Improving physiological relevance of cell culture: the possibilities, considerations, and future directions of the ex vivo coculture model.

In vitro models provide an important platform for the investigation of cellular growth and atrophy to inform, or extend mechanistic insights from, logistically challenging in vivo trials. Although these models allow for the identification of candidate mechanistic pathways, many models involve supraphysiological dosages, nonphysiological conditions, or experimental changes relating to individual proteins or receptors, all of which limit translation to human trials. To overcome these drawbacks, the use of ex vivo human plasma and serum has been used in cellular models to investigate changes in myotube hypertrophy, cellular protein synthesis, anabolic and catabolic markers in response to differing age, disease states, and nutrient status. However, there are currently no concurrent guidelines outlining the optimal methodology for this model. This review discusses the key methodological considerations surrounding the use of ex vivo plasma and serum with a focus in application to skeletal muscle cell lines (i.e., C2C12, L6, and LHCN-M2) and human primary skeletal muscle cells (HSMCs) as a means to investigate molecular signaling in models of atrophy and hypertrophy, alongside future directions.

High-impact jumping mitigates the short-term effects of low energy availability on bone resorption but not formation in regularly menstruating females: A randomized control trial.

Low energy availability (LEA) is prevalent in active individuals and negatively impacts bone turnover in young females. High-impact exercise can promote bone health in an energy efficient manner and may benefit bone during periods of LEA. Nineteen regularly menstruating females (aged 18-31 years) participated in two three-day conditions providing 15 (LEA) and 45 kcals kg fat-free mass-1 day-1 (BAL) of energy availability, each beginning 3 ± 1 days following the self-reported onset of menses. Participants either did (LEA+J, n = 10) or did not (LEA, n = 9) perform 20 high-impact jumps twice per day during LEA, with P1NP, ß-CTx (circulating biomarkers of bone formation and resorption, respectively) and other markers of LEA measured pre and post in a resting and fasted state. Data are presented as estimated marginal mean ± 95% CI. P1NP was significantly reduced in LEA (71.8 ± 6.1-60.4 ± 6.2 ng mL-1 , p < 0.001, d = 2.36) and LEA+J (93.9 ± 13.4-85.2 ± 12.3 ng mL-1 , p < 0.001, d = 1.66), and these effects were not significantly different (time by condition interaction: p = 0.269). ß-CTx was significantly increased in LEA (0.39 ± 0.09-0.46 ± 0.10 ng mL-1 , p = 0.002, d = 1.11) but not in LEA+J (0.65 ± 0.08-0.65 ± 0.08 ng mL-1 , p > 0.999, d = 0.19), and these effects were significantly different (time by condition interaction: p = 0.007). Morning basal bone formation rate is reduced following 3 days LEA, induced via dietary restriction, with or without high-impact jumping in regularly menstruating young females. However, high-impact jumping can prevent an increase in morning basal bone resorption rate and may benefit long-term bone health in individuals repeatedly exposed to such bouts.

The resting serum metabolome in response to short-term sprint interval training.

PURPOSE: To investigate the response of a targeted fraction of (168 metabolites) of the resting serum metabolome to 9 sessions of sprint interval training (SIT). METHODS: Thirty-four recreationally active males provided resting blood samples before (baseline) and 48-72 h after (post) a short-term (9 sessions) cycle ergometer-based SIT intervention. A targeted analysis of 168 metabolites was performed on serum using liquid chromatography mass spectrometry (LC-MS). 160 distinct metabolites were identified and combined with 4 calculated metabolite sums and 3 calculated metabolite ratios creating a panel of 167 individual factors. Data were analysed using principal component analysis and univariate testing of all factors classified into 5 metabolite subgroups. RESULTS: SIT improved anaerobic capacity measured by average power output during a Wingate test (p < 0.01; mean difference = 38 W, 95% confidence interval [26, 51]) and aerobic capacity measured by average power output in a 20 min cycling test (p < 0.01; 17 W [12, 23]). Limited separation was discernible in the targeted serum metabolome between baseline and post-intervention when projected on the first and second principal component(s). However, univariate testing identified 11 fatty acids that had lower concentrations (false discovery rate < 0.05) in post-intervention samples. CONCLUSIONS: These findings demonstrate that this short-term SIT intervention had limited effect on the serum metabolome at rest, but a subfraction of fatty acids are potentially sensitive to short-term exercise training.

