A Leucine-enriched Diet Enhances Overload-induced Growth and Suppresses Markers of Protein Degradation in Aged Rat Skeletal Muscle

Introduction: The hypertrophic response to overload in fast-twitch skeletal muscle is impaired in aged humans and rats, and upregulation of protein degradation pathways are hypothesized to be a contributing factor. Muscle growth occurs when protein synthesis is greater than protein degradation. Dietary supplementation of the essential amino acid leucine has been shown to reduce protein degradation in both young and aged skeletal muscle. Specifically, leucine acts in part by attenuating 5\u27-AMP-activated protein kinase (AMPK) activation as well as the translocation of the forkhead box transcription factor 3A (FoxO3, known to promote transcription of mRNAs encoding degradation pathway proteins) to the nucleus. Akt (a promoter of muscle growth) prevents translocation of FoxO3 into the nucleus by phosphorylating FoxO3 phosphorylation at Ser318/321. However, AMPK, inhibits Akt\u27s phosphorylation of FoxO3, allowing it to enter the nucleus and increase transcription of protein degradation pathway genes encoding ubiquitin ligase proteins such as muscle RING finger 1 (MuRF1) and muscle atrophy F-box (MAFbx, or Atrogin-1). During the aging process, AMPK Thr172 phosphorylation (and thus its activation) is increased, purportedly inhibiting gains in muscle mass and strength. Although dietary leucine supplementation has been shown to enhance strength gians in response to resistance training in young humans, the potential for leucine supplementation to enhance overload-induced muscle hypertrophy in aged humans or animal models has not been examined. Thus, the aim of this study was to determine whether dietary leucine supplementation can attenuate markers of protein degradation and rescue hypertrophy during overload in the fast-twitch skeletal muscles of aged rats to levels comparable to their younger counterparts. It was hypothesized that dietary leucine supplementation during 7 days of fast-twitch plantaris muscle overload would enhance plantaris muscle hypertrophy in aged rats to levels observed in young adult rats not receiving leucine. It was also hypothesized that dietary leucine supplementation during the overload period would alter markers of protein degradation (enhance FoxO3 phosphorylation and reduce the levels of AMPK phosphorylation, Atrogin-1 protein content, and MuRF1 protein content) in the overloaded fast-twitch plantaris muscles of the aged rats to levels observed in young adult rats not receiving leucine. Methods: Young adult (8 mo.) and old (33 mo.) male Fisher 344 x Brown Norway F1 Hybrid (FBN) rats underwent a 1-week unilateral overload of the fast-twitch plantaris muscles via tenotomy of the synergistic gastrocnemius muscle. Within each age group, animals were matched for body weight and separated into either a dietary leucine supplementation group (normal rat chow supplemented by an additional 5% leucine content in place of 5% of the carbohydrate content; n = 7/age group) or placebo group (normal rat chow; n = 6/age group). The leucine groups started the leucine-enriched diet 2 days prior to, and throughout, the overload intervention. All animals had ad libitum access to water and chow during the entire experiment; no differences in daily calorie consumption were observed between the placebo vs. leucine groups within each age group. At the end of the overload period, sham-operated and overloaded plantaris muscles were harvested and analyzed via western blotting for the phosphorylations of AMPK and FoxO3 as well as total levels of Atrogin-1 and MuRF1. A 2x2x2 ANOVA with repeated measures was used for analyses of the effects of age, dietary intervention, and overload (the repeated measure) on muscle hypertrophy. A 2x2 ANOVA was used to measure the percent changes in hypertrophy and western blot analyses. Post-hoc comparisons were accomplished via a Fisher\u27s Least Significant Difference test, with statistical significance being set at p ≤ 0.05. Results: Dietary leucine enrichment significantly (p ≤ 0.05) enhanced overload-induced fast-twitch plantaris muscle hypertrophy in old, but not in young adult, animals. A similar effect was also observed in the slow-twitch soleus muscles, but western blotting analyses are only presented for the fast-twitch plantaris muscles. Sham and overloaded plantaris muscle AMPK phosphorylation was significantly higher in aged animals receiving normal chow compared to young adult animals; however, leucine supplementation in old animals reduced this AMPK phosphorylation to levels similar to young adult animals. Compared to placebo, leucine also non-significantly (p = 0.07) enhanced FoxO3 phosphorylation in the overloaded muscles of both young adult and old animals (thus theoretically reducing FoxO3 translocation to the nucleus). Accordingly, leucine also non-significantly (p = 0.07) reversed the overload-induced increase (from a 22.8% increase to a 17.0% decrease) in Atrogin-1 content in aged muscles and non-significantly (p = 0.14) enhanced the overload-induced decrease in MuRF1 content in the muscles of both age groups. Discussion: These novel findings indicate that a leucine-enriched diet may potentially enhance overload-induced growth of aged fast-twitch muscle, in part by suppressing pathways known to stimulate protein degradation. This is in accord with previous findings of leucine’s suppressive effect on protein degradation in both young adult and aged skeletal muscle under resting conditions. The fact that leucine supplementation enhanced overload-induced hypertrophy only in the old (and not the young) animals may reflect the high growth stimulus of the chronic overload model. That is, the balance of protein synthesis/degradation rates under such a large chronic growth stimulus may not be the limiting factor in young animals, in which muscle growth is not impaired (i.e., synthesis/degradation rates may reach futile levels, and another factor such as sarcomere assembly may be limiting). However, the impaired balance of protein synthesis/degradation rates may be the limiting factor to growth in aged muscle, and leucine may correct this imbalance to restore muscle growth to levels observed in young animals

