preparation-pseudo-typed-h5-avian-influenza-viruses-with-calcium

Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity

Since 1996, A/goose/Guangdong/1/96-lineage highly pathogenic avian influenza (HPAI) H5 viruses have been causing flu outbreaks in poultry and wild birds. ...

Jilin Academy of Agricultural Sciences

In this study, we performed high throughput RNA sequencing at the primary bovine preadipocyte (Day-0), mid-differentiation (Day-4), and differentiated adipocyte (Day-9) stages in order to characterize the transcriptional events regulating differentiation and function. The preadipocytes were isolated from subcutaneous fetal bovine adipose tissues and were differentiated into mature adipocytes. The adipogenic characteristics of the adipocytes were detected during various stages of adipogenesis (Day-0, Day-4, and Day-9). We used RNA sequencing (RNA-seq) to investigate a comprehensive transcriptome information of adipocytic differentiation. Compared to the pre-differentiation stage (Day-0), 2510 genes were identified as differentially expressed genes (DEGs) at the mid-differentiation stage (Day-4). We found 2446 DEGs in the mature adipocytic stage relative to the mid-differentiation stage. Some adipogenesis-related transcription factors, CCAAT-enhancer-binding protein α (C/EBPα) and peroxisome proliferator-activated receptor γ (PPARγ) were differentially expressed at Day-0, Day-4, and Day-9. We further investigated the adipogenic function of 5-hydroxytryptamine receptor 2A (HTR2A) in adipogenesis. Overexpression of HTR2A stimulated the differentiation of preadipocytes, and knockdown of HTR2A had opposite effects. Furthermore, functional enrichment analysis of DEGs revealed that the PI3K-Akt signaling pathway was the significantly enriched pathway, and HTR2A regulated adipogenesis by activating or inhibiting phosphorylation of phospho-AKT (Ser473). In summary, the present study provides the first comparative transcription of various periods of adipocytes in cattle, which presents a solid foundation for further study into the molecular mechanism of fat deposition and the improvement of beef quality in cattle.

RNA-Seq Analysis Reveals a Positive Role of HTR2A in Adipogenesis in Yan Yellow Cattle.

The post-transcriptional events regulating expression of the adipogenesis associated Mth938 domain containing (AAMDC) protein are poorly understood. Here, we find that AAMDC expresses three isoforms due to alternative polyadenylation (APA) and alternative splicing (AS). Luciferase assay revealed that expression of the two isoforms with short and long 3'UTRs is differentially controlled. Further analyses showed that the short isoform displays higher translation efficiency and that miR-2428/664a reduces expression of the long isoform, indicating that APA-mediated shortening of the AAMDC 3'UTR renders the short isoform insusceptible to miRNA-mediated suppression due to loss of the binding sites for miR-2428/664a. Finally, we demonstrate that the short AAMDC isoform promotes bovine preadipocyte differentiation. Collectively, our findings indicate that AAMDC is post-transcriptionally regulated through APA and microRNAs.

Adipogenesis associated Mth938 domain containing (AAMDC) protein expression is regulated by alternative polyadenylation and microRNAs.

Pesticide chlorpyrifos (CPF) is a widespread environmental pollutant gaining attention as it is highly injurious to aquatic life. Although the toxicity of CPF is well characterized, but the mechanism of toxic response especially, the hepatotoxicity remained unclear. In this study, we performed integrated analysis, including micro-RNA (miRNA) and small RNA (sRNA) to analyze CPF exposure responding genes and enrichment pathways. A total of 23,742 expressed genes were detected and out of these expression levels of 1746 were changed significantly. Majority of them participated in protein biosynthesis, nucleotide binding, and oxidation-reduction activities. In extensive analysis of micro-RNA (miRNA) expression profiles by comparing CPF treated carp with control, we identified 214 novel miRNAs with CPM > 5 in at least one sample. The miRNAs have the same change in direction compared with overlapped mRNA pairs in upregulated genes, suggesting potential positive correlation. As a whole, we detected many differently expressed genes (DEGs) and miRNAs, which may be used as the biomarkers for the detection of CPF pollution in water and aquatic product safety. However, their functions are required to be deeply analyzed, especially more samples or time pointed data are needed to illustrate their concrete mechanism.

Translation, ribosome biogenesis, and oxidative damage caused by chlorpyrifos exposure to common carp (Cyprinus carpio L.) liver: application of combined RNA-seq with sRNA-seq in risk evaluation of environmental toxicant chlorpyrifos.

This work describes the characterization and genome annotation of the newly isolated lytic phage vB_SsoM_Z31 (referred to as Z31), isolated from wastewater samples collected in Dalian, China. Transmission electron microscopy revealed that phage Z31 belongs to the family Myoviridae, order Caudovirales. This phage specifically infects Shigella sonnei, Shigella dysenteriae, and Escherichia coli. The genome of the phage Z31 is an 89,355-bp-long dsDNA molecule with a G+C content of 38.87%. It was predicted to contain 133 ORFs and encode 24 tRNAs. No homologs of virulence factor genes or antimicrobial resistance genes were found in this phage. Based on the results of nucleotide sequence alignment and phylogenetic analysis, phage Z31 was assigned to the genus Felixounavirus, subfamily Ounavirinae.

Lichun Zhang

Institute of Animal Bio-technology

Jilin Academy of Agricultural Sciences

Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity