Trio RNA sequencing in a cohort of medically complex children.

Genome sequencing (GS) is a powerful test for the diagnosis of rare genetic disorders. Although GS can enumerate most non-coding variation, determining which non-coding variants are disease-causing is challenging. RNA sequencing (RNA-seq) has emerged as an important tool to help address this issue, but its diagnostic utility remains understudied, and the added value of a trio design is unknown. We performed GS plus RNA-seq from blood using an automated clinical-grade high-throughput platform on 97 individuals from 39 families where the proband was a child with unexplained medical complexity. RNA-seq was an effective adjunct test when paired with GS. It enabled clarification of putative splice variants in three families, but it did not reveal variants not already identified by GS analysis. Trio RNA-seq decreased the number of candidates requiring manual review when filtering for de novo dominant disease-causing variants, allowing for the exclusion of 16% of gene-expression outliers and 27% of allele-specific-expression outliers. However, clear diagnostic benefit from the trio design was not observed. Blood-based RNA-seq can facilitate genome analysis in children with suspected undiagnosed genetic disease. In contrast to DNA sequencing, the clinical advantages of a trio RNA-seq design may be more limited.

Correction: Klentrou et al. Circulating Levels of Bone Markers after Short-Term Intense Training with Increased Dairy Consumption in Adolescent Female Athletes. Children 2021, 8, 961.

There was an error in the original publication [...].

Bone Turnover Markers and Osteokines in Adolescent Female Athletes of High- and Low-Impact Sports Compared With Nonathletic Controls.

This study examined differences in resting concentrations of markers of bone formation and resorption, and osteokines between female adolescent (12-16 y) swimmers, soccer players, and nonathletic controls. Resting, morning blood samples were obtained after an overnight fast from 20 swimmers, 20 soccer players, and 20 nonathletic controls, matched for age. carboxyl-terminal cross-linking telopeptide of type I collagen (CTX), amino-terminal propeptide of type I collagen (P1NP), total osteocalcin (OC), sclerostin, osteoprotegerin (OPG), and receptor activator of nuclear factor kappa B ligand (RANKL) were analyzed in serum. After controlling for percent body fat, there were no significant differences between swimmers and nonathletic controls in any of the measured markers. In contrast, soccer players had significantly higher P1NP (89.5 [25.6] ng·mL-1), OC (57.6 [22.9] ng·mL-1), and OPG (1052.5 [612.6] pg·mL-1) compared with both swimmers (P1NP: 66.5 [20.9] ng·mL-1; OC: 24.9 [12.5] ng·mL-1; OPG: 275.2 [83.8] pg·mL-1) and controls (P1NP: 58.5 [16.2] ng·mL-1; OC: 23.2 [11.9] ng·mL-1; OPG: 265.4 [97.6] pg·mL-1), with no differences in CTX, sclerostin, and RANKL. These results suggest that bone formation is higher in adolescent females engaged in high-impact sports like soccer compared with swimmers and controls.

Correction: McKinlay et al. Effects of Post-Exercise Whey Protein Consumption on Recovery Indices in Adolescent Swimmers. Int. J. Environ. Res. Public Health 2020, 17, 7761.

The authors of "Effects of Post-Exercise Whey Protein Consumption on Recovery Indices in Adolescent Swimmers" report an error in Table 1 of their article [...].

Intensified training in adolescent female athletes: a crossover study of Greek yogurt effects on indices of recovery.

Background: During a period of intensified exercise (e.g. training/identification camps), often undertaken by competitive youth athletes, the maintenance of muscle function and peak performance can become challenging due to an accumulation of fatigue. The provision of post-exercise dairy protein in adults has been previously shown to accelerate recovery; however, its efficacy in youth athletes is currently unknown. Therefore, the purpose of this study was to examine the effects of increased dairy protein consumption with plain Greek yogurt (GY) on performance and recovery indices during an intensified soccer training camp in adolescent female soccer players. Methods: Thirteen players (14.3 ± 1.3 years) participated in a randomized, double blinded, crossover design study where they received 3 servings/day of either GY (~115 kcal, 17 g protein, ~11.5 g carbohydrates) or an isoenergetic carbohydrate control (CHO, ~115 kcal, 0.04 g protein, ~28.6 g carbohydrates) during two 5-day soccer-specific training camps. Performance was assessed before and after each training camp. Fasted, morning, creatine kinase (CK), insulin-like growth factor-1 (IGF-1), C-reactive protein (CRP), interleukin 6 (IL6), interleukin 10 (IL10) and tumor necrosis factor-α (TNFα) were measured in plasma pre- and post-training. Results: Training led to decrements in counter-movement jump (p = 0.01), broad jump (p = 0.04) and aerobic capacity (p = 0.006), with no effect of GY. A significant increase in anti-inflammatory cytokine IL10 was observed from pre- to post-training in GY (+26% [p = 0.008]) but not in CHO (p = 0.89). CRP and CK increased (+65% [p = 0.005] and +119% [p ≤ 0.001], respectively), while IGF-1 decreased (-34% [p ≤ 0.001]) from pre- to post-training with no difference between conditions. Conclusions: These results demonstrate that consumption of GY did not offer any added recovery benefit with respect to measures of performance and in the attenuation of exercise-induced muscle damage above that achieved with energy-matched carbohydrate in this group of young female soccer players. However, regular consumption of GY may assist with the acute anti-inflammatory response during periods of intensified training in adolescent athletes.

