Frontiers in Aging

Fibro-adipogenic progenitors (FAPs) have emerged as central regulators of the skeletal muscle homeostasis and the muscle microenvironment. However, the role of FAPs in the context of exercise-induced muscle hypertrophy remains largely unexplored. Here, we utilized six weeks of blood flow restricted resistance exercise (BFRRE) to study cellular adaptations within the muscle microenvironment accompanying muscle hypertrophy in healthy older individuals. Using flow cytometry, we characterized global changes of key cell populations within the skeletal muscle microenvironment, including FAPs, muscle stem cells (MuSCs), and immune cells. BFRRE induced significant enlargement of both FAPs and MuSCs, consistent with cellular adaptation to exercise. Notably, exercise shifted the FAP pool toward an increased predominance of the CD90high FAP phenotype, without altering total FAP abundance. In addition to matrix and collagen-related genes, transcriptional analysis of genes associated with secretory proteins revealed enrichment of promyogenic factors in CD90high versus CD90low FAPs. Conditioned media experiments of freshly isolated FAPs demonstrated that CD90high FAPs promote myotube growth in vitro compared to CD90low counterparts, suggesting that this phenotypic shift may facilitate muscle hypertrophy. In parallel, BFRRE increased the proportion of pro-inflammatory (CD11c+) macrophages within the skeletal muscle niche, highlighting a dynamic immune response during adaptation. Finally, we show identified possible link between pro-inflammatory macrophages and FAPs, as TNF markedly reduced the proliferation of human primary FAPs ex vivo, suggesting that macrophage-derived signals may attenuate excessive FAP expansion during tissue remodelling. Together, these findings provide new insight into how the remodelling of the cellular niche may support muscle hypertrophy in response to exercise. The coordinated expansion and phenotypic remodeling of FAP and immune cell populations may represent an important mechanism through which exercise supports hypertrophy in older individuals.

Aged skeletal muscle has decreased ability to rebound from physiological stressors such as disuse atrophy. We previously observed that older adults during recovery following muscle disuse were characterized by rapid skeletal muscle immune cell expansion, cellular senescence and collagen deposition, responses that were biased toward older women. Here we used single-nucleus RNA sequencing and complementary in vitro muscle primary cell experiments in young (YF; 22[plusmn]3y; n=8) and older females (OF; 66[plusmn]5y; n=9) to investigate age-mediated cellular function and intercellular communication during recovery from disuse atrophy. Compared with YF, OF exhibited a markedly greater transcriptional response at 7d-recovery (YF: 1,548 DEGs, OF: 7,999 DEGs), driven primarily by slow- (YF: 96 DEGs, OF: 984 DEGs) and fast-twitch myonuclei (YF: 194 DEGs, OF: 1,234 DEGs), satellite cells (YF: 548 DEGs, OF: 2,784 DEGs) and FAPs (YF: 326 DEGs, OF: 2,102 DEGs). Satellite cells from OF demonstrated collagen signatures including elevated THBS1 expression and enrichment of TGF-[beta] signaling. Concurrently, OF FAPs exhibited increased expression of fibroblast activation marker ADAMTS14. Furthermore, NicheNet analyses unmasked altered FAP-satellite cell communication in OF. Complementary In vitro experiments revealed that myogenic progenitor cells collected at 7d-recovery from OF (vs YF) donors displayed impaired myogenic differentiation and a cellular senescence-associated phenotype, while fibroblasts exhibited greater myofibroblast-like activation. Conditioned media derived from OF fibroblasts at 7d-recovery further increased cellular senescence and impaired myogenic potential (vs YF fibroblast conditioned media). Collectively, these findings suggest that recovery from disuse atrophy in older females is characterized by altered FAP-satellite cell communication and intrinsic function which may contribute to poor muscle remodeling.

