United Kingdom

Background: Association of dietary intake and serum level of polyunsaturated-fatty-acids (omega-3 PUFA) with physical function and body composition in older adults remained inconclusive. Method: Study subjects were 554 women from the Osteoporosis-Risk-Factor-and-PreventionFracture Prevention-Study, with dietary intake assessed with 3-day food record at baseline. Total omega-3 PUFA and DHA blood concentrations were analysed using a highthroughput proton (1H) NMR spectrometry metabolomics platform at the University of Eastern Finland. Multiple physical function tests and short physical performance battery (SPPB) was calculated at the baseline and 3-year followup. Results: The multivariable adjusted models showed statistically-significant associations for ALA with higher SPPB (β= 0.118, P=0.024), knee-extension-force at baseline (β= 0.075, P=0.037) and lower fat mass (β= - 0.081, P=0.034), as well as longer one-leg-stance (βbaseline= 0.119, βfollw-up=0.090, P&lt;0.023), higher walking speed (βbaseline= 0.113, βfollw-up=0.144, P&lt;0.047), and ability to squat to the ground (βbaseline= 0.110, βfollwup=0.096, P&lt;0.047) at baseline and follow-up. Total omega-3 PUFA was statistically-significantly associated with better SPPB (βbaseline= 0.108, P=0.039), one-leg-stance (βbaseline= 0.102, P=0.041), walking speed (βbaseline= 0.110, βfollw-up=0.128, P&lt;0.028) and ability to squat (βbaseline= 0.110, P=0.028). Associations for dietary EPA and DHA, serum omega-3 PUFA and DHA with physical function and body composition was not significant.

ISTRC 2021 - International Sarcopenia Translational Research Conference

Dietary omega-3 polyunsaturated fatty acid and alpha-linolenic acid are associated with sarcopenia indices in older women 65-72 years.
Masoud Isanejad 1, 2, Toni Rikkonen 3, Behnam Tajik 4, Joonas Sirola

fatty acids

omega 3

sarcopenia

Welcome to the inaugural International Sarcopenia Translational Research Conference.  “Where Worlds Meet” – the event brings together a broad range of researchers and practitioners in a truly interdisciplinary forum, covering discovery science through to clinical practice.

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Background: Association of dietary intake and serum level of polyunsaturated-fatty-acids (omega-3 PUFA) with physical function and body composition in older adults remained inconclusive. Method: Study subjects were 554 women from the Osteoporosis-Risk-Factor-and-PreventionFracture Prevention-Study, with dietary intake assessed with 3-day food record at baseline. Total omega-3 PUFA and DHA blood concentrations were analysed using a highthroughput proton (1H) NMR spectrometry metabolomics platform at the University of Eastern Finland. Multiple physical function tests and short physical performance battery (SPPB) was calculated at the baseline and 3-year followup. Results: The multivariable adjusted models showed statistically-significant associations for ALA with higher SPPB (β= 0.118, P=0.024), knee-extension-force at baseline (β= 0.075, P=0.037) and lower fat mass (β= - 0.081, P=0.034), as well as longer one-leg-stance (βbaseline= 0.119, βfollw-up=0.090, P<0.023), higher walking speed (βbaseline= 0.113, βfollw-up=0.144, P<0.047), and ability to squat to the ground (βbaseline= 0.110, βfollwup=0.096, P<0.047) at baseline and follow-up. Total omega-3 PUFA was statistically-significantly associated with better SPPB (βbaseline= 0.108, P=0.039), one-leg-stance (βbaseline= 0.102, P=0.041), walking speed (βbaseline= 0.110, βfollw-up=0.128, P<0.028) and ability to squat (βbaseline= 0.110, P=0.028). Associations for dietary EPA and DHA, serum omega-3 PUFA and DHA with physical function and body composition was not significant.

