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Jakub Kortas,1 Ewa Ziemann,2 Jedrzej Antosiewicz3 1Department of Sport, Gdansk University of Physical Education and Sport, Gdansk, Poland; 2Department of Athletics, Strength and Conditioning, Poznan University of Physical Education, Poznan, Poland; 3Department of Bioenergetics and Physiology of Exercise, Medical University of Gdansk, Gdansk, PolandCorrespondence: Jakub KortasDepartment of Sport, Gdansk University of Physical Education and Sport, Kazimierza Górskiego 1, Gdansk 80-336, PolandEmail jakub.kortas@awfis.gda.plJedrzej AntosiewiczDepartment of Bioenergetics and Physiology of Exercise, Medical University of Gdansk, Marii Skłodowskiej-Curie 3a, Gdansk 80-210, PolandEmail jant@gumed.edu.plBackground: Excess iron accumulation in human tissue is associated with the diet, lack of exercise, or genetic factors. Iron accumulation increases the risk of acute myocardial infarction, diabetes, and cancer. On the other hand, exercise reduces the risk of several morbidities and influences iron metabolism. Here, we evaluated changes in iron metabolism induced by exercise in elderly women bearing the H63A HFE mutation.Purpose: To identify a factor that modulates the effect of exercise on iron metabolism. We investigated whether regular exercise induces similar changes in iron metabolism, mainly manifested by lowered body iron stores, in individuals bearing the wild-type (WT) and mutated HFE gene.Subjects and Methods: Seventy-six women (average age 69.2± 5.6 years old) were enrolled in the study. Thirty-nine women participated in 12 weeks of Nordic walking (NW) training; the remaining participants were assigned to the control group. Based on the H63A HFE mutation status, the NW group was divided into women bearing the mutation (HET, n=12) and women with the WT gene (WT, n=27).Results: The training resulted in a statistically significant reduction in the serum iron (p=0.03) and ferritin levels (p=0.001); hepcidin levels remained unchanged. No differences in these parameters were noted between the HET and WT groups.Conclusion: These observations suggest that a reduction in body iron stores might constitute an important aspect of the health-promoting effect of exercise, regardless of the genetic background.Keywords: physical activity, ferritin, inflammation, glucose

Jakub Kortas,1 Katarzyna Prusik,2 Damian Flis,3 Krzysztof Prusik,1 Ewa Ziemann,4 Neil Leaver,5 Jedrzej Antosiewicz6 1Department of Recreation and Tourism, Gdansk University of Physical Education and Sport, Gdansk, Poland; 2Department of Biomedical Basis of Health, Gdansk University of Physical Education and Sport, Gdansk, Poland; 3Department of Bioenergetics and Physiology of Exercise, Medical University of Gdansk, Gdansk, Poland; 4Department of Physiology and Pharmacology, Gdansk University of Physical Education and Sport, Gdansk, Poland; 5The Immunosuppression monitoring service (IMS) Laboratory, Royal Brompton & Harefield NHS Foundation Trust, Heart Science Centre, Harefield Hospital, Harefield, UK; 6Department of Biochemistry, Gdansk University of Physical Education and Sport, Gdansk, Poland Background: Despite several, well-documented pro-healthy effects of regular physical training, its influence on body iron stores in elderly people remains unknown. At the same time, body iron accumulation is associated with high risk of different morbidities.Purpose: We hypothesized that Nordic Walking training would result in pro-healthy changes in an elderly group of subjects by reducing body iron stores via shifts in iron metabolism-regulating proteins.Methods: Thirty-seven women aged 67.7±5.3 years participated in this study. They underwent 32 weeks of training, 1-hour sessions three times a week, between October 2012 and May 2013. Fitness level, blood morphology, CRP, vitamin D, ferritin, hepcidin, and soluble Hjv were assessed before and after the training.Results: The training program caused a significant decrease in ferritin, which serves as a good marker of body iron stores. Simultaneously, the physical cardiorespiratory fitness had improved. Furthermore, blood hepcidin was positively correlated with the ferritin concentration after the training. The concentration of blood CRP dropped, but the change was nonsignificant. The applied training resulted in a blood Hjv increase, which was inversely correlated with the vitamin D concentration.Conclusion: Overall the Nordic Walking training applied in elderly people significantly reduced blood ferritin concentration, which explains the observed decrease in body iron stores. Keywords: ferritin, hemojuvelin, hepcidin, vitamin D, anti-agingA Letter to the Editor has been recieved and published for this article.