Fasted Sprint Interval Training Results in Some Beneficial Skeletal Muscle Metabolic, but Similar Metabolomic and Performance Adaptations Compared With Carbohydrate-Fed Training in Recreationally Active Male.

Endurance training in fasted conditions (FAST) induces favorable skeletal muscle metabolic adaptations compared with carbohydrate feeding (CHO), manifesting in improved exercise performance over time. Sprint interval training (SIT) is a potent metabolic stimulus, however nutritional strategies to optimize adaptations to SIT are poorly characterized. Here we investigated the efficacy of FAST versus CHO SIT (4-6 × 30-s Wingate sprints interspersed with 4-min rest) on muscle metabolic, serum metabolome and exercise performance adaptations in a double-blind parallel group design in recreationally active males. Following acute SIT, we observed exercise-induced increases in pan-acetylation and several genes associated with mitochondrial biogenesis, fatty acid oxidation, and NAD+-biosynthesis, along with favorable regulation of PDK4 (p = .004), NAMPT (p = .0013), and NNMT (p = .001) in FAST. Following 3 weeks of SIT, NRF2 (p = .029) was favorably regulated in FAST, with augmented pan-acetylation in CHO but not FAST (p = .033). SIT induced increases in maximal citrate synthase activity were evident with no effect of nutrition, while 3-hydroxyacyl-CoA dehydrogenase activity did not change. Despite no difference in the overall serum metabolome, training-induced changes in C3:1 (p = .013) and C4:1 (p = .010) which increased in FAST, and C16:1 (p = .046) and glutamine (p = .021) which increased in CHO, were different between groups. Training-induced increases in anaerobic (p = .898) and aerobic power (p = .249) were not influenced by nutrition. These findings suggest some beneficial muscle metabolic adaptations are evident in FAST versus CHO SIT following acute exercise and 3 weeks of SIT. However, this stimulus did not manifest in differential exercise performance adaptations.

A Cell-Based Assessment of the Muscle Anabolic Potential of Blue Whiting (Micromesistius poutassou) Protein Hydrolysates.

Blue whiting (BW) represents an underutilised fish species containing a high-quality protein and amino acid (AA) profile with numerous potentially bioactive peptide sequences, making BW an economic and sustainable alternative source of protein. This study investigated the impact of three different BW protein hydrolysates (BWPH-X, Y and Z) on growth, proliferation and muscle protein synthesis (MPS) in skeletal muscle (C2C12) myotubes. BWPHs were hydrolysed using different enzymatic and heat exposures and underwent simulated gastrointestinal digestion (SGID), each resulting in a high degree of hydrolysis (33.41-37.29%) and high quantities of low molecular mass peptides (86.17-97.12% <1 kDa). C2C12 myotubes were treated with 1 mg protein equivalent/mL of SGID-BWPHs for 4 h. Muscle growth and myotube thickness were analysed using an xCelligence™ platform. Anabolic signalling (phosphorylation of mTOR, rpS6 and 4E-BP1) and MPS measured by puromycin incorporation were assessed using immunoblotting. BWPH-X significantly increased muscle growth (p < 0.01) and myotube thickness (p < 0.0001) compared to the negative control (amino acid and serum free media). Muscle protein synthesis (MPS), as measured by puromycin incorporation, was significantly higher after incubation with BWPH-X compared with the negative control, but did not significantly change in response to BWPH-Y and Z treatments. Taken together, these preliminary findings demonstrate the anabolic potential of some but not all BWPHs on muscle enhancement, thus providing justification for human dietary intervention studies to confirm and translate the results of such investigations to dietary recommendations and practices.

The Influence of Sitting, Standing, and Stepping Bouts on Cardiometabolic Health Markers in Older Adults.

