Paraffin-embedded Tissue Blocks Research Articles

Potential role of BRCA1 protein expression as a prognostic tissue biomarker in breast carcinoma: an immunohistochemical and clinicopathologic study from South India

Introduction. BRCA1 dysfunction is a hallmark of both hereditary and sporadic breast cancer. BRCA1 protein expression can be lost by germline mutation, somatic mutation or promoter hypermethylation. This study aimed to explore BRCA1 dysfunction in breast cancer patients by immunohistochemistry and to study its association with prognostic factors. Material and methods. BRCA1 protein expression was assessed by immunohistochemistry on formalin fixed paraffin embedded tissue blocks of 110 invasive breast carcinoma patients. Furthermore, the clinical findings and tumor features associated with BRCA1 dysfunction were characterized. Results. Reduced BRCA1 immunoreactivity was observed in 19% of breast cancer cases. Although these patients presented with aggressive tumor characteristics, statistical significance was observed only with presence of lymphovascular emboli (p < 0.05). These results suggest that loss of BRCA1 protein expression is associated with an aggressive phenotype of breast carcinoma. Conclusions. Immunohistochemistry for BRCA1 protein expression in tumor tissues may provide a less expensive screening tool to identify BRCA1 dysfunction due to genetic or epigenetic alterations.

Iso-chromosome 12p Analysis by Fluorescent In-Situ Hybridization: An Academic Institutional Experience

Abstract Introduction/Objective Chromosome 12 abnormalities like iso-chromosome 12p (i12p) and amplification of 12p are seen in majority (89%) of the primary and metastatic testicular germ cell tumors (TGCTs). i12p can be detected by karyotyping, fluorescent in-situ hybridization (FISH) or reverse transcriptase polymerase chain reaction. The aim of this study was to review i12p FISH data at our institution and assess the clinical utility. Methods/Case Report Laboratory information system was queried over a period of 15 years to search for cases where i12p FISH test was requested. FISH test was performed using TelVysion 12p telomeric probe and CEP 12 centromere probe on paraffin-embedded tissue or cell blocks. A ratio of 12ptel/CEP12 signal of 1.4 or greater was considered as positive. Patient demographics, clinical presentation, pathologic findings, and follow-up data were documented and correlated. Results (if a Case Study enter NA) Total 58 cases were identified with an age range of 14 to 76 years. Majority were male (M=52, F=6). Of these cases, 15 were testicular and 43 extra-testicular cases that included resection (n=35), biopsy (n=20) and cell-blocks (n=3). i12p was detected in 8 out of 15 testicular cases while i12p was detected in 16 out of the 43 extra-testicular cases. The extra- testicular cases included 17 retroperitoneal lesions, 8 lesions from the mediastinum, 6 lymph nodes from other sites and 12 miscellaneous lesions. Using pathology diagnosis with immunohistochemistry as gold standard, overall sensitivity was 60% and specificity was 86%. There were 3 false positive cases [Benign testicular parenchyma (n=1), suspicious for germ cell neoplasia in-situ (n=1) and undifferentiated epithelioid neoplasm (n=1)]. Conclusion Our results show that although the sensitivity was limited, FISH test for i12p demonstrated high specificity(86%) for diagnosis of primary or metastatic TGCTs. As an adjunct test, i12p FISH can help identify and further characterize a significant number of GCTs with unusual morphology or clinical presentation.

Scientific and Regulatory Policy Committee Points to Consider: Biological Sample Retention From Nonclinical Toxicity Studies.

Samples of biologic specimens and their derivatives (eg, wet tissues, paraffin-embedded tissue blocks, histology slides, frozen tissues, whole blood, serum/plasma, and urine) are routinely collected during the course of nonclinical toxicity studies. Good Laboratory Practice regulationsand/or guidance specify minimum requirements for specimen retention duration, with the caveat that retention of biologic specimens need not extend beyond the duration of sample stability. However, limited availability of published data regarding stability for various purposes following storage of each specimen type has resulted in confusion, uncertainty, and inconsistency as to the appropriate duration for storage of these specimens. To address these issues, a working group of the Society of Toxicologic Pathology Scientific and Regulatory Policy Committee was formed to review published information, regulations, and guidance pertinent to this topic and to summarize the current practices and rationales for retention duration through a survey-based approach. Information regarding experiences reaccessing biologic specimens and performing sample stability investigations was also collected. Based on this combined information, the working group developed several points to consider that may be referenced when developing or revising sample retention practices. [Box: see text].

