The proton-activated G protein coupled receptor OGR1 acutely regulates the activity of epithelial proton transport proteins

The Ovarian cancer G protein-coupled Receptor 1 (OGR1; GPR68) is proton-sensitive in the pH range of 6.8 - 7.8. However, its physiological function is not defined to date. OGR1 signals via inositol trisphosphate and intracellular calcium, albeit downstream events are unclear. To elucidate OGR1 function further, we transfected HEK293 cells with active OGR1 receptor or a mutant lacking 5 histidine residues (H5Phe-OGR1). An acute switch of extracellular pH from 8 to 7.1 (10 nmol/l vs 90 nmol/l protons) stimulated NHE and H(+)-ATPase activity in OGR1-transfected cells, but not in H5Phe-OGR1-transfected cells. ZnCl(2) and CuCl(2) that both inhibit OGR1 reduced the stimulatory effect. The activity was blocked by chelerythrine, whereas the ERK1/2 inhibitor PD 098059 had no inhibitory effect. OGR1 activation increased intracellular calcium in transfected HEK293 cells. We next isolated proximal tubules from kidneys of wild-type and OGR1-deficient mice and measured the effect of extracellular pH on NHE activity in vitro. Deletion of OGR1 affected the pH-dependent proton extrusion, however, in the opposite direction as expected from cell culture experiments. Upregulated expression of the pH-sensitive kinase Pyk2 in OGR1 KO mouse proximal tubule cells may compensate for the loss of OGR1. Thus, we present the first evidence that OGR1 modulates the activity of two major plasma membrane proton transport systems. OGR1 may be involved in the regulation of plasma membrane transport proteins and intra- and/or extracellular pH.     

3-Acetyl-11-keto-β-boswellic acid attenuated oxidative glutamate toxicity in neuron-like cell lines by apoptosis inhibition

3-Acetyl-11-keto-β-boswellic acid (AKBA), a pentacyclic triterpenic acid present in gum resin of Boswellia serrata, has been found to possess antioxidant and neuroprotective properties. In this study, we aimed to examine protective properties of AKBA against glutamate-induced neuronal injury. To investigate the effects of AKBA (2.5-10 µM) on glutamate injury in neuron-like cells PC12 and N2a, two treatment regimens (incubation for 2 or 0 hours before glutamate exposure) were used. Then, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method was used to determine viability of the cells. Cellular redox status was evaluated using fluorimetry and comet assays. Annexin V/propidium iodide double staining and Western blot analysis of relative apoptotic proteins were conducted. Based on the results, 24 hours incubation with glutamate (8 mM) increased the cell mortality of PC12 and N2a (P < .001). However, AKBA (2.5-10 µM) enhanced the cell viability in both treatment regimens (P < .001). Also co- and pretreatment with AKBA significantly attenuated lipid peroxidation, reactive oxygen species production, and DNA injury (P < .05 and P < .001). AKBA also restored the activity of cellular superoxide dismutase under glutamate toxicity; this effect was seen to be more significant during the pretreatment regimen (P < .001). Moreover, Western blot analysis indicated that AKBA inhibited glutamate-induced programmed cell death through depressing the elevation of the expression ratio of Bax/Bcl-2 and cleaved-caspase-3 proteins, concentration-dependently. Overall, the present findings suggest the neuroprotective activities of AKBA against glutamate-induced cell injury probably by inhibiting oxidative damage and reducing apoptotic cell death.     

Predictive value of weight-for-age to identify overweight children

Objective: The objective was to assess the predictive value of weight‐for‐age to identify overweight children and adolescents in the unusual research or public health situations where height is not available to calculate BMI. Research Methods and Procedures: Data from the National Health and Nutrition Examination Survey 1999 to 2004 were used to calculate the sensitivity, specificity, and positive and negative predictive values of selected weight‐for‐age cut‐off points to identify overweight children and adolescents (as defined by BMI ≥95th percentile). Positive and negative predictive values are dependent on prevalence and are reported here for this study population only. Results: The 50th and 75th weight‐for‐age percentiles had good sensitivity (100% and 99.6%, respectively), but poor positive predictive value (23.7% and 37.0%, respectively), while the 95th and 97th percentiles had reasonable positive predictive value (80.3% and 91.5%, respectively), but limited sensitivity (82.0% and 66.7%, respectively) to identify overweight subjects. The properties of weight‐for‐age percentiles to identify overweight subjects differed between sex, age, and race/ethnicity but remain within a relatively narrow range. Discussion: No single weight‐for‐age cut‐off point was found to identify overweight children and adolescents with acceptable values for all properties and, therefore, cannot be used in the clinical setting. Furthermore, the positive predictive values reported here may be lower in populations with a lower prevalence of obesity. However, in unusual research or public health situations where height is not available, such as existing databases, weight‐for‐age percentiles may be useful to target limited resources to groups more likely to include overweight children and adolescents than the general population.     

