Ncapg dynamically coordinates the myogenesis of fetal bovine tissue by adjusting chromatin accessibility

© 2020 by the authors. Licensee MDPI, Basel, Switzerland. NCAPG is a subunit of condensin I that plays a crucial role in chromatin condensation during mitosis. NCAPG has been demonstrated to be associated with farm animal growth traits. However, its role in regulating myoblast differentiation is still unclear. We used myoblasts derived from fetal bovine tissue as an in vitro model and found that NCAPG was expressed during myogenic differentiation in the cytoplasm and nucleus. Silencing NCAPG prolonged the mitosis and impaired the differentiation due to increased myoblast apoptosis. After 1.5 days of differentiation, silencing NCAPG enhanced muscle-specific gene expression. An assay for transposase-accessible chromatinhigh throughput sequencing (ATAC-seq) revealed that silencing NCAPG altered chromatin accessibility to activating protein 1 (AP-1) and its subunits. Knocking down the expression of the AP-1 subunits fos-related antigen 2 (FOSL2) or junB proto-oncogene (JUNB) enhanced part of the muscle-specific gene expression. In conclusion, our data provide valuable evidence about NCAPG’s function in myogenesis, as well as its potential role in gene expression

bta-miR-23a Regulates the Myogenic Differentiation of Fetal Bovine Skeletal Muscle-Derived Progenitor Cells by Targeting MDFIC Gene

miR-23a, a member of the miR-23a/24-2/27a cluster, has been demonstrated to play pivotal roles in many cellular activities. However, the mechanisms of how bta-miR-23a controls the myogenic differentiation (MD) of PDGFRalpha(-) bovine progenitor cells (bPCs) remain poorly understood. In the present work, bta-miR-23a expression was increased during the MD of (PDGFRalpha-) bPCs. Moreover, bta-miR-23a overexpression significantly promoted the MD of (PDGFRalpha-) bPCs. Luciferase reporter assays showed that the 3\u27-UTR region of MDFIC (MyoD family inhibitor domain containing) could be a promising target of bta-miR-23a, which resulted in its post-transcriptional down-regulation. Additionally, the knockdown of MDFIC by siRNA facilitated the MD of (PDGFRalpha-) bPCs, while the overexpression of MDFIC inhibited the activating effect of bta-miR-23a during MD. Of note, MDFIC might function through the interaction between MyoG transcription factor and MEF2C promoter. This study reveals that bta-miR-23a can promote the MD of (PDGFRalpha-) bPCs through post-transcriptional downregulation of MDFIC

BMP4 and rosiglitazone improves adipogenesis of bovine fetal muscle derived progenitor cells

peer reviewedIntramuscular fat (IMF) content is one of the most important factors determining beef quality and price. Intramuscular adipocytes develop from mesenchymal stem cells (MSCs) in mesoderm. The mechanisms of preadipocytes differentiate into mature adipocytes to a great extent are clear, but the commitment of MSCs to preadipocytes is largely unknown. In this study, the Platelet-derived growth factor receptor α (PDGFRα) positive progenitor cells were isolated from the longissimus dorsi muscle (LM) of fetal bovine and induced adipogenesis. To optimize the in vitro IMF differentiation model, the effects of bone morphogenic protein 4 (BMP4) and rosiglitazone during differentiation were studied. Comparing with control group, progenitor cells treated with BMP4 or rosiglitazone accumulated more intracellular lipid. Furthermore, the mRNA expression level of adipocyte-specific genes also increased significantly in BMP4 or rosiglitazone treated cells. The result indicated that BMP4 and rosiglitazone could promote adipogenesis and be applied in adipogenic differentiation of fetal bovine derived progenitor cells

Robust estimation of bacterial cell count from optical density

Optical density (OD) is widely used to estimate the density of cells in liquid culture, but cannot be compared between instruments without a standardized calibration protocol and is challenging to relate to actual cell count. We address this with an interlaboratory study comparing three simple, low-cost, and highly accessible OD calibration protocols across 244 laboratories, applied to eight strains of constitutive GFP-expressing E. coli. Based on our results, we recommend calibrating OD to estimated cell count using serial dilution of silica microspheres, which produces highly precise calibration (95.5% of residuals <1.2-fold), is easily assessed for quality control, also assesses instrument effective linear range, and can be combined with fluorescence calibration to obtain units of Molecules of Equivalent Fluorescein (MEFL) per cell, allowing direct comparison and data fusion with flow cytometry measurements: in our study, fluorescence per cell measurements showed only a 1.07-fold mean difference between plate reader and flow cytometry data

Bta-miR-24-3p Controls the Myogenic Differentiation and Proliferation of Fetal, Bovine, Skeletal Muscle-Derived Progenitor Cells by Targeting