Aside from total time spent in physical activity behaviors, how time is accumulated is important for health. This study examined associations between sitting, standing, and stepping bouts, with cardiometabolic health markers in older adults. Participants from the Mitchelstown Cohort Rescreen Study (N = 221) provided cross-sectional data on activity behaviors (assessed via an activPAL3 Micro) and cardiometabolic health. Bouts of ≥10-, ≥30-, and ≥60-min sitting, standing, and stepping were calculated. Linear regression models were fitted to examine the associations between bouts and cardiometabolic health markers. Sitting (≥10, ≥30, and ≥60 min) and standing (≥10 and ≥30 min) bouts were detrimentally associated with body composition measures, lipid markers, and fasting glucose. The effect for time spent in ≥60-min sitting and ≥30-min standing bouts was larger than shorter bouts. Fragmenting sitting with bouts of stepping may be targeted to benefit cardiometabolic health. Further insights for the role of standing need to be elicited.

Divergent serum metabolomic, skeletal muscle signaling, transcriptomic, and performance adaptations to fasted versus whey protein-fed sprint interval training.

Sprint interval training (SIT) is a time-efficient alternative to endurance exercise, conferring beneficial skeletal muscle metabolic adaptations. Current literature has investigated the nutritional regulation of acute and chronic exercise-induced metabolic adaptations in muscle following endurance exercise, principally comparing the impact of training in fasted and carbohydrate-fed (CHO) conditions. Alternative strategies such as exercising in low CHO, protein-fed conditions remain poorly characterized, specifically pertaining to adaptations associated with SIT. Thus, this study aimed to compare the metabolic and performance adaptations to acute and short-term SIT in the fasted state with preexercise hydrolyzed (WPH) or concentrated (WPC) whey protein supplementation. In healthy males, preexercise protein ingestion did not alter exercise-induced increases in PGC-1α, PDK4, SIRT1, and PPAR-δ mRNA expression following acute SIT. However, supplementation of WPH beneficially altered acute exercise-induced CD36 mRNA expression. Preexercise protein ingestion attenuated acute exercise-induced increases in muscle pan-acetylation and PARP1 protein content compared with fasted SIT. Acute serum metabolomic differences confirmed greater preexercise amino acid delivery in protein-fed compared with fasted conditions. Following 3 wk of SIT, training-induced increases in mitochondrial enzymatic activity and exercise performance were similar across nutritional groups. Interestingly, resting muscle acetylation status was downregulated in WPH conditions following training. Such findings suggest preexercise WPC and WPH ingestion positively influences metabolic adaptations to SIT compared with fasted training, resulting in either similar or enhanced performance adaptations. Future studies investigating nutritional modulation of metabolic adaptations to exercise are warranted to build upon these novel findings.NEW & NOTEWORTHY These are the first data to show the influence of preexercise protein on serum and skeletal muscle metabolic adaptations to acute and short-term sprint interval training (SIT). Preexercise whey protein concentrate (WPC) or hydrolysate (WPH) feeding acutely affected the serum metabolome, which differentially influenced acute and chronic changes in mitochondrial gene expression, intracellular signaling (acetylation and PARylation) resulting in either similar or enhanced performance outcomes when compared with fasted training.

Development of a multiplex assay to determine the expression of mitochondrial genes in human skeletal muscle.

NEW FINDINGS: What is the central question of this study? Can a custom-designed multiplex gene expression assay be used to quantify expression levels of a targeted group of mitochondrial genes in human skeletal muscle? What is the main finding and its importance? A custom-designed GeXP multiplex assay was developed, and the ability to accurately quantify expression of a targeted set of mitochondrial genes in human skeletal muscle was demonstrated. It holds distinct methodological and practical advantages over other commonly used quantification methods. ABSTRACT: Skeletal muscle is an important endocrine tissue demonstrating plasticity in response to external stimuli, including exercise and nutrition. Mitochondrial biogenesis is a common hallmark of adaptations to aerobic exercise training. Furthermore, altered expression of several genes implicated in the regulation of mitochondrial biogenesis, substrate oxidation and nicotinamide adenine dinucleotide (NAD+ ) biosynthesis following acute exercise underpins longer-term muscle metabolic adaptations. Gene expression is typically measured using real-time quantitative PCR platforms. However, interest has developed in the design of multiplex gene expression assays (GeXP) using the GenomeLab GeXP™ genetic analysis system, which can simultaneously quantify gene expression of multiple targets, holding distinct advantages in terms of throughput, limiting technical error, cost effectiveness, and quantifying gene co-expression. This study describes the development of a custom-designed GeXP assay incorporating the measurement of proposed regulators of mitochondrial biogenesis, substrate oxidation, and NAD+ biosynthetic capacity in human skeletal muscle and characterises the resting gene expression (overnight fasted and non-exercised) signature within a group of young, healthy, recreationally active males. The design of GeXP-based assays provides the capacity to more accurately characterise the regulation of a targeted group of genes with specific regulatory functions, a potentially advantageous development for future investigations of the regulation of muscle metabolism by exercise and/or nutrition.

