Paraffin-embedded Tissue Blocks Research Articles

SARCOMATOID UROTHELIAL CARCINOMA OF BLADDER - A RARE HISTOLOGICAL VARIANT

Introduction: Urothelial carcinoma (UC) is the most common histological type of bladder cancer. Around 75% of bladder cancers are classified as pure UC, and 25% are urothelial and nonurothelial histological variants. Sarcomatoid UC is a rare variant, accounts for 0.1%–0.3% of all urothelial carcinomas of bladder. It is a biphasic malignant neoplasm with morphologic and immunohistochemical evidence of both epithelial and mesenchymal differentiation. Objective: To study the histomorphological and immunohistochemical features of sarcomatoid urothelial carcinoma. Methods: We received TURBT (Trans Urethral Resection of Bladder Tumor) specimen for histopathological evaluation in the department of pathology, DY Patil medical college, Kolhapur. Routine tissue processing of bits was done and H & E stained sections were subjected for detailed microscopic evaluation and various immunohistochemical markers were applied on the paraffin embedded tissue blocks. Results: A95years old female patient presented with hematuria and anemia.On cystoscopy polypoidal lesion seen in the lateral wall of urinary bladder.No history of previous surgery on urinary bladder.TURBT done. On gross, there were multiple, irregular, grey white strips of tissue altogether weighing 160gms. On microscopy, neoplasm with papillary architecture was seen invading lamina propria. Chondromyxoid, rhabdoid and malignant fibrohistiocytic tumor like areas noted. Immunohistochemical workup revealed positive staining with cytokeratin, vimentin and GATA3 and negative with H-Caldesmon, SMA and Myogenin. Conclusion: Sarcomatoid UC is seen in elderly. Morphologically, the lesion mimics sarcoma, immunohistochemistry workup aids in definitive diagnosis, as it has poor prognosis compared to pure UC.

Deeper sections reveal residual tumor cells in rectal cancer specimens diagnosed with pathological complete response following neoadjuvant treatment.

Guidelines and requirements for diagnosing pathological complete response (pCR) in rectal adenocarcinoma following neoadjuvant treatment vary, and there is currently no consensus on the appropriate number of sections to examine per formalin-fixed, paraffin-embedded (FFPE) tissue block. The consequences of systematic use of deeper sections on the diagnostic accuracy and prognosis for patients classified as ypT0 rectal cancer were investigated. In this retrospective study, 23 out of 155 patients who underwent neoadjuvant therapy and surgical resection between 2015 and 2020 were diagnosed with ypT0 rectal cancer. Three additional deeper sections were cut from each FFPE block from the primary tumor site and reviewed for presence of residual tumor cells. Additional sections revealed residual viable tumor cells in seven patients (30.4%), reducing the rate of pCR in the cohort from 14.8 to 10.3%. Of the seven patients, three patients later had local recurrence or distant metastasis during the follow-up period, compared with one patient with no residual tumor cells in deeper sections (p = 0.07). A nonsignificant reduction in disease-free survival (p = 0.08) was observed in the patients with residual tumor. Systematic use of deeper sections in evaluation of tumor regression in rectal cancer reveals the presence of residual tumor cells in a subset of patients originally diagnosed with pCR based on a single section per FFPE block. Although the results are not statistically significant, it cannot be excluded that accurately distinguishing complete from near-complete response may be clinically relevant for prognostic prediction.

An Immunohistochemical Study on Expression of Laminin5 Gamma2 Chain and Alpha Smooth Muscle actin in Oral Squamous Cell Carcinoma

Background and objectives: Head and neck carcinomas are the biological heterogeneous group of cancers, of which oral cancer is the most common. Ninety percent of oral cancers are squamous cell carcinomas originating from the mucosal epithelium carcinogenesis. Oral squamous cell carcinoma (OSCC) is one of the most prevalent cancers worldwide. The transformation of the normal epithelial cell into a tumor cell depends upon certain changes that happen at the cellular and molecular level which will help in its survival and proliferation. The basement membrane will show invasion by these altered tumor cells into the connective tissue stroma and then their subsequent spread and metastasis, which is an important prognostic indicator. Laminin-5 is a protein which is found to be associated with a migratory phenotype in epithelial neoplastic cells and also the stromal myofibroblasts play a key role in tumor invasion, due to its ability to modify the extracellular matrix. Aim: Laminin-5 (Ln-5) is the major component of the basement membrane and is involved for tumour progression. Alpha-smooth muscle actin (α-SMA) is an isoform of actin, positive in myofibroblasts and is an epithelial to mesenchymal transition (EMT) marker. EMT is a process by which tumor cells develop and are able to metastasize. Tumor cells progression is always followed by alteration in the cell composition and extracellular matrix .Therefore the aim of this study is to detect and evaluate the expression of laminin5 gamma2 and α-SMA in OSCC and to evaluate the association of density of stromal myofibroblasts with tumour budding intensity. Materials and methods: Thirty paraffin embedded tissue blocks of clinically diagnosed and histopathologically confirmed cases of OSCC were evaluated .In this we have divided them into10 well differentiated OSCC,10 moderately differentiated OSCC and 10 poorly differentiated OSCC for laminin-5 and α- smooth muscle actin (SMA) using standard immunohistochemistry technique. ....

