The glucose metabolite methylglyoxal inhibits expression of the glucose transporter genes by inactivating the cell surface glucose sensors Rgt2 and Snf3 in yeast

Methylglyoxal (MG) is a cytotoxic by-product of glycolysis. MG has inhibitory effect on the growth of cells ranging from microorganisms to higher eukaryotes, but its molecular targets are largely unknown. The yeast cell-surface glucose sensors Rgt2 and Snf3 function as glucose receptors that sense extracellular glucose and generate a signal for induction of expression of genes encoding glucose transporters (HXTs). Here we provide evidence that these glucose sensors are primary targets of MG in yeast. MG inhibits the growth of glucose-fermenting yeast cells by inducing endocytosis and degradation of the glucose sensors. However, the glucose sensors with mutations at their putative ubiquitin-acceptor lysine residues are resistant to MG-induced degradation. These results suggest that the glucose sensors are inactivated through ubiquitin-mediated endocytosis and degraded in the presence of MG. In addition, the inhibitory effect of MG on the glucose sensors is greatly enhanced in cells lacking Glo1, a key component of the MG detoxification system. Thus the stability of these glucose sensors seems to be critically regulated by intracellular MG levels. Taken together, these findings suggest that MG attenuates glycolysis by promoting degradation of the cell-surface glucose sensors and thus identify MG as a potential glycolytic inhibitor.     

Development of sensitive benzofurazan‐based spectrometric methods for analysis of spectinomycin in vials and human biological samples

Spectinomycin hydrochloride (SPEC) is an aminoglycoside antibiotic that has a broad spectrum against several bacterial strains of humans and veterinary infections. However, SPEC lacks a fluorophore or chromophore and this lack makes its analysis a challenge. Our study provides a simple, sensitive and low‐cost spectrofluorimetric/spectrophotometric method based on the reaction between secondary amine groups and a benzofurazan reagent using borate buffer (pH 9.2). The yielding compound was measured fluorimetrically at 530 nm (excitation at 460 nm) with colorimetry at 410 nm. The calibration curves ranged from 30 to 400 ng ml^?1 and from 0.2 to 6 μg ml^?1 for spectrofluorimetric and spectrophotometric analyses, respectively. The limits of detection were calculated to be 4.15 ng ml^?1 and 0.05 μg ml^?1 for spectrofluorimetric and spectrophotometric processes, respectively. The ultra‐affectability and high selectivity of the proposed method permitted analysis of SPEC in the dosage form and in human plasma samples, with good recoveries of about 101.19 and 97.11%, respectively, without any matrix interference. The proposed strategy was validated using International Conference on Harmonization standards and validated bioanalytically using USFDA recommendations.     

Phragmites australis — a helophytic grass — can establish successful partnership with phenol-degrading bacteria in a floating treatment wetland

Helophytic plants contribute significantly in phytoremediation of a variety of pollutants due to their physiological or biochemical mechanisms. Phenol, which is reported to have negative/deleterious effects on plant metabolism at concentrations higher than 500 mg/L, remains hard to be removed from the environmental compartments using conventional phytoremediation procedures. The present study aims to investigate the feasibility of using P. australis (a helophytic grass) in combination with three bacterial strains namely Acinetobacter lwofii ACRH76, Bacillus cereus LORH97, and Pseudomonas sp. LCRH90, in a floating treatment wetland (FTW) for the removal of phenol from contaminated water. The strains were screened based on their phenol degrading and plant growth promoting activities. We found that inoculated bacteria were able to colonize in the roots and shoots of P. australis, suggesting their potential role in the successful removal of phenol from the contaminated water. Pseudomonas sp. LCRH90 dominated the bacterial community structure followed by A. lowfii ACRH76 and B. cereus LORH97. The removal rate was significantly high when compared with the individual partners, i.e., plants and bacteria separately. The plant biomass, which was drastically reduced in the presence of phenol, recovered significantly with the inoculation of bacterial consortia. Likewise, highest reduction in chemical oxygen demand (COD), biochemical oxygen demand (BOD), and total organic carbon (TOC) is achieved when both plants and bacteria were employed. The study, therefore, suggests that P. australis in combination with efficient bacteria can be a suitable choice to FTWs for phenol-degradation in water.     

