Vitamin D and the Athlete: Current Perspectives and New Challenges

The last decade has seen a dramatic increase in general interest in and research into vitamin D, with many athletes now taking vitamin D supplements as part of their everyday dietary regimen. The most recognized role of vitamin D is its regulation of calcium homeostasis; there is a strong relationship between vitamin D and bone health in non-athletic individuals. In contrast, data have consistently failed to demonstrate any relationship between serum 25[OH]D and bone health, which may in part be due to the osteogenic stimulus of exercise. Vitamin D may interact with extra-skeletal tissues such as muscle and the immune system to modulate recovery from damaging exercise and infection risk. Given that many athletes now engage in supplementation, often consuming extreme doses of vitamin D, it is important to assess whether excessive vitamin D can be detrimental to health. It has been argued that toxic effects only occur when serum 25[OH] D concentrations are greater than 180 nmol·l −1 , but data from our laboratory have suggested high-dose supplementation could be problematic. Finally, there is a paradoxical relationship between serum 25[OH]D concentration, ethnicity, and markers of bone health: Black athletes often present with low serum 25[OH] D without physiological consequences. One explanation for this could be genetic differences in vitamin D binding protein due to ethnicity, resulting in greater concentrations of bioavailable (or free) vitamin D in some ethnic groups. In the absence of any pathology, screening may be unnecessary and could result in incorrect supplementation. Data must now be re-examined, taking into consideration bioavailable or “free” vitamin D in ethnically diverse groups to enable new thresholds and target concentrations to be established; perhaps, for now, it is time to “set vitamin D free”. © 2018 The Author(s

MicroRNA-184 and its lncRNA sponge uca1 are induced in wounded keratinocytes in a store-operated calcium entry-dependent manner.

There remains a significant need for therapeutic interventions for wound healing as the clinical and socioeconomic challenges presented by the ageing population and associated rise in non-healing wounds persist, exerting enormous pressure on health services. MicroRNAs (miRNAs) are short ≈22 nucleotide non-coding RNA molecules that fine-tune gene expression by degradation of mRNA targets. Several miRNAs promote keratinocyte migration, including miR-21, miR-31 and miR-132 (reviewed in 3 ), hence miRNA-dependent migration is emerging as a framework for wound healing. This article is protected by copyright. All rights reserved

The Role of Vitamin D in Skeletal Muscle Function and Regeneration

Skeletal muscle is a direct target for the group of seco-steroids collectively termed Vitamin D. Skeletal muscle expresses both CYP27A1 and CYP27B1 encoding for the hydroxylases required to convert Vitamin D to 25[OH]D and subsequently the bioactive 1α-25-dihydroxyvitamin D3 (1α-25[OH]2D3) (Girgis et al., 2014b). Crucially, the Vitamin D receptor (VDR) is also present in skeletal muscle (Srikuea, Zhang, Park-Sarge, & Esser, 2012) and upon exposure, binds to its ligand 1α-25[OH]2D3 and initiates genomic and non-genomic rapid signalling responses. At present there is a global prevalence of low serum Vitamin D (25[OH]D) concentrations due to a lack of sun exposure (the primary route for Vitamin D synthesis) as a function of latitude and/or an indoor lifestyle coupled with few dietary sources of Vitamin D (Chen et al., 2007). Accumulating data are now suggestive that low 25[OH]D may be associated with perturbations in contractile activity and the regeneration of human skeletal muscle (Owens, Fraser, & Close, 2014), although a definitive causal relationship is yet to be established. Therefore, this thesis aimed to establish a more precise role for Vitamin D in human skeletal muscle function and regeneration. There were four overarching aims: 1.Explore the role of Vitamin D in human skeletal muscle contractile properties in humans in vivo. 2.Identify the role of Vitamin D in human skeletal muscle contractile properties ex vivo. 3.Investigate the role of Vitamin D in skeletal muscle regeneration following eccentric exercise induced muscle damage in vivo. 4.Elucidate cellular mechanisms of the muscle regeneration process that are responsive to Vitamin D in vitro. The main findings from this work imply that serum 25[OH]D concentrations across a broad range from 18->100 nmol.L-1 do not affect skeletal muscle contractile properties. Conversely elevating serum 25[OH]D from 75 nmol.L-1 resulted in significant improvements in the recovery of maximal voluntary contraction force following a bout of eccentric exercise. Implementing an in vitro model of muscle regeneration also identified potential cellular mechanisms for these observations: Muscle derived cells treated with a total amount of 10 nmol 1α-25[OH]2D3 following a mechanical scrape improved migration dynamics and fusion capability of skeletal muscle derived cells. Taken together, low Vitamin D concentrations are highly prevalent but can be easily corrected with supplementation of Vitamin D3. This may have the advantage of optimising the regenerative capacity of skeletal muscle amongst other health benefits previously characterised by others