Carbohydrate and Protein Co-Ingestion Postexercise Does Not Improve Next-Day Performance in Trained Cyclists.

Supplementing postexercise carbohydrate (CHO) intake with protein has been suggested to enhance recovery from endurance exercise. The aim of this study was to investigate whether adding protein to the recovery drink can improve 24-hr recovery when CHO intake is suboptimal. In a double-blind crossover design, 12 trained men performed three 2-day trials consisting of constant-load exercise to reduce glycogen on Day 1, followed by ingestion of a CHO drink (1.2 g·kg-1·2 hr-1) either without or with added whey protein concentrate (CHO + PRO) or whey protein hydrolysate (CHO + PROH) (0.3 g·kg-1·2 hr-1). Arterialized blood glucose and insulin responses were analyzed for 2 hr postingestion. Time-trial performance was measured the next day after another bout of glycogen-reducing exercise. The 30-min time-trial performance did not differ between the three trials (M ± SD, 401 ± 75, 411 ± 80, 404 ± 58 kJ in CHO, CHO + PRO, and CHO + PROH, respectively, p = .83). No significant differences were found in glucose disposal (area under the curve [AUC]) between the postexercise conditions (364 ± 107, 341 ± 76, and 330 ± 147, mmol·L-1·2 hr-1, respectively). Insulin AUC was lower in CHO (18.1 ± 7.7 nmol·L-1·2 hr-1) compared with CHO + PRO and CHO + PROH (24.6 ± 12.4 vs. 24.5 ± 10.6, p = .036 and .015). No difference in insulin AUC was found between CHO + PRO and CHO + PROH. Despite a higher acute insulin response, adding protein to a CHO-based recovery drink after a prolonged, high-intensity exercise bout did not change next-day exercise capacity when overall 24-hr macronutrient and caloric intake was controlled.

Motivational Impairment is Accompanied by Corticoaccumbal Dysfunction in the BACHD-Tg5 Rat Model of Huntington's Disease.

Neuropsychiatric symptoms, such as avolition, apathy, and anhedonia, precede the onset of debilitating motor symptoms in Huntington's disease (HD), and their development may give insight into early disease progression and treatment. However, the neuronal and circuit mechanisms of premanifest HD pathophysiology are not well-understood. Here, using a transgenic rat model expressing the full-length human mutant HD gene, we find early and profound deficits in reward motivation in the absence of gross motor abnormalities. These deficits are accompanied by significant and progressive dysfunction in corticostriatal processing and communication among brain areas critical for reward-driven behavior. Together, our results define early corticostriatal dysfunction as a possible pathogenic contributor to psychiatric disturbances and may help identify potential pharmacotherapeutic targets for the treatment of HD.

Regulation of GLUT4 translocation in an in vitro cell model using postprandial human serum ex vivo.

