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Piglet diarrhea is a swine disease responsible for serious economic impacts in the pig industry. Clostridium perfringens beta2 toxin (CPB2), which is a major toxin of C. perfringens type C, may cause intestinal diseases in many domestic animals. N6-methyladenosine (m6A) RNA methylation plays critical roles in many immune and inflammatory diseases in livestock and other animals. However, the role of m6A methylation in porcine intestinal epithelial (IPEC-J2) cells exposed to CPB2 has not been studied. To address this issue, we treated IPEC-J2 cells with CPB2 toxin and then quantified methylation-related enzyme expression by RT-qPCR and assessed the m6A methylation status of the samples by colorimetric N6-methyladenosine quantification. The results showed that the methylation enzymes changed to varying degrees while the m6A methylation level increased (p < 0.01). On this basis, we performed N6-methyladenosine sequencing (m6A-seq) and RNA sequencing (RNA-seq) to examine the detailed m6A modifications and gene expression of the IPEC-J2 cells following CPB2 toxin exposure. Our results indicated that 1,448 m6A modification sites, including 437 up-regulated and 1,011 down-regulated, differed significantly between CPB2 toxin exposed cells and non-exposed cells (p < 0.05). KEGG pathway analysis results showed that m6A peaks up-regulated genes (n = 394) were mainly enriched in cancer, Cushing syndrome and Wnt signaling pathways, while m6A peaks down-regulated genes (n = 920) were mainly associated with apoptosis, small cell lung cancer, and the herpes simplex virus 1 infection signaling pathway. Furthermore, gene expression (RNA-seq data) analysis identified 1,636 differentially expressed genes (DEGs), of which 1,094 were up-regulated and 542 were down-regulated in the toxin exposed group compared with the control group. In addition, the down-regulated genes were involved in the Hippo and Wnt signaling pathways. Interestingly, the combined results of m6A-seq and RNA-seq identified genes with up-regulated m6A peaks but with down-regulated expression, here referred to as "hyper-down" genes (n = 18), which were mainly enriched in the Wnt signaling pathway. Therefore, we speculate that the genes in the Wnt signaling pathway may be modified by m6A methylation in CPB2-induced IPEC-J2 cells. These findings provide new insights enabling further exploration of the mechanisms underlying piglet diarrhea caused by CPB2 toxin. [ABSTRACT FROM AUTHOR]

The effects of rice bran and corn on ensiling characteristics of potato vines were investigated. Ruminal fermentation of potato vine silage was measured using a rumen simulation technique (Rusitec). Potato vines were treated (fresh weight basis) for ensiling without additives (control), with 15% rice bran and with 15% rice bran plus 3% corn, 5% corn, or 7% corn, respectively. Silage with 5% corn was chosen for a ruminal fermentation trial, based on the results of ensiling quality. Comparison of rumen fermentation between two silage (silage with 5% corn and whole corn silage) diets was conducted using a Rusitec apparatus. Adding rice bran and corn improved the potato vine silage quality (P <.05). Compared with whole corn silage, potato vine silage diet had lower (P <.05) methane production, ruminal pH, acetate, and acetate to propionate ratio and higher (P <.05) total volatile fatty acids, total nitrogen (N), ammonia-N, and microbial-N. These findings indicated that rice bran and corn improved ensiling fermentation characteristics of potato vines, and that additive-treated potato vine silage had no adverse effects on rumen fermentation and could improve the energy and N utilisation and alleviate the methane pollution of the rumen compared with whole corn silage. [ABSTRACT FROM AUTHOR]

Introduction: The aim of this present work was to investigate the mechanism of the microRNA (miR)-216a-5p/FASL axis in mice with acute kidney injury (AKI)., Methods: Mice kidney ischemia/reperfusion (I/R) injury was used as AKI models in this study. I/R mice were injected with miR-216a-5p- and FASL-related constructs to investigate potential mechanisms of kidney protection. Kidney function, inflammation, oxidative stress, and kidney cell apoptosis were assessed after 24 h of reperfusion. In vitro, the hypoxia-reoxygenation (H/R) model was used with kidney tubular epithelial cells (TECs) to mimic kidney I/R injury. H/R-treated TECs were transfected with miR-216a-5p- and FASL-related constructs to detect cell viability, inflammation, and oxidative stress. MiR-216a-5p and FASL expression levels in mouse kidney tissues and in H/R-treated TECs were detected., Results: MiR-216a-5p was downregulated and FASL was upregulated in kidney tissues of I/R mice and H/R-treated TECs. Upregulating miR-216a-5p attenuated kidney cell apoptosis and the damage of kidney function, and reduced inflammatory factor levels and oxidative stress response in kidney tissues of I/R mice. Upregulating miR-216a-5p advanced cell viability and reduced inflammatory factor levels and oxidative stress response in H/R-treated TECs. Downregulation of FASL effectively reversed the influences of downregulation of miR-216a-5p on kidney injury in mice and kidney TEC survival., Conclusion: Our study reveals that miR-216a-5p reduces I/R-induced pathological kidney damage in AKI via suppressing FASL., (© 2024 The Author(s). Published by S. Karger AG, Basel.)

Piglet diarrhea is a swine disease responsible for serious economic impacts in the pig industry. Clostridium perfringens beta2 toxin (CPB2), which is a major toxin of C. perfringens type C, may cause intestinal diseases in many domestic animals. N 6 -methyladenosine (m 6 A) RNA methylation plays critical roles in many immune and inflammatory diseases in livestock and other animals. However, the role of m 6 A methylation in porcine intestinal epithelial (IPEC-J2) cells exposed to CPB2 has not been studied. To address this issue, we treated IPEC-J2 cells with CPB2 toxin and then quantified methylation-related enzyme expression by RT-qPCR and assessed the m 6 A methylation status of the samples by colorimetric N 6 -methyladenosine quantification. The results showed that the methylation enzymes changed to varying degrees while the m 6 A methylation level increased ( p < 0.01). On this basis, we performed N 6 -methyladenosine sequencing (m 6 A-seq) and RNA sequencing (RNA-seq) to examine the detailed m 6 A modifications and gene expression of the IPEC-J2 cells following CPB2 toxin exposure. Our results indicated that 1,448 m 6 A modification sites, including 437 up-regulated and 1,011 down-regulated, differed significantly between CPB2 toxin exposed cells and non-exposed cells ( p < 0.05). KEGG pathway analysis results showed that m 6 A peaks up-regulated genes ( n = 394) were mainly enriched in cancer, Cushing syndrome and Wnt signaling pathways, while m 6 A peaks down-regulated genes ( n = 920) were mainly associated with apoptosis, small cell lung cancer, and the herpes simplex virus 1 infection signaling pathway. Furthermore, gene expression (RNA-seq data) analysis identified 1,636 differentially expressed genes (DEGs), of which 1,094 were up-regulated and 542 were down-regulated in the toxin exposed group compared with the control group. In addition, the down-regulated genes were involved in the Hippo and Wnt signaling pathways. Interestingly, the combined results of m 6 A-seq and RNA-seq identified genes with up-regulated m 6 A peaks but with down-regulated expression, here referred to as "hyper-down" genes ( n = 18), which were mainly enriched in the Wnt signaling pathway. Therefore, we speculate that the genes in the Wnt signaling pathway may be modified by m 6 A methylation in CPB2-induced IPEC-J2 cells. These findings provide new insights enabling further exploration of the mechanisms underlying piglet diarrhea caused by CPB2 toxin., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Zhang, Yang, Yang, Gao, Luo, Huang, Yan, Wang, Wang, Xie, Zhang and Gun.)