged by cytospin and stained with Papanicolaou and May-Grü mwald Giemsa. Immunocytochemistry was performed according to the cytologic findings and whenever was necessary. Results: In our study, the cytologic examination of CSF contributed to the diagnosis of eight hundred fifty (89.4%) non-neoplastic lesions and one hundred twenty-two (15.4%) of neoplastic lesions. CSF malignant diagnosis regarded ninty one (74.6%) carcinomas and thirty one (25.4%) NHL.

In the present study, the diagnostic value of CSF cytology is cost-effective and unequivocal either as a primary approach of malignancy or regarding the follow-up of the patients with malignancy. Objective: To determine the diagnostic yield of urinary cytology in the daily practice. Materials and Methods: Cytologic and histologic reports of patients, whose urine specimens were received during 2003-2004 years, were reviewed ensuring at least a two years follow-up. We included variations in histologic diagnostic terminology into low or high grade urothelial carcinoma (LG, HG, and UC) and the cytologic categories into positive/suspicious one. Cytohistologic correlations were patient-based. Results: 7320 patients were included in the study. At least one urinary cytology was reported as positive/suspicious in 186 patients: 169 (86%) were confirmed histologically (109 HG, 51 LG) while 26 (14%) were negative. Urinary cytology was negative in 7134 patients and these results were confirmed in 7003 of them by histologic biopsy or by cytologic follow-up, while 131 (2%) had at least one positive histologic evaluation (30 HG, 101 LG). The overall sensitivity, specificity, PPV, NPV were 55%, 99%, 86% and 98% respectively. Out of 291 UC found on biopsy, 131(45%) were missed by cytology, including 101(77%) LGUC. Conclusions: The diagnostic yield of urinary cytology is high for HG UC. 78% of patients with HG carcinomas were detected by cytology. Conversely, a negative result was predictive of no cancer in approximately 98% of cases. Sensitivity for LGUC was low (33%). Causes of discrepancies will be discussed.The implementation of quality assurance reviews is necessary in order to achieve a better accuracy, in spite of the theoretic limits of the method. Objective: The aim of this study was to evaluate the prevalence and the significance of urine atypical cytologic category. Materials and Methods: The histologic and cytologic follow-up of all urine cytology cases seen during a 2 years period (2003) (2004) with a diagnosis of atypical urothelial cells were reviewed. The follow-up lasted until December 2008. Correlations were patientbased rather than specimen-based. Results: We examined voided urine specimens from 7320 patients. The diagnosis of atypical urothelial cells was rendered in 472 cases (6.4%). Histologic or cytologic follow-up was available in 193 cases: 49 (25%) and 144 (75%) respectively. Out of the 49 cases with histologic follow-up 3 (6%) had negative biopsy and 46 (94%) had high or low grade urothelial carcinoma at biopsy. Out of the 144 cases with only cytologic follow-up, 124 (86%) had repeated negative cytologic controls, while 20 (14%) had persistent atypical cytology and were excluded from this work because final diagnosis was non available. In 173 examined cases, 46 (27%) had positive histological outcome and 127 (73%) were negative. Conclusions: The analysis of this series of atypical urothelial cells in urine cytology confirms the high (27%) positive outcome of this diagnostic category. In effect a significant proportion of malignant cases would be missed if atypical category would be eliminated. However an attempt should be made to keep the atypical cells rate as low as possible, in order to make the atypical category a more meaningful group, with a real risk of urothelial carcinoma. Objective: To assess the significance of qualitative signs of follicular cells atypia in diagnosing thyroid diseases and to develop an expert system working by the principle 'question-answer'.

Results and Conclusions: An expert system (software) contained the X-matrix and six standard S-matrices. The X-matrix was designed for filling with the alternative answers yes/no regarding the presence of cells with characteristic signs of atypia in the visual fields of a preparation. S-matrices characterized the main forms of thyroid diseases, such as papillary and follicular cancer, follicular adenoma, autoimmune thyroiditis, nodular colloid and diffuse toxic goiter, by their set of yes/no answers. The answers entered to the X-matrix were compared with answers of the S-matrix. Each element of each S-matrix, besides the answers yes/no, contained its weighting coefficient corresponding to a qualitative feature of a certain nosologic form. Weighting coefficient of each feature for every nosologic form was calculated by the frequency of occurrence of an atypia sign in 30 visual fields. Comparison of X-matrices with S-matrices and verification of diagnosis according to the value of a diagnostic index with a percentage probability was performed in the automatic mode. Diagnostic index was determined by the sum of weighting coefficients of features with the coincident yes/no answers, while comparing the elements of X-matrix with the corresponding S-matrix elements. The expert system displayed on the monitor the list of nosologic forms in the order of decreasing the diagnostic index values. The disease given as the first one in this list presented the final diagnosis. Background: The vast majority of invasive breast tumors are ductal and lobular carcinomas, which are similar in many respects and their histological features occasionally overlap. Distinguishing between the infiltrating ductal (IDC) and lobular (ILC) carcinomas is clinically important because of the different pattern of systemic metastases and prognostic evaluation. Few papers, however, describe immunocytochemical markers useful for differentiation of these carcinomas. Objective: The aim of our study was to investigate the expression of E-cadherin in FNA material from breast cancer patients and its relation to tumor type and other known prognostic parameters, as estrogen and progesterone receptor status and cytological grade. Methods: We examined 26 FNA smears from breast cancer patients (21 ductal carcinomas, 5 lobular carcinomas) treated in our hospital. The processing of the cytologic material was accomplished according to the ThinPrep method. Immunocytochemical detection of E-cadherin, ER and PR expression followed using specific monoclonal antibodies. Results: Our results showed complete loss of E-cadherin expression in all ILCs, while the IDCs consistently showed variable Ecadherin positivity. No association was found between E-cadherin alteration and ER, PR status of breast carcinomas. However, correlation was found between E-cadherin expression and the cytological grade of IDCs, as the intensity of staining and the percentage of cells were higher in well-differentiated carcinomas. Conclusion: We conclude from this study that E-cadherin is a useful marker to differentiate between IDC and ILC of the breast. However, further investigation is needed, to evaluate the correlation between E-cadherin and tumor grade in order to estimate its prognostic potential.

Methods: Egr-1 regulation level was examined in human glioblastoma cells lines like U373, U251, GaMG and U87-MG under extreme hxpoxic oxygenation conditions (0.1 O 2 ), reoxygenation after hypoxia for 24 and 48 hour and oxygenated conditions (21% O 2 and 5% CO 2 ) in vitro. Protein and mRNA level were detected via western blots and RT-PCR. Cells incubated for 24 hour with 100 lM DFO served positive control for hypoxia and b-tubulin and b-actin served as loading control, respectively. Results: Egr-1 was not up-regulated via hypoxic development in different glioblastoma cells in vitro under extreme hypoxic conditions (0.1% O 2 ) or reoxygenation after hypoxia, both on protein and mRNA level. Further, there was no association between Egr-1 expression and the expression of the hypoxia induced, HIF-1 a regulated genes in a population of human glioblastoma tumor specimes examined, in vitro. Conclusions: Egr-1 regulation as an answer to hypoxic development in glioblastoma, both on protein and mRNA level is not a general phenomenon. On the contrary to the previously published data, there was no hypoxic conditions influenced Egr-1 regulation. Therefore at least in glioblastoma, HIF-1 a still can be considered as a major regulator of NDRG1 under hypoxic conditions. Further extensive analysis of tumor cells from different origins under similar physiological conditions can give more comprehensive insight about the degree Egr-1 can play a role in regulating these genes expression under hypoxic conditions. In our opinion, Egr-1 regulation under hypoxia is a cell specific post translational event.

A. Syrou*, K. Kyriakou*, J. Georgoulakis † and N. Koufopoulos* *Department Of Cytopathology, St Savvas Anticancer Oncological Hospital, Athens, Greece, Department Of Cytopathology, University General Hospital Of Athens, 'Attikon', Athens, Greece Aim: The aim of the study is to determine the immunophenotype of breast malignancies by evaluating standard applied immunocytochemistry and immunohistochemistry antibodies, and to compare their results. Materials and Methods: Samples have been obtained preoperatively by Fine Needle Aspiration Cytology (FNAC), intraoperatively or/and postoperatively from 26 patients (aged between 35 and 85 years) with breast mass which was clinically or radiologically suspicious. The FNAC samples have been partially smeared onto conventional cytological slides, and stained with 'Hemacolor Giemsa'. ThinPrep Immunocytochemistry was performed to the remaining cytological material of all the samples. The expression of CerbB2, estrogen receptors (ER) and progesterone receptors (PR) by the tumor cells was studied. Results: The cytological and histological diagnoses were in accordance, and revealed 1 invasive ductal carcinoma with atypical myeloid features, 19 invasive ductal carcinomas, two invasive lobular carcinomas, one invasive mucinous carcinoma, one invasive metaplastic carcinoma and two invasive mixed type (ductal/lobular) carcinomas. The CerbB2 expression was evaluated as zero in 19 immunocytochemical (IC) samples and in 20 immunohistochemical (IH) cases; as one in five and one; as two in zero and two and as three in two and three IC and IH cases respectively. The ER expression was evaluated as 3+ in 5 IC and eight IH cases, and the PR was 3+ in six IC and seven IH cases. The few differences between the IC and IH findings were mainly rendered to low cellularity of specific cytological samples. Conclusion: ThinPrep immuocytochemistry is a fast simple and useful method to study the immunophenotypic profile of breast carcinomas. Its accuracy allows for its use in patients' follow-up and treatment planning. Objectives: (i) To review recent worldwide reports of thyroid biopsies dividing results into standard categories: benign, malignant, uncertain (atypical, suspicious, follicular) or non-diagnostic (inadequate, unsatisfactory). (ii) To assess practices at the respective institutions that may determine the incidence within the four categories.

(ii) To suggest ways to improve results. (iv) To compare the rate of cancer in the various series. (v) To emphasize the role of needle biopsy in identifying treatable cancers of the thyroid gland. Methods: Twenty-five recently published series of biopsies were evaluated with regard to method of guidance (palpation versus ultrasound), personnel, on-site assessment, and preparation of diagnostic samples. Efficiency was defined by the fraction of non-diagnostic or uncertain samples, and those series with the highest percentage of definitive diagnoses were rated highest. Results: Printed copies of the data from 25 reports reveal rates of benign diagnoses from 9% to 90%, two-thirds of malignant diagnoses below 6.5%, and indeterminate diagnoses (uncertain and nondiagnostic) between 5.5% and 75%. Conclusions: (i) Good series have in excess of 80% benign diagnoses. (ii) The best series have fewer than 10% indefinite diagnoses. (iii) Most series have a low incidence of malignant aspirates. (iv) On site assessment is associated with better results. (v) Committed individuals or teams have better results. (vi) With palpable nodules, US guidance is not necessary. (vii) The most important statistic, i.e. the % of cancer found at thyroidectomy, is not reported, but should be the true measure of accuracy of preoperative biopsy. (viii) The validity of the 'uncertain' category is questionable. Are repeat biopsies and operations justified?

evaluation of the gastrointestinal lesions. In a retrospective study we compared our results with that of forceps biopsy, since both these techniques are mainly used for the preoperative diagnosis of neoplasms within the GI tract. Methods: The last 14 months 178 patients underwent endoscopy with the clinical suspicion of malignancy. Brushing cytology was performed before biopsy and slides were prepared both with the conventional and liquid based cytology. Results: Three patients had adenocarcinoma and one squamous cell carcinoma of the esophagus, twelve gastric cancers, two lymphoma of the stomach and colon respectively, one GIST and finally in forty nine patients carcinoma of the colon was diagnosed. Brushing cytology correctly diagnosed all the malignant cases except of one gastric and two colon carcinomas, where the cytological diagnosis was suspicious for malignancy because of the rarity of the diagnostic cells.Forceps biopsy failed to diagnose one esophageal carcinoma, two gastric carcinomas and two carcinomas of the colon. Conclusion: Brushing cytology is a sensitive, accurate and eases technique for rapid diagnosis of GI lesions. The diagnostic accuracy of endoscopic BC is higher than that of FB. There was no false positive result in neither of the two techniques.Brushing cytology has a potential advantage over Forceps biopsy since it can be used to sample a relatively larger area of a lesion and a strictured, stenotic lesion, where tissue biopsy is difficult to obtain. Unfortunately, it has a limitation. If the lesion is submucosal, subserosal or in the wall and there is no ulceration, BC is unable to obtain diagnostic cells. Aim: The purpose of the study was to evaluate the cytological criteria in fine-needle aspiration biopsy and to identify their association with the basal phenotype of breast carcinoma. Materials and Methods: Fine-needle aspiration biopsy specimens and tissue sections (mastectomy specimen) from 74 cases of highgrade invasive ductal breast carcinomas were consecutively retrieved from the files of three Institutions. Breast carcinomas were studied using the tissue microarray technique, being classified into phenotypes: luminal A, luminal B, HER2 overexpression and basal. The cytological criteria for all cases were reviewed blindly by two pathologists according to five cytological criteria: cellularity, cell pattern, presence of necrosis, nucleoli, and nuclear atypia. Exact fisher test was used to test the association between cytological criteria and the phenotypes of breast carcinoma. Results: Necrosis was present in 65% of basal breast carcinomas and 31% of non-basal breast carcinomas, and that result was statistically significant, showing an odds ratio (OR) of 3.80. The basal phenotype, compared with the luminal A, showed more necrosis (OR = 6.97), present/prominent nucleoli (OR = 8.18), and cellularity more frequently (OR = 18.03). Conclusion: Necrosis, as well as present/prominent nucleoli and abundant cellularity are criteria more frequently associated to the basal phenotype of breast carcinoma. Objective: Carcinosarcoma or true malignant mixed tumor of salivary glands composed of both malignant epithelial and mesenchymal element, is extremely rare comprising 0.04% to 0.16% of all salivary gland tumors and 0.4% of all malignant salivary gland neoplasms. We present a case of carcinosarcoma in association with pleomorphic adenoma of the parotid gland first examined by FNAC, describing its morphologic features and immunocytochemical profile. Patient and Methods: A 52-year-old woman presented with a painful palpable lesion in the right parotid region that had been rapidly enlarging for 8 weeks. The imaging studies (sonography/ MRI) revealed a parotid mass and a US guided FNA was performed. The material was transferred immediately in a vial of CytoLyt Ò solution and then two slides were prepared using the Thin-Prep2000 Ò processor.The slides were Papanicolaou-stained. After the morphologic evaluation, four slides were prepared from the remaining sample in the PreservCyt Ò solution for immunocytochemical studies, using CK, GFAP, S-100 and p63 protein.

Results and Conclusions: The aspirate was cell-rich mainly consisting of cell clusters or dissociated malignant cells of epithelial origin and less mesenchymal cells displaying marked nuclear atypia. In additon, occasional benign epithelial cells, stromal material and necrotic debris in the background were noticed. On immunocytochemistry a positive expression in cytokeratin, p63 and S-100 proteins was noticed. A cytologic diagnosis of malignant mixed tumor probably in association with pleomorphic adenoma was made, assisting in preoperative planning and confirmed by the surgically resected specimen.

A. Orologa-Kalevrossoglou, A. Skenderi, K. Zachou, E. Tzartza, I. Venizelos and K. Boni Cytopathology and Pathology Department of Hippokration, Hospital Thessaloniki, Greece

Background: Uterine leiomyosarcomas account for the 1, 6% of all uterine tumors. Brain metastases are extremely rare, with only 14 cases published in the literature. Objectives: We present a case of a woman with a history of uterine leiomyosarcoma metastatic to the brain. Case report: A 56-year old woman was admitted to the neurosurgical department of our hospital with vomiting, imbalance and symptoms of intracranial hypertension. Eight months ago, a leiomyosarcoma of the uterus had been diagnosed that was confronted surgically, with abdominal hysterectomy and ovarian resection. Neither lymph node nor distant metastases were observed. The patient was treated with combined chemotherapy. Methods: A head computed (CT) and a magnetic resonance imaging (MRI) showed the presence of a single lesion in the fosse cranial posterior and edema in the fourth ventricular region. Tumor biopsy and cerebrospinal fluid (CFS) were carried out. Results: The cytology of the CFS revealed the presence of a few malignant neoplastic cells, elongated, spindle-shaped with irregular and pleiomorphic nuclei and faintly stained cytoplasm, with indistinct borders. The histological examination showed a cellular tumor, which was composed of spindle-shaped with rounded ends, coarsely granular chromatin and conspicuous nucleoli, ill-defined, basophilic cytoplasm. Immunoreactivity for vimentin, desmin or smooth-muscle actin confirmed myogenic differentiation. The cytomorphologic picture and the histological findings were consistent with the known primary neoplasm. Conclusions: The above case illustrates that brain metastasis may occur as a rare complication in uterine leiomyosarcoma. Cytology and histology contribute in the diagnosis of metastatic tumors to the brain. Introduction and Objective: Alveolar rhabdomyosarcoma (ARMS) is a small round cell tumour and represents the most frequent soft tissue tumour seen in childhood (2,3).ARMS are aggressive sarcomas and require histiotype-specific therapy (1). Accurate diagnosis of ARMS and its differential diagnosis with other small round cell tumours is of great clinical significance. Immunocytochemistry still remains, the current ancillary method of choice in the evaluation of small round cell tumours. It is so, crucial, that pathologists are aware of the variability in immunocytochemistry pattern of these entities. Material and Methods and Results: We report two recently diagnosed ARMS in two teenage girls with 17 and 10-years-old, case1 and case 2 respectively. In case1 a huge retroperitoneal mass was found. Cytologic diagnose of ARMS in a bone marrow smear was done and confirmed by FISH technique. In case 2 the presentation was of a left retro-orbitary mass and cervical lymph nodes. Fine needle biopsy (FNB) of the lymph node was performed leading to the diagnosis of ARMS. Confirmation by FISH technique was also obtained. In both cases histology was performed. Positive Immunostains for desmin, myogenin, CD56 (NCAM) and synaptophysin were found in both cases (Case1 in histological material; case 2 in FNB material). Discussion and Conclusion: This aberrant expression of a neuroendocrine marker raises potential diagnostic pitfalls, namely possible confusion with neuroblastoma, PNET or desmoplastic small cell tumor. A broad panel of immuno markers is advised and the possibility to associate cytogenetics or molecular studies provides extra tools to achieve a correct diagnosis. Introduction: EUS-FNA is an effective method for providing diagnosis in lesions not otherwise accessible for biopsying. Aim: To evaluate the utility of EUS-FNA in our hospital in the diagnosis of lesions of the gastrointestinal tract, pancreas, lung and lymph nodes. Methods: During this period a total of 59 EUS-FNA cytologies were carried out using a 19G, 22G or 25 G needle, in the presence of a cytotechnologist. Smears and cell-block were performed. We used Papanicolaou and Giemsa stains for the smears and HE and immunostains for the cell block. Diagnosis was confirmed histologically or by follow-up (clinical and imaging). Results: Lesions were located at pancreas (n = 28), gastrointestinal tract (oesophagus, stomach, duodenum, rectum and anal canal)) (n = 12), lung (n = 1) and lymph nodes (n = 18).Seventy percent of the total of the cytologies were diagnostic. The non-diagnostic cases were mainly located in the duodenum and rectum (100% and 50%), followed by the stomach (40%) and lymph nodes (33%). The vast majority of the pancreatic cases (82%) yielded to a diagnosis: adenocarcinoma, neuroendocrine neoplasia, metastatic carcinoma, cyst and inflammation. In lymph nodes the main diagnoses were metastatic carcinoma, lymphoma and granulomatous inflammation. Conclusions: In our experience, EUS-FNA is a useful method for the diagnosis of a wide spectrum of lesions, mainly when they are located in pancreas. Introduction: Estrogen receptor (ER) -alpha has been extensively studied as a prognostic and predictive marker in breast carcinoma. ER-beta has a potential role to play in the progression of breast carcinoma and the development of antiestrogen resistance. The significance of progesterone (PGR) expression in breast cancers is less recognised. The aim of this study was to evaluate the expression of PGR in fine needle aspirates (FNA) from breast carcinoma and to analyse the impact of ER alpha and ER beta expression in FNA with other prognostic parameters -cytologic grading, CerbB-2, Ki-67 and DNA ploidy. Methods: ER alpha, ER beta, PGR, CerbB-2, Ki-67 were studied by immunocytochemical methods in FNA from 38 cases of breast carcinoma. Their expressions were correlated with cytologic grading and DNA ploidy. Results: ER alpha,ER beta, PGR, CerbB-2, Ki-67 were expressed in 47%, 45%, 68%, 64% and 11% of the cases respectively. Nine of 14 (64%) of the tumors studied were diploid and 36% were aneuploid. Two-third of cytologic grade 3 tumors did not express ER alpha and ER beta.No statistical significant correlation was observed between the expression of Ki-67 and DNA ploidy with ER alpha and ER beta. However 86% of ER beta positive tumors were positive for CerbB-2 in contrast to only 65% of ER alpha positive tumors (P < 0.05). Conclusions: Demonstration of PGR is feasible by immunocytochemistry but its role as a prognostic marker is debatable. Our data indicates that ER beta positive tumors are associated with more aggressive tumors. Further investigations are necessary to better assess this. This study was supported by Kuwait University Research Grant MG01/04.

