Preparation Of Tissue Sections Research Articles

Precise quantification of skeletal muscle fibers reveals the physiological basis for growth rate discrepancies in broilers1

Skeletal muscle is composed of multinucleated muscle fibers, which play a crucial role in determining the quality of meat products in livestock. Quantifying the total number of muscle fibers (TNM) is essential for understanding muscle composition, yet remains challenging in poultry, particularly due to the size of the livestock that complicates the preparation of tissue sections for analysis and renders the counting process laborious. Our previous study developed an automatic muscle fiber quantification tool powered by deep learning, named MyoV, which has addressed this bottleneck. This study aimed to employ the tool for the accurate quantification of the TNM in the pectoral muscles of slow-growing (SL), medium-growing (ML), and fast-growing (FL) broilers. Results showed that FL exhibited higher growth performance compared to ML and SL from embryonic to rearing stages. Processing of whole slide images of pectoral muscle revealed significantly higher TNM in FL and ML than in SL (P < 0.01). The TNM of FL, ML and SL were 693,568.00 ± 54,169.80, 652,122.00 ± 65,822.60 and 539,778.57±40,722.94 at 7 days of age (D7), respectively. And the TNM at D35 were 663,014.93±58,801.11, 645,784.76±80,204.34 and 507,280.29±98,092.16 of FL, ML and SL. Differences in cross-sectional area (CSA) of muscle fibers among the three groups were consistent with TNM results. Correlation analysis showed a correlation coefficient of 0.73-0.89 between body weight (BW) and TNM and a correlation coefficient of 0.78-0.87 between BW and CSA. These findings directly indicate that the number of muscle fibers in broilers is an important foundation for their rapid growth and development. This study precisely quantifies the muscle fiber number of important skeletal muscle in poultry for the first time, providing the direct evidence for the physiological basis of rapid development in broilers and offering important data support for further in-depth researches on muscle fiber development.

Pathological and phylogenetic characterization of a rare fowl adenovirus (FAdV-8b) associated with inclusion body hepatitis in naturally infected Meleagris gallopavo.

Adenoviruses are a diverse group of viruses that can cause a variety of diseases in poultry, including respiratory and gastrointestinal infections. In turkeys (Meleagris gallopavo), adenoviruses commonly cause hemorrhagic enteritis and, rarely, inclusion body hepatitis. In this study, we investigated fowl adenoviruses (FAdVs) circulating in turkeys in Egypt. Following clinical examination of 500 birds, a portion of the hexon gene was amplified from four out of 50 samples from diseased birds (8%), and one amplicon that produced a strong band was selected for sequencing. Molecular and phylogenetic analysis revealed that the virus in that sample belonged to serotype FAdV-8b. Histopathological and immunohistochemical examinations of prepared tissue sections were performed to confirm the pathological findings. Diseased birds exhibited ruffled feathers, low body weight, a crouching posture, and diarrhea. Gross examination revealed petechial hemorrhage on the spleen, swollen pale liver, and congested intestine. Microscopic examination revealed the presence of eosinophilic and basophilic intranuclear inclusion bodies, nuclear pyknosis, and apoptotic bodies in the liver, congestion, hemorrhage, and fibrosis in the lungs, and desquamation of enterocytes. The presence of viral antigens in the liver, lungs, and intestine was confirmed by immunohistochemistry. To our knowledge, this is the first report of the characterization of an outbreak of inclusion body hepatitis in turkeys (hybrid converter breeds) due to FAdV-8b in Egypt. This finding raises an epidemiological alarm, necessitating further studies, including full-genome sequencing, to trace the virus's origin and genetic diversity.

