Rpf proteins are the factors of reactivation of the dormant forms of actinobacteria

As the response to unfavorable growth conditions, nonsporulating mycobacteria transform into the dormant state with the concomitant formation of the specialized dormant forms characterized by low metabolic activity and resistance to antibiotics. Such dormant cells can be reactivated under the influence of several factors including proteins of Rpf (Resuscitation promoting factor) family, which possess peptidoglycan hydrolase activity and were considered to belong to the group of the autocrine growth factors of the bacteria. Remarkable interest toward Rpf family is determined by its par-ticipation in resuscitation of the dormant forms of Mycobacterium tuberculosis, what in turn is the key element in resuscitation of the latent tuberculosis – an infectious disease that affects one third of the World’s population. Experiments with Rpf mutant forms and with strains deleted in these proteins revealed a relationship between the enzymatic activity of this protein and its ability to resuscitate mycobacteria both in vitro and in vivo. This review discusses possible mechanisms of Rpf action including those related to possible participation of the products of mycobacterial Rpf-mediated cell wall hydrolysis (muropeptides) as signaling molecules. The unique ability of Rpf proteins to resuscitate the dormant forms of mycobacteria and to stimulate their proliferation would allow these proteins to occupy their niche in medicine–in diagnostics and in creation of antituberculosis subunit vaccines.     

Scale- and taxon-dependent patterns of plant diversity in steppes of Khakassia,South Siberia (Russia)

The drivers of plant richness at fine spatial scales in steppe ecosystems are still not sufficiently understood. Our main research questions were: (i) How rich in plant species are the natural steppes of Southern Siberia compared to natural and semi-natural grasslands in other regions of the Palaearctic? (ii) What are the main environmental drivers of the diversity patterns in these steppes? (iii) What are the diversity–environment relationships and do they vary between spatial scales and among different taxonomic groups? We sampled the steppe vegetation (vascular plants, bryophytes and lichens) in Khakassia (Russia) with 39 nested-plot series (0.0001–100-m² plot size) and 54 additional 10-m² quadrats across the regional range of steppe types and measured various environmental variables. We measured β-diversity using z-values of power-law species–area relationships. GLM analyses were performed to assess the importance of environmental variables as predictors of species richness and z-value. Khakassian steppes showed both high α- and β-diversity. We found significant scale dependence for the z-values, which had their highest values at small spatial scales and then decreased exponentially. Total species richness was controlled predominantly by heat load index, mean annual precipitation, humus content and soil skeleton content. The positive role of soil pH was evident only for vascular plant species richness. Similar to other studies, we found that the importance of environmental factors strongly differed among taxonomic groups and across spatial scales, thus highlighting the need to study more than one taxon and more than one plot size to get a reliable picture.     

Fodder resources and feeding of semifree bison (<Emphasis Type="Italic">Bison bison</Emphasis>) on a steppe pasture in the Western Manych valley

In 2012–2013, the species composition, aboveground phytomass, and nutrition of freely ranging bison were investigated on a natural steppe pasture of the Western Manych valley. The aboveground mass of plants on the investigated pasture varied from 16.1 to 19.4 c/ha. The composition of the plants consumed by bison and their share in the bison’s diet were determined by microscopy of feces. In the snowless season, graminoids was the basis of the bison’s diet (79–87%). The use of forbs varied from 4.7 to 17.5%. Sedges were present in the forage in summer; they amounted to 6.5–15.4%. The quantitative assessment of the food selectivity was determined using the ratio of the share of a plant species in the diet to its portion in the plant community of the pasture. The most selected were Festuca valesiaca (the selectivity is 12.0), Agropyron sp. sp. (1.7), Polygonum patulum (1.2), and Carex sp. sp. (1.1). In other forage plants (Poa bulbojsa, Tanacetum achileifolium, and other forbs), the selectivity was 0.01–0.3. The nutritional value of fodder vegetation and food indexes (daily intake, digestibility of forage, consumption of metabolizable energy) were estimated. The digestibility was determined by the ratio of inert (undigestible) components (silicon, lignin) in the diet to their amount in the feces. The daily intake of forage was calculated based on the mass of released feces and digestibility of forage. In the snowless season, bison consumed from 6.3–7.0 to 9.2–9.9 kg/ind. (dry weight) per day. The maximum consumption (9.9 kg/ind.) was observed in summer. The digestibility of forage changed from 45 to 53% (the average is 50%). The consumption of metabolizable energy in different seasons varied from 0.60 to 0.91 MJ/kg^0.75/day, which corresponded to maintenance energy.     

