Analysis of epigenetic clocks links yoga, sleep, education, reduced meat intake, coffee, and a SOCS2 gene variant to slower epigenetic aging.

DNA methylation (DNAm) clocks hold promise for measuring biological age, useful for guiding clinical interventions and forensic identification. This study compared the commonly used DNAm clocks, using DNA methylation and SNP data generated from nearly 1000 human blood or buccal swab samples. We evaluated different preprocessing methods for age estimation, investigated the association of epigenetic age acceleration (EAA) with various lifestyle and sociodemographic factors, and undertook a series of novel genome-wide association analyses for different EAA measures to find associated genetic variants. Our results highlighted the Skin&Blood clock with ssNoob normalization as the most accurate predictor of chronological age. We provided novel evidence for an association between the practice of yoga and a reduction in the pace of aging (DunedinPACE). Increased sleep and physical activity were associated with lower mortality risk score (MRS) in our dataset. University degree, vegetable consumption, and coffee intake were associated with reduced levels of epigenetic aging, whereas smoking, higher BMI, meat consumption, and manual occupation correlated well with faster epigenetic aging, with FitAge, GrimAge, and DunedinPACE clocks showing the most robust associations. In addition, we found a novel association signal for SOCS2 rs73218878 (p = 2.87 × 10-8) and accelerated GrimAge. Our study emphasizes the importance of an optimized DNAm analysis workflow for accurate estimation of epigenetic age, which may influence downstream analyses. The results support the influence of genetic background on EAA. The associated SOCS2 is a member of the suppressor of cytokine signaling family known for its role in human longevity. The reported association between various risk factors and EAA has practical implications for the development of health programs to improve quality of life and reduce premature mortality associated with age-related diseases.

Elucidating the molecular mechanisms underlying the induction of autophagy by antidepressant-like substances in C57BL/6J mouse testis model upon LPS challenge.

The treatment of depression with pharmaceuticals is associated with many adverse side effects, including male fertility problems. The precise mechanisms by which these agents affect testicular cells remain largely unknown, but they are believed to induce cellular stress, which is sensed by the endoplasmic reticulum (ER) and the Golgi apparatus. These organelles are responsible for maintaining cellular homeostasis and regulating signal pathways that lead to autophagy or apoptosis. Therefore, in this study, we aimed to investigate the autophagy, ER, and Golgi stress-related pathways in mouse testis following treatment with antidepressant-like substances (ALS) and ALS combined with lipopolysaccharide (LPS). We found that most ALS and activated proteins are associated with the induction of apoptosis. However, when imipramine (IMI) was combined with NS-398 (a cyclooxygenase-2 inhibitor) after LPS administration, we observed a marked increase in the BECLIN1, Bcl-2, ATG16L, and LC3 expression, which are marker proteins of autophagosome formation. The expression of the BECN1 and ATG16L genes was also high compared to the control, indicating the induction of autophagy processes that may potentially protect mouse testicular cells from death and regulate metabolism in the testis. Our findings may provide a better understanding of the stress-related effects of specific ALS on the testis. Video Abstract.

Involvement of estrogen receptor alpha (ERα) and impairment of steroidogenesis after exposure to tris(2,3-dibromopropyl) isocyanurate (TBC) in mouse spermatogenic (GC-1 spg) cells in vitro.

Good-quality reproductive cells are essential for reproduction. Endocrine disruptors are widely available in the environment and are known to have an adverse effect on spermatogenesis and steroidogenesis. One of them is tris(2,3-dibromopropyl) isocyanurate (TBC), i.e. one of the novel brominated flame retardants (NBFR). TBC is a widely distributed ingredient used in the production of flame retardants. Currently, it is known to affect the hormonal system, but the exact mechanism of its action is unknown. Therefore, the aim of the study was to determine whether TBC alone and in cotreatment with BHPI (estrogen receptor alpha antagonist) has an impact on the expression of nuclear receptors involved in the formation of steroid hormones, proteins, and enzymes responsible for steroidogenesis and the levels of steroid hormones (E2, P4, and T) in the GC-1 spg cell line as a mouse model of spermatogenic cells in vitro. Our results indicate that ERα is involved in the mechanism of TBC action, while no activation of PPARγ, AhR, and IGF-1R was observed. In addition, a decrease in the levels of most of the analyzed proteins and enzymes involved in steroid conversion was observed. Only Cyp19a1 was upregulated after TBC, BHPI, and TBC with BHPI cotreatment. In all the analyzed groups, a significant decrease in P4 and a subtle decrease in T and E2 were observed in the production and secretion of the hormones to the culture medium, compared to the control. The obtained results confirm the involvement of TBC in the dysregulation of steroid biosynthesis, which may affect male fertility.