There is general agreement that oxidative stress may induce apoptotic and necrotic cell death. Recently it has been shown that NADH can be considered an important antioxidant as it reacts with peroxyl and alkoxyl radicals under in vitro conditions. Therefore, in the present study we hypothesized that an increase in intracellular NADH using specific substrates will protect RL-34 cells against cytotoxicity of 2'-azobis (2-amidinopropane) dihydrochloride (AAPH), which is a peroxyl radical generating compound. Cells treated for 24 hours with 6.0 mM AAPH were severely damaged: mitochondria were vacuolated, and the level of free radicals significantly increased. Both apoptotic and necrotic cells were detected (11.1% and 11.4%, respectively) even after 5 hours of treatment. Pretreatment of the cells with substrates which increase the intracellular level of NADH, such as lactate, beta-hydroxybutyrate, and ethanol, distinctly inhibited AAPH-induced reactive oxygen species (ROS) formation and cell death. On the other hand, acetoacetate (AcA), which decrease the intracellular level of NADH, had opposite effects. Interestingly, NADH-generating substrates augment, while AcA reduced superoxide radical formation induced by AAPH. These results may suggest that although NADH generating substrates may exert some deleterious effects within a cell by inducing reductive stress, they diminish alkoxyl or peroxyl radical cytotoxicity. The protection is associated with a decrease in ROS formation measured by dichlorofluorescein, but with an increase in superoxide radical formation. [ABSTRACT FROM AUTHOR]

Purpose: Tryptophan (TRP) degradation through the kynurenine pathway is responsible for converting 95% of free TRP into kynurenines, which modulate skeletal muscle bioenergetics, immune and central nervous system activity. Therefore, changes in the kynurenines during exercise have been widely studied but not in the context of the effects of remote ischemic preconditioning (RIPC). In this study, we analyzed the effect of 14-day RIPC training on kynurenines and TRP in runners after running intervals of 20 × 400 m., Methods: In this study, 27 semi-professional long-distance runners were assigned to two groups: a RIPC group performing 14 days of RIPC training (n = 12), and a placebo group, SHAM (n = 15). Blood was collected for analysis before, immediately after, and at 6 h and 24 h after the run., Results: After the 14-day RIPC/SHAM intervention, post hoc analysis showed a significantly lower concentration of XANA and kynurenic acid to kynurenine ratio (KYNA/KYN) in the RIPC group than in the SHAM group immediately after the running test. Conversely, the decrease in serum TRP levels was higher in the RIPC population., Conclusion: RIPC modulates post-exercise changes in XANA and TRP levels, which can affect brain health, yet further research is needed., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Brzezińska, Mieszkowski, Stankiewicz, Kowalik, Reczkowicz, Niespodziński, Durzyńska, Kowalski, Borkowska, Antosiewicz and Kochanowicz.)

Purpose: Remote ischemic preconditioning (RIPC) is a method of protection against induced ischemia reperfusion injury, and an increasing number of studies showed some of its inconclusive ergogenic effects in sports. RIPC involves short cycles of cuff inflation followed by its deflation which may affect many body systems. While most of the studies focus on single RIPC effects, there is insufficient data regarding training-like repeated RIPC interventions. Thus, in this study, we analyzed the effect of a single- and consecutive 10-day RIPC procedure on a single leg, focusing on the exerkine levels and changes in inflammation markers following the Wingate Anaerobic Test (WAnT)., Methods: Two single-blinded, sham-controlled protocols were designed to evaluate the 1) single (crossover study) and 2) consecutive 10-day (parallel study) RIPC effects on the WAnT performance and exercise-induced lactate, glucose, exerkine, and inflammation markers (BDNF; IL-6; IL-10; IL-15; LIF; oncostatin M). In each protocol, 37 physically active men (19.98 ± 1.17 years) were randomly assigned into two groups according to a particular study design., Results: An increase in participants' mean (4.81%, p < 0.05) and peak power (6.25%, p < 0.05) during the WAnT was observed only after the consecutive 10-day RIPC. Similarly, a significant 15.5% (p < 0.05) decrease in the IL-6 concentration 120 min after the WAnT was observed only in the consecutive 10-day RIPC protocol, as well as a 12.2% (p < 0.01) increase in oncostatin M 60 min after the WAnT., Conclusion: The results demonstrate the efficacy of the consecutive 10-day RIPC procedure in modulating exercise performance and post-exercise inflammation markers., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kochanowicz, Brzezinska, Mieszkowski, Kochanowicz, Niespodzinski, Surmiak, Reczkowicz, Borkowska and Antosiewicz.)

