Unanticipated coordination of tris buffer to the Radical SAM cluster of the RimO methylthiotransferase

Radical SAM enzymes generally contain a [4Fe–4S]^2+/1+ (RS cluster) cluster bound to the protein via the three cysteines of a canonical motif CxxxCxxC. The non-cysteinyl iron is used to coordinate SAM via its amino-carboxylate moiety. The coordination-induced proximity between the cluster acting as an electron donor and the adenosyl–sulfonium bond of SAM allows for the homolytic cleavage of the latter leading to the formation of the reactive 5′-deoxyadenosyl radical used for substrate activation. Most of the structures of Radical SAM enzymes have been obtained in the presence of SAM, and therefore, little is known about the situation when SAM is not present. In this report, we show that RimO, a methylthiotransferase belonging to the radical SAM superfamily, binds a Tris molecule in the absence of SAM leading to specific spectroscopic signatures both in Mössbauer and pulsed EPR spectroscopies. These data provide a cautionary note for researchers who work with coordinative unsaturated iron sulfur clusters.     

Mycoplasma gallisepticum Inactivated by Targeting the Hydrophobic Domain of the Membrane Preserves Surface Lipoproteins and Induces a Strong Immune Response

An innovative approach for inactivation of Mycoplasma gallisepticum using the hydrophobic photoinduced alkylating probe 1, 5-iodonaphthylazide (INA) is described. Treatment of washed M. gallisepticum mid-exponential culture (0.2 mg cell protein /mL) with INA followed by irradiation with far-ultraviolet light (310–380 nm) completely abolished viability. Transmission electron microscopy showed that the majority of the inactivated M. gallisepticum were comparable in size to intact cells, but that part of the INA-treated M. gallisepticum preparation also contained low density cells and membrane vesicles. Confocal microscopy revealed that untreated M. gallisepticum cells were internalized by chicken red blood cells (c-RBCs), whereas the INA-inactivated cells remained attached to the outer surface of the c-RBCs. INA treatment of M. gallisepticum resulted in a complete inactivation of F0F1 –ATPase and of the L-arginine uptake system, but the cytoplasmatic soluble NADH2 dehydrogenase was only partially affected. Western blot analysis of the lipoprotein fraction showed that the INA-treated M. gallisepticum retained their lipoproteins. Following subcutaneous injection of M. gallisepticum INA-bacterin, 100% and 68.8% of chickens were positive by the rapid serum agglutination test and enzyme-linked immunosorbent assay respectively, 2 weeks post-injection. These data suggest that the photoinducible alkylating agent INA inactivates M. gallisepticum but preserves its surface lipoproteins and thus has the potential to be used as a general approach for the inactivation of mycoplasmas for vaccine development.     

Response of microbial communities to elevated thallium contamination in river sediments

Abstract

The study of microbial communities in river sediments contaminated by thallium (Tl) is necessary to achieve the information for in-situ microbially mediated bioremediation. However, little is known about the microbial community in Tl-contaminated river sediments. In the present study, we characterized the microbial community and their responses to Tl pollution in river sediments from the Tl-mineralized Lanmuchang area, Southwest Guizhou, China. Illumina sequencing of 16S rRNA amplicons revealed that over 40 phyla belong to the domain bacteria. In all samples, Proteobacteria, Cyanobacteria, and Actinobacteria were the most dominant phyla. Based on the UPGMA (Unweighted Pair Group Method with Arithmetic Mean) tree and PCoA (Principal Coordinates Analysis) analysis, microbial composition of each segment was distinct, indicating in-situ geochemical parameters (including Tl, sulfate, TOC, Eh, and pH) had influenced on the microbial communities. Moreover, canonical correspondence analysis (CCA) was employed to further elucidate the impact of geochemical parameters on the distribution of microbial communities in local river sediments. The results indicated that a number of microbial communities including Cyanobacteria, Spirochaete, Hydrogenophaga, and Acinetobacter were positively correlated with total Tl, suggesting potential roles of these microbes to Tl tolerance or to biogeochemical cycling of Tl. Our results suggested a reliable location for the microbial community’s diversity in the presence of high concentrations of Tl and might have a potential association for in-situ bioremediation strategies of Tl-contaminated river. Overall, in situ microbial community could provide a useful tool for monitoring and assessing geo-environmental stressors in Tl-polluted river sediments.     

