Search

Stress was previously defined as a response to any demand for change. These demands are noticeable during sport events, not only in athletes but also in coaches. Therefore, this study aimed to determine the influence of slow-paced breathing (SPB) on acute stress to handball coaches during official matches. One professional handball coach, aged 37 (height, 180 cm; weight, 80 kg), took part in the study. His salivary cortisol (C) concentration and alpha-amylase (AA) activity were measured, and his heart rate (HR) was monitored during six official matches. In the first three matches the SPB training protocol was not followed. From match four to match six, the coach performed SPB training every day and directly before the match. The decrease observed in AA was statistically significant with a large effect size (1.80). The trend of change in HR is visible and similar for matches with and without SPB. However, for SPB matches, there is a lower starting point compared to matches without SPB (89.81 ± 6.26 and 96.62 ± 8.10, respectively). Moreover, values of AA on SPB matches were smaller before the match (93.92 ± 15.89) compared to the same time point in non-SPB matches (115.30 ± 26.00). For AA, there is a significant effect size in the half-time (2.00) and after the match (−2.14). SPB matches showed a lower increase in AA. SPB might be used as one of the possible tools that could help coaches in achieving a desirable mental state during the match.

Stress plays a significant role in competitions and in the training of sports participants, and coaches are no exception. To better cope with stressful situations, close monitoring of coaches’ stress levels before, during, and after training and competitions is recommended. According to studies, the use of cortisol (C) and alpha-amylase (AA) as biomarkers for monitoring acute stress is recommended. Therefore, the aim of our study was to compare HR, salivary C and AA, and STAI scores before, during, and after handball matches and training sessions. The study examined one professional handball coach, aged 37, in stress markers (salivary cortisol (C) and alpha-amylase (AA) concentrations), heart rate (HR), and the State-Trait Anxiety Inventory (STAI) scores in five matches/training sessions in the First Qatar Handball League. Statistical analysis included the calculation of descriptive statistic parameters, Mann–Whitney U test for differences between match–training time points, and the effect size analysis (Cohen’s d) to calculate the magnitude of differences between match–training time points. Presented markers (C and AA) had statistically stronger reactions before, during, and after the matches than the corresponding time points of the training sessions, similar to HR data and STAI scores. Results indicate that, before and during the matches, the analyzed markers of stress increased, which might lead to the conclusion that coaches are more anxious than frightened before and during matches. Thus, stress-coping strategies for handball coaches should be more focused on stress anticipation and anxiety control.

Diverticular disease remains one of the most common conditions in the western world. Up to 25% of patients with diverticular disease require hospitalization, 15-30% of those of which require surgical intervention. CT scoring systems have been proposed as means to drive assessment and stratify patients necessitating hospital intervention. To assess and correlate CT scoring systems with clinical and surgical outcomes.Retrospective cohort analysis at a single institution. Single institutional assessment with patients presenting to emergency with a CT diagnosed episode of acute diverticulitis. One hundred and eighty-nine patients were included in the study, 61% of which were male. Patient demographics, comorbidities, medications, biochemistry and inflammatory markers, type of complication following acute diverticulitis, operative/procedural intervention, hospital outcome and mortality were measured. CT scoring systems assessed included modified Hinchey, modified Neff, World Society of Emergency Surgery (WSES) and modified Siewert scoring systems.Majority of patients had left-sided diverticulitis (91%) with localized air (88%) and pericolic abscess (49%) the most common radiological findings. 28% of patients required radiological and/or surgical management with 12% requiring intensive care unit (ICU) admission. There was a general trend for surgical/radiological intervention as the scores increased in severity. The four scoring systems were found to be statistically significant predictors of any intervention and of ICU admission with minimal statistical differences across the different scoring systems.Radiological CT scores for complicated diverticulitis are at best, moderate predictors of clinical and surgical outcomes and may serve to guide management with minimal statistical differences across different scores.

BACKGROUND: This study evaluated endocrine responsiveness (ER) to physical stress (contact vs. non-contact nature of play) during team handball matches, according to the playing positions, thereby contextualizing the contact nature of the handball match. METHODS: The participants were ten male team handball players (24.1±3.17 years, 188.2±6.42 cm, 94.6±9.6 kg) divided into two groups: contact playing positions (CPP) and non-contact playing positions (NCPP). To evaluate the ER, the salivary cortisol (C), testosterone (T), and alpha-amylase (AA) concentrations were assessed before the game, during the halftime break, and after the match. Moreover, playing time (PT) and the number of contacts (NC) were counted post-match by video analysis. To determine possible differences between PT and the NC in the first and second halves of the match, a paired-sample t-test was used. The differences among ER-measures were calculated by the magnitude-based Cohen’s effect size. Possible associations between NC and ER were analyzed by comparing CPP and NCPP in C, T, and AA. RESULTS: The CPP group performed significantly more physical contacts, while there was no difference in playing time between the groups. A stronger C response was evidenced in players with a longer playing time. During the game, the C response was directly determined by physical contact, with CPP players showing a stronger C response than NCPP players. CONCLUSIONS: This study provided evidence of the importance of contact actions during matches and training sessions, as a parameter of calculating training loads and preparing strategies for recovery and injury prevention. Further studies examining larger samples are warranted.

We aimed to determine the exercise intensity eliciting the highest (FATmax) and the lowest (FATmin) fat oxidation rate in sedentary men with obesity and to examine if these intensities correlate with their individual aerobic (AeT) and anaerobic (AnT) thresholds, respectively. Nineteen obese males performed breath-by-breath analysis to assess maximal oxygen consumption (VO2max) and to calculate their fat oxidation rate. Pearson correlation coefficient (r), coefficient of determination (R2) and paired t-test were used to evaluate VO2 at AeT and at FATmax and VO2 at AnT and at FATmin, respectively. FATmax and AeT occurred at 42.80±2.68% of VO2max and 43.02±2.73% of VO2max, while FATmin and AnT occurred at 53.40±3.65% of VO2max and 53.38±3.65% of VO2max, respectively. A high correlations were found between intensities matching FATmax and AeT (r=0.86, Pmin and at AnT (r=0.99, P