TROP-2, Nectin-4, GPNMB, and B7-H3 Are Potentially Therapeutic Targets for Anaplastic Thyroid Carcinoma.

Simple SummaryAnaplastic thyroid carcinoma is a highly aggressive thyroid tumor with a poor prognosis. There are limited choices for the effective treatment of this type of carcinoma. Whether the targets of antibody–drug conjugates are expressed in anaplastic thyroid carcinoma remains unclear. Therefore, we examined expression rates of the following antibody–drug conjugate targets using the tissue microarrays of anaplastic thyroid carcinomas: human epidermal growth factor receptor 2, nectin-4, trophoblast cell surface antigen 2, glycoprotein non-metastatic B, and B7-H3. We found that glycoprotein non-metastatic B and B7-H3 were expressed in most anaplastic thyroid carcinoma tissues. Trophoblast cell surface antigen 2 and nectin-4 were expressed in 65% and 59% of anaplastic thyroid carcinoma tissues, respectively. Trophoblast cell surface antigen 2 was high expressed in anaplastic thyroid carcinoma undifferentiated from papillary thyroid carcinoma. In contrast, nectin-4 expression was high in patients with de novo anaplastic thyroid carcinoma. These cell membrane proteins are potential therapeutic targets for anaplastic thyroid carcinoma.Background: Anaplastic thyroid carcinoma (ATC) is a highly aggressive thyroid tumor with a poor prognosis. However, there are limited choices for ATC treatment. Recently, the effectiveness of antibody–drug conjugates has been demonstrated in various carcinomas. Whether the targets of antibody–drug conjugates are expressed in anaplastic thyroid carcinoma remains unclear. Methods: Fifty-four patients with ATC were enrolled in this study. Tissue microarrays were constructed using the archives of formalin-fixed paraffin-embedded tissue blocks. All sections were stained with the following antibody–drug conjugate targets: human epidermal growth factor receptor 2 (HER2), nectin-4, trophoblast cell surface antigen 2 (TROP-2), glycoprotein non-metastatic B (GPNMB), and B7-H3. Results: HER2 was negative in all tissues, whereas GPNMB and B7-H3 were expressed in most ATC tissues. TROP-2 and nectin-4 were expressed in 65% and 59% of ATC tissues, respectively. TROP-2 was expressed at significantly higher levels in ATC undifferentiated from papillary thyroid carcinoma than in ATC undifferentiated from follicular thyroid carcinoma and de novo ATC. In contrast, nectin-4 expression was markedly higher in patients with de novo ATC than in those with papillary and follicular thyroid carcinoma. Conclusions: TROP-2 and nectin-4 are potential therapeutic targets for ATC undifferentiated from papillary thyroid carcinoma and de novo ATC, respectively. GPNMB and B7-H3 potential for treating all types of ATC.

Evolution of a Paradigm Switch in Diagnosis and Treatment of HPV-Driven Head and Neck Cancer-Striking the Balance Between Toxicity and Cure.