Evaluation of Angiogenesis by Using CD105 and CD34 in Sudanese Breast Cancer Patients.

<b>Background and Objective:</b> Angiogenesis is a mechanism by which new blood vessels are developed in healing and tumour tissues, where it is necessary for regeneration growth, tumour cells survival and metastasis. This study aimed to assess the angiogenesis mechanism among Sudanese females with breast cancer using anti-CD34 and anti-CD105 markers. <b>Materials and Methods:</b> Three hundred female representative Formalin-Fixed Paraffin-Embedded (FFPE) breast tissue blocks were included in this study. Of the 300 representative tissue blocks, 200 were breast cancer patient's tissues (confirmed cases) and 100 were normal breast tissues (controls). Their ages mean±SD, 47.3±12.9 years. <b>Results:</b> The results showed the MVD of CD34 significantly increased in malignant lesions as compared to normal breast tissues. The mean of MVD CD34 and MVD CD105 showed statistical differences among different histologic types of breast cancer. Also, a strong positive correlation was detected between the manual and automated MVD counting methods. Also, the current study revealed no significant differences were observed in mean MVD counting for both markers and menopausal status or the age groups of the study population. <b>Conclusion:</b> The MVD is a good tool for assessing prognostic markers. The CD105 marker has a high specificity to the new evolving tumour vessels and is a useful predictor for angiogenesis and breast cancer metastasis.

Role of special stains in identification of fungi in eumycetoma among Sudanese patients in Soba University Hospital

Background: Mycetoma is a unique neglected tropical disease caused by a substantial number of microorganisms of fungal or bacterial origins. Identification of the causative organism and the disease extension are the first steps in the management of the affected patients and predicting disease treatment outcome and prognosis. Objectives: To determine the role of special stain in identification of fungi in eumycetoma among Sudanese patients at Soba University Hospital. Method: This descriptive cross- sectional study was conducted of Fifty-nine archival formalin fixed paraffin- embedded tissue blocks, four identical sections from each archival paraffin-embedded tissue block were obtained and stained with routine HE also mycetoma is most common in young adults (16–40 years old) and is uncommon in children. No significant relationship was demonstrated between diagnosis and variables (gender, age). Type I and II was the commonest type of the host tissue reaction encountered in this study. Conclusion: Results showed that GMS and PAS had the ability to stain both fungal hyphae and yeast in compare to Gridley’s stain which had some limitations to stain fungal structures.

A Comparative Immunohistochemical Study of Expression of Syndecan-1 (CD138) and Podoplanin in Keratocystic Odontogenic Tumor, Orthokeratinized Odontogenic Cyst and Dentigerous Cyst

Background: Odontogenic lesions can bring about diagnostic challenges due to overlapping histology among pathologists. Thus, there are few tumor markers that provide accurate diagnosis. Syndecan-1 (CD-138) and podoplanin (PDPN) are proteoglycans that have been described as substantial diagnostic and prognostic markers in various odontogenic lesions. Aim: This study aims to evaluate and compare the immunohistochemical expression of syndecan-1 (CD-138) and podoplanin in keratocystic odontogenic tumor, orthokeratinized odontogenic cyst (OOC), and dentigerous cyst (DC). Materials and Methods: The formalin-fixed paraffin-embedded tissue blocks of KCOT, OOC, and DCs were retrieved form the archives of department. Three sections each of 3 μm thickness were made using a rotary microtome and they were stained with syndecan-1 (CD-138) and podoplanin (PDPN) using immunohistochemical methods and standard hematoxylin and eosin stain. Results: Immunohistochemical expression of syndecan-1in KCOT was found to be weakly to moderately positive in 6 cases with 2 cases exhibiting positive expression. In OOC, 3 cases displayed negative expression-1 whereas 7 cases were weakly to moderately positive. Immunohistochemical expression of podoplanin in KCOT was observed to be weakly to moderately positive in 4 cases with 5 cases exhibiting strongly positive expression. In OOC, 3 cases displayed negative immunohistochemical expression for podoplanin and 4 cases were strongly positive. Immunoreactivity for podoplanin in DC was negative in 3 cases whereas 5 cases were weakly to moderately positive. Conclusion: The absence of significant correlation between expression of syndecan-1 and podoplanin reinforces the exact role of these proteins in the differentiation of odontogenic lesions which need to be elucidated further.

