Protocol for a randomised controlled trial to investigate the effects of vitamin K2 on recovery from muscle-damaging resistance exercise in young and older adults-the TAKEOVER study.

BACKGROUND: Regular participation in resistance exercise is known to have broad-ranging health benefits and for this reason is prominent in the current physical activity guidelines. Recovery after such exercise is important for several populations across the age range and nutritional strategies to enhance recovery and modulate post-exercise physiological processes are widely studied, yet effective strategies remain elusive. Vitamin K2 supplementation has emerged as a potential candidate, and the aim of the current study, therefore, is to test the hypothesis that vitamin K2 supplementation can accelerate recovery, via modulation of the underlying physiological processes, following a bout of resistance exercise in young and older adults. METHODS: The current study is a two-arm randomised controlled trial which will be conducted in 80 (40 young (≤40 years) and 40 older (≥65 years)) adults to compare post-exercise recovery in those supplemented with vitamin K2 or placebo for a 12-week period. The primary outcome is muscle strength with secondary outcomes including pain-free range of motion, functional abilities, surface electromyography (sEMG) and markers of inflammation and oxidative stress. DISCUSSION: Ethical approval has been granted by the College of Medical Veterinary and Life Sciences Ethical Committee at the University of Glasgow (Project No 200190189) and recruitment is ongoing. Study findings will be disseminated through a presentation at scientific conferences and in scientific journals. TRIAL REGISTRATION: ClinicialTrials.gov NCT04676958. Prospectively registered on 21 December 2020.

A Phenotypic Approach for the Identification of New Molecules for Targeted Protein Degradation Applications.

Targeted protein degradation is an emerging new strategy for the modulation of intracellular protein levels with applications in chemical biology and drug discovery. One approach to enable this strategy is to redirect the ubiquitin-proteasome system to mark and degrade target proteins of interest (POIs) through the use of proteolysis targeting chimeras (PROTACs). Although great progress has been made in enabling PROTACs as a platform, there are still a limited number of E3 ligases that have been employed for PROTAC design. Herein we report a novel phenotypic screening approach for the identification of E3 ligase binders. The key concept underlying this approach is the high-throughput modification of screening compounds with a chloroalkane moiety to generate HaloPROTACs in situ, which were then evaluated for their ability to degrade a GFP-HaloTag fusion protein in a cellular context. As proof of concept, we demonstrated that we could generate and detect functional HaloPROTACs in situ, using a validated Von Hippel-Lindau (VHL) binder that successfully degraded the GFP-HaloTag fusion protein in living cells. We then used this method to prepare and screen a library of approximately 2000 prospective E3 ligase-recruiting molecules.

Exercise acutely increases vitamin D receptor expression in T lymphocytes in vitamin D-deficient men, independent of age.

NEW FINDINGS: What is the central question of this study? Does exercise affect vitamin D receptor expression in T lymphocytes in young, middle-aged and older adults? What is the main finding and its importance? Moderate-intensity cycling exercise increases vitamin D receptor expression in vitamin D-deficient men, independent of age, presenting a strategy to combat the prevalence of vitamin D deficiency. ABSTRACT: Vitamin D plays a key role in the modulation of the immune system, mediated through the intracellular vitamin D receptor (VDR). Exercise has been shown to influence the activity and availability of the VDR. The aim of this study was to investigate the effect of age on basal immune cell (T-lymphocyte) VDR expression and the subsequent effect of acute aerobic exercise to modulate VDR expression in peripheral T cells. Thirty-five men were included in the study (mean ± SD: age 44 ± 17 years and body mass index 25.7 ± 3.1 kg/m2 ), separated into three age groups: 18-30 (n = 12), 31-45 (n = 11) and 60-75 years (n = 12). Participants completed two trials [control (CON) and aerobic exercise (AE)], with blood samples collected pre- and postexercise (0, 1 and 3 h). Peripheral blood T cells were isolated and analysed for VDR expression by flow cytometry. The results show that advanced age is associated with lower VDR expression in T cells (882 ± 274, 796 ± 243 and 594 ± 174 geomean in the 18-30, 31-45 and 60-75 year age groups, respectively). Acute AE was successful at acutely increasing VDR expression in T cells, irrespective of age. Advanced age corresponds to a lower T-cell VDR expression, which might be responsible for age-associated development of chronic conditions and autoimmunity. Exercise was successful in increasing VDR expression in T cells irrespective of age and independent of exercise-induced T-cell mobilization.