Background: Increased inflammation and heightened physiological stress reactivity have been associated with pathophysiology of depressive symptoms. The underlying biological mechanisms by which inflammation and stress may influence neurogenesis are changes in the kynurenine (KYN) pathway, which is activated under stress. Supplementation with n -3 polyunsaturated fatty acids ( n -3 PUFAs) has anti-inflammatory properties and can increase stress resilience. Whether n -3 PUFAs alter KYN stress response is unknown. Objectives: This placebo-controlled study investigated the effect of n -3 PUFAs on KYN metabolism, inflammation, depressive symptoms, and mood. Moreover, stress-induced changes following a laboratory stressor have been assessed. Methods: In this placebo-controlled study, 47 healthy male adults received either 4 g n -3 PUFAs per day (Omega-3 group) or a placebo (Placebo group) for 12 weeks. Results: A significant group-by-time interaction was found for the inflammatory markers gp130 (F = 7.07, p = 0.011), IL-6R alpha (F = 10.33, p = 0.003), and TNF&#95;RI (F= 10.92, p = 0.002). No significant group-by-time interactions were found for KYN metabolites, depressive symptoms, and mood (except for Hedonic tone (F = 6.50, p = 0.014)), nor for stress-induced changes in KYN metabolites and mood following a laboratory stressor. Conclusions: Overall, increased n -3 PUFA levels in healthy men ameliorate inflammatory markers but do not ameliorate KYN metabolism, depressive symptoms, mood, or KYN metabolism and mood following a stress induction. This study was registered at ClinicalTrials.gov with the identifier NCT05520437 (30/08/2022 first trial registration).

Exercise-induced inflammation can influence iron metabolism. Conversely, the effects of vitamin D 3 , which possesses anti-inflammatory properties, on ultramarathon-induced heart damage and changes in iron metabolism have not been investigated. Thirty-five healthy long-distance semi-amateur runners were divided into two groups: one group received 150,000 IU of vitamin D 3 24 h prior to a race ( n = 16), while the other group received a placebo ( n = 19). Serum iron, hepcidin (HPC), ferritin (FER), erythroferrone (ERFE), erythropoietin (EPO), neopterin (NPT), and cardiac troponin T (cTnT) levels were assessed. A considerable effect of ultramarathon running on all examined biochemical markers was observed, with a significant rise in serum levels of ERFE, EPO, HPC, NPT, and cTnT detected immediately post-race, irrespective of the group factor. Vitamin D 3 supplementation showed a notable interaction with the UM, specifically in EPO and cTnT, with no other additional changes in the other analysed markers. In addition to the correlation between baseline FER and post-run ERFE, HPC was modified by vitamin D. The ultramarathon significantly influenced the EPO/ERFE/HPC axis; however, a single substantial dose of vitamin D 3 had an effect only on EPO, which was associated with the lower heart damage marker cTnT after the run.

Introduction: Artistic gymnastics is one of the most demanding sports disciplines, with the athletes demonstrating extremely high levels of explosive power and strength. Currently, knowledge of the effect of gymnastic training adaptation on exercise-induced inflammatory response is limited. The study aimed to evaluate inflammatory response following lower- and upper-body high-intensity exercise in relation to the iron status in gymnasts and non-athletes. Methods: Fourteen elite male artistic gymnasts (EAG, 20.6 ± 3.3 years old) and 14 physically active men (PAM, 19.9 ± 1.0 years old) participated in the study. Venous blood samples were taken before and 5 min and 60 min after two variants of Wingate anaerobic test (WAnT), upper-body and lower-body WAnT. Basal iron metabolism (serum iron and ferritin) and acute responses of selected inflammatory response markers [interleukin (IL) 6, IL-10, and tumour necrosis factor α] were analysed. Results: EAG performed significantly better during upper-body WAnT than PAM regarding relative mean and peak power. The increase in IL-6 levels after upper-body WAnT was higher in EAG than in PAM; the opposite was observed after lower-body WAnT. IL-10 levels were higher in EAG than in PAM, and tumour necrosis factor α levels were higher in PAM than those in EAG only after lower-body WAnT. The changes in IL-10 correlated with baseline serum iron and ferritin in PAM. Discussion: Overall, gymnastic training is associated with the attenuation of iron-dependent post-exercise anti-inflammatory cytokine secretion., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Kochanowicz, Waldziński, Niespodziński, Brzezińska, Kochanowicz, Antosiewicz and Mieszkowski.)

Researchers have studied the effects of exercise on serum methyl-arginine and vitamin D metabolites; however, the effects of exercise combined with antioxidants are not well documented. Since oxidative stress affects the metabolism of vitamin D and methyl-arginine, we hypothesised that the antioxidant coenzyme Q 10 (CoQ 10 ) might modulate exercise-induced changes. A group of twenty-eight healthy men participated in this study and were divided into two groups: an experimental group and a control group. The exercise test was performed until exhaustion, with gradually increasing intensity, before and after the 21-day CoQ 10 supplementation. Blood samples were collected before, immediately after, and 3 and 24 h after exercise. CoQ 10 , vitamin D metabolites, asymmetric dimethylarginine (ADMA), symmetric dimethylarginine, methylarginine, dimethylamine, arginine, citrulline, and ornithine were analysed in serum samples. CoQ 10 supplementation caused a 2.76-fold increase in the concentration of serum CoQ 10 . Conversely, the 25(OH)D 3 concentration increased after exercise only in the placebo group. ADMA increased after exercise before supplementation, but a decrease was observed in the CoQ 10 supplementation group 24 h after exercise. In conclusion, our data indicate that CoQ 10 supplementation modifies the effects of exercise on vitamin D and methyl-arginine metabolism, suggesting its beneficial effects. These findings contribute to the understanding of how antioxidants like CoQ 10 can modulate biochemical responses to exercise, potentially offering new insights for enhancing athletic performance and recovery.

