Acute effects of nitrate and breakfast on working memory, cerebral blood flow, arterial stiffness, and psychological factors in adolescents: Study protocol for a randomised crossover trial.

BACKGROUND: Inorganic nitrate has been shown to acutely improve working memory in adults, potentially by altering cerebral and peripheral vasculature. However, this remains unknown in adolescents. Furthermore, breakfast is important for overall health and psychological well-being. Therefore, this study will investigate the acute effects of nitrate and breakfast on working memory performance, task-related cerebral blood flow (CBF), arterial stiffness, and psychological outcomes in Swedish adolescents. METHODS: This randomised crossover trial will recruit at least 43 adolescents (13-15 years old). There will be three experimental breakfast conditions: (1) none, (2) low-nitrate (normal breakfast), and (3) high-nitrate (concentrated beetroot juice with normal breakfast). Working memory (n-back tests), CBF (task-related changes in oxygenated and deoxygenated haemoglobin in the prefrontal cortex), and arterial stiffness (pulse wave velocity and augmentation index) will be measured twice, immediately after breakfast and 130 min later. Measures of psychological factors and salivary nitrate/nitrite will be assessed once before the conditions and at two-time points after the conditions. DISCUSSION: This study will provide insight into the acute effects of nitrate and breakfast on working memory in adolescents and to what extent any such effects can be explained by changes in CBF. This study will also shed light upon whether oral intake of nitrate may acutely improve arterial stiffness and psychological well-being, in adolescents. Consequently, results will indicate if nitrate intake from beetroot juice or if breakfast itself could acutely improve cognitive, vascular, and psychological health in adolescents, which can affect academic performance and have implications for policies regarding school meals. TRIAL REGISTRATION: The trial has been prospectively registered on 21/02/2022 at https://doi.org/10.1186/ISRCTN16596056. Trial number: ISRCTN16596056.

Effects of prolonged sitting and physical activity breaks on measures of arterial stiffness and cortisol in adolescents.

AIM: In adults, prolonged periods of sitting have been linked to acute negative effects on vascular structure and function. The aim of this study was to evaluate the acute effects of physical activity (PA) breaks during prolonged sitting on arterial stiffness, cortisol and psychological factors in adolescents. METHODS: Adolescents underwent different short (3-min) breaks starting every 20 min, during 80 min of sitting on three separate days. Breaks were (A) social seated breaks (SOC), (B) low-intensity simple resistance activity PA breaks (SRA) and (C) moderate-intensity step-up PA breaks (STEP). The arterial stiffness measures were augmentation index (AIx), AIx@75 and pulse wave velocity (PWV). Cortisol was measured from saliva. Psychological factors were self-reported. RESULTS: Eleven girls and six boys (average age 13.6 ± 0.7 years) participated, with average baseline heart rates of 72 ± 11 bpm, systolic/diastolic blood pressure 111 ± 7/64 ± 6 mmHg and cortisol 10.9 ± 5.8 nmoL/L. PWV, cortisol and psychological factors did not change after any of the conditions. AIx@75 increased significantly (4.9 ± 8.7-9.2 ± 13.2) after the STEP intervention compared with SOC and SRA (time × condition p < 0.05). CONCLUSION: Arterial stiffness increased after prolonged sitting with frequent, short step-up activity breaks. The results indicate potential important intensity-dependent effects of physical activity on vascular regulation in youth.

ABBaH teens: Activity Breaks for Brain Health in adolescents: study protocol for a randomized crossover trial.

BACKGROUND: Physical activity breaks are widely being implemented in school settings as a solution to increase academic performance and reduce sitting time. However, the underlying physiological mechanisms suggested to improve cognitive function from physical activity and the frequency, intensity, and duration of the breaks remain unknown. This study will investigate the effects of frequent, short physical activity breaks during prolonged sitting on task-related prefrontal cerebral blood flow, cognitive performance, and psychological factors. Additionally, the moderating and mediating effects of arterial stiffness on changes in cerebral blood flow will be tested. METHODS: This is a protocol for a randomized crossover study that will recruit 16 adolescents (13-14 years old). Participants will undergo three different conditions in a randomized order, on three separate days, involving sitting 80 min with a different type of break every 17 min for 3 min. The breaks will consist of (1) seated social breaks, (2) simple resistance activities, and (3) step-up activities. Before and after the 80-min conditions, prefrontal cerebral blood flow changes will be measured using functional near-infrared spectroscopy (primary outcome), while performing working memory tasks (1-, 2-, and 3-back tests). Arterial stiffness (augmentation index and pulse wave velocity) and psychological factors will also be assessed pre and post the 80-min interventions. DISCUSSION: Publication of this protocol will help to increase rigor in science. The results will inform regarding the underlying mechanisms driving the association between physical activity breaks and cognitive performance. This information can be used for designing effective and feasible interventions to be implemented in schools. TRIAL REGISTRATION: www.ClinicalTrials.gov , NCT04552626 . Retrospectively registered on September 21, 2020.

