Immunohistochemical Staining Patterns Research Articles

Prevalence of mesothelin expression in peritoneal disease from colorectal and appendiceal cancers.

Mesothelin is a cell surface glycoprotein overexpressed in 28%-58% of colorectal cancer (CRC). We hypothesized that CRC mesothelin expression contributes to peritoneal spread and that it is selectively overexpressed in those with peritoneal metastasis versus distant metastasis. This case-controlled study involved mesothelin immunohistochemistry staining of tumor specimens from patients with metastatic CRC/appendiceal cancers between 2017 and 2019. Staining reactivity was graded from trace to 4+ (low ≤1+; high >1+). Staining patterns were characterized on global (focal/patchy/diffuse) and cellular (apical/cytoplasmic) levels. Immunostaining of normal mesothelial cells served as internal control. Thirty-one patients were identified: 11 peritoneal (study) and 20 distant metastasis (control). The control group did not include appendiceal cancers. The study group had greater proportion of high staining reactivity (55% vs. 5%; odds ratio [OR] = 20.4, 95% confidence interval [CI] 1.96-211.8). The study group had more diffuse (36% vs. 0%; OR = 22.2, 95% CI 1.1-465.3) and cytoplasmic staining patterns (73% vs. 28%; OR = 6.9, 95% CI 1.3-37.2). Mesothelin expression is higher in CRC/appendiceal cancers with peritoneal metastasis than those with distant metastasis. Immunohistochemistry staining patterns suggestive of propensity towards peritoneal metastasis include diffuse and cytoplasmic staining. Mesothelin may be a potential target for novel treatments of CRC/appendiceal carcinoma with peritoneal involvement.

SIRT4 and SIRT6 Serve as Novel Prognostic Biomarkers With Competitive Functions in Serous Ovarian Cancer.

Sirtuins (SIRTs) are class III histone deacetylases (HDACs) that include seven members and are widely expressed in mammals. Accumulating evidence shows that sirtuins may have contradictory roles in various malignancies. They mainly participate in metabolic homeostasis, DNA damage repair, cell survival, and differentiation, as well as other cancer-related biological processes. To better understand their prognostic role and biological functions, we used comprehensive bioinformatic analyses to demonstrate the expression and mutation of sirtuin family member genes in ovarian cancer (OC), with a detailed focus on prognostic prediction, including the effectiveness of anti-OC drugs. Furthermore, the co-expression genes of SIRT4 and SIRT6 with contradictory survival prediction values in both overall and progression-free survival (PFS) times were further analyzed through Gene Ontology enrichment and Kyoto Encyclopedia annotation. Additionally, we performed and obtained the immunohistochemical staining patterns of these two biomarkers from the serous OC patient database and clinical patient samples to demonstrate their potential applicability in clinical pathology. According to our findings, SIRT4 and SIRT6 are novel prognostic biomarkers that may serve as contradictory competitors for OC cell survival. They are also sensitive biomarkers for the prediction of Avastin’s anticancer effect. While SIRT4 is related to the immune response during oocyte maturation, SIRT6 participates in immune-related disease pathways and mitochondrial metabolism-mediated DNA translation. These findings contribute to the novel hypothesis that SIRT4 and SIRT6 act as contradictory competitors in the regulation of OC behavior. Further studies are required to validate our hypothesis.

GLUT1 Expression in Cutaneous Sebaceous Lesions Determined by Immunohistochemical Staining Patterns.

GLUT1 is a membrane associated carrier protein that functions in the physiologic transport of glucose across cell membranes. Multiple studies have shown an increased GLUT1 expression in various tumor types and a role in cancer prognosis. The aim of this study was to determine whether cutaneous sebaceous lesions have a differential expression of GLUT1 by immunohistochemistry (IHC). GLUT1 IHC was performed on excision specimens of ten cases of sebaceous carcinoma, nine of sebaceoma, ten of sebaceous adenoma, and ten of sebaceous hyperplasia. Intense, diffuse cytoplasmic staining was observed in sebaceous carcinoma. The pattern of GLUT1 staining in sebaceomas and sebaceous adenomas consisted of a gradient of intense cytoplasmic staining in the basaloid cells with a decreased intensity to membranous staining only and absent staining in mature sebaceous cells. In lesions of sebaceous hyperplasia, GLUT1 staining outlined the basal layer of each gland; cytoplasmic staining was minimal to absent. Increased cytoplasmic staining of GLUT1 may correlate with cellular metabolic and proliferative activity. GLUT1 has potential utility in differentiating sebaceous lesions.