Potential Cellular and Biochemical Mechanisms of Exercise and Physical Activity on the Ageing Process.

Exercise in young adults has been consistently shown to improve various aspects of physiological and psychological health but we are now realising the potential benefits of exercise with advancing age. Specifically, exercise improves cardiovascular, musculoskeletal, and metabolic health through reductions in oxidative stress, chronic low-grade inflammation and modulating cellular processes within a variety of tissues. In this this chapter we will discuss the effects of acute and chronic exercise on these processes and conditions in an ageing population, and how physical activity affects our vasculature, skeletal muscle function, our immune system, and cardiometabolic risk in older adults.

PROTAC-Mediated Degradation of Bruton's Tyrosine Kinase Is Inhibited by Covalent Binding.

The impact of covalent binding on PROTAC-mediated degradation of BTK was investigated through the preparation of both covalent binding and reversible binding PROTACs derived from the covalent BTK inhibitor ibrutinib. It was determined that a covalent binding PROTAC inhibited BTK degradation despite evidence of target engagement, while BTK degradation was observed with a reversible binding PROTAC. These observations were consistently found when PROTACs that were able to recruit either IAP or cereblon E3 ligases were employed. Proteomics analysis determined that the use of a covalently bound PROTAC did not result in the degradation of covalently bound targets, while degradation was observed for some reversibly bound targets. This observation highlights the importance of catalysis for successful PROTAC-mediated degradation and highlights a potential caveat for the use of covalent target binders in PROTAC design.

The combined effect of high-intensity intermittent training and vitamin D supplementation on glycemic control in overweight and obese adults.

High-intensity intermittent training (HIIT) has been shown to reduce the risk of chronic conditions including the development of type 2 diabetes mellitus (T2DM). Independently, a low vitamin D status has also been linked to the prevalence of T2DM. The aim of this study was to investigate if there was a synergistic metabolic effect of HIIT and vitamin D supplementation on glycemic control. A total of 20 male and female participants (age, 34 ± 9 year; BMI, 31.4 ± 2.8 kg·m-2 ) completed 6 weeks HIIT, and were randomized to ingest 100 µg·day-1 of vitamin D3 or placebo. Response to an oral glucose tolerance test (OGTT) was determined at baseline and at 72 h postintervention. Glucose tolerance was improved as a result of the HIIT intervention, shown through a reduction in glucose and insulin concentrations during the OGTT, accompanied by a decrease in glucose (829 ± 110 to 786 ± 139 mmol·h-1 ·L-1 ; P = 0.043) and insulin (8101 ± 4755-7024 ± 4489 mU·h-1 ·L-1 ; P = 0.049) area under the curve (AUC). Supplementation increased 25-hydroxyvitamin D3 concentration by 120% to a sufficiency status (P < 0.001). However, the consumption of vitamin D3 seemed to attenuate the glucose response during an OGTT. Triglyceride content was lowered following the intervention (P = 0.025). There was no effect of the intervention on insulin sensitivity (IS) indices: ISIMatsuda and HOMA-IR. Our findings demonstrate that HIIT improves glucose tolerance in nondiabetic overweight and obese adults; however vitamin D3 supplementation did not proffer any additional positive effects on the measured indices of metabolic health.

The Effect of a Single Bout of High Intensity Intermittent Exercise on Glucose Tolerance in Non-diabetic Older Adults.

Our aim was to investigate the acute effects of a single bout of high intensity intermittent training (HIIT) on glucose tolerance and other physiological and metabolic markers in non-diabetic older adults. Fourteen healthy older adults (age, 64 ± 2 y; BMI, 25.7 ± 2.8 kg·m-2) performed two acute exercise trials: continuous moderate intensity exercise (MOD) and HIIT, with the response to an oral glucose tolerance test (OGTT) determined <24 hours after. Inflammatory, haematological, and lipid parameters were also assessed the day after each trial. There was an effect of the trials on the insulin response to an OGTT (P=0.047), but not the glucose response. Following an acute bout of HIIT, insulin concentration during an OGTT was elevated at 60 min compared to the control trial (P=0.045), indicating more insulin was secreted, but glucose concentration was unchanged in all trials. The study findings demonstrate that a single bout of HIIT affects the insulin response but not the glycaemic response to a glucose load, proffering a potential benefit for metabolic health in older adults.