Design,synthesis, molecular modeling and biological evaluation of novel Benzoxazole-Benzamide conjugates via a 2-Thioacetamido linker as potential anti-proliferative agents,VEGFR-2 inhibitors and apoptotic inducers

A novel series of 2-thioacetamide linked benzoxazole-benzamide conjugates 1–15 was designed as potential inhibitors of the vascular endothelial growth factor receptor-2 (VEGFR-2). The prepared compounds were evaluated for their potential antitumor activity and their corresponding selective cytotoxicity was estimated using normal human fibroblast (WI-38) cells. Compounds 1, 9–12 and 15 showed good selectivity and displayed excellent cytotoxic activity against both HCT-116 and MCF-7 cancer cell lines compared to sorafenib, used as a reference compound. Furthermore, compounds 1 and 11 showed potent VEGFR-2 inhibitory activity. The cell cycle progression assay showed that 1 and 11 induced cell cycle arrest at G2/M phase, with a concomitant increase in the pre-G1 cell population. Further pharmacological studies showed that 1 and 11 induced apoptosis and inhibited the expression of the anti-apoptotic Bcl-2 and Bcl-xL proteins in both cell lines. Therefore, compounds 1 and 11 might serve as promising candidates for future anticancer therapy development.     

New quinoline and isatin derivatives as apoptotic VEGFR-2 inhibitors: design,synthesis, anti-proliferative activity,docking, ADMET,toxicity, and MD simulation studies

New quinoline and isatin derivatives having the main characteristics of VEGFR-2 inhibitors was synthesised. The antiproliferative effects of these compounds were estimated against A549, Caco-2, HepG2, and MDA-MB-231. Compounds 13 and 14 showed comparable activities with doxorubicin against the Caco-2 cells. These compounds strongly inhibited VEGFR-2 kinase activity. The cytotoxic activities were evaluated against Vero cells. Compound 7 showed the highest value of safety and selectivity. Cell migration assay displayed the ability of compound 7 to prevent healing and migration abilities in the cancer cells. Furthermore, compound 7 induced apoptosis in Caco-2 through the expressive down-regulation of the apoptotic genes, Bcl2, Bcl-xl, and Survivin, and the upregulation of the TGF gene. Molecular docking against VEGFR-2 emerged the interactions of the synthesised compounds in a similar way to sorafenib. Additionally, seven molecular dynamics simulations studies were applied and confirmed the stability of compound 13 in the active pocket of VEGFR-2 over 100 ns.     

Nuclear Compartmentalization of Serine Racemase Regulates d-Serine Production: IMPLICATIONS FOR N-METHYL-d-ASPARTATE (NMDA) RECEPTOR ACTIVATION*

d-Serine is a physiological co-agonist that activates N-methyl d-aspartate receptors (NMDARs) and is essential for neurotransmission, synaptic plasticity, and behavior. d-Serine may also trigger NMDAR-mediated neurotoxicity, and its dysregulation may play a role in neurodegeneration. d-Serine is synthesized by the enzyme serine racemase (SR), which directly converts l-serine to d-serine. However, many aspects concerning the regulation of d-serine production under physiological and pathological conditions remain to be elucidated. Here, we investigate possible mechanisms regulating the synthesis of d-serine by SR in paradigms relevant to neurotoxicity. We report that SR undergoes nucleocytoplasmic shuttling and that this process is dysregulated by several insults leading to neuronal death, typically by apoptotic stimuli. Cell death induction promotes nuclear accumulation of SR, in parallel with the nuclear translocation of GAPDH and Siah proteins at an early stage of the cell death process. Mutations in putative SR nuclear export signals (NESs) elicit SR nuclear accumulation and its depletion from the cytosol. Following apoptotic insult, SR associates with nuclear GAPDH along with other nuclear components, and this is accompanied by complete inactivation of the enzyme. As a result, extracellular d-serine concentration is reduced, even though extracellular glutamate concentration increases severalfold. Our observations imply that nuclear translocation of SR provides a fail-safe mechanism to prevent or limit secondary NMDAR-mediated toxicity in nearby synapses.     

Effects of Silver Nanoparticles on Burn Wound Healing in a Mouse Model

Biological Trace Element Research - Healing of injuries caused by exposure to heat has been discussed in many studies, although a few drugs have been shown to produce satisfactory results. In this...     