Mechanisms of intracellular trigger signal transmission in muscles: Strategy of rearrangements in evolution of the contractile function

The paper describes our concept about the existence of a certain strategy of rearrangements of ionic mechanisms of he intracellular trigger signal transmission in muscles during their contractile function evolution. It is shown that the rearrangements of muscles to accelerate the single (discrete) contraction cycle is accompanied by a change of mechanisms of external stimulus transduction into an intracellular trigger signal: direct activation of intracellular effectors by extracellular Ca²⁺ is replaced by indirect mechanisms of Ca²⁺-, then Ca²⁺- and Na⁺-induced, and in skeletal muscle fibers of vertebrates (SMFV) of Na⁺-induced Ca²⁺ release from the intracellular depot, sarcoplasmic reticulum. These rearrangements promoted an intensification of the Ca²⁺ intracellular mobilization to provide for the most complete pulse control of SMFV phasic contractions by the CNS and their protection from undesirable peripheral influences.     

Etiological diagnosis of suppurative bacterial meningitis today

As the result of laboratory examination of 2165 patients with virulent bacterial meningitides, including cases of meningococcal infection, the etiological diagnosis was confirmed in 1407 patients (65.0%), the number of cases confirmed by the laboratory examination being significantly greater among adults than among children: 67.5 +/- 1.37% and 63.1 +/- 1.53%, respectively, (t = 2.1). Meningococcal infection was confirmed in 1111 (70.6%) out of 1572 patients under examination. Of the patients with purulent meningitides, pneumococcal etiology was determined in 27.4%, type b of Haemophilus influenzae in 13.5%, other infective agents in 10.0%. The comparison of the results obtained in the examination, carried out by different methods, of 946 children and 770 adults with meningococcal infection revealed a considerable difference in the number of positive results yielded by the bacteriological method and countercurrent immunoelectrophoresis (CIE). Among adults meningococci were isolated twice as frequently (41.1 +/- 2.5% - 19.4 +/- 1.6%), and the results yielded by CIE were predominantly positive (55.1 +/- 2.5% and 40.1 +/- 2.5%). CIE and the immunoenzyme assay were shown to have advantages in the diagnosis of the disease. Under the conditions of intensive antibiotic therapy the methods based on the detection of specific antigens in body fluids can greatly assist in diagnosis.     

Muralytic activity of Micrococcus luteus Rpf and its relationship to physiological activity in promoting bacterial growth and resuscitation

The culturability of several actinobacteria is controlled by resuscitation-promoting factors (Rpfs). These are proteins containing a c. 70-residue domain that adopts a lysozyme-like fold. The invariant catalytic glutamate residue found in lysozyme and various bacterial lytic transglycosylases is also conserved in the Rpf proteins. Rpf from Micrococcus luteus, the founder member of this protein family, is indeed a muralytic enzyme, as revealed by its activity in zymograms containing M. luteus cell walls and its ability to (i) cause lysis of Escherichia coli when expressed and secreted into the periplasm; (ii) release fluorescent material from fluorescamine-labelled cell walls of M. luteus; and (iii) hydrolyse the artificial lysozyme substrate, 4-methylumbelliferyl-beta-D-N,N',N'-triacetylchitotrioside. Rpf activity was reduced but not completely abolished when the invariant glutamate residue was altered. Moreover, none of the other acidic residues in the Rpf domain was absolutely required for muralytic activity. Replacement of one or both of the cysteine residues that probably form a disulphide bridge within Rpf impaired but did not completely abolish muralytic activity. The muralytic activities of the Rpf mutants were correlated with their abilities to stimulate bacterial culturability and resuscitation, consistent with the view that the biological activity of Rpf results directly or indirectly from its ability to cleave bonds in bacterial peptidoglycan.     

Latitudinal patterns of magnitude and interannual variability in net ecosystem exchange regulated by biological and environmental variables

Over the last two and half decades, strong evidence showed that the terrestrial ecosystems are acting as a net sink for atmospheric carbon. However the spatial and temporal patterns of variation in the sink are not well known. In this study, we examined latitudinal patterns of interannual variability (IAV) in net ecosystem exchange (NEE) of CO₂ based on 163 site-years of eddy covariance data, from 39 northern-hemisphere research sites located at latitudes ranging from ∼29°N to ∼64°N. We computed the standard deviation of annual NEE integrals at individual sites to represent absolute interannual variability (AIAV), and the corresponding coefficient of variation as a measure of relative interannual variability (RIAV). Our results showed decreased trends of annual NEE with increasing latitude for both deciduous broadleaf forests and evergreen needleleaf forests. Gross primary production (GPP) explained a significant proportion of the spatial variation of NEE across evergreen needleleaf forests, whereas, across deciduous broadleaf forests, it is ecosystem respiration (Re). In addition, AIAV in GPP and Re increased significantly with latitude in deciduous broadleaf forests, but AIAV in GPP decreased significantly with latitude in evergreen needleleaf forests. Furthermore, RIAV in NEE, GPP, and Re appeared to increase significantly with latitude in deciduous broadleaf forests, but not in evergreen needleleaf forests. Correlation analyses showed air temperature was the primary environmental factor that determined RIAV of NEE in deciduous broadleaf forest across the North American sites, and none of the chosen climatic factors could explain RIAV of NEE in evergreen needleleaf forests. Mean annual NEE significantly increased with latitude in grasslands. Precipitation was dominant environmental factor for the spatial variation of magnitude and IAV in GPP and Re in grasslands.