Expression of PD-L1 (DAKO 28-8) in Urothelial Carcinoma of Urinary Bladder

Introduction: Around 90% of bladder cancers are urothelial carcinoma. Recent advancements in treatment have shifted towards targeted therapies, particularly immunotherapies targeting PD-L1 (Programmed Cell Death Ligand 1). PD-L1, a ligand for PD-1 receptors, helps cancer cells evade the immune system. Various factors, including genetic alterations, lead to its upregulation in cancer cells. Immunotherapies aim to block this pathway, making cancer cells more susceptible to the immune response. Objective: The study was designed to assess the frequency of PD-L1 expression by immunohistochemistry in urothelial carcinoma of urinary bladder and its association with histological grades. Methodology: This study was conducted at Department of Pathology, Pakistan Institute of Medical Sciences Islamabad. Fifty-six cases of urothelial carcinoma were included. Immunohistochemistry using PD-L1 Dako 28-8 clone was applied to prepared tissue sections. PD-L1 positivity was determined by brown membrane staining in tumor cells, scored using the Tumor Proportion Score (TPS). Results: The median patient age was 63±7 years, with the highest frequency (58.9%) of PD-L1 expression occurring in the age range of 56 to 65 years. However, there was no significant association between PD-L1 expression and age groups. Out of 56 cases, 64.3% had a TPS (Tumor Proportion Score) of ≥5%. Among the 76.7% high-grade cases, 72% had a TPS ≥5%, whereas 28% had a TPS &lt;5%. Among the 23.2% low-grade cases, 61.5% had a TPS &lt;5%, and 38.5% had a TPS ≥5%. Conclusion: Sixty-four percent urothelial carcinoma cases showed PD-L1 expression. Significant association (p value = 0.02) between PD-L1 expression and high-grade urothelial carcinoma was found using Pearson’s Chi-square test.

An approach for improvement of the accuracy of cancer gene panel testing.

Tissue-based comprehensive genomic profiling (CGP) is increasingly being employed for genotype-directed therapies in patients with advanced cancer. However, tissue availability may limit their potential applications. In Japan, the cost of cancer gene panel tests is covered by public insurance for patients diagnosed with advanced solid tumors once in their lifetime. Therefore, it is essential to improve the success rate (reportability) and accuracy of CGP tests. The purpose of this study was to identify the factors associated with efficient and accurate CGP testing using relevant information obtained from real-world data. This study included 159 samples analyzed using tumor-only panel FoundationOne® CDx cancer genome profiling (F1CDx) and 85 samples analyzed using matched-pair panel OncoGuide™ NCC Oncopanel system (NCCOP) at St. Marianna University Hospital. Sample characteristics (fixation conditions, storage period, histology, tumor cell ratio, and genomic tumor cell content), CGP performance, and quality control status were evaluated across all 244 tested samples. In 237/244 samples (97.1%), CGP testing results were successfully obtained [F1CDx, 99.4% (158/159) and NCCOP, 92.9% (79/85)]. An increased number of fibroblasts, inflammatory cells, and necrotic tumor cells, long-term storage, and/or prolonged fixation of tissue sections were involved in the unreported results and/or qualified CGP results. In addition, a negative correlation between median insert size values and ΔΔCq was observed in the NCCOP system. We identified various factors associated with efficient and accurate CGP testing using relevant information obtained from real-world data, suggesting that thorough selection and preparation of tissue sections could optimize CGP and maximize useful information.

Biomedical laboratory scientists and technicians in digital pathology - Is there a need for professional development?

Digital pathology (DP) is moving into Danish pathology departments at high pace. Conventionally, biomedical laboratory scientists (BLS) and technicians have prepared tissue sections for light microscopy, but workflow alterations are required for the new digital era with whole slide imaging (WSI); digitally assisted image analysis (DAIA) and artificial intelligence (AI). We aim to explore the role of BLS in DP and assess a potential need for professional development. We investigated the roles of BLS in the new digital era through qualitative interviews at Danish Pathology Departments in 2019/2020 before DP implementation (supported by a questionnaire); and in 2022 after DP implementation. Additionally, senior lecturers from three Danish University Colleges reported on how DP was integrated into the 2023 bachelor's degree educational curricula for BLS students. At some Danish pathology departments, BLS were involved in the implementation process of DP and their greatest concerns were lack of physical laboratory requirements (69%) and implementation strategies (63%). BLS were generally positive towards working with DP, however, some expressed concern about extended working hours for scanning. Work-task transfers from pathologists were generally greeted positively from both management and pathologists; however, at follow-up interviews after DP implementation, job transfers had not been effectuated. At Danish university colleges, DP had been integrated systematically in the curricula for BLS students, especially WSI. Involving BLS in DP implementation and development may benefit the process, as BLS have a hands-on workflow perspective with a focus on quality assurance. Several new work opportunities for BLS may occur with DP including WSI, DAIA and AI, and therefore new qualifications are warranted, which must be considered in future undergraduate programmes for BLS students or postgraduate programmes for BLS.