Utilization of cell proliferation markers to diagnose cystic jaw pathologies

Markers of cell proliferation are widely used as diagnostic and prognostic tools. Coincident estimation of these markers increases the precise evaluation of the proliferative status of different tissues and can also be helpful in determining progression, aggressiveness and prognosis of the lesions. The current study investigated the expression of PCNA and MCM3 cell proliferation markers in 40 formalin fixed paraffin embedded tissue blocks of odontogenic keratocyst (OKC) and unicystic ameloblastoma (UA) cases using immunohistochemistry method. Markers` expression based on the intensity, percentage of positively stained cells and localization of reaction through the cyst lining epithelium was separately analyzed for each marker using Chi square test, the results of which were significant for the two markers (P < 0.05). Both markers revealed statistically significant differences between OKC and UA cases regarding markers expression intensity, positivity score and localization of reaction through the epithelium. Mural UA histologic variant was significantly different than luminal and intraluminal variants. The correlation coefficient between the two markers was found to be 0.86.

A Cross-Sectional Study for Evaluation of KRAS and BRAF Mutations by Reverse Dot Blot, PCR-RFLP, and Allele-Specific PCR Methods Among Patients with Colorectal Cancer

KRAS and BRAF genes are the biomarkers in Colorectal Cancer (CRC) which play prognostic and predictive roles in CRC treatment. Nowadays, the selection of rapid and available methods for studying KRAS and BRAF mutations in anti-EGFR therapy of patients suffering from CRC plays a significant role. In this study, the mutations of these two oncogenes were evaluated by different methods. This study was performed on 50 Formalin-Fixed Paraffin-Embedded (FFPE) tissue blocks of patients diagnosed with colorectal cancer. After DNA extraction, KRAS and BRAF gene mutations were evaluated using reverse dot blot, and results were compared with PCR-RFLP and allele-specific PCR for KRAS and BRAF mutations, respectively. KRAS gene mutations were detected in 42% of patients, of which 30% were in codon 12 region, and 12% in codon 13. The most frequent mutations of KRAS were related to G12D and 10% of patients had BRAF mutated genes. The type of KRAS gene mutations could be evaluated by reverse dot blot method. In general, the results of PCR-RFLP and allele-specific PCR were similar to the findings by reverse dot blot method. These findings suggest that PCR-RFLP and allele-specific PCR methods are suitable for screening the presence of the mutations in KRAS and BRAF oncogenes. In fact, another method with more sensitivity is needed for a more accurate assessment to determine the type of mutations. Due to higher speed of detection, reduced Turnaround Time (TAT), and possible role of some KRAS point mutations in overall survival, reverse dot blot analysis seems to be an optimal method.

A comparative immunohistochemical analysis of cortactin in orthokeratinized odontogenic cyst (OOC), sporadic odontogenic keratocyst (OKC), and syndromic OKC