study is to assess the utility of fine needle aspirates (FNA) in detecting MPC and document any diagnostic cytological features. Methods: FNA from 47 histologically documented cases of MPC from three hospitals in Kuwait with a prior FNA were reviewed. Three cases were excluded because of insufficient material. Aspirates were categorized as benign cytology, suspicious/positive for papillary carcinoma (PC) and other neoplasms. Cytological findings were correlated with histological diagnosis. Results: Preoperative cytological diagnosis of PC was made in 35% of documented MPC cases. Tumor size ranged from 0.2 to 1 cm (mean 0.5 cm). Smears showed follicular cells in sheets and papillary configuration with pale cerebriform nuclei having open chromatin, nuclear grooves and intra nuclear inclusions. Associated ropy colloid, lymphocytic thyroiditis, microcalcification and fire flares was also seen. Site of aspiration was an important factor -in 12 of 15 cases of MPC the FNA was from the same side as that documented histologically. MPC was detected in right lobe (seven cases), left lobe (four cases), and both lobes (one case). MPC was multifocal in four cases. Of the 44 cases, the cytological diagnosis was benign (23 cases, 15.5%), hurthle cell neoplasm (four cases, 9%), Hashimoto's thyroiditis (two cases, 4.5%) and suspicious/positive for PC (15 cases, 35%). The sensitivity of cytological examination was 34.1%. Conclusion: Cytology is potentially useful but has limited sensitivity in the diagnosis of MPC. Finding tumor in aspirated material is purely accidental and when detected the thyroidectomy specimen had to be extensively sampled to find the tumor. E-sc@n is a European network in progress. Its vocation is to create ties between pathologists involved in the evaluation of serous effusions and have research projects and publications in common. It also aims at building an educative, peer-reviewed and thoroughly illustrated content in cytopathology (under construction at http:// www.escannetwork.over-blog.com). It will ease content sharing, also by non-expert users such as thesis students.The educative content will be divided into four parts: (1) 'Pathobiology' will investigate the role of the fluid tests when considering malignancy: nucleated cell count, total protein, LDH, and cytology, (2) 'Technical requirements' will address the conditions needed to agree that slides are adequate (cell fixation and concentration, elimination of RBC, cell block, LBC, and staining), (3) 'Morphologic criteria' will address the cellular and architectural features that allow accurate diagnosis, and (4) 'Molecular biology' will show that in most cases, the distinction between benign and malignant lesions can be achieved by integrating clinical, radiological, molecular and, sometimes, ultrastructural data. A questionnaire describing how samples are collected, transmitted, received and treated is being prepared. It will also address diagnostic features (how normal, reactive and malignant cells are recognized) and ancillary techniques. Pathologists will be solicited by E-mail and by means of the EFCS website (http://efcs.eu) and Cytopathology journals. An international quiz will follow the questionnaire evaluation. Objective: Diagnosis of pulmonary coin lesions needs morphological methods. Besides a tumor, tuberculosis (TB) has to be considered in most cases. An established PCR protocol for fixed specimens should be tested on liquid-based cytological specimens (ThinPrep Ò ). Materials and Methods: CytoLyt Ò fixed specimens of 45 patients presenting clinically the differentialdiagnosis TB/other mycobacteriosis and tumor were chosen. Methods for investigation were catheter biopsy (5), needle biopsy (2), transbronchial needle aspiration (1) and bronchoalveolar lavage (1). Evaluation was performed by cytololgy, Ziehl-Neelsen stain (ZN) and PCR for detection of bacteria of mycobacterium tuberculosis complex (Mtc). Results: In nine patients (20%) DNA of Mtc was found by PCR. Cytologically an inflammatory reaction was seen in all cases. In two patients a history of TB was known, furthermore one case each of silicosis respectively posttransplant situation. ZN was positive in seven of nine cases (78%). Conclusions: LBC can easily be used for detection of tuberculosis by PCR with a protocol routinely practised in pathology. Thus, by different methods peripheral lung areas can be investigated using this technique.

30 ml of CytoLyt solution for the TP method. Both smears, CS and TP were evaluated according to several parameters: satisfactory/ sufficient, or unsatisfactory/insufficient, the cytological diagnostic and some cytological characteristics. Results: From the 50 cases studied, six were unsatisfactory/insufficient in TP with one also in CS. All of the satisfactory/sufficient cases were classified as colloid nodules or cystic colloid nodules.There was concordance in the diagnoses between the two methods. The cytological characteristics observed in TP included less blood, inflammatory cells and colloid. The nuclear definition was good in 78% of TP and 24% of CS. Conclusions: After analysis of the results we can conclude that there is diagnostic concordance between the two methods. The TP method, in spite of being longer and expensive, can bring benefits for the diagnosis, especially in cases with abundant blood and/or inflammatory cells. Objective: Pericardial tamponade occurs when pericardial fluid accumulates to the point that hemodynamics are affected. There are two mechanisms that can lead to accumulation of fluid in pericardial space in cancer patients: obstruction of lymphatic drainage or excess fluid secretion from tumour nodules on pericardial surfaces. Metastasis to pericardium occurs by retrograde lymphangitic spread or hematogenous dissemination, like in lung and breast carcinomas. Metastatic mammary carcinoma is the most common tumour associated with pleural effusions in women. It may also cause ascites and, occasionally, pericardial effusions. This last one is usually a late finding in patients with known metastatic disease.The authors present an interesting case of pericardial metastasis from breast cancer without other known metastasis confirmed by imagiologic and PET analysis. Conclusion: In patients with breast cancer submitted to longterm chemotherapy, the most frequent cause of pericardial effusions are post -chemotherapy's pericarditis.Nevertheless we must consider a metastatic ethiology. The differencial diagnosis with reactive mesotelial changes could be a problem. In these cases ICC is essential. Objective: Patients with a history of cancer in another organ who develop a lung lesion must be studied considering several options such as: The lesion is benign caused by bacterial, viral or mycotic infection or post radio or chemotherapy. The lesion can be a metastasis. The most common metastasis to lung, in decreasing frequency, are breast, colon, kidney, bladder neoplasms and melanoma. Metastatic melanomas may mimic all types of malignant tumors and present difficulties in diagnosis, especially in patients whose primary tumor is either unknown or was treated many years before as a 'benign naevus'. If the cells have binucleation, very large nucleoli and/or intranuclear inclusions of invaginated cytoplasm (nuclear holes) we can suspect a metastatic malignant melanoma. However if the cancer cells are large and contain a fine, dusty brown pigment of melanin we can establish a definite diagnosis of melanoma. The authors aim to present a clinical case of melanoma lung metastasis with bronchial involvement. Materials and Methods: Seventy-three year-old women with known diagnosis of melanoma 5 years ago. Bronchial aspirate stained by Papanicolaou and 'cell block' stained by Haematoxilin -Eosin, Masson -Fontana and Immunocytochemistry technique (HMB45). Results: In bronchial aspirate we observed neoplastic cells with fine brown pigment of melanin confirmed by Masson -Fontana. The cells were positive for HMB45. Conclusion: Although melanoma metastasis to lung is usually observed, the diagnosis in bronchial aspirates is less frequent. In more difficult cases it is mandatory to do ancillary stains like Masson-Fontana and immunocytochemistry, especially if the clinical data is limited. Objective: To evaluate the diagnostic efficiency of fine-needle cytology/flow cytometry (FNC/FC) in the diagnosis and classification of non-Hodgkin lymphoma (NHL) in a series of 446 cases and to compare the results with those of previous experiences to evaluate whether there had been an improvement in FNC/FC diagnostic accuracy. Study Design: FNC/FC was used to analyze 446 cases of benign reactive hyperplasia (BRH), NHL and NHL relapse (rNHL) in 362 lymph nodes and 84 extra-lymph nodal lesions. When a diagnosis of NHL was reached, a classification was attempted combining FC data and cytological features. Sensitivity, specificity and positive and negative predictive values (PPV and NPV) of FNC/FC in the diagnosis and classification of NHL were calculated and compared with those available in the literature. Results: FNC/FC provided a diagnosis of NHL in 245 cases and of BRH in 188 cases. In nine cases, the diagnosis was 'suggestive of NHL' (sNHL); in four cases was inadequate. Histology and clinical follow-up confirmed 102 cases of NHL and detected one false-positive. In 18 cases of BRH diagnosed by FNC/FC, histological examination revealed 14 BRH and four NHL (false negatives). All nine cases diagnosed as sNHL were confirmed by histology. Statistical analysis showed 95% sensitivity, 100% specificity, 100% PPV and 93% NPV in the diagnosis of NHL. A specific subtype was correctly diagnosed in 125 cases. Statistical analysis did not demonstrate significant improvements between the present series and previous studies either in diagnosis or in classification of NHL. Conclusions: FNC/FC is a fundamental tool in the diagnosis and classification of NHL but the exiguity of diagnostic material and other technical and clinical limitations will probably continue to limit further improvement of the technique. Objective: There are evidences that intra-thyroidal CD4+, CD25+, T-regulatory (Treg) lymphocytes are decreased in response to apoptosis in patients with Hashimoto Thyroiditis. This decrease in Treg cells may contribute to the incomplete regulation of autoreactive T cells and determine cell damage. Thyroidal functional status (TFS) may differ among HT patients and is related to the cellular damage. The aim of this study has been to evaluate active (CD69+) and apoptotic (CD95+) intra-thyroidal Treg lymphocytes in different TFS of HT. Methods: Flow-cytometry (FC) was applied to thyroidal fineneedle cytology samples (FNC) in 18 patients with HT. TLI were analyzed using the following fluoresceinated antibodies: CD3, CD4, CD5, CD8, CD25, CD69 and CD95 (FAS). TFS was determined by serum TSH, FT3, FT4 immunoassays, in specific clinical settings, to classify the patients as euthyroid (14) and hypothyroid (4) . Pearson's correlation coefficient was used to evaluate possible correlations between active Treg (CD4+/CD25+/CD69+) and apoptotic Treg (CD4+/CD25+/CD95+) proportion. Results: T-lymphocytes (CD5+, CD3+) were detected in 14 cases. In these cases, CD4+ CD25+ CD69-Treg lymphocytes proportion was lower in hypothyroid than in the euthyroid patients and CD4+, CD25+, CD95+ were mainly expressed in hypothyroidism. Statistical analysis did not demonstrate associations among Treg cells proportions in the two TFS. Conclusions: FNC may be used to assess TLI in HT. Intrathyroidal CD4+/CD25+/CD69-Treg cells reduction and CD4+/CD25+/ CD95+(FAS)+ Treg increase are expression of Treg cells apoptosis and might be related to intense thyroidal damage and risk of hypothyroidism. Rationale: Obtaining specimens from children for the laboratory diagnosis of tuberculosis (TB) is challenging. Fine Needle Aspiration Biopsy (FNAB) is underutilized in the diagnosis of TB in children, despite evidence that 8-10% of children with pulmonary tuberculosis have concurrent tuberculous lymphadenopathy. Aims: To compare FNAB with gastric aspirates and other respiratory specimens from children with clinically suspected mycobacterial disease and peripheral lymphadenopathy at Tygerberg Children's Hospital (TCH), South Africa; to evaluate the ability of FNAB to provide a rapid and accurate diagnosis in children using cytomorphology, autofluorescence and Ziehl-Neelsen staining. Methods: (1) A retrospective review of all children investigated at TCH over a 4-year period where FNAB was routinely performed in conjunction with other mycobacterial specimens within 30 days prior to, or after, FNAB. The diagnostic yield and time to diagnosis (TTD) were compared. (2) A prospective study of lymph node aspirates from children with suspected mycobacterial lymphadenopathy sent for cytology and mycobacterial culture. Results: (1) In the retrospective study FNAB was positive in 45/74 (60.8%) cases while any respiratory specimens were positive in 29/ 74 (39.2%) cases (P < 0.001). The mean TTD for FNAB was 7.1 days compared to 22.5 days for any respiratory specimen. (2) In the prospective study, in 175 children the performance of FNAB was as follows: cytomorphology -sensitivity 78%, specificity 91%, positive predictive value (PPV) 93%, ZN staining -sensitivity 62%%, specificity 97%, PPV 97%; autofluorescence -sensitivity 67%, specificity 97%, PPV 97%; and culture -sensitivity 75%, specificity 100%, and PPV 100%. The aim of our study was to test chromosomal aberrations detected by fluorescence in situ hybridization (FISH) in the diagnosis of MM in effusion cytology and to explore the potential role of p16, p14 and p15 gene methylation as an alternative mechanism of tumour suppressor gene inactivation. Methods: Fifty two effusions of histologically confirmed MM and 28 benign effusions were retrospectively analyzed by multi-target FISH assay for aberrations of chromosomes 3, 7, 17 and 9p21. In case of a negative result the corresponding MM biopsy was analyzed. Methylation specific polymerase chain reaction (MSP) for p16, p14 and p15 was performed on FISH negative MM biopsies. Results: Seventy nine percent of effusions with biopsy proven MM had chromosomal aberrations with loss of 9p21 as the most common finding. All benign effusions were FISH negative. Sensitivity, specificity, positive and negative predictive values for detection of MM by FISH were 79%, 100%, 100%, and 72%, respectively. Six of nine FISH negative effusions with biopsy proven MM were also FISH negative in the MM biopsies. Four of five FISH negative biopsies showed promoter methylation in p16 and p14 as compared to one of 12 benign controls. Conclusions: FISH is a sensitive and highly specific method for the definitive diagnosis of MM in effusion cytology. In the subset of FISH negative MM, tumour suppressor genes on the chromosomal region 9p21 are often inactivated by promoter methylation. Breast carcinoma's screening in Trieste's area detected 60 women with cancer (two of them with bilateral disease) in 2008. Fortyeight cancers were diagnosed only by FNA (C5), 12 by VAB mammotome and two by Fine Needle Biopsy after undeterminate cytology. Axillary lymph nodes US was performed in 62.9% (39) of cases. They had all been diagnosed as malignant (C5) by rapid cytology during breast nodule aspiration. US detected ten doubtful axillary nodes (U3), six suspicious ones (U4) and was negative in 23 cases. FNAC was performed on all the suspicious nodes and it turned out to be positive (C5) in five out of six, which were all confirmed by histology (sentinel node protocol): five metastasis and one node negative for malignancy.7 of U3 axillary lymph nodes were aspirated: cytology was negative (C2) for all of them and confirmed by histology. Sentinel node protocol showed nine metastasis and four micrometastasis out of 49 cases which did not undergo axillary node cytology.Our study outlines cytology's accuracy to define axillary nodes' status, suggesting the possibility to extend US followed by aspiration cytology to all screening detected cases with U3/4 axillary nodes.