Introduction: Tryptophan's (Trp) metabolites are undervalued markers of human health. Their serum concentrations are modified by physical exercise and other factors, among which fasting has a well-documented role. Although this mechanism is hardly explored, thus, the study aimed to determine the effect of the 8-day fasting period and the impact of such a procedure on a single bout of an endurance exercise on the concentration of kynurenine pathway (KP) metabolites., Methods: 10 participants fasted for 8 days, and 10 as a control group participated in the study. The exercise was performed at baseline after an overnight fast and repeated post 8 days., Results: The 8 days of fasting increased the resting 3-hydroxy-L-kynurenine (3HK), picolinic acid (PA), kynurenic acid (KYNA), and xanthurenic acid (XA) serum concentration. Also elevated phenylalanine (Phe) and tyrosine (Tyr) levels were recorded, suggesting expanded proteolysis of muscle proteins. In turn, physical activity caused a decrease in the concentration of 3-hydroxyanthranilic acid (3HAA) and PA after fasting. The obtained results were not recorded in controls., Conclusion: The results of this study show that the health-promoting effects of fasting are associated with changes in the KYN pathway. The increase in the concentration of PA and XA metabolites following fasting is capable of penetrating the blood-brain barrier, and KYNA, which initiates several beneficial changes, supports this assumption., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Juhas, Reczkowicz, Kortas, Żychowska, Pilis, Ziemann, Cytrych, Antosiewicz and Borkowska.)

Background and Aims: This study evaluated whether stored iron determines the adaptive response induced by Nordic walking (NW) training combined with 10 hours' time-restricted eating (TRE) in older adults., Trial Design and Methods: Twenty-four participants underwent 12-week NW training supported by 10 h of TRE. The group was divided due to baseline ferritin concentration low < 75 ng/ml (LF) and high level ≥ 75 ng/ml (HF). Body composition, physical fitness and blood collection were assessed at baseline and post-intervention., Results: NW + TRE induced a statistically significant decrease in ferritin levels in all participants (p = 0.01). Additionally, statistically significant intergroup differences in the LF vs. HF in the reduction of serum ferritin levels (p = 0.04) were observed. The procedure NW + TRE diminished HbA1c levels (p < 0.01) and glucose in all participants (p = 0.05). The range of HbA1c drop was more pronounced among those participants who experienced a greater decrease in the stored iron (p = 0.04, [Formula: see text]=0.17, F=4.59). Greater changes in body weight and percent of body fat were recorded in the HF group (for both p<0.01)., Conclusion: Body iron stores determine the effects of a 12-week NW + TRE intervention on serum ferritin. The changes in HbA1c are more pronounced in subjects with a higher decrease in serum ferritin., Trial Registration: All experimental protocols were approved by the Bioethical Committee of the Regional Medical Society in Gdansk, Poland (NKBBN/330/2021) according to the Declaration of Helsinki. We confirm that all methods were carried out in accordance with relevant guidelines and regulations. The trial was registered as a clinical trial (NCT05229835, date of first registration: 14/01/2022, direct link: https://classic., Clinicaltrials: gov/ct2/show/NCT05229835 ). Informed consent was obtained from all subjects., (© 2024. The Author(s).)

Background: The prevalence of depressive symptoms (DS) among physically active individuals tends to be lower compared to sedentary controls. This association seems to be moderated by gender and level of physical activity (PA). The objective of this meta-analysis was to assess the relationship between PA and DS in males based on different levels of PA., Methods: A systematic review and meta-analyses were conducted following the PRISMA Statement for Systematic Reviews. The literature search was conducted from January 1, 2003 to February 20, 2023. Cross-sectional and cohort studies including male participants aged 18 years or older were included in the analysis. Evidence from selected studies was synthesised as differences between odds ratios to assess whether DS were exhibited among those who were engaged in low, moderate, and high PA via random-effects meta-analyses. This study is registered on PROSPERO, number CRD42023417219., Results: Out of 1737 records identified, 5 eligible studies were included with a total of 35,811 participants. Results indicated significant effects of moderate PA on DS (OR = 0.68; 95 % C.I. 0.50-0.93). No effect of low and high PA on DS was found (OR = 0.79; 95 % C.I. 0.52-1.20 and OR = 0.78; 95 % C.I. 0.47-1.30)., Conclusion: Males who engage in moderate PA present lower prevalence of DS compared to no-PA reference. Such associations were not found for low or high PA. Hence, mental health benefits of PA could possibly be achieved at appropriate levels of PA. High heterogeneity between the studies should be considered when interpreting the results., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