NEW FINDINGS: What is the research question? This study used a new experimental model, in which culture medium is conditioned with human serum ex vivo, to investigate nutrient-mediated regulation of GLUT4 translocation in skeletal muscle cells in vitro. What is the main finding and importance? Human serum stimulated GLUT4 translocation, an effect differentially modulated by whether the culture medium was conditioned with serum from fasted subjects or with serum collected after feeding of intact or hydrolysed whey protein. Conditioning cell culture medium with human serum ex vivo represents a new approach to elucidate the effects of ingesting specific nutrients on skeletal muscle cell metabolism. ABSTRACT: Individual amino acids, amino acid mixtures and protein hydrolysates stimulate glucose uptake in many experimental models. To replicate better in vitro the dynamic postprandial response to feeding in vivo, in the present study we investigated the effects of culture media conditioned with fasted and postprandial human serum on GLUT4 translocation in L6-GLUT4myc myotubes. Serum samples were collected from healthy male participants (n = 8) at baseline (T0), 60 (T60) and 120 min (T120) after the ingestion of 0.33 g (kg body mass)-1 of intact (WPC) or hydrolysed (WPH) whey protein and an isonitrogenous non-essential amino acid (NEAA) control. L6-GLUT4myc myotubes were starved of serum and amino acids for 1 h before incubation for 1 h in medium containing 1% postprandial human serum, after which GLUT4 translocation was determined via colorimetric assay. Medium conditioned with fasted human serum at concentrations of 5-20% increased cell surface GLUT4myc abundance. Incubation with serum collected after the ingestion of WPH increased cell surface GLUT4myc at T60 relative to T0 [mean (lower, upper 95% confidence interval)]; [1.13 (1.05, 1.22)], whereas WPC [0.98 (0.90, 1.07)] or NEAA [1.02 (0.94, 1.11)] did not. The differential increases in cell surface GLUT4myc abundance were not explained by differences in serum concentrations of total, essential and branched-chain amino acids or insulin, glucagon-like peptide 1 (GLP-1) and gastric inhibitory polypeptide (GIP). Using a new ex vivo, in vitro approach, cell culture medium conditioned with postprandial serum after the ingestion of a whey protein hydrolysate increased GLUT4 translocation in skeletal muscle cells.

Regulation of muscle protein synthesis in an in vitro cell model using ex vivo human serum.

NEW FINDINGS: What is the central question of this study? Can medium conditioned by ex vivo human serum regulate muscle protein synthesis in skeletal muscle cells in vitro? What is the main finding and its importance? This study demonstrates that medium conditioned by ex vivo human serum can regulate muscle protein synthesis in skeletal muscle cells in vitro via the mammalian Target of Rapomycin (mTOR) pathway, and this can be regulated differentially by fed and fasted ex vivo human serum. ABSTRACT: Human serum embodies the integrated systemic response to any condition or perturbation, which may regulate muscle protein synthesis (MPS). Conditioning of medium with human serum represents a physiologically relevant method of regulating MPS in vitro. The primary purpose of the study was the development of a model using ex vivo human serum to condition medium and regulate MPS in in vitro skeletal muscle cells. Four young healthy men reported to the laboratory after an overnight fast and were fed with 0.33 g (kg body mass)-1 whey protein. Blood samples were taken before (Fasted) and 60 min postprandial (Fed). Fully differentiated C2C12 skeletal muscle cells were nutrient and serum deprived for 1 h and subsequently treated with medium conditioned with Fasted or Fed ex vivo human serum (20%) for 4 h. The MPS was measured using the surface sensing of translation technique and activation of mTOR, P70S6K and 4EBP1 by Western blot. Fasted and fed ex vivo human serum increased MPS (P < 0.05). Although a strong effect (ƞ2  = 0.36) for increased MPS in Fed relative to Fasted was observed, this was not statistically significant (P > 0.05). Activation of mTOR, P70S6K and 4EBP1 was significantly increased after treatment with Fed compared with Fasted ex vivo human serum (P < 0.05). Here, we developed and optimized the conditions for culture of C2C12 skeletal muscle cells, measurement of MPS and signalling in medium conditioned by ex vivo human serum. Furthermore, the functionality of the model was demonstrated by comparison of the response to medium conditioned by Fasted and Fed ex vivo human serum.

Acute reduction of lower-body contractile function following a microbiopsy of m. vastus lateralis.