A Leucine-enriched Diet Enhances Overload-induced Growth and Markers of Protein Synthesis in Aged Rat Skeletal Muscle

Introduction: The hypertrophic response to overload in fast-twitch skeletal muscle is impaired in aged humans and rats, and impaired protein synthesis pathway activation is hypothesized to be a contributing factor. Muscle growth occurs when protein synthesis exceeds protein degradation. Dietary supplementation of the essential amino acid leucine has been shown to enhance protein synthesis in both young and aged skeletal muscle. Leucine acts in part by activating mammalian target of rapamycin (mTOR; a key upstream regulator of protein synthesis pathways) as well as by attenuating the activation of 5\u27-AMP-activated protein kinase (AMPK; a negative regulator of mTOR and protein synthesis). During the aging process, AMPK Thr172 phosphorylation (and thus its activation) is increased, purportedly inhibiting gains in muscle mass and strength. Although dietary leucine supplementation has been shown to enhance strength gains in response to resistance training in young humans, the potential for leucine supplementation to enhance overload-induced muscle hypertrophy in aged humans or animal models has not been examined. Thus, the aim of this study was to determine whether dietary leucine supplementation can enhance markers of protein synthesis and rescue hypertrophy in overloaded fast-twitch skeletal muscles of aged rats to levels comparable to their younger counterparts. It was hypothesized that dietary leucine supplementation during 7 days of fast-twitch plantaris muscle overload would enhance plantaris muscle hypertrophy in aged rats to levels observed in young adult rats not receiving leucine. It was also hypothesized that dietary leucine supplementation during the overload period would suppress AMPK phosphorylation and enhance markers of protein synthesis [70 kDa ribosomal protein S6 kinase (p70S6k), ribosomal protein S6 (rpS6), and eukaryotic elongation factor 2 (eEF2)] in the overloaded fast-twitch plantaris muscles of the aged rats to levels observed in young adult rats not receiving leucine. Methods: Young adult (8 mo.) and old (33 mo.) male Fisher 344 x Brown Norway F1 Hybrid (FBN) rats underwent a 1-week unilateral overload of the fast-twitch plantaris muscles via tenotomy of the synergistic gastrocnemius muscle. Within each age group, animals were matched for body weight and separated into either a dietary leucine supplementation group (normal rat chow supplemented by an additional 5% leucine content in place of 5% of the carbohydrate content; n = 7/age group) or placebo group (normal rat chow; n = 6/age group). The leucine groups started the leucine-enriched diet 2 days prior to, and throughout, the overload intervention. All animals had ad libitum access to water and chow during the entire experiment; no differences in daily calorie consumption were observed between the placebo vs. leucine groups within each age group. At the end of the overload period, sham-operated and overloaded plantaris muscles were harvested and analyzed via western blotting for the phosphorylations of AMPK, p70S6k, rpS6, and eEF2. A 2x2x2 ANOVA with repeated measures was used for analyses of the effects of age, dietary intervention, and overload (the repeated measure) on muscle hypertrophy. A 2x2 ANOVA was used to measure the percent changes in hypertrophy and western blot analyses. Post-hoc comparisons were accomplished via a Fisher\u27s Least Significant Difference test, with statistical significance being set at p ≤ 0.05. Results: Dietary leucine enrichment significantly (p ≤ 0.05) enhanced overload-induced fast-twitch plantaris muscle hypertrophy in old, but not in young adult, animals. A similar effect was also observed in the slow-twitch soleus muscles, but western blotting analyses are only presented