Leucine and ACE inhibitors as therapies for sarcopenia (LACE trial): study protocol for a randomised controlled trial

Background: Sarcopenia (the age-related loss of muscle mass and function) is a major contributor to loss of mobility, falls, loss of independence, morbidity and mortality in older people. Although resistance training is effective in preventing and reversing sarcopenia, many older people are sedentary and either cannot or do not want to exercise. This trial examines the efficacy of supplementation with the amino acid leucine and/or angiotensin converting enzyme inhibition to potentially improve muscle mass and function in people with sarcopenia. Promising preliminary data exist from small studies for both interventions, but neither has yet been tested in adequately powered randomised trials in patients with sarcopenia. Methods: Leucine and ACE inhibitors in sarcopenia (LACE) is a multicentre, masked, placebo-controlled, 2 × 2 factorial randomised trial evaluating the efficacy of leucine and perindopril (angiotensin converting enzyme inhibitor (ACEi)) in patients with sarcopenia. The trial will recruit 440 patients from primary and secondary care services across the UK. Male and female patients aged 70 years and over with sarcopenia as defined by the European Working Group on Sarcopenia (based on low total skeletal muscle mass on bioimpedance analysis and either low gait speed or low handgrip strength) will be eligible for participation. Participants will be excluded if they have a contraindication to, or are already taking, an ACEi, angiotensin receptor blocker or leucine. The primary clinical outcome for the trial is the between-group difference in the Short Physical Performance Battery score at all points between baseline and 12 months. Secondary outcomes include appendicular muscle mass measured using dual-energy X-ray absorptiometry, muscle strength, activities of daily living, quality of life, activity using pedometer step counts and falls. Participants, clinical teams, outcomes assessors and trial analysts are masked to treatment allocation. A panel of biomarkers including microRNAs, neurohormones, genetic polymorphisms and markers of inflammation relevant to muscle pathophysiology will be measured to explore predictors of response and further elucidate mechanisms underlying sarcopenia. Participants will receive a total of 12 months of either perindopril or placebo and either leucine or placebo. Discussion: The results will provide the first robust test of the overall clinical and cost-effectiveness of these novel therapies for older patients with sarcopenia. Trial registration: ISRCTN, ISRCTN90094835. Registered on 18 February 2015

Corticospinal Inputs to Primate Motoneurons Innervatingthe Forelimb from Two Divisions of Primary Motor Cortexand Area 3a

Previous anatomical work in primates has suggested that only corticospinal axons originating in caudal primary motor cortex (“newM1”) and area 3a make monosynaptic cortico-otoneuronal connections with limb motoneurons.By contrast, the more rostral “old M1” is proposed to control motoneurons disynaptically via spinal interneurons. In six macaque monkeys, we examined the effects from focal stimulation within old and new M1 and area 3a on 135 antidromically identified motoneurons projecting to the upper limb. EPSPs withsegmental latency shorter than 1.2 ms were classified as definitively monosynaptic; these were seen only after stimulation within new M1or at the new M1/3a border (incidence 6.6% and 1.3%, respectively; total n=27). However, most responses had longer latencies. Usingmeasures of the response facilitation after a second stimulus compared with the first, and the reduction in response latency after a third stimulus compared with the first, we classified these late responses as likely mediated by either long-latency monosynaptic (n=108) ornon-monosynaptic linkages (n=108). Both old and new M1 generated putative long-latency monosynaptic and non-monosynaptic effects; the majority of responses from area 3a were non-monosynaptic. Both types of responses from new M1 had significantly greateramplitude than those from old M1. We suggest that slowly conducting corticospinal fibers from old M1 generate weak late monosynaptic effects in motoneurons. These may represent a stage in control of primate motoneurons by the cortex intermediate between disynapticoutput via an interposed interneuron seen in nonprimates and the fast direct monosynaptic connections present in new M1.This work was supported by the Wellcome Trust

