LAT1 Protein Content Increases Following 12 Weeks of Resistance Exercise Training in Human Skeletal Muscle

Introduction: Amino acid transporters are essential for cellular amino acid transport and promoting protein synthesis. While previous literature has demonstrated the association of amino acid transporters and protein synthesis following acute resistance exercise and amino acid supplementation, the chronic effect of resistance exercise and supplementation on amino acid transporters is unknown. The purpose herein was to determine if amino acid transporters and amino acid metabolic enzymes were related to skeletal muscle hypertrophy following resistance exercise training with different nutritional supplementation strategies. Methods: 43 college-aged males were separated into a maltodextrin placebo (PLA, n = 12), leucine (LEU, n = 14), or whey protein concentrate (WPC, n = 17) group and underwent 12 weeks of total-body resistance exercise training. Each group\u27s supplement was standardized for total energy and fat, and LEU and WPC supplements were standardized for total leucine (6 g/d). Skeletal muscle biopsies were obtained prior to training and ~72 h following each subject\u27s last training session. Results: All groups increased type I and II fiber cross-sectional area (fCSA) following training (p \u3c 0.050). LAT1 protein increased following training (p \u3c 0.001) and increased more in PLA than LEU and WPC (p \u3c 0.050). BCKDHα protein increased and ATF4 protein decreased following training (p \u3c 0.001). Immunohistochemistry indicated total LAT1/fiber, but not membrane LAT1/fiber, increased with training (p = 0.003). Utilizing all groups, the change in ATF4 protein, but no other marker, trended to correlate with the change in fCSA (r = 0.314; p = 0.055); however, when regression analysis was used to delineate groups, the change in ATF4 protein best predicted the change in fCSA only in LEU (r2 = 0.322; p = 0.043). In C2C12 myoblasts, LAT1 protein overexpression caused a paradoxical decrease in protein synthesis levels (p = 0.002) and decrease in BCKDHα protein (p = 0.001). Conclusions: Amino acid transporters and metabolic enzymes are affected by resistance exercise training, but do not appear to dictate muscle fiber hypertrophy. In fact, overexpression of LAT1 in vitro decreased protein synthesis

Protein Supplementation Throughout 10 Weeks of Progressive Run Training Is Not Beneficial for Time Trial Improvement

Introduction: Protein supplementation is proposed to promote recovery and adaptation following endurance exercise. While prior literature demonstrates improved performance when supplementing protein during or following endurance exercise, chronic supplementation research is limited.Methods: Runners (VO2peak = 53.6 ± 8.9 ml/kg/min) were counter-balanced into a placebo group (PLA; n = 8) or protein group (PRO; n = 9) based on sex and VO2peak, and underwent 10 weeks of progressive endurance training. Prior to training, body composition, blood cell differentials, non-invasive mitochondrial capacity using near-infrared spectroscopy, and a 5 km treadmill time trial (TT) were evaluated. Progressive training then commenced (5–10% increase in weekly volume with a recovery week following 3 weeks of training) whereby PRO supplemented with 25 g of whey protein following workouts and prior to sleep (additional 50 g daily). PLA supplemented similarly with a < 1 g sugar pill per day. Following training, participants were reanalyzed for the aforementioned tests.Results: VO2peak and initial 5 km TT were not significantly different between groups. PRO consumed significantly more dietary protein throughout the training period (PRO = 132 g/d or 2.1 g/kg/day; PLA = 84 g/d or 1.2 g/kg/day). Running volume increased significantly over time, but was not significantly different between groups throughout training. Blood measures were unaltered with training or supplementation. Mitochondrial capacity trended toward improving over time (time p = 0.063) with no difference between groups. PLA increased lean mass 0.7 kg (p < 0.05) while PRO experienced infinitesimal change (−0.1 kg, interaction p = 0.049). PLA improved 5 km TT performance 6.4% (1 min 31 s), while PRO improved only 2.7% (40 s) (interaction p = 0.080).Conclusion: This is the first evidence to suggest long-term protein supplementation during progressive run training is not beneficial for runners

Character and environmental lability of cyanobacteria-derived dissolved organic matter

Autotrophic dissolved organic matter (DOM) is central to the carbon biogeochemistry of aquatic systems, and the full complexity of autotrophic DOM has not been extensively studied, particularly by high-resolution mass spectrometry (HRMS). Terrestrial DOM tends to dominate HRMS studies in freshwaters due to the propensity of such compounds to ionize by negative mode electrospray, and possibly also because ionizable DOM produced by autotrophy is decreased to low steady-state concentrations by heterotrophic bacteria. In this study, we investigated the character of DOM produced by the widespread cyanobacteriaMicrocystis aeruginosausing high-pressure liquid chromatography-electrospray ionization-high-resolution mass spectrometry.M. aeruginosaproduced thousands of detectable compounds in axenic culture. These compounds were chromatographically resolved and the majority were assigned to aliphatic formulas with a broad polarity range. We found that the DOM produced byM. aeruginosawas highly susceptible to removal by heterotrophic freshwater bacteria, supporting the hypothesis that this autotroph-derived organic material is highly labile and accordingly only seen at low concentrations in natural settings

