ABSTRACT. Homocysteine (Hcy) is an independent risk factor of atherosclerosis through its involvement with the methionine cycle. In this study, we aimed to determine the blood vessel global methylation rate in Hcy-induced atherosclerosis in apolipoprotein-E-deficient (ApoE -/-) mice, and to explore the possible mechanism of this change in endothelial cells. ApoE -/-mice were divided into a hyperlipidemia (HLP) group, a hyperhomocysteinemia (HHcy) group, and an HHcy + folate + vitamin B12 (HHcy+FA+VB) group. Wild-type C57BL/6J mice were prepared as controls. Total Hcy, lipids, S-adenosylmethionine (SAM), and S-adenosylhomocysteine (SAH) contents in serum were measured with an automatic biochemistry analyzer and high-performance liquid chromatography. Methylation of B1 repetitive elements in blood vessels was tested using nested methylation-specific-polymerase chain reaction (nMS-PCR). Endothelial cells (ECs) were pretreated with Hcy or by adding FA and VB. Lectin-like oxidized LDL receptor-1 (LOX-1) expressions were determined by quantitative PCR, Western blot, and nMS-PCR. The HHcy group displayed severe HLP and HHcy. SAM and SAH contents were also elevated in the HHcy group compared with other groups. Methylation of B1 repetitive elements was significantly increased in the HHcy group (0.5050 ± 0.0182) compared to the HLP (0.5158 ± 0.0163) and control (0.5589 ± 0.0236) groups. mRNA and protein expressions of LOX-1 increased (0.2877 ± 0.0341, 0.6090 ± 0.0547), whereas methylation expression decreased (0.5527 ± 0.0148) after 100 µM Hcy stimulation in ECs. In conclusion, Hcy-induced atherosclerosis was closely associated with induced hypomethylation status in the blood vessel, and this process was partially mediated by LOX-1 DNA methylation.
Objective: A pivotal study was designed to investigate the effect of Hydroxy (HYC) Cu, Zn, and Mn on egg quality and laying performance of chickens under tropical conditions.Methods: A total of 1,260 Babcock White laying hens (20-wk-old) were randomly assigned to one of 4 treatments with 15 replicates of 21 hens each in a Randomized Complete Block Design. The birds were reared for 16 weeks and were fed the corn-soybean meal diets supplemented with one of the following mineral treatments: T1, inorganic (INO, 15 ppm CuSO4, 80ppm MnSO4 and 80 ppm ZnO); T2, Hydroxy-nutritional level (HYC-Nut, 15 ppm Cu, 80 ppm Mn, 80 ppm Zn from Hydroxy); T3, Hydroxy-Low (HYC-Low, 15 ppm Cu, 60 ppm Mn, 60 ppm Zn from Hydroxy); T4, Hydroxy plus inorganic (HYC+INO, 7.5 ppm HYC Cu+7.5 ppm CuSO4, 40 ppm HYC ZnO+40 ppm ZnSO4, 40 ppm HYC Mn+40 ppm MnSO4). The egg production was recorded daily, while the feed consumption, feed conversion ratio (FCR) and egg mass were determined at the end of each laying period. The egg quality parameters were assayed in eggs collected over 48 h in each laying period.Results: Overall, no significant effect of treatments was observed on percent egg production, egg weight and FCR (p>0.05). Feed intake was significantly lower in birds fed Hydroxy plus inorganic (p<0.05) diet. The supplementation of HYC-Low significantly increased the egg mass compared to the other treatments (p<0.05). HYC supplementation alone or in combination with INO elicited a positive effect on shell thickness, shell weight, shell weight per unit surface area, yolk colour, albumen and yolk index for a certain period (p<0.05), but not throughout the whole laying period.Conclusion: Dietary supplementation of HYC-Low (15-60-60 mg/kg) showed similar effects on production performance and egg quality characteristics in laying hens as compared to 15-80-80 mg/kg of Cu-Zn-Mn from inorganic sources. This indicates that sulphate based inorganic trace minerals can effectively be substituted by lower concentration of hydroxyl minerals.
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