for the fast-twitch plantaris muscles. Sham and overloaded plantaris muscle AMPK phosphorylation (Thr172) was significantly higher in aged animals receiving normal chow compared to young adult animals; however, leucine supplementation in old animals reduced this AMPK phosphorylation to levels similar to young adult animals. Phospho-p70S6k (Thr389) and phospho-rpS6 (Ser235/Ser236) were significantly lower in old vs. young overloaded muscles under placebo conditions, but leucine partially restored both p70S6k and rpS6 phosphorylations in old overloaded muscles to that of young adult overloaded muscles. Overload significantly increased total eEF2 content and decreased inhibitory eEF2 phosphorylation (Thr56; normalized to total eEF2) in young adult muscles regardless of leucine supplementation. Total eEF2 content was unaffected by overload in old placebo muscles, but leucine supplementation in old animals non-significantly (p = 0.09) restored the overload-induced increase in total eEF2 content. Muscle eEF2 phosphorylation was unaffected by overload or leucine supplementation in old animals. Discussion: These novel findings indicate that a leucine-enriched diet may potentially enhance overload-induced growth of aged fast-twitch muscle, in part by enhancing pathways known to stimulate protein synthesis. This is in accord with previous findings of leucine’s stimulating effect on protein synthesis in both young adult and aged skeletal muscle under resting conditions. The fact that leucine supplementation enhanced overload-induced hypertrophy only in the old (and not the young) animals may reflect the high growth stimulus of the chronic overload model. That is, the balance of protein synthesis/degradation rates under such a large chronic growth stimulus may not be the limiting factor in young animals, in which muscle growth is not impaired (i.e., synthesis/degradation rates may reach futile levels, and another factor such as sarcomere assembly may be limiting). However, the impaired balance of protein synthesis/degradation rates may be the limiting factor to growth in aged muscle, and leucine may correct this imbalance to restore muscle growth to levels observed in young animals

Differential Eye Movements in Mild Traumatic Brain Injury vs. Normal Controls

Objective measures to diagnose and to monitor improvement of symptoms following mild traumatic brain injury (mTBI) are lacking. Computerized eye tracking has been advocated as a rapid, user friendly and field ready technique to meet this need. Eye tracking data collected via a head mounted, video-based binocular eye tracker was used to examine saccades, fixations and smooth pursuit movement in 60 military Service Members with post concussive syndrome (PCS) and 26 asymptomatic control subjects in an effort to determine if eye movement differences could be found and quantified. The diagnosis of mTBI was confirmed by the study physiatrist’s history, physical examination, and a review of any medical records. Results demonstrated that subjects with symptomatic mTBI had statistically larger position errors, smaller saccadic amplitudes, smaller predicted peak velocities, smaller peak accelerations, and longer durations. Subjects with symptomatic mTBI were also less likely to follow a target movement (less primary saccades). In general, symptomatic mTBI tracked the stepwise moving targets less accurately, revealing possible brain dysfunction. A reliable, standardized protocol that appears to differentiate mTBI from normals was developed for use in future research. This investigation represents a step toward objective identification of those with PCS. Future studies focused on increasing the specificity of eye movement differences in those with PCS are needed

Effects of hyperbaric oxygen on eye tracking abnormalities in males after mild traumatic brain injury