Twenty-three resistance trained men 18-35 years (23 [3] years, 1.8 [0.1] m, 81 [10] kg body mass, 2.3 [1.1] years resistance training experience; mean [SD]) performed repeated maximal voluntary isometric squats (ISQ) and countermovement jumps (CMJ) pre- and +30 minutes post a unilateral microbiopsy of m. vastus lateralis. ISQ and CMJ were simultaneously measured by two force plates sampling ipsilateral (biopsied) and contralateral (non-biopsied) limb force. Bilateral limb force (ipsilateral + contralateral) and imbalance (ipsilateral/bilateral) data are reported as % change from pre-biopsy (mean [95% CI]). A post-biopsy reduction in bilateral ISQ peak force (-17 [-23, -11] %; P < 0.001), ISQ rate of force development (RFD; -28 [-41, -15] %, P = 0.002) and CMJ peak take-off force (-7 [-13, -1]%, P = 0.019) occurred. Imbalance was observed for ISQ peak force (3.2 [2.1, 4.3] %, P < 0.001), RFD (2.8 [1.6, 4.0] %, P < 0.001) and CMJ landing (3.3 [1.0, 5.6] %, P = 0.009), resultant of a force transfer from the ipsilateral (biopsied) to the contralateral (non-biopsied) limb. These data suggest that in young, resistance trained men a modulatory influence on maximal voluntary static and dynamic lower-body contractile function is evoked acutely (+30 minutes) following a microbiopsy of m. vastus lateralis.

Sex-related differences in joint-angle-specific functional hamstring-to-quadriceps strength ratios.

PURPOSE: To examine and compare sex-related differences in the functioning of the hamstrings and quadriceps muscles and the isokinetic hamstrings eccentric-to-quadriceps concentric functional ratio (H/Q FUNC). METHODS: Fifty male and 46 female young adults completed this study. Each participant carried out an isokinetic assessment to determine isokinetic concentric and eccentric torques during knee extension and flexion actions at 3 different angular velocities (60, 180 and 300°/s) adopting a lying position. The H/Q FUNC was calculated using peak torque (PT) values and 3 different joint-angle-specific torque values (15°, 30° and 45° of knee extension). A repeated measures analysis of variance was used to compare the results, and post hoc analyses using Friedman correction were employed. RESULTS: There were statistically significant effects of angular velocity, joint angle and sex on the H/Q FUNC (p < 0.01). Thus, the H/Q FUNC ratio in both males and females decreases closer to full knee extension and with increasing movement velocity. The H/Q FUNC was also significantly lower in females compared to males, irrespective of moment velocity and joint angle. CONCLUSIONS: The findings of the current study reinforce the need to examine the H/Q FUNC ratio closer to full knee extension (where knee injury is most likely to occur) rather than using PT values which may not be as informative, as well as to focus preventive and rehabilitation training programmes on reducing quadriceps dominance by enhancing eccentric hamstring strength (especially in females who are at higher risk of injury). LEVEL OF EVIDENCE: III.

The Effect of Strength Training on Performance Indicators in Distance Runners.

Beattie, K, Carson, BP, Lyons, M, Rossiter, A, and Kenny, IC. The effect of strength training on performance indicators in distance runners. J Strength Cond Res 31(1): 9-23, 2017-Running economy (RE) and velocity at maximal oxygen uptake (VV[Combining Dot Above]O2max) are considered to be the best physiological performance indicators in elite distance runners. In addition to cardiovascular function, RE and VV[Combining Dot Above]O2max are partly dictated by neuromuscular factors. One technique to improve neuromuscular function in athletes is through strength training. The aim of this study was to investigate the effect of a 40-week strength training intervention on strength (maximal and reactive strength), VV[Combining Dot Above]O2max, economy, and body composition (body mass, fat, and lean mass) in competitive distance runners. Twenty competitive distance runners were divided into an intervention group (n = 11; 29.5 ± 10.0 years; 72.8 ± 6.6 kg; 1.83 ± 0.08 m) and a control group (n = 9; 27.4 ± 7.2 years; 70.2 ± 6.4 kg; 1.77 ± 0.04 m). During week 0, 20, and 40, each subject completed 3 assessments: physiology (V2 mmol·L BLa, V2 mmol·L BLa [blood lactate], V4 mmol·L BLa, RE, VV[Combining Dot Above]O2max, V[Combining Dot Above]O2max), strength (1 repetition maximum back squat; countermovement jump and 0.3 m drop jump), and body composition (body mass, fat mass, overall lean, and leg lean). The intervention group showed significant improvements in maximal and reactive strength qualities, RE, and VV[Combining Dot Above]O2max, at weeks 20 (p ≤ 0.05) and 40 (p ≤ 0.05). The control group showed no significant changes at either time point. There were no significant changes in body composition variables between or within groups. This study demonstrates that 40 weeks of strength training can significantly improve maximal and reactive strength qualities, RE, and VV[Combining Dot Above]O2max, without concomitant hypertrophy, in competitive distance runners.

An in vivo microdialysis characterization of the transient changes in the interstitial dialysate concentration of metabolites and cytokines in human skeletal muscle in response to insertion of a microdialysis probe.

Skeletal muscle has recently been described as an endocrine organ, capable of releasing cytokines and regulators of metabolism. Microdialysis of the interstitial space of skeletal muscle enables analysis of the release of such cytokines. The purpose of this study was to determine the transient changes in concentration of metabolites and cytokines in human skeletal muscle in a 7h period following the insertion of a microdialysis probe. In total, sixteen microdialysis catheters were inserted into the vastus lateralis of male participants (age 26.2±1.35y, height 180.8±3.89cm, mass 83.9±3.86kg, BMI 25.7±0.87kgm(-2), body fat 26.1±3.0%). Serial samples were analyzed by micro-enzymatic and multiplexed immunoassay. Muscle interstitial glucose and lactate levels remained stable throughout, amino acid concentrations stabilized after 2.5h, however, insertion of a microdialysis catheter induced a 29-fold increase in peak IL-6 (p<0.001) and 35-fold increase in peak IL-8 concentrations (p<0.001) above basal levels 6h post insertion. In contrast to stable amino acid, glucose and lactate concentrations after 2h, commonly reported markers of tissue homeostasis in in vivo microdialysis, the multi-fold increase in IL-6 and IL-8 following insertion of a microdialysis catheter is indicative of a sustained disturbance of tissue homeostasis.

Insulin signaling regulates fatty acid catabolism at the level of CoA activation.

The insulin/IGF signaling pathway is a highly conserved regulator of metabolism in flies and mammals, regulating multiple physiological functions including lipid metabolism. Although insulin signaling is known to regulate the activity of a number of enzymes in metabolic pathways, a comprehensive understanding of how the insulin signaling pathway regulates metabolic pathways is still lacking. Accepted knowledge suggests the key regulated step in triglyceride (TAG) catabolism is the release of fatty acids from TAG via the action of lipases. We show here that an additional, important regulated step is the activation of fatty acids for beta-oxidation via Acyl Co-A synthetases (ACS). We identify pudgy as an ACS that is transcriptionally regulated by direct FOXO action in Drosophila. Increasing or reducing pudgy expression in vivo causes a decrease or increase in organismal TAG levels respectively, indicating that pudgy expression levels are important for proper lipid homeostasis. We show that multiple ACSs are also transcriptionally regulated by insulin signaling in mammalian cells. In sum, we identify fatty acid activation onto CoA as an important, regulated step in triglyceride catabolism, and we identify a mechanistic link through which insulin regulates lipid homeostasis.

The impact of making-weight on cognitive performance in apprentice jockeys.

Jockeys regularly engage in rapid weight-loss practices in preparation for competition. These practices are thought to impair cognitive function, although the evidence in support of this theory remains inconclusive. The purpose of this study was to examine the effects of making-weight on cognitive function in apprentice jockeys in a simulated and competitive environment. Apprentice jockeys (n = 12) reduced their body mass by 4% in 48 h in a simulated environment using weight-loss methods typically adopted in preparation for racing. Simple and choice reaction time, attention, learning and memory were assessed before and after the weight loss. A further 10 apprentice jockeys performed the cognitive function assessment in a competitive racing environment at both a self-reported "normal" and "light" body mass. Hydration status and body mass were assessed in all trials. In the simulated environment, body mass was reduced by 4.2 ± 0.3%, yet no change in cognitive function was observed. Cognitive function also remained unchanged in the competitive environment after a body mass loss of 5.7 ± 1.9%. Typical reductions in body mass in preparation for racing have no effect on cognitive function in apprentice jockeys in a simulated and competitive environment. Further research is required to investigate the physiological mechanisms preventing the adverse effects of making-weight on cognitive function in jockeys.