GNTD: reconstructing spatial transcriptomes with graph-guided neural tensor decomposition informed by spatial and functional relations

Spatially-resolved RNA profiling has now been widely used to understand cells’ structural organizations and functional roles in tissues, yet it is challenging to reconstruct the whole spatial transcriptomes due to various inherent technical limitations in tissue section preparation and RNA capture and fixation in the application of the spatial RNA profiling technologies. Here, we introduce a graph-guided neural tensor decomposition (GNTD) model for reconstructing whole spatial transcriptomes in tissues. GNTD employs a hierarchical tensor structure and formulation to explicitly model the high-order spatial gene expression data with a hierarchical nonlinear decomposition in a three-layer neural network, enhanced by spatial relations among the capture spots and gene functional relations for accurate reconstruction from highly sparse spatial profiling data. Extensive experiments on 22 Visium spatial transcriptomics datasets and 3 high-resolution Stereo-seq datasets as well as simulation data demonstrate that GNTD consistently improves the imputation accuracy in cross-validations driven by nonlinear tensor decomposition and incorporation of spatial and functional information, and confirm that the imputed spatial transcriptomes provide a more complete gene expression landscape for downstream analyses of cell/spot clustering for tissue segmentation, and spatial gene expression clustering and visualizations.

Effect of vitamin C on histomorphometric changes of testis, epididymis, prostate, and seminal vesicle induced by acrylamide in rat.

Acrylamide (ACR) has adverse effects on the rat testis. This study aimed to assess the impact of ACR and vit C exposure on reproductive organs in rats. In this experimental study, 32 adult male rats were used. The animals were divided into 4 groups (n = 8): (1) control group, (2) ACR (10mg/kg) group, (3) vit C (200mg/kg), (4) ACR (10mg/kg) + vit C (200mg/kg) daily for 5weeks by gavage. After the administration period, testis, prostate, seminal vesicle, and epididymis of animals are removed; after preparing tissue sections, the structural changes of the tissues are examined by stereology. Data were analyzed using one-way ANOVA followed by the Tukey test in SPSS software. A value of p ≤ 0.05 was considered significant. The testis weight, volume (mm3), and the mean Johnsen score showed a significant decrease in comparison with the control group and vit C-treated group. These parameters were increased in ACR + vit C group. The number of spermatogonia, spermatocyte, spermatid, and Sertoli and Leydig cells in ACR-treated group showed a significant decrease in comparison with the control and vit C-treated groups. The number of these cells was increased in the ACR + vit C group. Epithelium height and folding of prostate and seminal vesicle in the ACR-treated group were decreased. Epithelium lost its integrity. In the ACR + vit C group, histopathological changes were decreased. Seminal vesicle of ACR + vit C-treated group showed mild degeneration and rupture in epithelium integrity. The epididymis of ACR + vit C group also showed mild degeneration and rupture in epithelium integrity.

Comparative evaluation of the effect of three types of fixatives (formalin, Bouin and Carnoy) on histomorphological features of various viscera.

Tissue shrinkage is one of the problems in preparing tissue sections. This study compares the use of 10% formalin, Bouin and Carnoy as fixatives on several mouse tissues to determine histomorphological features. In this experimental study, liver, kidney, heart, lung, testicle, spleen, brain and cartilage tissues were isolated from five BALB/c mice. Then, they were fixed with three types of fixatives. After dehydrating, clarifying and embedding, all samples were stained with haematoxylin and eosin. Then, the tissue structure of the viscera was evaluated qualitatively. The results showed that each fixative is more suitable for evaluating a specific part of the tissue. However, relative shrinkage appeared in the tissue sections fixed with 10% Formalin, (1) in the heart as spaces between muscle fibre bundles, (2) in the liver as the dilation of the liver sinusoidal spaces, (3) in the kidney tissue as the expansion of the lumens of the convoluted proximal and distal tubules, (4) in the spleen as open spaces inside the red and white pulps and (5) in the brain as an increase in the space between the cells of the granular and pyramidal cell layers of the cortex. In tissues that were soft and fragile, such as testis, liver and brain, Bouin's fixative was more suitable. Carnoy's fixative was more suitable for the spleen and kidney tissue. Based on the study results, formalin and Bouin were more suitable for heart and cartilage tissue. Considering that in the histopathological evaluation both the cytoplasm and the nucleus are evaluated, it is suggested to choose the fixative suitable for the type of tissue.