ous lesions. For each FNA, all subsequent core needle and open biopsies were recorded, as was the result of the definitive procedure. The cost of definitive diagnosis was calculated for each case including the cost of FNA, imaging guidance and subsequent surgical biopsy when necessary. The cost of an alternate approach using only surgical biopsy was calculated. The average costs of the FNA and surgical biopsy only protocols were compared. Results: One hundred and sixty-five primary bone tumors underwent FNA. One hundred and six FNAs yielded a definitive diagnosis. In 59 cases, FNA was insufficient for definitive therapy necessitating surgical biopsy. Aim: The aim of this study was to acess the diagnostic value of CT-guided FNAC in mediastinal masses. Materials and Methods: One hundred fourteen patients (47 males, 57 females, age ranging from 9 month to 85 years) with mediastinal masses underwent CT-guided FNAC. The clinical, radiologic, pathologic, and cytologic material was reviewed. Results: Diagnosis was possible in 105 cases (92.2%). In nine cases (7.8%) the material was insufficient for diagnosis. FNAC diagnosis were: n = 57(50%) primary neoplasms (non-Hodgkin's lymphoma,21; thymoma,12; Hodgkin's lymphoma,5; neural tumor,8; germ cell tumor,2; thymic carcinoma,1; and malignant not otherwise specified,7), n = 16(14%) metastatic tumors (lung carcinoma, 12; breast carcinoma and carcinossarcoma, 1 each; germ cell tumors, 2), n = 9(8%) suspicious, n = 21(18%) benign conditions (granulomatous disease, 4; cyst, 5; ectopic coloid goiter and splenosis, 1 each; and negative, 10), and, necrosis and inconclusive, 1 each. Histological correlation was available in 70 of 114 (61%) cases. One case was a misdiagnosis of lymphoma for small cell carcinoma. There were three false negative and no false positive diagnosis, with a sensitivity of 95% and a specificity of 100%. Conclusions: In general CT-guided FNA is a useful tool for accurate diagnosis of mediastinal masses, but can be challenging in a minority of cases due to sparse cellularity of the lesions and necrosis. Introduction and Aims: Cerebrospinal Fluid (CSF) are submitted for cytologic examination to evaluate for infections or inflammatory conditions, or a neoplastic process. The aim of this study is to review our CSF cytology results over a 4 year period. Materials and Methods: We reviewed 981 CSF samples from 864 patients, ranging from 1 month to 90 years. CSF samples were processed by cytospin. The cases were classified as: insatisfactory (only blood or not well preserved material), benign, suspicious and malignant. Either histologic diagnosis or clinical definitive diagnosis was used as the final or gold standard diagnosis. Results: Most of the cases were diagnosed as benign (n = 753, 77.2%). These included: inflammation (n = 170) and negative (n = 583). Specific infectious organisms were identified in five cases (criptococos; n = 4 and cocos; n = 1). Eleven cases (1.2%) were reported as suspicious. On further investigations, four of these were from a primary tumor (two germinomas, one glioma and one medulloblastoma) while the other seven cases were metastasis (three B-cell lymphoma, two breast carcinoma and one prostate adenocarcinoma). 96 samples from 58 patients were malignant. The primary tumors (n = 15) included: three medulloblastomas, seven gliomas, three germinomas, one choroid plexus papilloma and one rhabdoid tumor. The metastatic tumors (n = 81) were 36 breast carcinomas, 14 carcinoma of lung, four gastric adenocarcinoma, one melanoma, two rhabdomyosarcoma, two leukemia and 22 B-cell lymphoma. In 122 cases the material was insufficient for diagnosis. Conclusions: This study shows: (1) a high number of insufficient cases (12%), (2) no false positive diagnosis, (3) the majority of malignant cases were metastatic neoplasms, and the breast is the more common, (4) all the suspicious cases, proved to be malignant. In summary, CSF cytology is an accurate and reproducible approach in a well established setting, for the patients with neurological disorders. Immediate assessment optimises specimen adequacy. Introduction: Sentinel-node per-operative analysis decides whether to perform an axillary lymphanedectomy, for the surgical treatment of breast carcinoma.Scraping cytology of the sentinel-node is a simple and rapid method, whose sensitivity ranges from 65-95%. Nevertheless, in some cases a false negative result may occur. We present our experience in a series of 844 breast carcinomas. Materials and Methods: A total of 360 patients, from a series of 844, had sentinel node dissection and per-operative study by scraping, fast stain with HE and cytological diagnosis. Subsequently all the remaining tissue were included in paraffin and consecutive cuts were performed at three levels, stained alternatively with HE and immunohistochemistry with the AE1/AE3 cytokeratin cocktail. Results: Two hundred seventy nine patients had negative sentinel node confirmed in the definitive pathological study; 61 (16.9%) cases had a positive sentinel node; and in 20 (5.5%) cases we did a false negative diagnosis. Nevertheless, 12/20 only presented a micro metastasis identified by immunohistochemistry. No false positives were found. Conclusion: Cytological scrapping is a safe method for the preoperative analyses of axillary sentinel node. The percentage of false negatives was 5.5%, but most of them correspond to micrometastasis. 

were performed with a 22-gauge or 23-gauge needle. After competition of aspiration one conventional air dried smear was obtained for MGG staining in order to make a timely diagnosis of the lesion. The remaining material was immersed and rinsed into a vial filled with fixative (Cytolyt). ThinPrep slides were then prepared using the Thin Prep 2000 Processor. Immunocytochemistry staining was carried out for Estrogen Receptor, Progesterone Receptor, C-erb2, Ki-67 and p53. Results: The total number of FNA's performed with an histologic correlation was 310 cases. The cytologic material was unsatisfactory in nine cases, negative in five cases, atypical in five cases and suspicious for malignancy in 11cases. Among the 280 cases having a cytologic diagnosis of primary breast carcinoma, immunocytochemistry was obtained in 269 cases. In the rest of the cases immunocytochemical staining was performed for Estrogen Receptor, Progesterone Receptor, C-erb2, Ki-67 and p53 in 262 cases. Correlations between the IHC and ICC results in the histological and cytological samples were statistically significant for all markers (P < 0.01). Conclusion: The accuracy of ThinPrep technique in the detection of breast lesions is comparable to those of the histological evaluation, and could be of importance for the preoperative planning of treatment. Moreover it offers the potential advantage of retraining extra material for the purpose of immunocytochemical study to offer information concerning the expression of prognostic markers such as Ki-67, p53, steroid hormone receptors and C-erb2. Objective: Dysregulation of mRNA stability is important for tumor biology, leading to abnormal expression of several proteins in malignant states. HuR, an ubiquitously expressed protein of the embryonic lethal abnormal vision (ELAV) family, stabilizes certain mRNAs, allowing increased mRNA stability and protein expression. Since HuR immunoreactivity was found to be elevated in a variety of human tumors, HuR expression on breast invasive carcinoma cytological material was examined in this study. Methods: HuR expression was examined immunocytochemically on cytological smears obtained by Fine Needle Aspiration (FNA) from 83 malignant breast lesions, coming from patients diagnosed at Institut Curie. HuR positivity, overexpression (over the mean percentage value) and intensity of immunostaining were correlated with patients' age and TNM stage, tumor histological type, grade, mitotic activity, estrogen and progesterone receptor and c-erbB2 status. Results: HuR positivity was noted in 55 out of 83 (66%) examined cases and overexpression in 28 out of 55 (51%) positive ones. HuR positivity was statistically significantly correlated with tumor histological type (P = 0.026). A trend between HuR positivity and tumor mitotic activity (P = 0.051) and progesterone receptor status (P = 0.088) were also noted. Additionally, a trend between HuR intensity of immunostaining and tumor grade (P = 0.066) and mitotic activity (P = 0.063), were also observed. Conclusion: Our study for first time reports the expression of HuR on breast carcinoma cytological material. Additionally, correlations between HuR expression and clinicopathological variables important for patients' management were found. Further molecular and clinical studies are required in order to delineate the participation of HuR in breast carcinoma progression and prognosis.

A. Bondi Anatomia Patologica, Ospedale Maggiore, Bologna, Italy

In 2006, at the Venice Congress, during the meeting of the federated societies delegates, started the EFCS Web Site Project with the aim to rebuilt the web site of the European Federation of Cytology Societies. Objective: The EFCS Web Site is: an instrument to ensure connections between the federated Cytology Societies, where to find updated names and addresses of the delegates and where to publish the keynotes of each society; a newsletter for the president, the secretary and those persons involved with special commissions of the Federation; the place where to publish announcements and reviews for books or general articles on the main topics concerning cytology: a bulletin board service for the federated Societies; a discussion space for cytologists (forum) on professional themes; a mailing tool to quickly contact the European Cytologists.The site is not: a didactic tool; an atlas of diagnostic images; a server for digital slides. General Features: The internet locator (URL) will be WWW.EFC-S.EU. The URL www.efcs.org is not available (occupied by the European Federation for Company Sport); other old URLs of the Federation are redirect to the proposed URL.The official language of the site is English; the main sections are organization, activities and reserved area. The organization includes the list of European Cytology Societies, the EFCS officers' addresses, business documents. Three subsections are included in the activities: congresses, education (tutorial, QUATE area) and scientific. Some pages are reserved to people that is allowed to built the website and publish contributions from varies countries. Results: The EFCS web site has been presented to the delegates during the Madrid Congress.The first 2 years of publication have registered a poor reading rate: the average hits (single IP address) per page is 270 per year. This is the number of individual accesses to each page and it is progressively increasing. Objective: Primary thyroid lymphoma (PTL) is an uncommon malignancy, accounting for 1-5% of all thyroid neoplasms and less than 2% of extranodal lymphomas. Pathologically, most PTL's are non-Hodgkin's (NHL) of B-cell origin with Hodgkin's (HL) and T-cell thyroid lymphomas occurring rarely. This case report illustrates the advantages of ultrasound (US)-guided fine needle aspiration cytology (FNAC) and immunocytochemistry in the diagnosis of HL, manifesting as a primary disorder of the thyroid gland. Materials and Methods: The FNA smears were stained with May-Grü nwald-Giemsa method. Immunocytochemical studies were performed with monoclonal mouse anti-human CD20, CD3, CD30, CD15, ALK-1 and EMA antibodies with avidin-biotin-peroxidase complex. Results: Giemsa stained FNAC smears were cellular with a mixed population of lymphocytes, histiocytes, variable numbers of eosinophils and large atypical mononuclear and multinucleated cells consistent with Reed-Sternberg (RS) cells and variants. In our case, the RS cells were CD30 and CD15 positive and were CD20, EMA and ALK-1 negative, confirming the diagnosis of classical HL in the context of the cytological findings. A biopsy and immunohistochemical studies confirmed the cytological diagnosis of HL. Conclusion: When combined with modern immunophenotypic analysis, FNA has greatly increased our ability to diagnose NHL, thus surgical intervention is often unnecessary for the diagnosis of thyroid NHL. However, experience with FNA in the diagnosis of HL is limited because of its rarity in the thyroid. Although examination of biopsy material will be usually necessary to confirm a diagnosis of HL, when adequate material is obtained, FNA in combination with immunocytochemistry has been successfully demonstrated to diagnose HL, and thereby helping to guide subsequent clinical intervention. The objective of the study was to develop STB -based diagnostic strategy and to provide a basis for evaluation of tumor malignancy and course of disease by means of molecular analysis. Supplemen-tary molecular tests were based on an analysis of number of mRNA copies encoding proteins involved in proliferation and apoptosis.The pattern was developed on data of 316 patients subjected to STB in the years 1998-2007.Fixed in pure alcohol, dried, not stained 'reserve' cytological smears allow to obtain amounts of total mRNA (phenol/chloroform extraction by Chomczynski & Sacchi) sufficient to carry out molecular analyses. Molecular markers of proliferation and apoptosis (histone H3, p53, BCL2, BAX and FAS mRNA) were detected by RT-QPCR (Taq-Man) with use of ABI PRISM 7700 sequence detector, starters and hybridisation probes marked with fluorescent markers FAM and TAMRA, complementary to transcripts of proliferation and apoptosis marker genes. The specificity of amplification was confirmed using electrophoresis and sequence analysis (ABI PRISM 310 sequenator). Concentration profile of the genes involved in proliferation and apoptosis were better correlated with the course of disease than morphological examinations. Fixed tumor smears enable us to perform mRNA extraction and molecular examinations. Assessment of genes expression may be considered as the valuable additional marker in diagnosis and prognosis of diffuse astrocytomas. Marked morphologic diversity of gliomas is serious problem. This prompts us to develop new, more objective markers to differentiate malignancy grade and to prognosticate course of disease of particular patient.Our study included 316 patients' data, who undergone BST in last 10 years; 234 of them had primary glial tumor, 158 met criteria of inclusion into three groups according to WHO 2, 3 and 4 grade. Clinical profile of patient and tumor, macro-and microscopic evaluation along with cyto-, histological and immunohistochemical analysis of the BST material, and molecular analysis regarding indicators of proliferative and apoptotic activity were carried out by RT-QPCR technique on mRNA obtained from cytological smears. Correlation between results of the immunohistochemical and molecular analysis, malignancy of tumor and course of disease estimated in three categories: stagnation, recurrence, death, was investigated. Comparison between proteins and particular genes on one hand and course of disease on the other hand was performed in patients with follow-up period of at least 1 year. Evaluation of proliferation and apoptosis in gliomas on the basis of molecular investigations of mRNA obtained from cytological smears correlates better with course of disease than immunohistochemical examinations of biopsy material. Gene expression analysis could be valuable marker of prognosis and help to individualise the patients' therapy. Objective: To report a rare case of eosinophilic gastroenteritis associated with eosinophilic ascites. Case Report: A 38-year-old female presented with diffuse lower abdominal pain and diarrhea. She had a medical history of allergic disorders. Computed tomographic scan revealed ascites and small bowel thickening. Laboratory tests, gastroscopy, cytologic examination of the ascitic fluid and biopsies of various sites of her gastrointestinal tract, were carefully evaluated. Results: The laboratory evaluation was as follows: white blood cell count 17 500/mm 3 (eosinophils 43%), red blood cell count 4700/ mm 3 , Ht 37.6, IgE 5380 U/ml and c reactive protein 0.733 mg/dl. Upper endoscopy and cytologic examination showed normal esophagus, gastritis and duodenitis with marked eosinophilic infiltration. Colonoscopy revealed marked diffuse eosinophilic infiltration of the small intestine. Ascitic fluid cytology showed numerous mature eosinophils, macrophages and reactive mesothelial cells. Conclusion: Eosinophilic ascites is an uncommon inflammatory condition whose etiology is not well understood and is associated with eosinophilic infiltration of the gastrointestinal tract and the pancreas. It is necessary the entire digestive tract to be investigated when eosinophilic ascites is diagnosed. Background: Angiomyolipoma of the liver is a rare benign tumor composed of variable admixtures of adipose tissue, smooth muscle, and thick-walled blood vessels. Hepatic angiomyolipomas reveal morphological heterogeneity and mimic hepatocellular carcinoma or sarcoma. Their recognition is crucial to appropriate therapy. Methods: All two cases were sent for intraoperative consultation. The smears prepared from the touch imprints were fixed in 95% ethanol and stained with hematoxylin-eosin and Papanicolaou stains. Immunohistochemical stains for HMB-45 and smooth muscle actin, hepatocyte common antigen, and cytokeratin (AE1/AE3) were performed. Results: All two cases showed high cellularity. The smears were highly cellular. Spindle-shaped tumor cells were arranged in clusters, irregular bundles, or single cells. The nuclei were elongated or cigar-shaped, with finely granular chromatin and small nucleoli. There was a mild nuclear pleomorphism. The cytoplasm was granular or fibrillary. Naked nuclei were noted. Intranuclear cytoplasmic pseudoinclusions were occasionally found. Other fields revealed epithelioid tumor cells having central, round or vesicular nuclei with fine chromatin and small nucleoli. These cells were arranged in solid nests or trabecular pattern. Endothelial cells were occasionally found at the edges of the nests or trabeculae. Mature fat cells were observed in only one case. Red blood cells were seen in the back-ground. There was no necrosis. On immunohistochemical stain, tumor cells were positive for smooth muscle actin and HMB-45. Conclusions: Hepatic angiomyolipoma shows the morphologic spectrum ranging from spindle to epithelioid tumor cells. Hepatic angiomyolipoma can be diagnosed by cytologic examination if pathologists are aware of its distinctive cytologic features. Immunohistochemical stains for HMB-45 and smooth muscle actin are useful for the correct diagnosis. Objective: The evaluation of thyroid nodules using the split-sample processing on US FNA specimens and the comparison of liquidbased method with the conventional one. Patients and Methods: In all, 806 thyroid US/FNAs from 699 patients, admitted to the 2nd Department of Radiology of our hospital, were included in this study. Two air-dried May-Grü nwald-Giemsa (MGG) stained slides and one slide prepared with the ThinPrep Ò technique (Cytyc Co., Boxborough, MA, USA) were examined for diagnostic purposes. Results and Conclusions: Cytological diagnosis concerned 740 benign lesions, 34 suspicious and 32 neoplasms. In 112 cases, cytological diagnosis was made by the ThinPrep Ò slide alone, while in 14 cases by the MGG smear alone (v 2 = 20.22, P < 0.001). Five cases of neoplasms (three papillary carcinomas, one Hü rthle cell tumour and one follicular tumour) were detected by the ThinPrep Ò technique alone, while two papillary carcinomas by the MGG smear alone. Although no statistically significant difference was found between the above studied methods, as it concerns the diagnosis of neoplasms, liquid-based method alone may provide an effective tool for preparing thyroid US/ FNA specimens, offering a better quality of smears, a cytotechnical convenience in US Outpatient's Department and the advantage of further ancillary techniques on the same specimen, in which cytomorphological diagnosis was made. However, the full application of those tests from dedicated research labs to cytopathology outpatient settings has not completely been accomplished, as a number of pratical issues have not been addressed yet. The aim of this study was to assess the impact of smear adequacy on-site evaluation with respect to the rate of FNAs adequate for DNA mutation testing. Methods: This study was carried out by collecting prospectively 130 FNAs. These were routinely performed over 3 months and picked up consecutively without any selection. All aspirations were performed by a cytopathologist taking care of the immediate transfer all residual material to a vial containing RNA later (Ambion). Diff-Quik smears prepared from the first two passages by the nodule were on site evaluated; a different protocol was applied basing on the results obtained by the on site evaluation; the third pass in cases, whose cellularity had been assessed as adequate (NCI conference criteria) was fully dedicated to RNA later enrichment. Conversely the other cases (showing a not satisfactory Diff-Quick smears) had a conventional pass followed by needle rising. DNA extraction was performed from all cases by the Qiagen.DNA extraction mini kit. Results: On site evaluation led to a very low rate of inadequate FNAs (2/130; 1.5%). The group with a molecular cytopathology dedicated pass (n = 45) had an average DNA concentration of 25.9 ng/ll; the group without a dedicated pass (n = 85) had an average concentration of DNA of 7.95 ng/ll. This difference was statistically significant. Conclusions: Our data show that on site evaluation is worthwhile to increase the DNA extracted yield from thyroid aspirates. Thus in the moleculary cytopathology validation protocols on site evaluation needs to be included. Objective: In order to evaluate the advantages and drawbacks of the use of gastric wash as a diagnostic tool in patients with gastric cancer, we reviewed a series of 155 gastric washes performed, in the last 8 years, per operatively in gastrectomies of patients with previously diagnosed adenocarcinoma. Methods: From 2000-2008, 155 patients with a histological (biopsy) diagnosis of gastric adenocarcinoma were submitted to gastric washes during gastrectomy. Results: The series consists of 155 patients: 94 males (61%) and 61 females (39%), with a mean age of 65.5 years. The cytologic diagnosis was: 'malignant' in 34.8% of the samples, negative in 55.5%, suspicious for malignancy in 4.5% and non-representative in eight patients (5.1%). Correlations were performed in order to identify the relation between gastric wash sensibility and tumour morphology and histologic characteristics: tumour size, macroscopic characteristics, location, WHO/Lauré n classifications, pTNM staging and presence of atrophic gastritis. Only positive and negative results were considered (suspicious for malignancy was used as a positive result). Significant associations were found with size of the tumour (P < 0.05), WHO classification (P < 0.01) and Lauré n classification (P < 0.05). Most positive gastric washes occurred in tumours with more than 2.5 cm and when the tumour classification was other than signet ring type (WHO) or other than diffuse type (Lauré n). The presence of chronic gastritis with or without atrophy seemed to have no influence in the outcome of the cytology results. Conclusion: Gastric washing proved to be a low sensible method, and showed no advantages in the diagnosis of gastric cancer, particularly in small lesions and signet ring/diffuse type carcinomas (WHO/Lauré n). Objective: Pancreatic cancer is one of the most aggressive tumors with less than 10% long-term survivors. The majority of cases are not surgically resectable at the time of diagnosis because of early metastasis and vascular invasion. The aim of our study was to investigate the diagnostic and prognostic value of tumor markers in pancreatic tumor bearing mice. Materials and Methods: Our study included 20 female C57 black mice in which pancreatic PAN-O2 tumor was subcutaneously implanted and five mice used as controls. In 10 out of 20 mice the implanted tumor was resected at the 15th day (the 1st day at which tumor was palpable), where as in the rest mice the tumor was resected at the 30th day (day at which the size of tumor was maximum without necrosis). Then, histological examination was followed. Moreover, FNA was performed and cytological samples were processed by ThinPrep method. On the specimens from both experimental and control groups immuno-histo-cyto-chemistry was applied using a panel of diagnostic and prognostic tumor markers (mesothelin, pancreatic polypeptide, galectin-3, survivin, PCNA, . Results: The markers were over-expressed in both cytological and histological samples of mice with implanted tumor. Their expression was nearly concordant at the 10th and 20th day. On the contrary, the control samples were negative or slight positive in some markers such as PCNA, Ki-67 and pancreatic polypeptide. Conclusions: The use of these markers seems to be reliable to the diagnosis and prognosis of pancreatic adenocarcinoma. The concordant expression of the studied markers at the onset and the end of disease indicates their usefulness to the early diagnosis. Moreover, their application to cytological material provides satisfactory information contributing to the early detection of primary or metastatic tumor. Objective: BK virus has been associated with ureteral stenosis in renal transplant patients and hemorrhagic cystitis in bone marrow transplant patient. Our aim was to determine BK virus presence based on immunocytochemistry and PCR assays in urine and plasma samples. Materials and Methods: One hundred ninety urine samples derived from 32 new renal transplant recipients were studied. Cytological examination of urine samples was applied by using liquid based method (Thin Prep) for detecting decoy cells. The virus antibody JC/BK was used for immunocytochemical analysis. Real-time polymerase chain reaction (PCR-RT) was also applied for qualitative and quantitative detection of virus presence in urine and blood. Results: Decoy cells were identified in 24 cases by using cytological examination. PCR analysis detected BK virus in 14 urine samples (14/190 -7.3%). Three cases (n = 3) were positive in blood and urine samples, whereas in three other cases positive urine samples were correlated with negative blood samples and eight cases were negative in both type of samples. Immunocytochemical reaction of these samples was negative, while four positive samples were negative in both blood and urine substrates. PCR analysis also demonstrated 13.6% of the plasma samples and 25% of the urine samples as positive for BK virus. Conclusions: Concerning the detection of polyoma BK virus, PCR analysis showed higher levels of sensitivity compared to cytological examination. However, identification of decoy cells in urine cytological examination derived from transplanted patients acts as an indicator for polyoma BK virus presence. This report describes an interesting case of lung extramedullary hematopoiesis diagnosed by fine needle aspiration (FNA) cytology in a 78-years-old man presented with a left lung mass mimicking carcinoma. Thin Prep prepared, Papanicolaou stained smears of the aspirate showed numerous small and intermediate in size cells isolated without nuclear atypia and among them giant cells displaying multi-lobed nuclei resembling those of megakaryocytes. Immunocytochemistry with CD61, CD33 and hemoglobin confirmed the presence of megakaryocytes, myeloid precursors and erythroid precursors respectively. The presence of hematopoietic cells in correlation with the position of the lesion led to diagnosis of extra-medullary hematopoiesis, which was confirmed histologically. Extramedullary hematopoiesis is a rare condition defined as the appearance of hematopoietic elements outside of the bone marrow, mostly detected in chronic myeloproliferative disorders, congenital hemolytic anemias and myelofibrosis. Familiarity with these findings in the lung could be helpful in uncovering occult hematological disorders accompanied by extramedullary hematopoiesis. Extramedullary hematopoiesis should be also considered as a cause of pulmonary hemorrhage. Objective: Granular cell tumors (GCT) are currently thought to be of neural origin. They are widely distributed in the body. Salivary gland GCTs are, however, infrequent. The objective of this study is to discuss the diagnosis and differential diagnosis of fine needle aspiration (FNA) of GCT of the salivary gland. Materials and Methods: Fine needle aspiration of a right parotid mass in a 57-year-old male was reviewed. Slides were prepared by the ThinPrep methodology and stained with Papanicolaou stain. Cytologic findings were correlated with histologic features of the subsequent resected specimen. Results: The FNA was adequately cellular. There were groups of epithelioid polyclonal cells with abundant granular cytoplasm. Nuclei were small, round and regular, with a low N : C ratio and prominent nucleoli. No mitoses or necrosis was seen. Scattered inflammatory cells were seen in the background. The subsequent resected specimen along with immunohistochemical studies confirmed the cytologic impression of granular cell tumor. Important differential diagnosis includes oncocytoma, oncocytic carcinoma, Warthin's tumor, acinic cell carcinoma, and malignant granular cell tumor. Conclusion: Fine needle aspiration can be used to accurately diagnose granular cell tumors of the salivary gland. Careful cytomorphologic evaluation along with immunohistochemical studies and consideration of differential diagnostic entities are necessary to render the correct diagnosis on the FNA.