Frequent, Short Physical Activity Breaks Reduce Prefrontal Cortex Activation but Preserve Working Memory in Middle-Aged Adults: ABBaH Study.

Prolonged sitting is increasingly common and may possibly be unfavorable for cognitive function and mood. In this randomized crossover study, the effects of frequent, short physical activity breaks during prolonged sitting on cognitive task-related activation of the prefrontal cortex were investigated. The effects on working memory, psychological factors, and blood glucose were also examined, and whether arterial stiffness moderated prefrontal cortex activation. Thirteen subjects (mean age 50.5 years; eight men) underwent three 3-h sitting conditions, interrupted every 30-min by a different 3-min break on separate, randomized-ordered days: seated social interactions (SOCIAL), walking (WALK), or simple resistance activities (SRA). Arterial stiffness was assessed at baseline. Before and after each 3-h condition, psychological factors (stress, mood, sleepiness, and alertness) were assessed through questionnaires and functional near-infrared spectroscopy (fNIRS) was used to measure changes in prefrontal oxygenated hemoglobin (Oxy-Hb), indicative of cortical activation, while performing working memory tasks [1- (baseline), 2-, and 3-back]. Blood glucose levels were continuously measured throughout the conditions. Results revealed no significant changes in Oxy-Hb during the 2-back compared with the 1-back test in any condition, and no time-by-condition interactions. During the 3-back test, there was a significant decrease in Oxy-Hb compared with the 1-back after the WALK condition in the right prefrontal cortex, but there were no time-by-condition interactions, although 3-back reaction time improved only in the WALK condition. Mood and alertness improved after the WALK condition, which was significantly different from the SOCIAL condition. Arterial stiffness moderated the effects, such that changes in Oxy-Hb were significantly different between WALK and SOCIAL conditions only among those with low arterial stiffness. Blood glucose during the interventions did not differ between conditions. Thus, breaking up prolonged sitting with frequent, short physical activity breaks may reduce right prefrontal cortex activation, with improvements in some aspects of working memory, mood, and alertness. Clinical Trial Registration:www.ClinicalTrials.gov, identifier NCT04137211.

Higher Total Physical Activity is Associated with Lower Arterial Stiffness in Swedish, Young Adults: The Cross-Sectional Lifestyle, Biomarkers, and Atherosclerosis Study.

PURPOSE: Arterial stiffness describes the rigidity of the arterial walls and is associated with risk factors for cardiovascular disease (CVD). Arterial stiffness predicts future events and mortality, and the predictive value is stronger in younger versus older subjects. The aims of the present study were, firstly, to present data on physical activity (PA) and time spent sedentary, in the population of Swedish, young adults. Secondly, to explore the association between PA and arterial stiffness. MATERIAL AND METHODS: Self-reported healthy, non-smoking, Swedish, young adults, 18-25 years old, participated in the cross-sectional Lifestyle, Biomarkers and Atherosclerosis (LBA) study. The daily PA was objectively measured with an accelerometer for 1 week. Of the 834 participants, 658 individuals had valid registrations. The arterial stiffness measures, pulse wave velocity (PWV) and augmentation index (AIx) were measured with applanation tonometry. RESULTS: Women were on overall more physically active than men, they spent 214 min/day in light PA (LPA) compared to men who spent 202 min/day. Women took significantly more steps per day than men, 7796 vs 7336 steps/day, and spent less time sedentary, 523 min/day, compared to men who spent 547 min/day sedentary. In total, 76% of the individuals spent on average at least 30 minutes per day in the recommended moderate and vigorous PA (MVPA). Lower arterial stiffness was associated with more MVPA and total PA in the total population. CONCLUSION: We conclude that in this age group of young, self-reported healthy adults 18-25 years, it is important to highlight the health-enhancing possibilities of time spent in physical activity on the vascular function, measured as PWV and AIx. It is of high relevance in a public health perspective to expand preventive efforts beyond the high-risk groups and encourage young adults to be physically active.

