Characterization of Molybdenum-Free Nitrate Reductase from Haloalkalophilic Bacterium Halomonas sp. Strain AGJ 1-3

Nitrate reductase from the haloalkalophilic denitrifying bacterium Halomonas sp. strain AGJ 1-3 was isolated and purified to homogeneity. The isolated enzyme belongs to a novel family of molybdenum-free nitrate reductases. It presents as a 130-140 kD monomeric protein with specific activity of 250 micromol/min per mg protein. The enzyme reduces not only nitrate, but also other anions, thus showing polyoxoanion reductase activity. Enzyme activity was maximal at pH 7.0 and 70-80 degrees C.     

The enlarged population of marginal zone/CD1d(high) B lymphocytes in nonobese diabetic mice maps to diabetes susceptibility region Idd11

The NOD mouse is an important experimental model for human type 1 diabetes. T cells are central to NOD pathogenesis, and their function in the autoimmune process of diabetes has been well studied. In contrast, although recognized as important players in disease induction, the role of B cells is not clearly understood. In this study we characterize different subpopulations of B cells and demonstrate that marginal zone (MZ) B cells are expanded 2- to 3-fold in NOD mice compared with nondiabetic C57BL/6 (B6) mice. The NOD MZ B cells displayed a normal surface marker profile and localized to the MZ region in the NOD spleen. Moreover, the MZ B cell population developed early during the ontogeny of NOD mice. By 3 wk of age, around the time when autoreactive T cells are first activated, a significant MZ B cell population of adult phenotype was found in NOD, but not B6, mice. Using an F2(B6 x NOD) cross in a genome-wide scan, we map the control of this trait to a region on chromosome 4 (logarithm of odds score, 4.4) which includes the Idd11 and Idd9 diabetes susceptibility loci, supporting the hypothesis that this B cell trait is related to the development of diabetes in the NOD mouse.     

Genealogic analysis of the resistance of winter wheat to common bunt

Comparative genealogical analysis of North American (the United States and Canada) and Eastern European (Russia and Ukraine) winter wheat cultivars resistant and susceptible to common bunt has been performed. Analysis of variance applied to North American wheats has demonstrated that resistant and susceptible cultivars significantly differ from each other with respect to the contributions of common ancestors. The contributions of Oro (Bt4 and Bt7), Rio (Bt6), White Odessa (Bt1), and Florence (Bt3) to the resistant cultivars are significantly higher than their contributions to the susceptible ones. This demonstrates that the use of these resistance donors in wheat breeding for several decades has been effective. The contribution of PI-178383 (Bt8, Bt9, and Bt10) is considerably higher in the group of resistant cultivars bred after 1965. The mean contributions of Federation (Bt7) and Nebred (Bt4) are significantly higher in the group of resistant cultivars obtained before 1965; however, the differences in the contributions of these donors between new resistant and susceptible cultivars became nonsignificant. Among the Russian and Ukrainian cultivars, there are differences between groups of resistant and susceptible cultivars from different regions determined by the differences between the regional populations of the pathogen in racial composition. In the northern region, the contributions of the wheat grass (Agropyron glaucum) and the rye cultivar Eliseevskaya are significantly higher in the resistant cultivars; in the southern region, a local cultivar of the Odessa oblast is the prevalent resistant cultivar. In addition, cultivar Yaroslav Emmer is likely to be effective in the northern region; and foreign sources (Oro, Florence, Federation, and Triticum timopheevii), in the southern region. Very few sources of vertical resistance to common bunt are used for winter wheat breeding in Russia and Ukraine. The decrease in genetic diversity in favor of a few identical genes may cause adequate changes in the pathogen population and subsequent proliferation of the pathogen on the genetically identical substrate. A new interpretation of the resistance of line Lutescens 6028 as a source of new genes, Bt12 and Bt13, is suggested. Both genealogical and segregation analyses have shown that the genes determining the resistance of this line may be identical to those described earlier (Bt1, Bt3, Bt4, Bt6, and Bt7); and the high resistance of this line is determined by a combination of these genes.     

