Differential localization and anabolic responsiveness of mTOR complexes in human skeletal muscle in response to feeding and exercise

Mechanistic target of rapamycin (mTOR) resides as two complexes within skeletal muscle. mTOR complex 1 (mTORC1-Raptor positive) regulates skeletal muscle growth, whereas mTORC2 (Rictor positive) regulates insulin sensitivity. To examine the regulation of these complexes in human skeletal muscle, we utilised immunohistochemical analysis to study the localisation of mTOR complexes prior to and following protein-carbohydrate feeding (FED) and resistance exercise plus protein-carbohydrate feeding (EXFED) in unilateral exercise model. In basal samples, mTOR and the lysosomal marker LAMP2 were highly co-localized and remained so throughout. In the FED and EXFED states, mTOR/LAMP2 complexes were redistributed to the cell periphery (WGA positive staining) (time effect; p=.025), with 39\% (FED) and 26\% (EXFED) increases in mTOR/WGA association observed 1h post-feeding/exercise. mTOR/WGA colocalisation continued to increase in EXFED at 3h (48\% above baseline) whereas colocalisation decreased in FED (21\% above baseline). A significant effect of condition (p=.05) was noted suggesting mTOR/WGA co-localization was greater during EXFED. This pattern was replicated in Raptor/WGA association, where a significant difference between EXFED and FED was noted at 3h post-exercise/feeding (p=.014). Rictor/WGA colocalization remained unaltered throughout the trial. Alterations in mTORC1 cellular location coincided with elevated S6K1 kinase activity, which rose to a greater extent in EXFED compared to FED at 1h post-exercise/feeding (p<.001), and only remained elevated in EXFED at the 3h time point (p=.037). Collectively these data suggest that mTORC1 redistribution within the cell is a fundamental response to resistance exercise and feeding, whereas mTORC2 is predominantly situated at the sarcolemma and does not alter localisation

Epigenetic gene regulation by Janus kinase 1 in diffuse large B-cell lymphoma

Janus kinases (JAKs) classically signal by activating STAT transcription factors but can also regulate gene expression by epigenetically phosphorylating histone H3 on tyrosine 41 (H3Y41-P). In diffuse large B-cell lymphomas (DLBCLs), JAK signaling is a feature of the activated B-cell (ABC) subtype and is triggered by autocrine production of IL-6 and IL-10. Whether this signaling involves STAT activation, epigenetic modification of chromatin, or both mechanisms is unknown. Here we use genetic and pharmacological inhibition to show that JAK1 signaling sustains the survival of ABC DLBCL cells. Whereas STAT3 contributed to the survival of ABC DLBCL cell lines, forced STAT3 activity could not protect these cells from death following JAK1 inhibition, suggesting epigenetic JAK1 action. JAK1 regulated the expression of nearly 3,000 genes in ABC DLBCL cells, and the chromatin surrounding many of these genes was modified by H3Y41-P marks that were diminished by JAK1 inhibition. These JAK1 epigenetic target genes encode important regulators of ABC DLBCL proliferation and survival, including IRF4, MYD88, and MYC. A small molecule JAK1 inhibitor cooperated with the BTK inhibitor ibrutinib in reducing IRF4 levels and acted synergistically to kill ABC DLBCL cells, suggesting that this combination should be evaluated in clinical trials.This research was supported by the Intramural Research Program of the NIH National Cancer Institute, the University of Wisconsin–Madison (UW–Madison) start-up funds, KL2 Scholar Awards UL1TR0000427 and KL2TR000428, the National Cancer Institute Grant 1R01 CA187299 (to L.R.), and the UW–Madison T32 Hematology Training Award T32 HL07899 (to A.C.D.)