poster

Background: Sarcopenia refers to the progressive loss of skeletal muscle mass and function, generally associated with ageing (1). There is growing evidence that sarcopenia is more common in people with cognitive impairment (2). Dementia is the term applied to the clinical syndrome of progressive cognitive decline, negatively impacting on daily function (3). A diagnosis of dementia relies on history taking, examination, and limited investigations, including neuroimaging. A challenging aspect of dementia assessment is that disease severity (assessed on neuropathological features) is not always well correlated with clinical severity (4). Physical frailty may also influence the clinical severity, whereby frailer people are more likely to have a clinical diagnosis of dementia with the same burden of neuropathology as more robust individuals (5). Sarcopenia, representing the physical component of frailty, may provide a clinical tool with which to risk stratify. In terms of sarcopenia evaluation and assessment of skeletal muscle mass and quality, Magnetic Resonance Imaging (MRI) and Computer Tomography (CT) are considered to be gold standards, and Dual-energy X-ray and Bioelectrical impedance analysis are commonly used instruments. However, the clinical evaluation of sarcopenia remains challenging, and a variety of novel tools are being investigated for validity, convenience, and cost-efficiency. Included amongst these are the evaluation of a number of different muscles on CT or MRI, including, recently, the temporalis muscle, as visualised on routine imaging of the brain, as a surrogate marker of sarcopenia evaluation (2). For example, Leitner et al. demonstrated that TMT was associated with Skeletal Muscle Index, suggesting that TMT was reflective of generalised sarcopenia in patients with newly diagnosed brain metastases (6). However, there are currently no studies focussing on patients with memory complaints. Methods: The study cohort comprised of 111 patients attending the memory clinic in 2014. The inclusion criteria were as follows: (a) available coronal MRI of the brain of adequate quality for morphometric assessment, and (b) Frailty Index (FI) data available. FI measures frailty as a ratio between health deficits present in the individual, and the total number of health deficits measured, which is constructed following the guidelines by Rockwood et al (7). Frailty was defined as FI ≥ 0.25, as previously published (7). TMT was analysed on coronal, T1-weighted MR images. The orbital roof and Sylvian fissure were utilised as anatomical landmarks. Measurements were taken perpendicular to the long axis of the muscle. TMT was measured bilaterally, and the average was calculated, resulting in one mean TMT per patient. Shapiro-Wilk test was utilised to assess for normality of distribution of muscle thickness across the cohort. Spearmen correlation test was used to investigate the relationship between TMT and FI. Mann-Whitney U test was utilized to compare distribution of TMT between frail and non-frail patients. Results: Assessment by Shapiro-Wilk test (p = 0.002) suggested a low probability of normal distribution of TMT measures. Spearman correlation test was thus used, and a significant negative correlation of modest magnitude was demonstrated between average TMT and frailty as assessed using FI (rho = -0.24, p = 0.01). Mann-Whitney U test revealed significant difference (Z = -3.341, p = 0.001) in the distribution pattern of average TMT between non-frail (median = 5.7mm, interquartile range = 4.8-6.8) and frail (4.5mm, IQR =4.0-5.6). Together, this supports the hypothesis that increased TMT is associated with decreased frailty in this cohort, and may act as a radiological biomarker of sarcopenia. This is in agreement with previously published studies in other patient groups. This is the first study to examine the use of TMT measurement on coronal MRI in patients with memory complaints; since this is a routine investigation in this cohort, the measurement of TMT may, in future, represent a convenient and fast method by which sarcopenia and frailty can be assessed objectively. Future studies might explore the relationship between TMT and long-term clinical outcomes. Conclusions: The findings of this study support the published literature, and suggest that temporalis muscle thickness, as measured on MRI, may act as a radiological biomarker of sarcopenia in a memory clinic cohort.

Identifying Incidental Biomarkers of Sarcopenia from Routine Imaging in a Memory Clinic Cohort

Background: • Strength and balance training can improve mobility and decrease fall risk in older adults. • But less 5% of older adults, 65 years and older, get enough strength and balance training. • Initiating strength training when protein intake in insufficient can limit gains in muscle strength and cause weight loss. • The biggest challenge is in developing real-world, sustainable models on balance and strength training for older adults with chronic conditions. Methods: • The study is an 8-week, assessor blinded, multisite, stepped wedged, pilot randomized controlled trial. • Participants enrolled in the study received: i. A single one-on-one session with the exercise physiologist; ii. 16 exercise physiologist-led, 1-hour group sessions on balance and functional strength training; iii.2 dietitian-led, 1-hour group sessions on protein intake. • There were 3 urban sites and 1 rural site (Kinnect to Wellness). • We analyzed secondary outcomes using exploratory analyses. Main findings: • The MoveStrong program was feasible in terms of recruitment and adherence. 