Dysfunction of Nrf-2 in CF Epithelia Leads to Excess Intracellular H2O2 and Inflammatory Cytokine Production

Cystic fibrosis is characterized by recurring pulmonary exacerbations that lead to the deterioration of lung function and eventual lung failure. Excessive inflammatory responses by airway epithelia have been linked to the overproduction of the inflammatory cytokine IL-6 and IL-8. The mechanism by which this occurs is not fully understood, but normal IL-1β mediated activation of the production of these cytokines occurs via H2O2 dependent signaling. Therefore, we speculated that CFTR dysfunction causes alterations in the regulation of steady state H2O2. We found significantly elevated levels of H2O2 in three cultured epithelial cell models of CF, one primary and two immortalized. Increases in H2O2 heavily contributed to the excessive IL-6 and IL-8 production in CF epithelia. Proteomic analysis of three in vitro and two in vivo models revealed a decrease in antioxidant proteins that regulate H2O2 processing, by ≥2 fold in CF vs. matched normal controls. When cells are stimulated, differential expression in CF versus normal is enhanced; corresponding to an increase in H2O2 mediated production of IL-6 and IL-8. The cause of this redox imbalance is a decrease by ∼70% in CF cells versus normal in the expression and activity of the transcription factor Nrf-2. Inhibition of CFTR function in normal cells produced this phenotype, while N-acetyl cysteine, selenium, an activator of Nrf-2, and the overexpression of Nrf-2 all normalized H2O2 processing and decreased IL-6 and IL-8 to normal levels, in CF cells. We conclude that a paradoxical decrease in Nrf-2 driven antioxidant responses in CF epithelia results in an increase in steady state H2O2, which in turn contributes to the overproduction of the pro-inflammatory cytokines IL-6 and IL-8. Treatment with antioxidants can ameliorate exaggerated cytokine production without affecting normal responses

Research Update on Extreme-Mass-Ratio Inspirals

The inspirals of stellar-mass mass compact objects into massive black holes in the centres of galaxies are one of the most important sources of gravitational radiation for space-based detectors like LISA or eLISA. These extreme-mass-ratio inspirals (EMRIs) will enable an ambitious research program with implications for astrophysics, cosmology, and fundamental physics. This article is a summary of the talks delivered at the plenary session on EMRIs at the 10th International LISA Symposium. It contains research updates on the following topics: astrophysics of EMRIs; EMRI science potential; and EMRI modeling.Comment: 17 pages, no figures. Proceedings of the LISA Symposium X, to be published at the Journal of Physic

A Versatile ΦC31 Based Reporter System for Measuring AP-1 and Nrf2 Signaling in Drosophila and in Tissue Culture

This paper describes the construction and characterization of a system of transcriptional reporter genes for monitoring the activity of signaling pathways and gene regulation mechanisms in intact Drosophila, dissected tissues or cultured cells. Transgenic integration of the reporters into the Drosophila germline was performed in a site-directed manner, using ΦC31 integrase. This strategy avoids variable position effects and assures low base level activity and high signal responsiveness. Defined integration sites furthermore enable the experimenter to compare the activity of different reporters in one organism. The reporter constructs have a modular design to facilitate the combination of promoter elements (synthetic transcription factor binding sites or natural regulatory sequences), reporter genes (eGFP, or DsRed.T4), and genomic integration sites. The system was used to analyze and compare the activity and signal response profiles of two stress inducible transcription factors, AP-1 and Nrf2. To complement the transgenic reporter fly lines, tissue culture assays were developed in which the same synthetic ARE and TRE elements control the expression of firefly luciferase

The Arctic in the twenty-first century: changing biogeochemical linkages across a paraglacial landscape of Greenland

The Kangerlussuaq area of southwest Greenland encompasses diverse ecological, geomorphic, and climate gradients that function over a range of spatial and temporal scales. Ecosystems range from the microbial communities on the ice sheet and moisture-stressed terrestrial vegetation (and their associated herbivores) to freshwater and oligosaline lakes. These ecosystems are linked by a dynamic glacio-fluvial-aeolian geomorphic system that transports water, geological material, organic carbon and nutrients from the glacier surface to adjacent terrestrial and aquatic systems. This paraglacial system is now subject to substantial change because of rapid regional warming since 2000. Here, we describe changes in the eco- and geomorphic systems at a range of timescales and explore rapid future change in the links that integrate these systems. We highlight the importance of cross-system subsidies at the landscape scale and, importantly, how these might change in the near future as the Arctic is expected to continue to warm