Acute Effect of Caffeine-Based Multi-Ingredient Supplement on Reactive Agility and Jump Height in Recreational Handball Players.

Pre-exercise caffeine and guarana-based multi-ingredient supplement (MS) consumption may be more effective for physical performance improvement than caffeine and guarana alone due to the synergistic effect of biologically active ingredients in multi-ingredient supplements. This study aimed to examine the acute effect of MS on the reactive agility and jump performance in recreational handball male players. A randomized, double-blind, crossover study involved twenty-four male handball players (body mass 74.6 ± 8.8 kg; body height 179 ± 7 cm; age 23.8 ± 1.4 years). Participants were tested under three conditions: placebo, caffeine + guarana (CAF + GUA), or MS ingestion 45 min before exercise tests. Participants performed a reactive agility test (Y-shaped test) and countermovement jump (CMJ). None of the supplements improved countermovement jump height (p = 0.06). The time needed to complete the agility test was significantly (p = 0.02) shorter in the MS condition than in the placebo. The differences in agility between PL vs. CAF + GUA and MS vs. CAF + GUA conditions were not statistically significant (p = 0.88 and p = 0.07, respectively). The results of this study indicate that the caffeine-based multi-ingredient performance was effective in improvement in reactive agility but not in jump height in recreational handball male players. A similar effect was not observed with CAF + GUA ingestion alone.

Beta-Alanine Supplementation and Sport Climbing Performance.

BACKGROUND: Supplementing ß-alanine (BA) improves exercise performance in efforts that are highly dependent on anaerobic glycolysis. As it has not yet been established whether it relates to climbing, the current study aimed to investigate the effects of BA on climbing-specific performance. METHODS: Fifteen elite climbers performed intermittent high-force high-velocity campus board exercise, and two bouldering traverses, hard and easy. They ingested 4.0 g·d-1 BA or placebo for four weeks in a double-blind, pre/post experimental design. RESULTS: In the campus board trial, ANOVA revealed a tendency toward significance (p = 0.066). Post hoc analysis revealed that there was a significant (p = 0.002) and "large" (d = 1.55) increase in the total number of "slaps" in the BA group. No significant supplementation × group interaction was found in "hard" traverse and a significant interaction for mean changes in number of moves (p = 0.025) and in time to failure (p = 0.044) on an "easy" traverse. Post hoc analysis revealed that only the BA group significantly improved from baseline in number of moves (+9.5) and time to failure (+32 s). Effect sizes were d = 1.73 and d = 1.44, respectively. CONCLUSIONS: Four weeks of BA supplementation can improve performance during continuous climbing lasting about 1 min and repeated bouts of upper body campus-like movements. However, it failed to enhance climbing of a shorter duration.

Effects of Co-Ingestion of ß-Hydroxy-ß-Methylbutyrate and L-Arginine α-Ketoglutarate on Jump Performance in Young Track and Field Athletes.

The aim of the study was to determine the effect of simultaneous supplementation of ß-hydroxy-ß-methylbutyrate and L-Arginine α-ketoglutarate on lower limb power and muscle damage in medium distance runners aged 15.3 (±0.9) years old. METHODS: The study group consisted of 40 volunteers aged 14-17 years practicing medium distance running for at least two years. The study lasted 12 days and followed a randomized, double-blind, placebo-controlled, parallel design. All subjects attended a familiarization session on day 0 before the test. The subjects were randomly divided into two groups: supplements and placebo group. The same training cycle protocol was used in both groups during the 12-day training period. Morning warm-up involved 10 min jogging at 60-75% of maximal heart rate and countermovement jump height measurement. Main training units were carried out for both groups with the same volume. Training load assessment (the daily session Rating of Perceived Exertion (s-RPE) method) method takes into consideration the intensity and the duration of the training session to calculate the "training load" (TL). RESULTS: At the end of the training cycle, a significant (p = 0.002) decrease in the countermovement jump (CMJ) height was found in the placebo group when compared to the baseline. In the supplement group, there was no decrease in the countermovement jump height. Creatine kinase and lactate dehydrogenase concentration increased during the training days similarly in both groups and decreased on rest days. There were no differences between groups in enzymes concentration. The research results indicate that the supplement combination used in the supplements group prevented a reduction in the CMJ values. In contrast to the supplements group, in the placebo group, the CMJ changes were statistically significant: a noticeable (p = 0.002) decrease in CMJ was noted between the baseline measurement and the 6th measurement. The well-being of the subjects from both groups changed significantly during the training period, and the intergroup differences in the mood level were similar and not statistically significant. CONCLUSIONS: The results of this study indicate that the daily co-supplementation with calcium salt of ß-hydroxy-ß-methylbutyrate (7.5 g) and L-Arginine α-ketoglutarate (10 g) during training might help to prevent decline in jump performance. No influence on muscle damage markers or mood was shown.