Effect of arsenic-phosphorus interaction on arsenic-induced oxidative stress in chickpea plants

Arsenic-induced oxidative stress in chickpea was investigated under glasshouse conditions in response to application of arsenic and phosphorus. Three levels of arsenic (0, 30 and 60 mg kg−1) and four levels of P (50, 100, 200, and 400 mg kg−1) were applied to soil-grown plants. Increasing levels of both arsenic and P significantly increased arsenic concentrations in the plants. Shoot growth was reduced with increased arsenic supply regardless of applied P levels. Applied arsenic induced oxidative stress in the plants, and the concentrations of H2O2 and lipid peroxidation were increased. Activity of superoxide dismutase (SOD) and concentrations of non-enzymatic antioxidants decreased in these plants, but activities of catalase (CAT) and ascorbate peroxidase (APX) were significantly increased under arsenic phytotoxicity. Increased supply of P decreased activities of CAT and APX, and decreased concentrations of non-enzymatic antioxidants, but the high-P plants had lowered lipid peroxidation. It can be concluded that P increased uptake of arsenic from the soil, probably by making it more available, but although plant growth was inhibited by arsenic the P may have partially protected the membranes from arsenic-induced oxidative stress

Resistance exercise initiates mechanistic target of rapamycin (mTOR) translocation and protein complex co-localisation in human skeletal muscle

This is the final version. Available from the publisher via the DOI in this record.The mechanistic target of rapamycin (mTOR) is a central mediator of protein synthesis in skeletal muscle. We utilized immunofluorescence approaches to study mTOR cellular distribution and protein-protein co-localisation in human skeletal muscle in the basal state as well as immediately, 1 and 3 h after an acute bout of resistance exercise in a fed (FED; 20 g Protein/40 g carbohydrate/1 g fat) or energy-free control (CON) state. mTOR and the lysosomal protein LAMP2 were highly co-localised in basal samples. Resistance exercise resulted in rapid translocation of mTOR/LAMP2 towards the cell membrane. Concurrently, resistance exercise led to the dissociation of TSC2 from Rheb and increased in the co-localisation of mTOR and Rheb post exercise in both FED and CON. In addition, mTOR co-localised with Eukaryotic translation initiation factor 3 subunit F (eIF3F) at the cell membrane post-exercise in both groups, with the response significantly greater at 1 h of recovery in the FED compared to CON. Collectively our data demonstrate that cellular trafficking of mTOR occurs in human muscle in response to an anabolic stimulus, events that appear to be primarily influenced by muscle contraction. The translocation and association of mTOR with positive regulators (i.e. Rheb and eIF3F) is consistent with an enhanced mRNA translational capacity after resistance exercise.Biotechnology and Biological Science Research Council (BBSRC)Natural Sciences and Engineering Research Council (NSERC)China Scholarship CouncilNational Institute of Arthritis and Musculoskeletal and Skin DiseasesDepartment of Defens

Wheat rust epidemics damage Ethiopian wheat production: A decade of field disease surveillance reveals national-scale trends in past outbreaks.

Wheat rusts are the key biological constraint to wheat production in Ethiopia-one of Africa's largest wheat producing countries. The fungal diseases cause economic losses and threaten livelihoods of smallholder farmers. While it is known that wheat rust epidemics have occurred in Ethiopia, to date no systematic long-term analysis of past outbreaks has been available. We present results from one of the most comprehensive surveillance campaigns of wheat rusts in Africa. More than 13,000 fields have been surveyed during the last 13 years. Using a combination of spatial data-analysis and visualization, statistical tools, and empirical modelling, we identify trends in the distribution of wheat stem rust (Sr), stripe rust (Yr) and leaf rust (Lr). Results show very high infection levels (mean incidence for Yr: 44%; Sr: 34%; Lr: 18%). These recurrent rust outbreaks lead to substantial economic losses, which we estimate to be of the order of 10s of millions of US-D annually. On the widely adopted wheat variety, Digalu, there is a marked increase in disease prevalence following the incursion of new rust races into Ethiopia, which indicates a pronounced boom-and-bust cycle of major gene resistance. Using spatial analyses, we identify hotspots of disease risk for all three rusts, show a linear correlation between altitude and disease prevalence, and find a pronounced north-south trend in stem rust prevalence. Temporal analyses show a sigmoidal increase in disease levels during the wheat season and strong inter-annual variations. While a simple logistic curve performs satisfactorily in predicting stem rust in some years, it cannot account for the complex outbreak patterns in other years and fails to predict the occurrence of stripe and leaf rust. The empirical insights into wheat rust epidemiology in Ethiopia presented here provide a basis for improving future surveillance and to inform the development of mechanistic models to predict disease spread

