Novel SNP of the goat prolactin gene (PRL) associated with cashmere traits.

Concentrations of the single-chain polypeptide hormone prolactin (PRL) are associated with wool or cashmere traits, and its seasonal changes may determine patterns of enzymatic activity and may affect cashmere fibre growth and moult. So, the PRL gene is a potential candidate gene for cashmere traits in marker-assisted selection (MAS). In this paper, we report a novel missense single-nucleotide polymorphism (SNP) within the goat PRL gene in 1367 individuals by PCR-SSCP (polymerase chain reaction with single-strand conformation polymorphism) analysis and DNA sequencing. The novel X76049:g.576C>A mutation is confirmed by Eco24I PCR-RFLP (restriction fragment length polymorphism) analysis and causes a missense codon (Pro176Thr). The frequencies of allele C varied from 0.79 to 0.93 in 9 analysed goat populations. C allele was correlated with higher fibre length (P=0.014).

The crystal structure of ADP-L-glycero-D-mannoheptose 6-epimerase: catalysis with a twist.

BACKGROUND: ADP-L-glycero--mannoheptose 6-epimerase (AGME) is required for lipopolysaccharide (LPS) biosynthesis in most genera of pathogenic and non-pathogenic Gram-negative bacteria. It catalyzes the interconversion of ADP-D-glycero-D-mannoheptose and ADP-L-glycero-D-mannoheptose, a precursor of the seven-carbon sugar L-glycero-mannoheptose (heptose). Heptose is an obligatory component of the LPS core domain; its absence results in a truncated LPS structure resulting in susceptibility to hydrophobic antibiotics. Heptose is not found in mammalian cells, thus its biosynthetic pathway in bacteria presents a unique target for the design of novel antimicrobial agents. RESULTS: The structure of AGME, in complex with NADP and the catalytic inhibitor ADP-glucose, has been determined at 2.0 A resolution by multiwavelength anomalous diffraction (MAD) phasing methods. AGME is a homopentameric enzyme, which crystallizes with two pentamers in the asymmetric unit. The location of 70 crystallographically independent selenium sites was a key step in the structure determination process. Each monomer comprises two domains: a large N-terminal domain, consisting of a modified seven-stranded Rossmann fold that is associated with NADP binding; and a smaller alpha/beta C-terminal domain involved in substrate binding. CONCLUSIONS: The first structure of an LPS core biosynthetic enzyme leads to an understanding of the mechanism of the conversion between ADP-D-glycero--mannoheptose and ADP-L-glycero-D-mannoheptose. On the basis of its high structural similarity to UDP-galactose epimerase and the three-dimensional positions of the conserved residues Ser116, Tyr140 and Lys144, AGME was classified as a member of the short-chain dehydrogenase/reductase (SDR) superfamily. This study should prove useful in the design of mechanistic and structure-based inhibitors of the AGME catalyzed reaction.

Increasing binding of a transcription factor immediately downstream of the cap site of a cytomegalovirus gene represses expression.

A closely related family of ubiquitous DNA binding proteins, called MDBP, binds with high affinity to two 14 base pair (bp) sites within the human cytomegalovirus immediate early gene 1 (CMV IE1) enhancer and with low affinity to one site beginning 5 bp downstream of the CMV IE1 transcription start point (+5 site). Unlike several cap position downstream MDBP sites in mammalian genes, these MDBP sites do not require cytosine methylation for optimal binding. Mutation of one of the enhancer MDBP sites to prevent MDBP recognition modestly increased the function of a neighboring CREB binding site in a transient transfection assay in the context of one promoter construct. A much larger effect on reporter gene expression (a 10-fold reduction) was seen when the low affinity MDBP recognition sequence at position +5 was converted to a high affinity site in a plasmid containing the CMV IE1 promoter upstream of the reporter gene. Evidence that the increased binding of MDBP at the mutant site is largely responsible for the observed results was provided by transfection experiments with this high affinity MDBP +5 site re-mutated to a non-binding site and by in vitro transcription assay.

[503 cases of pallas pit viper bite with traditional Chinese medicine and Western medicine treatment].

In this paper 503 cases of pallas pit viper bite had been treated with the method of TCM-WM. Among the 503 patients 491 were cured (curative rate 97.61%), however, 12 patients were unfruitful (unfruitful rate 2.39%). The results showed that the poisoning degree of these cases had a positive correlation with patient's age, distance of snake teeth prints in wound, duration before admission, local and systemic clinical manifestation, degree of secondary infection and the number of the organ and system affected. In the treatment of the pallas pit viper bite, it was considered that should insisting the excluding poison and detoxifying methods, so as to cut off the absorption of the poison into the body and promote the discharge of the poison from the body, protect and improve the hepatic and renal functions, keep the balance of inter-circumstance. Attach importance to prevent and treat secondary infection, and raise the awareness to prevent and treat respiratory failure. All these mentioned above were considered the key points to increase the curative rate of the patients.

Active products of porcine trypsin after autolysis.

Porcine trypsin obtained from pancreas residues subsequent to insulin removal undergoes autolysis when subjected to chromatography and gives rise to new forms of autolyzed products with intra-chain split at bonds Lys145-Ala146 and Arg105-Val106. Incubation of 1% solutions of porcine trypsin either at pH 5.0 or at pH 9.1 induces autolysis to give active products involving one or two specific cleavages of bonds Lys145-Ala146 and Arg105-Val106 or Lys131-Ser132, as well as inactive degraded products. No evidence has been obtained that on autolysis of porcine trypsin, and active fragment with molecular weight lower than that of the parent molecule was identified. The active forms of autolyzed products of porcine trypsin have almost the same specific activity as the intact enzyme when assayed against BAEE. They are of the same molecular weight as the parent molecule. These findings indicate that that active forms of autolyzed products maintain the specific three-dimensional structure essential for the catalytic activity of the trypsin molecule.