Phosphoglycerate mutase from Trypanosoma brucei is hyperactivated by cobalt in vitro, but not in vivo

Production of ATP by the glycolytic pathway in the mammalian pathogenic stage of protists from the genus Trypanosoma is required for the survival of the parasites. Cofactor-independent phosphoglycerate mutase (iPGAM) is particularly attractive as a drug target because it shows no similarity to the corresponding enzyme in humans, and has also been genetically validated as a target by RNAi experiments. It has previously been shown that trypanosomatid iPGAMs require Co(2+) to reach maximal activity, but the biologically relevant metal has remained unclear. In this paper the metal content in the cytosol of procyclic and bloodstream-form T. brucei (analysed by inductively coupled plasma-optical emission spectroscopy) shows that Mg(2+), Zn(2+) and Fe(2+) were the most abundant, whereas Co(2+) was below the limit of detection (<0.035 μM). The low concentration indicates that Co(2+) is unlikely to be the biologically relevant metal, but that instead, Mg(2+) and/or Zn(2+) may assume this role. Results from metal analysis of purified Leishmania mexicana iPGAM by inductively coupled plasma-mass spectrometry also show high concentrations of Mg(2+) and Zn(2+), and are consistent with this proposal. Our data suggest that in vivo cellular conditions lacking Co(2+) are unable to support the maximal activity of iPGAM, but instead maintain its activity at a relatively low level by using Mg(2+) and/or Zn(2+). The physiological significance of these observations is being pursued by structural, biochemical and biophysical studies.     

Heritability and coefficient of genetic variation analyses of phenotypic traits provide strong basis for high-resolution QTL mapping in the Collaborative Cross mouse genetic reference population

Most biological traits of human importance are complex in nature; their manifestation controlled by the cumulative effect of many genetic factors interacting with one another and with the individual’s life history. Because of this, mouse genetic reference populations (GRPs) consisting of collections of inbred lines or recombinant inbred lines (RIL) derived from crosses between inbred lines are of particular value in analysis of complex traits, since massive amounts of data can be accumulated on the individual lines. However, existing mouse GRPs are derived from inbred lines that share a common history, resulting in limited genetic diversity, and reduced mapping precision due to long-range gametic disequilibrium. To overcome these limitations, the Collaborative Cross (CC) a genetically highly diverse collection of mouse RIL was established. The CC, now in advanced stages of development, will eventually consist of about 500 RIL derived from reciprocal crosses of eight divergent founder strains of mice, including three wild subspecies. Previous studies have shown that the CC indeed contains enormous diversity at the DNA level, that founder haplotypes are inherited in expected frequency, and that long-range gametic disequilibrium is not present. We here present data, primarily from our own laboratory, documenting extensive genetic variation among CC lines as expressed in broad-sense heritability (H²) and by the well-known “coefficient of genetic variation,” demonstrating the ability of the CC resource to provide unprecedented mapping precision leading to identification of strong candidate genes.     

Marine bacteria and omic approaches: A novel and potential repository for bioremediation assessment

Marine environments accommodating diverse assortments of life constitute a great pool of differentiated natural resources. The cumulative need to remedy unpropitious effects of anthropogenic activities on estuaries and coastal marine ecosystems has propelled the development of effective bioremediation strategies. Marine bacteria producing biosurfactants are promising agents for bio-remediating oil pollution in marine environments, making them prospective candidates for enhancing oil recovery. Molecular omics technologies are considered an emerging field of research in ecological and diversity assessment owing to their utility in environmental surveillance and bioremediation of polluted sites. A thorough literature review was undertaken to understand the applicability of different omic techniques used for bioremediation assessment using marine bacteria. This review further establishes that for bioremediation of environmental pollutants (i.e. heavy metals, hydrocarbons, xenobiotic and numerous recalcitrant compounds), organisms isolated from marine environments can be better used for their removal. The literature survey shows that omics approaches can provide exemplary knowledge about microbial communities and their role in the bioremediation of environmental pollutants. This review centres on applications of marine bacteria in enhanced bioremediation, using the omics approaches that can be a vital biological contrivance in environmental monitoring to tackle environmental degradation. The paper aims to identify the gaps in investigations involving marine bacteria to help researchers, ecologists and decision-makers to develop a holistic understanding regarding their utility in bioremediation assessment.     

