NlpC/P60 domain‐containing proteins of Mycobacterium avium subspecies paratuberculosis that differentially bind and hydrolyze peptidoglycan

A subset of proteins containing NlpC/P60 domains are bacterial peptidoglycan hydrolases that cleave noncanonical peptide linkages and contribute to cell wall remodeling as well as cell separation during late stages of division. Some of these proteins have been shown to cleave peptidoglycan in Mycobacterium tuberculosis and play a role in Mycobacterium marinum virulence of zebra fish; however, there are still significant knowledge gaps concerning the molecular function of these proteins in Mycobacterium avium subspecies paratuberculosis (MAP). The MAP genome sequence encodes five NlpC/P60 domain‐containing proteins. We describe atomic resolution crystal structures of two such MAP proteins, MAP_1272c and MAP_1204. These crystal structures, combined with functional assays to measure peptidoglycan cleavage activity, led to the observation that MAP_1272c does not have a functional catalytic core for peptidoglycan hydrolysis. Furthermore, the structure and sequence of MAP_1272c demonstrate that the catalytic residues normally required for hydrolysis are absent, and the protein does not bind peptidoglycan as efficiently as MAP_1204. While the NlpC/P60 catalytic triad is present in MAP_1204, changing the catalytic cysteine‐155 residue to a serine significantly diminished catalytic activity, but did not affect binding to peptidoglycan. Collectively, these findings suggest a broader functional repertoire for NlpC/P60 domain‐containing proteins than simply hydrolases.     

Identification and characterization of clinical Bacillus spp. isolates phenotypically similar to Bacillus anthracis

Bacillus anthracis, the etiological agent of anthrax, is a gram-positive, spore-forming rod, with colonies exhibiting a unique ground-glass appearance, and lacking hemolysis and motility. In addition to these phenotypes, several others traits are characteristic of B. anthracis such as susceptibility to gamma phage, the presence of two virulence plasmids (pX01 and pX02), and specific cell wall and capsular antigens that are commonly detected by direct fluorescent-antibody assays. We report on the identification and characterization of 14 Bacillus megaterium and four Bacillus sp. clinical isolates that are nonhemolytic, nonmotile, and produce a capsule antigenically similar to B. anthracis. This work furthers our understanding of Bacillus diversity and the limitations of the assays and phenotypes that are used to differentiate species in this genus. Further work is necessary to understand whether these strains are opportunistic pathogens or just contaminates.     

Localised invertebrate grazing moderates the effect of warming on competitive fungal interactions

Outcomes of competitive mycelial interactions determine saprotrophic fungal community composition and are regulated by biotic (e.g. invertebrate grazing) and abiotic (e.g. climate) factors. Selective grazing can moderate the effects of elevated temperature on fungal interactions. In natural systems, however, patchy and aggregative distributions of invertebrates exert unequal grazing pressures on competing fungi. We explored whether restricting grazing to the territory of one fungal competitor affected the potential of Oniscus asellus (Isopoda) to control the outcomes of interactions and mediate responses to elevated temperature. Restricted grazing prevented the dominance of any one fungal species in the majority of interactions and, indirectly, altered the influence of warming. The location of grazer restriction was, however, only important during certain interactions. Selective pressures reflected feeding preferences, but grazer location determined the extent of selective grazing pressure exerted. Aggregation of macro-invertebrate grazers appears important in maintaining multi-species assemblages of wood-decomposer fungi in a changing climate.     

Seasonal wetlands on the Lake Wales Ridge, Florida: does a relict seed bank persist despite long term disturbance?

Wetlands maintain biodiversity and provide numerous ecosystem services, so the pressure to perform successful restoration consequently is high. However, restoration projects rarely include an in-depth assessment of wetland potential for recovery, and restoration techniques may not be tailored to site-specific concerns. This study examined the seed bank of disturbed wetlands slotted for hydrologic, but not vegetation, restoration to determine if a seed bank comparable to that of nearby undisturbed wetlands persisted despite long-term anthropogenic disturbance. We compared the aboveground vegetation and seed bank compositions under drained, drawdown, and flooded conditions between undisturbed and historically ditched (“disturbed”) wetlands. Disturbed and undisturbed wetlands shared fewer than 30 % of total aboveground species. While undisturbed wetlands were dominated by graminoids, disturbed wetlands had greater cover of forbs. The seed banks of disturbed wetlands had high species diversity, but their composition was dissimilar to that of nearby undisturbed wetlands. In total, the seed banks of both disturbance histories germinated 56 species; drained conditions had the fewest germinants while flooded conditions had the most. Germinant richness was significantly affected by disturbance, moisture, and their interaction; evenness was significantly affected by moisture, and Shannon diversity by disturbance. Because the seed bank of disturbed wetlands included many fast-growing wetland plants, passive vegetation restoration and active hydrologic restoration may result in wetlands overgrown with weedy species and with fewer conservative wetland plants. An understanding of the capacity for seed banks to re-vegetate wetlands post-restoration and approximate undisturbed wetlands is crucial to the overall success of restoration projects.     