Кетеринг-индустријата е една од најдинамичните и најбрзорастечките бизниси во светот. Во индустријата за храна и пијалаци, кетеринг-индустријата убедливо го зазема првото местo во светот по динамичноста, но и по профитабилноста што ја овозможува. Кетерингот, односно планираното служење храна и пијалаци, е широко распространето во многу бизниси и индустрии, како што се на пр. Образовните институции, владите, медицинските установи, авионските летови, објектите за забава и рекреација итн. Кетеринг- индустријата, како и сите останати е подрачје коешто секојдневно подлежи на промени и за една кетеринг-компанија да остане конкурентна на пазарот неопходно е таа да ги следи најновите кетеринг-трендови, како и да одговори на најразличните барања за кетеринг на луѓето од современото општество. Во теоретскиот дел на овој докторски труд се дефинира кетерингот како поим, а исто така се дава и приказ на современите кетеринг-трендови кои се моментално присутни во светски рамки. Во однос на методолошката рамка, посебен дел во докажувањето на релевантноста на темата има примената на емпириското истражување со цел да се анализира и интерпретира главната тема, односно да се продлабочат постојните научни знаења, како и да се излезе со нови научни сознанија за условите и перспективите за развој на кетеринг-дејноста во Република Северна Македонија преку проучување на моменталната состојба на развиеност на кетеринг-дејност во земјава, како и преку проучување на најновите светски практики од оваа област. Притоа, се користи анкетен прашалник со структуирани, јасни и недвосмислени прашања, чија цел е да ја утврди моменталната состојба на развој на кетеринг-дејноста во Република Северна Македонија, со цел да се види ставот на испитаниците, воедно и сопственици, менаџери на угостителски објекти/кетеринг-компании, организатори на кетерингнастани (Event Managers) и oстанат персонал во угостителски објект/кетерингкомпанија вклучена во реализација на кетеринг-настани. Од главниот заклучок на дисертацијата може да се согледа дека Република Северна Македонија има голем потенцијал за развој на нови кетеринг-трендови и стилови кои во голема мера би влијаеле на развојот на економијата на земјата, како и на развој на туризмот. За да се постигне тоа во иднина, потребна е пред сѐ голема соработка помеѓу самиот персонал во угостителските објекти/кетерингкомпаниите, соработка со туристичките субјекти и надлежните тела во земјата, како и соработка на сите овие чинители на локално, регионално и меѓународно ниво.

BACKGROUND: Over the last half-century, different stoichiometric equations for calculating the energy cost of exercise based upon the combustion of mixtures of carbohydrates, fats, and proteins have been proposed and modified. With the means of indirect calorimetry, while measuring oxygen uptake, carbon dioxide production, and urinary urea nitrogen excretion, the contribution of specific substrates to overall energy production can be estimated. However, even with their long history of application, no previous studies have evaluated whether the use of different stoichiometric equations provides similar or distinct maximal fat oxidation rate (MFO) responses and information regarding MFO location (FA Tmax) in male athletes. METHODS: Twenty healthy male athletes performed graded exercise testing (GXT) cycle ergometry using breath by breath gas analysis to assess fat oxidation and maximal oxygen uptake. Analysis of variance followed by within-equation effects, within- equation factors, and post hoc pairwise comparisons were used to examine within- equation differences. RESULTS: Compared stoichiometric equations demonstrated significant differences in the mean and maximal fat oxidation rates, varying up to nearly 7%. FA Tmax differences, however, were not noticed. CONCLUSIONS: Our findings suggest that for within- study designs, the equation used appears to be less important, but when inter-study comparisons are planned, caution is in order due to the presence of inter-equation differences.

Over the past two decades, scientists have attempted to evaluate whether the point of maximal fat oxidation (FATmax) and the aerobic threshold (AerT) are connected. The existence of such a relationship would allow a more tailored training approach for athletes while improving the efficacy of individualized exercise prescriptions when treating numerous health-related issues. However, studies have reported conflicting results, and this issue remains unresolved. This systematic review and meta-analysis aimed: (i) to examine the strength of the association between FATmax and AerT by using the effect size (ES) of correlation coefficient (r) and standardized mean difference (SMD); (ii) to identify potential moderators and their influence on ES variability. This study was registered with PROSPERO (CRD42021239351) and ClinicalTrials (NCT03789045). PubMed and Google Scholar were searched and fourteen articles, consisting of overall 35 ES for r and 26 ES for SMD were included. Obtained ESs were analyzed using a multilevel random-effects meta-analysis. Our results support the presence of a significant association between FATmax and AerT exercise intensities. In conclusion, due to the large ES variance caused by clinical and methodological differences among the studies, we recommend that future studies follow strict standardization of data collection and analysis of FATmax and AerT-related outcomes.

Background: chicken meat extract is a popular functional food in Asia. It is rich in the bioactive compounds carnosine and anserine, two histidine-containing dipeptides (HCD). Studies suggest that acute pre-exercise ingestion of chicken extracts has important applications towards exercise performance and fatigue control, but the evidence is equivocal. This study aimed to evaluate the ergogenic potential of the pre-exercise ingestion of a homemade chicken broth (CB) vs a placebo soup on a short-lasting, high-intensity cycling exercise. Methods: fourteen men participated in this double-blind, placebo-controlled, crossover intervention study. Subjects ingested either CB, thereby receiving 46.4 mg/kg body weight of HCD, or a placebo soup (similar in taste without HCD) 40 min before an 8 min cycling time trial (TT) was performed. Venous blood samples were collected at arrival (fasted), before exercise and at 5 min recovery. Plasma HCD were measured with UPLC-MS/MS and glutathione (in red blood cells) was measured through HPLC. Capillary blood samples were collected at different timepoints before and after exercise. Results: a significant improvement (p=0.033; 5.2%) of the 8 min TT mean power was observed after CB supplementation compared to placebo. Post-exercise plasma carnosine (pConclusions: oral CB supplementation improved the 8 min TT performance albeit it did not affect the acid-base balance or oxidative status parameters. Further research should unravel the potential role and mechanisms of HCD, present in CB, in this ergogenic approach.