Cross Pixel Optical Flow Similarity for Self-Supervised Learning

We propose a novel method for learning convolutional neural image representations without manual supervision. We use motion cues in the form of optical flow, to supervise representations of static images. The obvious approach of training a network to predict flow from a single image can be needlessly difficult due to intrinsic ambiguities in this prediction task. We instead propose a much simpler learning goal: embed pixels such that the similarity between their embeddings matches that between their optical flow vectors. At test time, the learned deep network can be used without access to video or flow information and transferred to tasks such as image classification, detection, and segmentation. Our method, which significantly simplifies previous attempts at using motion for self-supervision, achieves state-of-the-art results in self-supervision using motion cues, competitive results for self-supervision in general, and is overall state of the art in self-supervised pretraining for semantic image segmentation, as demonstrated on standard benchmarks

Skeletal muscle cells possess a 'memory' of acute early life TNF-α exposure: role of epigenetic adaptation.

Sufficient quantity and quality of skeletal muscle is required to maintain lifespan and healthspan into older age. The concept of skeletal muscle programming/memory has been suggested to contribute to accelerated muscle decline in the elderly in association with early life stress such as fetal malnutrition. Further, muscle cells in vitro appear to remember the in vivo environments from which they are derived (e.g. cancer, obesity, type II diabetes, physical inactivity and nutrient restriction). Tumour-necrosis factor alpha (TNF-α) is a pleiotropic cytokine that is chronically elevated in sarcopenia and cancer cachexia. Higher TNF-α levels are strongly correlated with muscle loss, reduced strength and therefore morbidity and earlier mortality. We have extensively shown that TNF-α impairs regenerative capacity in mouse and human muscle derived stem cells [Meadows et al. (J Cell Physiol 183(3):330-337, 2000); Foulstone et al. (J Cell Physiol 189(2):207-215, 2001); Foulstone et al. (Exp Cell Res 294(1):223-235, 2004); Stewart et al. (J Cell Physiol 198(2):237-247, 2004); Al-Shanti et al. (Growth factors (Chur, Switzerland) 26(2):61-73, 2008); Saini et al. (Growth factors (Chur, Switzerland) 26(5):239-253, 2008); Sharples et al. (J Cell Physiol 225(1):240-250, 2010)]. We have also recently established an epigenetically mediated mechanism (SIRT1-histone deacetylase) regulating survival of myoblasts in the presence of TNF-α [Saini et al. (Exp Physiol 97(3):400-418, 2012)]. We therefore wished to extend this work in relation to muscle memory of catabolic stimuli and the potential underlying epigenetic modulation of muscle loss. To enable this aim; C2C12 myoblasts were cultured in the absence or presence of early TNF-α (early proliferative lifespan) followed by 30 population doublings in the absence of TNF-α, prior to the induction of differentiation in low serum media (LSM) in the absence or presence of late TNF-α (late proliferative lifespan). The cells that received an early plus late lifespan dose of TNF-α exhibited reduced morphological (myotube number) and biochemical (creatine kinase activity) differentiation vs. control cells that underwent the same number of proliferative divisions but only a later life encounter with TNF-α. This suggested that muscle cells had a morphological memory of the acute early lifespan TNF-α encounter. Importantly, methylation of myoD CpG islands were increased in the early TNF-α cells, 30 population doublings later, suggesting that even after an acute encounter with TNF-α, the cells have the capability of retaining elevated methylation for at least 30 cellular divisions. Despite these fascinating findings, there were no further increases in myoD methylation or changes in its gene expression when these cells were exposed to a later TNF-α dose suggesting that this was not directly responsible for the decline in differentiation observed. In conclusion, data suggest that elevated myoD methylation is retained throughout muscle cells proliferative lifespan as result of early life TNF-α treatment and has implications for the epigenetic control of muscle loss

Efficacy of High Dose Vitamin D Supplements for Elite Athletes.

PURPOSE: Supplementation with dietary forms of vitamin D is commonplace in clinical medicine, elite athletic cohorts and the general population, yet the response of all major vitamin D metabolites to high doses of vitamin D is poorly characterized. We aimed to identify the responses of all major vitamin D metabolites to moderate and high dose supplemental vitamin D3. METHODS: A repeated measures design was implemented in which 46 elite professional European athletes were block randomized based on their basal 25[OH]D concentration into two treatment groups. Athletes received either 35,000 or 70,000 IU.week vitamin D3 for 12 weeks and 42 athletes completed the trial. Blood samples were collected over 18 weeks to monitor the response to supplementation and withdrawal from supplementation. RESULTS: Both doses led to significant increases in serum 25[OH]D and 1,25[OH]2D3. 70,000 IU.week also resulted in a significant increase of the metabolite 24,25[OH]2D at weeks 6 and 12 that persisted following supplementation withdrawal at week 18, despite a marked decrease in 1,25[OH]2D3. Intact PTH was decreased in both groups by week 6 and remained suppressed throughout the trial. CONCLUSIONS: High dose vitamin D3 supplementation (70,000 IU.week) may be detrimental for its intended purposes due to increased 24,25[OH]2D production. Rapid withdrawal from high dose supplementation may inhibit the bioactivity of 1,25[OH]2D3 as a consequence of sustained increases in 24,25[OH]2D that persist as 25[OH]D and 1,25[OH]2D concentrations decrease. These data imply that lower doses of vitamin D3 ingested frequently may be most appropriate and gradual withdrawal from supplementation as opposed to rapid withdrawal may be favorable