Anti-inflammatory and collagenation effects of zinc oxide-based nanocomposites biosynthesised with Mentha longifolia leaf extract.

The integration of nanomaterials and herbal medicine has led to the design of new nanocomposites, which are therapeutically more effective. The purpose of this study was to prepare different zinc oxide (ZnO)-based nanoparticles (NPs) via Mentha longifolia extract based on gauze linen fibre and study its effects on wound healing. The textural properties, morphology, thermal stability, purity, spectroscopic and phase structure of nanoparticles were investigated. Subsequently, male Wistar rats were subjected to wounds in six different treatment groups: Group I: control; group II: ZnO/W prepared in water (W); group III: ZnO/M synthesised with Mentha longifolia (M) extract; group IV: ZnO/copper(II) oxide (CuO)/M nanocomposite synthesised with M extract; group IV: treated with ZnO/silver (Ag)/M nanocomposite; group V: treated with ZnO/Ag/M nanocomposite; and finally, group VI: treated with ZnO/CuO/Ag/M nanocomposite. In all groups, the wounds were treated for 21 days with prepared samples. Every seven days, after measuring the decreasing rate of the wound size, tissue samples from each group were taken for histopathological analysis. The prepared tissue sections were assessed by haematoxylin and eosin staining for the formation of the epidermis, dermis and muscular tissue, and Masson's Trichrome staining for the formation of collagen fibres. The results showed that the ZnO/CuO/Ag/M nanocomposite was a significantly more effective wound healing material in comparison with other samples (p<0.05). In this study, the integration of ZnO/CuO/Ag nanocomposites with secondary metabolites of Mentha longifolia gave rise to a superior combination, which could support different phases of wound healing via the regulation of cytokines and growth factors in the course of healing.

Stratifying Ferroptosis Sensitivity in Cells and Tissues with PALP.

Ferroptosis is emerging as a promising strategy for suppressing multiple types of human cancers. Rapid and accurate assessment of the relative sensitivity to ferroptosis in biological samples will accelerate the development of ferroptosis-targeted therapies. We previously demonstrated that photochemical activation of membrane lipid peroxidation (PALP) that uses high-power lasers to induce localized polyunsaturated fatty acyl (PUFA)-lipid peroxidation can efficiently report ferroptosis sensitivity in live cells and tissues in situ. Here, we describe the experimental details for PALP analysis, including preparation of tissue sections, preparation of fluorescent lipid peroxidation reporter, sample staining, lipid peroxidation induced by laser source, and data processing. We envision predicting the relative sensitivity to ferroptosis of cellular and tissue samples is potentially useful for basic research and clinical investigations.

Severe deterioration in sperm parameters and testes of rats administered naproxen and diclofenac at pre-puberty: An experimental study.

Although naproxen and diclofenac are the most commonly used nonsteroidal drugs, their toxicity affecting male reproductivity has not been sufficiently studied, particularly when used in pre-puberty. This study aims to investigate the toxic effects of naproxen and diclofenac on sperm parameters and testes in pre-pubertal rats. In this experimental study, 15 pre-pubertal male albino rats (aged 5 wk, weighted 70-80 gr) were used. The animals were divided in to 3 groups (n = 5/each) of control (0.1 ml dimethyl sulfoxide), naproxen (50 mg/kg), and diclofenac sodium (5 mg/kg), and were orally administered with these drugs every day for 3 wk. Epididymal spermatozoa were taken to assess sperm count, viability, and morphology. After preparing tissue sections, testicular histopathological and histomorphometric analyses were performed. The body weights of rats in both naproxen and diclofenac groups significantly decreased (p 0.001 and p = 0.03, respectively) in comparison to the control group. Remarkably, the testis weight and total sperm number significantly decreased (p = 0.002, and p = 0.004 respectively) in the naproxen-administered rats only. The sperm viability percentage decreased significantly in both diclofenac and naproxen-administered groups (p 0.001 and p = 0.03 respectively). Moreover, there was a significant increase (p 0.001) in the percentage of sperm morphological anomalies in both drug-administered groups. Also, the histological and morphometric findings exhibited severe histopathological appearances in the testes and seminiferous tubules parameters in both drug-administered groups.>. Naproxen and diclofenac administrations of rats before their puberty induce considerable harm to sperm parameters and testicular histology and morphometry. These severe toxic effects can lead to potential infertility.

Abstract 232: Qualitative Single-cell Assessment By Imaging Mass Cytometry (imc) Analysis Reveals Senescence-associated Stemness (sas) Induced By Erk5 S496 Phosphorylation In Atherosclerotic Plaque

The central role of ERK5 S496 phosphorylation in promoting senescence has been reported. Recently the conventional model of senescence as an event of cell cycle arrest has been challenged, given that senescence induced by various stresses reprograms cancer cells to acquire senescence-associated stemness (SAS), which allows them to escape senescence-induced cell cycle arrest with enhanced growth potential. To determine the role of ERK5 S496 phosphorylation in SAS, we utilized imaging mass cytometry (IMC) by using antibodies against various cell surface markers, senescence, and proliferation. We prepared tissue sections from the plaques in wild-type (WT) and ERK5 S496A knock-in (KI) mice, and the ablated plaque materials were evaluated by the CyTOF mass cytometer. We used the VISIOPHARM program, and single-cell features were computationally segmented using a watershed algorithm. Nine clusters were detected in WT (total 7903 cells) and ERK5 S496A KI (total 6166 cells) plaques. Single-cell analysis of p53 and Ki67 revealed three groups of the cells with different pattern of p53 and Ki67 expression (Fig.A). We calculated log scaled ratio of Ki67 and p53 of selected cells by log 10 (expression of Ki67/ expression of p53), and further calculated one-dimension density of this log scaled ratio to cluster cells into three groups by using cutoff of ration with log 10 (-1.01) and log 10 (-0.024) (Fig.B). Group 2 myeloid cells showed that p53 and Ki67 were co-expressed lineally, suggesting that this group of cells escaped from the growth-suppression effects of p53, which is also known as a senescence marker. Of note, the % of ERK5 S496A KI cells in group 2 were significantly lower than those in WT (Fig.C). Also, in vitro oxidized LDL remarkably increase β-gal + Ki67 + cells, which was significantly inhibited in ERK5 S496A KI macrophage compared to those in WT. Collectively, these data implicates ERK5 S496 phosphorylation in a unique role-regulation of SAS during atherogenesis.

Identification of the function of γδ1 T cells in the lung cancer microenvironments.

To investigate whether γδ1 T cells derived from lung cancer tissues have immunosuppressive function and to verify the mechanism of immunosuppressive effect. Fresh lung cancer tissue samples were collected, some of them were prepared tissue sections, the others were isolated and amplified into TILs cells, γδ1 T cells were isolated from TILs cells by immunomagnetic beads kits, and then cloned and amplified. The immunomodulatory effects of γδ1 T cells on naive and effector CD4+ T cells were detected by immunohistochemistry, flow cytometry, CCK8, ELISA and transwell culture. A high proportion of γδ1 T cells was found in lung cancer tissues. The cultural supernatants of γδ1 T cells could inhibit the proliferation of naive CD4+ T cells and decrease the secretion level of IL-2 by effector CD4+ T cells. Further studies showed that the expression levels of IL-8, MIP-1α, MIP-1β and RANTES were higher than that of IFN-γ, GM-CSF and TNF-α, TNF-β, however, their neutralizing antibodies could not block the immunosuppressive activity of the supernatant. γδ1 T cells play an negative immunoregulation function in lung cancer microenvironments, and have obvious immunosuppressive effects on proliferation and cytokine release of naive CD4+ T cells and effector CD4+ T cells. Preliminary evidence from this study suggests that the mechanism of immunosuppressive effects is mediated by the soluble factors in γδ1 T cell culture supernatants, but its exact molecular mechanism needs to be further explored.

Imputation of spatially-resolved transcriptomes by graph-regularized tensor completion.

