One-step refolding and purification of disulfide-containing proteins with a C-terminal MESNA thioester

Background  

Expression systems based on self-cleavable intein domains allow the generation of recombinant proteins with a C-terminal thioester. This uniquely reactive C-terminus can be used in native chemical ligation reactions to introduce synthetic groups or to immobilize proteins on surfaces and nanoparticles. Unfortunately, common refolding procedures for recombinant proteins that contain disulfide bonds do not preserve the thioester functionality and therefore novel refolding procedures need to be developed.     

Factors associated with whole carcass condemnation rates in provincially-inspected abattoirs in Ontario 2001-2007: implications for food animal syndromic surveillance

Background  

Ontario provincial abattoirs have the potential to be important sources of syndromic surveillance data for emerging diseases of concern to animal health, public health and food safety. The objectives of this study were to: (1) describe provincially inspected abattoirs processing cattle in Ontario in terms of the number of abattoirs, the number of weeks abattoirs process cattle, geographical distribution, types of whole carcass condemnations reported, and the distance animals are shipped for slaughter; and (2) identify various seasonal, secular, disease and non-disease factors that might bias the results of quantitative methods, such as cluster detection methods, used for food animal syndromic surveillance.     

Cellulose-bound Peptide Arrays: Preparation and Applications

Spatial organization of metabolic enzymes may represent a general cellular mechanism to regulate metabolic flux. One recent example of this type of cellular phenomenon is the purinosome, a newly discovered multi-enzyme metabolic assembly that includes all of the enzymes within the de novo purine biosynthetic pathway. Our understanding of the components and regulation of purinosomes has significantly grown in recent years. This paper reviews the purine de novo biosynthesis pathway and its regulation, and presents the evidence supporting the purinosome assembly and disassembly processes under the control of G-protein-coupled receptor (GPCR) signaling. This paper also discusses the implications of purinosome and GPCR regulation in drug discovery.     

Adsorption and Degradation of Metolachlor and Metribuzin in a No-Till System Under Three Winter Crop Covers

The objective of this study was to quantify adsorption and degradation of metolachlor [2-chloro-N-(2-ethyl-6-methylphenyl)-N-(2-methoxy-1 -methylethyl) acetamide] and metribizun [4-amino-6-(1,1 -dimethylethyl)-3-(methylthio)-1,2,4-trazine-5(4H)-one] in a soil planted to winter covers clover (Trifolium sp.), vetch (Vicia villosa), and wheat (Triticum aestivum). Surface soil samples (0 to 5?cm) from Memphis silt loam (fine-silty, mixed, thermic Typic Hapludalf) were collected and equilibrated with herbicide at initial concentrations ranging from 0 to 20?mg L^?1 that were then applied for a batch experiment. Soils were treated either with a single herbicide or a mixture of the two herbicides. For the degradation experiments, herbicides at a concentration of 10?mg kg^?1 soil were applied and incubated for 21?d at ～23°C. Metolachlor and metribuzin adsorptions were described by the Freundlich isotherm. Average Freundlich distribution coefficient (K_f) for metolachlor was significantly higher (p≥0.05) than that of metribuzin in soils under the three crop covers irrespective of method of application. The K_f for metolachlor ranged from 18.38 to 11.18?L kg^?1, and K_f for metribuzin ranged from 1.80 to 0.93?L kg^?1. Average normalized distribution coefficient (K_oc) for metolachlor was significantly higher (p≥0.05) than average K_oc for metribuzin irrespective of crop cover. After 21 days of incubation, average half-life of metolachlor across soil under the three crop covers was significantly higher than the average half-life of metribuzin (p≥0.05). Half-life values ranged from 20.6 to 24.9 days for metolachlor, and 4.4 to 12.4 days for metribuzin. In soils treated with metribuzin, the half-life was highest for soil under wheat and lowest for soil under clover (p≥0.05).     

Incidence of Infectious Drug Resistance Among Lactose-Fermenting Bacteria Isolated from Raw and Treated Sewage

Raw and treated sewage samples were examined for antibiotic-resistant, lactose-fermenting bacteria. Approximately 1% of the total lactose-fermenting bacteria were multiply resistant. Of these organisms, 50% were capable of transferring all or part of their resistance to a drug-sensitive recipient. Only 43% of those isolated on media containing a single antibiotic were capable of resistance transfer, whereas 57% of those recovered on multiple antibiotic plates transferred resistance. R factors conferring resistance to chloramphenicol, streptomycin, and tetracycline; streptomycin and tetracycline; and ampicillin, streptomycin, and tetracycline accounted for 22, 19, and 15%, respectively, of those identified. The data indicate a significant level of infectious drug resistance among the intestinal bacteria of the urban population.