Swim training has increased the life span of the transgenic animal model of amyotrophic lateral sclerosis (ALS). Conversely, the progress of the disease is associated with the impairment of iron metabolism and insulin signaling. We used transgenic hmSOD1 G93A (ALS model) and non-transgenic mice in the present study. The study was performed on the muscles taken from trained (ONSET and TERMINAL) and untrained animals at three stages of the disease: BEFORE, ONSET, and TERMINAL. In order to study the molecular mechanism of changes in iron metabolism, we used SH-SY5Y and C2C12 cell lines expression vector pcDNA3.1 and transiently transfected with specific siRNAs. The progress of ALS resulted in decreased P-Akt/Akt ratio, which is associated with increased proteins responsible for iron storage ferritin L, ferritin H, PCBP1, and skeletal muscle iron at ONSET. Conversely, proteins responsible for iron export- TAU significantly decrease. The training partially reverses changes in proteins responsible for iron metabolism. AKT silencing in the SH-SY5Y cell line decreased PCBP2 and ferroportin and increased ferritin L, H, PCBP1, TAU, transferrin receptor 1, and APP. Moreover, silencing APP led to an increase in ferritin L and H. Our data suggest that swim training in the mice ALS model is associated with significant changes in iron metabolism related to AKT activity. Down-regulation of AKT mainly upregulates proteins involved in iron import and storage but decreases proteins involved in iron export., Competing Interests: Declaration of competing interest The authors have no relevant financial or non-financial interests to disclose., (Copyright © 2023. Published by Elsevier B.V.)

The effects of long-term omega-3 polyunsaturated fatty acid (n-3 PUFA) supplementation during endurance training on tryptophan (Trp) metabolism and mental state of healthy individuals have not been evaluated so far. Concentrations of plasma Trp, its metabolites and IL-6 were assessed in 26 male runners before and after a 12-week training program combined with supplementation of n-3 PUFAs (O-3 + TRAIN group) or medium chain triglycerides (MCTs; TRAIN group). After the 12-week program participants' mood before and after stress induction was also assessed. The effects of the same supplementation protocol were evaluated also in 14 inactive subjects (O-3 + SEDEN group). Concentrations of 3-hydroxykynurenine (3-HK) and picolinic acid (PA) significantly increased only in the O-3 + TRAIN group (p = 0.01; [Formula: see text] = 0.22 and p = 0.01; [Formula: see text]= 0.26). Favorable, but not statistically significant changes in the concentrations of kynurenic acid (KYNA) (p = 0.06; [Formula: see text]= 0.14), xanthurenic acid (XA) (p = 0.07; [Formula: see text]= 0.13) and 3-hydroxyanthranilic acid (3-HAA) (p = 0.06; [Formula: see text]= 0.15) and in the ratio of neurotoxic to neuroprotective metabolites were seen also only in the O-3 + TRAIN group. No changes in mood and IL-6 concentrations were observed in either group. Supplementation with n-3 PUFAs during endurance training has beneficial effects on Trp's neuroprotective metabolites.Trial registry: This study was registered at ClinicalTrials.gov with identifier NCT05520437 (14/07/2021 first trial registration and 2018/31/N/NZ7/02962 second trial registration)., (© 2024. The Author(s).)

Multiple myeloma (MM) is an incurable hematologic malignancy originating from clonal plasma cell proliferation within the bone marrow, predominantly affecting older individuals. While anemia serves as a diagnostic criterion for MM, it often ameliorates upon achieving disease remission. Iron metabolism parameters have emerged as potential prognostic indicators in MM. Notably, physical exercise has been established to influence iron metabolism. This study aimed to assess alterations in serum iron, ferritin, and transferrin concentrations, as well as leukocyte gene expression, in MM patients undergoing a six-week cycle of Nordic walking training. Thirty patients divided into an exercise group (NW, n = 15, mean age 63.1 ± 8.4 years) and a control group (CG, n = 15, mean age: 63.5 ± 3.6 years) completed the study protocol. Blood samples were collected at baseline, after three and six weeks of training, and after nine weeks. Serum ferritin, transferrin, and iron concentrations were measured, along with the leukocyte expression of genes. Additionally, serum oxidative damage marker levels were determined. Following the Nordic walking training cycle, a declining trend in serum ferritin concentrations was observed. Intracellular mRNA levels of genes associated with iron metabolism were positively influenced by the training regimen, indicating the potential impact of this physical activity on gene expression and ferritin concentrations. Although positive trends were noted, extended training periods might be requisite for significant changes. To conclude, moderate-intensity exercise induces favorable shifts in the analyzed parameters among MM patients, potentially influencing disease progression. Consequently, Nordic walking training is a safe recommendation for MM patients, though sustained training beyond six weeks could be necessary for notable effects on iron metabolism factors.