High-throughput spatial-transcriptomics RNA sequencing (sptRNA-seq) based on in-situ capturing technologies has recently been developed to spatially resolve transcriptome-wide mRNA expressions mapped to the captured locations in a tissue sample. Due to the low RNA capture efficiency by in-situ capturing and the complication of tissue section preparation, sptRNA-seq data often only provides an incomplete profiling of the gene expressions over the spatial regions of the tissue. In this paper, we introduce a graph-regularized tensor completion model for imputing the missing mRNA expressions in sptRNA-seq data, namely FIST, Fast Imputation of Spatially-resolved transcriptomes by graph-regularized Tensor completion. We first model sptRNA-seq data as a 3-way sparse tensor in genes (p-mode) and the (x, y) spatial coordinates (x-mode and y-mode) of the observed gene expressions, and then consider the imputation of the unobserved entries or fibers as a tensor completion problem in Canonical Polyadic Decomposition (CPD) form. To improve the imputation of highly sparse sptRNA-seq data, we also introduce a protein-protein interaction network to add prior knowledge of gene functions, and a spatial graph to capture the the spatial relations among the capture spots. The tensor completion model is then regularized by a Cartesian product graph of protein-protein interaction network and the spatial graph to capture the high-order relations in the tensor. In the experiments, FIST was tested on ten 10x Genomics Visium spatial transcriptomic datasets of different tissue sections with cross-validation among the known entries in the imputation. FIST significantly outperformed the state-of-the-art methods for single-cell RNAseq data imputation. We also demonstrate that both the spatial graph and PPI network play an important role in improving the imputation. In a case study, we further analyzed the gene clusters obtained from the imputed gene expressions to show that the imputations by FIST indeed capture the spatial characteristics in the gene expressions and reveal functions that are highly relevant to three different kinds of tissues in mouse kidney.

Biologically‐synthesised ZnO/CuO/Ag nanocomposite using propolis extract and coated on the gauze for wound healing applications

Wound healing has long been recognised as a major clinical challenge for which stablishing more effective wound therapies is necessary. The generation of metallic nanocomposites using biological compounds is emerging as a new promising strategy for this purpose. In this study, four metallic nanoparticles (NPs) with propolis extract (Ext) and one without propolis including ZnO/Ext, ZnO/Ag/Ext, ZnO/CuO/Ext, ZnO/Ag/CuO/Ext and ZnO/W were prepared by microwave method and assessed for their wound healing activity on excision experimental model of wounds in rats. The developed nanocomposites have been characterised by physico-chemical methods such as X-ray diffraction, scanning electron microscopy, diffuse reflectance UV-vis spectroscopy, Fourier transform infrared spectroscopy, thermogravimetric analysis and Brunauer-Emmett-Teller analyses. The wounded animals treated with the NPs/Ext in five groups for 18 days. Every 6 days, for measuring wound closure rate, three samples of each group were examined for histopathological analysis. The prepared tissue sections were investigated by haematoxylin and Eosin stainings for the formation of epidermis, dermis and muscular and Masson's trichrome staining for the formation of collagen fibres. These findings toughly support the probability of using this new ZnO/Ag/Ext materials dressing for a wound care performance with significant effect compared to other NPs.

Evaluation of innervation in cleft affected oral mucosa

Orofacial clefts are one of the most common pathologies present at birth. The aim of the study was to evaluate the presence of PGP 9.5, substance P, VIP, CGRP, myelin and NFG in cleft affected mucosa of alveolar process, soft and hard palate, vomer and lip.&#x0D; Methods. Orofacial cleft affected mucosa tissue samples were obtained during surgical cleft correction from 21 children aged from 2 months to 9 years and 10 months. Prepared tissue sections were stained by immunohistochemistry for PGP 9.5, substance P, VIP, CGRP, myelin and NGF. The intensity of staining was graded semiquantitatively.&#x0D; Results. Mostly moderate number and numerous PGP 9.5, substance P, VIP, CGRP, myelin and NGF-containing nerve fibres and nerve fibre bundles and CGRP and NGF positive keratinocytes were detected. The most immunoreactive for PGP 9.5, myelin and NGF were nerve fibres, nerve fibre bundles and keratinocytes in lip mucosa tissue samples. Typically, PGP 9.5-containing nerve fibres were detected in subepithelium close to basal lamina, near blood vessels; thin PGP 9.5 positive nerve fibres were also observed between keratinocytes. Immunoreactivity of CGRP was higher than of substance P in nerve fibres and nerve fibre bundles. Statistically significant positive correlation was observed between all markers in subepithelium of all tissue samples.&#x0D; Conclusions. The proved statistically significant strong inter-correlation and the number of general neuropeptide-containing innervation (PGP 9.5), sensory nerves (CGRP and substance P), and parasympathetic nerves (VIP) is similar in orofacial cleft affected oral mucosa to such described in scientific literature healthy oral mucosa, except the alveolar process covering mucosa where the above innervation varies.&#x0D; Very strong and strong statistically significant positive correlations between the number of PGP 9.5, substance P, CGRP and VIP positive structures on the one hand, and NGF and myelin, on the other hand, indicate a connection between quality markers and common neuropeptide-containing, sensory and parasympathetic innervation in cleft affected oral tissue.&#x0D; The number of PGP 9.5, NGF and myelin immunoreactive neuronal structures in lip mucosa is higher than in the alveolar process, soft and hard palate and vomer mucosa, suggesting the significance of qualitative common innervation in this organ even in the cleft affected case.