In this study, we aimed to determine the exercise intensities eliciting the highest (FATmax) and the lowest (FATmin) fat oxidation rate in male cyclists and to compare these intensities with their individual aerobic (AeT) and anaerobic (AnT) thresholds, respectively. Twenty-two moderately trained male cyclists performed a 2-min stage graded exercise test until exhaustion using breath-by-breath gas analysis to determine maximal oxygen consumption (VO2max). The fat oxidation rate was calculated using a stoichiometric equation, with metabolic thresholds being determined by ventilatory gas analysis. In the present group of subjects, FATmax was found at a 21.34 ± 3.64 ml·kg−1·min−1 corresponding to 45.05 ± 7.68% VO2max. AeT occurred at an exercise intensity of 22.15 ± 4.84 ml·kg−1·min−1, matching 46.76 ± 10.24% VO2max. AnT and FATmin were located at intensities equivalent to 32.56 ± 5.52 ml·kg−1·min−1 and 32.30 ± 5.35 ml·kg−1·min−1 which corresponded to 68.74 ± 11.65 and 68.19 ± 11.29% VO2max, respectively. The correlation between FATmax and AeT was strong (r = 0.80, p < 0.05). No statistical difference was observed between FATmin and AnT (r = 0.99, p < 0.05). The strong relationship between observed indices can be used to provide a more tailored exercise approach.

This study evaluated dynamics of testosterone, cortisol and alpha-amylase during a handball match in high-level male players. Ten male professional players (24.1 ± 3.1 years, 188.2 ± 6.4 cm, 94.6 ± 9.6 kg) were tested on salivary- testosterone, - cortisol and -alpha-amylase levels before (prematch), at halftime, and immediately after the game. Analysis of variance for repeated measurements (ANOVA) with consecutive post-hoc analyses and effect-size differences were calculated to identify differences between measurements. The associations among biomarkers were determined by Pearson's product moment correlation (Pearson's r). The ANOVA indicated significant differences in testosterone (F: 14.31, p < 0.01 ; significant post-hoc differences between prematch and remaining two measurements), and alpha-amylase (F: 9.78, p < 0.01 ; significant post-hoc differences between all measurements). Significant correlations were evidenced between: (i) alpha-amylase- and testosterone-changes during 1st halftime (Pearson's r: 0.81, p < 0.01), and (ii) alpha- amylase- and cortisol- changes during 2nd halftime (Pearson's r: 0.76, p < 0.05). The results indicated specific dynamics of biomarkers during the handball match, with significant increase in alpha-amylase during the entire match and significant increase in the testosterone level during the first half of the match. Further studies are needed to evaluate the associations between real-game performance and changes in hormonal responses.

Background: There is a shortage of longitudinal surveys in highly trained adolescent table tennis athletes reporting typical changes in performance related, hematological, and biochemical parameters. Objectives: The purpose of the study was to: (1) examine yearly variations of performance related parameters, and (2) report the variability of selected hematological and biochemical markers utilized to ascertain health status, and training influences in a group of young Arab table tennis athletes. Methods: Data were collected from ten male adolescent players with a mean age of 14.2 ± 1.3 years and a mean table tennis training experience of 6 years. Physical and biochemical measures were carried out at three important time points throughout the training season: a baseline measure (PRE - September); a mid-season (transition) measure (MID - February); an end of season measure (END - May). Results: Performance related parameters tended to significantly improve over time (P = 0.05), with lowest values displayed during PRE-measure. Hematocrit (P = 0.05), serum testosterone (P = 0.01), mean cell volume (MCV) (P = 0.001) and reticulocytes (RET) (P = 0.017) significantly increased at the END training period compared to PRE. Moreover, 25(OH)D levels (P = 0.001) showed a drop during MID to then significantly increase at the END whilst sex hormone binding globulin (SHBG; P = 0.005) decreased at the END period compared to MID. Conclusions: In conclusion, based on our findings, changes in certain physical parameters and, hematological and biochemical markers, take place during the training period. Therefore, coaches, sports scientists and nutritionists should take into consideration these fluctuations and plan and alter their training programs accordingly.

Plasma volume and red cell mass are key health markers used to monitor numerous disease states, such as heart failure, kidney disease, or sepsis. Nevertheless, there is currently no practically applicable method to easily measure absolute plasma or red cell volumes in a clinical setting. Here, a novel marker for plasma volume and red cell mass was developed through analysis of the observed variability caused by plasma volume shifts in common biochemical measures, selected based on their propensity to present with low variations over time. Once a month for 6 months, serum and whole blood samples were collected from 33 active males. Concurrently, the CO-rebreathing method was applied to determine target levels of hemoglobin mass (HbM) and blood volumes. The variability of 18 common chemistry markers and 27 Full Blood Count variables was investigated and matched to the observed plasma volume variation. After the removal of between-subject variations using a Bayesian model, multivariate analysis identified two sets of 8 and 15 biomarkers explaining 68% and 69% of plasma volume variance, respectively. The final multiparametric model contains a weighting function to allow for isolated abnormalities in single biomarkers. This proof-of-concept investigation describes a novel approach to estimate absolute vascular volumes, with a simple blood test. Despite the physiological instability of critically ill patients, it is hypothesized the model, with its multiparametric approach and weighting function, maintains the capacity to describe vascular volumes. This model has potential to transform volume management in clinical settings. Am. J. Hematol. 92:62-67, 2017. © 2016 Wiley Periodicals, Inc.

Although the role of vitamin D in calcium and bone metabolism is well documented, there is little clarity regarding the implications of low vitamin D status for inflammation, endothelial function, and antioxidant status in adolescent athletes. A prospective cohort study was conducted, and 44 male adolescent athletes, training at a sports academy in the Middle East, were assigned to either the intervention group (VitDs), consisting of vitamin D deficient athletes [twenty-five hydroxyvitamin D (25(OH)D) 30 ng/ml ; n = 22]. Vitamin D status, inflammatory cytokines, endothelium-related variables, and antioxidant enzymes were measured twice during a nine-week training period. At the baseline, the athletes in the VitDs group had significantly lower concentrations of 25(OH)D, vascular endothelial growth factor (VEGF), and glutathione peroxidase (GPx), and higher levels of parathyroid hormone (PTH), interleukin-6 (IL-6), interleukin-1 receptor antagonist (IL-1ra), and nitrite (NO2) (p < 0.05), in comparison to the athletes in the sufficient group. After vitamin D supplementation for the VitDs group, the two cohorts differed considerably in vitamin D binding protein (VDBP) and PTH concentrations (p < 0.05). Our data suggest that the low levels of vitamin D possibly induced alterations in the investigated biochemical parameters of athletes in the VitDs group at the beginning of the monitoring period. Furthermore, while the vitamin D supplementation was effective in increasing 25(OH)D status, it may have concurrently positively influenced variables that are related to inflammation, endothelial function, and enzymatic antioxidants.