Circulating Levels of Bone Markers after Short-Term Intense Training with Increased Dairy Consumption in Adolescent Female Athletes.

Thirteen female adolescent soccer players (14.3 ± 1.3 years) participated in a cross-over, double-blind trial examining the effects of Greek yogurt (GY) consumption on bone biomarkers during 5 days of intense soccer training. The study took place over two intervention weeks, which consisted of a pre-training assessment day, 5 training days, and a post-training assessment day. Participants completed the GY condition and a carbohydrate isocaloric placebo control pudding condition (CHO) in random order, 4 weeks apart. Morning, fasted, resting blood samples were collected pre- and post-training in each condition. Total osteocalcin (tOC), undercarboxylated osteocalcin (unOC), C-terminal telopeptide of type 1 collagen (CTX), osteoprotegerin (OPG), and receptor activator nuclear factor kappa-ß ligand (RANKL) were measured in serum. The results showed no effects for time (pre- to post-training) or condition, and no interaction for tOC, CTX, OPG, RANKL, and the OPG/RANKL ratio. A time-by-condition interaction (p = 0.011) was observed in unOC, reflecting a post-training decrease in the GY, but not the CHO condition (-26% vs. -3%, respectively). However, relative unOC (% of tOC) decreased post-training (-16%), with no differences between conditions. These findings suggest that short-term high-impact intense training had no direct catabolic impact on bone metabolism, with GY adding no benefit beyond that of the isocaloric CHO control pudding.

Cytokine concentrations in saliva vs. plasma at rest and in response to intense exercise in adolescent athletes.

BACKGROUND: Salivary measures are advantageous in conducting large paediatric studies involving repeated measures. However, research measuring salivary cytokines in youth is limited. AIM: Compare salivary with plasma concentrations of inflammatory cytokines at rest and following exercise in adolescent swimmers (21 male, 22 female). METHODS: Following collection of resting saliva and blood samples, participants performed a bout of high-intensity interval swimming, with samples taken again ∼15 min post-swimming and analysed for interleukin-6 (IL-6), interleukin 10 (IL-10), and tumour necrosis factor-alpha (TNF-α). RESULTS: Resting IL-10 was significantly lower, while IL-6 and TNF-α were significantly higher in saliva compared with plasma. IL-10 increased from pre- to post-swimming in plasma, but less so in saliva (51% vs. 29%; p = 0.02). TNF-α decreased post-swimming in saliva, but not in plasma (-27% vs -1%; p = 0.01). IL-6 decreased post-swimming in saliva compared with plasma (-21% vs. -3%; p = 0.06). Intraclass correlation coefficients (ICC) revealed no association between salivary and plasma IL-6 and TNF-α, while IL-10 showed a weak correlation only at rest (ICC = 0.39; p = 0.05). CONCLUSIONS: Differences in concentrations and exercise responses, along with weak correlations, suggest that salivary cytokine levels are not an accurate representation of blood cytokine levels, and should not be used as a surrogate measure in paediatric studies.

Changes in Inflammatory Cytokines and Irisin in Response to High Intensity Swimming in Adolescent versus Adult Male Swimmers.

Swimming is a popular youth sport that is considered beneficial for cardiovascular fitness. However, the potential inflammatory outcomes of high intensity swimming in younger swimmers are unclear, as is the response of irisin, a myokine released during exercise with anti-inflammatory properties. This study compared the plasma concentrations of interleukins 1-beta (IL-1ß), 6 (IL-6), 10 (IL-10), tumor necrosis factor alpha (TNF-α) and irisin in response to intense swimming between adolescent and adult male swimmers. Thirty-two swimmers (16 adolescents, 14 ± 1 years; 16 adults, 21.5 ± 3.1 years) completed a high intensity interval swimming trial. At rest, only TNF-α was higher (33%, p < 0.05) in adolescents compared with adults. There was an overall significant increase in IL-1ß from pre- to post-swimming (3% in adolescents, 24% in adults), but no significant interaction. IL-10 significantly increased in both groups (+34% in adolescents, +56% in adults). IL-6 and TNF-α increased significantly (+32% and +26%, respectively) in adults, but not in adolescents (+2% and -9%, respectively). Adults showed a small, but significant decrease in irisin (-5%), with no change in adolescents. The lack of an IL-6, TNF-α and irisin response to intense swimming in adolescent swimmers may suggest a blunted inflammatory and myokine response following high intensity exercise in trained youth.