The effects of hyperbaric oxygen (HBO2) on eye movement abnormalities in 60 military servicemembers with at least one mild traumatic brain injury (mTBI) from combat were examined in a single-center, randomized, double-blind, sham-controlled, prospective study at the Naval Medicine Operational Training Center. During the 10 wk of the study, each subject was delivered a series of 40, once a day, hyperbaric chamber compressions at a pressure of 2.0 atmospheres absolute (ATA). At each session, subjects breathed one of three preassigned oxygen fractions (10.5%, 75%, or 100%) for 1 h, resulting in an oxygen exposure equivalent to breathing either surface air, 100% oxygen at 1.5 ATA, or 100% oxygen at 2.0 ATA, respectively. Using a standardized, validated, computerized eye tracking protocol, fixation, saccades, and smooth pursuit eye movements were measured just prior to intervention and immediately postintervention. Between- and within-groups testing of pre- and postintervention means revealed no significant differences on eye movement abnormalities and no significant main effect for HBO2 at either 1.5 ATA or 2.0 ATA equivalent compared with the sham-control. This study demonstrated that neither 1.5 nor 2.0 ATA equivalent HBO2 had an effect on postconcussive eye movement abnormalities after mTBI when compared with a sham-control

A 2-terminal perovskite/silicon multijunction solar cell enabled by a silicon tunnel junction

With the advent of efficient high-bandgap metal-halide perovskite photovoltaics, an opportunity exists to make perovskite/silicon tandem solar cells. We fabricate a monolithic tandem by developing a silicon-based interband tunnel junction that facilitates majority-carrier charge recombination between the perovskite and silicon sub-cells. We demonstrate a 1 cm[superscript 2] 2-terminal monolithic perovskite/silicon multijunction solar cell with a V [subscript OC] as high as 1.65 V. We achieve a stable 13.7% power conversion efficiency with the perovskite as the current-limiting sub-cell, and identify key challenges for this device architecture to reach efficiencies over 25%.Bay Area Photovoltaic Consortium (Contract DE-EE0004946)United States. Dept. of Energy (Contract DE-EE0006707

Gas turbine combustor

A gas turbine engine has a combustor module including an annular combustor having a liner assembly that defines an annular combustion chamber having a length, L. The liner assembly includes a radially inner liner, a radially outer liner that circumscribes the inner liner, and a bulkhead, having a height, H1, which extends between the respective forward ends of the inner liner and the outer liner. The combustor has an exit height, H3, at the respective aft ends of the inner liner and the outer liner interior. The annular combustor has a ratio H1/H3 having a value less than or equal to 1.7. The annular combustor may also have a ration L/H3 having a value less than or equal to 6.0

Assessing patients’ preferences for gender, age, and experience of their urogynecologic provider

Introduction and hypothesis Understanding patient preferences regarding provider characteristics is an under-explored area in urogynecology. This study aims to describe patient preferences for urogynecologic care, including provider gender, age, experience, and presence of medical trainees. Methods This was a multicenter, cross-sectional, survey-based study assessing patient preferences with a voluntary, self-administered, anonymous questionnaire prior to their first urogynecology consult. A 5-point Likert scale addressing provider gender, age, experience, and presence of trainees was used. Descriptive statistics summarized patient characteristics and provider preferences. Chi-squared (or Fisher’s exact) test was used to test for associations. Results Six hundred fifteen women participated from eight sites including all geographic regions across the US; 70.8% identified as white with mean age of 58.5 ± 14.2 years. Urinary incontinence was the most commonly reported symptom (45.9%); 51.4% saw a female provider. The majority of patients saw a provider 45–60 years old (42.8%) with > 15 years’ experience (60.9%). Sixty-five percent of patients preferred a female provider; 10% preferred a male provider. Sixteen percent preferred a provider 60 years old. Most patients preferred a provider with 5–15 or > 15 years’ experience (49% and 46%, respectively). Eleven percent preferred the presence of trainees while 24% preferred trainee absence. Conclusion Patient preferences regarding urogynecologic providers included female gender and provider age 45–60 years old with > 5 years’ experience. Further study is needed to identify qualitative components associated with these preferences