Functional and Morphological Adaptation to Peptidoglycan Precursor Alteration in Lactococcus lactis

Cell wall peptidoglycan assembly is a tightly regulated process requiring the combined action of multienzyme complexes. In this study we provide direct evidence showing that substrate transformations occurring at the different stages of this process play a crucial role in the spatial and temporal coordination of the cell wall synthesis machinery. Peptidoglycan substrate alteration was investigated in the Gram-positive bacterium Lactococcus lactis by substituting the peptidoglycan precursor biosynthesis genes of this bacterium for those of the vancomycin-resistant bacterium Lactobacillus plantarum. A set of L. lactis mutant strains in which the normal d-Ala-ended precursors were partially or totally replaced by d-Lac-ended precursors was generated. Incorporation of the altered precursor into the cell wall induced morphological changes arising from a defect in cell elongation and cell separation. Structural analysis of the muropeptides confirmed that the activity of multiple enzymes involved in peptidoglycan synthesis was altered. Optimization of this altered pathway was necessary to increase the level of vancomycin resistance conferred by the utilization of d-Lac-ended peptidoglycan precursors in the mutant strains. The implications of these findings on the control of bacterial cell morphogenesis and the mechanisms of vancomycin resistance are discussed.     

Electrostatic properties of promoter recognized by E. coli RNA polymerase Esigma70

A comparative analysis of electrostatic patterns for 359 sigma70-specific promoters and 359 nonpromoter regions on electrostatic map of Escherichia coli genome was carried out. It was found that DNA is not a uniformly charged molecule. There are some local inhomogeneities in its electrostatic profile which correlate with promoter sequences. Electrostatic patterns of promoter DNAs can be specified due to the presence of some distinctive motifs which differ for different promoter groups and may be involved as signal elements in differential recognition of various promoters by the enzyme. Some specific electrostatic elements which are responsible for modulating promoter activities due to ADP-ribosylation of RNA polymerase alpha-subunit were found in far upstream regions of T4 phage early promoters and E. coli ribosomal promoters.     

Multiple-Locus Sequence Typing Analysis of Bacillus cereus and Bacillus thuringiensis Reveals Separate Clustering and a Distinct Population Structure of Psychrotrophic Strains

We used multilocus sequence typing (MLST) to characterize phylogenetic relationships for a collection of Bacillus cereus group strains isolated from forest soil in the Paris area during a mild winter. This collection contains multiple strains isolated from the same soil sample and strains isolated from samples from different sites. We characterized 115 strains of this collection and 19 other strains based on the sequences of the clpC, dinB, gdpD, panC, purF, and yhfL loci. The number of alleles ranged from 36 to 53, and a total of 93 allelic profiles or sequence types were distinguished. We identified three major strain clusters—C, T, and W—based on the comparison of individual gene sequences or concatenated sequences. Some less representative clusters and subclusters were also distinguished. Analysis of the MLST data using the concept of clonal complexes led to the identification of two, five, and three such groups in clusters C, T, and W, respectively. Some of the forest isolates were closely related to independently isolated psychrotrophic strains. Systematic testing of the strains of this collection showed that almost all the strains that were able to grow at a low temperature (6°C) belonged to cluster W. Most of these strains, including three independently isolated strains, belong to two clonal complexes and are therefore very closely related genetically. These clonal complexes represent strains corresponding to the previously identified species Bacillus weihenstephanensis. Most of the other strains of our collection, including some from the W cluster, are not psychrotrophic. B. weihenstephanensis (cluster W) strains appear to comprise an effectively sexual population, whereas Bacillus thuringiensis (cluster T) and B. cereus (cluster C) have clonal population structures.     

