Granular cell tumor of the breast: a case report and review of literature

A 22-year-old female patient presented with a breast mass lesion with a clinical suspicion of a fibroadenoma. Histological evaluation revealed a rare benign neoplasm - granular cell tumor

Bcl-2 and Ki67 as specific prognostic markers in estrogen receptor positive breast cancers

Objective: Breast cancer is a heterogeneous disease, so many biomarkers can be potential predictors in its outcome. The aim of this study is to test the hypothesis that combinatorial assessment of both Ki-67 proliferation index and B-cell Lymphoma 2 (Bcl-2) protein, would provide prognostic information on occurrence of relapses in breast cancer patients. Method: Immunohistochemical expression of Ki67 and Bcl-2, represented as Ki67/Bcl-2 index, were evaluated in 183 Estrogen Receptor positive breast cancer patients from 2007 to 2012, compared with other clinical-pathologic findings. During the follow up period (45–114 months) recurrences were observed in 36 patients (19.7 %). Results: A significant correlations were notified between Ki67/Bcl-2 index with age, tumour size, nuclear grade, histologic grade, lympho-vascular invasion, progesterone receptor status and expression of p53 protein product (p < 0,05). The occurrence of relapses in the group of low Ki67/high Bcl-2 index was lower, compared with the group of high Ki67/low Bcl-2 index breast cancer patients (p < 0.01). Conclusion: The combination of Ki67 and Bcl-2 biomarkers is useful tool in prediction of relapses in Estrogen positive breast cancer patients

Evaluation of PD-L1 expression in various formalin-fixed paraffin embedded tumour tissue samples using SP263, SP142 and QR1 antibody clones

Background & objectives: Cancer cells can avoid immune destruction through the inhibitory ligand PD-L1. PD-1 is a surface cell receptor, part of the immunoglobulin family. Its ligand PD-L1 is expressed by tumour cells and stromal tumour infltrating lymphocytes (TIL). Methods: Forty-four cancer cases were included in this study (24 triple-negative breast cancers (TNBC), 10 non-small cell lung cancer (NSCLC) and 10 malignant melanoma cases). Three clones of monoclonal primary antibodies were compared: QR1 (Quartett), SP 142 and SP263 (Ventana). For visualization, ultraView Universal DAB Detection Kit from Ventana was used on an automated platform for immunohistochemical staining Ventana BenchMark GX. Results: Comparing the sensitivity of two different clones on same tissue samples from TNBC, we found that the QR1 clone gave higher percentage of positive cells than clone SP142, but there was no statistically significant difference. Comparing the sensitivity of two different clones on same tissue samples from malignant melanoma, the SP263 clone gave higher percentage of positive cells than the QR1 clone, but again the difference was not statistically significant. Comparing the sensitivity of two different clones on same tissue samples from NSCLC, we found higher percentage of positive cells using the QR1 clone in comparison with the SP142 clone, but once again, the difference was not statistically significant. Conclusion: The three different antibody clones from two manufacturers Ventana and Quartett, gave comparable results with no statistically significant difference in staining intensity/ percentage of positive tumour and/or immune cells. Therefore, different PD-L1 clones from different manufacturers can potentially be used to evaluate the PD- L1 status in different tumour tissues. Due to the serious implications of the PD-L1 analysis in further treatment decisions for cancer patients, every antibody clone, staining protocol and evaluation process should be carefully and meticulously validated

Androgen Receptor Expression in Epithelial and Stromal Cells of Prostatic Carcinoma and Benign Prostatic Hyperplasia

BACKGROUND: Prostatic carcinoma (PCa) derives from prostatic epithelial cells. However stromal microenvironment, associated with malignant epithelium, also plays a role in prostatic carcinogenesis. Alterations in prostatic stromal cells contribute to the loss of growth control in epithelial cells that lead to progression of PCa.AIM: To analyse the differences between Androgen Receptor (AR) expression in both epithelial and stromal cells in PCa and the surrounding benign prostatic hyperplasia (BPH) and to compare the results with tumour grade.MATERIAL AND METHODS: Samples from 70 cases of radical prostatectomy specimens were used. The expression and intensity of the signal for AR was analysed in the epithelial and stromal cells of PCa and BPH, and the data was quantified using histological score (H-score).RESULTS: AR showed significantly lower expression in both epithelial and stromal cells of PCa compared to BPH. In PCa a significant positive correlation of AR expression was found between stromal and epithelial cells of PCa. AR expression showed a correlation between the stromal cells of PCa and tumour grade.CONCLUSION: AR expression is reduced in epithelial and stromal cells of PCa. Expression of AR in stromal cells of PCa significantly correlates with tumour grade

Y-chromosome haplogroup architecture confers susceptibility to azoospermia factor c microrearrangements: a retrospective study