More than a decade after the discovery of p16 immunohistochemistry (IHC) as a surrogate for human papilloma virus (HPV)-driven head and neck squamous cell carcinoma (HNSCC), p16-IHC has become a routinely evaluated biomarker to stratify oropharyngeal squamous cell carcinoma (OPSCC) into a molecularly distinct subtype with favorable clinical prognosis. Clinical trials of treatment de-escalation frequently use combinations of biomarkers (p16-IHC, HPV-RNA in situ hybridization, and amplification of HPV-DNA by PCR) to further improve molecular stratification. Implementation of these methods into clinical routine may be limited in the case of RNA by the low RNA quality of formalin-fixed paraffin-embedded tissue blocks (FFPE) or in the case of DNA by cross contamination with HPV-DNA and false PCR amplification errors. Advanced technological developments such as investigation of tumor mutational landscape (NGS), liquid-biopsies (LBx and cell-free cfDNA), and other blood-based HPV immunity surrogates (antibodies in serum) may provide novel venues to further improve diagnostic uncertainties. Moreover, the value of HPV/p16-IHC outside the oropharynx in HNSCC patients needs to be clarified. With regards to therapy, postoperative (adjuvant) or definitive (primary) radiochemotherapy constitutes cornerstones for curative treatment of HNSCC. Side effects of chemotherapy such as bone-marrow suppression could lead to radiotherapy interruption and may compromise the therapy outcome. Therefore, reduction of chemotherapy or its replacement with targeted anticancer agents holds the promise to further optimize the toxicity profile of systemic treatment. Modern radiotherapy gradually adapts the dose. Higher doses are administered to the visible tumor bulk and positive lymph nodes, while a lower dose is prescribed to locoregional volumes empirically suspected to be invaded by tumor cells. Further attempts for radiotherapy de-escalation may improve acute toxicities, for example, the rates for dysphagia and feeding tube requirement, or ameliorate late toxicities like tissue scars (fibrosis) or dry mouth. The main objective of current de-intensification trials is therefore to reduce acute and/or late treatment-associated toxicity while preserving the favorable clinical outcomes. Deep molecular characterization of HPV-driven HNSCC and radiotherapy interactions with the tumor immune microenvironment may be instructive for the development of next-generation de-escalation strategies.

MicroRNA Profile Associated with Positive Lymph Node Metastasis in Early-Stage Cervical Cancer.

Lymph node metastasis (LNM) is an important prognostic factor in cervical cancer (CC). In early stages, the risk of LNM is approximately 3.7 to 21.7%, and the 5-year overall survival decreases from 80% to 53% when metastatic disease is identified in the lymph nodes. Few reports have analyzed the relationship between miRNA expression and the presence of LNM. The aim of this study was to identify a subset of miRNAs related to LNM in early-stage CC patients. Formalin-fixed paraffin-embedded tissue blocks were collected from patients with early-stage CC treated by radical hysterectomy with lymphadenectomy. We analyzed samples from two groups of patients—one group with LNM and the other without LNM. Global miRNA expression was identified by microarray analysis, and cluster analysis was used to determine a subset of miRNAs associated with LNM. Microarray expression profiling identified a subset of 36 differentially expressed miRNAs in the two groups (fold change (FC) ≥ 1.5 and p < 0.01). We validated the expression of seven miRNAs; miR-487b, miR-29b-2-5p, and miR-195 were underexpressed, and miR-92b-5p, miR-483-5p, miR-4534, and miR-548ac were overexpressed according to the microarray experiments. This signature exhibited prognostic value for identifying early-stage CC patients with LNM. These findings may help detect LNM that cannot be observed in imaging studies.

Immunohistochemical expression of Drosha is reduced in eutopic and ectopic endometrium of women with adenomyosis

The objective of this study was to evaluate the immunohistochemical expression of Dicer, Drosha, and Exportin-5 in the eutopic and ectopic endometrium of women with adenomyosis. Twenty-two paired ectopic and eutopic endometrium from women with adenomyosis and 10 eutopic endometrium samples from control women undergoing hysterectomy were included in the study. Paraffin-embedded tissue blocks were cut and stained for immunohistochemistry. The percentage of epithelial cells positively marked was identified digitally after an automated slide scanning process. Mann-Whitney test or Wilcoxon signed-rank test was performed for independent and paired groups, respectively. A lower expression of Drosha was observed in the eutopic endometrium of women with adenomyosis than in the eutopic endometrium of women without the disease (69.9±3.4% vs 85.2±2.9%, respectively) (P=0.016; 95%CI: 3.4 to 27.4%). We also detected lower Drosha expression in the ectopic endometrium of women with adenomyosis than in the eutopic endometrium of the same women (59.6±3.2% vs 69.9±3.4%, respectively) (P=0.004; 95%CI: 2.3 to 16.7%). Additionally, we observed a correlation between Drosha expression in the ectopic and paired eutopic endometrium (P=0.034, rho=0.454). No significant difference in Dicer or Exportin expression was observed. Predominant pattern of cytoplasmic staining for the anti-Drosha antibody and both a nuclear and cytoplasmic pattern for the anti-Exportin antibody were observed. Drosha expression was significantly lower in the endometrium of women with adenomyosis compared to the eutopic endometrium of asymptomatic women without the disease. Furthermore, its expression was lower in the ectopic endometrium but correlated to the paired eutopic endometrium.

Correlation between Programmed Death Ligand-1(PD-L1) Expression and Driver Gene Mutations in Non-Small Cell Lung Carcinoma- Adenocarcinoma Phenotype.

