Histological and Somatic Mutational Profiles of Mismatch Repair Deficient Endometrial Tumours of Different Aetiologies

Background: Mismatch repair deficient (MMRd) tumours may arise from somatic events acquired during carcinogenesis or in the context of Lynch syndrome (LS), an inherited cancer predisposition condition caused by germline MMR pathogenic variants. Our aim was to explore whether sporadic and hereditary MMRd endometrial cancers (EC) display distinctive tumour biology. Methods: Clinically annotated LS-EC were collected. Histological slide review was performed centrally by two specialist gynaecological pathologists. Mutational analysis was by a bespoke 75- gene next-generation sequencing panel. Comparisons were made with sporadic MMRd EC. Multiple correspondence analysis was used to explore similarities and differences between the cohorts. Results: After exclusions, 135 LS-EC underwent independent histological review, and 64 underwent mutational analysis. Comparisons were made with 59 sporadic MMRd EC. Most tumours were of endometrioid histological subtype (92% LS-EC and 100% sporadic MMRd EC, respectively, p = NS). Sporadic MMRd tumours had significantly fewer tumour infiltrating lymphocytes (p ≤ 0.0001) and showed more squamous/mucinous differentiation than LS-EC (p = 0.04/p = 0.05). PTEN mutations were found in 88% sporadic MMRd and 61% LS-EC, respectively (p < 0.001). Sporadic MMRd tumours had significantly more mutations in PDGFRA, ALK, IDH1, CARD11, CIC, MED12, CCND1, PTPN11, RB1 and KRAS, while LS-EC showed more mutations affecting SMAD4 and ARAF. LS-EC showed a propensity for TGF-β signalling disruption. Cluster analysis found that wild type PTEN associates predominantly with LS-EC, whilst co-occurring mutations in PTEN, PIK3CA and KRAS predict sporadic MMRd EC. Conclusions: Whilst MMRd EC of hereditary and sporadic aetiology may be difficult to distinguish by histology alone, differences in infiltrating immune cell counts and mutational profile may predict heterogenous responses to novel targeted therapies and warrant further study.

Digital pathology in the immunohistochemical evaluation of biomarkers in breast cancer

Background Currently, the evaluation of biomarkers HER-2 and ER is critical for targeting therapy for breast cancer and is performed by experienced pathologists, which takes time and causes a certain interobserver variability. The use of histological slide scanners coupled with the application of software for immunoexpression quantification can be profitable in pathological laboratories routine. This study aims to compare the quantification of HER-2 and ER immunoexpression employing automatic algorithms, using as standard the evaluation performed by a pathologist. Methods From the archive since 2016 to 2018 were retrieved 75 immunostained slides for HER-2 and 76 immunostained slides for ER with invasive breast carcinoma diagnosis. The slides were scanned in Aperio CS2 and the immunostaining signal was quantified in Aperio Imagescope using a membrane and nuclear algorithm, respectively. The concordance between the scores obtained and the previous assessment was calculated by Cohen’s Kappa coefficient as well as the sensitivity and specificity for each algorithm. Results Both algorithms showed an almost perfect concordance with the conventional method (k = 0.94 for HER-2; k = 0.92 for ER). Digital evaluations presented a sensitivity of 100% for both biomarkers, and a specificity of 100% for HER-2 and 80% for ER. Conclusions This assay showed that applying signal quantification software for HER-2 and ER in digitized slides is accurate, as these tools have potential to be implemented in laboratory routine. However, it will be necessary to increase the sample to obtain more reliable values and extend this study to the PR and Ki67 biomarkers since both carry prognostic and predictive information as well.

The Effects of Brotowali (Tinospora crispa) Extract on The Number of Macrophage Cells in Wistar Rats (Rattus norvegicus) Suffering from Periodontitis

Background: Periodontitis is an inflammatory disease of the periodontal tissues usually provoked by Actinobacillus actinomycetemcomitans (A. actinomycetemcomitans) bacteria. In the periodontitis healing process, macrophages play a significant role in the inflammatory stage. Brotowali (Tinospora crispa) consists of flavonoids that can stimulate periodontitis healing. Methods: Brotowali extract was derived from dried brotowali stems, processed into powder, and macerated with ethanol 80%. After that, the extraction was performed in three different concentrations, which were 25%, 50%, and 100%. five groups of Wistar rats were divided into Healthy wistar rats (control negative) Wistar rats suffering from periodontitis (control positive), Wistar rats suffering from periodontitis treated with 0.1 ml of 25% brotowali extract (treatment I), suffering from periodontitis treated with 0.1 ml of 50% brotowali extract (treatment II), and Wistar rats suffering from periodontitis treated with 0.1 ml of 100% brotowali extract (treatment III). The histological slide of each sample then was made to determine the number of macrophages cells. Results: One-Way ANOVA analysis revealed a notable difference among the groups. Tukey HSD Test (Post Hoc Test) showed no significant difference between (treatment group I) and (treatment group II). Conclusion: Brotowali extract may influence macrophage cells count of Wistar rats caused by A. actinomycetemcomitans bacteria.