Association between objectively measured physical activity and opioid, hypnotic, or anticholinergic medication use in older people – data from the Physical Activity Cohort Scotland study

Background: Centrally acting medications cause cognitive slowing and incoordination, which could reduce older people’s physical activity levels. This association has not been studied previously.Objectives: To examine the association between opioid, hypnotic and anticholinergic medication, and objectively measured physical activity, in a cohort of older people.Methods: We used data from the Physical Activity Cohort Scotland, a representative cohort of community-dwelling older people aged 65 and over who were assessed at baseline and again 2-3 years later. Objective physical activity was measured using Stayhealthy RT3 accelerometers over 7 days. Baseline medication use (opioid use, hypnotic use, modified anticholinergic risk score [mARS]) was obtained from linked, routinely collected community prescribing records. Cross-sectional and longitudinal associations between baseline medication use and both baseline activity and change in activity over time were analysed using unadjusted and adjusted linear regression models.Results: 310 participants were included in the analysis; mean age 77 (SD 7) years. No association was seen between baseline use of any medication class and baseline physical activity levels in unadjusted or adjusted models. For change in activity over time, there was no difference between users and non-users of hypnotics or opioids. Higher anticholinergic burden was associated with a steeper decline in activity over the follow up period (mARS=0: -7051 counts/24h/yr; mARS=1-2 -15942 counts/24h/yr; mARS>=3 -19544 counts/24h/yr; p=0.03) and this remained robust to multiple adjustments.Conclusion: Anticholinergic burden is associated with greater decline in objectively measured physical activity over time in older people, a finding not seen with hypnotic or opioid use

Maternal care predicts facial expression processing in macaques

Facial expressions are common across mammals and are essential for social communication. In humans, a rich early social environment is important for the appropriate development of facial expression processing. Whether other animals are similarly reliant on social input for facial expression development, or have a more fixed system, is unknown. Here, we investigated how maternal care influences facial expression processing skills in rhesus macaques (Macaca mulatta). We conducted three experiments quantifying facial expression processing and examined performance in relation to historical maternal data and across age. Facial expression processing skill was predicted by positive social contact with the mother during infancy and increased with age until adulthood. Our findings provide the first evidence that early social input, specifically maternal care, enhances facial expression processing skills in non-human animals. This challenges the notion that facial expression processing systems are entirely hard-wired and innate and emphasizes the importance of flexibility and responsiveness to local conditions

Genetic polymorphisms in the serotonin, dopamine and opioid pathways influence social attention in rhesus macaques (Macaca mulatta)

Behaviour has a significant heritable component; however, unpicking the variants of interest in the neural circuits and molecular pathways that underpin these has proven difficult. Here, we present a comprehensive analysis of the relationship between known and new candidate genes from identified pathways and key behaviours for survival in 109 adult rhesus macaques (Macaca mulatta). Eight genes involved in emotion were analysed for variation at a total of nine loci. Genetic data were then correlated with cognitive and observational measures of behaviour associated with wellbeing and survival using MCMC-based Bayesian GLMM in R, to account for relatedness within the macaque population. For four loci the variants genotyped were length polymorphisms (SLC6A4 5-hydroxytryptamine transporter length-polymorphic repeat (5-HTTLPR), SLC6A4 STin polymorphism, Tryptophan 5- hydroxylase 2 (TPH2) and Monoamine oxidase A (MAOA)) whilst for the other five (5- hydroxytryptamine receptor 2A (HTR2A), Dopamine Receptor D4 (DRD4), Oxytocin receptor (OXTR), Arginine vasopressin receptor 1A (AVPR1a), Opioid receptor mu(μ) 1 (OPRM1)) SNPs were analysed. STin genotype, DRD4 haplotype and OXTR haplotype were significantly associated with the cognitive and observational measures of behaviour associated with wellbeing and survival. Genotype for 5-HTTLPR, STin and AVPR1a, and haplotype for HTR2A, DRD4 and OXTR were significantly associated with the duration of behaviours including fear and anxiety. Understanding the biological underpinnings of individual variation in negative emotion (e.g., fear and anxiety), together with their impact on social behaviour (e.g., social attention including vigilance for threat) has application for managing primate populations in the wild and captivity, as well as potential translational application for understanding of the genetic basis of emotions in humans