Effects of Post-Exercise Whey Protein Consumption on Recovery Indices in Adolescent Swimmers.

Purpose: This study examined the effect of whey protein consumption following high-intensity interval swimming (HIIS) on muscle damage, inflammatory cytokines and performance in adolescent swimmers. Methods: Fifty-four swimmers (11-17 years-old) were stratified by age, sex and body mass to a whey protein (PRO), isoenergetic carbohydrate (CHO) or a water/placebo (H2O) group. Following baseline blood samples (06:00 h) and a standardised breakfast, participants performed a maximal 200 m swim, followed by HIIS. A total of two post-exercise boluses were consumed following HIIS and ~5 h post-baseline. Blood and 200 m performance measurements were repeated at 5 h, 8 h and 24 h from baseline. Muscle soreness was assessed at 24 h. Creatine kinase (CK), interleukin-6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor-alpha (TNF-α) were measured in plasma. Results: No difference in 200 m swim performance was observed between groups. CK activity was elevated at 5 h compared to baseline and 24 h and at 8 h compared to all other timepoints, with no differences between groups. Muscle soreness was lower in PRO compared to H2O (p = 0.04). Anti-inflammatory IL-10 increased at 8 h in PRO, while it decreased in CHO and H2O. Conclusions: Post-exercise consumption of whey protein appears to have no additional benefit on recovery indices following HIIS compared to isoenergetic amounts of carbohydrate in adolescent swimmers. However, it may assist with the acute-inflammatory response.

Effects of post exercise protein supplementation on markers of bone turnover in adolescent swimmers.

BACKGROUND: This study examined the effects of whey protein supplementation, compared with an isocaloric carbohydrate beverage and water, consumed immediately following an intense swimming trial on bone turnover in adolescent swimmers. METHODS: Fifty-eight (31 female, 27 male) swimmers (14.1 ± 0.4 years) were stratified into three groups matched for age, sex and body mass. The protein and carbohydrate groups consumed two isocaloric post-exercise beverages each containing 0.3 g.kg- 1 of whey protein (with ~ 6 mg of calcium) or maltodextrin while the control group consumed water. Participants provided a morning, fasted, resting blood sample, then performed an intense swimming trial consisting of a maximal 200 m swim followed by a high intensity interval swimming protocol (5x100m, 5x50m and 5x25m; 1:1 work-to-rest ratio). Following swimming, they consumed their first respective post-exercise beverage, and 2 h later, they performed a second maximal swim immediately followed by the second beverage. Approximately 3 h after the second beverage, two post-consumption blood samples were collected at 8 h and 24 h from baseline. Procollagen type 1 intact N-terminal propeptide (PINP) and carboxy-terminal collagen crosslinks (CTXI) were measured in serum. The multiples of medians of PINP and CTXI were also used to calculate bone turnover rate and balance. RESULTS: No significant changes were observed in PINP. CTXI increased (+ 11%) at 8 h in all groups, but then significantly decreased (- 22%) at 24 h in the protein group only. The protein group also had a significantly higher calculated rate of bone turnover at 8 h and 24 h compared to baseline, which was not observed in the other groups. CONCLUSIONS: These results shed light on the potential importance of protein consumed shortly after intense swimming in promoting positive bone turnover responses up to 24 h following exercise in adolescent athletes. CLINICAL TRIAL REGISTRATION: ClinicalTrials.gov PRS; NCT04114045. Registered 1 October 2019 - Retrospectively registered.

Effects of Plyometric and Resistance Training on Muscle Strength, Explosiveness, and Neuromuscular Function in Young Adolescent Soccer Players.