Effect of Acute Exposure of Cigarette Smoke on Endometrium Thickness and Number of Ovarian Follicles in Rats

Background: Cigarrete smoke has been ascociated with many health-related problems especially in long time use. One of many health problems that are affected is the reproductive system that give negative impact in fertility. However, there are limited studies about this in acute exposure of cigarette smoke. Objective: The aim of this study is to investigate the effect of acute exposure of cigarette smoke on endometrial thickness and number of ovarian follicles in rats. Methods: Thirty adult female rats were randomly divided into 3 groups: P0, P1 and P2. The P1 and P2 groups were exposed to cigarette smoke for 15 days with dose of 2 and 4 cigarettes daily, respectively. Vaginal swab was done twice, before and after the last treatment, to determine the proestrus phase of the animals. Histological slide of uterus and ovarium using Haematoxylin Eosin staining were observed under light microscop. Number of ovarian follicles were measured, and the thickness of endometrium was meassured using CellSens software version 2.3 (Olympus Life Science). Statistical analysis of data was performed using MANOVA in SPSS software version 23 for Windows. Results: There were no significant differences in both endometrial thickness and number of ovarian follicles between 3 groups (p>0.05). However, we found that there were a downward trend of some variables (endometrial thickness, number of primary follicles, de Graff follicles, and corpus luteum) from P0 to P2. Conclusion: We concluded that acute exposure of cigarette smoke not yet influenced the female reproductive organ, based on histological appearance of endemotrium and ovarium.

Experience of application of remote technologies in an educational process at the histology department in COVID–19 pandemic

Objective – to study the organization peculiarities of the educational process at the Histology Department using remote technologies among foreign 2nd-year students of the medical faculty of the HSEE of Ukraine “National Pirogov Memorial Medical University”, Vinnytsia.Material and methods. During the research, 90 foreign students (59 boys and 31 girls) aged 18.40 ± 0.17 years were interviewed. The survey was conducted by questionnaire in English, which consisted of 8 questions that highlighted the satisfaction and adaptation of foreign students to the learning process using distance technology (Microsoft Teams platform).Results. The study showed that the vast majority of respondents had no difficulties in mastering the interface of the Microsoft Teams platform. This, in turn, indicates the fast adaptive capability of foreign students to master remote technologies in a COVID-19 pandemic. In 43.33% of students, the subjective assessment of their knowledge is quite critical since they remained, to a certain extent, dissatisfied with the level of expertise that they received during distance learning. The survey showed that 62.69% of students were satisfied with the organization of distance learning at the histology department. The lack of full-fledged practical skills (working with a real microscope and self-recognition of all elements of the histological slide) creates a "practical deficiency" for the student.Conclusion. During the COVID-19 pandemic, distance learning in higher medical education institutions is a necessary element of the continuous education process, but it needs to be improved and cannot be a full-fledged replacement for its traditional forms.

Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia

Background: Using immunohistochemical stains to target specific breast cancer markers has become indispensable for evaluation of small diagnostic tissue specimens, and therefore novel marker cocktails for specific breast cancers are required. This study was conducted to assess the immunoexpression of P63 and SOX2 in triple negative breast cancer (TNBC), and to evaluate the predictive diagnostic value of these markers for specific types of TNBC.Methods: Histological slides and paraffin blocks of TNBC cases were collected from Dr. Hasan Sadikin Hospital, Bandung, Indonesia from 5-years period (2011-2015). Each histological slide was subjected to immunohistochemical staining for P63 (nucleus and cytoplasm) and SOX2 (nucleus), with specific primer antibodies. Immunoexpression of P63 and SOX2 was evaluated using immunoreactivity scoring. Associations between P63 and SOX2 immunoexpression and TNBC types were assessed using Mann Whitney tests. In addition, the predictive diagnostic values of these markers were assessed.Results: Forty TNBC histological slides were included, and 23 (57.5%) were Basal-like type TNBC and 17 (42.5%) were Non basal-like type TNBC. Immunoexpression of P63 nucleus and SOX2 was not different between types of TNBC. However, immunoexpression of P63 in the cytoplasm in Basal-like type TNBC was significantly higher than in Non basal-like type TNBC (p=0.021). Predictor diagnostic value analysis suggested that immunoexpression of P63 in cytoplasm had 56.5% sensitivity and 70.6% specificity for diagnosing Basal-like type TNBC, with area under curve of 0.64. Conclusions: Immunoexpression of P63 in the cytoplasm has a relatively weak diagnostic value to discriminate Basal-like and Non basal-like types of TNBC.