ObjectivesTo evaluate and compare the immunohistochemical expression of cortactin in the epithelial lining of orthokeratinized odontogenic cyst (OOC), sporadic odontogenic keratocyst (OKC), and syndromic OKC. MethodsFormalin-fixed paraffin-embedded tissue blocks of histopathologically diagnosed cases of OOC, OKC, syndromic OKC, normal buccal mucosa (NBM), and oral squamous cell carcinoma (OSCC) were examined for immunohistochemical expression of cortactin. Clear brown cytoplasmic and membranous staining was considered positive. ResultsA statistically significant difference was observed between OOC and syndromic OKC (p < 0.001), as well as between sporadic OKC and syndromic OKC (p < 0.001). Although not statistically significant, the expression of cortactin was slightly higher in the basal layer of NBM (mean = 0.47), OOC (mean = 0.27), sporadic OKC (mean = 0.47) syndromic OKC (mean = 1.53), and OSCC (mean = 0.67) than in the parabasal layers of NBM (mean = 0.27), OOC (mean = 0.20), sporadic OKC (mean = 0.47), syndromic OKC (mean = 1.27), and OSCC (mean = 0.60). ConclusionThe expression of cortactin in the basal layer may suggest the formation of invadopodia in the basal layer where the invasion mechanism occurs. This finding is further supported by the higher localization of cortactin in areas of epithelial budding and daughter cysts in syndromic OKC, thereby reaffirming its possible association with recurrence.

Study of expression of endoglin (CD105) in oral squamous cell carcinoma.

Context:Oral cancer is the 8th most common cancer in the world. An important feature of carcinogenesis is angiogenesis. Endoglin is a powerful marker of neovascularization in solid malignancies. This study was done to ascertain its role as an indicator of metastasis and prognosis.Aim:This study aimed to evaluate and compare the expression of endoglin (CD105) in metastatic primary tumor, lymph node of the metastasized tumor, nonmetastatic primary tumor and in normal buccal mucosa immunohistochemically.Settings and Design:The total sample size comprised 45 formalin-fixed paraffin-embedded tissue blocks, n = 10 metastasized primary tumor, n = 10 lymph nodes of metastasized primary tumor, n = 20 nonmetastasized oral squamous cell carcinoma and n = 5 normal buccal mucosa were studied.Subjects and Methods:Immunohistochemistry for endoglin was performed and microvessel density (MVD) was determined by hot spot method. Microvessel density was compared between the groups.Statistical Analysis:Statistical analysis was used using one-way ANOVA. P < 0.05 was statistically significant.Results:Endoglin expression in metastatic cases (0.68 + 0.10) was higher than nonmetastatic cases (0.45 + 0.20) and the difference was statistically significant (P = 0.002).Conclusion:This study shows that presence of endoglin determines the metastatic potential of the tumor and its prognosis, thus, could be considered as a potential target of therapy.

MP66-14 DEVELOPMENT &amp; VALIDATION OF A METHOD FOR DETECTION OF CIRCULATING TUMOUR DNA IN MUSCLE-INVASIVE BLADDER CANCER

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP66)1 Sep 2021MP66-14 DEVELOPMENT & VALIDATION OF A METHOD FOR DETECTION OF CIRCULATING TUMOUR DNA IN MUSCLE-INVASIVE BLADDER CANCER Ruchira Nandurkar, Pavel Sluka, Hady Wardan, Ian Davis, and Shomik Sengupta Ruchira NandurkarRuchira Nandurkar More articles by this author , Pavel SlukaPavel Sluka More articles by this author , Hady WardanHady Wardan More articles by this author , Ian DavisIan Davis More articles by this author , and Shomik SenguptaShomik Sengupta More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002106.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Survival from muscle-invasive Bladder Cancer (MIBC) remains variable despite optimal therapy. Current clinical and pathological variables are limited in their prognostic and predictive utility. Identifying the presence and amount of circulating tumour DNA (ctDNA) in the bloodstream is of great interest as an additional tool. Currently, this relies on next-generation sequencing of tumour samples, which is expensive and can take a long time for results, thus precluding responsive clinical decision-making. This study aimed to develop a quick, sensitive, inexpensive and reliable method for detection of ctDNA in MIBC using droplet digital PCR (ddPCR). METHODS: We selected a combination of 10 mutations found in MIBC that were not only common, but when sequentially added to a gene panel, each would increase the panel’s capacity to detect a wider cohort of MIBC patients, thereby maximising the detection rate whilst maintaining assay feasibility. Mutations were selected based upon data published by The Cancer Genome Atlas (TCGA). Our 10-gene panel was tested against archival MIBC specimens. DNA extracted from archival paraffin-embedded tissue blocks was run against the gene panel using ddPCR to assess for presence and fractional abundance of the mutation in each sample. RESULTS: To date, 66 MIBC samples have been processed, of which 65 possessed at least one of the mutations within our gene panel, conferring a 98.5% detection rate, with a median of five panel mutations detected per sample (range 0-9). Our use of ddPCR may be accountable for our higher detection rate compared to the 40% predicted from the TCGA, which used Next Generation Sequencing technology. Further analyses are being undertaken to compare samples before and after neo-adjuvant chemotherapy. CONCLUSIONS: This technique utilising a 10 gene panel appears to be sensitive enough to detect mutations in the majority of patients with MIBC. Prospective validation is underway to determine the potential clinical utility of this technique. Source of Funding: Robert Bulley Research and Innovation Grant, Eastern Health Foundation © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1134-e1135 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ruchira Nandurkar More articles by this author Pavel Sluka More articles by this author Hady Wardan More articles by this author Ian Davis More articles by this author Shomik Sengupta More articles by this author Expand All Advertisement Loading ...

