Absence Of Staining Research Articles

S1647 A Surprising Diagnosis of Lynch Syndrome at Age 86

INTRODUCTION: Lynch syndrome (LS) is the most common inherited colorectal cancer (CRC) syndrome and accounts for up to 6% of all CRCs in the United States. The lifetime risk of CRC is near 75% for individuals with this inheritable highly expressed gene making identification of individuals and at-risk family members important for cancer prevention and management. CASE DESCRIPTION/METHODS: 86-year-old male in excellent physical condition with history of right-sided poorly differentiated invasive stage IIa adenocarcinoma status post right hemicolectomy 12 years ago presented after 6 episodes of painless hematochezia (hemoglobin 11.9 g/dL). He chose not to undergo chemotherapy and never had a follow-up colonoscopy following his initial CRC. He reported a family history of 1 sister who recovered from breast cancer and 2 deceased younger brothers who passed from colon cancer after his original diagnosis of CRC. Both parents experienced early death from cardiovascular disease. He had 6 children. Colonoscopy revealed normal ileocolonic anastomosis but with a 3 cm ulcerated, friable mass in transverse colon, a 2.5 cm mass in rectosigmoid colon, and a large, fungating 4cm ulcerated mass 5 cm from the anal verge. CT chest/abdomen/pelvis scan demonstrated extension of the transverse colonic mass through the colon wall into the mesentery with extensive nodal and hepatic metastatic disease. Pathology revealed invasive moderately differentiated adenocarcinoma positive for MLH1, MHS2, and PMS2 suggestive of Lynch syndrome; absent MSH6 staining was noted. Family members were recommended for genetic testing. The patient ultimately opted against treatment. DISCUSSION: The average age of colon cancer diagnosis for those with LS is 44 as compared to 68 for men without the genetic syndrome. This case demonstrates a particularly unique diagnosis of Lynch syndrome in an elderly male that was not diagnosed until his second bout of CRC. Of note, both cancers occurred after age 70. Strict adherence to the revised Bethesda guidelines can miss 68% of high microsatellite instability CRCs suggesting these guidelines are inadequate for LS screening, particularly if a full multigenerational family cancer history is not available. Multiple studies have demonstrated economic feasibility and advocate for universal LS testing of CRC tumors. This case supports such a paradigm. For identifying those who may be at risk for LS, we would prefer a predictive model without an overreliance on age to determine those appropriate for screening.Figure 1.: Endoscopic view of the 3 cm transverse colon (A), 2.5 cm rectosigmoid colon (B), and 4 cm anal verge (C) masses in an 86-year-old male discovered to have Lynch syndrome.Figure 2.: CT abdomen/pelvis showing transverse colon carcinoma with direct extension through the colon wall (red arrow) and into the mesentery associated with extensive nodal, as well as hepatic (yellow arrows) metastatic disease in an 86-year-old male discovered to have Lynch syndrome.

Downregulation of microRNA‑126 is inversely correlated with insulin receptor substrate‑1 protein expression in colorectal cancer and is associated with advanced stages of disease

Colorectal cancer (CRC) is a common human malignant tumor, and the fourth most common cause of cancer-associated mortality in China. However, the pathogenesis of CRC is not yet fully understood. The present study aimed to investigate the expression and clinical significance of microRNA (miR)-126 and insulin receptor substrate-1 (IRS-1), as well as the role of miR-126 in the prognosis of patients with CRC. A total of 86 colorectal tissue specimens, including 40 CRC and adjacent normal tissue, 26 colorectal adenoma tissue and 20 normal colorectal tissue samples, were collected for the present study. Reverse transcription-quantitative PCR analysis was performed to determine miR-126 and IRS-1 mRNA expression levels, while western blotting and immunohistochemistry (IHC) analyses were performed to determine IRS-1 protein expression levels. The correlation between miR-126 and IRS-1 expression, as well as the association between altered miR-126 and IRS-1 expression levels and clinicopathological characteristics, and the overall survival time of patients with CRC were assessed. The results demonstrated that miR-126 expression was significantly downregulated, while IRS-1 protein expression was upregulated in CRC tissues compared with that in adjacent normal tissues, colorectal adenoma tissues and normal colorectal tissues, respectively. IHC analysis exhibited strong positive staining of IRS-1 protein in CRC tissues, while absent or weak staining of IRS-1 protein was detected in adjacent normal tissues, colorectal adenoma tissues and normal colorectal tissues. miR-126 expression was inversely correlated with IRS-1 protein expression in CRC tissues (r=−0.420; P<0.05). Furthermore, downregulated miR-126 expression was associated with advanced clinicopathological characteristics of the disease and a shorter overall survival time in patients with CRC. Taken together, the results of the present study suggest that miR-126 downregulation may be a candidate molecular marker predictive of poor prognosis of patients with CRC.