The importance of cardiorespiratory fitness and sleep duration in early CVD prevention: BMI, resting heart rate and questions about sleep patterns are suggested in risk assessment of young adults, 18-25 years : The cross-sectional lifestyle, biomarkers and atherosclerosis (LBA) study.

BACKGROUND: Cardiorespiratory fitness (CRF) and sleep habits are lifestyle factors with potential to prevent cardiovascular disease (CVD). CVD is the leading cause of death worldwide. It is therefore important to establish a healthy lifestyle at a young age. In the Lifestyle, Biomarkers and Atherosclerosis (LBA) study we have examined 834 healthy non-smoking adults, aged 18-25 years. The general purpose of the LBA study was to study the effect of lifestyle on traditional biomarkers known to influence CVD risk. The aims of the present study were to evaluate sleep habits of young adult women and men participating in the LBA study, and to compare the importance of sleep and other lifestyle habits on clinically relevant biomarkers for CVD. An additional aim was to find easy and reliable non-invasive biomarkers to detect young adults with increased risk of developing CVD later in life. METHODS: The participants had previously been examined for lifestyle factors, biomarkers and CVD risk score. They filled in a validated computerized questionnaire about their general physical and mental health. The questionnaire included questions on sleep duration and experienced quality of sleep. RESULTS: In total 27% of the young adult participants reported difficulties falling asleep or experienced troubled sleep with frequent awakenings per night. The experienced troubled sleep was not related to a higher CVD risk score, but sleep quality and duration were correlated. Shorter sleep duration was significantly associated to higher body mass index (BMI), body fat (%), homeostasis model assessment of insulin resistance (HOMA-IR) and CVD risk. The modifiable lifestyle factor with the highest odds ratio (OR) for CVD risk was CRF. Sleep duration was the second most influential lifestyle factor, more important than moderate- and vigorous physical activity (MVPA) and food habits. Correlations between CRF and heart rate (HR), (P < 0.01) and HOMA-IR and BMI (P < 0.01) were observed, indicating that BMI and resting HR in combination with questions about sleep patterns are easy and reliable non-invasive biomarkers to detect young adults who need counselling on a healthy lifestyle. CONCLUSION: Decreased sleep duration in combination with decreased CRF, in young adults, is a serious health issue.

ABBaH: Activity Breaks for Brain Health. A Protocol for a Randomized Crossover Trial.

Introduction: Extended periods of sitting may have detrimental effects on brain health. However, the effects of breaking up prolonged sedentary periods with frequent, short physical activity bouts on mechanisms to improve brain health remain unclear. Therefore, this study aims to investigate the immediate effects of uninterrupted sitting and frequent, short bouts of physical activity on cerebral blood flow and cognitive function in the prefrontal cortex in middle-aged adults. Methods: This is a protocol article to describe a randomized crossover study. We will collect data from 13 healthy adults, aged between 40 and 60 years old, with a body mass index <35 kg/m2. Participants will be required to come into the laboratory on three occasions, sit for 3 h, and perform a different type of break for 3 min every 30 min at each visit in a random order, being either: (1) a social break; (2) brisk walk on a treadmill; or (3) simple resistance activities. Before and after each experimental condition, cerebral blood flow (primary outcome) will be measured using functional near-infrared spectroscopy (fNIRS), with short-separation channels, and working memory (1-, 2-, and 3-back on the computer) will be assessed. The following additional secondary outcomes will be collected: psychological factors (questionnaires); arterial stiffness; salivary cortisol levels; and blood glucose levels. Conclusion: The results from this randomized crossover study will determine the effects of uninterrupted sitting and frequent, short bouts of physical activity on cerebral blood flow and cognitive performance. Publication of this study protocol emphasizes the importance of registration and publication of protocols in the field of sedentary behavior research.

Body composition is a strong predictor of local carotid stiffness in Swedish, young adults - the cross sectional Lifestyle, biomarkers, and atherosclerosis study.