FOXA1 is associated with high tumor grade, myometrial invasion and lymph node invasion in endometrial endometrioid carcinoma.

FOXA1 expression has been demonstrated in several hormone-dependent cancers. However, data are limited concerning the role of FOXA1 in endometrial cancers. The present study aimed to investigate FOXA1 expression via the microarray technique in benign hyperplasia, endometrial intraepithelial neoplasia, and endometrial endometrioid carcinoma. We also aimed to determine whether there were any associations between FOXA1 expression, tumor grade, myometrial invasion and lymphatic invasion. Paraffin-embedded sections prepared from samples obtained from 114 patients who underwent surgical hysterectomy or curettage were analyzed. Data were retrieved from digitally-stored medical records. Tissue microarrays were prepared from formalin-fixed, paraffin-embedded tissue blocks. Full tumor sections were used for immunohistochemical analysis performed. Carcinomas with nuclear grade 3 had higher FOXA1 values than others, while grade 2 carcinomas also had higher FOXA1 values relative to grade 1 (p < 0.001). FOXA1 values of FIGO stage III carcinomas were significantly higher than others and stage II values were also significantly higher than stage I FOXA1 values (p < 0.001). Patients with myometrial and lymph node invasion had significantly higher FOXA1 values than others (p < 0.001 and p = 0.047, respectively). FOXA1 had 91.30% sensitivity, 63.60% specificity and 77.78% accuracy for predicting the presence of myometrial invasion with a cut-off value of 9. FOXA1 expression is higher in endometrial endometrioid carcinoma compared to benign endometrial hyperplasia or intraepithelial neoplasia. In patients with endometrial endometrioid carcinoma, high FOXA1 expression is associated with high tumor grade, myometrial and lymph node invasion. However, FOXA1 expression is not associated with lymphovascular or cervical invasion.

Diagnosis of Primary Cutaneous Amyloidosis by Rapid 4,6-Diamidino-2-Phenylindole Staining

Background: Quick and accurate diagnosis of primary cutaneous amyloidosis (PCA) may be difficult because its symptoms are often subtle and nonspecific. Objective: We sought to review the literature on the roles of various staining methods in the diagnosis of amyloidosis and demonstrate added benefits of using rapid 4,6-diamidino-2-phenylindole (DAPI) staining in the diagnosis of PCA. Methods: Three groups of cases, namely, PCA, neurodermatitis, and prurigo nodularis, were retrieved from a computerized pathology database for study, and their paraffin-embedded tissue blocks were cut following standard procedures. The tissue sections were stained with three stains: hematoxylin-eosin (HE), Congo red, and DAPI stains, and examined under the microscope to compare the staining patterns of these three methods. We also performed amyloid keratin and apolipoprotein E (APOE) staining on the sections of PCA in order to further support our conclusion. The PCA sections were read by junior and senior dermatopathologists for comparison. Results: The sensitivity of DAPI staining for PCA was significantly higher than that of Congo red staining and HE staining (p < 0.001). This statement holds true whether the experiment was grouped in one sample or was divided into groups of junior and senior dermatopathologists (p < 0.001). The DAPI-positive staining areas, except for the nuclei, were consistent with the amyloid deposition areas. In this study, DAPI staining had a sensitivity of 98.6% and a specificity of 100%. Conclusion: DAPI staining could serve as a useful technique to establish the diagnosis of PCA, and its high efficacy in diagnosing PCA makes it less dependent on the experience levels of the evaluators. Additionally, the binding of DAPI to the A-T-rich sequence of double-stranded DNA suggests that amyloid may contain DNA or a similarly structured nucleic acid.