Aim To assess the association between azoospermia factor c microrearrangements and semen quality, and between Y-chromosome background with distinct azoospermia factor c microrearrangements and semen quality impairment. Methods This retrospective study, carried out in the Research Center for Genetic Engineering and Biotechnology “Georgi D. Efremov,” involved 486 men from different ethnic backgrounds referred for couple infertility from 2002- 2017: 338 were azoospermic/oligozoospermic and 148 were normozoospermic. The azoospermia factor c microrearrangements were analyzed with sequence tagged site and sequence family variant markers, quantitative fluorescent polymerase chain reaction, and multiplex ligation probe amplification analysis. The Y-haplogroups of all participants were determined with direct single nucleotide polymorphism typing and indirect prediction with short tandem repeat markers.Results Our participants had two types of microdeletions: gr/gr and b2/b3; three microduplications: b2/b4, gr/gr, and b2/b3; and one complex rearrangement gr/gr deletion + b2/b4 duplication. Impaired semen quality was not associated with microrearrangements, but b2/b4 and gr/ gr duplications were significantly associated with haplogroup R1a (P < 0.001 and P = 0.003, respectively) and b2/b3 deletions with haplogroup E (P = 0.005). There were significantly more b2/b4 duplication carriers in Albanians than in Macedonians with haplogroup R1a (P = 0.031). Conclusion Even though azoospermia factor c partial deletions/duplications and Y-haplogroups were not associated with impaired semen quality, specific deletions/ duplications were significantly associated with distinct haplogroups, implying that the Y chromosome background may confer susceptibility to azoospermia factor c microrearrangements

Comparative proteomics analysis of human FFPE testicular tissues reveals new candidate biomarkers for distinction among azoospermia types and subtypes.

Understanding molecular mechanisms that underpin azoospermia and discovery of biomarkers that could enable reliable, non-invasive diagnosis are highly needed. Using label-free data-independent LC-MS/MS acquisition coupled with ion mobility, we compared the FFPE testicular proteome of patients with obstructive (OA) and non-obstructive azoospermia (NOA) subtypes hypospermatogenesis (Hyp) and Sertoli cell-only syndrome (SCO). Out of 2044 proteins identified based on ≥2 peptides, 61 proteins had the power to quantitatively discriminate OA from NOA and 30 to quantitatively discriminate SCO from Hyp and OA. Among these, H1-6, RANBP1 and TKTL2 showed superior potential for quantitative discrimination among OA, Hyp and SCO. Integrin signaling pathway, adherens junction, planar cell polarity/convergent extension pathway and Dectin-1 mediated noncanonical NF-kB signaling were significantly associated with the proteins that could discriminate OA from NOA. Comparison with 2 transcriptome datasets revealed 278 and 55 co-differentially expressed proteins/genes with statistically significant positive correlation. Gene expression analysis by qPCR of 6 genes (H1-6, RANBP1, TKTL2, TKTL1, H2BC1, and ACTL7B) with the highest discriminatory power on protein level and the same regulation trend with transcriptomic datasets, confirmed the proteomics results. In summary, our results suggest some underlying pathways in azoospermia and broaden the range of potential novel candidates for diagnosis. SIGNIFICANCE: Using a comparative proteomics approach on testicular tissue we have identified several pathways associated with azoospermia and a number of testis-specific and germ cell-specific proteins that have the potential to pinpoint the type of spermatogenesis failure. Furthermore, comparison with transcriptomics datasets based on genome-wide gene expression analyses of human testis specimens from azoospermia patients identified proteins that could discriminate between obstructive and non-obstructive azoospermia subtypes on both protein and mRNA levels. Up to our knowledge, this is the first integrated comparative analysis of proteomics and transcriptomics data from testicular tissues. We believe that the data from our study contributes significantly to increase the knowledge of molecular mechanisms of azoospermia and pave the way for new investigations in regards to non-invasive diagnosis

Clinicopathological and histomorphological association in K-ras mutated colorectal cancer

Background & objectives: KRAS mutation is frequently identified in advanced colorectal carcinoma (CRC)and its prognostic significance with histological features have remained to be clarified. The aim of this study is to evaluate the clinicopathological and histomorphological characteristics in K-ras mutated patients with CRC. Methods: In this retrospective study, 420 CRC patients who underwent surgical treatment in our hospital (January 2018- December 20200 have been included. K-ras mutation testing was performed in 265 patients, detected by Cobas K-ras mutation kit to identify frequent mutations (codons 12,13,61). Clinicopathological and histomorphological data were compared with the K-ras status and correlations were evaluating using Pearson’s Chi-square test. Results: K-ras mutations were found in 148 patients (39,2%), frequently identified in older males, and in advanced stages of the disease. There was association of the K-ras mutation with the degree of tumour differentiation (G3), tumour necrosis and inflammatory response of the tumour tissue (p<0.05). No association was found between the mutational status and the tumour extention, localization, lymphonodal status and tumour type. Conclusion: According to the certain limitations of this retrospective study using a single detection kit that include common codon changes in K-ras gene it is obvious that further studies on the histological results and their prognostic value of rare KRAS codon variants are necessary. From the other perspective, the present study demonstrated a moderate association between KRAS-mutated CRCs and specific histology, and, to a certain degree, an association between histology and prognosis, according to KRAS mutation status

