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Gynecologic & Obstetrics 232A ANNUAL MEETING ABSTRACTS (28S, 18S, and 5.8S). To control for overall ploidy, 3 different single copy reference unanimously diagnosed by all 11 pathologists as being invasive. There was not a single genes were used for normalization (HBB, PTGS2, and STK19). Copy numbers were case where all 11 pathologists agreed that there was no invasion. In 32 cases (71%), the determined using the using the relative standard curve method. Genomic DNA was pathologists did not unanimously agree on the presence or absence of invasion. Mean isolated from frozen tissue sections of radical prostatectomy specimens from n = 21 agreement for depth of invasion was only moderate (mean κ =0.50), and ranged from patients (Gleason scores 6-9). Relative normalized target gene quantity in tumor DNA poor (κ= 0.12) to excellent (κ =0.92). was determined from the standard curves and divided by the relative normalized target Conclusions: There was only fair agreement among gynecologic pathologists in gene quantity of the calibrator, which consisted of DNA from each patient’s matched determining the presence of invasion in vulvar carcinoma. In cases where pathologists normal prostate tissue. agreed there was invasion, agreement on depth was only moderate. The fair to moderate Results: The median ratio of rDNA copies in tumor vs. normal was 1.45 for 5.8S, 1.43 agreement in this study may, in part, be due to the preferential selection of a larger for 18S, and 1.56 for 28S. The difference between these values and the expected tumor/ proportion of diagnostically challenging cases. This study highlights that assessing normal ratio of 1.0 (null hypothesis) was significant for each target (5.8S p=0.0055, whether or not there is invasion, and ascertaining the critical ≤ 1mm depth, still poses 18S p=0.0072, 28S p=0.0129; Wilcoxon Sign Rank test). Overall tumor/normal ratios a diagnostic challenge for pathologists. were increased in 16 of the 21 cases. There was a high level of concordance in median relative copy number (tumor/normal) using different reference genes (e.g. HBB, PTGS2, 1037 Endometrial Adenosarcomas: Diagnostic Use of Ki-67 Proliferation RP1) with the same target (e.g. 18S rDNA), and using the different target genes in the Marker as an Adjunct to Morphologic Diagnosis locus (e.g. for a given patient if 5.8S rDNA was amplified then the 18S and 28S genes N Aggarwal, R Bhargava, E Elishaev. University of Pittsburgh School of Medicine, were also amplified). Pittsburgh, PA. Conclusions: In the majority of cases of prostate cancer there was an increase in Background: Endometrial adenosarcomas (AS) are rare biphasic neoplasms of the copy number of the entire rDNA locus (relative increase ∼1.5 fold). Since there are female genital tract containing benign glands and a low grade sarcomatous mesenchymal approximately 400 copies of this repeat unit in normal cells, these increases are on the component. Diagnostic criteria include increased stromal cellularity in periglandular order of hundreds of extra copies. These results raise the possibility that rDNA gene distribution (periglandular stromal cuffing) accompanied by variable degrees of cytologic copy number may: i) influence nucleolar size expansion in cancer cells, and/or ii) play atypia and mitotic activity. When not all the criteria are fulfilled a diagnosis of benign an important role in the development and/or progression prostate cancer. lesions are given and AS is considered in retrospect upon recurrence of the tumor. Design: Eight cases of AS were identified at Magee-Womens Hospital between 2004 1035 The Study of Xp11 Translocation Renal Cell Carcinoma (RCC) in and 2008 and were compared to 15 cases of endometrial polyps (EP) and 14 cases of Adults by TMA, IHC and FISH atypical polypoid adenomyomas (APA). Pertinent 1-3 blocks/case were selected and M Zhong, L Osborne, MJ Merino, M Hameed, S Aisner. University of Medicine and stained for Ki-67 proliferation marker, caldesmon, smooth muscle actin (SMA), desmin, Dentistry of New Jersey-New Jersey Medical School, Newark; National Cancer Institute, and CD10 with appropriate negative and positive controls. Bethesda; Memorial Sloan-Kettering Cancer Center, New York. Results: All cases of AS had a polypoid growth pattern with round or slit-like glands Background: Xp11 translocation renal cell carcinomas (RCC), a distinctive entity in with or without papillary projections into gland lumen. Increased periglandular stromal the 2004 WHO renal tumor classification, are rare neoplasms and are often encountered cellularity was observed focally or diffusely in all cases (8/8) of adenosarcoma, which in the pediatric and young adult population. The incidence is probably underestimated also demonstrated variable degrees of stromal nuclear atypia. The mitotic activity ranged partly due to unavailable genetic studies and overlapping histopathological morphology from 1/10 high power fields (HPF) to 15/10HPF. All cases of AS demonstrated distinct with clear cell and papillary RCC. TFE3 immunohistochemical assay has been used for increase