Encouraging pre-frail older adults to "MoveStrong": A pilot randomized controlled trial

Background: Visual impairment is common in older adults. The most common causes of age-related visual impairment are presbyopia, cataracts, macular degeneration, and glaucoma, which affect visual acuity. Only two studies have shown that visual impairment can be a predictor of low muscle mass and low muscle strength, indicating a potential correlation between visual impairment and sarcopenia. Methods: Older adults were recruited from the community, and their sarcopenia status was identified using the SARCF questionnaire. Visual acuity was assessed through smartphone-based visual acuity measurement (Smart Optometry) (Figure 1), and values were presented in decimal form. The visual acuity level was categorised into normal visual function (≥0.8), mild visual loss (0.3–0.8), and moderate to severe visual loss (<0.3). Pearson correlation test was performed to measure the association between the SARC-F score and visual acuity. An analysis of variance with a post hoc Bonferroni test was conducted to compare SARC-F scores among different visual acuity levels. This study was approved by the Taipei Medical University Joint Institutional Review Board (N202002009) and registered on clinical.trials.gov (NCT04702828). Results: In total, 100 community-dwelling older adults were recruited. A significant association was noted between SARC-F score and visual acuity (p < 0.001, r = −0.348). Old people with moderate to severe visual loss had significantly higher SARC-F scores relative to those with normal visual function (p = 0.049, 95% confidence interval [CI] = −4.695 to −0.011) and mild vision loss (p = 0.025, 95% CI = −2.379 to −0.117). Conclusions: Sarcopenia is associated with vision loss in older adults. Only when visual acuity reaches a moderate level does it exert a large effect on sarcopenia. This may be due to the reduction of muscle function in visually impaired older adults who restrict themselves from engaging in physical activities. The improvement of visual impairment, such as through prescription glasses or surgical intervention, are required to prevent sarcopenia. Older adults with visual impairment should be advised to undergo sarcopenia assessment.

Sarcopenia and Vision Loss in Community-Dwelling Older Adults

Introduction: 1) Dietary intake and nutritional status is a key factor in the outcomes of older individuals. Changes in dietary habits, and nutritional intake contribute considerably to the progression of frailty and sarcopenia. 2) Robust and highly acceptable home urine collection can provide objective dietary information, with minimal burden and can be used to detect shifts in dietary intake in individuals who may be transitioning into a pre-frail or frail state. Methods: 1) 38 individuals aged over 60, collected three rst morning void spot urine samples over the course of a week, immediately after attending physical capability screening. Dietary intake biomarkers were measured using Liquid Chromatography (LC) Mass Spectrometry (MS). 2) Participants completed urine sampling and physical screening again, after 12-weeks to validate observations made at baseline. Results: Higher concentrations of protein exposure biomarkers were associated with positive outcomes in the various physical capability tests. 1-Methylhistidine (1-MH) and 3- Methylhistidine (3-MH) are both biomakers of poultry and high quality red meat intake, and were signicantly associated with the 5-Chair Stand Test; 1-MH (r = -0.55, P < 0.001), 3-MH (r = -0.38, P = 0.012). Categorisation of grip strength (Low; < 22 Kg in Females and < 32 Kg in Males), showed elevated excretion of the dietary protein biomarker taurine (P = 0.03) and para-Cresol Glucuronide (P = 0.04), which is produced following intestinal catabolism of dietary tyrosine. Conclusions: This pilot study has demonstrated the feasibility of home urine collection in a group of older individuals. On-going studies are using this urine sampling protocol for undernutrition screening in the community and investigating the link between nutritional status and surgical outcomes. 

