MicrobPad MD: microbial pathogen diagnostic methods database.

Medical pathogens induce infections, illnesses and sometimes serious medical conditions in the infected hosts. Diagnosis of these pathogens is important for proper treatment and investigation of pathogenesis processes. Molecular techniques have been developed for facilitating accurate, sensitive and low-cost diagnosis of these pathogens. Based on these techniques, diagnostic devices have been developed for a number of pathogens. More devices are needed for comprehensive coverage of medical pathogens. To facilitate the development of these devices, a database with integrated information about diagnostic methods, targets, and primers/probes for the known bacterial, fungal and viral pathogens is needed. We developed the microbial pathogen diagnostic methods database MicrobPad MD (http://bidd.nus.edu.sg/group/MicrobPad/MicrobPad.asp or http://pha-bidd.nus.edu.sg/group/MicrobPad/MicrobPad.asp) to provide comprehensive information about the molecular diagnostic techniques, targets, primers/probes, detection procedures and conditions, and tested diagnostic accuracies and limit of diagnosis for 314 bacterial, fungal and viral species from 61 genera. While available, additional information such as pathogen strains and hosts, tissue distribution or habitats, cultivation methods, biochemical characteristics, virulence factors, morphology, diseases, symptoms, treatment and prevention methods are provided. Our Database covers 242 gene targets, 700 primers/probes, 340 virulence factors, and 261 diseases. Cross-links to the NCBI genome and SwissProt/UniProt databases are provided.

Assay, purification, and properties of bovine liver D-xylulokinase.

Convenient specific assay methods using commercially available materials were developed for the measurement of D-xylulokinase and D-ribulokinase activities. Using these assays, D-xylulokinase from bovine liver has been purified to homogeneity. Purification involved column chromatography on DEAE-cellulose, Sephadex G-100, Dyematrex Red A, and Superose 12. The enzyme preparation was obtained with final yields of 15-30% and had activity toward both D-xylulose and D-ribulose. The final specific activities ranged from 2.5 to 7.5 and 1.9 to 5.9 mol/min for the two substrates and Km values of 0.14 and 0.27 mM were obtained, respectively. For ATP, a Km value of 0.080 mM was obtained with either substrate. AMP, ADP, and cAMP inhibited competitively with respect to ATP; Ki values were 0.34, 0.35, and 1.0 mM, respectively. Xylulokinase thus prepared was a monomeric protein of 51,000 Da and had a pH optimum between 7.4 and 8.2. The kinetics of the enzyme do not support any significant regulation of flux through the enzyme by metabolite level changes.