OPPORTUNISTIC SURVEILLANCE OF CAPTIVE AND FREE-RANGING BIGHORN SHEEP (OVIS CANADENSIS) IN COLORADO, USA, FOR TRANSMISSIBLE SPONGIFORM ENCEPHALOPATHIES

Bighorn sheep (Ovis canadensis) are predicted to have a degree of susceptibility to the transmissible spongiform encephalopathies (TSE) chronic wasting disease and scrapie. We opportunistically screened 127 captive bighorn sheep and 152 free-ranging bighorn sheep in Colorado, US for the presence of TSE over a period of 35 yr. None of the animals demonstrated clinical signs, gross pathology, histopathology, or immunohistochemical staining patterns suggestive of TSE.

Predictive Patterns of Glutamine Synthetase Immunohistochemical Staining in CTNNB1-mutated Hepatocellular Adenomas.

Some hepatocellular adenoma (HCA) subtypes are characterized by different CTNNB1 mutations, leading to different beta-catenin activation levels, hence variable immunostaining patterns of glutamine synthetase (GS) expression, and different risks of malignant transformation. In a retrospective multicentric study of 63 resected inflammatory (n=33) and noninflammatory (n=30) molecularly confirmed CTNNB1-mutated b-(I)HCA, we investigated the predictive potential of 3 known GS patterns as markers for CTNNB1 exon 3, 7/8 mutations. Pattern 1 (diffuse homogenous) allowed recognition of 17/21 exon 3 non-S45 mutated b-(I)HCA. Pattern 2 (diffuse heterogenous) identified all b-(I)HCA harboring exon 3 S45 mutation (20/20). Pattern 3 (focal patchy) distinguished 12/22 b-(I)HCA with exon 7/8 mutations. In exon 3 S45 and 7/8 mutations, both b-HCA and b-IHCA showed a GS+/CD34- rim with diffuse CD34 positivity in the center of the lesion. Interobserver reproducibility was excellent for exon 3 mutations. Comparative analysis of GS patterns with molecular data showed 83% and 80% sensitivity (b-HCA/b-IHCA) and 100% specificity for exon 3 non-S45. For exon 3 S45, sensitivity was 100% for b-(I)HCA, and specificity 93% and 92% (b-HCA/b-IHCA). For exon 7/8, sensitivity was 55% for both subtypes and specificity 100% and 96% (b-HCA/b-IHCA). Preliminary data from 16 preoperative needle biopsies from the same patients suggest that this panel may also be applicable to small samples. In surgically resected HCA, 2 distinct GS patterns can reliably predict CTNNB1 exon 3 mutations, which are relevant because of the higher risk for malignant transformation. The third pattern, although specific, was less sensitive for the identification of exon 7/8 mutation, but the GS+/CD34- rim is a valuable aid to indicate either an exon 3 S45 or exon 7/8 mutation.

Molecular profiling of the colon cancer in South-Eastern Romania: Results from the MERCUR study.