Biomarkers of dietary intake and their association with physical capability assessments

Introduction: Sarcopenia is defined as the presence of both decreased muscle mass and reduced muscle function. It is highly prevalent amongst the elderly. Current treatment approaches involve exercise and dietary programmes. The RANK-RANKL pathway, a major regulator of bone mass, is inhibited by endogenous osteoprotegerin (OPG) and therapeutic Denosumab. Hypothesis: RANK-RANKL inhibition exerts beneficial effects on muscle mass or function. Study objectives: To determine the effect of OPG on myogenesis and muscle atrophy/hypertrophy in primary myoblasts in vitro. Results: Cytotoxicity of hOPG on primary mouse myoblasts/tubes - MTT was used to measure the cytotoxicity of hOPG on primary mouse myoblasts/tubes - No significant changes in viability were observed in treated proliferating and differentiating myoblasts (data not included here). - OPG treated to myotubes significantly increased their viability. The effect of hOPG on myotube phenotype - MF20 immuno-histochemical staining was used to investigate myotube phenotype when subject to hOPG treatment. - hOPG significantly increased fusion index when treated to differentiating myoblasts for 5 days, and significantly increased, in a dose dependent manner, all phenotype parameters when treated to myotubes. Conclusions: Summary of findings • hOPG (5-20 ng/ml) is not cytotoxic to myoblasts, differentiating myoblasts and myotubes. • hOPG significantly improves the viability of myotubes • Myotube diameter and fusion index is significantly improved when hOPG 5 ng/ml is treated to differentiating myoblasts • hOPG significantly improves myotube phenotype (dose dependent) when treated at late stages of myogenesis • Inhibition of RANKL may influence muscle.

Can RANKL inhibition ameliorate sarcopenia? An in vitro study.

Background: Periodic paralysis (PP) is a rare genetic muscle disease caused by mutations in skeletal muscle ion channels and characterised by episodic flaccid paralysis in association with high (HyperPP) or low serum potassium. Around age 40 the severity of attacks reduces or attacks may even stop but permanent progressive weakness develops. The reason for this is not known. This phenotype transition coincides with the age at which muscle mass and optimal motor function start to decline in healthy individuals. We sought to determine if the phenotype transition in PP is linked to the normal ageing phenotype transition and to explore the mechanisms involved. Methods: A mouse model of Hyper PP, known as Draggen (I582V in SCN4A), was compared to wild-type littermates across a range of ages (13-104 weeks). These mice have already been shown to have episodic attacks and develop permanent progressive weakness from middle age. Only male mice were used as penetrance is incomplete in females. We adapted the Muscle Velocity Recovery Cycle technique from humans to examine murine muscle excitability in vivo. We then examined changes in potassium-induced weakness or caffeine contracture force with age using ex vivo muscle tension testing. Muscles were further characterised by either western blot, histology, or energy charge measurement. For normally distributed data a student’s t-test (±Welch correction) or One or Two-way ANOVA was performed to determine significance. For data that was not normally distributed, Welch Rank test, Mann Whitney U test or Kruskal Wallis ANOVA was performed. When an ANOVA was significant (P<0.05) post hoc Tukey testing was used. Conclusions: 1. The phenotype transition with age is replicated in a mouse model of PP. 2. Intrinsic muscle ageing protects against potassium-induced weakness in Draggen mice. However, it also appears to accelerate impairment of sarcoplasmic reticulum calcium release, mitochondrial impairment and the development of core-like regions, suggesting acquired RyR1 dysfunction as the potential aetiology. 3. This work provides a first description of mechanisms involved in phenotype transition with age in PP. It also a demonstrates how studying phenotype transition with age in monogenic disease can yield novel insights into both disease physiology and the ageing process itself.

Ageing Contributes to Phenotype Transition in the Draggen Mouse Model of Periodic Paralysis

Background: Late-onset Pompe disease (LOPD) is a rare disorder produced by mutations in the GAA gene characterized by progressive muscle weakness. LOPD muscle biopsies show accumulation of glycogen inside lysosomes along with autophagic vacuoles associated with atrophic muscle fibers. The mechanism leading to muscle fiber atrophy and death are not well known. Methods: We analyzed the expression of several atrogens and their regulatory mechanisms on muscle biopsies (n:7) of LODP patients using RealTime-PCR, WB and IF. We obtained myoblasts from biopsies of LOPD patients and studied their proliferation and differentiation rate. We transfected myoblasts with BNIP3 and studied size of the myotubes. We used non-parametric statistic studies to compare groups. P was considered signigficative if P was lower than 0.05. Conclusion: These findings suggest a cascade with inhibition of the AKT-mTOR pathway, activation of expression of BNIP3 leading to mitochondrial damage, mitophagy, and finally to progressive muscle fiber atrophy. Our results open the door to potential new treatments targeting BNIP3 and reducing its deleterious effects on muscle fiber atrophy in Pompe disease.