Pathological Findings of Postmortem Biopsies From Lung, Heart, and Liver of 7 Deceased COVID-19 Patients.

Background. A novel coronavirus named severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been affecting almost all nations around the world. Most infected patients who have been admitted to intensive care units show SARS signs. In this study, we aimed to achieve a better understanding of pathological alterations that take place during the novel coronavirus infection in most presumed affected organs. Methods. We performed postmortem core needle biopsies from lung, heart, and liver on 7 deceased patients who had died of coronavirus disease 2019. Prepared tissue sections were observed by 2 expert pathologists. Results. Diffuse alveolar damage was the main pathologic finding in the lung tissue samples. Patients with hospitalization durations of more than 10 days showed evidence of organization. Multinucleated cells in alveolar spaces and alveolar walls, atypical enlarged cells, accumulation of macrophages in alveolar spaces, and congestion of vascular channels were the other histopathologic alteration of the lung. None of our heart biopsy samples met the criteria for myocarditis. Liver biopsies showed congestion, micro- and macro-vesicular changes, and minimal to mild portal inflammation, in the majority of cases. Conclusions. Similar to the previous coronavirus infection in 2003, the main pathologic finding in the lung was diffuse alveolar damage with a pattern of organization in prolonged cases. The SARS-CoV-2 infection does not cause myocarditis, and the ischemia of myocardium is the most probable justification of the observed pathologic changes in the heart. Liver tissue sections mostly showed nonspecific findings; however, ischemia of the liver can be identified in some cases.

Histological study of intestinal goblet cells, IgA, and CD3+ lymphocyte distribution in Huang-huai white goat.

Ten healthy adult Huang-huai white goats were selected and sacrificed by jugular vein bleeding after anaesthesia to observe the distribution characteristics of the histological structure of the intestinal mucosa, goblet cells, IgA, and CD3+ lymphocytes. Three sections of the duodenum, the jejunum, and the ileum were immediately collected and fixed with 4% paraformaldehyde for 72 h to prepare tissue sections. After haematoxylin and eosin, periodic acid Schiff, and immunohistochemical staining was performed, the distribution characteristics of goblet cells, IgA-positive cells, and CD3+ lymphocytes were observed. Results showed high columnar epithelial cells in the duodenum and jejunum of Huang-huai white goat and low columnar epithelial cells in the ileum mucosa. Mucopolysaccharides secreted by intestinal goblet cells were mainly neutral, and the number of ileum goblet cells was significantly higher than that of the duodenum and the jejunum (p < 0.05). IgA-positive cells were distributed in the lamina propria of the duodenum, and the number of cells was significantly higher than that in the jejunum and the ileum (p < 0.01). The significant difference was found between the jejunum and the ileum (p < 0.01). The CD3+ cells in the intestinal mucosa were distributed in the lamina propria mucosae, and some of the positive cells in the jejunum were distributed between epithelial cells. CD3+ cells had the largest number in the jejunal lamina propria but had the lowest number in the ileum. The jejunum was significantly higher than the duodenum (p < 0.05), and the ileum was much less than the jejunum (p < 0.01).