Introduction: Although impacts of physical activity on cognitive functions have been intensively investigated, they are still far from being completely understood. The aim of this study was to evaluate the effect of 12 weeks of the Nordic Walking training with BungyPump resistance poles (NW-RSA) on the amino acid and kynurenine profiles as well as selected myokine/exerkine concentrations, which may modify the interface between physical and cognitive functions., Methods: A group of 32 older adults participated in the study. Before and after the intervention, body composition, cognitive functions, and physical performance were assessed. Blood samples were taken before and 1 h after the first and last sessions of the NW-RSA training, to determine circulating levels of exercise-induced proteins, i.e., brain-derived neurotrophic factor (BDNF), irisin, kynurenine (KYN), metabolites, and amino acids., Results: The NW-RSA training induced a significant improvement in cognitive functions and physical performance as well as a reduction in fat mass (p = 0.05). Changes were accompanied by a decline in resting serum BDNF (p = 0.02) and a slight reduction in irisin concentration (p = 0.08). Still, changes in irisin concentration immediately after the NW-RSA intervention depended on shifts in kynurenine-irisin dropped as kynurenine increased. The kynurenine-to-tryptophan and phenylalanine-to-tyrosine ratios decreased significantly, suggesting their possible involvement in the amelioration of cognitive functions. No changes of glucose homeostasis or lipid profile were found. Shifts in the concentrations of selected amino acids might have covered the increased energy demand in response to the NW-RSA training and contributed to an improvement of physical performance., Conclusion: Regular Nordic Walking training with additional resistance (BungyPump) improved cognitive functions and physical performance. These positive effects were associated with a reduced BDNF concentration and kynurenine-to-tryptophan ratio as well as changes in the amino acid profile., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Rodziewicz-Flis, Juhas, Kortas, Jaworska, Bidzan-Bluma, Babińska, Micielska, Żychowska, Lombardi, Antosiewicz and Ziemann.)

Exercise affects serum levels of amino acids and their metabolites, with important metabolic consequences. Since vitamin D impacts skeletal muscle protein degradation, we hypothesised that it would also impact exercise-induced changes in serum amino acid levels and the serum levels of arginine metabolites, influencing the body's ability to synthesise NO. Accordingly, we analysed the effect of a single high-dose vitamin D supplementation on the serum levels of various amino acids in ultramarathon runners. Thirty-five male amateur runners were assigned to the supplemented group, administered 150,000 IU vitamin D in vegetable oil 24 h before the run ( n = 16), or the control (placebo) group ( n = 19). Blood was sampled 24 h before, immediately after, and 24 h after the run. Changes in the serum levels of some amino acids were distinct in the two groups. The asymmetric dimethyl arginine levels were significantly decreased immediately after the run and increased 24 h later and were not affected by the supplementation. The symmetric dimethyl arginine levels were increased after the run in both groups but were lower in the supplemented group than in the placebo group 24 h after the run. The dimethylamine levels increased significantly in the supplemented group as compared to the placebo group. In conclusion, vitamin D impacts exercise-induced changes in serum amino acids and methylated arginine metabolites.

Changes in serum concentration of methylarginines and amino acids after exercise are well documented, whereas the effects of exercise applied together with fasting are still debated and not thoroughly studied. Thus, we hypothesised that alterations in methylarginines such as ADMA, SDMA and L-NMMA might be responsible for decreased exercise performance after 8 days of fasting. Additionally, we propose that conditions in which the human body is exposed to prolonged fasting for more than a week elicit a distinctly different response to exercise than after overnight fasting. A group of 10 healthy men with previous fasting experience participated in the study. The exercise test was performed until exhaustion with a gradually increasing intensity before and after the 8-day fast. Blood samples were collected before and immediately after exercise. ADMA, SDMA, L-NMMA, dimethylamine and amino acids were analysed in serum samples by ID-LC-MS/MS. SDMA, L-NMMA and dimethylamine significantly decreased after 8 days of fasting, whereas ADMA did not change. BCAA, Phe, alanine and some other amino acids increased after fasting. Exercise-induced changes in amino acids were distinct after an 8-day fast compared to overnight fasting. A decrease in physical performance accompanied all of these alterations. In conclusion, our data indicate that neither methyl-arginine changes nor the Trp/BCAA ratio can explain exercise-induced fatigue after fasting. However, the observed decrease in hArg concentration suggests the limited synthesis of creatine, possibly contributing to reduced physical performance.