BACKGROUND: The aim of this study was to identify the exercise intensity that elicited the highest rate of fat utilization (FA Tₘₐₓ) and to assess its relationship with the aerobic threshold (AeT) in male athletes. We hypothesized existence of high correlation of these two parameters when a short-staged graded treadmill test with AeT identified through breath-by-breath gas exchange analysis was used. METHODS: Fifty-six trained male athletes (age 25.6±3.4 y, height 197.8±5.6 cm, body mass 98.5±6.6 kg) participated in the study. Pearson correlation coefficient (r) and effect size (R2) were used to evaluate the existence of connection between VO₂ at AeT and at FA Tₘₐₓ. Maximal oxygen consumption (VO₂max) and substrate oxidation were determined using breath-by-breath indirect calorimetry during a short-staged graded treadmill test to exhaustion. RESULTS: Mean VO₂ₘₐₓwas 52.12±9.02 mL·kg⁻¹·min⁻¹. FA Tₘₐₓ and AeT occurred at 47.47±10.59% of VO₂ₘₐₓ and 45.95±10.21% of VO₂ₘₐₓ,respectively. Fat utilization at FA Tₘₐₓ was 0.59±0.24 g·min-1. A high correlation was found between VO₂ at FA Tₘₐₓ and at AeT (r=0.88, P

Autologous blood transfusion (ABT) has been frequently abused in endurance sport and is prohibited since the mid-1980s by the International Olympic Committee. Apart from any significant performance-enhancing effects, the ABT may pose a serious health issue due to aging erythrocyte-derived “red cell storage lesions.” The current study investigated the effect of blood storage in citrate phosphate dextrose adenine (CPDA1) on the red blood cell (RBC) membrane proteome. One unit of blood was collected in CPDA1 blood bags from 6 healthy female volunteers. RBC membrane protein samples were prepared on days 0, 14, and 35 of storage. Proteins were digested in gel and peptides separated by nanoliquid chromatography coupled to tandem mass spectrometry resulting in the confident identification of 33 proteins that quantitatively change during storage. Comparative proteomics suggested storage-induced translocation of cytoplasmic proteins to the membrane while redox proteomics analysis identified 14 proteins prone to storage-induced oxidation. The affected proteins are implicated in the RBC energy metabolism and membrane vesiculation and could contribute to the adverse posttransfusion outcomes. Spectrin alpha chain, band 3 protein, glyceraldehyde-3-phosphate dehydrogenase, and ankyrin-1 were the main proteins affected by storage. Although potential biomarkers of stored RBCs were identified, the stability and lifetime of these markers posttransfusion remain unknown. In summary, the study demonstrated the importance of studying storage-induced alterations in the erythrocyte membrane proteome and the need to understand the clearance kinetics of transfused erythrocytes and identified protein markers.

Background The aim of this study is to determine whether multidisciplinary team (MDT) meetings alter the length of time to treatment (LOTT) for patients with colorectal cancer. Methods We conducted a retrospective audit of all patients with colorectal cancer from the Geelong Hospital (TGH) mandatory colorectal database from 1 January 2006 to 3 February 2011. To be included, patients had to have had elective surgical intervention for primary colorectal adenocarcinoma. A comparison of historical controls was conducted between patients discussed in MDT meetings and those managed prior to the introduction of MDT meetings (3 October 2006) to determine the LOTT in days from definitive diagnosis (colonoscopy) to definitive management (surgery, radiotherapy or chemotherapy). Results In total, the median LOTT for the historical control and MDT era patient populations were 19.5 and 20 days, respectively. Within the MDT era, we noticed significantly longer times to treatment for patients with rectal cancer who were seen in an MDT meeting prior to definitive management than patients who did not have an intervening MDT meeting (P < 0.001). With a difference of 7.5 days, the clinical significance of these findings remains contentious. However, it is worthwhile recognizing this trend in patients who are exhibiting symptoms due to near obstruction or significant bleeding. The LOTT for colon cancer patients remained unchanged. Conclusion The introduction of MDT meetings to TGH has prolonged the LOTT for patients with rectal cancer. These findings pave the way for further revision of the efficiency of MDT meeting at TGH.

Energy is the most important resource in state-of-the-art Internet of Things solutions. There are a lot of concepts and techniques dedicated to save energy, mainly focused to reduce transmission, since the energy used for preprocessing (encoding) is incomparable smaller than energy used for broadcasting. If applications do not require real-time measurements, data compression is one solution to energy saving problem. The goal of this paper was to develop new coding scheme for delta compression, that can be used for efficient data compression of temporally correlated data, such as temperature measurements coming from different smart devices. We proved that our coding scheme can achieve up to 85% energy saving. Compared to other coding techniques, our scheme has greater compression ratio and lower memory requirements.

Plasma volume and red cell mass are key health markers used to monitor numerous disease states, such as heart failure, kidney disease, or sepsis. Nevertheless, there is currently no practically applicable method to easily measure absolute plasma or red cell volumes in a clinical setting. Here, a novel marker for plasma volume and red cell mass was developed through analysis of the observed variability caused by plasma volume shifts in common biochemical measures, selected based on their propensity to present with low variations over time. Once a month for 6 months, serum and whole blood samples were collected from 33 active males. Concurrently, the CO-rebreathing method was applied to determine target levels of hemoglobin mass (HbM) and blood volumes. The variability of 18 common chemistry markers and 27 Full Blood Count variables was investigated and matched to the observed plasma volume variation. After the removal of between-subject variations using a Bayesian model, multivariate analysis identified two sets of 8 and 15 biomarkers explaining 68% and 69% of plasma volume variance, respectively. The final multiparametric model contains a weighting function to allow for isolated abnormalities in single biomarkers. This proof-of-concept investigation describes a novel approach to estimate absolute vascular volumes, with a simple blood test. Despite the physiological instability of critically ill patients, it is hypothesized the model, with its multiparametric approach and weighting function, maintains the capacity to describe vascular volumes. This model has potential to transform volume management in clinical settings. Am. J. Hematol. 92:62-67, 2017. © 2016 Wiley Periodicals, Inc.