McKinlay, BJ, Wallace, P, Dotan, R, Long, D, Tokuno, C, Gabriel, D, and Falk, B. Effects of plyometric and resistance training on muscle strength, explosiveness, and neuromuscular function in young adolescent soccer players. J Strength Cond Res 32(11): 3039-3050, 2018-This study examined the effect of 8 weeks of free-weight resistance training (RT) and plyometric (PLYO) training on maximal strength, explosiveness, and jump performance compared with no added training (CON), in young male soccer players. Forty-one 11- to 13-year-old soccer players were divided into 3 groups (RT, PLYO, and CON). All participants completed isometric and dynamic (240°·s) knee extensions before and after training. Peak torque (pT), peak rate of torque development (pRTD), electromechanical delay (EMD), rate of muscle activation (Q50), m. vastus lateralis thickness (VLT), and jump performance were examined. Peak torque, pRTD, and jump performance significantly improved in both training groups. Training resulted in significant (p ≤ 0.05) increases in isometric pT (23.4 vs. 15.8%) and pRTD (15.0 vs. 17.6%), in RT and PLYO, respectively. During dynamic contractions, training resulted in significant increases in pT (12.4 and 10.8% in RT and PLYO, respectively), but not in pRTD. Jump performance increased in both training groups (RT = 10.0% and PLYO = 16.2%), with only PLYO significantly different from CON. Training resulted in significant increases in VLT (RT = 6.7% and PLYO = 8.1%). There were no significant EMD changes. In conclusion, 8-week free-weight resistance and plyometric training resulted in significant improvements in muscle strength and jump performance. Training resulted in similar muscle hypertrophy in the 2 training modes, with no clear differences in muscle performance. Plyometric training was more effective in improving jump performance, whereas free-weight RT was more advantageous in improving peak torque, where the stretch reflex was not involved.

Effects of Motivational Self-Talk on Endurance and Cognitive Performance in the Heat.

PURPOSE: We tested the effectiveness of a 2-wk motivational self-talk (MST) intervention-specific to heat tolerance-on endurance capacity and cognitive function in the heat. METHODS: Eighteen trained male (n = 14) and female (n = 4) cyclists randomly received 2 wk of MST training (n = 9) or a control regimen (CON, n = 9). The experimental protocol was a PRE/POST design consisting of 30 min of cycling at 60% peak power output (PPO) in the heat (35°C, 50% relative humidity, ~3.0 m·s airflow), a 30-min rest period, followed by a time to exhaustion (TTE) test at 80% PPO, and an identical rest period. Executive function, reaction time, and working memory were tested at baseline and each rest period. Key measures included TTE, speed and accuracy on the cognitive tests, rectal temperature, HR, oxygen consumption, and RPE. RESULTS: Group (MST vs CON) × test (PRE vs POST) × time repeated-measures ANOVA revealed that MST significantly increased TTE from PRE (487 ± 173 s) to POST (679 ± 251 s, P = 0.021) concurrent with a higher terminating rectal temperature (PRE, 38.5°C ± 0.2°C; POST, 38.8°C ± 0.4°C; P = 0.023); no TTE (PRE, 531 ± 178 s; POST, 510 ± 216 s; P = 0.28) or rectal temperature (PRE, 38.4°C ± 0.3°C; POST, 38.4°C ± 0.2°C; P = 1.000) changes were found in CON. MST significantly improved both speed and accuracy for executive function from PRE/POST, with no PRE/POST differences for CON on any cognitive measure. There were no interactions (all P > 0.05) for other key measures. CONCLUSION: Motivational self-talk is effective in altering the internal psychophysiological control of exercise and plays a role in improving endurance capacity and executive function in the heat.

Salivary cortisol and testosterone responses to resistance and plyometric exercise in 12- to 14-year-old boys.

This study examined changes in salivary testosterone and cortisol following resistance and plyometric exercise protocols in active boys. In a crossover experimental design, 26 peri-pubertal (12- to 14-year-old) soccer players performed 2 exercise trials in random order, on separate evenings, 1 week apart. Each trial included a 30 min control session followed by 30 min of either resistance or plyometric exercise. Saliva was collected at baseline, post-control (i.e., pre-exercise), and 5 and 30 min post-exercise. There were no significant differences in the baseline hormone concentrations between trials or between weeks (p > 0.05). A significant effect for time was found for testosterone (p = 0.02, [Formula: see text] = 0.14), which increased from pre-exercise to 5 min post-exercise in both the resistance (27% ± 5%) and plyometric (12% ± 6%) protocols. Cortisol decreased to a similar extent in both trials (p = 0.009, [Formula: see text] = 0.19) from baseline to post-control and then to 5 min post-exercise, following its typical circadian decrease in the evening hours. However, a significant protocol-by-time interaction was observed for cortisol, which increased 30 min after the plyometrics (+31% ± 12%) but continued to decrease following the resistance protocol (-21% ± 5%). Our results suggest that in young male athletes, multiple modes of exercise can lead to a transient anabolic state, thus maximizing the beneficial effects on growth and development, when exercise is performed in the evening hours.