The quantitative polymerase chain reaction (qRT-PCR) technique gives promising opportunities to detect and quantify RNA targets and is commonly used in many research fields. This study aimed to identify suitable reference genes for physical exercise and omega-3 fatty acids supplementation intervention. Forty healthy, physically active men were exposed to a 12-week eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) supplementation and standardized endurance training protocol. Blood samples were collected before and after the intervention and mRNA levels of six potential reference genes were tested in the leukocytes of 18 eligible participants using the qRT-PCR method: GAPDH (Glyceraldehyde-3-phosphate dehydrogenase), ACTB ( Beta actin ), TUBB ( Tubulin Beta Class I ), RPS18 ( Ribosomal Protein S18 ), UBE2D2 ( Ubiquitin-conjugating enzyme E2 D2 ), and HPRT1 ( Hypoxanthine Phosphoribosyltransferase 1 ). The raw quantification cycle (Cq) values were then analyzed using RefFinder, an online tool that incorporates four different algorithms: NormFinder, geNorm, BestKeeper, and the comparative delta-Ct method. Delta-Ct, NormFinder, BestKeeper, and RefFinder comprehensive ranking have found GAPDH to be the most stably expressed gene. geNorm has identified TUBB and HPRT as the most stable genes. All algorithms have found ACTB to be the least stably expressed gene. A combination of the three most stably expressed genes, namely GAPDH, TUBB, and HPRT, is suggested for obtaining the most reliable results.

The use of herbal medicinal products and supplements in amateur and professional sports has increased in the last decades. This is because most of these products and supplements contain bioactive compounds with a variety of biological properties that exert a physiological effect on the human body. The aim of this study was to analyze the effect of dietary supplementation with lyophilized black chokeberry extract on the levels of pro-inflammatory cytokines, hepcidin, and selected markers of iron metabolism in a group of young football players. This double-blind study included 22 male football players (mean = 19.96 ± 0.56), divided into two groups: supplemented and placebo. Before and after a 90-day period of training combined with supplementation (6 g of lyophilized black chokeberry extract), participants performed maximal multistage 20-m shuttle run tests at the beginning and at the end of the supplementation period, with blood sampled for analysis at different times before and after exercise. The levels of IL-6, IL-10, ferritin, myoglobin, hepcidin, 8-OHdG, albumin, and TAC were analyzed. The analysis of variance revealed a significant effect of 90-day supplementation with the lyophilized extract on changes in the IL-6 and IL-10 levels, and TAC induced by maximal aerobic effort. In conclusion, supplementation with lyophilized black chokeberry extract improves the performance and antioxidant status of serum in humans and induces protective changes in inflammatory markers.

Purpose: This study aimed to investigate the effects of 12 wk of omega-3 fatty acid supplementation during endurance training on omega-3 index (O3I) and indicators of running performance in amateur long-distance runners., Methods: Twenty-six amateur male long-distance runners ≥29 yr old supplemented omega-3 fatty acid capsules (OMEGA group, n = 14; 2234 mg of eicosapentaenoic acid and 916 mg of docosahexaenoic acid daily) or medium-chain triglycerides capsules as placebo (medium-chain triglyceride [MCT] group, n = 12; 4000 mg of MCT daily) during 12 wk of endurance training. Before and after intervention, blood samples were collected for O3I assessment, and an incremental test to exhaustion and a 1500-m run trial were performed., Results: O3I was significantly increased in the OMEGA group (from 5.8% to 11.6%, P < 0.0001). A significant increase in V̇O 2peak was observed in the OMEGA group (from 53.6 ± 4.4 to 56.0 ± 3.7 mL·kg -1 ⋅min -1 , P = 0.0219) without such change in MCT group (from 54.7 ± 6.8 to 56.4 ± 5.9 mL·kg -1 ⋅min -1 , P = 0.1308). A positive correlation between the change in O3I and the change in running economy was observed when data of participants from both groups were combined (-0.1808 ± 1.917, P = 0.0020), without such an effect in OMEGA group alone ( P = 0.1741). No effect of omega-3 supplementation on 1500-m run results was observed., Conclusions: Twelve weeks of omega-3 fatty acid supplementation at a dose of 2234 mg of eicosapentaenoic acid and 916 mg of docosahexaenoic acid daily during endurance training resulted in the improvement of O3I and running economy and increased V̇O 2peak without improvement in the 1500-m run trial time in amateur runners., (Copyright © 2022 The Author(s). Published by Wolters Kluwer Health, Inc. on behalf of the American College of Sports Medicine.)

It is not fully understood how supplementation with omega-3 fatty acids affects the metabolism of amino acids required for the bioavailability/synthesis of NO, i.e., L-arginine (L-arg), asymmetric dimethylarginine (ADMA), their metabolites, and the L-arg/ADMA ratio and their impact on running economy (RE) in runners. Thus, 26 male amateur endurance runners completed a twelve-week study in which they were divided into two supplemented groups: the OMEGA group (n = 14; 2234 mg and 916 mg of eicosapentaenoic and docosahexaenoic acid daily) or the MCT group (n = 12; 4000 mg of medium-chain triglycerides daily). At the same time, all participants followed an endurance training program. Before and after the 12-week intervention, blood was collected from participants at two time points (at rest and immediately post-exercise) to determine EPA and DHA in red blood cells (RBCs) and plasma levels of L-arg, ADMA, and their metabolites. RBC EPA and DHA significantly increased in the OMEGA group (p < 0.001), which was related to the resting increase in L-arg (p = 0.001) and in the L-arg/ADMA ratio (p = 0.005) with no changes in the MCT group. No differences were found in post-exercise amino acid levels. A total of 12 weeks of omega-3 fatty acid supplementation at a dose of 2234 mg of EPA and 916 mg of DHA daily increased levels of L-arg and the L-arg/ADMA ratio, which indirectly indicates increased bioavailability/NO synthesis. However, these changes were not associated with improved RE in male amateur endurance runners.