Colorectal cancer is a heterogeneous disease with multiple epigenetic alterations and different molecular features. The molecular classification is based on 2 major distinct pathways: microsatellite stable pathway and the microsatellite instability pathway. Molecular profiling of colorectal cancer provides important information regarding treatment and prognosis. Aim of the study was to assess the frequency of microsatellite instability in colon cancer and the clinicopathological characteristics of the tumors with high level of microsatellite instability (MSI-H) in our region. The secondary outcome was to assess the frequency of v-raf murine sarcoma viral oncogene homolog B1 (BRAF) mutations in colon cancer.The study included 129 patients with colon cancer fit for surgery. Demographic data, clinical and pathological data, immunohistochemistry staining pattern (4 mismatch repair proteins were investigated), and BRAF gene mutations were assessed. According to microsatellite instability status by polymerase chain reaction, patients were divided into 3 groups: microsatellite stable (MSS) = 108 patients, high level of microsatellite instability (MSI-H) = 15 patients and low level of microsatellite instability (MSI-L) = 6 patients. Different clinicopathological comparisons between MSS and MSI-H patients, and between MSS and MSI-L patients were performed.Microsatellite instability was found in 16.3% patients: 11.6% had MSI-H and 4.7% had MSI-L. Significantly more patients in the MSI-H group than in the MSS group were female (P = .01) and had a family history of colon cancer (P < .001). MSI-H and MSI-L groups were associated with the ascending colon location of the tumors, were mostly type G3, T2, and stage I whereas MSS tumors were mostly G2, pT3, and stage III. Overall, BRAF mutations were identified in 18/129 patients (13.9%). BRAF mutant tumors were predominantly associated with MSI-H and MSI-L tumors. Immunohistochemistry had a sensitivity of 76% and a specificity of 89% in detecting MSI tumors and an accuracy of 87.6%.The frequency of microsatellite instability in our study was 16.3%. MSI-H is a distinct molecular phenotype of colon cancer with particular features: female gender, family history of colorectal cancer, a predilection for the ascending colon, poorly differentiated, predominantly T2, and stage I. The frequency of BRAF mutations was 13.9% and mutations were more often present in the MSI tumors.

Renal angiomyolipoma with epithelial cyst.

ABSTRACTAngiomyolipoma with epithelial cysts (AMLEC) is a recently described entity and is an uncommon subtype of kidney angiomyolipomas. AMLEC is a benign entity but usually masquerades a renal cell carcinoma on imaging examination. AMLEC has a distinct histological and immunohistochemical staining pattern, which helps in the pathological diagnosis. We present a rare case of AMLEC in a 26-year-old female, which was provisionally diagnosed as renal cell carcinoma on radiology. We also summarize the differential diagnosis of this rare variant, its characteristic features, and a review of the literature.

DeepHistoClass: A Novel Strategy for Confident Classification of Immunohistochemistry Images Using Deep Learning

A multitude of efforts worldwide aim to create a single-cell reference map of the human body, for fundamental understanding of human health, molecular medicine, and targeted treatment. Antibody-based proteomics using immunohistochemistry (IHC) has proven to be an excellent technology for integration with large-scale single-cell transcriptomics datasets. The golden standard for evaluation of IHC staining patterns is manual annotation, which is expensive and may lead to subjective errors. Artificial intelligence holds much promise for efficient and accurate pattern recognition, but confidence in prediction needs to be addressed. Here, the aim was to present a reliable and comprehensive framework for automated annotation of IHC images. We developed a multilabel classification of 7848 complex IHC images of human testis corresponding to 2794 unique proteins, generated as part of the Human Protein Atlas (HPA) project. Manual annotation data for eight different cell types was generated as a basis for training and testing a proposed Hybrid Bayesian Neural Network. By combining the deep learning model with a novel uncertainty metric, DeepHistoClass (DHC) Confidence Score, the average diagnostic performance improved from 86.9% to 96.3%. This metric not only reveals which images are reliably classified by the model, but can also be utilized for identification of manual annotation errors. The proposed streamlined workflow can be developed further for other tissue types in health and disease and has important implications for digital pathology initiatives or large-scale protein mapping efforts such as the HPA project.

Diagnostic Utility of p62 Expression in Intranuclear Inclusions in Thyroid Core Needle Biopsy Specimens.