CCNE1 amplification among metastatic sites in patients with gynecologic high-grade serous carcinoma

ObjectiveWe sought to characterize the variability of CCNE1 amplification among metastatic sites of CCNE1 amplified high grade serous carcinoma (HGSC) cases to investigate the feasibility of targeting this alteration for therapeutic purposes. MethodsPatients with CCNE1 amplified HGSC who underwent surgical cytoreduction with metastatic sites were identified from institutional molecular profiling reports and a population of HGSC cases screened using digital droplet PCR (ddPCR). Cases with normal CCNE1 copy number were included as controls. Slides from metastatic sites were cut from formalin-fixed paraffin-embedded tissue blocks, dissected for tumor of > 50% purity, and underwent DNA extraction. CCNE1 copy number was determined by ddPCR. Tumor purity was confirmed with mutant TP53 allele fraction from targeted massively parallel sequencing. ResultsFour of 15 patients from an institutional database screened by ddPCR were found to have CCNE1 amplification. Three additional patients were identified from a query of institutional commercial clinical reports. Among these 7 CCNE1 amplified cases (2 uterine, 5 ovarian), 5 showed preservation of CCNE1 amplification (copy number > 5) among all metastatic sites. The remaining 2 cases had multiple metastatic sites without preserved CCNE1 amplification. Non-amplified cases had predominantly normal CCNE1 copy number across metastatic sites. ConclusionsCCNE1 amplification is an early genomic event in HGSC and is preserved in most metastatic sites suggesting a uniform response to pathway targeting therapies.

MRNA EXPRESSION OF PD-1, PD-L1, AND IMMUNOTHERAPY IN BLADDER CANCER

Objective: This study aims to investigate mRNA expression of PD-1 and PD-L1 in patients with bladder cancer. Material &amp; Methods: In this study, we examined 30 samples from paraffin embedded tissue blocks, samples were divided into two groups, 15 were NIMBC, and 15 were MIBC according to their histopathological result. mRNA expression of PD-1 and PD-L1 were conducted using Real Time-Polymerase Chain Reaction (qRT-PCR) and statistical significance was set at a p-value &lt; 0.05. Results: Statistical analysis using the Mann-Whitney test found a significant difference in mRNA expression of PD-1 and PD-L1 in NMIBC compared to MIBC groups. Conclusion: mRNA expression of PD-1 and PD-L1 were higher in MIBC compared to NMIBC. PD-1 and PD-L1 as immune checkpoints are potential immunotherapy for patients with advance stage bladder cancer. Immunotherapy could be a substitute or combined with other treatments such as chemotherapy or radiotherapy.

BRAF V600E and Mismatch Repair Proteins Expression in Sporadic Young-onset Colorectal Cancer in Kelantan, Malaysia.