Immunocytochemistry revealed that these cells were positive for Vimentin, LMWK, CD56, synaptophycin and CD99 (MIC-2) and negative for LCA, Desmin and Chromogranin. These results led to the diagnosis of an extraskeletal sarcoma of Ewing/PNET group tumors, which was confirmed histologically. Pulmonary PNET tumors are very unusual. In these cases, the tumors appeared to arise within the lung parenchyma without evidence of bone involvement. Most patients are adolescents or young adults with symptoms of cough, hemoptysis, fever, shortness of breath and chest pain. The differential diagnosis includes other round cell sarcomas as well as epithelial and lymphoid tumors that may present as intrapulmonary neoplasms. Careful microscopic evaluation and use of immunocytochemical markers, in correlation with clinical history lead to a correct interpretation. Introduction: Fine needle aspiration biopsy (FNAB) is indicated in the study of salivary gland tumours.Due to overlap of cytological characteristics among different diagnostic entities, cystic or necrotic lesions raise differential diagnosis issues. Case Report: A 60-year-old man presented with a painless mass in his left parotid gland region that had been presented for the previous 6 months. Physical examination revealed a movable and firm mass.The patient was referred for FNAB during witch we obtained two samples, each with a small amount of creamy white fluid.In cytologic examination we observed necrotic debris along with epithelial squamous cells, some of them atypical. We could not exclude the possibility of a malignant tumour; therefore the patient underwent surgical resection of the left parotid gland. During macroscopic examination, inside of the gland we observed a well circumscribed tumour, with a partially cystic cut surface; witch contained creamy semisolid debris and small soft excrescences. The histological final diagnosis was metaplastic (or infarcted) variant of Warthin tumour, not associated with previous FNAB. Discussion: Aspiration of necrotic material in a lateral cervical nodule must raise several diagnosis possibilities:(1) Well differentiated squamous cell carcinoma (primary/secondary) (2) Branchial cleft cyst (3) Salivary gland tumour, with necrosis and squamous metaplasia The correlation with the imaging information is very important in order to define precisely the localization of the lesion. In our case, according with the imagiologic features, the lesion was inside the parotid gland, witch lead us to two possible diagnosis:

(1) Necrotic Warthin tumour with squamous metaplasia (2) Malignant tumour with epidermoid component. Even though these two entities occur rarely in the salivary glands, it is not possible to perform an accurate differential diagnosis with FNAB because of the overlap of their cytological features, witch means that frequently it is necessary to perform superficial parotidectomy of the affected gland in order to obtain an accurate diagnosis. HOW DO YOUNG AND SENIOR  CYTOPATHOLOGISTS INTERACT WITH TELE-CYTOPATHOLOGY APPLICATIONS? M. R. Giovagnoli*, S. Balzano*, S. Belmonti* and D. Giansanti † *Universita`La Sapienza, Roma, Italy, Istituto Superiore di Sanità, Roma, Italy

Objective: The development of new information technologies, the diffusion of new visualization strategies and the availability of free visualization tools have radically changed the scenery within the cytopathology laboratory. Tele-cytopathology, based on the 'virtual microscopy' and 'virtual slide' is going to play an important role in these changes. The study of the new technologies available for telecytopathology is becoming thus a basic and core issue in Telemedicine. Materials and Methods: The study focused both on student and senior cyto-pathologists to investigate how different degrees of experience in this field can influence the approach to the new technologies. Ten students and three senior experts examined six cervico-vaginal 'virtual glasses'. Diagnosis, screening time-length and acceptability were recorded. Each student was asked to select from 5 to 15 significant fields that could be used for tele-diagnosis. Results and conclusion: Our research showed that the use of tele-cytopathology applications is effective and feasible for telediagnosis. In particular, the study on how young and expert investigators approach to the new technologies showed that, in spite of te fact that they interact with 'virtual-cytology' in a different manner, all of them can reach the target 'correct diagnosis' with high accuracy. This investigation also showed other indirect and tangible advantages of 'virtual cytology' especially as a learning tool. Lymphadenopathy is one of the earliest and commonest manifestations of HIV patients. Fine needle aspiration cytology is also an adequate common procedure in the evaluation of lymph nodes in HIV patients. The most frequent aetiology of this clinical manifestation, in Western studies, is the presence of reactive hyperplasia due to the HIV itself and infectious diseases with opportunistic agents, namely Mycobacterium infection. The diagnosis of other microorganisms, such as fungi, helminthes or protozoan is less likely and most cases are reported as curiosities, escaping to most series published. We report a case of chronic visceral leishmaniasis in an HIV-1 patient where fine needle biopsy was performed in an axillary lymph node during the course of follow-up. The lymph node aspirates showed numerous macrophages, carrying several intracellular microorganisms (Leishmania amastigotes). Our patient had already been treated 2 years before because of an infection by Leishmania. The cytological diagnosis offered no major challenge, but the differential diagnosis with other intracellular infectious agents that can also affect HIV patients should always be discarded, as Histoplasma capsulatum. In this context we made a revision, of the HIV patients with lymphadenopathy seen in our hospital and submitted to fine needle biopsy in the last 5 years. From a series of 201 patients and 250 fine needle biopsy samples, this was the only case of Leishmaniasis to date. The other diagnoses performed were: reactive lymphoid hyperplasia (n = 134 -53.6%), Mycobacterium infection (n = 37 -14.8%); lymphoma (n = 10 -4%), metastatic carcinoma (n = 4 -1.6%), fungi NOS (n = 1 -0.4%) and in 61 (24.4%) samples material was considered insufficient for diagnosis. Objectives: Chordoma is an uncommon malignant neoplasm arising from persisting notochordal remnants. It is usually located in the axial skeleton. We report a case with its cytologic and histologic correlation. The aim of the study was to assess the usefulness of FNAC (fine-needle aspiration cytology) in the diagnosis of metastatic chordoma. Materials and Methods: Forty-six year-old female presenting with a history of recurrent chordoma in thoracolumbar region and an abdominal mass. FNAC was performed on the abdominal mass. Diff-quick and Papanicolau staining protocols are also used as well as immunohistochemistry. Results: Microcopic examination of the smear showed moderate to scant cellularity arranged in lobules or cords. Cells were large in size and vacuolated (physallipherous). There were also smaller tumor cells with a polygonal to a fusiform shape. All the cells were immersed within a myxoid-appearing extracellular matrix. The tumor cells were immunoreactive for EMA and S100. Objective: O6-Methylguanine-DNA-methyltransferase (MGMT) is an enzyme that interferes in the repair of damages caused by endogenous and environmental alkylating agents, by removing the alkyl groups. Abnormal MGMT expression has been reported to correlate with prognosis in patients with solid tumors. MGMT expression on cytological material and its validation in clinical cytology have not previously been referred. Methods: MGMT expression was examined immunocytochemically on cytological smears obtained by Fine Needle Aspiration (FNA) from 61 malignant breast lesions, coming from patients diagnosed at Institut Curie. MGMT positivity, overexpression (over the mean percentage) and intensity of immunostaining were correlated with patients' age and TNM stage, tumor histological type, grade, mitotic activity, estrogen and progesterone receptor and c-erbB2 status. Results: MGMT positivity was noted in 31 out of 61 (51%) cases examined and overexpression in 14 out of 31 (45%) positive ones. MGMT overexpression and intensity of staining were statistically significantly correlated with tumor size (pT) (P = 0.022 and P = 0.027, respectively). MGMT intensity of staining was also statistically significantly correlated with c-erbB2 status (P = 0.017). Additionally, a trend between MGMT positivity and tumor histological grade (P = 0.092), and between MGMT intensity of staining and estrogen receptor status (P = 0.068) was also noted. Conclusion: Our study for the first time reports MGMT expression on breast invasive carcinoma cytological material. Additionally, correlations between MGMT expression and clinicopathological variables important for patients' management were found. Further molecular and clinical studies are required in order to delineate the participation of MGMT in breast carcinoma progression, prognosis and treatment. Objective: The differential diagnosis of thyroid lesions is critical for clinical management and sometimes is difficult to determinate diagnosis, in Fine Needle Aspiration (FNA) specimen. The immunocytochemical expressions of Cytokeratin 19 (CK-19) and CD-56 were evaluated to asses their potential as markers in the diagnostis of Thyroid lesions. Materials and Methods: Fine needle aspirates of 70 patients aged from 13 to 81 years, (mean 56.9), were performed under ultra sound guidance. The material aspirated was used on conventional smears as well Thin-Prep preparation and was analyzed by immunocytochemistry using monoclonal antibodies Cytokeratin-19 and CD-56.The neoplastic cases included 28 carcinomas (24 Papillary CAs, two Follicular CAs, two Medullary CAs) and six Adenomas (four Hurthle cell, two Follicular). The non neoplastic Thyroid lesions included 36 cases (26 Nodular Hyperplasia and 10 Hashimoto's thyroiditis). Results: CK-19 expression was found in 21 of 24 Papillary CAs, one of two Medullary CAs, in 10 of 26 Nodular Hyperplasia and in 3 of 10 Hashimoto's thyroiditis.Cytokeratin-19 expression was absent in two of two Follicular CAs and in all Adenomas. CD-56 expression was found only in 1 of 23 Papillary CAs, in one of two Medullary CAs, in one of two Follicular CAs, in four of six Adenomas, in 5 of 10 Hashimoto's thyroiditis and in 20 of 26 Nodular Hyperplasia. Cytology has been for many decades one of the most valued tools for diagnosis. Years of practice have shown that the accuracy of cytology varies with many factors starting with the quality of the samples, the experience of the cytologist and also the difficulty in interpreting cellular changes in some tissues more than others. The authors, experienced cytologists and clinicians, feel that cytology of mammary tumors in dogs and cats is hardly useful as a diagnostic tool, essentially due to the fact that the determination of malignancy grade, especially in dogs, is highly subject to error. This renders cytology, a highly precious diagnostic procedure, low utility specifically in mammary pathology. To gather evidence of the previous statement, correlation between fine-needle aspiration cytology and histopathology was performed in 37 samples of mammary tumors in dogs and 14 samples in cats, as follows: agreement in ten cases of malignancy was achieved, both in cats and dogs; accurate diagnosis of mammary tumor with no added predictive value was achieved in 33 out of 34 cases, posteriorly diagnosed histopathologically as benign tumor (3), grade 1(12), grade 2(9) and highly malignant grade 3(2) carcinomas, and in the cat seven malignant tumors were confirmed in a total of eight cases. Cytological diagnosis of benign lesions was by far the most difficult, with agreement achieved only in two out of five cases. The results show that cytology is not a useful diagnosis tool in clear cut mammary tumors before surgery, but should be the first choice in case of doubtful neoplastic lesion, with no individualized nodules, in cases of recurrence or if there is lymph node enlargement. Needless is to say that histopathology should be performed in all circumstances as the golden standard of diagnosis, allowing the establishment of prognosis with high accuracy. , 41% of the nodules were single. The mean age was 73.4 and 52 in primary tumors (13 males and 8 females) and in patients with metastatic nodules, respectively. Results: Single nodules were founded in 62.5% of primary tumors and in 33.3% of metastatic patients. Regarding echogenicity, primary tumors were ipoechogen in 37.5% and metastatic nodules in 52% of the cases. The smears presented the similar amount of cells in 72.5% of cases. With the exception of two cases that showed to be inadequate for one method, all the diagnoses performed on LBC, were confirmed on the corresponding CB. Morphologically a better nuclear detail was evident in LBC whereas structural features were appreciated on CB. Immunocytolchemical assay was performed on 92 cases: 17 (18.5%) on the monolayered smears and 75 on the CBs according to more availability of the latter material. Conclusion: In malignant primary or metastatic hepatic nodules, FNC on LBC evidenced a similar diagnostic accuracy as compared with CB technique even though a smaller amount of cells useful for immunochemistry was available. P.398 SALIVARY GLAND CYTOLOGY R. Cameron Pathology/Cytology, Lund, Sweden