Core needle biopsy (CNB) has been widely used as an alternative method to ultrasound-guided fine-needle aspiration cytology for histological diagnosis of thyroid specimens. However, nuclear artifactual vacuoles (NuVas) produced during tissue processing can be very difficult and sometimes impossible to distinguish from intranuclear inclusions (NuIns). P62 is an autophagy receptor that recognizes, targets, and eliminates toxic cellular materials during autophagy. Herein, we examined the utility of p62 immunohistochemical staining to detect NuIns in thyroid core needle biopsy specimens. Thirty-five thyroid CNB slides from 32 patients and corresponding resection specimens stained with hematoxylin and eosin were reviewed by two pathologists. The immunohistochemical staining pattern of p62 was used to differentiate NuIns from NuVas. The diameter of each nucleus (A) and NuIn (B) was measured, and the number of p62-expressing NuIn-positive (p62In) cells was counted using 1/2 (B/A) and 1/3 (B/A) criteria. The criterion of 1/3 includes NuIns larger than 1/3 and smaller than 1/2 of the nuclear diameter. The criteria of 1/2 includes NuIns larger than 1/2 of the nuclear diameter. By applying the 1/2 criterion, there were no p62In cells in follicular adenoma (FA) samples. However, in papillary thyroid carcinoma (PTC) samples, 22 of 25 specimens exhibited p62In cells. The sensitivity and specificity to distinguish FA from PTC using the 1/2 criterion were 0.88 and 1.00, respectively. By applying the 1/3 criterion, there was one p62In cell hit in FA samples. However, 23 of 25 PTC specimens showed p62In cells. The sensitivity and specificity to distinguish FA from PTC using the 1/3 criterion were 1.00 and 0.90, respectively. P62 is a useful marker for distinguishing FA and PTC based on CNB specimens. We suggest the 1/2 criteria for identifying p62In cells.

Significance of chromogranin A and synaptophysin in medullary thyroid carcinoma.

Medullary thyroid carcinoma (MTC) is a rare thyroid carcinoma of C-cell deviation that produces and secretes calcitonin (CT) and chromogranin A (CgA) into the blood. Thus, CT and CgA are immunohistochemical and serum markers for MTCs. MTC occurs in sporadic and inheritable forms. The hallmark of inheritable cases in multiple endocrine neoplasm 2 (MEN2) is MTC. MEN2 cases represent 30% through germline RET proto-oncogene mutation and occur in younger ages involving bilateral thyroid lobes. Sporadic cases are 70% and occur in older ages. CgA and synaptophysin (SPY) are the two most widely used and reliable immunohistochemical markers for neuroendocrine tumors, including MTCs. This study aimed to detect different immunohistochemical staining patterns for CgA and SPY between non-symptomatic small lesions and invading larger aggressive tumors in both MEA2 cases and sporadic cases. There was different CgA and SPY immunostaining in MEA2 cases where small tumors (≤0.3 cm) were lesser immunostained for CgA and SPY, despite intense staining for CT, compared to the larger (≥0.5 cm) tumors, stronger immunostained for CgA. There was also different CgA and SPY immunohistochemical staining in sporadic cases between small lesions (≤0.5 cm) and larger tumors (≥1.0 cm). One small sporadic tumor (0.5 cm × 0.3 cm) was strongly and weakly, patchy stained for CgA and SPY, respectively, while larger sporadic tumors were diffusely and strongly stained for CgA and SPY. Therefore, stronger CgA and SPY immunostaining for larger tumors in both MEA2 and sporadic cases may be used as independent, aggressive immunohistochemical markers for MTCs.

TSC/MTOR-mutated eosinophilic renal tumors are a distinct entity that is CK7+/CK20-/vimentin-: a validation study

Unclassified renal cell carcinoma (RCC) accounts for ∼10% of renal tumors, and the most common histologic findingin these cases is eosinophilic cytoplasm. We previously demonstrated that a subset of eosinophilic renal tumors with heterogeneous morphology and immunohistochemical (IHC) staining harbored pathogenic mutations in tuberous sclerosis complex (TSC) or mammalian target of rapamycin (MTOR) as the primary defining mutation. We identified an additional 8 cases of eosinophilic tumors with unusual morphology that were originally diagnosed as chromophobe RCC (CHRCC) or CHRCC, eosinophilic variant. As a comparison, we included four classic CHRCC cases and one CHRCC, eosinophilic variant case. Gross examination revealed solid or solid and cystic patterns. The solid areas were composed of eosinophilic tumor cells divided by congested vessels, whereas the cystic areas were lined by cytologically bland eosinophilic cells with septae containing nests, ribbons, and single eosinophilic tumor cells. The tumor cells had abundant granular eosinophilic cytoplasm with round nuclei and inconspicuous nucleoli. IHC analysis demonstrated diffuse staining for CK7 and negative staining for CK20 and vimentin. Next-generation sequencing identified pathogenic variants in three genes: TSC1, TSC2, and MTOR. They also lacked significant copy number variations in contrast to our control cases. We have demonstrated with our expanded study that cases previously diagnosed as CHRCC or CHRCC, eosinophilic variant with discordant histology and IHC staining patterns may represent a separate subtype of RCC characterized by mutations in the TSC/MTOR pathway.