BACKGROUND: Obesity has nearly tripled worldwide during the last four decades, especially in young adults, and is of growing concern since it is a risk factor for cardiovascular diseases (CVD). We explored how different body composition measurements are associated with intima media thickness (cIMT) and local stiffness in the common carotid artery, in a subsample of healthy, young women and men, from the Swedish Lifestyle, Biomarkers, and Atherosclerosis (LBA) Study. METHODS: From the LBA study, a subsample of 220 randomly selected, self-reported healthy individuals, 18-25 years old, were collected for the automatized local stiffness measurements; arterial distensibility, Young's elastic modulus, and ß stiffness index. Blood pressure and mean arterial pressure (MAP) was measured using automatic blood pressure equipment. Body mass index (BMI) was calculated, waist circumference was measured, and percentage of body fat assessed using an impedance body composition analyzer. The carotid artery was scanned by ultrasound and analyzed using B-mode edge wall tracking. cIMT was measured and local stiffness measurements were calculated with carotid blood pressure, measured with applanation tonometry. RESULTS: No association was found between cIMT and body composition. Local carotid stiffness was associated with body composition, and women had less stiff arteries than men (p < 0.001). Of the local stiffness measurements, arterial distensibility had the strongest associations with body composition measurements in both women and men (p < 0.05). Multiple regression analyses showed that BMI in women and BMI and percentage of body fat in men had the highest impact on arterial distensibility (p < 0.01 in both women and men). CONCLUSIONS: Arterial distensibility was the local stiffness measurement with the strongest associations to different body composition measurements, in both women and men. In this age group, body composition measurements seem to be stronger predictors of common carotid arterial stiffness than MAP, and is a convenient way of detecting young adults who need cardiovascular risk follow-up and lifestyle counseling.

Insulin resistance (HOMA-IR) and body fat (%) are associated to low intake of fruit and vegetables in Swedish, young adults: the cross-sectional lifestyle, biomarkers and atherosclerosis study.

BACKGROUND: In the cross-sectional Lifestyle, Biomarkers, and Atherosclerosis study (LBA study) we have previously reported a high prevalence (15%) of homeostasis model assessment of insulin resistance (HOMA-IR) in Swedish, young adults. The aim of the present study was to report the dietary habits of subjects 18.0-25.9 years, and to associate dietary habits to body composition measures; body mass index (BMI), body fat (%), waist circumference and to HOMA-IR, a risk marker for diabetes. METHOD: The subjects (577 women and 257 men) filled in a validated computerized food frequency questionnaire. The questionnaire was based on recommendations from the Swedish national food administration. To associate the dietary habits to BMI, body fat (%), waist circumference and to HOMA-IR the subjects were divided in two groups. Subjects "eating as recommended" and subjects "eating less/more than recommended". RESULTS: Recommended intake of fish and seafood (P < 0.05), fruit and vegetables (P < 0.001), and sweets (P < 0.05) were associated to lower HOMA-IR values compared to subjects not eating as recommended. When split by sex no difference in HOMA-IR was detected with recommended intake of fish and seafood, but women eating fish and seafood as recommended had less body fat (%) (P < 0.05) compared to women not eating fish and seafood as recommended. Recommended intake of fruit and vegetables was associated to lower HOMA-IR in women (P < 0.01), and in women and men to less body fat (%) (P < 0.05) compared to subjects not eating the recommended 500 g of fruit and vegetables per day. Both women and men with higher consumption of sweets than recommended had higher HOMA-IR (P < 0.05), but no difference in the body composition measures BMI, body fat (%) or waist circumference compared to subjects eating sweets as recommended. CONCLUSION: The results highlight the importance of reducing a high intake of sweets and to increase the intake of fish, fruit and vegetables, in young adults, to reduce the risk of future diabetes.

Individual variations in platelet reactivity towards ADP, epinephrine, collagen and nitric oxide, and the association to arterial function in young, healthy adults.