in Ki-67 positive nuclei in the peri-glandular zone compared to adjacent stroma, the diagnosis of the Xp11.2 translocation RCC. However, for a definitive diagnosis, one regardless of the mitotic count. The average Ki-67 labeling index in peri-glandular zones has to rely on identifying the translocation by genetic and molecular studies. of AS was 20 % compared to less than 5 % in adjacent stroma. This zonation was not Design: A total of 120 consecutive adult (>18Y) RCC patient specimens (FFPE during observed in any case of APA or EP all of which showed scattered positive cells with the period 2001-2008) from our institute, were collected to construct a tissue micro less than 5% Ki 67 proliferation index. The AS stroma was positive for CD10, with array(TMA). IHC was performed on TMA using TFE3 antibody. The 2nd TMA was variable focal positive staining for SMA and desmin, and was negative for caldesmon. constructed from all the TFE3 positive specimens, 5 TFE3 negative and 5 TFE3 weak APA showed positivity for all muscle markers with SMA being the most consistent and positive specimens. A dual-color, break-apart FISH assay, which can detect the gene diffuse. EPs were negative for caldesmon, variably positive for desmin and SMA and rearrangement at Xp11 region, was applied to the 2nd TMA. diffusely positive for CD10. Results: Among the 120 RCC, 11(9.2%) cases were TFE3 positive . Among these 11 Conclusions: Distinct increase in peri-glandular Ki-67 staining pattern is very helpful cases, FISH assay showed split signal, which confirmed chromosome translocations and Ki 67 should be considered as an adjunct to the routine morphologic diagnosis of involving TFE3, in 5(4.2%) cases; fusion signal in 1 case; non-conclusive result in the AS. This could be especially useful in curettage specimens and other challenging lesions rest 5 cases(most likely due to the ages of the blocks); no split signal in all tested TFE3 that lack some of the classic criteria of an AS. negative and weak positive cases. Further more, no TFE3 fusion transcripts (ASPL, PRCC, CLTC, PSF and Nono), but TFE3 wide type were detected by RT-PCR in the 1038 New Antibodies for Clear Cell Carcinoma (Pax-8, HNF-α, vHL) Are tested case with weak TFE3 nuclear stain. Also Positive in Arias-Stella Reaction Conclusions: 1) Xp11.2 translocation RCC is not an uncommon neoplasm. The CE Aguilar, E Silva. Cedars-Sinai Medical Center, Los Angeles, CA. incidence in this group of 120 RCC patients is at least 4.2% in adults (confirmed Background: Recent studies have shown clear cell carcinoma of müllerian origin to by FISH). 2) TFE3 IHC is a relatively sensitive and specific assay for the diagnosis be positive for Pax-8, hepatocyte nuclear factor- α (HNF-α), and von Hippel Lindau Xp11.2 translocation RCC. The strong nuclear TFE3 stain is most likely indicative of (vHL). Clear cell carcinoma of müllerian origin can present a diagnostic challenge in Xp11.2 translocation. The weaker nuclear stain appears to be due to expression of full endometrial specimens showing areas of Arias-Stella. Therefore, Pax-8, HNF-α, vHL length TFE3 protein, rather than chimeric fusion protein due to translocation. 3) FISH were applied to products of conception showing areas of Arias-Stella reaction. assay is the most reliable method to detect Xp11 translocation RCC, when only FFPE Design: Ten products of conception specimens showing Arias-Stella reaction and ten material is available. cases of clear cell carcinoma were stained with Pax-8, HNF-α, vHL. Results: In all ten cases, Arias-Stella cells and secretory glands displayed Pax-8 and HNF-α nuclear positivity and vHL cytoplasmic staining. All three immunostains were Gynecologic & Obstetrics positive in each case of clear cell carcinoma with the expected pattern of staining. Conclusions: Clear cell carcinoma antibodies, Pax-8, HNF-α, and vHL, are positive 1036 Inter-Observer Agreement among Pathologists for Assessing in Arias-Stella reaction. These antibodies therefore cannot be applied to endometrial Invasion in Early Vulvar Squamous Cell Carcinoma: Still a Diagnostic specimens in which the differential diagnosis includes clear cell carcinoma and Arias- Challenge Stella reaction. Future immunohistochemical studies to determine whether other A Abdel-Mesih, D Daya, K Onuma, N Akhtar-Danesh, O Boutross-Tadross, K Ceballos, molecular targets involved in the clear cell carcinoma pathway(s) can also be detected W Chapman, T Colgan, P Deb, MR Nucci, E Oliva, M Sur, S Tang, A Lytwyn. McMaster in Arias-Stella reaction would be useful in this unusual but challenging and important University, Hamilton, ON, Canada; University of British Columbia, Vancouver, BC, distinction. Canada; University of Toronto, Toronto, ON, Canada; Harvard University, Boston, MA. 1039 Prognostic Factors of Adenocarcinoma of the Endocervix: Background: Despite published criteria, measuring depth of invasion to identify Pattern of Invasion vs Depth of Invasion early squamous cell carcinoma of the vulva remains difficult, yet critical for surgical I Aguilera-Barrantes, EG Silva.
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