Comprehensive evaluation of treating drinking water for laying hens using slightly acidic electrolyzed water.

Slightly acidic electrolyzed water (SAEW) is well-known for its highly potent antibacterial properties and safe residue-free nature. In this study, a comprehensive evaluation was conducted on 2 disinfection methods for waterline cleaning in poultry houses: (1) continuously add SAEW into the waterline and (2) the conventional waterline disinfection method, which includes regular use of high-concentration chemical disinfectant for soaking the waterline and flushing with water. The evaluation focused on the effects of these methods on bacteria levels in laying hens' drinking water, the fecal normal rate of laying hens, egg quality, as well as the economic costs and water footprint associated with each method. The results show that the inhibition rate of the control group was 52.45% to 80.36%, which used 1500 mg/L sodium dichloroisocyanurate (DCCNa) for soaking and then flushing with water. The bacterial levels in the waterline returned to pre-treatment levels 26 h after cleaning. However, the experimental group with an available chlorine concentration (ACC) of 0.3 mg/L SAEW showed a higher inhibition rate (99.90%) than the control group (P < 0.05) and exhibited a sustained antimicrobial effect. Regarding eggshell thickness, eggshell strength, and Haugh units of the egg, there were no significant differences between the experimental and control groups. However, the experimental group had higher egg weight and darker yolk color (P < 0.05) than those of the control group. Besides, the experimental group exhibited a higher fecal normal rate and a lower water footprint than those of the control group. Hence, SAEW represents a favorable choice for disinfecting drinking water in poultry houses due to its ease of preparation, lack of residue, energy efficiency, and efficient antibacterial properties. To ensure adequate sanitation, it is recommended to incorporate SAEW with an ACC of 0.3 mg/L into the daily management of the drinking water system for laying hens.

LSD1 knockdown confers protection against osteoclast formation by reducing histone 3 lysine 9 monomethylation and dimethylation in ITGB3 promoter.

ITGB3, an osteoclast marker, is involved in osteoclast formation. Nevertheless, its related mechanism remains poorly characterized. Herein, this study examines the mechanisms affecting osteoclast formation with the involvement of ITGB3. Osteoclast formation was induced with macrophage colony-stimulating factor (M-CSF) and receptor activator of nuclear factor-kappa B ligand (RANKL), followed by measurement of the mRNA and protein expression of ITGB3 and LSD1. After gain- and loss-of-function assays, cell viability and the expression of osteoclast marker genes (NFATc1, ACP5, and CTSK) were assessed, and osteoclast formation was evaluated with TRAP staining. ChIP assays were used to examine histone 3 lysine 9 (H3K9) monomethylation (H3K9me1) and H3K9 dimethylation (H3K9me2) modifications and LSD1 protein enrichment in the ITGB3 promoter. During osteoclast formation, ITGB3 and LSD1 were gradually augmented. Knockdown of LSD1 or ITGB3 curbed cell viability, the expression of osteoclast marker genes, and osteoclast formation. Moreover, overexpression of ITGB3 nullified the suppressive impact of LSD1 knockdown on osteoclast formation. Mechanistically, LSD1 promoted ITGB3 expression by reducing H3K9 levels in the ITGB3 promoter. LSD1 enhanced ITGB3 expression by decreasing H3K9me1 and H3K9me2 levels in ITGB3 promoter to boost osteoclast formation.

microRNA-25-3p suppresses osteogenic differentiation of BMSCs in patients with osteoporosis by targeting ITGB3.

This study was conducted to investigate the impact of the microRNA (miR)-25-3p/ITGB3 axis on the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) from patients with osteoporosis (OP). BMSCs isolated from the bone marrow of healthy controls and OP patients were identified by flow cytometry, in which ITGB3 mRNA and miR-25-3p expression was detected by RT-qPCR and ITGB3, Runx2, OPN, ALP, and OSX protein expression by western blot. The binding between ITGB3 and miR-25-3p was assessed by dual-luciferase reporter gene and Ago2-RIP assays. BMSC osteogenic differentiation was observed by alizarin red staining and ALP activity. The differentiation of BMSCs to adipocytes and chondrocytes was measured by oil red O staining and alcian blue staining, respectively. BMSCs were successfully isolated from the bone marrow of healthy controls (normal-BMSCs) and OP patients (OP-BMSCs). ITGB3, Runx2, OPN, ALP, and OSX expression was poorer and miR-25-3p expression was higher in OP-BMSCs than in normal-BMSCs. Mechanistically, ITGB3 was negatively targeted by miR-25-3p. After osteogenic, adipogenic, and chondrogenic differentiation of BMSCs were successfully induced, adipogenic differentiation increased and osteogenic and chondrogenic differentiation decreased in OP-BMSCs compared with normal-BMSCs. Overexpression of ITGB3 facilitated mineralized nodule formation and elevated ALP activity and Runx2, OPN, and ALP expression in OP-BMSCs. miR-25-3p upregulation diminished mineralized nodule formation, ALP activity, and Runx2, OPN, and ALP expression in OP-BMSCs and normal-BMSCs, which was annulled by additional ITGB3 overexpression. miR-25-3p targets ITGB3, thereby suppressing osteogenic differentiation of BMSCs from OP patients.