Background:Targeted therapy in adenocarcinoma is recommended. The use of immune check point inhibitors for the treatment of Non-small cell lung carcinoma (NSCLC) is used as both first-line and the second-line treatment strategy. The current study was undertaken to assess the frequency of programmed cell death ligand-1 (PD-L1) expression with anaplastic lymphoma kinase (ALK), proto-oncogene 1, receptor tyrosine kinase (ROS), epidermal growth factor receptor (EGFR), Kirsten rat sarcoma (KRAS), and v-Raf murine sarcoma viral oncogene homolog B (BRAF)V600E driver gene mutations in NSCLC adenocarcinoma phenotype. It assesses the frequencies of all markers in the cases where both treatment strategies can be implemented. Expression of the all markers was further compared with demographic, clinical parameters, and overall survival rate. Materials and Methods:The formalin-fixed paraffin-embedded (FFPE) tissue blocks were used in immunohistochemistry (IHC) staining and real-time polymerase chain reaction (RT-PCR) for determining the driver genes and PD-L1 expression in the 100 NSCLC-Adenocarcinoma cases. Results:PD-L1 positivity was observed in 26.36% (n=29/110) cases in adenocarcinoma. No significant differences in PD-L1 expression were observed among patients harboring ALK, ROS1, EGFR, KRAS, and BRAF mutations EGFR mutations had significant association with smoking status. (p= 0.008), Thyroid transcription factor 1 (TTF1) (p=0.0005) and Napsin (p=0.002) expression. ALK gene re-arrangement was significantly related to age (p= 0.001), gender (p= 0.009) and smoking status (p= 0.043). The single versus multiple driver mutations were significantly correlated with smoking status (p=0.005). In the survival rate analysis, EGFR (p=0.058), KRAS (p=0.021), and PD-L1 (p=0.039) were significantly high with the positive versus negative group. Conclusions:The current study is a novel attempt to document the co-expression of multiple driver mutations in the NSCLC-adenocarcinoma phenotype. PD-L1 immunopositivity in NSCLC-adenocarcinoma was higher with EGFR mutation as compared to those of KRAS, ALK, ROS, and BRAF driver genes.

Immunohistochemical Expression of Anaplastic Lymphoma Kinase in Invasive Breast Carcinomas- A Retrospective Study

Introduction: Breast carcinoma remains one of the most common causes of mortality among female cancer patients in spite of improvements in treatment modalities. Increased survival rate can be achieved by identification of new targeted therapies. Anaplastic Lymphoma Kinase (ALK) alterations are present in many solid and hematological tumours indicating its role in pathogenesis and treatment. There are studies regarding the expression of ALK in few breast cancers but its importance was not clearly mentioned. Hence, identification of ALK overexpression in breast cancers, particularly in Triple Negative Breast Cancers (TNBC) might play a role in their chemotherapy with the help of ALK inhibitors. Aim: To study the ALK expression in different subtypes of invasive carcinomas of breast. Materials and Methods: This was a retrospective crosssectional study conducted in the Department of Pathology at Great Eastern Medical School (GEMS) &amp; Hospital, Srikakulam, Andhra Pradesh, India, from January 2022 to July 2022. The data of 60 patients, from January 2021 to December 2021 was retrieved using Hospital Information Management System (HIMS) and the Hematoxylin &amp; Eosin-stained slides and formalin fixed paraffin embedded tissue blocks of breast tumour were retrieved and reviewed. Estrogen receptor (ER), Progesterone receptor (PR) and Human Epidermal growth factor Receptor 2 (HER2) immunostains were performed and categorized based on molecular classification as Luminal, Her2 and Triple negative. ALK Staining was performed on all cases and its expression was studied. Statistical Package for Social Sciences (SPSS) software version 2.0 was used for analysis of data. Mean with standard deviation is used for quantitative variables and prevalence, ratio is used for quantitative variables. Chi-square test and Fischer exact test were used for detecting significance. The p-value &lt;0.05 was considered as significant. Results: Out of 60 cases, majority(N=22;36.66%) of patients were in the age group of 51 to 60 years. The mean tumour size was 3.2±0.5 cm. The most common histological type was invasive breast carcinoma, No Special Type (N=44;73.34%). Majority of the tumours showed grade 1 and 2 with 24 (40%) and 25 (41.67%) cases respectively. A total of 30 cases (50%) of tumours belong to stage T2. Luminal molecular subtype is the most common 31 cases (51.67%) followed by TNBC, 16 cases(26.67%) and Her2neu 13 cases (21.67%). Among all the cases, ALK overexpression was seen in 17 (28.33%) cases and among different molecular subtypes, its expression is seen in 5 (8.33%) cases of Luminal type, 3 (5.0%) cases of HER2 type and in 9 (15.0%) cases of TNBC cases. Conclusion: Immunohistochemical analysis showed ALK over expression in a substantial proportion of cases and possibly plays a significant role in aggressive behaviour of breast cancer. ALK inhibitors offer an opportunity to treat aggressive subtypes of breast cancer.