INTRODUCTION: Platelet aggregation and secretion can be induced by a large number of endogenous activators, such as collagen, adenosine diphosphate (ADP) and epinephrine. Conversely, the blood vessel endothelium constitutively release platelet inhibitors including nitric oxide (NO) and prostacyclin. NO and prostacyclin are also well-known vasodilators and contribute to alterations in local blood flow and systemic blood pressure. MATERIALS AND METHODS: In this study we investigated individual variations in platelet reactivity and arterial functions including blood pressure and flow-mediated vasodilation (FMD) in 43 young, healthy individuals participating in the Lifestyle, Biomarkers and Atherosclerosis (LBA) study. Platelet aggregation and dense granule secretion were measured simultaneously by light transmission and luminescence. FMD was measured with ultrasound. RESULTS: The platelet function assay showed inter-individual differences in platelet reactivity. Specifically, a sub-group of individuals had platelets with an increased response to low concentrations of ADP and epinephrine, but not collagen. When the NO-donor S-nitroso-N-acetyl-DL-penicillamine (SNAP) was combined with high doses of these platelet activators, the results indicated for sub-groups of NO-sensitive and NO-insensitive platelets. The individuals with NO-sensitive platelets in response to SNAP in combination with collagen had a higher capacity of FMD of the arteria brachialis. CONCLUSIONS: Platelet reactivity towards ADP, epinephrine and NO differs between young, healthy individuals. Some individuals have a more effective response towards NO, both in the aspect of platelet inhibition ex vivo, as well as vasodilation in vivo.

Arterial stiffness is associated to cardiorespiratory fitness and body mass index in young Swedish adults: The Lifestyle, Biomarkers, and Atherosclerosis study.

Background Early changes in the large muscular arteries are already associated with risk factors as hypertension and obesity in adolescence and young adulthood. The present study examines the association between arterial stiffness measurements, pulse wave velocity and augmentation index and lifestyle-related factors, body composition and cardiorespiratory fitness, in young, healthy, Swedish adults. Design This study used a population-based cross-sectional sample. Methods The 834 participants in the study were self-reported healthy, non-smoking, age 18-25 years. Augmentation index and pulse wave velocity were measured with applanation tonometry. Cardiorespiratory fitness was measured by ergometer bike test to estimate maximal oxygen uptake. Body mass index (kg/m2) was calculated and categorised according to classification by the World Health Organisation. Results Young Swedish adults with obesity and low cardiorespiratory fitness have significantly higher pulse wave velocity and augmentation index than non-obese young adults with medium or high cardiorespiratory fitness. The observed U-shaped association between pulse wave velocity and body mass index categories in women indicates that it might be more beneficial to be normal weight than underweight when assessing the arterial stiffness with pulse wave velocity. The highest mean pulse wave velocity was found in overweight/obese individuals with low cardiorespiratory fitness. The lowest mean pulse wave velocity was found in normal weight individuals with high cardiorespiratory fitness. Cardiorespiratory fitness had a stronger effect than body mass index on arterial stiffness in multiple regression analyses. Conclusions The inverse association between cardiorespiratory fitness and arterial stiffness is observed already in young adults. The study result highlights the importance of high cardiorespiratory fitness, but also that underweight individuals may be a possible risk group that needs to be further studied.

Aerobic fitness is associated with low cardiovascular disease risk: the impact of lifestyle on early risk factors for atherosclerosis in young healthy Swedish individuals - the Lifestyle, Biomarker, and Atherosclerosis study.

BACKGROUND: The progression of cardiovascular disease (CVD) and atherosclerosis is slow and develops over decades. In the cross-sectional Swedish Lifestyle, Biomarker, and Atherosclerosis study, 834 young, self-reported healthy adults aged 18.0-25.9 years have been studied to identify early risk factors for atherosclerosis. PURPOSE: The aims of this study were to 1) assess selected cardiometabolic biomarkers, carotid intima-media thickness (cIMT) as a marker of subclinical atherosclerosis, and lifestyle-related indicators (food habits, handgrip strength, and oxygen uptake, VO2 max); 2) analyze the associations between cIMT and lifestyle factors; and 3) identify subjects at risk of CVD using a risk score and to compare the characteristics of subjects with and without risk of CVD. METHOD: Blood samples were taken in a fasting state, and food habits were reported through a questionnaire. cIMT was measured by ultrasound, and VO2 max was measured by ergometer bike test. The risk score was calculated according to Wildman. RESULT: cIMT (mean ± standard deviation) was 0.50±0.06 mm, and VO2 max values were 37.8±8.5 and 42.9±9.9 mL/kg/min, in women and men, respectively. No correlation was found between aerobic fitness expressed as VO2 max (mL/kg/min) and cIMT. Using Wildman's definition, 12% of the subjects were classified as being at risk of CVD, and 15% had homeostasis model assessment of insulin resistance. A total of 35% of women and 25% of men had lower high-density lipoprotein cholesterol than recommended. Food habits did not differ between those at risk and those not at risk. However, aerobic fitness measured as VO2 max (mL/kg/min) differed; 47% of the subjects at risk had low aerobic fitness compared to 23% of the nonrisk subjects (P<0.001). CONCLUSION: High aerobic fitness is associated with low CVD risk in Swedish young adults. The high prevalence of young adults observed with unfavorable levels of high-density lipoprotein cholesterol and homeostasis model assessment of insulin resistance raises concerns about future CVD risk.