Purpose:To examine the effects of 3 d of intermittent fasting (3d-IF: abstaining from eating/drinking from dawn to sunset) on physical performance and metabolic responses to repeated sprints (RSs).Methods:Twenty-one active males performed an RS test (2 sets: 5 × 5-s maximal sprints with 25 s of recovery between and 3 min of recovery between sets on an instrumented treadmill) in 2 conditions: counterbalanced fed/control session (CS) and fasting session (FS). Biomechanical and biochemical markers were assessed preexercise and postexercise.Results:Significant main effects of IF were observed for sprints: maximal speed (P = .016), mean speed (P = .015), maximal power (P = .035), mean power (P = .049), vertical stiffness (P = .032), and vertical center-of-mass displacement (P = .047). Sprint speed and vertical stiffness decreased during the 1st (P = .003 and P = .005) and 2nd sprints (P = .046 and P = .048) of set 2, respectively. Postexercise insulin decreased in CS (P = .023) but not in FS (P = .230). Free-fatty-acid levels were higher in FS than in CS at preexercise (P < .001) and at postexercise (P = .009). High-density lipoprotein cholesterol (HDL-C) was higher at postexercise in FS (1.32 ± 0.22 mmol/L) than in CS (1.26 ± 0.21 mmol/L, P = .039). The triglyceride (TG) concentration was decreased in FS (P < .05) compared with CS.Conclusions:3d-IF impaired speed and power through a decrease in vertical stiffness during the initial runs of the 2nd set of RS. The findings of the current study confirmed the benefits of 3d-IF: improved HDL-C and TG profiles while maintaining total cholesterol and low-density lipoprotein cholesterol levels. Moreover, improving muscle power might be a key factor to retain a higher vertical stiffness and to partly counteract the negative effects of intermittent fasting.

Background: This study examined substrate oxidation at high-intensity exercise and aimed to determine when fat oxidation ends (FATmin). We hypothesized the existence of a connection between the anaerobic threshold (AnT) and FATmin point. Methods: Breath-by-breath data obtained from indirect calorimetry during a graded treadmill test were used to measure substrate oxidation and maximal oxygen uptake (VO2max) on 47 males (30 athletes (ATL) and 17 non-athletes (NATL)). Pearson correlation coefficient (r) and effect size (R2) were used to test correlations between VO2 at AnT and at FATmin. Results: Maximal oxygen uptake (VO2max) was 56.17 ± 4.95 and 46.04 ± 3.25 ml kg−1 min−1 in ATL and NATL, respectively. In ATL, AnT was observed at 87.57 ± 1.30 % of VO2max and FATmin was observed at 87.60 ± 1.60 % of VO2max. In NATL, AnT and FATmin were at 84.64 ± 1.10 % of VO2max and 85.25 ± 1.10 % of VO2max, respectively. Our data show large correlations between VO2 at AnT and VO2 at FATmin for ATL (r =0.99, p < 0.01, 95 % CI 0.99 to 1.00) and NATL (r= 0.97, p < 0.01, 95 % CI 0.91 to 0.98). The effect size of correlations for ATL and NATL were 0.98 and 0.94, respectively. Conclusions: Our results show high correlation between AnT and FATmin in both ATL and NATL with equal substrate oxidation rates at AnT.

Mature red blood cells (RBCs) are the end-stage of a development process that starts in the bone marrow and continues to differentiate, through reticulocyte stage, entering into the circulation with a four-month lifespan. While stored, RBCs undergo different changes. The aim of this study was to evaluate changes occurring in RBC membranes during storage that could be used as possible markers to detect the misuse of blood transfusion in sports. Whole blood was collected from two volunteers in blood bags and stored for 42 days at 4°C. At different times (1, 7, 21, and 42 days of storage) whole blood was extracted under sterile conditions and submitted to RBC membrane ghost preparation and further analysis. Proteomic methods were applied using two strategies: protein oriented using 2-DE gels and peptide oriented using isobaric tags for relative and absolute quantitation (iTRAQ). In both approaches, the goal was to compare detectable changes in RBC membrane proteome before and after standard storage at different times. Some of the changes were confirmed with both methodologies employed, while with others only with one of them. Complementarities of the methods in this case showed to be an advantage. Changes were observed in two different protein complexes. In one of them, changes consisted of proteins decreasing, while increasing in the other during storage of RBCs. They are mostly located in cytoskeleton--spectrin β, band 4.2, ankyrin-1, tropomodulin-1, β adducin, band 4.9 (dematin), tropomyosin, while some changes were also observed in transmembrane proteins (glycophorin C, aquaporin-1, band 3).

Receptor binding techniques have been widely used in different biochemical applications, with isolated membranes being the most used receptor preparation in this type of assays. In this study, intact cells were compared with isolated membranes as receptor support for radioligand receptor binding assay. The growth hormone secretagogue receptor 1a (GHSR-1a) expressed in human embryonic kidney 293 (HEK293) cells was used as a model of G-protein-coupled receptors. Differences between using intact cells in suspension and using isolated membranes were evaluated for different aspects of the receptor binding assay: total binding variations while both receptor preparations remain on ice, modifications in incubation conditions, saturation, and competition using different agonists. Intact cells are more prone to variability. Although under optimized settings both preparations were equivalent, the K(d) value for intact cells was three times higher than that using isolated membranes. However, no significant differences were observed in competition assays obtaining practically identical K(i) values for all ligands tested. For the GHSR-1a, isolated membranes are the better choice if particular incubation conditions are required (less variability), whereas intact cells yield easy, fast, and physiological conditions for receptor binding assays.