Interactive effect of nitrogen fertilizer and plant density on photosynthetic and agronomical traits of cotton at different growth stages

Individual effects of application of nitrogen (N) and plant densities (PD) were reported in various studies; however an interactive effect of N and PD in cotton was not studied. To explore the benefits of interactive effects of N fertilizer and PD to increase the quality of cotton. This study was carried out in randomized complete block design (RCBD) with split plot arrangement. In split plot arrangement, main plot was consisted of N application rate and in sub plots different PD were done. There were two nitrogen levels; low N level (F1) 120 kg ha⁻¹ and high N level (F2) 180 kg ha⁻¹ and three planting densities; 8 plants m⁻² as low density (LD), 10 plants m⁻² as medium density (MD) and 12 plants m⁻² as high density (HD). In this study we observed the interactive effect of N application levels and PD on cotton photosynthetic and agronomic traits of various stages of development. Results showed that cotton growth and N contents was varied among treatments on different development stages. Plant biomass production, photosynthetic rate (Pn), intercellular CO₂ (C_i), water use efficiency (WUE) and N contents were unaffected at the seedling stage by N application rate and PD, however, the highest Pn, C_i and N contents was at squaring stage followed by blooming stage. Higher seed cotton yield and lint yield were obtained F1 with HD, and F2 with MD yielded the highest N contents and cotton yield among treatments. We found that the squaring stage was more critical, followed by the blooming stage when considering N rate and PD.     

Less Graft-Versus-Host Disease after Rabbit Antithymocyte Globulin Conditioning in Unrelated Bone Marrow Transplantation for Leukemia and Myelodysplasia: Comparison with Matched Related Bone Marrow Transplantation

One of the major drawbacks for unrelated donor (UD) bone marrow transplantation (BMT) is graft-versus-host disease (GVHD). Despite results from randomized trials, antithymocyte globulin (ATG) is not routinely included for GVHD prophylaxis in UD BMT by many centers. One of ways to demonstrate the usefulness of rabbit ATG in UD BMT is to evaluate how its results approximate to those observed in matched related (MRD) BMT. Therefore, we compared the outcomes between UD BMT with rabbit ATG (Thymoglobulin) for GVHD prophylaxis (n = 25) and MRD BMT (n = 91) for leukemia and myelodysplasia. All but one patient received a myeloablative conditioning regimen. Grades II–IV acute GVHD were similar (39.5% vs. 36%, p = 0.83); however, MRD BMT recipients developed more moderate-severe chronic GVHD (36.5% vs. 8.6%, p = 0.01) and GVHD-related deaths (32.5% vs. 5.6%, p = 0.04). UD BMT independently protected against chronic GVHD (hazard ratio 0.23, p = 0.04). The 6-month transplant-related mortality, 1-year relapse incidence, and 5-year survival rates were similar between patients with non-advanced disease in the MRD and UD BMT groups, 13.8% vs. 16.6% (p = 0.50), 20.8% vs. 16.6% (p = 0.37), and 57% vs. 50% (p = 0.67), respectively. Stable full donor chimerism was equally achieved (71.3% vs. 71.4%, p = 1). Incorporation of rabbit ATG in UD BMT promotes less GVHD, without jeopardizing chimerism evolution, and may attain similar survival outcomes as MRD BMT for leukemia and myelodysplasia especially in patients without advanced disease.     

EEG Spectral Analysis on Muslim Prayers

This study investigated the proposition of relaxation offered by performing the Muslim prayers by measuring the alpha brain activity in the frontal (F3–F4), central (C3–C4), parietal (P3–P4), and occipital (O1–O2) electrode placements using the International 10–20 System. Nine Muslim subjects were asked to perform the four required cycles of movements of Dhuha prayer, and the EEG were subsequently recorded with open eyes under three conditions, namely, resting, performing four cycles of prayer while reciting the specific verses and supplications, and performing four cycles of acted salat condition (prayer movements without any recitations). Analysis of variance (ANOVA) tests revealed that there were no significant difference in the mean alpha relative power (RP_α) between the alpha amplitude in the Dhuha prayer and the acted conditions in all eight electrode positions. However, the mean RP_α showed higher alpha amplitude during the prostration position of the Dhuha prayer and acted condition at the parietal and occipital regions in comparison to the resting condition. Findings were similar to other studies documenting increased alpha amplitude in parietal and occipital regions during meditation and mental concentration. The incidence of increased alpha amplitude suggested parasympathetic activation, thus indicating a state of relaxation. Subsequent studies are needed to delineate the role of mental concentration, and eye focus, on alpha wave amplitude while performing worshipping acts.