Metabolites identified during varied doses of aspergillus species in Zea mays grains, and their correlation with aflatoxin levels

Open Access Journal; Published online: 7 May 2018Aflatoxin contamination is associated with the development of aflatoxigenic fungi such as Aspergillus flavus and A. parasiticus on food grains. This study was aimed at investigating metabolites produced during fungal development on maize and their correlation with aflatoxin levels. Maize cobs were harvested at R3 (milk), R4 (dough), and R5 (dent) stages of maturity. Individual kernels were inoculated in petri dishes with four doses of fungal spores. Fungal colonisation, metabolite profile, and aflatoxin levels were examined. Grain colonisation decreased with kernel maturity: milk-, dough-, and dent-stage kernels by approximately 100%, 60%, and 30% respectively. Aflatoxin levels increased with dose at dough and dent stages. Polar metabolites including alanine, proline, serine, valine, inositol, iso-leucine, sucrose, fructose, trehalose, turanose, mannitol, glycerol, arabitol, inositol, myo-inositol, and some intermediates of the tricarboxylic acid cycle (TCA—also known as citric acid or Krebs cycle) were important for dose classification. Important non-polar metabolites included arachidic, palmitic, stearic, 3,4-xylylic, and margaric acids. Aflatoxin levels correlated with levels of several polar metabolites. The strongest positive and negative correlations were with arabitol (R = 0.48) and turanose and (R = 0.53), respectively. Several metabolites were interconnected with the TCA; interconnections of the metabolites with the TCA cycle varied depending upon the grain maturity

Photoswitchable diacylglycerols enable optical control of protein kinase C.

Increased levels of the second messenger lipid diacylglycerol (DAG) induce downstream signaling events including the translocation of C1-domain-containing proteins toward the plasma membrane. Here, we introduce three light-sensitive DAGs, termed PhoDAGs, which feature a photoswitchable acyl chain. The PhoDAGs are inactive in the dark and promote the translocation of proteins that feature C1 domains toward the plasma membrane upon a flash of UV-A light. This effect is quickly reversed after the termination of photostimulation or by irradiation with blue light, permitting the generation of oscillation patterns. Both protein kinase C and Munc13 can thus be put under optical control. PhoDAGs control vesicle release in excitable cells, such as mouse pancreatic islets and hippocampal neurons, and modulate synaptic transmission in Caenorhabditis elegans. As such, the PhoDAGs afford an unprecedented degree of spatiotemporal control and are broadly applicable tools to study DAG signaling

Coupled climate response to Atlantic Multidecadal Variability in a multi-model multi-resolution ensemble

North Atlantic sea surface temperatures (SSTs) underwent pronounced multidecadal variability during the twentieth and early twenty-first century. We examine the impacts of this Atlantic Multidecadal Variability (AMV), also referred to as the Atlantic Multidecadal Oscillation (AMO), on climate in an ensemble of five coupled climate models at both low and high spatial resolution. We use a SST nudging scheme specified by the Coupled Model Intercomparision Project’s Decadal Climate Prediction Project Component C (CMIP6 DCPP-C) to impose a persistent positive/negative phase of the AMV in the North Atlantic in coupled model simulations; SSTs are free to evolve outside this region. The large-scale seasonal mean response to the positive AMV involves widespread warming over Eurasia and the Americas, with a pattern of cooling over the Pacific Ocean similar to the Pacific Decadal Oscillation (PDO), together with a northward displacement of the inter-tropical convergence zone (ITCZ). The accompanying changes in global atmospheric circulation lead to widespread changes in precipitation. We use Analysis of Variance (ANOVA) to demonstrate that this large-scale climate response is accompanied by significant differences between models in how they respond to the common AMV forcing, particularly in the tropics. These differences may arise from variations in North Atlantic air-sea heat fluxes between models despite a common North Atlantic SST forcing pattern. We cannot detect a widespread effect of increased model horizontal resolution in this climate response, with the exception of the ITCZ, which shifts further northwards in the positive phase of the AMV in the higher resolution configuratio