EVALUATION OF NLRP3 INFLAMMASOME PROTEIN EXPRESSION IN ULCERATIVE COLITIS

The aim: This study aimed to evaluate the NLRP3 immunoreactivity in ulcerative colitis in various clinical presentations. Materials and methods: The retrospective study involved 80 formalin fixed paraffin embedded tissue blocks. These were divided into 50 samples of ulcerative colitis and 30 with normal colonoscopy findings. NLRP3 protein expression patterns were evaluated in various cellular components in tissue sections using immunohistochemistry method. Results: NLRP3 inflammasome was significantly expressed with higher percentage among ulcerative colitis tissue sections compared with normal tissue sections. Intense staining was observed in Paneth cells at the base of crypts, inflammatory cells infiltrated between glands and near the base of crypts. Variable intensities of NLRP3 staining were observed in surface epithelial cells and glandular epithelium. Statically higher percentage of expression was found among active disease and patients with extra-intestinal complications. Conclusions: The NLRP3 protein expression pattern was upregulated among various cellular compartments among ulcerative colitis and correlated with disease activity.

Análise imuno-histoquímica da proteína ORF2 e proteína ORF3 do vírus da hepatite E em fígados de suínos no estado de Mato Grosso, Brasil

ABSTRACT: Hepatitis E is an emerging zoonotic disease caused by hepatitis E virus (HEV). Immunohistochemistry (IHC) can be used to verify viral presence in human and swine livers. The aim of this study was to comparatively analyze the immunolabeling of the ORF2 protein (pORF2) versus the ORF3 protein (pORF3) of HEV in swine livers from subsistence farms in the state of Mato Grosso, Brazil. This study included 25 liver samples formalin fixed paraffin embedded tissue block from a published molecular detection and immunohistochemistry (IHC) study, which used the HEV pORF3 protein, demonstrating 4% (1/25) of positive immunolabeling and 96% (24/25) negative, in contrast to the molecular exam that showed 24% (6/25) of liver samples positive and 76% (19/25) negative. In order to increase the sensitivity of the IHC technique, these samples were analyzed using the antibody for the detection of HEV pORF2, showing 24% (6/25) immunolabeling positive and 76% (19/25) negative, equivalent to the result of molecular analysis on corresponding samples. Thus, the use of antibody to pORF2 increased the number of HEV cases detectable in the IHC by 600%. The IHC added to molecular techniques can be used as a tool for monitoring viral presence in swine livers, constituting a sensitive diagnostic methodology when liver samples fixed in formalin and embedded in paraffin are available.

The impact of different coverslipping methods in the quality of the whole slide images used for diagnosis in pathology

Digital pathology workflow aims to create whole slide images (WSIs) for diagnosis. The quality of the WSIs depends primarily on the quality of the glass slides produced by the pathology laboratory, where the coverslipping method plays an important role. In this study we compare the glass, the film, and the liquid coverslipping methods to evaluate which ones are suitable to create WSIs for diagnosis. The study included 18 formalin-fixed paraffin-embedded tissue blocks. Of each block, 3 consecutive sections were covered using 1 of the 3 methods. The slides were scanned and evaluated for quality criteria by 2 pathologists experienced in digital pathology. The coverslipping method interferes with the quality of the WSIs, as well as with the scanning time and the file size of the WSIs. All coverslipping methods were found suitable for diagnosis. The glass and liquid methods were manual and had similar results concerning the presence of air bubbles/polymer accumulation, air drying artefacts, tissue exposed, and staining alterations. The glass method was the one with more air bubbles. The liquid method was associated with more alterations on the WSIs, but with the lowest file sizes. Automation of coverslipping and calibration of the scanner for the coverslipping method chosen by the pathology laboratory are relevant for the final quality of the WSIs.

Evaluation of immunohistochemical expression of stem cell markers (NANOG and CD133) in normal, hyperplastic, and malignant endometrium.