The value of aspiration biopsy cytology (ABC) in the clinical evaluation of masses of the neck is well established. The efficacy of ABC in the diagnosis of salivary gland lesions has been a point of discussion for many years with different case reviews making different conclusions. Pitfalls in the cytological diagnosis of salivary gland tumors have been well described. If these pitfalls be recognized and avoided, false ABC diagnoses should be minimized.To determine if increased experience leads to improved diagnostic quality, we evaluate ABC diagnoses made in salivary gland tumors, which were operated during 2 time periods at a regional referral center in southern Sweden. The 2 periods are 1989-1998 and 1999-2008 . Squamous epithelial cancers, melanom metastases and malignant lymphomas were eliminated from the evaluation. Approximately 40 cases of salivary gland tumors of other types were operated each year during this time period and essentially all of these were diagnosed using ABC preoperatively. We evaluate the original ABC diagnoses and correlate these with the histopathological diagnoses, determining diagnostic accuracy, sensitivity and specificity. The 2 time periods are compared. In cases where discrepancies between ABC and histopathological diagnoses are discovered, slides are reviewed to see if known pitfalls have been at play and these are discussed. The diagnosis of amyloid goiter by fine needle aspiration (FNA) secondary to this predisposing disease has not been described before. Objective: To illustrate two cases of amyloid goiter secondary to RA diagnosed by FNA in private and academic settings. Material and Methods: A 33 year-old woman with RA, renal amyloidosis, adrenal insufficiency and diffuse enlargement of the thyroid was submitted to FNA and had alcohol-fixed smears sent to a private cytopathology laboratory in Brazil. Slides were initially stained by Papanicolaou technique and re-stained by Crystal Violet solution. A 31 year-old man with longstanding juvenile RA and chronic renal failure presented at an academic hospital with diffuse symmetrical enlargement of the thyroid. Prior kidney biopsies were positive for amyloid. Slides were stained by the Papanicolaou and Congo red methods. The 2nd patient died of sepsis 3 years after diagnosis. The thyroid gland at autopsy weighed 370 grams due to marked amyloid deposition. Results: Abundant cyanophilic to orangeophilic amorphous material was observed in the Papanicolaou stained slides. Bundles of elongated stromal cells, occasionally with artifactually stretched nuclei, were found interspersed in the material. Rare groups of normal follicular cells were also identified. Amyloid was revealed respectively by Cristal Violet and Congo red stains in each case. Histological sections obtained in one of the cases showed abundant eosinophilic, congophilic amyloid substance deposited in the interstitium surrounding dilated atrophic thyroid follicles. Conclusion: FNA proved to be an effective tool for diagnosing the very rare entity of amyloid goiter secondary to RA in two distinct clinical scenarios. Attention to the clinical presentation and morphology of the cells accompanying amyloid allows exclusion of medullary thyroid carcinoma, avoiding unnecessary surgery. Objective: To compare the cytologic diagnoses of breast lesions obtained with fine needle aspiration (FNA) technique when applying liquid-based cytology to the results of histopathologic material. In addition we compared the diagnoses of fine needle aspiration obtained by Thin Prep (TP) technique with those of conventional smears in order to correlate the advantages and disadvantages of both techniques. Material and Method: The study includes 548 FNAs performed on women presenting a clinically palpable breast mass. One conventional air dried smear was obtained for MGG staining in order to make a timely diagnosis of the lesion. The rest of the material was immersed a rinsed into a vial filled with fixative (Cytolyt) and ThinPrep processed smears were then prepared using the ThinPrep 2000 Processor for cytomorphologic diagnosis. Aspirates were classified as: inadequate, benign, atypical-probably benign, suspicious probably malignant and malignant. Histologic diagnosis was based on biopsy or a mastectomy specimen. Results: In this study ThinPrep FNA of breast lesions shows a sensitivity of 98.49%. The specificity of fine needle aspiration was 100%. The positive predictive value was 100% and the negative predictive value was 98.41%. Comparing conventionally prepared smears to ThinPrep technique the percentage of unsatisfactory aspirates was decreased, from 12.04 to 6.39% respectively. Finally a good correlation was observed among TP, conventional cytology and final diagnosis (conventional cytology versus TP:0.90, Histology versus conventional cytology: 0.97 and Histology versus TP: 0.98). Conclusion: The application of liquid based cytology (ThinPrep) on fine needle aspiration is an accurate technique for the cytomorphological evaluation of breast masses prior to surgery or follow-up. Objective: The purpose of this study was to evaluate HPV DNA test in relation to cytomorphology for the management of women after conization. Material and Methods: Our study included cervical samples of 56 women (24-56 years old) that underwent conization for HSIL (CIN 2 or 3). The specimens were processed by Liquid Based method (ThinPrep technique). After Pap-staining and cytological examination, on the rest material of each case HPV testing was realized by the Hybrid Capture 2 system. On 7/56 cases hc2 and PCR (Clinical arrays, Genomica) were applied before and after treatment in order to study the alteration of viral load and to detect the responsible for the recurrence HPV types. Results: The evaluation of the results was conducted in relation to the age of women, the cytomorphological findings and the results of molecular methods (HPV DNA test and PCR). The mean age of women who underwent conization was 35.9 ± 7 years. According to cytomorphological lesions (ASCUS, LSIL, HSIL), 32.1% of patients developed a recurrence after a mean follow-up of 34.9 months. Based on findings of both of cytomorhology and HPV DNA test, residual disease was observed in 21.4% of patients after a mean follow-up of 19.2 months (± 17). Moreover, at the post-treatment period the viral load was gradually decreased. Finally, the results of PCR showed the HPV types 16, 31, 33 and 51. Breast angiosarcoma (AS) is a rare aggressive tumor and few cases diagnosed by FNA have been reported. A case of breast AS with no radiation history is presented.A 58 years old woman with a prior diagnosis of breast angiomatosis a year before presented with a protruding mass of the same breast with a bluish-red discoloration of the overlying skin. FNA of the lesion yielded heavily bloody material. Giemsa and Papanicolaou smears and cell block were prepared. Microscopically spindle cells with elongated moderately atypical nuclei and epithelioid cells in tight clusters with papillary configuration were observed. Paraffin sections of the cell block included tissue fragments composed of anastomosing vascular channels with endothelial proliferation and solid areas of spindle cells. Immunocytochemistry showed positivity of spindle and epithelioid cells to endothelial markers CD31 and CD34. The differential diagnosis included recurring angiomatosis and AS. The cytological diagnosis was cellular vascular neoplasm with nuclear atypia suggesting AS. After total mastectomy the histological diagnosis was high grade AS. Retrospective histological evaluation of the 'angiomatosis' lesion was consistent with low-grade AS. Three months after mastectomy the patient presented with two dermal masses, at the forhead and the anterior chest wall. FNA of both lesions showed metastatic involvement from the breast AS. In conclusion FNA of breast AS yields very bloody material (reflecting the wide vascular channels) with low cellularity. To prevent false negative results cell block preparation is helpful often containing tissue fragments entrapped in blood clots. Predominance of epithelioid cells and papillary structures may suggest epithelial tumor, especially in patients with a history of breast carcinoma and predominance of spindle cells may raise the possibility of phyllodes tumor or other sarcomas. Immunocytochemistry is required for the correct diagnosis. Objective: Fine Needle Aspiration is a reliable, safe and costeffective procedure widely used not only in the primary diagnosis of neoplasias, but also in accessing their progression. We reviewed a five year period of FNAC performed during the follow-up of patients with a previous diagnosis of neoplasic disease that developed a new superficial palpable tumor in our hospital. Material and Methods: Between 2004 and 2008 we performed 154 FNAC in the standard manner in patients with nodes in areas of surgical scars (56) or others superficial sites (98), on chemotherapy (47) or just in follow-up (107). The material was processed as smears or liquid-based cytology and stained by Papanicolaou stain. Results: The primary tumors had different origins, types and evolution times. Ninety-four (94) FNAs were negative, with foreign body granulomas in 21 cases. Sixty FNAC, were positive for malignancy with seven being recurrences on scars, forty five metastases in lymph node, six in soft tissue and two new neoplasias. The recurrences and metastasis had the same morphological type of the primitive tumor, with 19 cases positive under chemotherapy and 39 on follow up. Twenty one were from breast carcinomas, seven carcinomas from gynecological sites, 14 from gastrointestinal tract, three urological, four from prostate, seven from haematological sites and four from skin and soft tissues. Conclusion: FNAC is a useful method to evaluate progression of neoplasic diseases and adds value on distinguishing systemic metastasis from new neoplasias, and recurrence from negative or inflammatory nodules. Objective: Laminin-5 is a heterotrimeric extracellular matrix (ECM) molecule normally present in the basement membrane as part of the epithelial adhesion complex. The detection of the laminin-5c2-chain in cytoplasm is a primary pathological event indicating the ability of tumor cells to invade.In tumor tissue the laminin-5c2-chain is considered as a guide for invasion. Our study investigates laminin-5c2-chain immunocytochemistry as a tool for the identification of carcinoma cells in pleural effusions and ascites.

Materials and Methods: Non-tumorous and malignant effusions were analysed using immunocytochemistry: laminin-5c2-chain, clone D4B5, delution1:10000, streptavidine/alkaline phosphatase methodology (DAKO, Denmark), DAKO autostainer. Results: In cells of non-small cell lung cancers a regular cytoplasmatic immunostaining was seen in cytospins. The inflammatory cells and mesothelial cells were not immunolabelled. In ascites adenocarcinoma cells showed a regular laminin-5c2-chain positivity in the cytoplasm. Negative immunostaining of laminin-5c2-chain was regularly observed in cases of small cell lung cancer and breast cancer. Non-epithelial cells like malignant melanoma cells were not identified by laminin-5c2-chain immunostaining. Conclusions: The laminin-5 immunocytochemistry is a very reliable tool for identifying non-small cell lung cancer cells and abdominal adenocarcinoma cells in pleural effusions and ascites. A positive laminin-5 immunocytochemistry was not observed in small cell lung cancer cells and breast cancer cells. In conclusion, laminin-5 contributes to cytological differential diagnostics in effusions. Objective: Vascular endothelial growth factor (VEGF) is a multifunctional cytokine.This secreted endothelial cell-specific growth factor constitute an important angiogenic pathway which under hypoxic conditions, is up regulated in many solid tumors such as breast and lung cancer. Recent bibliographic data report that VEGF is overexpressed in laryngeal SCC. The purpose of this study is to investigate the expression and diagnostic value of VEGF in cytological material of laryngeal squamous cell carcinomas. Materials and Methods: We examined 30 smears (negative: five, suspicious: seven, positive :18) obtained by brush from patients with laryngeal lesions. The material was processed both using the conventional method and Liquid Based Cytology (ThinPrep Ò ) technique, immunocytochemical analysis was followed using VEGF monoclonal antibody (clone VG1) Diagnostic Biosystems,Ylem. Results: Positive expression (cytoplasmic staining) of VEGF was observed in the majority of squamous cell carcinomas and suspicious cases. In addition some of the tumor-infiltrating inflammatory cells showed also VEGF expression in many of the carcinomas studied. All negative smears revealed no expression of VEGF. Conclusions: VEGF is overexpressed in the majority of laryngeal squamous cell carcinomas and could be used as a possible diagnostic marker of malignancy in the everyday clinical practice. We investigated the expression of hTERT mRNA using ISH method in 66 fluid cytology samples from 60 patients including 36 pleural fluids, two pericardial fluid, 24 peritoneal fluids, and four bronchial washing. Comparisons of the results of ISH with conventional cytological evaluation were also performed. Cytomorphological examination of 66 body fluids was classified as malignant, suspicious, atypical and benign conditions. Of 34 malignant conditions, there were 30 cases both positive by cytology and ISH. There were 18 discrepant cytology-ISH results in patients with varying conditions including four cases positive by cytology and inconclusive by ISH, 10 cases suspicious by cytology and positive by ISH, and four cases atypical by cytology and negative by ISH. Of 18 benign conditions, there were 16 cases negative by both cytology and ISH. But there was two case negative by cytology and positive by ISH. From the results of the expression of hTERT mRNA using ISH method, this test was highly sensitive (sensitivity, 90%) and specific (specificity, 91%), and had favorable positive (95%) and negative (83%) predictive values. Expression of hTERT mRNA component can be easily detected in malignant cells by ISH method in fluid cytology. This method is especially useful when the abnormal cell population in fluids consists of limited suspicious cells that cannot be completely differentiated from reactive mesothelial cells and thus may help identify the true-positive cases from the pseudo-negative effusions. This ISH method for detection of expression of hTERT mRNA component may contribute an ancillary test for early recognition of cancer cells in body fluids and thus has potential as a diagnostic adjunct in cytopathology. Result: From 1017 FNAC, 808 were adequate, 209 were insufficient, 347 positive for MC, and 461 miscellaneous entities.From 347 MC, patients ranged from 29 to 88 years, 213 male and 134 female. In 89/347with known primary tumor, 76/89 the aspirates confirmed the primary malignancy, in 13/89 another primary tumor was identified. The most frequent radiological and clinical presentation: lytic lesion, pain and nodules/masses. The most frequent aspirated site: spine, pelvis bone, ribs and femur. Of MC cases, carcinoma with different subtypes 178(51%), 152(43.8%) were adenocarcinomas, 10(2.73%) were small cell carcinoma (neuroendocrine) and 8(2.3%) undifferentiated tumors. Primary tumor site: breast, lung, pancreatic and gall bladder, kidney and prostate in order of frequency. Conclusion: Using thicker-gauged needles has improved the technique obtaining satisfactory samples, avoiding insufficient specimens and reaching high diagnostic accuracy. If previous malignancy is unknown, performing IHC on CB is determinant establishing the origin of primary tumor. CK7, CK20, CK19, PSA, Hormonal Receptors and TTF-1 are the most useful markers. On FNAC specimens, cytomorphology combined with IHC viewed in a clinical-radiological context, obtains a diagnostic accuracy rate over 95%. Objective: The primary purpose of this study is to determine whether a diagnosis of Hyalinizing Trabecular Adenoma (HTA) can be made solely on the basis of the cytologic features observed in this case. Materials and Methods: Two cytospin Papanicolaou-stained slides prepared from specimen labeled 'fine needle aspiration (FNA) of right thyroid' containing 2 ml. of cloudy, blood-tinged fluid. Two cytospin Papanicolaou-stained slides prepared from specimen labeled 'fine needle aspiration (FNA) of left thyroid' containing 2 ml. of clear colorless fluid. Results: Smears from FNA of the right thyroid showed clusters of follicular cells with atypical nuclear features including intranuclear cytoplasmic inclusions and nuclear grooves. Some of the follicular cells show papillary configuration. Psammoma bodies and reactive lymphocytes were also noted. Final cytologic diagnosis was Positive for Malignant Cells, features are consistent with Papillary Carcinoma arising in a background of Chronic Lymphocytic Thyroiditis. Smears from FNA of the left thyroid showed small clusters and sheets of uniform bland follicular cells, occasional Hurthle cells, reactive lymphocytes and muscle fibers. Final cytologic diagnosis was consistent with Chronic Lymphocytic Thyroiditis. Conclusion: Follicular cells in clusters and papillary configuration with nuclear grooves, intranuclear cytoplasmic inclusions and psammoma bodies seen in FNA of the right thyroid are features found in both Hyalinizing Trabecular Adenoma and Papillary Thyroid Carcinoma. Other diagnostic features of HTA such as amorphous hyaline materials and cytoplasmic yellow bodies were not observed in the smears. Therefore, the cytologic diagnosis of Hyalinizing Trabecular Adenoma cannot be made based solely on the cytologic features seen in these smears. Objective: We report four cases of skin non-Hodgkin's lymphomas suggested by fine needle aspiration cytology (FNAC) in which PCR based clonality detection was failed although a B cell clonal population was demonstrated by Flow Cytometric Immunophenotyping (FCI). All cases histologically were diagnosed as cutaneous marginal zone lymphomas. Material and methods: Aspiration smears from cutaneous nodules was done for cytomorphological diagnosis. FCI was performed on aspirated material diluted in PBS with application of a panel antibodies (CD3, CD4, CD8, CD56, CD19, j and k). PCR was performed on liquid specimen (PreserveCyt solution) for amplification of Fr2/J region of the IgH heavy chain. Results: Cytologically, in two cases predominant small cells appear on smears and in the other two, a mixed small and large cells were observed. In all cases, j or k light chain restriction were revealed in FCI. Clonal IgH rearrangements have not been detected in neither of the four cases. Conclusions: Cases of non Hodgkin's lymphomas without clonal IgH rearrangements detection in molecular investigation does not exclude B clonality. Application of FC immunophenotyping in FNA material may confirm monoclonality and is a useful adjunct to approach cutaneous marginal zone lymphomas. Objective: Metanephric tumors (MT) (Metanefric adenoma ((MA) and metanefric adenofribroma (MAF)) are rare benign tumor of kidney. Occur at any age, young female 2:1 preponderance and rarely in children. They have good prognosis, not associated to malignancy, although metastasis have been reported. Most cases are asymptomatic and present as an incidental finding but clinical features such as haematuria, fever, hypertension and occasionally hematologic disorders may be present. It has been suggested that MT represent the differentiated counterpart of Wilm's tumor, although its true origin is still unknown. We report 2 cases in pediatric patients.

