MicroRNA-367 Inhibits Breast Cancer and Promotes Apoptosis by Targeting AT-Rich Interactive Domain-Containing Protein 1B

Introduction: Breast cancer (BC) developed in the glandular epithelial tissue of breast. microRNA (miR)-367 is an important player in cancer progression, but has never been studied in BC. This experiment tries to probe the mechanism of miR-367 in BC treatment with downstream target gene. Materials and Methods: Human BC cell lines and healthy breast epithelium cells were applied in this study. After the transfection of miR-367 inhibitor or mimic into BC cells, functional assays were conducted to measure cell growth. Afterwards, flow cytometry was employed in apoptosis verification. Then, target relation between miR-367 and ARID1B was certified. Furthermore, ARID1B level was also measured. Results: miR-367 was underexpressed in human BC cells (p < 0.05). Besides, overexpressed miR-367 inhibited BC cell proliferation and encouraged apoptosis, while underexpressed miR-367 led to an opposite outcome (p < 0.05). This experiment then implied that miR-367 dramatically suppressed the activity of cell transfected with ARID1B-wild type. miR-367 overexpression quenched ARID1B level in BC cells; while silencing miR-367 upregulated ARID1B expression (p < 0.05). Conclusion: Our experiment discovered that miR-367 quenched BC cell growth and promoted apoptosis by targeting ARID1B. This investigation may provide novel insights in BC treatment.

Human genetic variants associated with COVID-19 severity are enriched in immune and epithelium regulatory networks

Human genetic variants can influence the severity of infection with SARS-COV-2. Several genome-wide association studies (GWAS) have been conducted to identify human risk loci that may be involved with COVID-19 severity. However, candidate genes were investigated in the genomic proximity of each locus without considering their functional cellular contexts. Here, we compiled regulatory networks of 77 human contexts to interpret these risk loci by revealing their relevant contexts and associated transcript factors (TF), regulatory elements (REs), and target genes (TGs). 21 human contexts were identified to be associated with COVID-19 severity and grouped into two categories: immune cells and epithelium cells. We further investigated the risk loci in regulatory network of immune cells, epithelium cells and their crosstalk. Two genomic clusters, chemokine receptors cluster and OAS cluster showed the strongest association with COVID-19 severity in the context specific regulatory networks.

A Single-Cell Transcriptome Atlas of the Human Retinal Pigment Epithelium

Human retinal pigment epithelium cells are arranged in a monolayer that plays an important supporting role in the retina. Although the heterogeneity of specific retinal cells has been well studied, the diversity of hRPE cells has not been reported. Here, we performed a single-cell RNA sequencing on 9,302 hRPE cells from three donors and profiled a transcriptome atlas. Our results identified two subpopulations that exhibit substantial differences in gene expression patterns and functions. One of the clusters specifically expressed ID3, a macular retinal pigment epithelium marker. The other cluster highly expressed CRYAB, a peripheral RPE marker. Our results also showed that the genes associated with oxidative stress and endoplasmic reticulum stress were more enriched in the macular RPE. The genes related to light perception, oxidative stress and lipid metabolism were more enriched in the peripheral RPE. Additionally, we provided a map of disease-related genes in the hRPE and highlighted the importance of the macular RPE and peripheral RPE clusters P4 and P6 as potential therapeutic targets for retinal diseases. Our study provides a transcriptional landscape for the human retinal pigment epithelium that is critical to understanding retinal biology and disease.

Phylogenetic Analysis And Searching Bovine Papillomaviruses In Teat Papillomatosis Cases In Cattle By Histopathological, Immunohistochemical And Transmission Electron Microscopy Methods

Papillomaviruses are epitheliotropic viruses causing proliferations in skin, mucosa and various internal organs in different animal species. Especially due to lesions it causes in teats of cattle, it leads to important economical losses in milk sector. In this study, the aim was to diagnose bovine papillomaviruses (BPVs) causing teat papillomas in cattle by immunohistochemical, transmission electron microscopy (TEM) and molecular methods and to detect the defect on tissues by the virus using histopathological method. In addition to this, sequence analysis of the isolated field strains were to be carried out and their genetic and phylogenetic closeness with the strains within the literature were to be detected. After confirming teat papillomatosis in the collected samples using histopathological and immunohistochemical methods, other diagnosis methods were then used. During the TEM examination of teat lesions, intranuclear virus particles were seen in epithelium cells. After carrying out PCR using degenerate primers and type specific primers, 7 samples were detected as positive for BPV and these samples were used for typing using sequence analysis/PCR with type-specific primers. Within these analysis, three out of seven BPV isolates we collected from infected teat tissues of different cattle were detected as BPV-6, two as BPV-10, one as BPV-2 and one as BPV-8. Five isolates we isolated during sequence analysis of positive samples were found in Xipapillomavirus 1 genus, one in Epsilonpapillomavirus 1 genus and another in Deltapapillomavirus genus. As a result, in molecular diagnosis of BPV that takes place in etiology of teat papillomas, using type specific primers proved to be useful following the usage of genotyping in molecular diagnosis of BPV and generate primers in characterization. Detecting BPV types and their prevalence, taking biosafety measures in animal breeding and giving importance to vaccine studies was considered essential.