Improved Muscle Mitochondrial Capacity Following Gastric Bypass Surgery in Obese Subjects.

BACKGROUND: Weight loss resulting from low-calorie diets is often less than expected. We hypothesized that energy restriction would influence proton leakage and improve mitochondrial efficiency, leading to reduced energy expenditure, partly explaining the difficulties in weight loss maintenance. METHODS: Eleven women with a median BMI of 38.5 kg/m(2) (q-range 37-40), and referred to gastric bypass surgery participated. Before surgery, and at 6 months of follow-up, muscle biopsies were collected from the vastus lateralis muscle. Mitochondria were isolated and analyzed for coupled (state 3) and uncoupled (state 4) respiration and mitochondrial capacity (P/O ratio). RESULTS: At follow-up, the participants had a median BMI of 29.6 kg/m(2) (28.3-32.0). State 3 increased from 20.6 (17.9-28.9) to 34.9 nmol O2/min/U citrate synthase (CS) (27.0-49.0), p = 0.01, while state 4 increased from 2.8 (1.8-4.2) to 4.2 nmol O2/min/U CS (3.1-6.1), although not statistically significant. The P/O ratio increased from 2.7 (2.5-2.8) to 3.2 (3.0-3.4), p = 0.02, indicating improved mitochondrial efficiency. CONCLUSIONS: Six months after gastric bypass surgery, the mitochondrial capacity for coupled, i.e., ATP-generating, respiration increased, and the P/O ratio improved. Uncoupled respiration was not enhanced to the same extent. This could partly explain the decreased basal metabolism and the reduced inclination for weight loss during energy restriction.

Ultraendurance exercise increases the production of reactive oxygen species in isolated mitochondria from human skeletal muscle.

Exercise-induced oxidative stress is important for the muscular adaptation to training but may also cause muscle damage. We hypothesized that prolonged exercise would increase mitochondrial production of reactive oxygen species (ROS) measured in vitro and that this correlates with oxidative damage. Eight male athletes (24-32 yr) performed ultraendurance exercise (kayaking/running/cycling) with an average work intensity of 55% V(O(2peak)) for 24 h. Muscle biopsies were taken from vastus lateralis before exercise, immediately after exercise, and after 28 h of recovery. The production of H(2)O(2) was measured fluorometrically in isolated mitochondria with the Amplex red and peroxidase system. Succinate-supported mitochondrial H(2)O(2) production was significantly increased after exercise (73% higher, P = 0.025) but restored to the initial level at recovery. Plasma level of free fatty acids (FFA) increased fourfold and exceeded 1.2 mmol/l during the last 6 h of exercise. Plasma FFA at the end of exercise was significantly correlated to mitochondrial ROS production (r = 0.74, P < 0.05). Mitochondrial content of 4-hydroxy-nonenal-adducts (a marker of oxidative damage) was increased only after recovery and was not correlated with mitochondrial ROS production. Total thiol group level and glutathione peroxidase activity were elevated after recovery. In conclusion, ultraendurance exercise increases ROS production in isolated mitochondria, but this is reversed after 28 h recovery. Mitochondrial ROS production was not correlated with oxidative damage of mitochondrial proteins, which was increased at recovery but not immediately after exercise.

Reduced respiratory capacity in muscle mitochondria of obese subjects.