Epstein-Barr virus-transformed lymphoblastoid cell lines as antigen-presenting cells and "augmenting" cells for human CMV-specific Th clones

The ability of Epstein-Barr virus-transformed lymphoblastoid cell lines (LCL) to present human cytomegalovirus (HCMV) antigen to a panel of HCMV-specific T helper (Th) clones was evaluated. Among the seven Th clones studied, only one clone (SP-CN/T3-16) proliferated well to HCMV presented by both autologous mononuclear cells (MNC) and LCL, and one clone (SP-CN/T3-9) proliferated significantly better to HCMV presented by autologous LCL than by autologous MNC. The majority of the HCMV-specific Th clones tested (five out of seven) responded much better to HCMV presented by MNC than to HCMV presented by LCL. The mechanism(s) responsible for the inefficiency of LCL to present HCMV to certain clones was studied. Our results suggested that the defect of LCL is not due to insufficient interleukin 1 production, insufficient MHC class II molecule expression, nor an inhibitory mechanism or factor. In this report, we also demonstrate that by adding a minimum amount of LCL along with MNC as antigen-presenting cells (APC), one can restimulate and expand Th clones much more efficiently than by using MNC alone as APC.     

Removing biofilms from microstructured titanium ex vivo: a novel approach using atmospheric plasma technology

The removal of biofilms from microstructured titanium used for dental implants is a still unresolved challenge. This experimental study investigated disinfection and removal of in situ formed biofilms from microstructured titanium using cold atmospheric plasma in combination with air/water spray. Titanium discs (roughness (Ra): 1.96 µm) were exposed to human oral cavities for 24 and 72 hours (n = 149 each) to produce biofilms. Biofilm thickness was determined using confocal laser scanning microscopy (n = 5 each). Plasma treatment of biofilms was carried out ex vivo using a microwave-driven pulsed plasma source working at temperatures from 39 to 43°C. Following plasma treatment, one group was air/water spray treated before re-treatment by second plasma pulses. Vital microorganisms on the titanium surfaces were identified by contact culture (Rodac agar plates). Biofilm presence and bacterial viability were quantified by fluorescence microscopy. Morphology of titanium surfaces and attached biofilms was visualized by scanning electron microscopy (SEM). Total protein amounts of biofilms were colorimetrically quantified. Untreated and air/water treated biofilms served as controls. Cold plasma treatment of native biofilms with a mean thickness of 19 µm (24 h) to 91 µm (72 h) covering the microstructure of the titanium surface caused inactivation of biofilm bacteria and significant reduction of protein amounts. Total removal of biofilms, however, required additional application of air/water spray, and a second series of plasma treatment. Importantly, the microstructure of the titanium discs was not altered by plasma treatment. The combination of atmospheric plasma and non-abrasive air/water spray is applicable for complete elimination of oral biofilms from microstructured titanium used for dental implants and may enable new routes for the therapy of periimplant disease.     

The immunolocalization of small nuclear ribonucleoprotein particles in testicular cells during the cycle of the seminiferous epithelium of the adult rat