Molecular characterization of chromophobe renal cell carcinoma reveals mTOR pathway alterations in patients with poor outcome

Chromophobe renal cell carcinoma (chRCC) is a histologically and molecularly distinct class of rare renal tumor. TCGA studies revealed low mutational burden, with only TP53 and PTEN recurrently mutated, and discovered alterations in TERT promoter and in the electron transport chain Complex I genes. However, knowledge on drug targetable genes is limited and treatments at metastatic stage do not follow a molecular rationale. In a large series of 92 chRCC enriched with metastatic cases, we performed an in-depth characterization of mTOR pathway alterations through targeted NGS and immunohistochemistry (IHC) of phospho-S6, tuberin, and PTEN. Mutations in mitochondria, telomere maintenance and other renal cancer related genes and p53 IHC, were also assessed. The impact on metastasis development and disease specific survival was determined, using TCGA-KICH series (n = 65) for validation. mTOR pathway mutations (MTOR, TSC1, TSC2) were present in 17% of primary tumors, most of them being classified as pathogenic. Mutations were associated with positive IHC staining of phospho-S6 and PTEN (P = 0.009 and P = 0.001, respectively) and with chRCC eosinophilic variant (P = 0.039), supporting a biological relevance of the pathway. mTOR pathway mutations were associated with worse clinical outcomes. Survival analysis gave a hazard ratio of 5.5 (P = 0.027), and this association was confirmed in TCGA-KICH (HR = 10.3, P = 0.006). TP53 mutations were enriched in metastatic cases (P = 0.018), and mutations in telomere maintenance genes showed a trend in the same direction. p53 IHC staining pattern was associated with the underlying TP53 defect, and negative PTEN IHC staining (82% of cases) suggested PTEN loss as a chRCC hallmark. In conclusion, our study provides with novel genomic knowledge in chRCC and identifies novel markers of poor survival. Furthermore, this is the first study showing that mTOR pathway mutations correlate with poor prognosis, and may help to identify patients with increased sensitivity to mTOR inhibitors.

HER2 Immunohistochemistry in Invasive Micropapillary Breast Carcinoma: Complete Assessment of an Incomplete Pattern.

Invasive micropapillary carcinoma (IMPC) is a rare variant of breast carcinoma, composed of avascular morula-like tumor clusters surrounded by stromal spaces, which can affect the HER2 immunohistochemical (IHC) staining pattern. The 2013 American Society of Clinical Oncology/College of American Pathologists (ASCO/CAP) HER2 testing guideline suggests moderate to intense but incomplete (basolateral) staining be considered equivocal. To perform a detailed assessment of HER2 IHC staining patterns in IMPC. Hematoxylin-eosin and HER2 IHC slides were retrospectively reviewed to assess the morphology and HER2 IHC characteristics of IMPC. The 2018 ASCO/CAP guideline was applied. The cohort consisted of 187 IMPCs from 181 patients with median age of 58 years. Homogeneous (≥90%) micropapillary component was found in 40% (75 of 187) of cases. Receptor profile was as follows: 75% (140 of 187) ER+ HER2-, 19% (37 of 187) ER+ HER2+, 4% (7 of 187) ER- HER2+, and 2% (3 of 187) ER- HER2-. Of 26 cases with HER2 IHC 3+, 65% (17 of 26) showed a basolateral staining pattern with strong intensity. HER2 fluorescence in situ hybridization (FISH) showed amplification in 26% (17 of 66) of HER2 IHC equivocal cases: 76% (13 of 17) showed basolateral staining pattern and 24% (4 of 17) complete staining, with weak to moderate (2), moderate (14), or moderate to strong (1) intensity. The most frequent staining pattern was basolateral, seen in 49% of cases, including 65% HER2 IHC positive and 76% HER2 IHC equivocal/FISH amplified. If a basolateral pattern and weak to moderate staining is observed in IMPC, alternative testing should be performed to confirm the HER2 status.