Major histocompatibility complex class 1 (MHC1) loss among patients with glioblastoma (GBM).

e14523 Background: Immune evasion represents a hallmark behavior of cancer and may occur through many mechanisms. Among these, intact tumor antigen presentation at the cell surface is a fundamental prerequisite to achieving successful adaptive immunotherapy. The loss of MHC1 expression due to molecular events, including mutation, deletion, or epigenetic silencing of B2M is commonly acquired during immunotherapy. On the other hand, molecular events affecting antigen presentation machinery may be present prior to immunotherapy administration. Among these, TP53 mutations causing loss of ERAP1 and TAP1 expression compromise transport of MHC1 molecules from the endoplasmic reticulum to the cell surface and mutant peptide integration into the HLA context resulting in absent antigen presentation. Thus far, clinical trials of PD-1/L1 agents have failed to demonstrate a benefit for GBM patients and responses are seen only among a minority. Hence, we set out to assess the integrity of MHC1 expression by using immunohistochemistry. Methods: Immunohistochemical (IHC) stains for HLA, B and C were developed and validated with internal controls. Staining intensity and location (membrane-bound or cytoplasmic) was evaluated semi-quantitatively. The first 10 consecutive patients with GBM who were referred for neoepitope vaccine were evaluated. Results: Absent staining was seen among 6/10, negligible, or faint staining was present in 2, and only 2 tumors demonstrated intact membrane-bound expression. Conclusions: In addition to low tumor mutation burden and an immunosuppressive tumor microenvironment, MHC1 loss is a frequent event among patients with GBM, and may be a dominant cause of immunotherapy failure for as many as 80% of patients. Thus, development of strategies to reverse this loss may be an essential component of successful adaptive immunotherapy for this disease. These data suggest that routine assessment of MHC1 should become a component of eligibility checking for GBM patients being considered for an MHC-restricted approaches. [Table: see text]

The Role of immunohistochemical Expression of monoclonal VE1 antibody in detecting the BRAF gene mutation in patients with colorectal carcinoma.

Purposes of study : to evaluate clinical parameters and histopathological features of Colorectal carcinoma (CRC) and its association with BRAF mutation as immunohistochemical markers in Babylon province. Methods: This research is prospective study. The total patients are 42 Manual IHC staining procedure were done. Staining intensity were scored as: 0 ( negative ), 1 (weak diffuse staining in comparing to background staining ), 2 (moderate diffuse staining), and 3 (strong diffuse staining) . IHC scoring was regarding as positive when there was diffuse, homogenous and more than 80% of cytoplasmic staining area of cancer cells. Negative cases when there were absent staining or nuclear staining or weak isolated staining cells. Score 0 and 1 were consider negative and score 2 and 3 were positive. Results: Positive IHC staining of BRAF mutation was presenting more in older age group (65 ±15.23years) , male gender (60%) and left sided colon (60%) but there were insignificant association of these above parameters. Grade 2 and grade 3 of CRC was the highest frequency of positive BRAF cases (40%). Positive IHC staining for BRAF mutation expression is more frequency in T3, N1-2, and M0 stage, and stage III that shows 60%, 80%, 100%, 80% respectively. There were no significant association between BRAF IHC with TMN staging and grading systems. Conclusion: the current study found to be predominant in older age ( &gt; 65 years old) , high grade (G2-3) and high stage (III). Parameters of high grade &amp; stage associated with poor prognosis &amp; high mortality outcome. BRAF gene IHC expression could be consider an independent bad prognostic factor for patients with CRC .

Expression and Immunolocalization of Xenobiotic Transporters at the Human Blood‐Testis Barrier

The blood‐testis barrier (BTB) is formed by basal tight junctions between adjacent Sertoli cells of the seminiferous tubules and functions as a physical barrier to protect developing germ cells in the adluminal compartment from reproductive toxicants. Multiple xenobiotics are known to cross into the male genital tract (MGT) including male contraceptives, cancer chemotherapeutics, and environmental toxicants, although the mechanism(s) for their movement into the MGT is(are) largely unknown. Previous studies showed that several ATP‐binding cassette (ABC) and solute carrier (SLC) superfamily transporters are expressed in the testes and could potentially facilitate the disposition of xenobiotics into the MGT through concurrent uptake and efflux mechanisms. This study aimed to determine the expression and localization of xenobiotic transporters that may facilitate transepithelial transport across the human BTB. We used immunofluorescence staining to determine the cellular localization of these transporters in the several cell types found in human testes. OAT1, OAT2, OCTN1, and MRP3 were primarily localized to the basal membrane of Sertoli cells, whereas OCTN2, MRP6, and MRP7 were localized to both the basal membrane of Sertoli cells and the peritubular myoid cells that surround the basal membrane of Sertoli cells. Interestingly, CNT2 was localized to the lateral membranes of Sertoli cells and the peritubular myoid cells. OCT1, OCT2, and OCT3 were localized to the peritubular myoid cells and the Leydig cells, although there was moderate staining of spermatogenic cell membranes. However, definitive staining of CNT1, OAT3, MATE1, MATE2, OATP1A2, OATP1B1, OATP1B3, OATP1C1, OATP2A1, OATP2B1, OATP3A1, OATP4A1, and OATP6A1 was not observed due to non‐specific antibody staining or an absence of staining. The localization of CNT2, OAT1, OAT2, OCT1, OCT2, OCT3, OCTN1, OCTN2, MRP3, MRP6, and MRP7 suggests these transporters may contribute to the mechanism(s) by which xenobiotics can cross the BTB. Taken together, the localization of these pharmacologically relevant transporters provides insight into the selectivity of drug disposition across the BTB and may be useful in developing tools to understand and overcome the pharmacokinetic and pharmacodynamic difficulties presented by the BTB.Support or Funding Information5R01GM123643‐045R01GM129777‐02P30CA023074