Effects of Pre-Workout Multi-Ingredient Supplement on Anaerobic Performance: Randomized Double-Blind Crossover Study.

BACKGROUND: The purpose of this research was to investigate the acute effects of a pre-workout supplement on anaerobic performance in resistance-trained men. METHODS: Twenty-three men underwent three randomized, double-blind testing sessions separated by a seven-day break. The participants performed three tests: isokinetic strength, three repetition maximum (3-RM) strength and Wingate. Statistical analysis was conducted in R environment. Linear mixed models were estimated via R package lme4. RESULTS: Mean T@0.2 s was significantly greater in supplemented condition for right and left knee flexors (PL: 103.2 ± 37.6 Nm; supplemented condition: 131.8 ± 29.3 Nm (p = 0.001)), and PL: 103.7 ± 39.3; supplemented condition: 129.4 ± 28.4 (p = 0.001)). T@0.2 s for right and left knee extensors (PL: 202.6 ± 58.6 Nm; supplemented condition: 237.2 ± 54.7 Nm (p = 0.001); PL: 203.3 ± 63.2 Nm, supplemented condition: 229.8 ± 50.8 Nm (p = 0.002)). Significant difference was in mean anaerobic power between supplemented and PL condition for right and left knee flexors (p = 0.002, p = 0.005) and for right and left knee extensors (p = 0.001 and p = 0.002). TTP was significantly shorter in supplemented condition for both sides knee flexors (p = 0.002). There was a significant difference for mean power in the Wingate test (placebo: 8.5 ± 0.6 W/kg; supplemented condition: 8.7 ± 0.5 W/kg (p = 0.038)). Mean 3-RM was significantly greater in supplemented condition (p = 0.001). CONCLUSIONS: The supplement significantly improves upper and lower body strength and power output in resistance-trained men.

Influence of Sunlight and Oral D3 Supplementation on Serum 25(OH)D Concentration and Exercise Performance in Elite Soccer Players.

The aim of this study was to evaluate the influence of natural sun exposure and six weeks of a high dose of vitamin D supplementation on vitamin D, testosterone and cortisol serum concentrations as well as speed, power and VO2max in professional soccer players. MATERIALS: The study was conducted from January to September. At the beginning of the study, 33 professional soccer players were enrolled; however, only 28 subjects (height 181.5 cm; body mass 77.81 ± 8.8 kg; body fat 12.38% ± 2.4% and muscle mass 40.27 ± 5.3 kg) completed the study. The research consisted of three stages. The first one, lasting 10 days, was conducted in January during a training camp in the south part of Cyprus at a latitude of 34 33°, where participants experienced natural sun exposure; it was called a winter sun exposure (WSE) period. The second stage, which was a supplementation period (SP), lasted 6 weeks, during which all subjects were randomly assigned either to an experimental group-EG (n = 15)-or a placebo group-PG (n = 13)-and were administered 6000 IU/d cholecalciferol or a placebo, respectively. The third stage took place in September, after summertime (summer sun exposure-SSE). The data of the 25(OH)D, free and total testosterone (fT, tT), cortisol as well as 5 and 30 m sprint tests (STs), power of the left leg (PLL) and VO2max were evaluated before and after the WSE period, the SP and SSE. RESULTS: In January, the baseline value of vitamin D in 12 subjects was ≤20 ng/mL, and 14 of them had levels between 20-30 ng/mL and 2 individuals >30 ng/mL. After the WSE period, significant changes in 25(OH)D, fT, tT and cortisol concentration, as well as in the 5 m ST, were observed. After the SP, in the EG, significant changes were found in 25(OH)D, fT, tT and the 5 m ST. Furthermore, a positive correlation between the concentration of 25(OH) fT and tT was observed. After SSE, 2 out of 28 players had <20 ng/mL 25(OH)D, 12 of them had 25(OH)D between 20 and 30 ng/mL and 14 of them had 25(OH)D between 30 and 50 ng/mL. Significant differences in 25(OH)D, fT, tT concentration and the 5 m ST performance were observed following SSE compared with the WSE period. CONCLUSION: Due to the serum level of 25(OH)D demonstrated by most participants at the beginning of the study and after summertime, all-year-round supplementation with high doses of vitamin D seems to be a reasonable solution to enhance high 25(OH)D concentration in blood and physical performance. In the middle of the winter, almost half of the soccer players were serum deficient of 25(OH)D. After ten days of sun exposure and 6 weeks of vitamin D supplementation, the concentration of 25(OH)D significantly increased, as did testosterone and results in the 5 m sprint test also improved. Therefore, athletes should be constantly monitored for serum levels of 25(OH)D throughout the year and should be supplemented if deficiencies or insufficient amounts of this vitamin occur.