L-glutamine improves skeletal muscle cell differentiation and prevents myotube atrophy after cytokine (TNF-α) stress via reduced p38 MAPK signal transduction

Tumour Necrosis Factor- Alpha (TNF-α) is chronically elevated in conditions where skeletal muscle loss occurs. As L-glutamine can dampen the effects of inflamed environments, we investigated the role of L-glutamine in both differentiating C2C12 myoblasts and existing myotubes in the absence/presence of TNF-α (20 ng.ml−1) ± L-glutamine (20 mM).TNF-α reduced the proportion of cells in G1 phase, as well as biochemical (CK activity) and morphological differentiation (myotube number), with corresponding reductions in transcript expression of: Myogenin, Igf-I and Igfbp5. Furthermore, when administered to mature myotubes, TNF-α induced myotube loss and atrophy underpinned by reductions in Myogenin, Igf-I, Igfbp2 and glutamine synthetase and parallel increases in Fox03, Cfos, p53 and Bid gene expression. Investigation of signaling activity suggested that Akt and ERK1/2 were unchanged, JNK increased (non-significantly) whereas P38 MAPK substantially and significantly increased in both myoblasts and myotubes in the presence of TNF-α. Importantly, 20 mM L-glutamine reduced p38 MAPK activity in TNF-α conditions back to control levels, with a corresponding rescue of myoblast differentiation and a reversal of atrophy in myotubes. L-glutamine resulted in upregulation of genes associated with growth and survival including; Myogenin, Igf-Ir, Myhc2 & 7, Tnfsfr1b, Adra1d and restored atrophic gene expression of Fox03 back to baseline in TNF-α conditions. In conclusion, L-glutamine supplementation rescued suppressed muscle cell differentiation and prevented myotube atrophy in an inflamed environment via regulation of p38 MAPK. L-glutamine administration could represent an important therapeutic strategy for reducing muscle loss in catabolic diseases and inflamed ageing. This article is protected by copyright. All rights reserve

Lamin Mutations Cause Increased YAP Nuclear Entry in Muscle Stem Cells

Mutations in the LMNA gene, encoding the nuclear envelope A-type lamins, are responsible for muscular dystrophies, the most severe form being the LMNA-related congenital muscular dystrophy (L-CMD), with severe defects in myonucleus integrity. We previously reported that L-CMD mutations compromise the ability of muscle stem cells to modulate the yes-associated protein (YAP), a pivotal factor in mechanotransduction and myogenesis. Here, we investigated the intrinsic mechanisms by which lamins influence YAP subcellular distribution, by analyzing different conditions affecting the balance between nuclear import and export of YAP. In contrast to wild type (WT) cells, LMNADK32 mutations failed to exclude YAP from the nucleus and to inactivate its transcriptional activity at high cell density, despite activation of the Hippo pathway. Inhibiting nuclear pore import abolished YAP nuclear accumulation in confluent mutant cells, thus showing persistent nuclear import of YAP at cell confluence. YAP deregulation was also present in congenital myopathy related to nesprin-1KASH mutation, but not in cells expressing the LMNAH222P mutation, the adult form of lamin-related muscle dystrophy with reduced nuclear deformability. In conclusion, our data showed that L-CMD mutations increased YAP nuclear localization via an increased nuclear import and implicated YAP as a pathogenic contributor in muscle dystrophies caused by nuclear envelop defects

Admit your weakness: Verifying correctness on TSO architectures

“The final publication is available at http://link.springer.com/chapter/10.1007%2F978-3-319-15317-9_22 ”.Linearizability has become the standard correctness criterion for fine-grained non-atomic concurrent algorithms, however, most approaches assume a sequentially consistent memory model, which is not always realised in practice. In this paper we study the correctness of concurrent algorithms on a weak memory model: the TSO (Total Store Order) memory model, which is commonly implemented by multicore architectures. Here, linearizability is often too strict, and hence, we prove a weaker criterion, quiescent consistency instead. Like linearizability, quiescent consistency is compositional making it an ideal correctness criterion in a component-based context. We demonstrate how to model a typical concurrent algorithm, seqlock, and prove it quiescent consistent using a simulation-based approach. Previous approaches to proving correctness on TSO architectures have been based on linearizabilty which makes it necessary to modify the algorithm’s high-level requirements. Our approach is the first, to our knowledge, for proving correctness without the need for such a modification