Anaplastic lymphoma kinase (ALK) expression in lung adenocarci-noma - clinicopathologic and morphologic features

Background & Objectives:The aim of this study is to explore the statusof ALK in lung adenocarcinomas and to evaluate the relationship be-tween ALK expression and clinicopathological and morphological fea-tures of the primary tumour.Methods:In total, 86 cytological and biopsy specimens were tested forALK status using automated immunostainer with Ventana anti-ALK(D5F3) antibody. The expression of ALK was correlated with patient’sage, sex and histological features of the primary tumour.Results:ALK positive expression was detected in 15 (17,4%) cases, insignificantly younger age group than ALK negative patients (p<0.05).Mucinous type of lung adenocarcinomas were predominantly ALK pos-itive (43,7%), followed by papillary type (25%), acinic type (13%) andsolid type (3%). There was strong correlation between the ALK expres-sion in surgical biopsy and cytological specimens (p<0.05).Conclusion:These results represent that immunohistochemical expres-sion of ALK gene rearrangement is valid detection technique revealingdistinctive clinicopathological and morphological features of the tumourespecially in scant biopsies and cytological specimens

Rapid detection of mismatch repair proteins by immunohistochemistry in colorectal cancer patients

Background & Objectives:Lynch syndrome is an inherited disorder thatincreases the risk of many types of cancer, particularly colorectal andendometrial cancer. Therefore, all newly diagnosed colorectal cancersshould be screened for Lynch syndrome. New immunohistochemistry(IHC) based tests which detect mismatch repair (MMR) proteins offerquick and reliable identification ofpatients with probable Lynchsyndrome.Methods:In this prospective study, we evaluated 50 cases of colorectalcancer patients using the Ventana MMR IHC Panel. The panel containsfive primary mouse or rabbit monoclonal antibodies: MLH-1 (M1),PMS2 (EPR3947), MSH2 (G219-1129), MSH6 (44) and BRAF V600E(VE1). The analysis was performed on automated platform VentanaBench Mark GX, using 4μthin tissue sections from representative tu-mour tissue paraffin blocks.Results:Of the 50 analysed cases, 3 cases showed absence of positivityfor MLH1 and PMS2 markers. Two of these cases had MLH1 promoterhypermethylation and were classified as sporadic cancers. One case wasnegative for PMS2 marker and one case was negative for MSH6 marker.In total, 3 of the 50 cases analysed were sent for further Lynch syndrometesting. Two of these three patients were female and also had a history ofendometrial cancer prior to the diagnosis of colorectal cancer. One casewas positive for BRAF V600E antibody.Conclusion:Immunohistochemical detection of MMR proteins enablesquick detection of patient with probable Lynch syndrome. Further iden-tification of the syndrome in patients and family members may result inearly detection and possible cancer prevention in these patients

The Breast cancer Imunophenotypes in Correlation with Classical Morphological Parameters as a Predictive Model for Recurrenec Risk

Objective: Breast cancer is the most commonly diagnosed cancer in women worldwide characterized by molecular and clinical heterogeneity that results with multiple intrinsic tumor subtypes. The aim of this study was to evaluate the occurrence of relapses in the different immunophenotypes of breast cancer (BC) associated with different histomorphological parameters. Methods: The retrospective population study included 173 breast cancer patients diagnosed between 2007 and 2010 in our hospital. Molecular subtype classification was performed on immunohistochemical surrogates for estrogen (ER) and progesterone receptor (PR), as well as for proliferation index determined with Ki- 67 antibody and Human Epidermal Growth Factor receptor 2 (HER-2), according to St. Gallen International Expert Consensus recommendations from 2013. During the follow-up period (min.12,2, max. 75,3, mean 46,6+16,6 months), recurrences were observed in 35 (20,2%). BC immunophenotypes and classic histomorphological and clinical parameters were analyzed in terms of disease free survival (DFS) in a multivariate fashion using a Cox regression model. Results: Our results showed that proportions of breast cancer immunophenotypes were: Luminal A-26,56%; Luminal B-41.67%; HER2+ 18,75% and Triple-negative- 13,02%. In the Univariate analyses there was a significant difference in the distribution of age, tumor diameter, mitotic index, lympho-nodal ratio, Nottingham Prognostic Index (NPI), stage of the disease, Ki67 PI and the bcl-2 overexpression at the diagnosis among the four BC immunophenotypes. In the the multivariate analyses, the age of the patients, the tumor diameter and the stage of the disease were represented as independent prognostic factors of recurrent disease in different BC immunophenotypes. Conclusion: The prognostic value of breast cancer immunophenotypes persists when adjusting the age, the tumor diameter and the stage of the disease, as clinical parameters. This “morphologic-molecular” model was robust in relapse prediction and recurrence risk stratified by traditional prognostic parameters