Effects of dietary supplementation of vitamins D(3) and E on quality characteristics of pigs and longissimus muscle antioxidative capacity.

The effects of addition of vitamin D(3) and vitamin E to pig diets on blood plasma calcium concentration, meat quality (longissimus muscle) and antioxidative capacity were investigated. Two treatments consisted of supplementation with vitamin D(3) (500,000IU/d) for 5 days separately (group D) and a combination of vitamin E (500mg α-tocopheryl acetate/kg diet) for 30 days and vitamin D(3) (500,000IU/d) for 5 days (group D+E) to growing-finishing pigs before slaughter. Pigs fed with vitamin D(3) had higher (P<0.01) plasma calcium concentration compared with control pigs. Dietary supplementation of vitamin E significantly (P<0.05) increased the concentration of α-tocopherol in meat (longissimus muscle). Vitamin D(3) supplementation resulted in higher (P=0.07) a(∗) values of loin chops at 5 days of storage. Vitamin D(3) and vitamin E supplementation did not affect other meat quality characteristics or tenderness (quantified by Warner-Bratzler shear force). Antioxidative capacity (measured as MDA production after incubation of longissimus muscle homogenates with Fe(2+)/ascorbate) was improved by vitamin E and partly by vitamin D(3) supplementation.

Rapid and reliable genotyping for the Toll-like receptor 4 A896G polymorphism using fluorescence-labeled hybridization probes in a real-time polymerase chain reaction assay.

BACKGROUND: The Toll-like receptor 4 (TLR4) is involved in immune response to endotoxin as well as the pathogenesis of atherosclerosis. The TLR4 gene was shown to carry a single-nucleotide polymorphism (A896G). We developed a rapid-cycle polymerase chain reaction (PCR) which allows for rapid genotyping and, therefore, may be useful in clinical risk assessment. METHODS: Fluorescently labeled oligonucleotide hybridization probes were designed for the LightCycler instrument. A PCR product was generated in a single reaction followed by melting point analysis. Ninety-three German Caucasians were genotyped. The interleukin-1beta (IL-1beta) response to endotoxin was assessed after whole blood stimulation with endotoxin according to TLR4 genotypes. RESULTS: The method suggested by us is a time-saving technique requiring no additional manual steps. Frequencies of the A and G allele were 0.95 and 0.05, respectively. The study population was in Hardy-Weinberg equilibrium. The specificity of the method was confirmed by sequencing. The IL-1beta levels were lower in carriers of the G allele. CONCLUSIONS: This PCR assay is a rapid and reliable technique for TLR4 genotyping.