The objective of this study was to determine the cellular and subcellular distribution of small nuclear ribonucleoprotein particles (snRNPs) in the adult rat testis in relation to the different cell types at the various stages of the cycle of the seminiferous epithelium. The distribution of snRNPs in the nucleus and cytoplasm of germ cells was quantitated in an attempt to correlate RNA processing with morphological and functional changes occurring during the development of these cells. Light-microscopic immunoperoxidase staining of rat testes with polyclonal anti-Sm and monoclonal anti-Y12 antibodies localized spliceosome snRNPs in the nuclei and cytoplasm of germ cells up to step 10 spermatids. Nuclear staining was intense in Sertoli cells, spermatogonia, spermatocytes, and in the early steps of round spermatid development. Although comparatively weaker, cytoplasmic staining for snRNPs was strongest in mid and late pachytene spermatocytes and early round spermatids. Quantitative electron-microscopic immunogold labeling of Lowicryl embedded testicular sections confirmed the light-microscopic observations but additionally showed that the snRNP content peaked in the cytoplasm of midpachytene spermatocytes and in the nuclei of late pachytene spermatocytes. The immunogold label tended to aggregate into distinct loci over the nuclear chromatin. The chromatoid body of spermatids and spermatocytes and the finely granular material in the interstices of mitochondrial aggregates of spermatocytes were found to be additional sites of snRNP localization and were intensely labeled. This colocalization suggests that these dense cytoplasmic structures may be functionally related. Anti-U1 snRNP antibodies applied to frozen sections showed the same LM localization pattern as spliceosome snRNPs. Anti-U3 snRNP antibodies applied to frozen sections stained nucleoli of germ cells where pre-rRNA is spliced.     

Balance between microbial calcification and metazoan bioerosion in modern stromatolitic oncolites

Stromatolites date back some 3.5 billion years and constitute the most common and conspicuous fossils through the Proterozoic. These organosedimentary structures decreased dramatically in diversity and abundance by the late Neoproterozoic, a phenomenon often ascribed to destructive grazing by newly evolved metazoans. We investigated the concurrent processes of microbial calcification and metazoan bioerosion in one of the few locations (Rio Mesquites, Cuatro Ciénegas, Coahuila, Mexico) where living freshwater stromatolites, formed by cyanobacteria and diatoms, coexist with significant populations of metazoan grazers. We used microsensor chemical profiling and monitoring of bulk water Ca²⁺ concentrations to determine calcification rates and their dependence on microbial metabolism. The bioerosive impact resulting from grazing by endemic hydrobiid gastropods was assessed by gravimetric quantification of carbonaceous faecal pellet production. Calcification was clearly light‐dependent, reaching maximal rates (saturation) at low incident light intensity, and was surprisingly efficient, with O₂/Ca²⁺ exchange ratios well above unity, and with absolute rates similar to those found in corals. However, the erosive action of grazing snails removed most of these carbonate inputs from the oncolites. Thus, a precarious balance between constructive and destructive geobiological processes was at play in the system. The fact that accretion barely exceeded bioerosion in an environment highly conducive to calcification supports the potential impact of faunal grazing as causal agent in the demise of stromatolites in the late Proterozoic. Our findings indicate that a search for fossil evidence of bioerosive grazing in the form of carbonaceous faecal pellets associated with fossil stromatolites may provide a means to test that hypothesis directly.     

Weight change during pharmacological blockade of interleukin-6 or tumor necrosis factor-α in patients with inflammatory rheumatic disorders: A 16-week comparative study

BackgroundInterleukin (IL)-6 ?/? mice develop spontaneous mature onset obesity, while the influence of the pharmacological blockade of IL-6 on body weight in humans has not been previously reported. The aim of the present study was to observe weight change in patients treated with tocilizumab (TCZ).MethodsTwenty-one consecutive patients who started new treatment with TCZ were enrolled in the study. Sixteen consecutive patients who started treatment with infliximab (IFX) formed the control group. Height and weight of all patients were registered and Body Mass Index (BMI) calculated before the first treatment and at week 16. The Mann–Whitney or paired Wilcoxon test were used for comparisons between or within groups, respectively.ResultsThe study demonstrated that treatment with TCZ was accompanied with significant weight gain and BMI increase (p = 0.04), while IFX treatment did not result in any significant weight change during the 16-week period.ConclusionsWeight gain can be seen in some patients during the pharmacological blockade of IL-6. The phenomenon and metabolic pathways involved should be further investigated.