In situ c-KIT mRNA quantification of canine cutaneous mast cell tumours and its relationship to prognostic factors.

Cutaneous mast cell tumours (MCTs) are the most frequent malignant skin tumours in dogs. Mutations in the c-KIT proto-oncogene are correlated with the pathogenesis and aggressiveness of MCTs. To date, studies have focused on c-KIT mutations and KIT protein localization, with a general lack of mRNA-level analyses. In this study, c-KIT mRNA expression was investigated in canine MCTs by RNA in situ hybridization (RNA-ISH). Furthermore, we evaluated associations between c-KIT mRNA expression and the histological grade, KIT immunohistochemical staining pattern and other clinicopathological parameters. c-KIT mRNA expression was observed in all MCT samples, appearing as clusters of dots in the cytoplasm of neoplastic cells. A significant correlation was detected between c-KIT mRNA expression (quantified according to the H-score and the percentage of positive cells) and the histological grade (determined using two-and three-tier grading systems; P < .05). We also found a significant positive correlation (all P < .05) between c-KIT mRNA expression and the proliferation indices (mitotic index, Ki-67, and Ag67). However, no significant associations with c-KIT expression from RNA-ISH were found with respect to different KIT staining patterns. Overall, these results demonstrate that c-KIT mRNA expression might be an additional tool for measuring the c-KIT status in canine cutaneous MCTs and could serve as a potential prognostic factor. Further studies should evaluate the prognostic significance of c-KIT mRNA expression in a large and uniform cohort of canine MCTs.

Pan-TRK Immunohistochemistry: An Example-Based Practical Approach to Efficiently Identify Patients With NTRK Fusion Cancer.

Food and Drug Administration-approved TRK inhibitors with impressive overall response rates are now available for patients with multiple cancer types that harbor NTRK rearrangements, yet the identification of NTRK fusions remains a difficult challenge. These alterations are highly recurrent in extremely rare malignancies or can be detected in exceedingly small subsets of common tumor types. A 2-step approach has been proposed, involving a screening by immunohistochemistry (IHC) followed by a confirmatory method (fluorescence in situ hybridization, reverse transcriptase-polymerase chain reaction, or next-generation sequencing) in cases expressing the protein. However, there is no interpretation guide for any of the available IHC clones. To provide a pragmatic update on the use of pan-TRK IHC. Selected examples of the different IHC staining patterns across multiple histologies are shown. Primary literature review with PubMed, combined with personal diagnostic and research experience. In-depth knowledge of pan-TRK IHC will help pathologists implement a rational approach to the detection of NTRK fusions in human malignancies.

Urinary Bladder Transitional Cell Carcinoma with Chordoid Features-A Rare Entity

Abstract Casestudy: Invasive urothelial carcinoma is heterogenous entity and show variant morphologic patterns, of which squamous differentiation is the most common. Other variants have been widely recognized and studied. Invasive urothelial carcinoma with chordoid features is a rare entity with only 16 reported cases to date in English literature. It has a significant morphologic overlap with other vesical and non-vesical myxoid neoplasms, including extra-skeletal myxoid chondrosarcoma and chordoma, which necessitate an extensive workup to reach an accurate diagnosis. To the best of our knowledge we report the 17th case of urothelial carcinoma with chordoid features in a 76-year-old male who underwent a cystoprostatectomy and bilateral pelvic lymph node dissection with confirmed diagnosis of high-grade invasive urothelial carcinoma on biopsy. Grossly, bladder showed a 4.2 x 3.4 x 0.6 cm firm white tumor entirely replacing the anterior wall, extending into perivesicular fat, and invading the prostate gland and seminal vesicles. Microscopically, tumor showed urothelial carcinoma in-situ transitioning into high-grade invasive carcinoma with chordoid features. The tumor characterized by round to elongated epithelial cells and eosinophilic cytoplasm arranged in a single file/complex cord-like architecture, in a prominent myxoid stromal background. No conventional high-grade spindle cell morphology was identified. Pelvic lymph nodes were positive for metastatic disease. Tumor cells were positive for GATA3, Pan-cytokeratin, 34BetaE12, and p63, while negative for AMCAR, supporting the urothelial origin of this tumor and confirming the diagnosis. Invasive urothelial carcinoma with chordoid features is a rare entity, generally present at high stage disease, that can be mistaken for other prominent myxoid stromal neoplasms and requires careful assessment. Although, it can demonstrate similar demographic, clinical and immunohistochemical staining pattern as of conventional urothelial carcinoma, special attention should be given to exclude sarcomatoid features which is an aggressive variant of urothelial carcinoma.

