Increasing leucine concentration stimulates mechanistic target of rapamycin signaling and cell growth in C2C12 skeletal muscle cells

Leucine is a key amino acid for initiating translation in muscle cells, but the dose-dependent effects of leucine on intracellular signaling are poorly characterized. This study examined the effect that increasing doses of leucine would have on changes in mechanistic target of rapamycin (mTOR)–mediated signaling, rates of protein synthesis, and cell size in C2C12 cells. We hypothesized that a leucine “threshold” exists, which represents the minimum stimulus required to initiate mTOR signaling in muscle cells. Acute exposure to 1.5, 3.2, 5.0, and 16.1 mM leucine increased phosphorylation of mTORSer2448 (~1.4-fold; P < .04), 4E-BP1 Thr37/46 (~1.9-fold; P < .001), and rpS6Ser235/6 (~2.3-fold; P < .001). However, only p70S6kThr389 exhibited a dose-dependent response to leucine with all treatments higher than control (~4-fold; P < .001) and at least 5 mM higher than the 1.5-mM concentration (1.2-fold; P < .02). Rates of protein synthesis were not altered by any treatment. Seven days of exposure to 0.5, 1.5, 5.0, and 16.5 mM leucine resulted in an increase in cell size in at least 5 mM treatments (~1.6-fold, P < .001 vs control). Our findings indicate that even at low leucine concentrations, phosphorylation of proteins regulating translation initiation signaling is enhanced. The phosphorylation of p70S6kThr389 follows a leucine dose-response relationship, although this was not reflected by the acute protein synthetic response. Nevertheless, under the conditions of the present study, it appears that leucine concentrations of at least 5 mM are necessary to enhance cell growth.

Ibuprofen supplementation and its effects on NF-KB activation in skeletal muscle following resistance exercise

Resistance exercise triggers a subclinical inflammatory response that plays a pivotal role in skeletal muscle regeneration. Nuclear factor‐κB (NF‐κB) is a stress signalling transcription factor that regulates acute and chronic states of inflammation. The classical NF‐κB pathway regulates the early activation of post‐exercise inflammation; however there remains scope for this complex transcription factor to play a more detailed role in post‐exercise muscle recovery. Sixteen volunteers completed a bout of lower body resistance exercise with the ingestion of three 400 mg doses of ibuprofen or a placebo control. Muscle biopsy samples were obtained prior to exercise and at 0, 3 and 24 h post‐exercise and analysed for key markers of NF‐κB activity. Phosphorylated p65 protein expression and p65 inflammatory target genes were elevated immediately post‐exercise independent of the two treatments. These changes did not translate to an increase in p65 DNA binding activity. NF‐κB p50 protein expression and NF‐κB p50 binding activity were lower than pre‐exercise at 0 and 3 h post‐exercise, but were elevated at 24 h post‐exercise. These findings provide novel evidence that two distinct NF‐κB pathways are active in skeletal muscle after resistance exercise. The initial wave of activity involving p65 resembles the classical pathway and is associated with the onset of an acute inflammatory response. The second wave of NF‐κB activity comprises the p50 subunit, which has been previously shown to resolve an acute inflammatory program. The current study showed no effect of the ibuprofen treatment on markers of the NF‐κB pathway, however examination of the within group effects of the exercise protocol suggests that this pathway warrants further research.

Altering the redox state of skeletal muscle by glutathione depletion increases the exercise-activation of PGC-1α