Material and Methods: Case 1: 12 year-old female and Case 2: 1 year-old male. Both of them presented an incidental mass without hematologic, hormonal or clinical special features only a pielic secondary ectasia. Fine needle aspiration (FNA) was performed in both cases. Cytological material was available (Dif-Quick and Papanicolau stains) and cell-block obtained in case 1 allowed an immunohistochemical (IQ) study. Subsequent tumorectomy was done. Results: Cytology highly cellular tumor, with pseudo papillary structures composed by rounded small cells, with scanty pale light cytoplasm and rounded-oval uniform nuclei with homogenous chromatin and absent or inconspicuous nucleoli. In IQ staining on cell-block, tumour cells were positive for WT1 and CAM5.2 stain. Ki67 was expressed in 10 to 15% of tumour cells. Histology revealed a tumor with occasional branching, elongated tubules, and papillary structures. Blastema was absent and stroma ranged from inconspicuous to a loose edematous appearance. Numerous psammoma bodies were present. Histological diagnosis of MA and MAF was established. Conclusions: MT are extremely rare tumor in children. Cytological diagnosis with FNA is possible due to specific morphological features but is important to use a wide panel of IQ stains in order to make a correct diagnosis. In children and young patients differential diagnosis has to be established with renal cell carcinoma papillary type, Wilm's and neuroectodermal tumour, in order of their different behaviour and worse prognosis. Objective: Fine needle aspiration (FNA) of the thyroid gland has been accepted as a first line screening test for patients with thyroid nodes. Currently, a cythologic diagnosis into six categories has been proposed.We report our experience in fna with correlation with cytologic-histologic findings in 100 cases. Methods: All cases were examined by means ultrasound-guided FNA. A cytopathologist was always present and both Diff-Quick and Papanicolau staining methods were used. The findings were classified as follows: unsatisfactory, bening, atypical cellular lesion, follicular proliferation lesion, suspicious of malignancy and positive for malignancy. Results: 1% of the lesions were classified as unsatisfactory; 63% as bening; 9% as atypical cellular lesions; 12% as follicular proliferation; 1% as suspicious for malignancy and finally 14% as to be malingnant. The cytologic-histologic correlation were 100% in malignant cases and 99% in bening cases. Six out of nine cases classified into the category of atypical cellular lesions were nodular hyperplasia; Two out of nine were follicular adenoma and the remaining was a malignant lesion. Among the atypical cellular lesions, one third required surgical removal. Five out of 12 cases of follicular proliferation were nodular hyperplasia, four out of 12 were follicular adenoma and finally, three out of 12 were malignan lesions. Hence, the number of lesions that actually required surgical removal were seven. Conclusions: The current results indicated that FNA provides an accurate diagnosis of thyroid pathology. The six categories were beneficial to define patients for either clinical follow-up or surgical management. Aim: The aim of the study was the comparative correlation between androgen receptor (AR) expression and tumoral grading in urothelial carcinoma on cellular and tissular blocks. Material and Methods: The studied material consisted of 15 cases diagnosed with urothelial gall bladder carcinoma. Preoperative vesical washing with isotonic solution was performed and the sediment was embedded in parraphin wax in order to obtain cellular blocks. Preoperative tumoral biopsies were also performed and tissular fragments were also embedded in parraphin wax. Hematoxilin-Eosin stain was used for basic histological investigation on sections obtained both from cellular and tissular blocks. AR within urothelial cells was immunomarked with specific antibodies (clone AR441, DAKO, 1:50 dilution) both on cellular or tissular blocks. The following pathological features were assessed: type of urothelial carcinoma (grading, according to WHO classification), intensity of AR immunohistochemical expression (using the following quantitative score: 1 -low intensity; 2 -moderate intensity; 3 -high intensity) and density of the AR expression (using the following quantitative score: 1 -<25% positive cells; 2 -26-50% positive cells; 3 ->50% positive cells). Results: The majority of studied cases were poorly differentiated, 66% of them being in G2 stage. The comparative analysis between AR expression on cellular and tissular blocks revealed that, on well differentiated tumours (G1), AR intensity was strongly expressed on both cellular and tissular blocks, with a higher value on tissular blocks. Similarly, AR density was higher on tissular blocks. On poorly differentiated tumours, AR intensity was almost similar on cellular and tissular blocks, although some cases revealed higher expression on cellular blocks. The score was, however, lower than that in well differentiated group, no 3 score being reported. Similarly with differentiated tumours, AR density was higher on tissular blocks in a significant number of cases. In the only one case of grade 3 (invasive) carcinoma both the intensity and density of AR positive cells had the same expression on cellular and tissular blocks, with a score of 1. The Pearson index had a significant value (P < 0.05) for both correlations between AR intensity on cellular and tissular blocks and AR density on cellular and tissular blocks. The index had a high significant value (P < 0.001) for correlation between AR intensity and density on both cellular and tissular blocks. Conclusions: These preliminary study revealed that AR is present on tumoral cells but with no correlation between its intensity and density expression and tumor grade. On the other hand, the one necrosis.Histological confirmation revealed three false negative and no false positive cases. The false negative cases were: one insufficient, one necrosis and one desmoid fibromatosis, that proved to be, respectively desmoid fibromatosis, metastatic adenocarcinoma and dermatofibrosarcoma protuberans. Conclusion: This study shows: 1-The majority of subcutaneous masses of the abdominal wall subject to FNAC are neoplastic, and 2-FNAC is an effective technique for primary assessment of these lesions. Introduction: Urine cytology has long been a useful tool for the diagnosis of urinary tract cancer as well as for detection of metastases in the urinary tract. The presence of positive urine cytology in patients with lung cancer without obvious urine tract metastases is not expected. In literature only a few cases have been reported (especially non Hodgkin's Lymphomas) of patients harboring malignant cells in urine cytology without radiological verification of metastasis to the urinary tract. Patients and Methods: It is an ongoing study and up to now 140 cases of patients with lung cancer (Small and non Small Cell Lung Cancer) have been studied. The urine cytology of 15 patients with metastatic colorectal cancer, 10 with non Hodgkin's lymphoma and 15 with metastatic breast cancer have been studied up to now as control group. The diagnostic methods used in our study are the following: (1) Three voided urine samples at 3 consecutive days evaluated by liquid-based cytology,(2) Immunohistochemical Staining, (3) CAT Scan (CT) of the urinary tract and (4) Cystoscopy with bladder biopsy. Results: Twenty of the 140 patients (14%) had positive urine sediment cytology. None of these patients had radiological verification of metastasis to urinary tract. The morphological appearance of the cells coming from the biopsy and the urine cytology were identical. Three patients underwent cystoscopy with bladder biopsies which were negative for metastases. The urine cytology of the patients in the control group were negative. Discussion/Conclusions: Urine cytology in collaboration with immunohistochemical staining and molecular techniques can help in the detection of disseminated cells in the urine, a phenomenon that has not been studied in detail until today. Large, retrospective or prospective cohort analyses are needed in order to study its true incidence, correlation with clinicopathologic tumour characteristics and patient management as well as its predictive and prognostic significance. Purpose: Alkaline-encrusted pyelitis are rare unrecognised lesions on urinary cytology. In order to emphasize the skillness of the cytologists, the authors report a characteristic case with review of the literature.

A 67 year-old woman admitted to our hospital with a completely destroyed right unique kidney was treated with immunosuppressor for an auto-immune hepatitis. She has previously presented hematury. On TDM, neoplasic lesion was suspected. Results: Three urinary cytologies coming form washing procedure were very difficult to interpretate. On a cytolytic background, cells are mostly reduced to sand aspect with some dense hyperchromatic residual nuclei and lot of debris but urothelial cells are devoid of atypia. The examination with polarized lumen showed peculiar squared phosphate ammoniaco-magnesien crystals called struvite stones. The opportunity to examine the ureteronephrectomy performed for persistant macroscopic hematury and elevated creatinemia provided the solution of the diagnosis by both histology of an encrusted pyelitis and bacteriology showing the infection of the urinary tract by Corynebacterium urealyticum. Urine tests show an alkaline urinary pH, struvite stones, leucocytury and hematury. In fact the ammomia odor is strongly indicative. The infection is under diagnosed resulting to the difficulty to isolate C. urealyticum from urine so the cytology is important. The literature is mainly composed of case reports or short series. The lesion is also described in dogs and cats. Increasing numbers of cases are diagnosed, especially in immunodepressed patients, and particularly in renal transplant recipients. The clinical informations are of major interest for the diagnosis. The clues are struvite stones. Objectives: Metastatic lung adenocarcinoma is a common cause of malignant pleural effusion and its early diagnosis is important for planning the management of patients with lung tumors. The expression of TTF1 by the neoplastic cells confirms the diagnosis of pulmonary adenocarcinoma and helps to exclude other mimics. Immunohistochemical testing for TTF1 expression is usually performed on paraffin embedded cell blocks, which may not be possi-ble to prepare from limited samples. In this study we tested the utility of TTF1 immunocytochemical staining in liquid based cytology preparations. Materials and Methods: Thinprep cytology slides from 80 consecutive malignant pleural effusion with adequate samples were immunohistochemically stained for TTF1 along with sections of the corresponding cell blocks. All slides were stained by a Ventana automated system using streptavidin-biotin kit after heat-induced epitope retrieval. The stained Thinprep slides were independently assessed for the pecentage of stained nuclei and the intensity of the staining by three of the authors. Each parameter was assigned a score of 0, 1, 2 or 3 and the total score of both parameters (a minimum of 0 and a maximum of 6) was calculated for each case. A total score of three or higher was considered a positive result. The results were compared to that of the immunostained sections of the corresponding cell blocks and also to the final diagnoses. Result: The neoplastic cells of 19 (23.8%) of 80 collected samples were TTF1 positive on the liquid based preparation. There were seven false negative and one false positive specimens. For metastatic pulmonary adenocarcinoma, the specificity, sensitivity, positive predictive value and negative predictive value of TTF1 immunostaining on the monolayer cytology preparations were 98.1%, 73.1%, 95.0% and 88.3% respectively. Conclusion: In liquid based cytologic preparations, TTF1 is a sensitive and a highly specific marker for metastatic pulmonary adenocarcinoma. Thinprep slides are a reliable alternative for immunohistochemical staining, when paraffin embedded cell blocks are not available. Objective: To investigate the feasibility of the newly device preparation of cell block using polybrene as a mediator for research and cytological diagnosis and compare with the conventional method. Material and Methods: We used the polybrene (hexadimethrine bromide) as a mediator and binder which is a quaternary ammonium polymer carrying four positrons for cell clotting and congregation. Body fluids and three different cancer cell lines (KB, HeLa, OEC-M1) were used as testing samples for establishing the newly device preparation of cell block and to compare this modified cell block with conventional method for application of research and cytological diagnosis. Results: It takes less cell numbers for preparation of polybrenemediated cell block than the conventional method. Our newly device preparation of cell block with polybrene may provide a densely packed, relatively smaller cell block with a clean background and even cellular distribution. It may also offer a satisfactory platform in both immunohistochemistry for differential diagnosis of cytology and in situ hybridization in vitro study. Conclusion: The newly device polybrene-mediated cell block is an easy and cheap way for yielding a high quality and compact cell block, as well as offers a useful and reliable method for research and cytological diagnosis much better than conventional method. PCNSL is extranodular lymphoma, developing within CNS, most frequently in deep brain structures. It do not causes symptoms of systemic disease. Contemporary standards of histological diagnostics of PCNSL involve stereotactic biopsy followed by cytologic, histologic and immunohistochemical investigations of bioptate.We analysed 202 patients' data subjected to stereotactic biopsy (Brain-Lab system) of deep CNS tumors in last 8 years. Volume of bioptates ranged from 0.5 to 1mm3. Routinely stained smears (Harris hematoxylin) were prepared for intraoperative cytologic diagnostics. Special attention was paid to possibility of establishing diagnosis on basis of cytological smears. Remaining material underwent histological and immunohistochemical investigations (CD45, CD79a, CD20, CD3, CD2, CD30, pan CK). In 22 cases establishing diagnosis on basis of of cytologic smears was difficult or impossible, because of significant cell necrosis. Histological and immunohistochemical investigations enabled us to diagnose 32 PCNSL: 26 B-cell, 3 T-cell, 1 B-cell-T-cell rich and 2 anaplastic large-cell lymphoma according to WHO classification. Cytologic diagnostics of stereotactic bioptates of PCNSL is difficult, because steroids given before surgery to prevent herniation, cause necrosis of texture and impede evaluation. Histological and immunohistochemical of very scarce material obtained by stereotactic biopsy in patients with PCNSL enables us to establish diagnosis and implement appropriate therapy.

ANGIOSARCOMA OF THE THYROID GLAND -CYTOLOGIC DIAGNOSIS AND DIFFERENTIAL DIAGNOSIS L. Pisharodi Rhode Island Hospital and Brown University, Providence, RI, USA Objective: Angiosarcoma of the thyroid gland is a rare entity. The objective of this study is to describe the cytomorphologic and immunocytologic features of this neoplasm and to discuss the differential diagnostic possibilities. Materials and Methods: Fine needle aspirations (FNA) of thyroid and the lung were reviewed. Slides were prepared using the Thin-Prep methodolgy and stained using the Papanicolaou stain. A cell block was also prepared. The patient had presented with shortness of breath. CT scans revealed a large mass in the thyroid and multiple lung masses. In addition to the FNAs, subsequent core biopsies and autopsy material were reviewed. Results: FNAs of the thyroid and pleura showed similar morphologic features. Individual cells were oval to round, cytologically atypical with a high N:C ratio, hypochromatic nuclei and prominent cherry red macronucleoli. Majority of cells were 'epithelioid', Objective: A 3-gene (PCSK2, PLAB, CCND2) qPCR based assay was reported to have high accuracy for distinguishing benign from malignant thyroid tumors that are often indeterminate on fine-needle aspiration (FNA) biopsy (1). More recently PCSK2 and CCND2 were confirmed to be useful, whereas PLAB gave discordant results (2). Our group has previously shown that UbcH10 and HMGA1 are novel diagnostic thyroid cancer markers (3) (4) . In the present study UbcH10 and HMGA1 were added to PCSK2 and CCND2; the diagnostic performance of this 4-gene assay was evaluated. Material and Methods: qPCR was applied to tissue surgical samples from normal (n = 10) and neoplastic (n = 106) thyroid in a training set. Subsequently, qPCR was applied to NON NEOPLASTIC ALSO neoplastic FNAs (n = 20) with tissue follow-up. The target genes were PCSK2, CCND2, UbcH10 and HMGA1. The nonparametric Mann-Whitney Rank test was used to determine differences between mRNA expression levels. A P value of <.05 was considered statistically significant. To determine the diagnostic accuracy of the 4-gene assay, the area under the curve (AUC) of the receiver operating characteristic (ROC) curve was determined for each gene individually and in combination by the logistic regression analysis. Results: Our results confirmed that PCSK2 and CCND2 are differentially expressed between normal and neoplastic tissues. We also assessed that these targets may be reliably evaluated on FNAs. The diagnostic accuracy was increased by the including into the panel UbcH10 and HMGA1. Conclusion: A 4 genes assay has diagnostic usefulness increasing the diagnostic performance of PCSK2 and CCND2 qPCR detection. Preliminary results suggests that 4 gene assay is also feasible on FNCs UbcH10 and HMGA1 are useful diagnostic thyroid cancer markers, as the 4-gene assay has a good diagnostic accuracy. P.429 PRACTICING, TEACHING AND LEARNING CYTOLOGY -PERSONAL EXPERIENCE AND ADVICE E. Sze´kely, R. Istó k, T. Sze´kely and B. Ja´ray 2nd Department of Pathology, Semmelweis University Budapest, Hungary

Objective: Many doctors deny the reliability of cytological examinations. The reason for this phenomenon is, that both smear taking, preparation, and interpretation of the smears needs skill and experience. Unfortunately, the number of enthusiastic cytologists is low. Many doctors give up the learning period early, in the lack of satisfaction and feed back.

Material and Methods: Learning cytology in a tutorial model helps to improve all the abovementioned steps quickly. In our department we have about 2500 ultrasound guided fine needle aspiration cases (liver 330, pancreas 50, kidney 40, retroperitoneum 20, breast 420, thyroid 550, lymph nodes 450, salivary glands 100, other sites 500) yearly, 10% of them are sent for immuncytochemical reactions as well. On certain occasions, fluorescent in situ hybridization is also performed on specifically prepared smears. Two-thirds of the samples are taken by the cytologist, with much better effectivity: (3.5 /20% inadequate smears in case of lymph node aspirates, taken by cytologist or clinicians, respectively). H&E staining of the wet-fixed slides gives opportunity to compare the smears with the histological picture of the same lesion, after surgical resection. Our residents participate both at the consulting hours, and join the seniors during reporting. Later on they can practice the sample-taking with the guidance of the seniors as well.In the recent period we started to improve our teaching 'repertoir' by suggesting the residents taking scrape smears from the fresh surgical resection specimens, on arrival in the laboratory, and during autopsy. The interpretation of the smears becomes more exciting for them, since the smears are comparable with the histology of the lesion in two days. Conclusions: Teaching cytology might replace this diagnostic method to it's deserved position, for the patients' benefit. In all of them FNAB was done. 374 patients were female and 101 male, aged 20-88 years, and most of them were 50-69 years old.Slides were stained by May-Grunwald-Giemsa, air dried and analysed under light microscope. Results: In 217 patients cytological diagnosis was hyperplasia, adenoma in 71, and in 187 there was not sufficient material for cytological analysis. Only in 36 of those 187 inadequate samples, parathyroid hormone (PTH) was positive in the aspirate. On echosonography, adenomas were presented in most of the cases as hypoechoic, solitary nodule and in 84% located behind lower pole of the right or left thyroid lobe.In 96% of cases we were able to diagnose adenoma of the parathyroid gland, based on examination of MGG stained slides, with additional staining for argyrophil reaction. Based on cell morphology we were able to distinguish oncocytic adenoma of parathyroid gland, atypical adenoma and chief- Objective: Cytometric quantitation of nuclear DNA content could be significant for assistance in the diagnosis and grading of malignant tumors. However while most laboratories are equipped for digital imaging not all of them have the necessary software and personnel for DNA ploidy analysis. The aim of this study was to compare the results obtained by the ploidy analysis of digitized nuclei images performed by on-site versus remote cytopathologists. Material and Methods: Nuclei images from smears prepared using ThinPrep Ò method and stained according to Feulgen technique have been captured using a digital camera. A general image analysis software (Image Pro Plus) was customized for image processing, nuclei segmentation and DNA-ploidy analysis. Appropriate nuclei were selected by an on-site cytopathologist. Histogram of IOD has been drawn and parameters describing Ploidy status such as Ploidy Balance, Proliferation Index, DNA Index, 2CDI, Malignancy Index and DNA Malignancy Grade have been calculated for each case. After compressing the captured images using the jpg standard at 60% compression level, they were sent through internet to a second cytopathologist who independently performed DNA-ploidy analysis using the same customized software. Results: A good correlation was observed comparing the results obtained from each user of the image analysis software (r > 0.85). Conclusions: Provided that the imaging equipment meets the least standard specifications and the appropriate procedures are followed for image capturing and processing, DNA ploidy analysis could be performed in a remote specialized laboratory for assistance in the diagnosis as well as for inter laboratory agreement sustainability reasons.