Variable Expression of S100 Protein in Sinonasal Malignant Mucosal Melanoma: A Potential Diagnostic Pitfall.

Sinonasal malignant mucosal melanoma (SNM) is a rare, aggressive malignancy. The diagnosis of SNM is often quite challenging due to anatomical limitations, frequent lack of pigmentation, variable histologic appearances, and aberrant differentiation (e.g., positivity for cytokeratin, desmin, or neuroendocrine markers). S100 protein is routinely used as a standard screening marker for SNM, but it may lack optimal sensitivity. Our objective was to study the extent of immunohistochemical expression of S100 protein in SNM, and determine its diagnostic value by comparing it to a newer melanoma marker, SOX10. Twenty-three cases of sinonasal MMM were retrieved from the archival files of the Department of Pathology at UT Southwestern Medical Center. The patients included 14 men and 9 women, and ranged from 36 to 90years (mean 64.9years). Sections from blocks of formalin-fixed, paraffin-embedded tissue were used for immunohistochemical analysis with S100 protein and SOX10. The extent and intensity of immunostaining was recorded, and H-score was calculated. For a subset of negative or focally positive cases, S100 protein was repeated at a high-volume reference laboratory. S100 protein immunoexpression was quite variable in the SNM cases, with H-scores ranging from 0 to 300 (mean 123). While 11 of 23 cases exhibited strong and diffuse staining (H-score > 100) as expected for melanoma, 7 were weak and/or focal (H-score 1-100), and 5 were completely S100 protein-negative. For 10 cases, the negative or focal results were confirmed by reference laboratory staining. In contrast, all 23 SNM cases were diffusely and strongly positive for SOX10 (H-scores 210-300, mean 296). Our study demonstrated that S100 protein immunoexpression is extremely variable in SNM. Weak or even absent S100 protein staining is not uncommon in SNM, and should not dissuade pathologists from that diagnosis. Our data demonstrates that S100 protein is insufficiently sensitive to be used as a screening marker for SNM, but that SOX10 is consistently and robustly positive, and should therefore replace S100 protein for that purpose. Indeed, for any high-grade sinonasal tumor, pathologists must have a low threshold for utilizing additional markers to exclude the possibility of SNM.

P53 Immunohistochemical patterns in HPV-related neoplasms of the female lower genital tract can be mistaken for TP53 null or missense mutational patterns.

We have recently encountered p53 immunohistochemical (IHC) patterns in human papillomavirus (HPV)-associated carcinomas of the gynecologic tract, which were confused with absent (null) or overexpression TP53 mutational staining. We therefore evaluated p53 and p16 IHC in 25 squamous cell carcinomas (SCC) (16 vulva, 4 Bartholin's gland, and 5 cervix), 20 endocervical adenocarcinomas (EDAC), 14 high-grade squamous intraepithelial lesions (HSIL), 2 adenocarcinoma in situ (AIS), all of which exhibited morphologic features of HPV. Only cases showing diffuse/strong block-like p16 staining were included for further study. All EDACs underwent TP53 sequencing and HPV in situ hybridization (ISH) was performed in selected cases. p53 IHC staining fell into two main patterns. The most common was designated as "markedly reduced (null-like)" (absence or significantly attenuated staining in >70% of cells), which could be confused with true null mutational pattern. This was present in 14/25 (56%) SCCs, 7/14 (50%) HSILs, and 18/20 (90%) EDACs. The second notable pattern was "mid-epithelial (basal sparing)" (distinct absence of staining in basal cells juxtaposed with strong staining in parabasal cells), seen in 10/25 (40%) SCC, 7/14 (50%) HSIL, and none of the EDACs. There was scattered weak to moderate p53 staining (conventional wild type) in 1/25 (4%) SCC and 2/20 (10%) EDAC. No cases showed strong/diffuse overexpression. One EDAC had a TP53 missense mutation and exhibited "markedly reduced (null-like)" staining. HPV ISH revealed an inverse relationship with p53, cells positive for HPV mRNA were negative for p53. Knowledge of these patterns can help pathologists avoid misinterpreting p53 status in the setting of HPVA cancers.

Mucus characterisation in the Octopus vulgaris skin throughout its life cycle.