The haematological module of the Athlete's Biological Passport (ABP) has significantly impacted the prevalence of blood manipulations in elite sports. However, the ABP relies on a number of concentration-based markers of erythropoiesis, such as haemoglobin concentration ([Hb]), which are influenced by shifts in plasma volume (PV). Fluctuations in PV contribute to the majority of biological variance associated with volumetric ABP markers. Our laboratory recently identified a panel of common chemistry markers (from a simple blood test) capable of describing ca 67% of PV variance, presenting an applicable method to account for volume shifts within anti-doping practices. Here, this novel PV marker was included into the ABP adaptive model. Over a six-month period (one test per month), 33 healthy, active males provided blood samples and performed the CO-rebreathing method to record PV (control). In the final month participants performed a single maximal exercise effort to promote a PV shift (mean PV decrease -17%, 95% CI -9.75 to -18.13%). Applying the ABP adaptive model, individualized reference limits for [Hb] and the OFF-score were created, with and without the PV correction. With the PV correction, an average of 66% of [Hb] within-subject variance is explained, narrowing the predicted reference limits, and reducing the number of atypical ABP findings post-exercise. Despite an increase in sensitivity there was no observed loss of specificity with the addition of the PV correction. The novel PV marker presented here has the potential to improve the ABP's rate of correct doping detection by removing the confounding effects of PV variance.

Human growth hormone (hGH) is a protein endogenously produced predominantly by the anterior pituitary gland. Native hGH and, especially, its recombinant analogue (rhGH), used to treat patients with hormone deficiency, are supposed to be abused by athletes searching its anabolic and lipolytic effects. Hence, hGH use has been prohibited for a long time by the sport authorities, but until recently, hGH abuse could not be detected. Two approaches have been followed when trying to develop methods for GH abuse detection. The direct method identifies an abnormal ratio between GH isoforms—a result of hGH exogenous administration. The time window to find a cheating athlete by this approach is limited by the excretion time of the hormone. The indirect approach measures serum biomarkers directly affected by GH intake (eg, markers of released liver growth factors and of bone and collagen turnover). In this approach, the retrospective power extends further. Alternative possibilities for cheating related to hGH could be the administration of recombinant growth factors themselves, the administration of hGH metabolic precursors such as ghrelin-like GH secretagogues, or the genetic manipulation of muscle growth– related genes (gene doping). In parallel with the new types of abuse, which will surely emerge in the near future, the research and development for the improvement of the analytical detection of GH itself will continue.

Eosinophil cationic protein (ECP)/ribonuclease 3 is a member of the RNase A superfamily involved in inflammatory processes mediated by eosinophils. ECP is bactericidal, helminthotoxic, and cytotoxic to tracheal epithelium cells and to several mammalian cell lines although its RNase activity is low. We studied the thermal stability of ECP by fourth-derivative UV absorbance spectra, circular dichroism, differential scanning calorimetry, and Fourier transform infrared spectroscopy. The T (1/2) values obtained with the different techniques were in very good agreement (T (1/2) approximately 72 degrees C), and the stability was maintained in the pH range between 5 and 7. The ECP calorimetric melting curve showed, in addition to the main transition, a pretransitional conformational change with a T (1/2) of 44 degrees C. Both calorimetric transitions disappeared after successive re-heatings, and the ratio DeltaH versus DeltaH (vH) of 2.2 indicated a significant deviation from the two-state model. It was observed that the thermal unfolding was irreversible. The unfolding process gives rise to changes in the environment of aromatic amino acids that are partially maintained in the refolded protein with the loss of secondary structure and the formation of oligomers. From the thermodynamic analysis of ECP variants, the contribution of specific amino acids, such as Trp10 and the region 115-122, to thermal stability was also determined. The high thermal stability of ECP may contribute to its resistance to degradation when the protein is secreted to the extracellular medium during the immune response.

Eosinophil cationic protein (ECP) is a component of the eosinophil granule matrix. It shows marked toxicity against helminth parasites, bacteria single-stranded RNA viruses, and host epithelial cells. Secretion of human ECP is related to eosinophil-associated allergic, asthmatic, and inflammatory diseases. ECP belongs to the pancreatic ribonuclease superfamily of proteins, and the crystal structure of ECP in the unliganded form (determined previously) exhibited a conserved RNase A fold [Boix, E., et al. (1999) Biochemistry 38, 16794-16801]. We have now determined a high-resolution (2.0 A) crystal structure of ECP in complex with adenosine 2',5'-diphosphate (2',5'-ADP) which has revealed the details of the ribonucleolytic active site. Residues Gln-14, His-15, and Lys-38 make hydrogen bond interactions with the phosphate at the P(1) site, while His-128 interacts with the purine ring at the B(2) site. A new phosphate binding site, P(-)(1), has been identified which involves Arg-34. This study is the first detailed structural analysis of the nucleotide recognition site in ECP and provides a starting point for the understanding of its substrate specificity and low catalytic efficiency compared with that of the eosinophil-derived neurotoxin (EDN), a close homologue.

More than 30 years ago people have learned that exercise promotes oxidative damage in human tissues. Since then, our knowledge on exercise-induced free radical production and their effect has advanced markedly. In the beginning research has mainly focused on detrimental effects of exercise-induced free radical production (e.g. oxidation of macromolecules); however today a new era of redox biology exist that centers on the cell-signaling effects of exercise-induced free radicals, such as reactive oxygen species (ROS) that were shown to play a crucial role in muscle adaptation to exercise. Erythrocytes are rich in membrane polyunsaturated free fatty acid content and have high cellular concentrations of oxygen and haemoglobin that makes them susceptible to ROS induced damage. However, mild oxidative stress has beneficial effect by increasing endogenous antioxidant mechanisms. Thus, in the present work we have used sensitive mass spectrometry techniques to assess the potentially beneficial oxidative modifications of erythrocyte membrane proteins in response to exercise. We have recruited 3 healthy volunteers and their blood was drawn before exercise, immediately after and few hours after exercise. Erythrocyte membranes ("ghosts") were prepared and membrane proteins isolated. Proteins were then separated by 1-D gel electrophoresis, digested with trypsin and peptides analyzed by Orbitrap ELITE coupled with the Easy n-LC II for nano LC-MS/MS. The acquired data was searched with Proteome Discoverer 1.4 against Homo sapiens database to reveal the changes in the erythrocyte membrane proteome. Furthermore, data acquired was also searched through the database using the comprehensive workflow that included several dynamic modifications of peptides, including N-terminal carboxymethyl, C-terminal oxidation and terminal independent oxidation and trioxidation of peptides. The results obtained revealed several proteins that have translocated from the cytoplasm into the membrane following exercise. Moreover, numerous proteins showed exercise dependent modifications and up/down regulation. Exercise induced proteome changes were demonstrated to play an important role in several signaling pathways related to healthy living and lifestyle.