BNIP3 is involved in muscle fiber atrophy in Late-Onset Pompe Disease

Why Muscle Ageing? Sarcopenia is the age-associated loss of skeletal muscle strength, mass, and function. Sarcopenia exacts a huge personal, social, and economic toll including the loss of independence, a higher risk of institutionalisation, reduced quality of life, and high public health costs. Current interventions - such as exercise, nutritional supplementation, and HRT - engender blunted and highly variable results, and do not resolve the underlying causes of muscle ageing. As such, and in the face of population ageing, there is great interest in better understanding the underlying mechanisms driving skeletal muscle ageing in the hope of introducing preventative interventions and therapeutics. Why Cellular Senescence? Cells enter a senescent state when exposed to a variety of stressors. This cellular senescence (CS) state is defined by irreversible growth arrest and the secretion of pro-inflammatory cytokines, chemokines, and proteases, which are collectively known as the senescence-associated secretory phenotype (SASP). The SASP is thought to contribute to tissue dysfunction by inducing paracrine CS and inflammation, impairing stem cell function, and altering the composition of the extracellular matrix. Senescent cells (SnCs) amass in different tissues with age and at etiological sites in multiple chronic conditions. Importantly, removal of SnCs has been shown to alleviate several pathologies in mice and humans in the context of ageing and age-related diseases. Despite such advancements, it remains unclear whether CS occurs in skeletal muscle and, if so, what its effects during muscle ageing may be. Aims and methods: Using histological and immunohistochemical staining techniques, I aimed to: • Characterise the senescence signature in mouse muscle in vivo. • Determine the phenotypic consequences of CS in skeletal muscle by eliminating SnCs in mice. • Assess the translational potential of these findings in human muscle. Results: Expression of Senescence Markers in Mouse Muscle Fibres - No single, stand-alone marker can identify a senescent cell. Therefore it is standard in the field to assess the expression of multiple markers simultaneously. Here, I have used immunofluorescence stainings of the senescence markers p16 (A), p21 (B), TAF (C), HMGB1 (D), and Lamin B1 (E) to better characterise the senescence phenotype in young and old myofibres in vivo. The results indicate the development of a senescence phenotype in mouse skeletal muscle with age. Study Design and MorpholoCharacterisation Following Senescent Cell Clearance - (A) Diagram depicting study design and treatment protocol. (B) Representative images portraying haematoxylin and eosin, and masson's trichrome stainings. Blue regions indicate fibrotic areas. (C) Dot plots representing fibre size, and percentage fibrotic area in old vehicle, APtreated and D+Q-treated mice. Results indicate improved muscle morphology with SnC clearance. Heatmap of Correlations between Senescence Markers, Morphology and Inflammatory SASP factors - Relationships of interest have been highlighted with a heavierset border. Dark blue indicates strong positive correlation. Dark red indicates strong negative correlation. Conclusions: • In line with the senescence phenotype, markers of senescence p21, p16, and TAF increase with age, while HMGB1 and Lamin B1 are lost with age in mouse skeletal muscle in vivo. • Pharmacogenetic and pharmacologic removal of senescent cells improves overall muscle morphology, particularly fibre size and fibrosis. This indicates that CS is an important factor in skeletal muscle ageing. • p21 and TAF predict severity of morphological decline and SASP-mediated inflammation observed during mouse muscle ageing. • Consistent with a senescence phenotype p16 is elevated with age in human muscle in vivo. However, HMGB1 and Lamin B1 are relatively unchanged. Partial reproducibility of the senescence signature from mouse to human muscle points to limitations in using model organisms to approximate human expression. • Quantifications of additional senescence markers in human muscle are ongoing. Correlations between expression of human senescence markers and physical function/body composition data are also currently underway.