Developing and validating attention bias tools for assessing trait and state affect in animals: A worked example with Macaca mulatta

Attention bias is a new approach to assessing animal affect that has shown promising results in several animal species. It describes a tendency to preferentially attend to emotional compared to neutral cues and is influenced by underlying affect. It is important in the early days of this new field that we develop widely utilisable methods and incorporate lessons from the human literature from which tasks are adapted. This fundamental knowledge is critical to the development of standardised and sensitive tools, and the validation of experimental protocols to ensure best practice. Here, we describe protocols for two preferential-looking attention bias tasks. Study 1 involved a manual task using freely available low-cost materials. Study 2 used an automated task requiring specialist equipment and programming, but presumably less prone to noisy data. Tasks were tested with 109 socially housed rhesus macaques, Macaca mulatta, who had been trained to sit by a target, but received no other training. Tasks involved showing animals emotional face pairs (threat-neutral), and subsequent blind coding of video for duration of looking at either face. Three measures of social attention were examined: time spent looking at the threat face (THR), total time looking at the threat-neutral face pair overall (TL), and attention bias difference score (ABD) calculated as time spent looking at the neutral face subtracted from time spent looking at the threat face. Based on the human literature and early primate work, the influence of five potential confounding factors on attention was assessed: trial number, stimulus ID, previous testing experience, time of day and visual field to which the threat face was presented; as were several life history factors: sex, age, and social rank. Both tasks revealed stable individual differences in baseline social attention (THR and TL: effect sizes = 0.15−0.31; repeatabilities = 0.12−0.26; suggesting sensitivity to trait affect), but not ABD (which may be more sensitive to brief shifts in emotion state). All potential confounding factors had a significant effect on at least one measure of social attention. For a subset of monkeys who took part in both Study 1 and Study 2 several years apart (n = 18), there was significant reproducibility between tasks for all three measures (R = 0.15−0.63), supporting an argument for stable individual differences in baseline attention bias, and validating the two tasks for measuring the same trait. The attention bias method shows promise for further development of standardised protocols with animals. We provide framework and recommendations for future method development

Activin type I receptor polymorphisms and body composition in older individuals with sarcopenia-Analyses from the LACE randomised controlled trial.

BACKGROUND: Ageing is associated with changes in body composition including an overall reduction in muscle mass and a proportionate increase in fat mass. Sarcopenia is characterised by losses in both muscle mass and strength. Body composition and muscle strength are at least in part genetically determined, consequently polymorphisms in pathways important in muscle biology (e.g., the activin/myostatin signalling pathway) are hypothesised to contribute to the development of sarcopenia. METHODS: We compared regional body composition measured by DXA with genotypes for two polymorphisms (rs10783486, minor allele frequency (MAF) = 0.26 and rs2854464, MAF = 0.26) in the activin 1B receptor (ACVR1B) determined by PCR in a cross-sectional analysis of DNA from 110 older individuals with sarcopenia from the LACE trial. RESULTS: Neither muscle mass nor strength showed any significant associations with either genotype in this cohort. Initial analysis of rs10783486 showed that males with the AA/AG genotype were taller than GG males (174±7cm vs 170±5cm, p = 0.023) and had higher arm fat mass, (median higher by 15%, p = 0.008), and leg fat mass (median higher by 14%, p = 0.042). After correcting for height, arm fat mass remained significantly higher (median higher by 4% padj = 0.024). No associations (adjusted or unadjusted) were seen in females. Similar analysis of the rs2854464 allele showed a similar pattern with the presence of the minor allele (GG/AG) being associated with greater height (GG/AG = 174±7 cm vs AA = 170 ±5cm, p = 0.017) and greater arm fat mass (median higher by 16%, p = 0.023). Again, the difference in arm fat remained after correction for height. No similar associations were seen in females analysed alone. CONCLUSION: These data suggest that polymorphic variation in the ACVR1B locus could be associated with body composition in older males. The activin/myostatin pathway might offer a novel potential target to prevent fat accumulation in older individuals