A Pooled Analysis of Body Mass Index and Mortality among African Americans

Pooled analyses among whites and East Asians have demonstrated positive associations between all-cause mortality and body mass index (BMI), but studies of African Americans have yielded less consistent results. We examined the association between BMI and all-cause mortality in a sample of African Americans pooled from seven prospective cohort studies: NIH-AARP, 1995–2009; Adventist Health Study 2, 2002–2008; Black Women''s Health Study, 1995–2009; Cancer Prevention Study II, 1982–2008; Multiethnic Cohort Study, 1993–2007; Prostate, Lung, Colorectal and Ovarian Screening Trial, 1993–2009; Southern Community Cohort Study, 2002–2009. 239,526 African Americans (including 100,175 never smokers without baseline heart disease, stroke, or cancer), age 30–104 (mean 52) and 71% female, were followed up to 26.5 years (mean 11.7). Hazard ratios (HR) and 95% confidence intervals (CI) for mortality were derived from multivariate Cox proportional hazards models. Among healthy, never smokers (11,386 deaths), HRs (CI) for BMI 25–27.4, 27.5–29.9, 30–34.9, 35–39.9, 40–49.9, and 50–60 kg/m² were 1.02 (0.92–1.12), 1.06 (0.95–1.18), 1.32 (1.18–1.47), 1.54 (1.29–1.83), 1.93 (1.46–2.56), and 1.93 (0.80–4.69), respectively among men and 1.06 (0.99–1.15), 1.15 (1.06–1.25), 1.24 (1.15–1.34), 1.58 (1.43–1.74), 1.80 (1.60–2.02), and 2.31 (1.74–3.07) respectively among women (reference category 22.5–24.9). HRs were highest among those with the highest educational attainment, longest follow-up, and for cardiovascular disease mortality. Obesity was associated with a higher risk of mortality in African Americans, similar to that observed in pooled analyses of whites and East Asians. This study provides compelling evidence to support public health efforts to prevent excess weight gain and obesity in African Americans.     

A comparison of stereology, structural rigidity and a novel 3D failure surface analysis method in the assessment of torsional strength and stiffness in a mouse tibia fracture model

In attempting to develop non-invasive image based measures for the determination of the biomechanical integrity of healing fractures, traditional μCT based measurements have been limited. This study presents the development and evaluation of a tool for assessment of fracture callus mechanical properties through determination of the geometric characteristics of the fracture callus, specifically along the surface of failure identified during destructive mechanical testing. Fractures were created in tibias of ten male mice and subjected to μCT imaging and biomechanical torsion testing. Failure surface analysis, along with previously described image based measures was calculated using the μCT image data, and correlated with mechanical strength and stiffness. Three-dimensional measures along the surface of failure, specifically the surface area and torsional rigidity of bone, were shown to be significantly correlating with mechanical strength and stiffness. It was also shown that surface area of bone along the failure surface exhibits stronger correlations with both strength and stiffness than measures of average and minimum torsional rigidity of the entire callus. Failure surfaces observed in this study were generally oriented at 45° to the long axis of the bone, and were not contained exclusively within the callus. This work represents a proof of concept study, and shows the potential utility of failure surface analysis in the assessment of fracture callus stability.     

Nitrogen mineralization in high-elevation forests of the appalachians. II. Patterns with stand development in fir waves

Wave-like patterns of mortality and regeneration of balsam fir (Abies balsamea {L.} Mill.) and Fraser fir (A. fraseri {Pursh.} Poir.) forests at high elevations in the Appalachian Mountains offer a unique opportunity to study the effects of stand development on nitrogen cycling. We sampled two fir waves, one with Fraser fir on Mt. LeConte in the Great Smoky Mountains National Park, and one with balsam fir on Whiteface Mountain in New York. Net nitrogen mineralization for 3 summer months at Mt. LeConte was high in the dead fir zone (47 kg-N/ha), lower in the regeneration and juvenile zones (24 and 21 kg-N/ha), and highest in the mature zone (61 kg-N/ha). This sampling period probably accounted for about 60% of the annual total. The pattern was similar in the balsam fir wave on Whiteface Mountain, with N mineralization rates of 39 and 33 kg-N/ha over 2 months for the regenerating and juvenile zones, and 43 and 54 kg-N/ha for the mature and dead zones. Throughfall nitrogen followed a fairly similar pattern, with rates ranging from 4.5 to 10 kg-N/ha for 2 or 3 months across all zones at both sites. Tension-free lysimeters indicated very little leaching of nitrogen below 30 cm depth (the maximum was 2.6 kg-N/ha), but these estimates may be low. We conclude that nitrogen mineralization is high at all stages of stand development, perhaps exceeding the uptake capacity of the trees. Rates of nitrogen leaching may be high in these ecosystems and should receive further attention. Request for offprints     

Potentials for win-win alliances among animal agriculture and forest products industries: Application of the principles of industrial ecology and sustainable development

Commercial forests in many parts of the world are deficient in nitrogen and phosphorus. These nutrient-deficient forests often exist in close proximity to large animal feeding operations, meat processing and other food, textile, or other biomass-processing plants, and municipal waste treatment facilities. Many of these facilities produce large surpluses of nitrogen, phosphorus, and organic matter as gaseous ammonia, urea, uric acid, phosphorus compounds, bacterial sludges, and partially treated municipal wastewaters. These co-existing and substantial nutrient deficiencies and surpluses offer ready-made opportunities for discovery, demonstration, and commercial development of science-based, technology-facilitated, environmentally sound, economically viable, and socially acceptable “win-win alliances” among these major industries based on the principles of industrial ecology and sustainable development. The major challenge is to discover practical means to capture the surplus nutrients and put them to work in forest stands from which value-added products can be produced and sold at a profit.