Investigation of Differentiated Embryonic Stem Cells Growth on Optimized Porous Polymeric Bed with Fuzzy System

Introduction: Age-related macular degeneration (AMD) is one of the retina diseases in which retinal pigment epithelium cells are degraded and lead to blindness. Available treatments only slow down the progression of it. In this study, human embryonic stem cells (hESCs) differentiated into retinal pigment epithelium cells were cultured on a polycaprolactone scaffold. Methods: The optimization of the diameter of the produced scaffolds by electrospinning method was done using the fuzzy method for the first time. To improve cell adhesion and proliferation, related parameters to alkaline hydrolysis method were optimized and hydrophobic surface of scaffold was modified. After in vitro analysis, cells were cultured on different groups of scaffolds. In vivo analyses were done and cells culture on scaffolds observed. Results: The optimal parameters for the scaffold based on the fuzzy model were 18.1 kV for voltage, 0.07 g / ml for solution concentration and 115 nm for scaffold diameter, respectively. The immersion time of the scaffold in alkaline solution and concentration of solution were measured 97 min and 3.7 M, respectively. The treated scaffold had a higher degradation rate and water adsorption. MTT-Assay results showed that scaffolds with modified surfaces had a higher amount of cell viability and proliferation after 7 days. SEM image results confirmed this finding after almost two months. Additionally, the results of ICC test showed that after passing this time, cells kept their RPE and epithelium. Conclusion: Based on the results, the hydrolyzed scaffold is a suitable substrate for cell proliferation and can be a good option for AMD treatment.

Internalization and Transport of PEGylated Lipid-Based Mixed Micelles across Caco-2 Cells Mediated by Scavenger Receptor B1

The aim of this study was to get insight into the internalization and transport of PEGylat-ed mixed micelles loaded by vitamin K, as mediated by Scavenger Receptor B1 (SR-B1) that is abundantly expressed by intestinal epithelium cells as well as by differentiated Caco-2 cells. Inhibition of SR-B1 reduced endocytosis and transport of vitamin-K-loaded 0%, 30% and 50% PEGylated mixed micelles and decreased colocalization of the micelles with SR-B1. Confocal fluorescence microscopy, fluorescence-activated cell sorting (FACS) analysis, and surface plasmon resonance (SPR) were used to study the interaction between the mixed micelles of different compositions (varying vitamin K loading and PEG content) and SR-B1. Interaction of PEGylated micelles was independent of the vitamin K content, indicating that the PEG shell prevented vitamin K exposure at the surface of the micelles and binding with the receptor and that the PEG took over the micelles’ ability to bind to the receptor. Molecular docking calculations corroborated the dual binding of both vita-min K and PEG with the binding domain of SR-B1. In conclusion, the improved colloidal stability of PEGylated mixed micelles did not compromise their cellular uptake and transport due to the affinity of PEG for SR-B1. SR-B1 is able to interact with PEGylated nanoparticles and mediates their subsequent internalization and transport.

The Role of Lactic Acid on Wound Healing, Cell Growth, Cell Cycle Kinetics, and Gene Expression of Cultured Junctional Epithelium Cells in the Pathophysiology of Periodontal Disease

Lactic acid (LA) is short-chain fatty acid, such as butyric acid and propionic acid, that is produced as a metabolite of lactic acid bacteria, including periodontopathic bacteria. These short-chain fatty acids have positive effects on human health but can also have negative effects, such as the promotion of periodontal disease (PD), which is caused by periodontal pathogens present in the gingival sulcus. PD is characterized by apical migration of junctional epithelium, deepening of pockets, and alveolar bone loss. Thus, the junctional epithelial cells that form the bottom of the gingival sulcus are extremely important in investigating the pathophysiology of PD. The aim of this study was to investigate the effect of LA on wound healing, cell growth, cell cycle kinetics, and gene expression of cultured junctional epithelium cells. The results showed that stimulation with 10 mM LA slowed wound healing of the junctional epithelial cell layer and arrested the cell cycle in the G0/G1 (early cell cycle) phase, thereby inhibiting cell growth. However, cell destruction was not observed. LA also enhanced mRNA expression of integrin α5, interleukin (IL)-6, IL-8, intercellular adhesion molecule-1, and receptor activator of nuclear factor kappa-B ligand. The results of this study suggest that stimulation of junctional epithelial cells with high concentrations of LA could exacerbate PD, similarly to butyric acid and propionic acid.

Phylogenetic Analysis and Searching Bovine Papillomaviruses in Teat Papillomatosis Cases in Cattle by Histopathological, Immunohistochemical and Transmission Electron Microscopy Methods

Papillomaviruses are epitheliotropic viruses causing proliferations in skin, mucosa and various internal organs in different animal species. Especially due to lesions it causes in teats of cattle, it leads to important economical losses in milk sector. In this study, the aim was to diagnose bovine papillomaviruses (BPVs) causing teat papillomas in cattle by immunohistochemical, transmission electron microscopy (TEM) and molecular methods and to detect the defect on tissues by the virus using histopathological method. In addition to this, sequence analysis of the isolated field strains were to be carried out and their genetic and phylogenetic closeness with the strains within the literature were to be detected. After confirming teat papillomatosis in the collected samples using histopathological and immunohistochemical methods, other diagnosis methods were then used. During the TEM examination of teat lesions, intranuclear virus particles were seen in epithelium cells. After carrying out PCR using degenerate primers and type specific primers, 7 samples were detected as positive for BPV and these samples were used for typing using sequence analysis/PCR with type-specific primers. Within these analysis, three out of seven BPV isolates we collected from infected teat tissues of different cattle were detected as BPV-6, two as BPV-10, one as BPV-2 and one as BPV-8. Five isolates we isolated during sequence analysis of positive samples were found in Xipapillomavirus 1 genus, one in Epsilonpapillomavirus 1 genus and another in Deltapapillomavirus genus. As a result, in molecular diagnosis of BPV that takes place in etiology of teat papillomas, using type specific primers proved to be useful following the usage of genotyping in molecular diagnosis of BPV and generate primers in characterization. Detecting BPV types and their prevalence, taking biosafety measures in animal breeding and giving importance to vaccine studies was considered essential.