Cloning, expression and regulation of Schizosaccharomyces pombe gene encoding thioltransferase.

The genomic DNA encoding thioltransferase was isolated from Schizosaccharomyces pombe using the polymerase chain reaction. The amplified DNA fragment was confirmed by Southern hybridization, completely digested with HindIII and BamHI, and then ligated into the yeast-Escherichia coli shuttle vector pRS316, which resulted in plasmid pEH1. The insert of plasmid pEH1 was transferred into the multi-copy vector YEp357 to generate plasmid pYEH1. The determined nucleotide sequence harbors an open reading frame consisting of four exons and three introns, which encodes a polypeptide of 101 amino acids with a molecular mass of 11261 Da. Thioltransferase activity was increased 1.6-fold in Saccharomyces cerevisiae containing plasmid pYEH1, and 1.8- and 2.7-fold in S. pombe containing plasmid pEH1 and pYEH1, respectively. The upstream sequence and the region encoding the N-terminal six amino acids were fused into promoterless beta-galactosidase gene of the shuttle vector YEp357R to generate the fusion plasmid pYEHR1. Synthesis of beta-galactosidase from the fusion plasmid was found to be enhanced by zinc and NO-generating S-nitroso-N-acetylpenicillamine.

Behavioral risk factors in an Amish community.

A representative sample of 400 Amish adults residing in Holmes County, Ohio, was interviewed about certain health risk characteristics and behaviors, using the Behavioral Risk Factor Survey (BRFS). For purposes of comparison, a representative sample of 773 non-Amish adults responded to the same survey by telephone interviews. In general, the Amish report lower rates of alcohol and tobacco consumption than their non-Amish counterparts. The Amish are less likely to salt their food and are more likely to take vitamin supplements, but do not differ from non-Amish in the consumption of "health foods." Amish men and women are less likely to be trying to lose weight than their non-Amish counterparts. Further, the Amish are less likely to engage in leisure-time physical activity or in exercise associated with attempts to lose weight or deal with hypertension. Amish women are less likely to use seat belts than non-Amish women, whereas men in both groups appear rather similar. Although some differences could be influenced by response biases, many are supported by less systematic observations of Old Order Amish societies. The patterns of health behavior reflect characteristics of Amish culture and may be responsible for certain favorable mortality rates among the Amish population.