MicroRNAs modulate a variety of cellular events, including skeletal muscle development, but the molecular basis of their functions in fetal bovine skeletal muscle development is poorly understood. In this study, we report that bta-miR-24-3p promotes the myogenic differentiation of fetal bovine PDGFRα progenitor cells. The expression of bta-miR-24-3p increased during myogenic differentiation. Overexpression of bta-miR-24-3p significantly promoted myogenic differentiation, but inhibited proliferation. A dual-luciferase assay identified as a direct target of bta-miR-24-3p. Similarly, knocking down by RNA interference also significantly inhibited proliferation and promoted the differentiation of bovine PDGFRα progenitor cells. Thus, our study provides a mechanism in which bta-miR-24-3p regulates myogenesis by inhibiting expression

Bta-miR-24-3p Controls the Myogenic Differentiation and Proliferation of Fetal, Bovine, Skeletal Muscle-Derived Progenitor Cells by Targeting ACVR1B

MicroRNAs modulate a variety of cellular events, including skeletal muscle development, but the molecular basis of their functions in fetal bovine skeletal muscle development is poorly understood. In this study, we report that bta-miR-24-3p promotes the myogenic di erentiation of fetal bovine PDGFR - progenitor cells. The expression of bta-miR-24-3p increased during myogenic differentiation. Overexpression of bta-miR-24-3p significantly promoted myogenic di erentiation, but inhibited proliferation. A dual-luciferase assay identified ACVR1B as a direct target of bta-miR-24-3p. Similarly, knocking down ACVR1B by RNA interference also significantly inhibited proliferation and promoted the di erentiation of bovine PDGFR - progenitor cells. Thus, our study provides a mechanism in which bta-miR-24-3p regulates myogenesis by inhibiting ACVR1B expression

A Novel Mechanism of bta-miR-210 in Bovine Early Intramuscular Adipogenesis

Intramuscular fat (IMF) is one of the major factors determining beef quality. IMF formation is influenced by multiple conditions including genetic background, age and nutrition. In our previous investigation, bta-miR-210 was found to be increased during adipogenesis using miRNA-seq. In this study, we validated the upregulation of bta-miR-210 in platelet-derived growth factor receptor α positive (PDGFRα+) progenitor cells during adipogenic differentiation in vitro. To investigate its role in adipogenesis, bta-miR-210 mimics were introduced into progenitor cells, which resulted in enhanced intracellular lipid accumulation. Accordingly, the expression of adipocyte-specific genes significantly increased in the bta-miR-210 mimic group compared to that in the negative control group (p < 0.01). Dual-luciferase reporter assays revealed that WISP2 is a target of bta-miR-210. WISP2 knockdown enhanced adipogenesis. In conclusion, bta-miR-210 positively regulates the adipogenesis of PDGFRα+ cells derived from bovine fetal muscle by targeting WISP2

Serum organochlorine pesticide residues and risk of gallstone disease: A case-control study in Xiamen

Purpose: To investigate the association between serum organochlorine pesticide residues and risk of gallstone disease. Methods: A1:1, pair-matched, case-control study was designed. Data from 150 patients with gallstones diagnosed by abdominal ultrasonography at a single hospital from June 2009 to June 2010 were collected. A total of 150 patients without gallstones during the same period at the same hospital were recruited as the control group. Capillary gas chromatography was employed to measure the serum concentrations of dichlorodiphenyltrichloroethane (DDT) and hexachlorocyclohexane (HCH) residues. Multiple-factor conditional logistic regression analysis was conducted to estimate the relative risk of gallstones in relation to organochlorine pesticide residues in serum. Results: The percentages of p,p'-DDD and o,p'-DDT in serum of patients were significantly higher than those in serum of controls. The p,p'-DDE, alpha-HCH, and delta-HCH residues in serum of patients were also significantly increased compared with those in serum of controls. Multiple-factor conditional logistic regression analysis showed that high levels of p,p'-DDE and p,p'-DDT residues were risk factors for gallstone disease. Conclusions: A high level of organochlorine pesticide residues in serum is a potential risk factor for gallstone disease, which suggests that environmental exposure to organochlorine pesticides should be evaluated with respect to gallstone formation. (C) 2012 Elsevier Inc. All rights reserved.Xiamen Municipal Science and Technology Program [3502Z20073015

Facile Deposition of Manganese Dioxide to Albumin-Bound Paclitaxel Nanoparticles for Modulation of Hypoxic Tumor Microenvironment To Improve Chemoradiation Therapy

Tumor microenvironment with hypoxia and excess hydrogen peroxide (H₂O₂) tremendously limits the effect of chemoradiation therapy of colorectal cancer. For the first time, we developed a facile method to deposit manganese dioxide (MnO₂) on the surface of albumin bound paclitaxel nanoparticles (ANPs-PTX) to obtain MnO₂-functioned ANPs-PTX (MANPs-PTX). In the tumor microenvironment, MANPs-PTX could consume excess hydrogen peroxide (H₂O₂) to produce abundant oxygen for tumor oxygenation and improve chemoradiation therapy. Meanwhile, the released Mn²⁺ from MANPs-PTX had excellent T₁ magnetic resonance imaging (MRI) performances for tumor detection. Notably, the obtained MANPs-PTX would be a promising theranostic agent and have potential clinical application prospects