Immunohistochemical Expression of PTCH1 and Laminin in Oral Hyperplastic, Premalignant, Squamous Cell Carcinoma and Recurrence Lesions Samples

Objective: Laminin is a basal membrane glycoprotein that showed progressive loss of continuity from dysplasia to invasive carcinoma. The membranous receptor Patched (PTCH1) promotes the nuclear translocation and activation of the Gli family proteins. The dysregulation of hedgehog signaling reported in various cancers. This study aims to assess the immunohistochemical expression of PTCH1 and laminin in different groups of lesions of oral hyperplastic, premalignant, oral squamous cell carcinoma (OSCC) and recurrence cases. &#x0D; Methods: This study involved 35 paraffin blocks of 4 oral hyperplastic, 11 premalignant, 15 OSCC and five recurrences OSCC cases collected from Sulaimani Histopathological Centers. Prepared tissue sections were stained immunohistochemically for both PTCH1 and laminin antibodies and scored. Chi-square correlations used and the p 0.05 considered as statistically significant. &#x0D; Results: PTCH1 showed expression in all oral hyperplastic lesions. While 81.8% of oral premalignant lesions demonstrated basal and parabasal distribution with high mixed localization (72.7%), lastly 93.3% of OSCC showed positive expression and mainly found within score 2(46.6%). No significant relations detected between oral hyperplastic and premalignant lesions regarding the expression pattern, localization and intensity as p-values were 0.77, 0.09 and 0.38 respectively. Lastly, the relations between OSCC and recurrent cases to both the expression and localization parameters were non-significant, as p-values were 0.15 and 0.09, respectively. Laminin showed continuous expression at the basement membrane of the normal oral mucosa, while only (50%) of the cases revealed such expression in oral hyperplastic lesions. The oral premalignant lesions expressed 54.5% of a discontinuous pattern. The relation between the oral hyperplastic and premalignant lesions in response to laminin expression was non-significant (P=.21). A significant relation found in laminin expression between OSCC and recurrence samples (p= 0.02). Finally, a significant correlation found between PTCH1 localization and laminin expression in oral premalignant lesions (p = 0.03).&#x0D; Conclusions: The PTCH1 overexpression in all of the studied groups of lesions might give an impression of the active role of this biomarker in the progression toward malignancy. Laminin defragmentation, which started from dysplastic lesions extending to OSCC, could emphasize the role of this marker from the early precancerous stage. Furthermore, the combined PTCH1 mixed localization with discontinuous laminin expression might have a significant role in the progression of dysplastic lesions toward cancers.

Correlation Between Accumulation of Alzheimer's Disease Markers in Brain and Retina of Individuals with Late Stage of the Disease

This study was aimed to determine if there is a correlation between accumulation of various Alzheimer's disease (AD) markers in the brain and retina of individuals in the late stages of the disease. Characterizing the distribution and accumulation of these markers in the retina and brain may be important for early diagnosis of neurodegenerative diseases such as AD.We examined total tau and phosphorylated tau protein accumulations in different areas of the brain and in retinal tissues from aged individuals with AD in order to compare the intensity and sensitivity of these biomarkers.Tissues from the hippocampus and temporal lobe of four different cadavers were analyzed. Two cadavers had diagnosed Alzheimer's disease, and two cadavers served as a non‐AD control. We utilized cryosectioning to prepare tissue sections for immunostaining. Tissues obtained from individuals without a history of Alzheimer's disease or other types of dementia were used as negative controls. Detection of tau was performed using a primary antibody against human tau proteins and amyloid‐b, followed by a universal secondary antibody.Although the staining for AD markers was greater than controls for both brain tissues, the density was higher in the hippocampus than in the temporal lobe. The density of the total tau (TT) was greater than that of phosphorylated tau (PT181) in both the hippocampus and temporal lobe. We also found this to be the case in the brain tissues of a control, indicating that total tau may be a more sensitive biomarker than phosphorylated tau.These data demonstrate that total tau may be a stronger biomarker than phosphorylated tau in quantifying protein accumulations in the brains of Alzheimer's disease cadavers. This raises the intriguing possibility that there may be more sensitive biomarkers that can better predict the course of neurodegenerative diseases such as AD. Thus, future studies will concentrate on the accumulation of tau in the brain and retina of individuals during the earlier stages of Alzheimer's disease, as well as the sensitivity and accuracy of different biomarkers.Support or Funding InformationTouro University Nevada intramural grant.This abstract is from the Experimental Biology 2019 Meeting. There is no full text article associated with this abstract published in The FASEB Journal.