The development of the epidermis of octopus, Octopus vulgaris, throughout its life cycle was studied by conventional staining and histochemical techniques using lectins. The mantle, the arm and the two parts of the suckers: the infundibulum and the acetabulum were analysed independently. With the exception of the suckers, the general morphology of the epidermis does not vary from the first days post-hatching to adulthood. In general terms, histochemical techniques do not indicate changes in the composition of glycoconjugates of the epidermis main cells, epithelial and secretory cells. The epithelial cells of the mantle and arm show positivity for mannose (ConA+) in their apical portions, indicating the presence of n-glycoproteins that, among other things, provide lubrication to the surface of the body. In the suckers, the apical surface of the infundibulum contains sulphated glycosaminoglycans of the N-acetylglucosamine type that provide adhesive properties. In addition to observing three types of mucocytes, m1 and m2 are characteristic of the mantle and arm, and m3 is found in the suckers. The paralarva epidermis is characterised by the presence of Kölliker's organs whose exact function is unknown. In this study, the absence of staining with alcian blue/periodic acid-Schiff(AB/PAS) prevents the possibility of attributing a secretory function. Nevertheless, the linkage of three lectins (WGA, LEL and GSL-I) in the fascicle of the organ suggests the presence of proteoglycans rich in N-acetylglucosamine that would mainly have a structural role.

Major p53 immunohistochemical patterns in in situ and invasive squamous cell carcinomas of the vulva and correlation with TP53 mutation status.

The recent literature has shown that vulvar squamous cell carcinoma (VSCC) can be stratified into two prognostically relevant groups based on human papillomavirus (HPV) status. The prognostic value of p53 for further sub-stratification, particularly in the HPV-independent group, has not been agreed upon. This disagreement is likely due to tremendous variations in p53 immunohistochemical (IHC) interpretation. To address this problem, we sought to compare p53 IHC patterns with TP53 mutation status. We studied 61 VSCC (48 conventional VSCC, 2 VSCC with sarcomatoid features, and 11 verrucous carcinomas) and 42 in situ lesions (30 differentiated vulvar intraepithelial neoplasia [dVIN], 9 differentiated exophytic vulvar intraepithelial lesions [deVIL], and 3 high-grade squamous intraepithelial lesions or usual vulvar intraepithelial neoplasia [HSIL/uVIN]). IHC for p16 and p53, and sequencing of TP53 exons 4-9 were performed. HPV in situ hybridization (ISH) was performed in selected cases. We identified six major p53 IHC patterns, two wild-type patterns: (1) scattered, (2) mid-epithelial expression (with basal sparing), and four mutant patterns: (3) basal overexpression, (4) parabasal/diffuse overexpression, (5) absent, and (6) cytoplasmic expression. These IHC patterns were consistent with TP53 mutation status in 58/61 (95%) VSCC and 39/42 (93%) in situ lesions. Cases that exhibited strong scattered staining and those with a weak basal overexpression pattern could be easily confused. The mid-epithelial pattern was exclusively observed in p16-positive lesions; the basal and parabasal layers that had absent p53 staining, appeared to correlate with the cells that were positive for HPV-ISH. This study describes a pattern-based p53 IHC interpretation framework, which can be utilized as a surrogate marker for TP53 mutational status in both VSCC and vulvar in situ lesions.

Prognostic and Predictive Role of CXC Chemokine Receptor 4 in Metastatic Colorectal Cancer Patients.

Colorectal cancer (CRC) is the third leading cause of cancer-related death worldwide. About 30% of patients present with metastatic disease involving predominantly the liver and a similar percentage will develop distant metastases later after removal of the primary tumor. In metastatic CRC, chemotherapies and biological drugs have prolonged survival for up to 30 months. However, there is a great need for biomarkers predictive of response and prognosis to optimize treatments. CXC chemokine receptor 4 (CXCR4) is a chemokine receptor; it binds to CXCL12 and plays a central role in colon cancer cells' growth and dissemination. CXCR4 was evaluated in CRC primary tissues by immunohistochemistry. Formalin-fixed, paraffin-embedded 4-μm tissue sections were immunostained using a biotin-streptavidin-peroxidase method and categorized into 2 semiquantitative classes: (i) absence of staining, ≤50% positive cells (negative/low) and (ii) >50% positive cells (high). Associations between clinic-pathologic variables and CXCR4 expression were evaluated using the χ test. The Kaplan-Meier product-limit method was applied to graph overall survival (OS). OS was defined as the time elapsed from diagnosis to death from any cause. Univariate analysis was carried out using the log-rank test. Cox proportional hazards regression was used to analyze the effect of several risk factors on OS. Seventy-eight primary adenocarcinomas were analyzed; 26 were categorized as negative/low and 52 as high. Age, sex, performance status, site of metastases, KRAS mutational status, type of first-line therapy, and a number of therapy lines did not correlate with CXCR4 expression. Although not significant (P=0.0533), high CXCR4 expression was more frequently localized on the right side of the colon. Significant correlations were detected with grading (P=0.0041) and response to first-line anti-epidermal growth factor receptors agents (P<0.0001), bevacizumab (P=0.0029), and chemotherapy alone (P=0.0260). At a median follow-up of 53 months, 77 deaths have been registered. Grading [hazard ratio (HR): 1.42; confidence interval (CI): 0.89-2.28; P<0.0001], KRAS mutational status (HR: 1.73; CI: 1.03-290; P=0.0133), response to first-line chemotherapy (HR: 3.39; CI: 2.10-5.48; P<0.0001), and CXCR4 expression (HR: 3.18; CI: 2.01-5.02; P<0.0001) showed prognostic power at univariate and multivariate analyses. In the present report, we show that CXCR4 expression on the primary tumor is an independent prognostic factor and correlates with response to first-line chemotherapy in metastatic CRC patients.