Limber pine (Pinus flexilis James) genetic map constructed by exome‐seq provides insight into the evolution of disease resistance and a genomic resource for genomics‐based breeding

Limber pine ( Pinus flexilis ) is a keystone species of high‐elevation forest ecosystems of western North America, but some parts of the geographic range have high infection and mortality from the non‐native white pine blister rust caused by Cronartium ribicola . Genetic maps can provide essential knowledge for understanding genetic disease resistance as well as local adaptation to changing climates. Exome‐seq was performed to construct high‐density genetic maps in two seed families. Composite maps positioned 9612 unigenes across 12 linkage groups ( LG s). Syntenic analysis of genome structure revealed that the majority of orthologs were positional orthologous genes ( POG s) with localization on homologous LG s among conifer species. Gene ontology ( GO) enrichment analysis showed relatively fewer constraints for POG s with putative roles in adaptation to environments and relatively more conservation for POG s with roles in basic cell function and maintenance. The mapped genes included 639 nucleotide‐binding site leucine‐rich repeat genes ( NBS ‐ LRR s) , 290 receptor‐like protein kinase genes ( RLK s), and 1014 genes with potential roles in the defense response and induced systemic resistance to attack by pathogens. Orthologous loci for resistance to rust pathogens were identified and were co‐positioned with multiple members of the R gene family, revealing the evolutionary pressure acting upon them. This high‐density genetic map provides a genomic resource and practical tool for breeding and genetic conservation programs, with applications in genome‐wide association studies ( GWASs ), the characterization of functional genes underlying complex traits, and the sequencing and assembly of the full‐length genomes of limber pine and related Pinus species.     

Powder formulation of Burkholderia cepacia for control of rape seed damping-off caused by Rhizoctonia solani

Talc-based formulation of Burkholderia cepaci strain Bu1 was tested as seed and soil drenchs separately for its ability to control Rhizoctonia soloni the causal agent of rape seed damping-off in greenhouse and field trials. In general, the formulated bacteria was more effective to suppress the disease than the suspension of bacteria cells in carboxymethylcellulose solution (1% w/v), in both greenhouse and field trials. The formulation of strain Bul as soil and seed treatments had the greatest effect on reducing the rape seed damping-off in greenhouse and field trials (66.7, 53.3, 64.4 and 40% respectively). The formulation of strain Bu1 as soil and seed treatments were the most effective treatments to increase the root dry weights in the infected soil in greenhouse. The formulation of strain Bul as soil drench had the greatest effect on enhancement of the fresh weight of roots and stem fresh and dry weights. The formulation of strain Bu1 stored at 4 degrees C exhibited better shelf Life and efficacy in vitro than it's counterpart stored at 25 degrees C.     

Biological control of Sclerotinia sclerotiorum (Lib.) de Bary, the causal agent of white mold, by Pseudomonas species on canola petals

Sclerotinia sclerotiorum is an important pathogen on canola. Due to the public concern over pesticide use, alternative methods of disease control, such as biological control, should be considered. Several bacterial strains were isolated from canola and soja plants. Inhibition of S. sclerotiorum by bacterial strains in vitro was assayed on PDA medium in dual culture test. Eight Pseudomonas sp. strains (PB-3, PB-4, PB-5, PB-6, PB-7, PB-8, PB-10 and PB-11) caused inhibition zone against 5. sclerotiorum hyphal growth. The biocontrol potential of the bacteria was tested in a plant assay. Disease suppression was investigated using a petal inoculation technique. Canola petals were pretreated with bacteria, and then inoculated with 5. sclerotiorum ascospores 24 h later. Greenhouse experiment showed that application of Pseudomonas sp. strains (1 x 10(8) cfu ml(-1)) effectively suppressed S. sclerotiorum (1 x 10(5) ascospores ml(-1)) on petals and all of them achieved significant (P<0.01) disease suppression. Fourteen days after inoculation, strain PB-3 had 88/7% disease control and strain PB-4 had 69/9% disease control. Result from all studies indicates PB-3 to be effective biocontrol against S. sclerotiorum of canola. PB-3, PB-4, PB-7, PB-8, PB-10 and PB-11 were identified as Pseudomonas fluorescens biovar III. PB-5 and PB-6 was identified as Pseudomonas fluorescens biovar II. Strains PB-3, PB-4, PB-6, PB-10 and PB-11 produced protease and HCN. Strain PB-5 produce protease; no HCN.     

Biocontrol of Phytophthora cactorum the causal agent of root and crown rot on apple (Malus domestica) by formulated Pseudomonas fluorescens

In this study 284 isolates were isolated of apple trees' rhizosphere from Iran and 128 isolates were obtained from the collection of Research Department of Biological Control of University of Tehran. Four strains (P60, P61, P96, and P97) of Pseudomonas fluorescens were selected for greenhouse trials. The results of greenhouse trials showed dipping the crown and root of apple seedlings (MM106) combined with soil drench was more effective than dipping the crown and root on reducing the disease. After 6 weeks, strain P60 in dipping method combined with soil drench with 70% control, exhibited greatest effect on reducing the crown and root rot and was more effective than the fungicide metalaxyl-mancozeb. After 12 weeks, strains P60 and P96 in dipping method combined with soil drench with 55.6% and 44.5% control respectively, exhibited greatest effects on reducing the diseases Study of media on growth rate populations of effective strains exhibited that the beet molasses yeast extract (1:1) had more effect than nutrient broth(NB) medium. The initial high populations of powder formulations of strains P60 and P96 decreased during the storage at 4 and 25 degrees C over a 150-day period. In addition, formulations of strains stored at 4 degrees C had longer shelf life than those stored at 25 degrees C. In glasshouse trials, after 6 weeks, formulation of strain P60 and unformulated P60, obtained from NB medium and formulated P60, obtained from molasses yeast extract medium, and metalaxyl-mancozeb had highest effect on reducing the disease on apple rootstocks. After 12 weeks, formulation of strain P60 and unformulated bacteria obtained from both media, and metalaxyl-mancozeb with 57.1% control showed greatest effect on reducing the disease.