We investigated the relationship between mitochondrial biogenesis, cell signalling and antioxidant enzymes by depleting skeletal muscle glutathione with diethyl maleate (DEM) which resulted in a demonstrable increase in oxidative stress during exercise. Animals were divided into six groups: (1) sedentary control rats; (2) sedentary rats treated with DEM; (3) exercise control rats euthanized immediately after exercise; (4) exercise rats + DEM; (5) exercise control rats euthanized 4 h after exercise, and; (6) exercise rats + DEM euthanized 4 h after exercise. Exercising animals ran on the treadmill at a 10% gradient at 20 m/min for the first 30 min. The speed was then increased every 10 min by 1.6 m/min until exhaustion. There was a reduction in total glutathione in the skeletal muscle of DEM treated animals compared to the control animals (P<0.05). Within the control group, total glutathione was higher in the sedentary group compared to after exercise (P<0.05). DEM treatment also significantly increased oxidative stress, as measured by increased plasma F2-isoprostanes (P<0.05). Exercising animals given DEM showed a significantly greater increase in peroxisome proliferator activated receptor γ coactivator-1α(PGC-1α) mRNA compared to the control animals that were exercised (P<0.05). This study provides novel evidence that by reducing the endogenous antioxidant glutathione in skeletal muscle and inducing oxidative stress through exercise, PGC-1α gene expression was augmented. These findings further highlight the important role of exercise induced oxidative stress in the regulation of mitochondrial biogenesis.

Mitochondrial genome acquisition restores respiratory function and tumorigenic potential of cancer cells without mitochondrial DNA

We report that tumor cells devoid of their mitochondrial genome (mtDNA) show delayed tumor growth and that tumor formation is associated with acquisition of mtDNA from host cells. This leads to partial recovery of mitochondrial function in cells derived from primary tumors grown from cells without mtDNA and a shorter lag in tumor growth. Cell lines from circulating tumor cells showed further recovery of mitochondrial respiration and an intermediate lag to tumor growth, while cells from lung metastases exhibited full restoration of respiratory function and no lag in tumor growth. Stepwise assembly of mitochondrial respiratory supercomplexes was correlated with acquisition of respiratory function. Our findings indicate horizontal transfer of mtDNA from host cells in the tumor microenvironment to tumor cells with compromised respiratory function to re-establish respiration and tumor-initiating efficacy. These results suggest a novel pathophysiological process for overcoming mtDNA damage and support the notion of high plasticity of malignant cells.

The ethical intricacies of injury rehabilitation within a dance training context

Exploring the ethical issues present in professional practice within the field of sport, exercise and performance psychology, this case study outlines challenges that may be encountered, ways to address issues should they arise, and the overall ethical considerations of supporting injury rehabilitation within a dance training context.

Cytokine expression and secretion by skeletal muscle cells : regulatory mechanisms and exercise effects

Cytokines are important mediators of various aspects of health and disease, including appetite, glucose and lipid metabolism, insulin sensitivity, skeletal muscle hypertrophy and atrophy. Over the past decade or so, considerable attention has focused on the potential for regular exercise to counteract a range of disease states by modulating cytokine production. Exercise stimulates moderate to large increases in the circulating concentrations of interleukin (IL)-6, IL-8, IL-10, IL-1 receptor antagonist, granulocyte-colony stimulating factor, and smaller increases in tumor necrosis factor-α, monocyte chemotactic protein-1, IL-1β, brain-derived neurotrophic factor, IL-12p35/p40 and IL-15. Although many of these cytokines are also expressed in skeletal muscle, not all are released from skeletal muscle into the circulation during exercise. Conversely, some cytokines that are present in the circulation are not expressed in skeletal muscle after exercise. The reasons for these discrepant cytokine responses to exercise are unclear. In this review, we address these uncertainties by summarizing the capacity of skeletal muscle cells to produce cytokines, analyzing other potential cellular sources of circulating cytokines during exercise, and discussing the soluble factors and intracellular signaling pathways that regulate cytokine synthesis (e.g., RNA-binding proteins, microRNAs, suppressor of cytokine signaling proteins, soluble receptors).