Hot spots in fetal human penile and clitoral development

To better understand how the human fetal penis and clitoris grows and remodels, we undertook an investigation to define active areas of cellular proliferation and programmed cell death spatially and temporally during development of human fetal external genitalia from the indifferent stage (8 weeks) to 18 weeks of gestation. Fifty normal human fetal penile and clitoral specimens were examined using macroscopic imaging, scanning electron microscopy and immunohistochemical localization for the cellular proliferation and apoptotic markers, Ki67 and Caspase-3, respectively. A number of hot spots of cellular proliferation characterized by Ki67 localization are present in the penis and clitoris especially early in development, most notably in the corporal body, glans, remodeling glanular urethra, the urethral plate, the roof of the urethral groove and the fully formed penile urethra. The 12-fold increase in penile length over 10 weeks of growth from 8 to 18 weeks of gestation based on Ki67 labelling appears to be driven by cellular proliferation in the corporal body and glans. Throughout all ages in both the developing penis and clitoris Ki67 labeling was consistently elevated in the ventral epidermis and ventral mesenchyme relative to the dorsal counterparts. This finding is consistent with the intense morphogenetic activity/remodeling in the ventral half of the genital tubercle in both sexes involving formation of the urethral/vestibular plates, canalization of the urethral/vestibular plates and fusion of the urethral folds to form the penile urethra. Areas of reduced or absent Ki67 staining include the urethral fold epithelium that fuses to form the penile tubular urethra. In contrast, the urethral fold mesenchyme is positive for Ki67. Apoptosis was rarely noted in the developing penis and clitoris; the only area of minimal Caspase-3 localization was in the epithelium of the ventral epithelial glanular channel remodeling.

Heterogeneity of p16 immunohistochemistry and increased sensitivity of RNA in situ hybridization in cytology specimens of HPV-related head and neck squamous cell carcinoma.

Many patients with human papillomavirus (HPV)-related head and neck squamous cell carcinoma (HPV-HNSCC) initially present with cervical lymph node metastases. Although p16 immunohistochemistry (IHC) is the most commonly used surrogate marker for HPV, however criteria in cytologic material are not well established. The objective of this study was to better characterize p16 IHC in cell blocks of metastatic HPV-HNSCC, and to evaluate the performance of HPV RNA in situ hybridization (RNA ISH). p16 IHC was performed on cell blocks from 97 metastatic HPV-HNSCC fine-needle aspiration specimens with HPV status confirmed by DNA or RNAISH or polymerase chain reaction (PCR). Tumor cellularity (<100 cells, 100-500 cells, and >500 cells) and quality (presence of cell clusters, necrosis) were recorded. p16 staining intensity and extent (1%-9%, 10%-69%, and ≥70%) were scored. In addition, RNA ISH was performed on 38 PCR-positive cases. p16 IHC was positive in 90 of 97 cases (93%), demonstrating variable patterns. p16 staining was found to be moderate to strong in 69 cases, with 37 cases (38%) demonstrating positivity in ≥70% of tumor cells. Weak staining occurred in 21 cases (22%) and 7 cases (7%) were negative. Of the 60 cases with weak and/or absent expression or staining in <70% of cells, 30 cases (50%) had <100 tumor cells, 12 (20%) lacked cell clusters, and 19 cases (32%) had extensive necrosis. RNA ISH was positive in 37 of 38 cases (97%) that were HPV positive by PCR. p16 is heterogeneous in cell blocks of metastatic HPV-HNSCC, suggesting that any p16 positivity should prompt confirmatory HPV studies. RNA ISH appears to demonstrate high sensitivity, and laboratories even may consider using RNA ISH as a first-line HPV test in cytologic specimens.