ObjectivesWe sought to determine the immunohistochemistry expression of mismatch repair (MMR) and BRAF V600E proteins in sporadic young-onset colorectal cancer (CRC) and their association with clinicopathological features in the Kelantan population.MethodsThis was a cross-sectional study of sporadic young-onset CRC over 11 years from 1 January 2006 to 31 December 2017 in Kelantan. Formalin-fixed paraffin-embedded tissue blocks were immunohistochemically stained with antibodies for MMR (MLH1, MSH2, MSH6, and PMS2) and BRAF V600E. These expressions were correlated with clinicopathological parameters.ResultsOur patient sample included 31 patients with a mean age of 31.5 years. More than half (61.3%) of the patients were women. The majority presented with abdominal pain (41.9%), and 71.0% had a tumor located on the right side of the colon, with 83.9% being moderately differentiated adenocarcinoma. The majority of patients presented at stage IV (54.8%). The most frequent pattern was all MMR protein expressions, which constituted patients in the microsatellite stable group (64.5%). Nine (29.0%) were microsatellite instability (MSI-high), and two (6.5%) were MSI-low. Positive BRAF V600E expression was observed in 83.9% of patients. Only histopathological subtypes revealed a significant association with BRAF V600E positive expression (p = 0.015).ConclusionsThe majority of sporadic young-onset CRC presented with abdominal pain and advanced cancer stage. Most were microsatellite stable, and most cases showed positive expressions in all MMR markers and BRAF V600E by immunohistochemistry method. This finding will pave the way for further research on this disease.

Somatic mutations in benign breast disease tissues and association with breast cancer risk

BackgroundBenign breast disease (BBD) is a risk factor for breast cancer (BC); however, little is known about the genetic alterations present at the time of BBD diagnosis and how these relate to risk of incident BC.MethodsA subset of a long-term BBD cohort was selected to examine DNA variation across three BBD groups (42 future estrogen receptor-positive (ER+) BC, 36 future estrogen receptor-negative (ER−) BC, and 42 controls cancer-free for at least 16 years post-BBD). DNA extracted from archival formalin fixed, paraffin-embedded (FFPE) tissue blocks was analyzed for presence of DNA alterations using a targeted panel of 93 BC-associated genes. To address artifacts frequently observed in FFPE tissues (e.g., C>T changes), we applied three filtering strategies based on alternative allele frequencies and nucleotide substitution context. Gene-level associations were performed using two types of burden tests and adjusted for clinical and technical covariates.ResultsAfter filtering, the variant frequency of SNPs in our sample was highly consistent with population allele frequencies reported in 1 KG/ExAC (0.986, p < 1e−16). The top ten genes found to be nominally associated with later cancer status by four of 12 association methods(p < 0.05) were MED12, MSH2, BRIP1, PMS1, GATA3, MUC16, FAM175A, EXT2, MLH1 and TGFB1, although these were not statistically significant in permutation testing. However, all 10 gene-level associations had OR < 1 with lower mutation burden in controls compared to cases, which was marginally statistically significant in permutation testing (p = 0.04). Comparing between the three case groups, BBD ER+ cases were closer to controls in mutation profile, while BBD ER− cases were distinct. Notably, the variant burden was significantly higher in controls than in either ER+ or ER− cases. CD45 expression was associated with mutational burden (p < 0.001).ConclusionsSomatic mutations were more frequent in benign breast tissue from women who did not develop cancer, opening questions of clonal diversity or immune-mediated restraint on future cancer development. CD45 expression was positively associated with mutational burden, most strongly in controls. Further studies in both normal and premalignant tissues are needed to better understand the role of somatic gene mutations and their contribution to future cancer development.

Expression of Progesterone Receptor and Its Association with Clinicopathological Characteristics in Meningiomas: A Cross-Sectional Study

BackgroundMeningiomas that are progesterone receptor positive have a low recurrence rate and good prognosis compared to those that are progesterone receptor negative. This study aimed to determine the prevalence of expression of progesterone in meningiomas and its association with clinicopathological characteristics.Materials and methodsThis was a cross-sectional laboratory-based study that was conducted at Muhimbili National Hospital. The study included 112 formalin-fixed paraffin-embedded tissue blocks of patients who were confirmed to have meningiomas on histological basis from January 2010 to December 2014. Immunohistochemical expression of progesterone receptor was tested using a primary monoclonal progesterone receptor antibody ready to use (IR 068 Dako). The χ2 test was used to determine the association between clinicopathological characteristics and progesterone receptor expression. A 2-tailed P < 0.05 was considered significant.ResultsThe mean age of the patients was 45.5 ± 3.601 years, and majority (66.1%, n = 74) were in the age group between 31 and 60 years. Also, majority of the patients (60%, n = 67) in this study were females. Over one-third of the cases (34.8%, n = 39) comprised of meningotheliomatous subtype, and majority of the cases (89.3%, n = 100) were of grade I. The prevalence of progesterone expression was 54.5% (n = 61), and only age was associated with progesterone receptor expression (P = 0.043).ConclusionThe finding of high expression of the progesterone receptor for grade I cases in this study indicates that progesterone receptor expression in meningiomas is of prognostic value and may be considered when evaluating patients for management. Lack of expression of progesterone receptor in all the malignant cases is intriguing and needs further studies that can investigate its prognostic role.

