There was no difference between towns in plasma folate or B12 levels but the conventional inverse relationship with plasma homocysteine was seen. Women in both towns had significantly lower plasma homocysteine than men. Plasma homocysteine levels were significantly higher in men from the coronary artery disease high risk town of Dewsbury than in the low coronary artery disease risk town of Maidstone (p<0.001), but not in women. ![]() Elderly men and women from two towns, of contrasting risk of developing of coronary artery disease, Dewsbury and Maidstone, were examined for plasma homocysteine levels and genotyped for the C677T and the A1298C polymorphisms in the MTHFR gene. This interaction between CBS genotype and MTHFR and MS genotypes points to a key role of the CBS transsulphuration pathway in the metabolism of homocysteine that may be particularly important in subjects with low dietary folate. Age, folate, B12 and smoking explained 24.2% of the variance while the three genotypes combined and with interaction terms explained an additional 3.8%. Furthermore there was an interaction between MTHFR and CBS genotypes and between MS and CBS genotypes. The raising effect associated with homozygosity for the 677T allele was greater in men in the lowest quartile of folate (interaction p=0.02). Individuals homozygous for the MTHFR 677T allele had highest median homocysteine levels when compared to the other two genotypes (p<0.001). ![]() Genotype at all three loci were associated with differences in plasma homocysteine level. Healthy middle aged men, from eight general practices across Britain (Northwick Park Heart Study II) were examined for plasma homocysteine levels and genotyped for the C677T polymorphism in the MTHFR gene, the 68 bp insertion polymorphism in exon 8 of the CBS gene and the A2756G polymorphism in the MS gene. There are three main enzymes involved in homocysteine metabolism Methylenetetrahydrofolate reductase (MTHFR), that is involved in the remethylation pathway, Methionine synthase (MS) that is also involved in the remethylation pathway and Cystathionine beta synthase (CBS) that is involved in the transsulphuration pathway. Mutations in the enzymes regulating homocysteine metabolism have been associated with elevated plasma homocysteine levels. Mild hyperhomocysteinaemia occurs commonly in individuals in the general population and has been associated with increased risk of developing atherosclerosis and coronary artery disease. This method is rapid, cost-effective, and suitable for large-scale sample screening as compared with the phylogenetic analysis of the SSU rDNA gene sequences. The SSCP fingerprint patterns were related to the genus-level taxonomy of dictyostelids, but the fingerprint dendrogram did not reflect the deep phylogeny. We also tested the optimized SSCP procedure with 73 Thai isolates of dictyostelid that had the SSU rDNA gene sequences published. The optimized SSCP condition required gel purification of the SSCP amplicons followed by electrophoresis using a 9% polyacrylamide gel under 4 C. Here, we designed a set of PCR-single-strand conformation polymorphism (SSCP) primers and optimized the SSCP fingerprint method for the screening of dictyostelids. Genetic diversity study of these organisms solely relies on molecular phylogenetics of the SSU rDNA gene, which is not ideal for large-scale genetic diversity study. Dictyostelid social amoebae are a highly diverse group of eukaryotic soil microbes that are valuable resources for biological research.
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