Solid Variant of Serous Cystadenoma of the Pancreas, Cytology Findings

Abstract Introduction Serous cystadenoma of the pancreas is the most common benign neoplasm of the pancreas, tending to have a very good prognosis and treatable by surgical excision. The solid variant of serous cystadenoma of the pancreas is a very rare subtype with only a few cases reported in recent literature, mostly in elderly females. Case History An asymptomatic 85-year-old woman presented with a large pancreatic mass. Endoscopic ultrasound findings were consistent with an ill-defined large isoechoic mass involving almost the entire pancreas, with no significant common bile duct dilation. Fine-needle aspiration was performed. Results The smears showed groups of cells with uniform nuclei, ample cytoplasm, inconspicuous nucleoli, minimal cytologic atypia, and absent mitotic activity, suggesting a benign or low-grade lesion. A solid aggregate of these cells was present in the cell block, which was positive for CK7, inhibin, and PAS. Negative staining for chromogranin, synaptophysin, and B-catenin helped exclude neuroendocrine tumors and solid pseudopapillary neoplasm. Additionally, absent staining for PAX-8, ER, PR, BRST-2, GATA-3, CK20, CDX2, Melan-A, and calretinin helped exclude other metastatic clear cell lesions. Conclusion Diagnosis of solid variant serous cystadenoma of the pancreas can be challenging due to nonspecific clinical symptoms and misdiagnosis as other solid pancreatic tumors. Based on the clinical, radiologic, cytologic, and immunohistochemical features, we favor a diagnosis of solid variant of serous cystadenoma. Recognition of this lesion is important because the vast majority of solid tumors in the pancreas are malignant. The differential diagnosis includes the rare primary clear-cell “sugar” tumor of the pancreas, clear cell carcinoma, clear cell islet cell tumor, and metastatic renal cell carcinoma. Therefore, the diagnosis of solid-type serous cystadenoma of the pancreas is frequently dependent on a series of immunohistochemical and molecular studies.

Absence of Iodine Staining Associates With Progression of Esophageal Lesions in a Prospective Endoscopic Surveillance Study in China

Chromoendoscopy with iodine staining is used to identify esophageal squamous dysplasia and esophageal squamous cell carcinomas (ESCCs)-absence of staining indicates suspicious regions of dysplasia. However, screening detects precancerous lesions (mild and moderate dysplasia) that do not require immediate treatment; it is a challenge to which lesions are at risk for progression. We investigated the association between absence of iodine staining at chromoendoscopy screening and lesion progression using 6 years of follow-up data from a population-based randomized controlled trial in China. We then constructed and validated a model to calculate risk of progression to severe dysplasia, carcinoma in situ, or ESCC. We collected data from 1468 participants (45-69 years old) who were either negative for iodine staining at a baseline chromoendoscopy or found to have mild or moderate dysplasia in histologic analysis of biopsies in the Endoscopic Screening for Esophageal Cancer study in China, from January 2012 through September 2016; 788 of these participants were re-examined by endoscopy after a median interval of 4.2 years (development cohort). We investigated the association between absence of iodine staining and progression of esophageal lesions using Cox prediction models, considering corresponding baseline pathology findings and patient answers to a comprehensive questionnaire. Patients who did not receive a follow-up examination (n= 680) was used as the validation cohort; outcome events in these patients were identified by annual door to door active interviews or linkage with local electronic registry data. The primary outcome was incident esophageal severe dysplasia, carcinoma in situ, or ESCC. In the development cohort, 11 lesions that did not stain with iodine but were classified as not dysplastic in the histology analysis were found to be severe dysplasia, carcinoma in situ, or ESCC at the follow-up evaluation. These lesions accounted for 39.3% of all progressed lesions (n= 28). In the validation cohort, 6 patients with lesions did not stain with iodine but were classified as not dysplastic by histology had a later diagnosis of ESCC, determined from medical records; these patients accounted for 50.0% of all patients with lesion progression (n= 12) until the closing date of this study. We developed a model based on patient age, body mass index, pathology findings, and baseline iodine staining to calculate risk for severe dysplasia, carcinoma in situ, or ESCC. It identified patients for severe dysplasia, carcinoma in situ, or ESCC in the development set with an area under the curve of 0.868 (95% CI, 0.817-0.920) and in the validation set with an area under the curve of 0.850 (95% CI, 0.748-0.952). Almost no cases would be missed if subjects determined to be high or intermediate-high risk subjects by the model were included in surveillance. Absence of iodine staining at baseline chromoendoscopy identifies esophageal lesions at risk of progression with a high level of sensitivity. A model that combines results of iodine chromoendoscopy with other patient features identifies patients at risk of lesion progression with greater accuracy than histologic analysis of baseline biopsies.

Universal endometrial cancer tumor typing: How much has immunohistochemistry, microsatellite instability, and MLH1 methylation improved the diagnosis of Lynch syndrome across the population?