Mismatch Repair Gene Expression In Testicular Germ Cell Tumors And Retroperitoneal Lymph Node Metastasis

Abstract Introduction/Objective Testicular germ cell tumors (TCGT) with somatic mismatch repair pathway defects have shown to have worse prognosis and response to therapy. Immunohistochemical staining pattern of MMR panel was classified in a previous study as low (absent or minimal) and high staining (moderate or high staining), with the low staining pattern suggestive of poor prognosis and high risk of recurrence. Herein, we studied the MMR staining pattern in TGCT and lymph node metastasis. Methods 21 TGCT were included in the study: 10 primary testicular tumors and 11 unrelated retroperitoneal lymph node metastasis. All cases were analyzed for MMR protein expression by immunohistochemistry. Clinical, histopathological and follow-up was obtained in all cases. Retained MLH1, MSH2, MSH6, and PMS2 was defined as nuclear staining. Loss of expression was defined as absence of nuclear staining within tumor cells with retained expression in internal control cells, including benign testicular tissue, stromal cells, and infiltrating lymphocytes. Results All primary testicular tumors were mixed GCT: embryonal carcinomas was present in 5, yolk sac tumor in 3 and teratoma in 6 cases. One (10%) of the primary tumors showed focal loss of MLH1 and PMS2 expression in a focus of embryonal carcinoma. All lymph node metastases consisted of teratoma. Seven (64%) metastatic cases showed scattered areas of focal MLH1 loss, 5 (45%) of which showed also focal loss of PMS2. High staining for MSH2 and MSH6 was detected in all 21 cases. Conclusion In our small cohort, focal loss of MLH1 and PMS2 was detected in 10% of primary TGCT, compared to focal loss of MLH1 and PMS2 is 64% and 45%, respectively, of metastatic tumors, suggesting a possible relationship between heterogeneous MLH1 and PMS2 expression and retroperitoneal lymph node metastasis. The current cohort will be expanded to include additional cases.

Comparison of p53 immunohistochemical staining in differentiated vulvar intraepithelial neoplasia (dVIN) with that in inflammatory dermatoses and benign squamous lesions in the vulva.

Differentiated vulvar intraepithelial neoplasia (dVIN), the precursor lesion to human papillomavirus-independent vulvar squamous cell carcinoma (VSCC), can be difficult to distinguish from vulvar inflammatory dermatoses. Our goal was to determine if p53 could be a useful biomarker for dVIN, by characterizing p53 percentage, intensity and patterns of staining in dVIN and its histological mimics. We studied p53 immunohistochemical staining patterns in 16 dVIN cases and 46 vulvar non-neoplastic squamous lesions [12 lichen sclerosus (LS); seven lichen simplex chronicus; three lichen planus (LP); six psoriasis; 13 spongiotic dermatitis (SPO); and five candidiasis]. dVIN cases were adjacent to a p16-negative invasive VSCC in resection specimens. All dVIN cases showed null-type or moderate to strong uniform p53 staining in >70% of basal cells, with moderate to strong continuous parabasal staining extending to two-thirds of the epidermis. This was in contrast to weak or weak to moderate patchy p53 staining in the majority of other lesions. Moderate to strong and increased basal p53 staining (≥70%) was also observed in a subset of LS cases (5/12, 42%), LP cases (1/3, 33%), and SPO cases (36%, 4/11); however, in all categories, this was limited to the basal layer, and any staining in the parabasal layers was patchy. Strong and uniform p53 staining of basal cells, extending into the parabasal layers, and a complete absence of staining (null type) is useful in distinguishing dVIN from other mimics in the vulva. p53 staining of lesser intensity or quantity, particularly basal overexpression only, overlaps with that in vulvar inflammatory lesions.

