Molecular analysis of the distribution and phylogeny of the soxB gene among sulfur-oxidizing bacteria - evolution of the Sox sulfur-oxidizing enzyme system

The soxB gene encodes the SoxB component of the periplasmic thiosulfate-oxidizing Sox enzyme complex, which has been proposed to be widespread among the various phylogenetic groups of sulfur-oxidizing bacteria (SOB) that convert thiosulfate to sulfate with and without the formation of sulfur globules as intermediate. Indeed, the comprehensive genetic and genomic analyses presented in the present study identified the soxB gene in 121 phylogenetically and physiologically divergent SOB, including several species for which thiosulfate utilization has not been reported yet. In first support of the previously postulated general involvement of components of the Sox enzyme complex in the thiosulfate oxidation process of sulfur-storing SOB, the soxB gene was detected in all investigated photo- and chemotrophic species that form sulfur globules during thiosulfate oxidation (Chromatiaceae, Chlorobiaceae, Ectothiorhodospiraceae, Thiothrix, Beggiatoa, Thiobacillus, invertebrate symbionts and free-living relatives). The SoxB phylogeny reflected the major 16S rRNA gene-based phylogenetic lineages of the investigated SOB, although topological discrepancies indicated several events of lateral soxB gene transfer among the SOB, e.g. its independent acquisition by the anaerobic anoxygenic phototrophic lineages from different chemotrophic donor lineages. A putative scenario for the proteobacterial origin and evolution of the Sox enzyme system in SOB is presented considering the phylogenetic, genomic (sox gene cluster composition) and geochemical data

Export of functional Streptomyces coelicolor alditol oxidase to the periplasm or cell surface of Escherichia coli and its application in whole-cell biocatalysis

Streptomyces coelicolor A3(2) alditol oxidase (AldO) is a soluble monomeric flavoprotein in which the flavin cofactor is covalently linked to the polypeptide chain. AldO displays high reactivity towards different polyols such as xylitol and sorbitol. These characteristics make AldO industrially relevant, but full biotechnological exploitation of this enzyme is at present restricted by laborious and costly purification steps. To eliminate the need for enzyme purification, this study describes a whole-cell AldO biocatalyst system. To this end, we have directed AldO to the periplasm or cell surface of Escherichia coli. For periplasmic export, AldO was fused to endogenous E. coli signal sequences known to direct their passenger proteins into the SecB, signal recognition particle (SRP), or Twin-arginine translocation (Tat) pathway. In addition, AldO was fused to an ice nucleation protein (INP)-based anchoring motif for surface display. The results show that Tat-exported AldO and INP-surface-displayed AldO are active. The Tat-based system was successfully employed in converting xylitol by whole cells, whereas the use of the INP-based system was most likely restricted by lipopolysaccharide LPS in wild-type cells. It is anticipated that these whole-cell systems will be a valuable tool for further biological and industrial exploitation of AldO and other cofactor-containing enzymes.

Wellness in the Face of Frailty Among Older Adults in First Nations Communities

OBJECTIVES: First Nations people report high levels of wellness despite high rates of chronic illness. Our goal was to understand the factors associated with wellness among First Nations adults in Ontario who were considered frail. METHODS: Using the First Nations Regional Health Survey, we created a profile of First Nations adults (aged 45+) who were categorized as frail (weighted sample size = 8121). We used multivariate logistic regression to determine associations between wellness (as measured by self-reported physical, emotional, mental, and spiritual balance) and determinants of health. RESULTS: Rates of reported wellness were high among those who were frail, ranging from 56.7% reporting physical balance to 71.6% reporting mental balance. Three key elements were associated with wellness: the availability of resources, individual lifestyle factors, and cultural connection and identity. DISCUSSION: Our findings provide a profile of strength and wellness among older First Nations adults living with frailty

The Aquatic Toxicity of Organic Compounds to Embryo-Larval Stages of Fish and Amphibians

Aquatic toxicity tests were conducted on 11 organic compounds considered hazardous to water resources. The toxicity of each compound was evaluated using embryo-larval stages of two to eight fish and amphibian species. Exposure was initiated at fertilization and maintained through 4 days posthatching. The animal test species exhibited varying degrees of sensitivity to the selected toxicants. Combined frequencies for mortality and teratogenesis at 4 days posthatching gave LC50 ranges of 3.66 to 8.25 mg/L for benzene, 1.16 to 22.42 mg/L for carbon tetrachloride, 0.11 to 1.20 mg/L for chlorobenzene, 2.03 to \u3e 68 mg/L for chloroform, 3.01 to 5.56 mg/L for 1,2-dichlorobenzene, 2.54 to .34 mg/L for 1,2-dichloroethane, 13.16 to \u3e 48 mg/L for methylene chloride, 0.002 to 0.64 mg/L for nitrobenzene, 0.04 to .32 mg/L for phenol, 0.02 to 0.85 mg/L for toluene, and 3.53 to 3.77 mg/L for m-xylene. The species which exhibited the greatest susceptibility to organic compounds were the rainbow trout, Rana pipiens, and Rana temporaria. The more sensitive amphibian species generally were those which normaly are restricted to aquatic or moist terrestrial habitats, whereas the more tolerant amphibians included those semi-aquatic and terrestrial species which appear to be more broadly adapted ecologically. Of the 11 test compounds, nitrobenzene, toluene, chlorobenzene, and phenol were the most toxic. The least toxic organics included dichloroethane and methylene chloride. For three chlorinated alkanes, including methylene chloride (CH2Cl2), chloroform (CHCl3), and carbon tetrachloride (CCl4), toxicity was found to 1ncrease with the degree of chlorination. Concerning several aromatic hydrocarbons, benzene always was found to be less toxic than its monosubstituted analogs. Toxicity of the 11 compounds was further evaluated by calculating toxicant concentrations which produced embryo-larval mortality and/or teratogenesis at frequencies of 10% (LC10) and 1% (LC1). The LC values, used to estimate toxicity thresholds, ranged from \u3c 0.l for nitrobenzene to 69.9 μg/L for methylene chloride. A limited number of toxicity tests were performed to determine whether embryo-larval bioassays are suitable to assess effects of transitory chemical exposures, such as those resulting from intermittent discharges or accidental spills of chemicals into water resources. Results indicated that Rana pipiens embryos were sufficiently sensitive to quantify effects produced by short-term exposures to chloroform. Animals tested during the earliest embryonic stage appeared to be less tolerant than organisms exposed later in development

A DIRECT LATERAL APPROACH TO THE LATERAL COMPARTMENT OF THE KNEE JOINT

A description is given of a direct approach to the lateral compartment of the knee with the joint fully flexed. This approach has been found useful for excision of cysts of the lateral cartilage, but is also applicable to excision of the lateral cartilage for tear. </jats:p