Universal tumor testing for defective DNA mismatch repair (MMR) is recommended for all women diagnosed with endometrial cancer to identify those with underlying Lynch syndrome. However, the effectiveness of these screening methods in identifying individuals with Lynch syndrome across the population has not been well studied. The aim of this study was to evaluate outcomes of MMR immunohistochemistry (IHC), mutL homolog 1 (MLH1) methylation, and microsatellite instability (MSI) analysis among patients with endometrial cancer. A complete systematic search of online databases (PubMed, EMBASE, MEDLINE, and the Cochrane Library) for 1990-2018 was performed. A DerSimonian-Laird random effects model meta-analysis was used to estimate the weighted prevalence of Lynch syndrome diagnoses. The comprehensive search produced 4400 publications. Twenty-nine peer-reviewed studies met the inclusion criteria. Patients with endometrial cancer (n=6649) were identified, and 206 (3%) were confirmed to have Lynch syndrome through germline genetic testing after positive universal tumor molecular screening. Among 5917 patients who underwent tumor IHC, 28% had abnormal staining. Among 3140 patients who underwent MSI analysis, 31% had MSI. Among patients with endometrial cancer, the weighted prevalence of Lynch syndrome germline mutations was 15% (95% confidence interval [CI], 11%-18%) with deficient IHC staining and 19% (95% CI, 13%-26%) with a positive MSI analysis. Among 1159 patients who exhibited a loss of MLH1 staining, 143 (13.7%) were found to be MLH1 methylation-negative among those who underwent methylation testing, and 32 demonstrated a germline MLH1 mutation (2.8% of all absent MLH1 staining cases and 22.4% of all MLH1 methylation-negative cases). Forty-three percent of patients with endometrial cancer who were diagnosed with Lynch syndrome via tumor typing would have been missed by family history-based screening alone. Despite the widespread implementation of universal tumor testing in endometrial cancer, data regarding testing results remain limited. This study provides predictive values that will help practitioners to evaluate abnormal results in the context of Lynch syndrome and aid them in patient counseling.

Mutation pattern in 606 Duchenne muscular dystrophy children with a comparison between familial and non-familial forms: a study in an Indian large single-center cohort

Duchenne muscular dystrophy (DMD) is induced by a wide spectrum of mutations such as exon deletions, duplications and small mutations in the dystrophin gene. This is the first study on the mutational spectrum in a cohort of DMD children from India, with an emphasis to compare the mutations in familial and sporadic forms. Multiplex ligation-dependent probe amplification (MLPA) and next-generation sequencing (NGS) identified 525 and 70 cases of DMD, respectively, while 11 cases showed absent dystrophin staining with no mutations detected. Families with two or more affected males contributed to 12% of the entire cohort. The mutations comprised of exonic deletions in 492/606 (81.2%), duplications in 33/606 (5.4%) and small mutations (point mutations and INDELs) in 70/606 (11.5%) cases. MLPA identified significantly more larger mutations in sporadic (88.2%) than in familial cases (75.3%). The mutations in NGS were: [nonsense = 40 (57.1%); frameshift = 17 (24.3%); splice variant = 12 (17.1%)]. Nonsense mutations were more common in familial than in sporadic cases: 17.8% vs 10.7%. The familial group reported an earlier onset of disease (2.8 ± 1.7years) as compared to sporadic cases (3.8 ± 1.6years). MLPA could identify mutations in a high percentage of our DMD children. The preponderance of small mutations was noted to be distinctly higher in the familial group. Intriguingly, the familial form of DMD formed a small percentage of the entire cohort. The reasons could be increasing awareness among parents and physicians with early identification of DMD cases, genetic counseling and prenatal testing.

Universal endometrial cancer tumor typing: How much has immunohistochemistry, microsatellite instability, and MLH1 methylation improved the diagnosis of Lynch syndrome across the population?

e17119 Background: Universal tumor testing for defective DNA mismatch repair (MMR) is recommended for all women diagnosed with endometrial cancer (EC) to identify those with underlying Lynch syndrome (LS). However, since its implementation in 2013, the effectiveness of this screening method on identifying individuals with LS across the population has not been well studied. The aim of this study was to evaluate outcomes of MMR immunohistochemistry (IHC), MLH1 methylation, and microsatellite instability (MSI) analysis among EC patients. Methods: We conducted a complete systematic search of online databases PubMed, Embase, Medline, and Cochrane Library between 1990-2018. A DerSimonian–Laird random-effects model meta-analysis was utilized to estimate the weighted prevalence of LS diagnoses. Results: The comprehensive search produced 3,427 publications. 29 peer-review studies met the inclusion criteria. 6,649 EC patients were identified, 206 (3%) were confirmed to have LS following positive universal tumor molecular screening.5,917 patients underwent tumor IHC, 28% had abnormal staining. 3,140 patients underwent MSI analysis, 31% had MSI instability. Among EC patients with deficient IHC staining or positive MSI analysis, the weighted prevalence of LS was 15% and 19% respectively. 1159 patients exhibited loss of MLH1 staining, 143 (13.7%) were found to be MLH1 methylation negative, 32 demonstrated a germline MLH1 mutation (2.8% of all MLH1 absent staining; 22.4% of all MLH1 methylation negative). 43% of EC patients diagnosed with LS via tumor typing would have been missed by family history-based screening alone. Conclusions: Despite widespread implementation of universal tumor testing in EC, data regarding results have previously been limited. For the first time, this study provides large-scale predictive values that will help practitioners evaluate abnormal results in the context of LS and aid in patient counseling. [Table: see text]