Decomposing Neural Synchrony: Toward an Explanation for Near-Zero Phase-Lag in Cortical Oscillatory Networks

Background: Synchronized oscillation in cortical networks has been suggested as a mechanism for diverse functions ranging from perceptual binding to memory formation to sensorimotor integration. Concomitant with synchronization is the occurrence of near-zero phase-lag often observed between network components. Recent theories have considered the importance of this phenomenon in establishing an effective communication framework among neuronal ensembles. Methodology/Principal Findings: Two factors, among possibly others, can be hypothesized to contribute to the near-zero phase-lag relationship: (1) positively correlated common input with no significant relative time delay and (2) bidirectional interaction. Thus far, no empirical test of these hypotheses has been possible for lack of means to tease apart the specific causes underlying the observed synchrony. In this work simulation examples were first used to illustrate the ideas. A quantitative method that decomposes the statistical interdependence between two cortical areas into a feed-forward, a feed-back and a common-input component was then introduced and applied to test the hypotheses on multichannel local field potential recordings from two behaving monkeys. Conclusion/Significance: The near-zero phase-lag phenomenon is important in the study of large-scale oscillatory networks. A rigorous mathematical theorem is used for the first time to empirically examine the factors that contribute to this phenomenon. Given the critical role that oscillatory activity is likely to play in the regulation of biological processes at al

Association of bradykinin receptor 2 (BDKRB2) variants with physical performance and muscle mass: Findings from the LACE sarcopenia trial

\ua9 2024 Shrestha et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Introduction: Understanding genetic contributors to sarcopenia (age-related loss of muscle strength and mass) is key to finding effective therapies. Variants of the bradykinin receptor 2 (BDKRB2) have been linked to athletic and muscle performance. The rs1799722-9 and rs5810761 T alleles have been shown to be overrepresented in endurance athletes, possibly due to increased transcriptional rates of the receptor. These variants have been rarely studied in older people or people with sarcopenia. Methods: We performed a post hoc sub-study of the Leucine and ACE (LACE) inhibitor trial, which enrolled 145 participants aged ≥70 years with low grip strength and low gait speed. Participants\u27 blood samples were genotyped for rs179972 using TaqMan and rs5810761 by amplification through Hotstar Taq. Genotypes were compared with outcomes of physical performance and body composition measures. Results: Data from 136 individuals were included in the analysis. For rs1799722 the genotype frequency (TT: 17, CC: 48, CT: 71) remained in Hardy-Weinberg Equilibrium (HWE p = 0.248). There was no difference between the genotypes for six-Minute Walk Distance (6MWD) or Short Physical Performance Battery (SPPB). Men with the TT genotype had a significantly greater 6MWD than other genotypes (TT 400m vs CT 310m vs CC 314m, p = 0.027), and greater leg muscle mass (TT 17.59kg vs CT 15.04kg vs CC 15.65kg, p = 0.007). For rs5810761, the genotype frequency (-9-9: 31, +9+9: 43, -9+9: 60) remained in HWE (p = 0.269). The +9+9 genotype was associated with a significant change in SPPB score at 12 months (-9-9 0 vs -9+9 0 vs +9+9-1, p<0.001), suggesting an improvement. In men, the -9- 9 genotype was associated with lower arm fat (-9-9 2.39kg vs -9+9 2.72kg vs +9+9 2.76kg, p = 0.019). Conclusion: In men, the rs1799722 TT genotype was associated with longer 6MWD and greater leg muscle mass, while the rs5810761 -9-9 genotype was associated with lower arm fat mass