Cancer stem cells (CSC) are a potential cause for recurrence, metastasis, and resistance of tumors to different therapeutic modalities like hormonal radiotherapy and chemotherapy. We investigated two CSC markers (NANOG and CD 133) in normal, hyperplastic endometrium and endometrial carcinoma. A total of 93 formalin-fixed paraffin-embedded tissue blocks were used for immunohistochemical expression of NANOG and CD133 markers. NANOG expression was detected in 88.37% of endometrial carcinoma cases compared to 15% of the normal proliferative endometrium and 60% of hyperplasia cases. In endometrial carcinoma, high NANOG expression was significantly correlated with high grade, deep myometrial invasion, lymph node metastasis, and high stage with p-values (0.009, 0.005, 0.014, and 0.003, respectively). CD133 was positive in 76.74% of endometrial carcinoma cases, and it showed a significant correlation with deep myometrial invasion, positive lymph node, positive lymphovascular invasion, and high stage (p-values 0.003, 0.001, 0.003, and 0.013, respectively). Normal endometrium showed less expression of CD133 (only 5%) than hyperplasia and endometrial carcinoma with a statistically highly significant difference (p less than 0.0001). Hyperplastic cases with atypia expressed higher CD133 than those without atypia (6 out of 12 versus 3 out of 18). However, this difference was not statistically significant (p-value 0.111). The cancer stem cell markers NANOG and CD 133 are expressed in a high percentage in endometrial carcinoma compared to normal and hyperplasia and their expression is positively correlated with the aggressive behavior of the tumor. High expression of these two markers in apparently normal tissue around the tumor and in hyperplastic conditions with atypia suggests the possibility to use NANOG and CD133 expression as a diagnostic marker distinguishing dysplasia from reactive atypia. Therefore, inhibition of these markers can be a promising method to stop the progression of early cancers.

Detection of Human PapillomaVirus and its Association with Potentially Malignant Disorders and Oral Squamous Cell Carcinoma: A Retrospective Study.

Aim:This study was carried out with the objective of recognizing the involvement of human papillomavirus (HPV) in conditions of oral squamous cell carcinoma (OSCC) and oral potentially malignant conditions and lesions.Materials and Methods:OSCC samples from thirty patients were collected and thirty paraffin-embedded, oral premalignant specimens were collected. It was decided to use as a negative control samples gingiva from healthy individuals who had no prior history of oral proliferative leukoplakia or OSCC. The samples were then processed using a paraffin-embedded tissue block method. Deoxyribonucleic acid will be extracted from the paraffin-embedded tumor sample of sixty case subjects and tested for the presence of HPV DNA using polymerase chain reaction.Results:In study Group 1, 24 samples were taken from males and 6 from females. In study Group 2, 26 cases were males and 4 were females. In Group 1, 16 cases of moderately differentiated and 14 cases of well differentiated each were present. No case of poorly differentiated were found. There was statistically no significant difference present between male and female when Group 1 and Group 2 was considered. In study Group 1, 0 samples had positive HPV DNA detection, whereas 30 samples had negative HPV DNA detection. In study Group 2, 0 samples had positive HPV DNA detection, whereas 30 samples had negative HPV expression.Conclusion:According to our findings, certain incidences of OSCC may be linked to HPV, however, this was not the case for all of the cancers.

Histopathological Study of Carcinoma Cervix through Reviewing of Female Genital Tract Cancer Cases of Pathology Archive, Benghazi University from (2002-2011)

Female genital tract (FGT) is the most common site for tumors in females. The most common type of (FGT) cancers is cervical, ovarian and endometrial carcinoma. There are other less common tumor's including tumors of the vagina, vulva and fallopian tubes. Cervical cancer is the second most commonly diagnosed cancer worldwide and the fourth leading cause of cancer death in women. To determine the frequency of (FGT) malignancy found at the pathology archive of the Pathology Department at the Faculty of Medicine in the University of Benghazi between the years 2002-2011. Also to define the proportion of cervix cancer cases as compared with other (FGT) that were recorded in the archive. Moreover, to study the histopathological type of cervical cancer, the age groups and at which grades of differentiation are mostly determined. Retrospective histopathological analysis of (FGT) 330 cancer cases diagnosed between (2002-2011). During the reviewing of (FGT) cancer cases of pathology archive all cases of cervical cancer between 2002-2011 were collected and all formalin-fixed paraffin-embedded (FFPE) tissue blocks of the patients with cervical cancer in that period were collected, and processed to stained slides. In an analysis of 330 FGT cancer cases registered in the pathology archive. (52%) were carcinoma of the endometrium and this was the commonest. This was followed by the carcinoma of the cervix (30.9%), ovarian tumors (13%), vaginal tumors (2%), fallopian tumors and vulva tumors (1%). The most common type of carcinoma cervix was squamous cell carcinoma (62%), adenocarcinoma was (31%) and the least common type was adenosquamous carcinoma (7%). The mean age of patients was 54.67 years. Most cases presented at the postmenopausal period (62.5%). Most cases were poorly differentiated (47.1%), followed by moderately differentiated (33.3%) and well-differentiated (17.6%).