Evaluation of e-cadherin expression in gastric cancer as a prognostic marker

Introduction: Carcinoma of stomach is a common malignancy and globally stands at fourth position frequency- wise. E-cadherin, is a major cell adhesion molecule, the loss of which gives rise to invasive and metastatic properties to the cancer cells. Aim of the Study: To study the E-cadherin expression in gastric carcinomas by immuno-histochemistry and to correlate its pattern of staining with tumor differentiation, invasion, histological variants, and lymph node status. Materials and Methods: A two year prospective study was done from September 2011 to August 2013 at Kakatiya Medical College and MGM Hospital, Warangal. The resected specimens of stomach and endoscopic biopsy specimens taken from carcinoma stomach in this study period were considered. Routine histopathology and immunohistochemistry for E-cadherin were done on the sections. Results: 60 cases were studied of which 26 were gastric biopsies and 34 were gastrectomy specimens. The patient age ranged from 41-72 years and the male to female ratio was 1.6:1. There were 70% and 30% cases that showed intestinal type and diffuse type morphology respectively. Preserved E-cadherin was mainly seen in intestinal type tumors 14/42 (33.33%) with grade 3 staining, while majority of diffuse adenocarcinomas 8/18 (44.44%) were negative for E-cadherin staining. Higher stage tumors and poorly differentiated adenocarcinomas showed reduced E-cadherin staining. Cases with positive lymph nodes 22/29(75.86%) had diminished / absent E-cadherin staining. Conclusion: Normal gastric mucosa on IHC shows strong membranous E-cadherin positivity. There is a gradual decrease in intensity and percentage, and the pattern changes towards cytoplasmic staining in gastric carcinomas. Keywords: Gastric carcinoma, E –Cadherin expression, Prognostic markers, Immunohistochemistry.

AIP-mutated acromegaly resistant to first-generation somatostatin analogs: long-term control with pasireotide LAR in two patients.

Acromegaly is a rare disease due to chronic excess growth hormone (GH) and IGF-1. Aryl hydrocarbon receptor interacting protein (AIP) mutations are associated with an aggressive, inheritable form of acromegaly that responds poorly to SST2-specific somatostatin analogs (SSA). The role of pasireotide, an SSA with affinity for multiple SSTs, in patients with AIP mutations has not been reported. We studied two AIP mutation positive acromegaly patients with early-onset, invasive macroadenomas and inoperable residues after neurosurgery. Patient 1 came from a FIPA kindred and had uncontrolled GH/IGF-1 throughout 10 years of octreotide/lanreotide treatment. When switched to pasireotide LAR, he rapidly experienced hormonal control which was associated with marked regression of his tumor residue. Pasireotide LAR was stopped after >10 years due to low IGF-1 and he maintained hormonal control without tumor regrowth for >18 months off pasireotide LAR. Patient 2 had a pituitary adenoma diagnosed when aged 17 that was not cured by surgery. Chronic pasireotide LAR therapy produced hormonal control and marked tumor shrinkage but control was lost when switched to octreotide. Tumor immunohistochemistry showed absent AIP and SST2 staining and positive SST5. Her AIP mutation positive sister developed a 2.5 cm follicular thyroid carcinoma aged 21 with tumoral loss of heterozygosity at the AIP locus and absent AIP staining. Patients 1 and 2 required multi-modal therapy to control diabetes. On stopping pasireotide LAR after >10 years of treatment, Patient 1’s glucose metabolism returned to baseline levels. Long-term pasireotide LAR therapy can be beneficial in some AIP mutation positive acromegaly patients that are resistant to first-generation SSA.

Comparison of Papanicolaou Smear Quality with the Anatomical Spatula and the Cytobrush-Spatula: A Single-Blind Clinical Trial.

Background:The Papanicolaou (Pap) smear is a standard test for cervical cancer screening; however, the most important challenge is high false-negative results due to inadequate sampling using the Ayres spatula. The cytobrush has been used in combination with the Ayres spatula (cytobrush–spatula) in an attempt to improve the quality of smears with additional costs. The aim of this study was to compare the Pap smear quality with the anatomical spatula (with extended tip) and the cytobrush–spatula.Materials and Methods:This was a prospective single-bind clinical trial. One hundred and ten sexually active women aged between 22 and 65 years were randomized into groups, each having two smears at the same time: one with a cytobrush–spatula and another with an anatomical spatula. Fifty-five patients were randomized to have the anatomical spatula first to obtain their smears and 55 were randomized to have the cytobrush–spatula first to obtain their smears. Slides were assessed by a pathologist.Results:There was no significant difference in the quality of the smears using the two devices with respect to cellular adequacy (P = 0.3532), absent blood staining (P = 0.7766), presence of endocervical cells (P = 0.3502), and evidence of transformation zone sampling using the Bethesda criteria (0.4028). Kappa analysis shows moderate inter-rater agreement between the two devices by ability to show evidence of transformation zone using British Society for Clinical Cytology and Bethesda criteria.Conclusions:There was no significant difference in the quality of smears obtained using the two different methods. The anatomical spatula can be used as a single device in conventional cytology in place of the cytobrush–spatula with the aim of improving the quality of smears without necessarily increasing the cost.