BACKGROUND/AIMS: The extent of weight gain varies among individuals despite equal calorie overconsumption. Furthermore, weight gain is often less than expected from energy excess. This suggests differences in metabolic efficiency and basal metabolism. Since mitochondrial uncoupling accounts for a substantial portion of the basal metabolic rate, we compared skeletal muscle mitochondrial respiration in obese subjects to normal-weight reference groups with various degrees of physical activity. METHODS: Muscle biopsies were taken from the vastus lateralis muscle of 9 healthy obese subjects (BMI 40 ± 3). Mitochondria were isolated and analyzed for coupled (state 3) and uncoupled (state 4) respirations as well as mitochondrial efficiency (P/O ratio) using pyruvate as a substrate. Respiratory data were compared to reference groups A, normal-weight untrained (BMI 24 ± 0.7), and B, normal-weight trained (BMI 24 ± 0.6). RESULTS: Obese subjects had a decreased respiratory capacity per mitochondrial volume compared to the reference groups: this was evident in state 4 (65% and 35% of reference group A and B, respectively) and state 3 (53% and 29% of A and B, respectively) (p < 0.05). CONCLUSION: Obese subjects had a low capacity for fuel oxidation, which may play a role in the predisposition of obesity. However, whether lower mitochondrial capacity is a cause or a consequence of obesity requires further research.

Four weeks of speed endurance training reduces energy expenditure during exercise and maintains muscle oxidative capacity despite a reduction in training volume.

We studied the effect of an alteration from regular endurance to speed endurance training on muscle oxidative capacity, capillarization, as well as energy expenditure during submaximal exercise and its relationship to mitochondrial uncoupling protein 3 (UCP3) in humans. Seventeen endurance-trained runners were assigned to either a speed endurance training (SET; n = 9) or a control (Con; n = 8) group. For a 4-wk intervention (IT) period, SET replaced the ordinary training ( approximately 45 km/wk) with frequent high-intensity sessions each consisting of 8-12 30-s sprint runs separated by 3 min of rest (5.7 +/- 0.1 km/wk) with additional 9.9 +/- 0.3 km/wk at low running speed, whereas Con continued the endurance training. After the IT period, oxygen uptake was 6.6, 7.6, 5.7, and 6.4% lower (P < 0.05) at running speeds of 11, 13, 14.5, and 16 km/h, respectively, in SET, whereas remained the same in Con. No changes in blood lactate during submaximal running were observed. After the IT period, the protein expression of skeletal muscle UCP3 tended to be higher in SET (34 +/- 6 vs. 47 +/- 7 arbitrary units; P = 0.06). Activity of muscle citrate synthase and 3-hydroxyacyl-CoA dehydrogenase, as well as maximal oxygen uptake and 10-km performance time, remained unaltered in both groups. In SET, the capillary-to-fiber ratio was the same before and after the IT period. The present study showed that speed endurance training reduces energy expenditure during submaximal exercise, which is not mediated by lowered mitochondrial UCP3 expression. Furthermore, speed endurance training can maintain muscle oxidative capacity, capillarization, and endurance performance in already trained individuals despite significant reduction in the amount of training.

Clinical variability of the 22q11.2 duplication syndrome.

The 22q11.2 duplication syndrome is an extremely variable disorder with a phenotype ranging from normal to learning disability and congenital defects. Both patients with a de novo 22q11.2 duplication and patients in whom the duplication has been inherited from a phenotypically normal parent have been reported. In this study we present two familial cases with a 3Mb 22q11.2 duplication detected by array-CGH. We also review the findings in 36 reported cases with the aim of delineating the phenotype of the 22q11.2 duplication syndrome. In a majority of the reported cases where parents have been tested, the duplication seems to have been inherited from a normal parent with minor abnormalities. With this in mind we recommend that family members of patients with a 22q11.2 duplication to be tested for this genetic defect.

Mitochondrial respiration is decreased in skeletal muscle of patients with type 2 diabetes.

We tested the hypothesis of a lower respiratory capacity per mitochondrion in skeletal muscle of type 2 diabetic patients compared with obese subjects. Muscle biopsies obtained from 10 obese type 2 diabetic and 8 obese nondiabetic male subjects were used for assessment of 3-hydroxy-Acyl-CoA-dehydrogenase (HAD) and citrate synthase activity, uncoupling protein (UCP)3 content, oxidative stress measured as 4-hydroxy-2-nonenal (HNE), fiber type distribution, and respiration in isolated mitochondria. Respiration was normalized to citrate synthase activity (mitochondrial content) in isolated mitochondria. Maximal ADP-stimulated respiration (state 3) with pyruvate plus malate and respiration through the electron transport chain (ETC) were reduced in type 2 diabetic patients, and the proportion of type 2X fibers were higher in type 2 diabetic patients compared with obese subjects (all P < 0.05). There were no differences in respiration with palmitoyl-l-carnitine plus malate, citrate synthase activity, HAD activity, UCP3 content, or oxidative stress measured as HNE between the groups. In the whole group, state 3 respiration with pyruvate plus malate and respiration through ETC were negatively associated with A1C, and the proportion of type 2X fibers correlated with markers of insulin resistance (P < 0.05). In conclusion, we provide evidence for a functional impairment in mitochondrial respiration and increased amount of type 2X fibers in muscle of type 2 diabetic patients. These alterations may contribute to the development of type 2 diabetes in humans with obesity.