MiR-515-3p, miR-623, miR-1272 and Notch3 protein as new biomarkers of Hepatocellular carcinoma.

Hepatocellular carcinoma (HCC) is a diversity of hepatocellular neoplasms and is more prevalence in people with chronic liver disease and cirrhosis. It has been revealed that modification in miRNA regulation possibly will be elaborated in HCC pathogenesis. In this research 40 samples of HCC subjects and 40 samples of healthy liver were considered. Total RNA was obtained from paraffin-embedded tissue blocks and miR-515, miR-623 and miR-1272 gene expression levels were quantified by Real-Time Quantitative Reverse Transcription PCR. Likewise, the Notch protein quantity was assayed in ffpe materials by immunohistochemistry. Our study disclosed that Notch protein deals was ominously elevated in cancer cells than healthy cells (p<0.05). Data analysis also displayed that miR-515, miR-623 and miR-1272 expression levels were 3.8, 4.7, and 2.9 fold in normal tissues, respectively (p<0.05). Furthermore, it was found that expression levels of these genes are not dependent by hepatitis B and hepatic cirrhosis and it could be used as a marker of high specificity and sensitivity for the diagnosis of HCC. Our study demonstrated main role of miR-515, miR-623 and miR-1272 in HCC pathogenesis and similarly disclosed that these genes expression could be utilized in HCC prognosis.

Histomorphometric evaluation of human extraction sockets treated with autologous fibrin, sticky bone or biphasic calcium phosphate.

It is essential to maintain the alveolar bone ridge to ensure the success of implant therapy. Platelet-rich fibrin (PRF) may benefit bone repair, but its quantitative microscopic results are still inconclusive. The aim of this study was to histomorphometrically analyze human dental alveoli after extraction treated with autologous fibrin, biphasic calcium phosphate or sticky bone. The sample consisted of healthy adult volunteer patients, with clinical and tomographic indication for single post-extraction graft of upper premolars for maintenance of the alveolar ridge and subsequent implantation. The 10 remaining patients in the study were divided into three groups according to the type of filling used in the dental socket: autologous PRF plug covered by a PRF membrane (G1), PRF associated with an alloplastic graft based on hydroxyapatite with beta tricalcium phosphate covered by a collagen membrane (G2) or alloplastic graft based on beta tricalcium phosphate covered by collagen membrane (control). After 8 months, bone biopsies were performed at the grafted sites and the patients underwent implant-prosthetic rehabilitation. Paraffin-embedded tissue blocks were routinely processed and sectionsfrom different depths were mounted in 3 slides and stained with HE. The histomorphometric evaluation analyzed 30 photomicrographs per block, quantifying the percentage presence of newly formed bone, connective tissue and remaining biomaterial using the ImageJ software. Parametric data enabled intergroup comparisons using ANOVA and Tukey's post-hoc test for multiple comparison with statistical significance of 5% (p<0.05), with normality of the 3 groups by the Jarque-Bera test (p>0.05). There was a higher mean of newly formed bone in G1 (68.83%) compared to G2 (35.69%) and control (16.28%). There was greater presence of connective tissue in the control (61.56%). Remaining biomaterial was higher in G2 (15.75%), but did not differ statistically from the control. Bone regeneration obtained with PRF alone or sticky bone suggests the efficacy of these therapies, encouraging the clinical use of this blood concentrate in dental procedures.

Micro-computed tomography: A novel diagnostic technique for the evaluation of gastrointestinal specimens.

Micro-computed tomography (micro-CT) is a non-destructive modality that can be used to obtain high-resolution three-dimensional (3 D) images of the whole sample tissue; the usefulness of micro-CT has been reported for evaluation of breast cancer and lung cancer. However, this novel diagnostic technique has never been used for evaluating endoscopically resected gastrointestinal specimens. In the present study, we scanned 13 formalin-fixed paraffin-embedded (FFPE) tissue blocks of a normal human colon and gastric tissue samples using micro-CT. The evaluation comprised a comparison of the acquired whole block images with the images of the corresponding cross-sectional slice of the hematoxylin and eosin-stained slide. Micro-CT was able to produce images of the whole sample and clearly depict tissues such as glandular structures, muscularis mucosae, and blood vessels in the FFPE tissue blocks of normal gastrointestinal samples. Furthermore, the 3 D reconstructed could be used to create a cross-sectional image and reflected the surface structure of samples obtained from any site. Micro-CT has the potential to become a highly promising pathological diagnostic assistance tool for endoscopically resected gastrointestinal specimens in combination with conventional microscopic examination.