In humans, most free tryptophan is degraded via kynurenine pathways into kynurenines. Kynurenines modulate the immune system, central nervous system, and skeletal muscle bioenergetics. Consequently, kynurenine pathway metabolites (KPMs) have been studied in the context of exercise. However, the effect of vitamin D supplementation on exercise-induced changes in KPMs has not been investigated. Here, we analyzed the effect of a single high-dose vitamin D supplementation on KPMs and tryptophan levels in runners after an ultramarathon. In the study, 35 amateur runners were assigned into two groups: vitamin D supplementation group, administered 150,000 IU vitamin D in vegetable oil 24 h before the run ( n = 16); and control (placebo) group ( n = 19). Blood was collected for analysis 24 h before, immediately after, and 24 h after the run. Kynurenic, xanthurenic, quinolinic, and picolinic acids levels were significantly increased after the run in the control group, but the effect was blunted by vitamin D supplementation. Conversely, the decrease in serum tryptophan, tyrosine, and phenylalanine levels immediately after the run was more pronounced in the supplemented group than in the control. The 3-hydroxy-l-kynurenine levels were significantly increased in both groups after the run. We conclude that vitamin D supplementation affects ultramarathon-induced changes in tryptophan metabolism.

Vitamin C supplementation and exercise influence pro/antioxidative status and the cellular stress response. We tested the effects of exercise training for 6 weeks, supported by 1000 mg of vitamin C supplementation in elderly women. Thirty-six women were divided into two groups: a control group (CON) (n = 18, age 69.4 ± 6.4 years, 70.4 ±10.4 kg body mass) and a supplemented group (SUPP) (n = 18, aged 67.7 ± 5.6 years, body mass 71.46 ± 5.39 kg). Blood samples were taken twice (at baseline and 24 h after the whole period of training), in order to determine vitamin C concentration, the total oxidative status/capacity (TOS/TOC), total antioxidant status/capacity (TAS/TAC), and gene expression associated with cellular stress response: encoding heat shock factor ( HSF1 ), heat shock protein 70 ( HSPA1A ), heat shock protein 27 ( HSPB1 ), and tumor necrosis factor alpha ( TNF-α ). We observed a significant increase in TOS/TOC, TAS/TAC, and prooxidant/antioxidant balance in the SUPP group. There was a significant decrease in HSPA1A in the CON group and a different tendency in the expression of HSF1 and TNF-α between groups. In conclusion, vitamin C supplementation enhanced the pro-oxidation in elderly women with a normal plasma vitamin C concentration and influenced minor changes in training adaptation gene expression.

In this study, we aim to verify whether swim training can improve lactate metabolism, NAD+ and NADH levels, as well as modify the activity of glycolytic and NADH shuttle enzymes and monocarboxylate transporters (MCTs) in skeletal muscle of amyotrophic lateral sclerosis (ALS) mice. ALS mice (SOD1G93A) (n = 7 per group) were analyzed before the onset of ALS, at first disease symptoms (trained and untrained), and the last stage of disease (trained and untrained), and then compared with a wild-type (WT) group of mice. The blood lactate and the skeletal muscle concentration of lactate, NAD+ and NADH, MCT1 and MCT4 protein levels, as well as lactate dehydrogenase (LDH) and malate dehydrogenase (MDH) activities in skeletal muscle were determined by fluorometric, Western blotting, liquid chromatography-MS3 spectrometry, and spectrometric methods. In the untrained terminal ALS group, there were decreased blood lactate levels (p < 0.001) and increased skeletal muscle lactate levels (p < 0.05) as compared with a WT group of mice. The amount of nicotinamide adenine dinucleotides in the ALS groups were also significantly reduced as well as LDH activity and the level of MCT1. Swim training increased lactate levels in the blood (p < 0.05 vs. ALS TERMINAL untrained). In addition, cytosolic MDH activity and the cMDH/LDH 2.1 ratio were significantly higher in trained vs. untrained mice (p < 0.05). The data indicate significant dysfunction of lactate metabolism in ALS mice, associated with a reduction in muscle anaerobic metabolism and NADH transporting enzymes, as well as swim-induced compensation of energy demands in the ALS mice.