Characteristics of the functioning of the cardio-respiratory system and autonomic regulation in para-athletes with spinal injury

The functional state in wheelchair basketball players with damage to the vertebral column in the area of T ₆-T ₁₀ and paraplegia was studied. The subjects were 26.6 ± 1.7 years old on average (n = 9). Control subjects were disabled persons who led an active life, with a mean age of 44.5 ± 2.6 years (n = 13), athletes with a mean age of 24.6 ± 1.3 years (n = 14), and healthy physically active men with a mean age of 24.9 ± 0.6 years (n = 15). In wheelchair athletes, the body length in the sitting posture, the respiratory volume, and the performance of respiratory tests were increased. These changes in the musculoskeletal apparatus and the systems providing autonomic regulation of motor activity may be regarded as adaptive modifications due to physical training. In the cardiovascular system of para-athletes and its autonomic regulation, attenuation of an increase in the diastolic arterial pressure value induced by injury and an elevation of sensitivity of arterial baroreflex, which had been decreased due to damage to the vertebral column, were observed. These data indicate compensatory processes adjusting the functioning of the cardiovascular system via the mechanisms of baroreflex regulation.     

Population consequences of mass coming of pink salmon Oncorhynchus gorbuscha to the Northeastern Coast of Kamchatka in 2009

During mass spawning of pink salmon Oncorhynchus gorbuscha in two rivers of central Karaginskii Bay (Kamchatka), the average coverage of the spawning hillock did not vary much (0.6?C0.7 m²) even though the abundance of the spawning adults varied greatly. When the abundance of spawning females was less than 1.4 ind. per m² of spawning ground, the spawning hillocks did not interlock. When the abundance of spawning females varied from 1.7 to 1.8 ind./m², the spawning hillocks interlocked, and approximately 20?C25% of them were re-excavated. When the abundance of spawning females was higher than 2.0 ind./m², all the early-deposited spawning hillocks were re-excavated. The re-excavation of the spawning hillocks increased the population mortality also by the mass pre-spawning death of the breeders and spawning outside the spawning grounds. Meanwhile, fish-kill of the eggs in the spawning hillocks was not observed, and the mortality of embryos and larvae did not depend on the spawning intensity. The ratio of live and dead embryos in the spawning hillocks before their hatching was approximately 60%, varying from 44 up to 92% in regard to the environment of the spawning ground (autumn low water, freezing, and siltation). Significant correlation was found between the survivability and grain-size composition of the spawning hillocks when the impact of small-size fraction was a limiting factor.     

Microbiological analysis of the population of extremely haloalkaliphilic sulfur-oxidizing bacteria dominating in lab-scale sulfide-removing bioreactors

Thiopaq biotechnology for partial sulfide oxidation to elemental sulfur is an efficient way to remove H₂S from biogases. However, its application for high-pressure natural gas desulfurization needs upgrading. Particularly, an increase in alkalinity of the scrubbing liquid is required. Therefore, the feasibility of sulfide oxidation into elemental sulfur under oxygen limitation was tested at extremely haloalkaline conditions in lab-scale bioreactors using mix sediments from hypersaline soda lakes as inoculum. The microbiological analysis, both culture dependent and independent, of the successfully operating bioreactors revealed a domination of obligately chemolithoautotrophic and extremely haloalkaliphilic sulfur-oxidizing bacteria belonging to the genus Thioalkalivibrio. Two subgroups were recognized among the isolates. The subgroup enriched from the reactors operating at pH 10 clustered with Thioalkalivibrio jannaschii–Thioalkalivibrio versutus core group of the genus Thioalkalivibrio. Another subgroup, obtained mostly with sulfide as substrate and at lower pH, belonged to the cluster of facultatively alkaliphilic Thioalkalivibrio halophilus. Overall, the results clearly indicate a large potential of the genus Thiolalkalivibrio to efficiently oxidize sulfide at extremely haloalkaline conditions, which makes it suitable for application in the natural gas desulfurization. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users. Nucleotide sequence accession numbers: The GenBank/EMBL accession numbers of the 16S rRNA gene sequence determined in this study are EU709849–EU709878.