Effect of Laserpuncture Shoot on Reproduction Point of Male Mojosari Duck (Anas Platyrhynchos) on The Numbers of Spermatogonium Cells and Seminiferous Tubules Diameter

The Objective of this research was to know the effect of laserpuncture on reproduction point of male Mojosari duck on the number of spermatogonium cells and seminiferous tubule diameter of the testes. This research using completely randomized design with eighteen male Mojosari ducks which divided into three group randomly. Group I (P0) without any treatment of laserpuncture, Group II (P1) with 0.2 J dose of laserpuncture, and Group III (P2) with 0.4 J dose of laserpuncture. Laserpuncture shooting was done on reproduction point with three days interval in one month. On the thirty-first day, the duck was dissected so that the testes can be taken then made by the histological slide of testes. The obtained data processed by Analysis of Variant (ANOVA) followed with Duncan. Research result showed that laserpuncture shoot on the reproduction point of five-month-old male Mojosari duck decreases the number of spermatogonium cells and seminiferous tubule diameter of the testes. Keyword: laserpuncture, Mojosari duck, spermatogonium, seminiferous tubule

Effect of L-Arginine on the Thickness Iliac Arteries Wall Post Fogarty Balloon Embolectomy Catheter in Rabbit (Oryctolagus cuniculus)

The effect of the chronic administration of L-arginine on iliac arteries thickness after surgical embolectomy using Fogarty balloon catheter were examined. Sixteen New Zealand White male rabbits underwent unilateral iliac artery surgical embolectomy using Fogarty balloon catheter. Then the animals were divided into two group of control (P0) fed with standard rabbit diet and treatment (P1) standard rabbit diet added with L-arginine in their drinking water 4 for weeks. The iliac arteries were harvested for histological slide preparation with routine method, observed under the microscope attached with Nikon camera and measured using NIS-Element BR application in 8 zones. Data were collected and analyzed by independent T-test. The result of the vessel thickness was significantly decreased on the P1 (234.1± 18.1 µm) than P0 (411.6 ± 191.7 µm). Furthermore, the thickness of intimal was significantly decrease in P1 (140.9± 27.0 µm) compared with P0 (304.3± 215.7 µm). As a result, there was no significant different on the thickness of medial between P1 (93.2± 15.7 µm) and P0 (107.3± 101.5 µm). The results demonstrate that L-arginine decreased iliac artery wall thickness after surgical embolectomy with Fogarty balloon catheter in rabbit. Keywords: L-arginine ; nitric oxide ; Fogarty balloon catheter ; vascular smooth muscle cell proliferation.

Immunoexpression of P63 and SOX2 in triple-negative breast cancers, Indonesia

Background: Using immunohistochemical stains to target specific breast cancer markers has become indispensable for evaluation of small diagnostic tissue specimens, and therefore novel marker cocktails for specific breast cancers are required. This study was conducted to assess the immunoexpression of P63 and SOX2 in triple negative breast cancer (TNBC), and to evaluate the predictive diagnostic value of these markers for specific types of TNBC.Methods: Histological slides and paraffin blocks of TNBC cases were collected from Dr. Hasan Sadikin Hospital, Bandung, Indonesia from 5-years period (2011-2015). Each histological slide was subjected to immunohistochemical staining for P63 (nucleus and cytoplasm) and SOX2 (nucleus), with specific primer antibodies. Immunoexpression of P63 and SOX2 was evaluated using immunoreactivity scoring. Associations between P63 and SOX2 immunoexpression and TNBC types were assessed using Mann Whitney tests. In addition, the predictive diagnostic values of these markers were assessed.Results: Forty TNBC histological slides were included, and 23 (57.5%) were Basal-like type TNBC and 17 (42.5%) were Non basal-like type TNBC. Immunoexpression of P63 nucleus and SOX2 was not different between types of TNBC. However, immunoexpression of P63 in the cytoplasm in Basal-like type TNBC was significantly higher than in Non basal-like type TNBC (p=0.021). Predictor diagnostic value analysis suggested that immunoexpression of P63 in cytoplasm had 56.5% sensitivity and 70.6% specificity for diagnosing Basal-like type TNBC, with area under curve of 0.64. Conclusions: Immunoexpression of P63 in the cytoplasm has a relatively weak diagnostic value to discriminate Basal-like and Non basal-like types of TNBC.