Mechanism of Action and the Effect of Beta-Hydroxy-Beta-Methylbutyrate (HMB) Supplementation on Different Types of Physical Performance - A Systematic Review.

Beta-hydroxy-beta-methylbutyrate (HMB) has been used extensively as a dietary supplement for athletes and physically active people. HMB is a leucine metabolite, which is one of three branched chain amino acids. HMB plays multiple roles in the human body of which most important ones include protein metabolism, insulin activity and skeletal muscle hypertrophy. The ergogenic effects of HMB supplementation are related to the enhancement of sarcolemma integrity, inhibition of protein degradation (ubiquitin pathway), decreased cell apoptosis, increased protein synthesis (mTOR pathway), stimulation of the growth hormone/insulin-like growth factor-1 (GH/IGF-1) axis and enhancement of muscle stem cells proliferation and differentiation. HMB supplementation has been carried out with various groups of athletes. In endurance and martial arts athletes, HMB supplementation revealed positive effects on specific aerobic capacity variables. Positive results were also disclosed in resistance trained athletes, where changes in strength, body fat and muscle mass as well as anaerobic performance and power output were observed. The purpose of this review was to present the main mechanisms of HMB action, especially related to muscle protein synthesis and degradation, and ergogenic effects on different types of sports and physical activities.

The TFE-induced transient native-like structure of the intrinsically disordered σ47° domain of Escherichia coli RNA polymerase.

The transient folding of domain 4 of an E. coli RNA polymerase σ7° subunit (rECσ47°) induced by an increasing concentration of 2,2,2-trifluoroethanol (TFE) in an aqueous solution was monitored by means of CD and heteronuclear NMR spectroscopy. NMR data, collected at a 30% TFE, allowed the estimation of the population of a locally folded rECσ47° structure (CSI descriptors) and of local backbone dynamics ((15)N relaxation). The spontaneous organization of the helical regions of the initially unfolded protein into a TFE-induced 3D structure was revealed from structural constraints deduced from (15)N- to (13)C-edited NOESY spectra. In accordance with all the applied criteria, three highly populated α-helical regions, separated by much more flexible fragments, form a transient HLHTH motif resembling those found in PDB structures resolved for homologous proteins. All the data taken together demonstrate that TFE induces a transient native-like structure in the intrinsically disordered protein.

Backbone dynamics of TFE-induced native-like fold of region 4 of Escherichia coli RNA polymerase sigma70 subunit.

Folding of a recombinant protein rECsigma(70) (4) comprising domain 4 of E. coli RNA polymerase sigma(70) subunit, upon addition of 2,2,2-trifluoroethanol (TFE) to its aqueous solution, was monitored by heteronuclear NMR spectroscopy. The TFE-induced migration of resonance signals in a series of (15)N-HSQC spectra displayed sequence-dependent heterogeneity. A common trend of uniform upfield shift in both (1)H and (15)N dimensions, indicative of generation of helical structures, breaks down for some residues almost cooperatively at 10-15% TFE (v/v), pointing to the buildup of non-helical regions separating the initially induced helices. The preferences of residues to assume either helical or non-helical conformation are correlated with the location in the sequence rather than with their type. CSI descriptors and (15)N relaxation data obtained for the protein at 10% TFE allowed characterization of the stability of the pre-folded state of rECsigma(70) (4). By all the criteria applied, three highly populated alpha-helical regions separated by much more flexible residues forming a loop and a turn in the DNA-binding HLHTH motif were identified. The location of the secondary structure elements along the protein sequence coincides with those found in homologous proteins, and with the helix nucleation regions determined in unfolded rECsigma(70) (4) at low pH. The bimodal distribution of the (15)N relaxation parameters enabled identification of residues forming a framework of the folded protein strictly corresponding to the HLHTH motif, bracketed by unfolded terminal regions. Thus, in respect to rECsigma(70) (4) in aqueous solution TFE acts not only as a strong helix inducer, but also as a folding agent.