Objectives The substrates utilisation at high intensity exercise has been less studied and intensity at which carbohydrates (CHO) remain single energy source while fat utilisation becomes negligible (FAT min ), is debatable 4 . Since physical activity in aerobic fat burn zone is prescribed to facilitate improvement of fat oxidation and insulin sensitivity while improving health 1 , activity equal or above FAT min can’t be considered as beneficial. Therefore, FAT min could become important exercise parameter in sedentary population. The aim of this study was to determine FAT min while performing incremental treadmill test till exhaustion and to assess its correlation with anaerobic threshold (AT), which in turn can be used as an individuals performance marker or last exercise beneficial point. Methodology Thirteen sedentary females (age: 34 ± 3 y, body mass: 62 ± 4 kg, height: 165 ± 4 cm) performed breath-by-breath test to measure maximal oxygen uptake (VO 2max; ml·kg -1 ·min -1 ) and substrates oxidation (g·min -1 ). A Pearson correlation coefficient ( r ), 95% confidence interval ( CI ) and effect size ( R 2 ) were used to test connexion between VO 2 at AT and at FAT min . Alpha intervals were set at p 4 . The AT was detected by V-slope method 2 . Results VO 2max was 34.89 ± 4.34 ml·kg -1 ·min -1 (95% CI 34.02 to 35.52). FAT min and AT occurred at 24.53 ± 4.25 ml·kg -1 ·min -1 VO 2 (95% CI 24.29 to 24.97) and 24.04 ± 4.07 ml·kg -1 ·min -1 VO 2 (95% CI 24.01 to 24.80), respectively. A high correlation was found between VO 2 at FAT min and at AT ( r = 0.91, p R 2 was 0.83. Absolute fat oxidation rate at FAT min was 0.01 ± 0.01 g·min -1 (95% CI 0.00 to 0.01). Conclusions This is the first study to detect FAT min and demonstrate correlation between VO 2 at FAT min and at AT in sedentary females. High inter-subjects FAT min variation demonstrates marked degree of individuality and importance of existing correlation should be taken into consideration when aiming to optimise exercise prescription for sedentary population. References American College of Sports Medicine: Position Stand. “The Recommended Quantity and Quality of Exercise for Developing and Maintaining Cardiorespiratory and Muscular Fitness”. Med Sci Sports Exerc 1998; 30 :975–991. Beaver WL, Wasserman K, Whipp BJ. A new method for detecting anaerobic threshold by gas exchange. J Appl Physiol 1986; 60 (6):2020–2027. Elia M, Livesey G. Energy expenditure and fuel selection in biological systems: the theory and practice of calculations based on indirect calorimetry and tracer methods. World Rev Nutr Diet 1992; 70 :68–131. Peric R, Meucci M, Nikolovski Z. Fat utilisationutilization during high intensity exercise: when does it end? Sport Med Open 2016; 2 (35). DOI: 10.1186/s40798-016-0060-1

Background & Objectives: Both exercise and hypoxia acutely elicit changes in systemic growth factors and cytokines that mediate increases in erythropoiesis, angiogenesis and, perhaps, arteriogenesis. To what extent the down-stream responses to these interventions are inter-related is, as yet, unclear. A role for circulating micro RNAs (cmiRNAs) in the regulation of these processes is a novel prospect. Thus the aims of this study are to investigate the effect of hypoxia and exercise training on relevant growth factors and cmiRNAs in a healthy human cohort. Methods: 18 healthy, Caucasian, male, elite athletes participated in this study. The project was designed as a parallel, two-groups, matched, randomized, longitudinal (pre-, mid-, post-test), two week intervention. One group was exposed to hypoxia (15 hours/day) the other stayed in a normoxic environment. Both groups performed physical exercise in a 10°C hotter environment than usual. The exercise was standardized for both groups and the daily work load was monitored. Blood was sampled under fasting conditions in the morning, to avoid any effects of acute exercise. Hemoglobin (Hb), reticulocytes, erythropoietin, vascular endothelial growth factor (VEGF), interleukin 6 (IL-6), cmiRNAs 144, 146a, 221, 222 were tested in blood and sera. Results: In the normoxic group decreases from pre- to mid- and post-test were found for Hb (p=0.011, p=0.002) and IL-6 (p=0.028, p=0.008). However, in the hypoxic group several changes were apparent. In this group Hb changed at the pre- to mid-test (p=0.013), reticulocytes also increased from pre- to mid-test (p=0.066), erythropoietin increased from pre- to mid- and post-test (p=0.008, p=0.008). VEGF also showed a tendency to increase from pre- to mid-test (p=0.085) and increased from pre- to post-test (p=0.011). The 2-ΔΔCT values for cmiRNAs 221 and 222 showed an increasing trend at the post-test time point but did not reach statistical significance. Conclusions: Hypoxia stimulated several cytokines and growth factors mediating erythropoiesis and angiogenesis, which might have counteracted the plasma volume related decrease in Hb in the normoxic group. While there were changes in cmiRNAs (221 and 222) these were not statistically significant, perhaps due the small study cohort, which is currently being expanded to confirm these results.

Mature red blood cells (RBCs) are the end-stage of a development process that starts in the bone marrow and continues to differentiate, through reticulocyte stage, entering into the circulation with a four-month lifespan. While stored, RBCs undergo different changes. The aim of this study was to evaluate changes occurring in RBC membranes during storage that could be used as possible markers to detect the misuse of blood transfusion in sports. Whole blood was collected from two volunteers in blood bags and stored for 42 days at 4°C. At different times (1, 7, 21, and 42 days of storage) whole blood was extracted under sterile conditions and submitted to RBC membrane ghost preparation and further analysis. Proteomic methods were applied using two strategies: protein oriented using 2-DE gels and peptide oriented using isobaric tags for relative and absolute quantitation (iTRAQ). In both approaches, the goal was to compare detectable changes in RBC membrane proteome before and after standard storage at different times. Some of the changes were confirmed with both methodologies employed, while with others only with one of them. Complementarities of the methods in this case showed to be an advantage. Changes were observed in two different protein complexes. In one of them, changes consisted of proteins decreasing, while increasing in the other during storage of RBCs. They are mostly located in cytoskeleton--spectrin β, band 4.2, ankyrin-1, tropomodulin-1, β adducin, band 4.9 (dematin), tropomyosin, while some changes were also observed in transmembrane proteins (glycophorin C, aquaporin-1, band 3).