Proactive training system for safe and efficient precast installation

The construction industry is a crucial component of the Hong Kong economy, and the safety and efficiency of workers are two of its main concerns. The current approach to training workers relies primarily on instilling practice and experience in conventional teacher-apprentice settings on and off site. Both have their limitations however, on-site training is very inefficient and interferes with progress on site, while off-site training provides little opportunity to develop the practical skills and awareness needed through hands-on experience. A more effective way is to train workers in safety awareness and efficient working by current novel information technologies. This paper describes a new and innovative prototype system – the Proactive Construction Management System (PCMS) – to train precast installation workers to be highly productive while being fully aware of the hazards involved. PCMS uses Chirp-Spread-Spectrum-based (CSS) real-time location technology and Unity3D-based data visualisation technology to track construction resources (people, equipment, materials, etc.) and provide real-time feedback and post-event visualisation analysis in a training environment. A trial of a precast facade installation on a real site demonstrates the benefits gained by PCMS in comparison with equivalent training using conventional methods. It is concluded that, although the study is based on specific industrial conditions found in Hong Kong construction projects, PCMS may well attract wider interest and use in future.

The protective mechanisms of first aid training within a school-based program for early adolescents

This thesis was concerned with the protective mechanisms of first aid training in the context of peer support. Using a randomised control trial design the current program of research explores first aid training in the school setting and identifies the key components of effective school-based first aid training programs. In particular, examining whether first aid training and associated knowledge could be protective for early adolescents. This broader framing considered whether first aid impacted on increasing behaviour and attitudes towards helping an injured friend, and reducing personal risk taking and related injury.

Regressive measures won’t help youth into work or training

The 2014 federal budget implemented a so-called crackdown on what Minister for Social Services Kevin Andrews calls young people who are content to “sit on the couch at home and pick up a welfare cheque”. The crackdown will change access to income support for people under 30 years of age. From January 1 2015, all young people seeking Newstart Allowance and Youth Allowance for the first time will be required to “demonstrate appropriate job search and participation in employment services support for six months before receiving payments”. Upon qualifying, recipients must then spend 25 hours per week in Work for the Dole in order to receive income support for a six-month period. What happens beyond this six months is unclear. What is clear is that these policy changes, together with the Minister’s accompanying statements, are informed by a deficit view of disadvantaged youth. It is a view that demonstrates how little politicians know or understand about these young peoples’ past circumstances.

How do experts think about statistics? Hints for improving undergraduate and postgraduate training

Experts are increasingly being called upon to quantify their knowledge, particularly in situations where data is not yet available or of limited relevance. In many cases this involves asking experts to estimate probabilities. For example experts, in ecology or related fields, might be called upon to estimate probabilities of incidence or abundance of species, and how they relate to environmental factors. Although many ecologists undergo some training in statistics at undergraduate and postgraduate levels, this does not necessarily focus on interpretations of probabilities. More accurate elicitation can be obtained by training experts prior to elicitation, and if necessary tailoring elicitation to address the expert’s strengths and weaknesses. Here we address the first step of diagnosing conceptual understanding of probabilities. We refer to the psychological literature which identifies several common biases or fallacies that arise during elicitation. These form the basis for developing a diagnostic questionnaire, as a tool for supporting accurate elicitation, particularly when several experts or elicitors are involved. We report on a qualitative assessment of results from a pilot of this questionnaire. These results raise several implications for training experts, not only prior to elicitation, but more strategically by targeting them whilst still undergraduate or postgraduate students.

The QUT Training Model for Peer Leader Capacity Building

QUT has enacted a university-wide Peer Program’s Strategy which aims to improve student success and graduate outcomes. A component of this strategy is a training model providing relevant, quality-assured and timely training for all students who take on leadership roles. The training model is designed to meet the needs of the growing scale and variety of peer programs, and to recognise the multiple roles and programs in which students may be involved during their peer leader journey. The model builds peer leader capacity by offering centralised, beginning and ongoing training modules, delivered by in-house providers, covering topics which prepare students to perform their role safely, inclusively, accountably and skilfully. The model also provides efficiencies by differentiating between ‘core competency' and ‘program-specific’ modules, thus avoiding training duplication across multiple programs, and enabling training to be individually and flexibly formatted to suit the specific and unique needs of each program.