Significance of Vascular Endothelial Growth Factor Expression in the Bladder Urothelial Carcinoma and Its Association with Tumor Grade and Invasiveness

Background & Objective: Vascular Endothelial Growth Factor (VEGF) is one of the newer molecular markers that acts as a central mediator of tumor angiogenesis and is essential for tumor growth, progression, and metastasis. So anti-angiogenic drugs can be used as anticancer therapy. Treatments with anti-VEGF (Bevacizumab) therapy have been proved to improve relapse-free survival in many tumors. Urinary bladder tumor has become emerging cancer globally among elderly individuals. So, the identification and development of novel biomarkers for effective treatment of urinary bladder carcinoma is essential. The present study aimed to investigate the immunohistochemical expression of VEGF in urothelial carcinoma of urinary bladder and to assess its association with tumor grade and muscle invasiveness.Methods: This cross-sectional study was conducted in the Department of Pathology, Chittagong Medical College, Chattogram from September 2018 to August 2020. Fifty-six formalin-fixed paraffin-embedded tissue blocks of urinary bladder carcinoma were prepared for both histopathological and immunohistochemical examination. Each slide was evaluated by at least two pathologists.Results: Weak to strong positive expression of VEGF were observed in 52 cases (92.86%). The proportion of tumors positive for VEGF expression was higher among patients with high grade and non-muscle invasive bladder carcinoma. Conclusion: We found that VEGF expression has a significant association with tumor grade and an inverse association with muscle invasion. These findings may be useful for selecting the subset of patients likely to respond to anti-VEGF targeted therapy.

Effects of Slide Storage on Detection of Molecular Markers by IHC and FISH in Endometrial Cancer Tissues From a Clinical Trial: An NRG Oncology/GOG Pilot Study.

We performed a pilot study in anticipation of using long-aged precut formalin-fixed paraffin-embedded tissue sections stored in real-world conditions for translational biomarker studies of topoisomerase 2A (TOP2A), Ki67, and human epidermal growth factor receptor 2 (HER2) in endometrial cancer. Formalin-fixed paraffin-embedded tissue blocks or unstained slides or both from GOG-0177 were collected centrally (1999-2000) and stored at room temperature. During 2004 to 2011 specimens were stored at 4°C. Matched pairs of stored slides and freshly cut slides from stored blocks were analyzed for TOP2A (KiS1), Ki67 (MIB1), and HER2 (HercepTest) proteins. To assess DNA stability (HER2 PathVision), fluorescence in situ hybridization (FISH) was repeated on stored slides from 21 cases previously shown to be HER2 amplified. Immunohistochemistry (IHC) staining intensity and extent, mean FISH copies/cell, and copy number ratios were compared using the κ statistic for concordance or signed rank test for differences in old cut versus new cut slides. IHC results reflected some protein degradation in stored slides. The proportion of cells with TOP2A staining was lower on average by 12% in older sections (P=0.03). The proportion of Ki67-positive cells was lower in stored slides by an average of 10% (P<0.01). Too few cases in the IHC cohort were FISH positive for any conclusions. HER2 amplification by FISH was unaffected by slide storage. We conclude that use of aged stored slides for proliferation markers TOP2A and Ki67 is feasible but may modestly underestimate true values in endometrial cancer. Pilot studies for particular storage conditions/durations/antigens to be used in translational studies are warranted.