MALIGNANT SEROSAL TUMOUR, UNUSUAL PRESENTATION W. Salman and Z. Twaij East Lancashire Hospitals, Lancashire, UK Objective: We present a case of primary malignant peritoneal mesothelioma with the unusual immunohistochemical profile, desmin positive, EMA negative, and wish to highlight the importance of cautiously interpreting immunohistochemistry profiles when they do not fit the clinical history and histological appearances. Material and Methods: A 64 year old man with a past history of asbestos exposure and a clinical diagnosis of ascites with omental caking underwent ascitic fluid sampling. Three smears and clots were received which showed groups and single atypical mesothelial cells. These cells in the fibrin clot were positive for calretinin, CK 7, CK20 and Desmin . They were negative for EMA, Ber EP4, CK 5/6 and TTF-1. A report was issued stating that the cytology was suspicious of malignancy and biopsy of the omental cake was requested. Several fragments of omental tissue were received. Histology showed infiltration by tumour cells forming solid sheets of epithelioid cells and glandular structures with cellular atypia, nuclear enlargement and nucleoli. The tumour cells were strongly positive for desmin, CK7, calretinin, NSE and WT-1 but negative for EMA, HBME-1 and CK5/6. This was reported as being consistent with abdominal mesothelioma. Two months later the patient died and a post mortem examination was performed. The peritoneal cavity contained 1.5L of straw coloured effusion and almost all of the mesentery and omentum were replaced by tumour. The liver was cirrhotic and there were multiple bilateral pleural plaques. Tissue from the tumour and lungs was taken for histological examination and asbestos fibre analysis. Sections from the tumour showed an identical histology and immunohistochemical profile to that of the previous biopsy. Lung tissue for asbestos fibre count contained 1.47 million asbestos fibres per gram implicating asbestos exposure as the likely cause of the tumour. The cause of death was reported as malignant peritoneal mesothelioma. Paraffin blocks from the omental tumour were sent for electron microscopy. They showed abundant intermediate filaments (tonofilaments) supporting the diagnosis of mesothelioma. There have been several studies comparing the immunohistochemical profiles of benign and malignant mesothelial cells. Unfortunately there is no immunohistochemical stain that, per se, allows separation of benign from malignant mesothelial proliferations. Therefore the diagnosis of malignant mesothelioma may be supported by the use of a panel of immunohistochemical stains. These include markers that are commonly expected to react with these tumours (positive markers) and markers that are not commonly expected to react with these tumours (negative markers). Most studies agree that EMA and desmin appear to be the most useful markers in distinguishing benign from malignant mesothelial proliferative lesions. EMA is preferentially expressed in up to 80% of malignant neoplasms and desmin in up to 85% of benign mesothelial lesions. However a recent review found EMA and desmin to both have a low sensitivity & specificity of 70-75%. Results and Conclusion: To our knowledge this case of peritoneal mesothelioma is the first to describe an opposite immunohistochemical profile to that expected, being EMA negative and desmin positive, despite macroscopic and microscopic evidence of malignancy. We suggest that EMA and desmin, whilst being helpful when distinguishing malignant mesothelioma and benign mesothelial cell proliferation, should not be considered infallible when the clinical, cytological and histological appearances suggest an alternative diagnosis. Objective: Transbronchial Needle Aspiration (TBNA) is a minimally invasive procedure that allows sampling of hilar and medias-tinal lymph nodes during bronchoscopy, which is important for the diagnosis of sarcoidosis stage I, since the later involves mesothoracic lymph nodes. In our study we analyze four cases and we present a review of the relevant literature. Materials and Methods: Four patients with mediastinal lymphadenopathy without parenchymal pulmonary lesions underwent bronchoscopy. TBNA was performed with sampling from the right subcarinal lymph nodes station. Also, bronchoalveolar lavage (BAL) sample was taken. Cytologic material was directly smeared on slides, fixed in 95 0 alcohol and stained with Pap stain. Transbronchial Lung Biopsy (TBLB) was not perfomed in our cases. Results: Cytologic findings form our four patients were consistent with sarcoidosis, showing nonnecrotizing granulomas in all cases and rare multinucleated giant cells in one case. Culture and acid fast stain for tuberculosis bacilli were negative. A definite diagnosis of sarcoidosis stage I was established by the combination of cytologic findings with clinical and radiological picture as well as the results of flow cytometry from BAL. According to recent literature, TBNA shows high diagnostic value with sensitivity rates ranging from 53 to 72% in the diagnosis of sarcoidosis stage I. Also, in 23-42% of the cases TBNA diagnosis was the only means of rendering a morphologic diagnosis. Finally, the combination of TBNA with TBLB increases the diagnostic yield up to 83%. Conclusion: TBNA is valuable in the diagnostic approach of patients with suspected sarcoidosis stage I, since it highly improves the diagnostic yield without any complications for the patient. The practice, knowledge, information and technological advances in cytology have gone a long way since 1970's. Cytology today has become an integral part of Medicine. Cytology investigations in Goa have also increased in leaps and bounds from 1564 patients in 1984 to 6483 patients in 2007.In Goa FNAC is indispensable in many conditions, in Tuberculosis, when patients are anxious to know whether the lesion is benign or malignant, in elderly patients, in recurrent lesions, Metastatic lesions etc. In Goa FNAC is used in superficial organs as well as deep seated organs using Ultrasound or CT scan. Various interesting lesions encountered in Goa of the thyroid, Breast, Salivary glands, Lymph nodes, soft tissues, Lungs, Mediastinum, Liver, Spleen, Kidneys, GIT, Orbit will be presented. Objectives: Salivary gland tumours are uncommon and unique in their morphological complexity, variability and pattern overlapping and therefore are a challenge for cytologists. The purpose of this study is to asses the value of FNAC in differentiation inflammatory and reactive process, benign or malignant tumour and if possible to render a specific diagnosis. Material and Methods: Within 7 years 81 FNAC of minor salivary gland lesions have been performed by a cytologist alone or in a team with maxillofacial surgeon. FNAC was performed using 23G needle attached to 20 ml syringe. The smears were stained with Papenheim, Papanicolaou and Hemacolor rapid staining. The cytological diagnoses were compared with histopathological ones. Results: We acquired adequate material from 72 (88.9%) patients. A total of 49 patients underwent surgery: 28 with benign and 21 with malignant tumours according to the histopathological diagnoses. The most common tumours were pleomorphic adenoma among benign and adenoid cystic carcinoma among malignant tumours. The most commonly affected site was the palate. The team-work of cytologist and surgeon enables specificity of 89.3%, sensitivity of 95.2%, positive predictive value of 87%, negative predictive value of 96.2% and diagnostic accuracy of 91.8%. Conclusion: It is very important to know whether the lesion is malignant or benign for planning surgery and may prompt or postpone decision for surgical intervention. Therefore, FNAC of minor salivary gland lesions should be considered in the first line investigations of these lesions. Introduction: The presence of sporotrichosis in subcutaneous tissue is not a very frequent entity. The early suspicion by cytologic examination of FNA specimen of subcutaneous lesion give us a great help to guide the clinical management. Objective: A case of sporotrichosis into subcutaneous tissue primarily hypothesized by FNA specimens is presented. Material and Methods: The 54 years old male patient was admitted to the hospital because of the presence of a 5cm · 1cm palpable subcutaneous lesion on the anterior thoracic wall with a strong clinical suspicion of malignancy. The patient was occupied as a municipal nursery gardener, was a heavy smoker and heavy drinker. He lived together with his dog that was recently deceased. He was referred for thoracic CT with the result of solid lesions in the Material and methods: All consecutive patients who underwent a EBUS-FNA between January and November of 2008 for mediastinal or hilar lung lesions, and had a diagnosis of non malignancy were included. Rapid on-site evaluation of the specimens obtained was performed in all cases, by means of Diff Quik-stained air-dried smears. Papanicolaou stains and cell blocks were also done. Results: There were 32 diagnosis of benignancy out of 63 EBUS-FNA procedures. A specific diagnosis was achieved in 6 cases (2 tuberculosis, 3 sarcoidosis and 1 tyroglossal duct cyst). The other 26 had normal lymph node cellularity. In 5 cases further study was performed due to high suspicion of malignancy (mediastinoscopy in 3 cases and esophageal ultrasound-guided FNA in 2). Benignancy was confirmed in 4 of them (normal lymph node in 3 and sarcoidosis in 1) and a squamous cell carcinoma was diagnosed in one case. Conclusions: EBUS-FNA is a useful technique for the diagnosis of non-neoplastic pathology presenting as hilar or mediastinal lymphadenopathy. In our series, 19% of cases diagnosed corresponded to sarcoidosis, tuberculosis or thyroid gland pathology. Objective: Prophylactic administration of a quadrivalent HPV6/11/ 16/18 vaccine is 98-100% effective in preventing HPV16/18-related CIN2/3 or AIS. We report the impact of the vaccine on the inci-dence of all Pap tests, colposcopy and definitive therapy, regardless of causal HPV type. Materials and Methods: 17,622 women aged 16-26 were enrolled in 1 of 2 randomized, placebo-controlled trials. Vaccine or placebo was given at Day 1, Month 2 and 6. Pap testing occurred at Day 1 and every 6-12 months for up to 48 months. Colposcopy referral was algorithm-based. Definitive therapy referral was per standard of care. We estimated number of events prevented annually per 10,000 vaccinated women, in terms of risk difference (RD) based on subtracting the rate in the vaccine arm from the rate in the placebo arm in: 1) an unexposed population that approximates sexually naïve females; 2) a mixed population of HPVexposed and unexposed women; and 3) the population of women already exposed to HPV. Results: Within~3.6 years, we observed reductions for all endpoints. In the unexposed population (defined as DNA negative at baseline to all of 14 HPV types, seronegative to HPV6/11/16 and 18, and with a normal Day 1 Pap), the annual estimated number of events prevented per 10,000 vaccinated women was: any Pap abnormality = 138 (RD = 1.38, 95%CI: 0.8, 2.0); any colposcopy = 135 (RD = 1.35, 95%CI: 0.8, 1.9); and any definitive therapy = 58 (RD = 0.58, 95%CI: 0.4, 0.8). In women who were exposed to HPV (defined as DNA positive at baseline to any of 14 HPV types and/or seropositive to HPV6/11/16 or 18), the estimated number of events prevented was similar to the unexposed population: any Pap abnormality = 148 (RD = 1.48, 95%CI: 0.4, 2.6); any colposcopy = 134 (RD = 1.34, 95%CI: 0.2, 2.4); and any cervical definitive therapy = 66 (RD = 0.66, 95%CI: 0.2, 1.1). Conclusions: Our data suggest that whether we offer HPV vaccine to a population of women who are HPV naïve, a mixed population of naïve and exposed (data not shown), or a population of previously exposed women, we could expect to have similar public health impacts in terms of reduction in abnormal Pap tests and procedures in the years immediately following vaccination.

There are few studies concerning the application of Liquid Based Cytology in breast lesions. The aim of this study is the direct comparison between monolayered (LBC) and conventional (Cv) in breast Fine Needle Cytology (FNC) and with the corresponding histological control. Morphological (cell size, nuclear features, cytological grading) and prognostic and predictive factors (PPFs) of breast cancer have been considered. An analogous observation was also performed on Core-Biopsies (CBs) on patients who underwent radical surgery. Materials and Methods: A total of 43 FNCs, performed in breast cancer patients, entered this study. Two conventional and one monolayer smears (stored in Preserv Cyt and processed by Thin Prep) were obtained, stained according to Papanicolau method. Moreover, PPFs (ER, PgR, MIB-1 and HER/B2-Neu) were detected on residual material in LBC and on the corresponding surgical material.Twenty one CBs were also analyzed for PPFs determinations and compared with the results obtained on histological controls. Regarding pathological parameters on 43 evaluable cases, 36 histological cases were infiltrating ductal carcinoma; 11 were <1 cm sized and 16 cases didn't present metastates. on fine needle aspiration biopsy (FNAB) & core needle biopsy procedures at the clinic with first visit to the Hospital. Material and Methods: Total 21 cases were selected from the computerized pathology report files. FNAB cases (16) of 'lumps & bumps' in the neck & parotid glands and core needle tissue biopsies of the lesions in the jaw bones, nasopharynx, & orbit (5), were evaluated. All cytology preparation of direct smears stained with H&E, DQ, & Pap-stain. Core needle biopsies & cell blocks were stained with H&E. The poorly differentiated malignant tumors with unknown primary sites were stained with selected immunohistochemical (IHC) markers on cell blocks

The neck region: metastatic carcinomas with known primary sites

& with unknown primary sites (3), & Hodgkin's lymphoma

Others: spindle cell carcinoma of the maxillary sinus (1), poorly differentiated squamous cell carcinoma, left maxilla (1), calcifying odontogenic cyst, left mandible (1), plasmacytoma with sinonasal adenocarcinoma & sarcoidosis, nasopharynx (1) & meningothelial meningioma

Conclusion: The neck lesions of 'lumps & bumps' diagnosed based on characteristic tumor cell morphology with FNAB-technique, was sensitive, accurate, cost effective, & offers rapid diagnoses

Greece Objective: The most common chemotherapy for colorectal carcinoma remains 5-FU administration after surgery. 5-FU targets thymidylate synthase (TYMS), whose expression is correlated with the polymorphism rs34743033. Newer therapeutics includes EGFR blocking antibodies and short molecules that inhibit EGFR activation. Even though new therapeutic strategies are available only some patients will benefit from them, since EGFR blocking is abolished if activating K-Ras mutations are present. The aim of our study was to verify whether, through pre-oparative colonoscopic brushings, molecular characteristics of colorectal lesions could be examined. Materials and Methods: Material from 123 brushing was collected in Cytolyt. A fraction was routinely processed and a slide was stained according to Papanikolaou, while the rest of the sample was used for DNA extraction. Diagnosis was set by a cytopathologist. TYMS polymorphism was examined by routine PCR. Codons 12 and 13 of exon-2 of K-Ras were examined for mutations by either sequencing or with a Real-Time PCR commercial assay. Results: One hundred and thirteen cases were cytological diagnosed as carcinomas and 10 cases as adenomas

The TYMS 5 un-translated region polymorphism's distribution was: 2 Repeats (23.6%), 3R (35%), heterozygous 2R/3R (30.9%), 2R/3R with LOH (6.5%), 2R LOH (2.4%) and 3R LOH (1.6%). Presence of K-Ras mutations was independent of TYMS polymorphism status

These tests can, in theory, predict patient's reaction to chemotherapy, assisting the clinical physician in therapy choice. Furthermore, targeted neo-adjuvant chemotherapy could aid in turning untreatable tumors to treatable.This research project (PENED) is co-financed by

Thin Prep Pap Tests in correlation with cytological morphology. Materials and Methods: Cervical smears of 320 cases with ASCUS or SIL diagnosis were analyzed using the HPV Clinical Arrays genotyping (Genomica) after PCR DNA amplification. ASCUS diagnosis was based on hyperkeratosis, parakeratosis, and megacaryosis alone or in combination. SIL diagnosis was made only when koilocytosis and or dysplastic cells where present. Results: HPV was detected in 97/185 (52.43%) ASCUS, 118/133 (88.72%) LSIL and 2/2 (100%) HSIL (CIN2) cases. High risk HPV was found in 79/97 (81.44%) ASCUS and in 106/120 (88.33%) SIL cases. Multiple hrHPV types were present in 30/97 (30.92%) ASCUS and in 57/120 (47.5%) SIL cases. The same HPV types prevailed with similar percentages in both groups: in ASCUS high risk HPV51 and HPV 53 (31.95%), HPV16 and HPV 18(13.40%), and low risk HPV6 (16.49%), and in SIL high risk HPV53 (28.33%), HPV16, (21.66%), HPV51 (20.83%), HPV31 (15.83%), HPV66