Participatory Approaches in the Context of Research Into Workplace Health Promotion to Improve Physical Activity Levels and Reduce Sedentary Behavior Among Office-Based Workers: Scoping Review.

BACKGROUND: Participatory research (PR) involves engaging in cocreation with end users and relevant stakeholders throughout the research process, aiming to distribute power equitably between the end users and research team. Engagement and adherence in previous workplace health promotion (WHP) studies have been shown to be lacking. By implementing a PR approach, the insights of end users and stakeholders are sought in the co-design of feasible and acceptable intervention strategies, thereby increasing the relevance of the research. OBJECTIVE: This scoping review aims to explore, identify, and map PR techniques and their impact when used in office-based WHP interventions designed to improve physical activity (PA) or reduce sedentary behavior (SB). METHODS: The reporting of this scoping review followed the PRISMA-ScR (Preferred Reporting Items for Systematic reviews and Meta-Analyses extension for Scoping Reviews). A systematic literature search of 5 electronic databases-Web of Science, PubMed, Scopus, Google Scholar, and OpenGrey-was conducted, searching from January 1, 1995, to February 8, 2023. In total, 2 independent reviewers first screened the retrieved articles by title and abstract, and then assessed the full texts based on the inclusion and exclusion criteria. The search strategy and eligibility criteria were developed and guided by an a priori population (office-based working adults), intervention (a PA WHP intervention that took a PR approach), comparison (no comparison required), and outcome (PA or SB) framework. Data were charted and discussed via a narrative synthesis, and a thematic analysis was conducted. The included studies were evaluated regarding the degree of end user engagement throughout the research process and power shared by the researchers, using Arnstein's ladder of citizen participation. RESULTS: The search retrieved 376 records, of which 8 (2.1%) met the inclusion criteria. Four key strategies were identified: (1) end user focus groups, (2) management involvement, (3) researcher facilitators, and (4) workplace champions. The degree of engagement and power shared was relatively low, with 25% (2/8) of the studies determined to be nonparticipation studies, 25% (2/8) determined to be tokenistic, and 50% (4/8) determined to provide citizen power. CONCLUSIONS: This review provides a foundation of evidence on the current practices when taking a PR approach, highlighting that previous office-based PA WHP studies have been largely tokenistic or nonparticipative, and identified that the end user is only engaged with in the conception and implementation of the WHP studies. However, a positive improvement in PA and reduction in SB were observed in the included studies, which were largely attributed to implementing a PR approach and including the end user in the design of the WHP intervention. Future studies should aim to collaborate with workplaces, building capacity and empowering the workforce by providing citizen control and letting the end users "own" the research for a sustainable WHP intervention. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): RR2-10.1136/bmjopen-2021-054402.

Prevalence of reducing carbohydrate intake and fasted training in elite endurance athletes and association with bone injury.

There are conflicting reports both within the lay media and scientific literature regarding the use and benefit of dietary practices that aim to reduce CHO intake in endurance athletes. This study aimed to determine the prevalence of intentional reduction of CHO intake and fasted training in elite endurance-based athletes using a semi-quantitative questionnaire. Bone is a nutritionally modulated tissue; therefore, this study also aimed to explore if these dietary practices are potentially associated with bone injury incidence. The reported reduction of CHO intake was prevalent (28%) with the primary motivation being maintenance or manipulation of body composition. However, discrepancies in athletes' awareness of CHO intake were identified providing a potential avenue of intervention especially within applied practice. The use of fasted training was more prevalent (38%) with athletes using this practice for both body composition manipulation and promoting a desired adaptive response. Forty-four per cent of participants had suffered a radiographically confirmed bone injury at some point in their career. There was no association between reduction in CHO intake and bone injury incidence; however, the incidence of bone injury was 1.61 times higher in those who currently use fasted training compared to those who have never used it or who have used it in the past. Although a direct causal link between these dietary practices and the incidence of bone injury cannot be drawn, it provides robust justification for future investigations of the potential mechanisms that could explain this finding.