Femoral Head Chondrocyte Viability at the Cam Deformity in Patients With Femoroacetabular Impingement Syndrome

Background: Patients with hip pathology, such as femoroacetabular impingement (FAI) or hip dysplasia, are known to sustain chondral delamination injuries identifiable during hip arthroscopy, with an incidence of 44% to 75%. There are studies focused on understanding acetabular chondral flap viability, but there is a dearth of research regarding the viability of femoral head cartilage overlying the cam deformity in FAI. Purpose: To describe the viability and immunohistochemistry staining patterns of femoral head cartilage in the setting of FAI. Study Design: Descriptive laboratory study. Methods: Between September 2018 and August 2019, a single surgeon prospectively collected full-thickness femoral cartilage from cam deformities in 14 patients with FAI undergoing osteoplasty. Samples were assessed for viability and underwent immunohistochemistry staining for collagen type I, collagen type II, and aggrecan. Results: The data set included 14 patients. Twelve samples were assessed for viability and 14 for immunohistochemistry straining. The mean patient age was 34.1 years, and the mean body mass index was 24.69. Mean ± SD chondrocyte viability per patient was 52% ± 11%. At the time of cell isolation, 8 of the 12 patients had viability >50%, with a maximum of 68.2%. This viability increased after a primary culture period, varying from 9 to 13 days, with 10 of 12 samples having viability >90%. The viability mean after the culture period was 94.54% ± 4.89%. Harvested cartilage showed expressions of type I cartilage, type II collagen, and aggrecan in a pattern that is predictable for native cartilage. Conclusion: These data reveal that the cartilage in femoral head cartilage overlying cam deformity—much like that from acetabular chondral flaps—not only has baseline viability >50% (51.99% ± 10.83%) but the ability to increase in viability >90% after a culture period. There may be a role for use of femoral head cartilage as autograft to repair full-thickness cartilage defects of the acetabulum and femoral head, either at the time of osteochondroplasty or after a period of cell culture to improve cell viability. Clinical Relevance: A dearth of information is available regarding the viability of femoral head cartilage. This study provides insight into the cartilage viability and response to culture.

Analysis of Tumor Depth Invasion With Anti-Smoothelin Antibody in Equivocal Transurethral Resection of Urinary Bladder Tumor Surgical Specimens.

To examine the expression and value of the smoothelin marker in control cases, to standardize the working method, and to analyze its application in pathologic staging process of problematic transurethral resection of bladder tumor (TURBT) cases. Immunohistochemical (IHC) staining was performed on tumor-free bladder wall sections, tumor-free large bowel sections, TURBTs with unequivocal tumor stage, and TURBTs with equivocal stage. The IHC staining of muscularis mucosa (MM), muscularis propria (MP), and blood vessels was evaluated semiquantitatively. Smoothelin IHC staining pattern ranged from negative (30% to 67% cases) to 2+ (0% to 15% cases) in MM and from 1+ (10% to 50% cases) to 3+ (9% to 48% cases) in MP. When compared on the same slide, the smoothelin expression of MP showed a stronger staining intensity than the one of the MM in all the analyzed cases. Blood vessel muscle cells stained in a constant intensity as the MM (r = 0.9808; r = 0.9604). Smoothelin determined restaging of 33% of the problematic TURBT cases. Smoothelin is an IHC marker that shows differential staining between coexistent MM and MP; however, variations in staining intensity and pattern may occur, aspects that can be influenced by different technique variables. We recommend using this marker as a diagnostic tool in problematic TURBT cases only when there is sufficient experience in control cases with this antibody.