A 49-year-old woman presented for hirsutism, deep voice and hypertension. Ultrasonography (US) revealed a solitary tumor mass, 8 cm in size, of the right adrenal gland. Laboratory tests showed it to be a hormonally active, androgen secreting tumor (elevated testosterone level), which was consistent with the clinical picture of the disease. Histopathologic analysis pointed to adrenocortical carcinoma (ACC), a low risk carcinoma according to Weiss' classification. On regular follow up US study performed at 1 year, a 65 · 43 mm growth was diagnosed in the lower pole of the right kidney. The finding was verified by computerized tomography and the patient was reoperated on. Exploration revealed secondary growth in the region of greater omentum, without infiltration of adjacent organs. Histopathologic analysis indicated metastatic ACC. At 8 months of the second operation and after six chemotherapy cycles according to EAP protocol, enlarged para-aortic lymph nodes and a node along the upper pole of the right kidney were diagnosed by cytologic puncture. Cytologic analysis showed numerous large tumor cells (individual and in papillary clusters) with polymorphic, hyperchromic, irregular nuclei, prominent nucleoli with perinucleolar halo, and abundant pale basophilic cytoplasm. The cells were positive for vimentin, NSE, synaptophysin and epithelial markers (BerEP4 and epithelial membrane antigen (EMA)). Conclusion: ACC is a rare tumor of high malignant potential, deriving from adrenal gland cortex. Morphologically (histopathology and cytology), there is the problem of differential diagnosis from adenoma on the one hand, and from renal cell carcinoma (RCC) and hepatocellular carcinoma (HCC) on the other hand. Proliferation markers such as Ki67 or DNA-ploidy help differentiate ACC from adenoma, and cellular markers (alpha-fetoprotein, CD10, RCC marker, vimentin, EMA, keratins 7 and 20, alpha-inhibin, and polyclonal antibodies against carcinoembryonic antigen (pCEA)) from RCC and HCC. Objective: The aim of the study was: (1) to assess the appropriateness and adequacy of lymph node puncture for flow cytometry analysis in case of benign and primary malignant lymph node lesions, and (2) to assess the value of cytologic analysis in various groups of malignant and benign lymph node lesions by determination of B lymphocyte clonality using the method of flow cytometry. Materials and Methods: The study included samples of 239 patients submitted to cytologic analysis and flow cytometry analysis of cellular markers. Statistica 7.1 software was employed on statistical analysis. Results: Study results showed cytologic puncture to be an appropriate method to obtain an adequate cell count for flow cytometry analysis irrespective of the benign or malignant nature of lymphoproliferative disease. Analysis of the percentage of lymphatic cells according to their size (cells of normal and small lymphocyte characteristics, SLY; cells of large lymphatic cell characteristics, LLY; and cells of other mononuclear cell characteristics, OMS) showed benign lymphoproliferative diseases to be characterized by SLY, B-non-Hodgkin's lymphomas mostly by LLY, and lymphomas of non-B-cell origin mostly by OMS. B-lymphocyte clonality is generally a characteristic of B-malignant lymphoproliferative diseases (B-NHL). In some cases of B-NHL, clonality cannot be demonstrated if the percentage of clonal lymphatic cells is low (in some cases of diffuse B large cell lymphoma abundant in T-lymphocytes), or if large lymphatic cells have been destroyed by manipulation (puncture, mononuclear separation, etc.). Conclusion: Cytodiagnosis is a simple and reliable method to differentiate benign from primary lymphoproliferative diseases (lymphomas) and secondary malignancies (metastases), as well as in typing and subtyping of malignant lymphomas. As cytologic puncture provides an adequate cell count for flow cytometry in lymphoproliferative disease, diagnostic accuracy and reliability are upgraded by combined use of these two methods. Background: Soft tissue myoepithelioma is an extremely rare neoplasm arising in the subcutaneous or deep subfascial soft tissues of the extremities. To our knowledge, the cytologic features of soft tissue myoepithelioma have been not described in the English literature. We report the imprint cytologic features of a case of soft tissue myoepithelioma. Case: A 18-year-old man presented with a solitary mass on the radial aspect of the right forearm for 5 months. MR images showed a well defined mass in the forearm. The mass was excised. The tumor measured 3.0 · 1.8 · 1.5 cm. The cut surface was well demarcated, lobulating, yellowish myxoid, and soft. The touch imprint cytologic smears from the mass were obtained. The alcohol-fixed smears were stained with hematoxylin-eosin and Papanicolaou stain. Microscopic examination showed round to ovalshaped tumor cells in single cells, sheets or clusters. These cells had relatively uniform, round to ovoid nuclei, with finely granular chromatin. The nuclei were centrally or eccentrically located. The cells had eosinophilic or indistinct cytoplasm. No significant nuclear atypia was observed. The background was myxoid. No necrosis was found. On immunohistochemical stain, the tumor cells were positive for cytokeratin (AE1/AE3) and S-100 protein. On the histologic examination, the tumor was multilobulated and myxoid. The tumor cells were round and epithelioid. There was no significant nuclear atypia and pleomorphism. On the immunohistochemical stain, tumor cells were positive for cytokeratin (AE1/AE3), CAM 5.2, S-100 protein, and calponin. Ultrastructurally, the cytoplasm contained thin myofilaments as well as scattered cell-to-cell attachments by desmosome. The appearances are consistent with soft tissue myoepithelioma, There was no evidence of recurrence ten months after surgery. Conclusion: An awareness of soft tissue myoepithelioma and its characteristic cytologic features and immunoreactivity with epithelial and myoepithelial markers is helpful in its correct diagnosis. Objective: An essentials part of all histological and cytological techniques is the preservation of cells and tissues. For immunocytochemistry (ICC) very important is preservation of antigens of the cells. There are various methods of fixation however each of them has advantages and disadvantages depend on results we want to achieve. In some cases available material is too small to make several smears each fixed in different way. Therefore it'd be good to have possibility to use the cell-smear second time. Aim of the Study: (1) select the best fixation method of the cellsmears for immunocytochemistry; (2) test if the second fixation in formalin of the cell-smears already processed (fixed in ethanol, stained H-E, decolorized) improve the quality of immunocytochemical staining?Material and Methods: We tested twenty cases of breast carcinoma. The cell-smears were fixed in six different ways and stained immunocytochemically (ICC) for estrogen receptor (ER), progesterone receptor (PR) and cytokeratin AE1/AE3. Results and Conclusion: The best ICC staining we obtained in cell-smears fixed in formalin. The alcohol fixation resulted in weaker reaction but application of formalin for decolorized cellsmears improve the quality of ICC.We recommend fixation in formalin for ICC. If you have to use second time cell-smears fixed in alcohol, stained H-E and decolorized we recommend second fixation in formalin before ICC. Aims: Subcutaneous masses of the abdominal wall are uncommon. However, a variety of benign and malignant entities can be found in this region. Materials and Methods: We report a series of 46 fine-needle aspirations (FNAs) of subcutaneous masses involving the abdominal wall, witch were diagnosed over a period of nine years (2000) (2001) (2002) (2003) (2004) (2005) (2006) (2007) (2008) . There were 12 males and 34 females with an age range 3-76 years. All FNAs were done by a cytopathologist using 22-25G needles, followed by staining with Pap and MGG. Subsequently the FNAC diagnosis were correlated with the histological diagnosis (n = 39). Results: Diagnosis was possible in 45 cases. In one case the material was insufficient for diagnosis. The FNAC diagnosis included 14 soft tissue lesions (lipoma, three; desmoid fibromatosis, seven; and fibrous tumors, four), metastatic tumors (carcinoma, six and lymphoma, four), 17 endometriosis, three inflammatory process and however many singly scattered spindle cells with the same nuclear features were also present. The cytoplasm was focally vacuolated. Necrotic debris was present in the background. Immunohistochemical studies performed on the cell block material confirmed the suspected morphologic impression of epithelioid angiosarcoma. In addition, subsequent core biopsies and autopsy material reconfirmed the cytologic diagnosis. Differential diagnosis included Hü rthle cell carcinoma, anaplastic thyroid carcinoma, medullary thyroid carcinoma and metastatic carcinoma to the thyroid. Conclusion: Epithelioid angiosarcoma of the thyroid gland is a rare and aggressive neoplasm that can be accurately diagnosed by FNA. Immunocytochemical studies, clinical correlation and consideration of differential diagnostic possibilities are crucial to arrive at the correct diagnosis. Introduction: The rhabdomyosarcoma is the most common paratesticular sarcoma in children and young adults. The alveolar subtype is rarely encountered, comprising 6-10% of all paratesticular rhabdomyosarcomas. Unlike the embryonal, the alveolar subtype is associated with an unfavourable prognosis, and therefore accurate diagnosis is vital. Case Report: A 19-year-old male presented with what appeared to be a testicular tumor approximately 5cm in diameter. Upon resection, it was discovered that the tumor in fact originated in the scrotum. Cytologic imprints for a rapid intraoperative diagnosis were obtained from tiny tissue samples of the tumor. The imprints were stained with Hemacolor and Papanicolaou stains.Total tumor resection and histologic examination followed. Cytologic Features: The cytologic examination revealed mediumsized round cells with eccentric nuclei and inconspicuous nucleoli forming rosette-like structures as well as enlarged dispersed cells with compact angular cytoplasm and pleiomorphic nuclei. The cytomorphologic examination excluded a primary testicular tumor. The diagnosis was of a malignant mesenchymal tumor. Histologic Diagnosis: The histologic diagnosis was of an alveolar rhabdomyosarcoma. The tumor stained positive for myoglobulin, Myo D1, SMA and desmin and negative for S-100, HCG, AFP, vimentin, CD117, PLAP, CD30, CAM 5-2, and pankeratin, confirming the initial diagnosis. Conclusion: Although rare, rhabdomyosarcoma should always be considered in the differential diagnosis of a paratesticular tumor, especially in young adults. The cytologic examination can be helpful in the diagnostic workup, particularly in excluding a primary testicular tumor and discriminating between a neoplastic and nonneoplastic lesion. Introduction: Although FNB has been well established in head and neck tumors in adults, it is not yet widely performed in children and adolescents.This study attempts to demonstrate the reliability, effectiveness and safety of this method in childhood. Materials and Methods: Our hospital is not a paediatric one, however, in a period of 6 years 76 children and adolescents, aged 6 months-16 years old, were referred to our cytology department with various head and neck tumors. In all cases FNB was performed by a cytologist, and the smears were stained with quick Giemsa (Hemacolor) and Papanicolaou stains. In 61 cases histologic examination followed, whereas in the remaining 15 the diagnosis was confirmed by patient follow-up. Results: The cytological examination revealed 61 benign and 15 malignant lesions:16 reactive lymph nodes, three cases of tuberculous lymphadenitis, seven branchial cysts, three lipomas, three neurilemmomas, three dermoid cysts, three common blue naevi, three parotid pleiomorphic adenomas, two benign fibrous histiocytomas, two benign fibrohistiocytic tumors, two benign parotid cysts, two lymphangiomas, one benign fibrous tumor, one nodular fasciitis, one hematoma, one eosinophilic granuloma, one intraparotid lipoma, one ganglial cyst, one meningioma, one juvenile xanthogranuloma, one neurinoma, one trichoepithelioma, one pilomatrixoma and one fibrolipoma, four embryonal rhabdomyosarcomas, three Hodgkin lymphomas, two dermatofibrosarcomas protuberans, two neuroblastomas, one Non-Hodgkin B-cell lymphoma, one lymph node infiltration from CLL, one malignant melanoma and one suspicious for liposarcoma. Correlation with available histologic examinations showed that there were no false-positive or false-negative diagnoses, whereas in eight cases there was discrepancy in neoplasm subtyping between cytology and histology. Overall concordance between cytologic and histologic diagnosis was 87%. Conclusion: In experienced hands, FNB of head and neck tumors in children and adolescents is comparable in accuracy to the FNB of head and neck tumors in adults. Therefore, it should be an essential part of the diagnostic workup in this age group as well. Objective: The purpose of this study was to assess the reproducibility of cytological diagnosis of colorectal polyps. Material and Methods: A total of 193 brushing specimens from colorectal polyps with corresponding histopathological diagnosis were reviewed. Thin Prep slides were evaluated twice by three cytopathologists with an interval of at least six months. Each reviewer was asked to categorize each slide into the following four categories: within normal limits, adenomas with low grade atypical lesions, adenomas with high grade atypical lesions and adenocarcinomas. Inadequate specimens were excluded from the study. Agreement was analysed using kappa statistics. Results: In the first round, there were difficulties in some cases concerning differential diagnosis of adenomas with high grade atypical lesions from adenocarcinomas. In the second round observer agreement was better, with kappa value between histopathological findings and cytological diagnoses 0.9. Total accuracy was 97.8%, sensitivity 98.13% and specificity 96.43%. Positive and negative predictive value was 99.37% and 90% respectively. Conclusions: The overall inter-and intraobserver agreement was good for brush cytology of colorectal polyps using Thin Prep method. Liquid based cytology appears to be highly accurate and reliable cytological method for diagnostic approach of colorectal polyps and could be added as a complementary technique to biopsy for the improvement of the diagnosis. Objective: Thyroid fine-needle aspiration (FNA) samples featuring a follicular patterned monotonous Hü rthle (oncocytic) cell population cannot reliably be diagnosed as either benign or malignant. Consequently, these lesions are eligible for surgical excision to determine their biological behavior. Most malignancy-related immunohistochemical markers proposed to reduce the number of unnecessary thyroidectomies lack sensitivity and specificity in the presence of Hü rthle cells. We recently identified cyclin D3 overexpression on histological sections of Hü rthle cell carcinomas. In the present study, we assessed the diagnostic value of cyclin D3 immunohistochemistry added to routine cytology. Material and Methods: Fifty-one FNAs suspicious for Hurtle cell neoplasia with histological follow-up (19 malignant cases) were examined. We evaluated the diagnostic value of cyclin D3 in cell block (CB) preparations and compared it to the closely related cyclin D1 protein.Results: As suggested by previous studies, the ‡25% positive cells was used as cut-off point. Cyclin D1 and D3 were highly specific (both 100%) and fairly accurate (75% versus 92%) in distinguishing benign from malignant oncocytic neoplasms, providing a strong positive predictive value (PPV; both 100%); however, their use yielded low sensibility (32% versus 79%) and low negative predictive values (NPV; 71% versus 89%). These latter parameters improved when samples positive for at least one of the cyclins were considered (sensibility: 89%; specificity: 100%; PPV: 100%; NPV: 94%; accuracy: 96%). Conversely, by adopting balanced ROC-derived positive cutoff values, cyclin D1 ( ‡6,5%) and D3 ( ‡7,5%) were highly sensible (both 100%) and accurate (90% versus 94%) in distinguishing benign from malignant oncocytic neoplasms, providing a strong NPV (both 100%); however, their yielded low specificity (84% versus 91%) and low PPV (79% versus 86%), which improved when samples positive for both cyclins were considered (sensibility: 100%; specificity: 94%; PPV: 90%; NPV: 100%; accuracy: 96%). Conclusions: Cyclin D3 increases the suspicion of malignancy in indeterminate oncocytic follicular lesions; its diagnostic performance depends on the positive cut-off point employed and is further enhanced when combined with cyclin D1. cell adenoma. In 4% (3 cases) of analyzed FNAB of parathyroid gland nodules adenoma was not properly recognized. In one case due to cystic degeneration of a nodule, in other case there were intranuclear inclusions present so it was mistaken for papillary thyroid carcinoma, and in third case it was reported as oncocytic thyroid adenoma. Conclusion: Ultrasound-guided FNAB, with data about localisation and ehosonographic structure of a nodule, assuming there is adequate material obtained, can be diagnostic for parathyroid gland adenoma in 96% of cases. Possible pitfalls are oncocytic type of parathyroid adenoma which can be mistaken for oncocytic thyroid adenoma, presence of intranuclear inclusions which can be mistaken for papillary thyroid carcinoma and cystic degeneration of a nodule because of epithelial cells absence. Mistakes can be avoided if we perform PTH level in FNAB obtained material. Introduction: Orbital tumors are relatively uncommon and the first clinical finding is unilateral exophtalmos except for some pediatric tumors, which are bilateral. Fine needle aspiration cytology (FNAC) is an useful technique for evaluation of orbital tumors, however, its use is infrequent because the proximity of the eye. The close collaboration between pathologist and ophthalmologist can avoid ocular complications. Materials and Methods: In all cases the FNAC were performed using 23 and 25-gauge needle attached to a syringe and handle in the usual manner. The aspirated material was fixed as air-dried smears for Diff-Quik staining or in alcohol for the Papanicolaou method. Some material was reserved for immunohistochemical and molecular studies. Results: Most of the patients were adults. Two cases were bilateral (Rosai-Dorfman disease and embryonal rhabdomyosarcoma). The malignant tumors were less frequent than benign lesions (28 cases of malignancies and 35 cases of benign conditions) with 15 cases of metastasic tumors from breast, lung and skin. The lymphoproliferative diseases constituted 11.47% of the serie with one case of follicular hyperplasia, five non-Hodgkin lymphoma including two low-grade lymphoma and one plasmacytoma. The 35 benign lesions included cases of fibrous solitary tumor, amiloid tumor, schwannoma, lymphangioma, fibrous histyocitoma and others. The only case of the rare Erdheim-Chester disease was diagnosed as a proliferation of histiocytic cells. In this serie there were no complications in no case. Conclusion: FNAC is a rapid, easily available technique for evaluation of orbital and ocular adnexa and it avoids aggressive procedures. The close collaboration between pathologists and ophthalmologists is a necessary condition for achieving representative material. Objective: The objective of this study was to evaluate the sensitivity and specificity of modified brush biopsy without computer-assisted analysis in the detection of oral premalignant and malignant lesions. Study Design: Ninety-four patients attending outpatient clinics who exhibited oral lesions suspicious of premalignancy or malignancy were enrolled. All patients underwent an oral brush biopsy using a baby toothbrush followed by a scalpel biopsy. The specimens were analyzed manually in a double-blinded fashion. Sensitivity and specificity were used for the statistical analysis of the samples. Statistical significance was determined using the normal approximation to the binomial distribution of matched results, approximated by the Student t distribution mean test (paired t test). Results: Seventy-nine patients with adequate transepithelial brush biopsy samples were included in the study group. When compared to scalpel biopsy, the statistical sensitivity of the brush biopsy was greater than 76.8% (P < .05) while the statistical specificity was greater than 93.3% (P < .05). There were 4 false negative brush biopsy cases. The 4 false negative patients turned out to be dysplasia/ malignancy on histopathology. All 4 were patients with clinical oral submucous fibrosis. Conclusion: The oral brush biopsy without computer-assisted analysis was found to be a painless, noninvasive test for evaluating oral lesions. The toothbrush brush biopsy with manual analysis had lower sensitivity and specificity than the commercially available oral brush biopsy with computer-assisted analysis. The results demonstrate that by using a toothbrush to obtain an oral brush biopsy sample, oral lesions can be easily evaluated in a resource challenged settings to rule out dysplasia and carcinoma. Objectives: Male factors are responsible for about half of all infertility cases. Until recently, testicular biopsy was the standard method to ascertain the aetiology of azoospermia. Fine needle aspiration cytology has gained increasing popularity as a simple and minimally invasive procedure that can help in assessing testicular function accurately. This study was aimed at addressing the question whether testicular fine needle aspiration (FNA) may be used as a first-line diagnostic modality in azoospermia and to assess its usefulness in the diagnostic protocol.

The FNA was performed in 78 consecutive azoospermic patients. To obviate sampling errors both testes were aspirated, except when contraindicated. Routine haematoxylin and eosin as well as Romanowsky staining was performed on the smears. Results: The smears were categorized on cytological examination into normal spermatogenesis in 35 (50%) patients, Sertoli cells only syndrome in 22 (31.4%) and maturation arrest at the spermatocyte/spermatid level was seen in 13 (18.4%) patients. There were eight (10.2%) cases with scant smears where cytological diagnosis could not be made. A good correlation between cytological smears and histological sections was found in 54 of 58 testes (93.1%) in which histopathological confirmation was available. Conclusions: Testicular FNA may be utilized as a first-line investigative modality in patients with azoospermia, provided the procedure is performed and interpreted by experts. Objectives: The application of image analysis system in order to measure morphological parameters on immunochemistry stains is an important procedure in the interpretation of breast cancer biopsies. The aim of this study is to investigate the influence of JPG Compression on these parameters. Materials and Methods: Seventy-five cases of breast FNA's have been analyzed by a non-medical person (Medical Physicist) using a custom image analysis system (Image Pro Plus) connected to a color digital camera. After the digitization of the slides and the nuclei segmentation, in the above cases 24 morphometrical parameters (area, optical density, perimeter e.t.c) as well as label index, quick score and heterogeneity concentration have been measured.The initial non compressed images were saved using the common compression algorithm JPEG, at several compression levels. The same parameters were measured for the original and the compressed images and the values have been added in a database in order to perform statistical analysis. Results: Measurements do not differ for various levels of compression. However, according to the compression level measured results were distinct, but a very good correlation can be observed among them (r > 0.85 and P < 0.01). Conclusions: The fact that common image compression does not affect significantly the morphological parameters indicates that a cytopathology department could easily use telemedicine applications in daily clinical practice since compressed images could be sent faster without loosing diagnostic information.right upper lung with irregular margins and a cystic lesion in the right thoracic wall that was expanded up to the pleura. Several smears were taken from the lesion. Both conventional and liquid based cytology methods were stained with Papanicolaou, May Gr} unwald Giemsa stain and microbiology conventional stains. Results: Smears screening revealed the presence of a neutrophilic clustering with a number of very small deep dark corpuscules that special laboratory studies revealed the presence of Sporothrix Schenckii. Conclusions: We wish to emphasize the contribution of FNA cytology as the first approach in the evaluation of sporotrichosis in parallel with the exclusion of malignancy and tuberculosis. Objective: Evaluation of cytologic features of distant/unusual presentations of lung carcinoma in two challenging cases. Materials and Methods: Cytologic material from two cases of lung cancers, one previously unknown, was examined in our institution. Patient 1 presented in an outpatient clinic with a 1.5 cm soft tissue nodule in the left peri-umbilical area. The nodule was sampled by fine needle aspiration, using a 23 gauge needle, 5/8 inch in length, attached to a 10 cc syringe, mounted on a Cameco Ò syringe holder. Air dried Diff-Quik Ó and alcohol-fixed Papanicolaou stained slides were examined. Patient 2 was a 48 year old woman with a history of widely disseminated lung carcinoma, who presented to her gynaecologist with inter-menstrual spotting. A cervico-vaginal sample processed for a thin-prep Ò slide and an endometrial biopsy were simultaneously obtained and routinely processed. Immunohistochemistry was employed to process case 2. Results: FNA evaluation of peri-umbilical nodule in patient 1 showed epithelial cells in variably sized three dimensional clusters and single cells with dense cytoplasm, intercellular bridges and pleomorphic and hyperchromatic nuclei. Some of these malignant cells were bizarre and overtly keratinized, allowing an unequivocal diagnosis of squamous cell carcinoma with lung likely being the primary site. Subsequent chest x-ray confirmed a left lung nodule, which at CT guided biopsy was confirmed as primary squamous cell carcinoma. Cervicovaginal cytology in the second patient showed three-dimensional small clusters of glandular cells. Cells were characterized by cytoplasmic vacuoles displacing the nuclei that were pleomorphic and hyperchromatic. Tumor diathesis or a bloody background was not evident. A diagnosis of adenocarcinoma, favour extrauterine, was rendered, pending further immunohistochemical evaluation. A concurrent endometrial biopsy reproduced the tumor cell features that were seen at cytology. Immunohistochemical evaluation with thyroid transcription factor-1 (TTF-1) (positive) and thyroglobulin (negative) confirmed this case to be lung adenocarcinoma metastatic to endometrium. Conclusion: Metastatic lung carcinoma can present in unusual locations including subcutaneous tissue and endometrium. When accessible, metastatic overt lesions may easily and safely be sampled with a fine needle to produce meaningful samples that can aid in triaging the case further and can cut down the treatment cost significantly. Very rarely, metastatic tumor may also be seen in cervicovaginal samples when endometrium is involved. In such a setting, awareness of patient history and clinical findings and the presence of tumor cells in the cervico-vaginal sample can alert for the possibility of a metastatic lesion. Tumor diathesis or old blood, features typical of an endometrial primary adenocarcinoma, are typically absent and immunohistochemical evaluation for TTF-1 can help in identifying lung likely to be the primary source. Metastatic squamous cell carcinoma from lung would look no different from primary cervical squamous cell carcinoma at cytology, but the former is much less likely to be positive for P16, a surrogate marker for high risk HPV, a ubiquitous etiologic causative agent for cervical squamous cell carcinoma. Introduction: Endobronchial ultrasound-guided fine-needle aspiration (EBUS-FNA) is an accurate and useful procedure for the diagnosis of thoracic lesions and mediastinal lymph node staging for lung cancer. Objective: To evaluate the non-neoplastic pathology diagnosed by means of EBUS-FNA.