Investigating the Role of Cellular Senescence in Muscle Ageing

Introduction: Sarcopenia has been defined as a progressive and generalised skeletal muscle disorder that occurs in advanced ageing (1). Sarcopenia is associated with increased mortality, disability, risk of falls and increased cost of care during hospital admissions (1-4). In 2019, European Working Group of Sarcopenia in Older People updated their definition of sarcopenia (EWGSOP2) (5) and placed a greater emphasis on the importance of muscle strength. In the last decade, Ireland has experienced an increase in the number of older adults (>65) and the Central Statistics Office has predicted that, by 2030, one in five Irish people will be over the age of 65 (6). Currently, the prevalence of sarcopenia in older adults in Ireland is not known. In community dwelling older adults, the prevalence of sarcopenia is widely debated, ranging from 0-50% (7). The early identification of sarcopenia in older adults is important for preventing the adverse outcomes associated with the muscle disease. One essential component of the model of care of older adults in Ireland is the day hospital service (8). Those attending a day hospital receive medical care and multidisciplinary intervention, due to a change or decline in their health or function. Methods: A consecutive series of older adults attending the Robert Mayne Day Hospital (RMDH) in St James’s Hospital (SJH) Dublin, Ireland were invited to take part in the study between October 2019 and March 2020. Due to limited resources, not all participants could be prescribed a physical accelerometer. However, every third participant was prescribed a physical accelerometer. All accelerometers were worn on the participant’s non-dominant wrist for a 7-day period. Results: A total of 134 participants were recruited to the study. The participants had a mean age of 81.7 (SD 7.1), with a range from 65-98. Sixty-one percent (N=82) were female. Conclusion: This is the first study to apply the EWGSOP2 criteria to older adults living in the community in an Irish day hospital setting. The findings highlight the feasibility of applying the EWGSOP2 screening process easily and safely in this setting. The method of strength test influenced the prevalence as well as which variables were independently associated with sarcopenia, potentially highlighting different pathophysiological changes between the two types of sarcopenia. To appropriately manage sarcopenia, the assessment of sarcopenia needs be more consistent regarding the use of the EWGSOP2 guidelines, as seen with the assessment of muscle strength in this study. More routine assessment of sarcopenia using the EWGSOP2 criteria and consistent assessment tools, would enable a greater understanding of the factors that influence sarcopenia (i.e. falls or hospitalisation) and changes in sarcopenia after hospitalisation or acute illness.

Prevalence of Sarcopenia and Assocaited Factors in Older Adults Attending a Day Hospital Service in Ireland

Introduction: • Sarcopenia often co-occurs with osteoporosis, however most of the evidence has been derived from cross-sectional studies. • There is very limited evidence on the association of sarcopenia and incident osteoporosis from prospective studies. Methods: • 168,684 participants (48.8% of women) were included in this prospective study. • After a median follow-up of 7.4 years, 6,296 were diagnosed with osteoporosis. • Sarcopenia (both pre-sarcopenia and sarcopenia) and its individual components were defined according to the EWGSOP2 criteria (2019). • Associations with incident osteoporosis were investigated using Cox-proportional hazard models adjusted for potential confounder factors: sociodemographic, lifestyle, health-related factors, and morbidity count. Results: • After adjusting for relevant confounding factors, presarcopenia was associated with 1.3-times higher risk of incident osteoporosis in men but not in women. • Sarcopenia was associated with 1.66-times increased incident osteoporosis risk in women but not in men. • In terms of individual components, low muscle mass followed by slow gait speed were associated with a higher risk of incident osteoporosis in both sexes. Discussion: • Pre-sarcopenic men and sarcopenic women had a higher risk of developing osteoporosis even after adjustment for a large range of potential confounders. • Considering that sarcopenia could be prevented, health interventions to improve physical capability may delay or prevent the onset of osteoporosis.

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Dietary omega-3 polyunsaturated fatty acid and alpha-linolenic acid are associated with sarcopenia indices in older women 65-72 years. Masoud Isanejad 1, 2, Toni Rikkonen 3, Behnam Tajik 4, Joonas Sirola

Marieke Bloemink

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Identifying Incidental Biomarkers of Sarcopenia from Routine Imaging in a Memory Clinic Cohort

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