Reduced efficiency, but increased fat oxidation, in mitochondria from human skeletal muscle after 24-h ultraendurance exercise.

The hypothesis that ultraendurance exercise influences muscle mitochondrial function has been investigated. Athletes in ultraendurance performance performed running, kayaking, and cycling at 60% of their peak O(2) consumption for 24 h. Muscle biopsies were taken preexercise (Pre-Ex), postexercise (Post-Ex), and after 28 h of recovery (Rec). Respiration was analyzed in isolated mitochondria during state 3 (coupled to ATP synthesis) and state 4 (noncoupled respiration), with fatty acids alone [palmitoyl carnitine (PC)] or together with pyruvate (Pyr). Electron transport chain activity was measured with NADH in permeabilized mitochondria. State 3 respiration with PC increased Post-Ex by 39 and 41% (P < 0.05) when related to mitochondrial protein and to electron transport chain activity, respectively. State 3 respiration with Pyr was not changed (P > 0.05). State 4 respiration with PC increased Post-Ex but was lower than Pre-Ex at Rec (P < 0.05 vs. Pre-Ex). Mitochondrial efficiency [amount of added ADP divided by oxygen consumed during state 3 (P/O ratio)] decreased Post-Ex by 9 and 6% (P < 0.05) with PC and PC + Pyr, respectively. P/O ratio remained reduced at Rec. Muscle uncoupling protein 3, measured with Western blotting, was not changed Post-Ex but tended to decrease at Rec (P = 0.07 vs. Pre-Ex). In conclusion, extreme endurance exercise decreases mitochondrial efficiency. This will increase oxygen demand and may partly explain the observed elevation in whole body oxygen consumption during standardized exercise (+13%). The increased mitochondrial capacity for PC oxidation indicates plasticity in substrate oxidation at the mitochondrial level, which may be of advantage during prolonged exercise.

Effects of acute and chronic endurance exercise on mitochondrial uncoupling in human skeletal muscle.

Mitochondrial proteins such as uncoupling protein 3 (UCP3) and adenine nucleotide translocase (ANT) may mediate back-leakage of protons and serve as uncouplers of oxidative phosphorylation. We hypothesized that UCP3 and ANT increase after prolonged exercise and/or endurance training, resulting in increased uncoupled respiration (UCR). Subjects were investigated with muscle biopsies before and after acute exercise (75 min of cycling at 70% of .VO2peak) or 6 weeks endurance training. Mitochondria were isolated and respiration measured in the absence (UCR or state 4) and presence of ADP (coupled respiration or state 3). Protein expression of UCP3 and ANT was measured with Western blotting. After endurance training, .VO2peak, citrate synthase activity (CS), state 3 respiration and ANT increased by 24, 47, 40 and 95%, respectively (all P < 0.05), whereas UCP3 remained unchanged. When expressed per unit of CS (a marker of mitochondrial volume) UCP3 and UCR decreased by 54% and 18%(P < 0.05). CS increased by 43% after acute exercise and remained elevated after 3 h of recovery (P < 0.05), whereas the other muscle parameters remained unchanged. An intriguing finding was that acute exercise reversibly enhanced the capacity of mitochondria to accumulate Ca2+(P < 0.05) before opening of permeability transition pores. In conclusion, UCP3 protein and UCR decrease after endurance training when related to mitochondrial volume. These changes may prevent excessive basal thermogenesis. Acute exercise enhances mitochondrial resistance to Ca2+ overload but does not influence UCR or protein expression of UCP3 and ANT. The increased Ca2+ resistance may prevent mitochondrial degradation and the mechanism needs to be further explored.