Size and time-dependent induction of proinflammatory cytokines expression in brains of mice treated with gold nanoparticles

Gold nanoparticles (GNPs) are among the ideal nano-sized materials for medical applications such as imaging and drug delivery. Considering the significance of recent reports on acute phase induction of inflammatory mediators by GNPs, we studied the effect of GNPs on proinflammatory cytokines gene expression in mouse brain. Group 1 served as control whereas groups 2–4 were given only one intraperitoneal dose of 5, 20 and 50?nm GNPs, respectively and sacrificed after 24?h. The animals in groups 5–7 also received the same treatment but sacrificed after 7?days. Groups 8–10 received two injections of GNPs (5, 20 and 50?nm, respectively), first at the beginning of study and second on day 6, and sacrificed on day 7. Total RNA was extracted from the cerebral tissue and analyzed for the gene expressions of IL-1β, IL-6 and TNF-α. A single injection of 5?nm diameter GNPs significantly increased the mRNA expression of IL-1β and IL-6 in mouse brain on day 7, which was not augmented by the second dose of the same GNPs. Larger size GNPs (20?nm and 50?nm) did not cause any significant change in the expression of proinflammatory cytokines in mouse brain. In conclusion, systemic administration of small sized GNPs (5?nm) induced a proinflammatory cascade in mouse brain indicating a crucial role of GNPs size on immune response. It is important to use the right sized GNPs in order to avoid an acute phase inflammatory response that could be cytotoxic or interfere with the bioavailability of nanomaterials.     

A comparative study of bacterial diversity based on culturable and culture-independent techniques in the rhizosphere of maize (Zea mays L.)

ObjectiveMaize is an important crop for fodder, food and feed industry. The present study explores the plant-microbe interactions as alternative eco-friendly sustainable strategies to enhance the crop yield.MethodologyBacterial diversity was studied in the rhizosphere of maize by culture-dependent and culture-independent techniques by soil sampling, extraction of DNA, amplification of gene of interest, cloning of desired fragment and library construction.ResultsCulturable bacteria were identified as Achromobacter, Agrobacterium, Azospirillum, Bacillus, Brevibacillus, Bosea, Enterobacter, Microbacterium, Pseudomonas, Rhodococcus, Stenotrophomonas and Xanthomonas genera. For culture-independent approach, clone library of 16S ribosomal RNA gene was assembled and 100 randomly selected clones were sequenced. Majority of the sequences were related to Firmicutes (17%), Acidobacteria (16%), Actinobacteria (17%), Alpha-Proteobacteria (7%), Delta-proteobacteria (4.2%) and Gemmatimonadetes (4.2%) However, some of the sequences (30%) were novel that showed no homologies to phyla of cultured bacteria in the database. Diversity of diazotrophic bacteria in the rhizosphere investigated by analysis of PCR-amplified nifH gene sequence that revealed abundance of sequences belonging to genera Azoarcus (25%), Aeromonas (10%), Pseudomonas (10%). The diazotrophic genera Azotobacter, Agrobacterium and Zoogloea related nifH sequences were also detected but no sequence related to Azospirillum was found showing biasness of the growth medium rather than relative abundance of diazotrophs in the rhizosphere.ConclusionThe study provides a foundation for future research on focussed isolation of the Azoarcus and other diazotrophs found in higher abundance in the rhizosphere.     

Physical and chemical mutagens improved Sporotrichum thermophile,strain ST20 for enhanced Phytase activity

ObjectivePhosphorous is an essential micronutrient of plants and involved in critical biological functions. In nature, phosphorous is mostly present in immobilized inorganic mineral and in the fixed organic form including phytic acid and phosphoesteric compounds. However, the bioavailability of bound phosphorous could be enhanced by the use of phosphate solubilizing microorganisms such as bacteria and fungi. The phytases are widespread in an environment and have been isolated from different sources comprising bacteria and fungi.MethodologyIn current studies, we show the successful use of gamma rays and EMS (Ethyl Methane Sulphonate) mutagenesis for enhanced activity of phytases in a fungal strain Sporotrichum thermophile.ResultsWe report an improved strain ST2 that could produce a clear halo zone around the colony, up to 24 mm. The maximum enzymatic activity was found of 382 U/mL on pH 5.5. However, the phytase activity was improved to 387 U/ml at 45 °C. We also report that the mutants produced through EMS showed the greater potential for phytase production.ConclusionThe current study highlights the potential of EMS mutagenesis for strain improvement over physical mutagens.     

High intrinsic seed Zn concentration improves abiotic stress tolerance in wheat

Plant and Soil - Abiotic stresses are threatening wheat productivity across the globe, which is often associated with nutrient deficiencies. Zinc (Zn) is involved in many physiological processes of...     

Reasons for relinquishment of companion animals in U.S. animal shelters: selected health and personal issues

In personal interviews, people surrendering their dogs and cats to 12animal shelters in 4 regions of the country discussed their reasons for relinquishing their companion animals and answered questions about their own characteristics and those of their pet. The interviews identified 71 reasons for relinquishment. Personal issues lead the class of reasons for relinquishment of cats and ranked 3rd among those given for relinquishment of dogs. The top 3 health and personal issues cited for giving up cats were allergies of a family member to cats, owner's personal problems, and anew baby. For dogs, the top 3 reasons cited were lack of time for the dog, owner's personal problems, and allergies. Analysis of these health and personal issues suggests that education and counseling before and after acquisition of a pet, as well as the availability of temporary accommodations for pets during times of personal crisis, may reduce relinquishments.