Detection of Epstein Barr Virus (EBV) Antigen in Oral Squamous Cell Carcinoma (OSCC) by Immunohistochemistry in Patients from KPK Province of Pakistan.

Objective: To determine the frequency of EBVLMP-1 antigen positivity in OSCC and normal oral squamous epithelial tissues (control) by immunohistochemistry (IHC) and to see the effect of age and gender in both the OSCC and control tissues. Study Design: Descriptive, Cross Sectional, Multicenter study. Setting: Pathology/ Peshawar Medical College (PMC), Peshawar Dental College and Sardar Begum Dental College, Peshawar. Period: April 2018 to September 2018. Material &amp; Methods: Conducted on total 60 samples divided into two groups. Group A: 30 formalin fixed paraffin embedded tissue blocks of OSCC cases and Group B: 30 non neoplastic oral epithelial tissues (control). Both groups A and B slides were stained through immunohistochemistry for EBV LMP-1 monoclonal antigen. Results: 30 cases of OSCC Group A and 30 cases of non-neoplastic oral mucosa Group B were selected for the LMP-1 staining by immunohistochemistry. The mean age of OSCC was 56.5 years (±14.47 Standard deviation) with a range of 20-80 years. Male: Female ratio was 1.3:1 .OSCC commonly involved buccal mucosa. 80% (n=24/30) OSCC cases were grade 1, all the cases of OSCC (100%) showed epithelial positivity for EBV LMP-1 antigen, whole (29/30) cases of non-neoplastic oral mucosa cases showed positivity and there is no statistically significant difference in both Groups A and B. Conclusion: With the findings of EBV positivity in both the OSCC and all of the control cases; it is concluded that EBV does not play an important role in etiology of OSCC. This suggests that the infection by this ubiquitous virus (EBV) occurs at some point in the life of a person leaving LMP, protein latent in the cells of oral epithelium.

Genetic and epigenetic profiling of the BRCA1 / 2 genes in solitary ovarian cancer and multiple primary ovarian tumors

Background. Ovarian cancer is a complex and poorly studied disease that kills nearly 70–80 % of patients. Therefore, practitioners are interested in any opportunity of improving survival of these patients. From this point of view, investigation of genetic and epigenetic functions associated with this pathology is quite promising.Objective: to assess clinical and morphological characteristics of tumors in ovarian cancer patients, considering the presence of mutations and methylation in the BRCA1/2 gene.Materials and methods. This study included 180 ovarian cancer patients (FIGO stage I–IV) treated in the N. N. Blokhin Russian Cancer Research Center between 2008 and 2019. Study participants were divided into 3 groups according to their BRCA status and the number of primary tumors. We collected and analyzed venous blood, biopsy samples of ovarian cancer, archived histological sections, and paraffin-embedded tissue blocks. DNA isolated from venous blood was used to identify the following germline mutation by pyrosequencing: BRCA1 5382insC, BRCA1 4153delA, BRCA1 185delAG, and BRCA26174delT. DNA isolated from biopsy specimens and paraffin-embedded tissue specimens was used to analyze methylation in the promoter regions of the BRCA1 and BRCA2 genes by bisulfite sequencing (PyroMark Q24 DNA Sequencer; Qiagen, USA) with specific primers targeting promoter regions of the BRCA1 and BRCA2 genes.Results. Molecular testing demonstrated that the frequency of BRCA1 gene mutations was 21.1 % (38/148) in patients with solitary ovarian cancer and 40.6 % (13/32) in patients with multiple primary ovarian cancers. The frequency of methylation of the BRCA1 gene promoter was 2.2 % (18/148) in patients with solitary ovarian cancer and 3.1 % (1 case) in patients with multiple primary ovarian cancers. All BRCA1 methylated ovarian tumors were serous adenocarcinomas, including high grade tumors in 15 patients (78.9 %) and low-grade tumors in 4 patients (21.1 %).Conclusion. Hypermethylation of the BRCA1 gene promoter was observed only in individuals with sporadic serous ovarian cancer. No methylation was detected in patients with non-serous ovarian cancer, as well as in patients carrying BRCA1 gene mutations (both with solitary ovarian cancer and with primary multiple ovarian tumors).