Growth hormone secretagogues (GHS) constitute a new GH deficiency treatment increasing exponentially in number and improved potency and bioavailability over the last decade. The growth hormone releasing activity makes these compounds attractive for the artificial improvement of the human sports skills, now that recombinant human growth hormone (rhGH) administration is effectively detected. The GHS family is extremely diverse both in number and chemical heterogeneity and keeps growing continuously. In this paper, a general screening test is proposed. To develop a universal method, the single common property of growth hormone secretagogues has been targeted: their capacity to bind to the GHS receptor 1a (GHS-R1a). Pretreated urine samples have been tested in a competition assay where eventually the GHS presence detached a radiolabelled ligand from the receptor in a dose-dependent manner. Blank urine samples were processed to determine potential age, gender and exercise effects, and to define a threshold beyond which a specimen is considered positive. Samples from a growth hormone releasing peptide 2 (GHRP-2) excretion study corroborated the screening assay applicability with a detection window of approximately 4.5 h, and results were confirmed by comparison with a dedicated LC-MS quantification of the intact compound.

With the use of a high yield prokaryotic expression system, large amounts of human eosinophil cationic protein (ECP) have been obtained. This has allowed a thorough kinetic study of the ribonuclease activity of this protein. The catalytic efficiencies for oligouridylic acids of the type (Up)nU>p, mononucleotides U>p and C>p, and dinucleoside monophosphates CpA, UpA, and UpG have been interpreted by the specific subsites distribution in ECP. The distribution of products derived from digestion of high molecular mass substrates, such as poly(U) and poly(C), by ECP was compared with that of RNase A. The characteristic cleavage pattern of polynucleotides by ECP suggests that an exonucleaselike mechanism is predominantly favored in comparison to the endonuclease catalytic mechanism of RNase A. Comparative molecular modeling with bovine pancreatic RNase A-substrate analog crystal complexes revealed important differences in the subsite structure, whereas the secondary phosphate-binding site (p2) is lacking, the secondary base subsite (B2) is severely impaired, and there are new interactions at the po, Bo, and p-1 sites, located upstream of the P-O-5* cleavable phosphodiester bond, that are not found in RNase A. The differences in the multisubsites structure could explain the reduced catalytic efficiency of ECP and the shift from an endonuclease to an exonuclease-type mechanism.

Eosinophil cationic protein (ECP) is located in the matrix of the eosinophil's large specific granule and has marked toxicity for a variety of helminth parasites, hemoflagellates, bacteria, single-stranded RNA virus, and mammalian cells and tissues. It belongs to the bovine pancreatic ribonuclease A (RNase A) family and exhibits ribonucleolytic activity which is about 100-fold lower than that of a related eosinophil ribonuclease, the eosinophil-derived neurotoxin (EDN). The crystal structure of human ECP, determined at 2.4 A, is similar to that of RNase A and EDN. It reveals that residues Gln-14, His-15, Lys-38, Thr-42, and His-128 at the active site are conserved as in all other RNase A homologues. Nevertheless, evidence for considerable divergence of ECP is also implicit in the structure. Amino acid residues Arg-7, Trp-10, Asn-39, His-64, and His-82 appear to play a key part in the substrate specificity and low catalytic activity of ECP. The structure also shows how the cationic residues are distributed on the surface of the ECP molecule that may have implications for an understanding of the cytotoxicity of this enzyme.

Gene therapy is anticipated as being an important medical development. Essential to its effectiveness is the appropriate activity (protein expression) in the expected target cells. A noninvasive diagnostic procedure of successful gene expression will be of paramount importance to validate its use or its misuse (eg, sports gene doping). Externally detectable labeled oligonucleotide hybridizing with the messenger RNA generated by the transferred gene has been proposed as a possibility to monitor successful gene therapy. The authors selected the erythropoietin gene (Epo) for a pilot study on erythropoietin protein expression in mouse muscle. Oligonucleotides of peptide nucleic acid (PNA) type capable of antisense binding to unique murine Epo-mRNA sequences were synthesized by solid phase methods, and elongated at the N-terminus with the HIV Tat (48–60) cell penetrating peptide. They were labeled with fluorescence and radioactive tags to verify penetration and longer half-life properties in Epo gene transfected C2C12 mouse muscle cells as compared with corresponding wild- type cells. Downregulation of newly expressed erythropoietin protein in such cells additionally confirmed the penetration and hybridizing properties of the selected labeled oligonucleotide. 123I-labeled Tat-PNAs were intravenously injected into mice that had previously received the Epo gene into the right tibialis muscle by DNA electrotransfer. Preferential accumulation of radioactivity in the transferred limb as compared with the contralateral limb was ascertained, especially for 123I-Tat-CTA CGT AGA CCA CT (labeled Tat-PNA 1). This study provides experimental data to support the potential use of external noninvasive image detection to monitor gene therapy. The extension of the approach to more sensitive methods for whole-body external detection such as positron emission tomography appears feasible.

The eosinophil cationic protein (ECP) is a basic secretion proteininvolved in the immune response system. ECP levels in biological fluidsare an indicator of eosinophil-specific activation and degranulationand are currently used for the clinical monitoring and diagnosis ofinflammatory disorders. A polyclonal epitope-specific antibody has beenobtained by immunizing rabbits with a conjugated synthetic peptide. Asequence corresponding to a large exposed loop in the human ECPthree-dimensional structure (D115–Y122) was selected as a putativeantigenic epitope. The antibody was purified on an affinity columnusing recombinant ECP (rECP) as antigen. The antibody (D112–P123 Ab)specifically recognizes rECP and its native glycosylated andnonglycosylated forms in plasma, granulocytes, and sputum. The antibodydetects as little as 1 ng of rECP, can be used both in reducing andnonreducing conditions, and does not cross-react with the highlyhomologous eosinophil-derived neurotoxin or other proteins of thepancreatic ribonuclease superfamily.