The effects of older age on the oxidation of energy substrates (carbohydrates, fats) over 24 hr and during activities of daily living (ADL) may have substantial implications for metabolic health. To examine this issue, we evaluated age-related differences in substrate oxidation for 24 hr and periods of sleep, ADLs, and a 30-min treadmill walk (30MTW, 1.3m{middle dot}s^-1) in 17 young (29 {+/-} 5 yr, mean {+/-} SD, 6F) and 16 older (72 {+/-} 5, 8F) healthy active adults. Oxygen consumption (&VdotO2) and carbon dioxide production were measured using a 32,500 L room calorimeter to quantify substrate oxidation, which was normalized to body mass (kJ{middle dot}kg^-1) and total energy expenditure (%). Separately, &VdotO2peak was measured during an incremental treadmill test to exhaustion. &VdotO2peak (ml{middle dot}kg^-1{middle dot}min^-1) was lower in older adults compared with young adults. Older adults oxidized fewer carbohydrates than young over 24 hr (kJ{middle dot}kg^-1, %), sleep (kJ{middle dot}kg^-1), and ADLs (kJ{middle dot}kg^-1, %); and fat oxidation was greater in older than young over 24 hr (kJ{middle dot}kg^-1, %) and ADLs (kJ{middle dot}kg^-1, %). Carbohydrate oxidation did not differ by group during the 30MTW (kJ{middle dot}kg^-1, %), but fat oxidation was greater in older (kJ{middle dot}kg^-1) despite their walking at a greater relative intensity. These results indicate that older adults derived oxidative energy differently than young over 24 hr and during daily tasks. The lower carbohydrate and greater fat contribution to daily energy requirements may be meaningful in the development of metabolic disease in older adults.

Simulated microgravity provides a controlled perturbation for testing how loss of mechanical load affects human tissues. After 24 hours in rotating-wall vessels, neural, cardiac and tonsil organoids developed tissue-specific transcriptional responses that overlapped published spaceflight datasets, accompanied by higher transcriptomic-age estimates in neural and cardiac models, altered cardiac-organoid contraction dynamics and modified tonsil-organoid antiviral gene-expression signatures. These findings establish a multi-organoid framework for studying acute responses to gravitational unloading.

Protein aggregates are a diverse array of intra- and extracellular structures containing misfolded proteins with limited macromolecular movement. Protein aggregation is induced by aging and cellular stress and is a hallmark of neurodegenerative diseases including Huntington's, Alzheimer's, and Parkinson's. Fluorescent tools to visualize global protein aggregation in whole organisms are, therefore, important for screening genes and compounds that may reduce or alter protein aggregation in connection with improved health outcomes. Fluorescent probes that report low fluidity structures in cells, such as molecular rotor dyes, are gaining popularity and overcome certain limitations of targeted markers such as tagged proteins or antibodies. However, permeability has been a key barrier preventing widespread use of such probes in whole organisms, including in C. elegans. To achieve visualization of non-specific protein aggregates in whole C. elegans nematodes without the need for strain engineering, we have developed a simple soaking protocol. Our protocol overcomes the permeability limitations of the commercially available molecular rotor dye, Proteostat, enabling widespread labeling of protein aggregates in live and post-fixed worms. Here, we present our novel approach and demonstrate quantification of protein aggregates in multiple tissues to detect differences in protein aggregation due to both genotype and treatment effects. We also outline anticipated future applications for this protocol, including its adaptation for other dyes, as well as limitations and special considerations for its use.

Purpose: Age is the leading factor for age-related macular degeneration (AMD), in which retinal pigmented epithelium (RPE) atrophy is a hallmark. The nuclear factor-erythroid 2-related factor 2 (Nrf2), a major anti-stress transcription factor, decreases in AMD, suggesting bioenergetic stress interacts with age during AMD pathogenesis. We profiled visual function and gene expression of the retina and RPE-choroid with Nrf2 deletion and aging. Methods: We tested 3mo and 14mo Nrf2 null (Nrf2 KO) and Nrf2 WT mice for optokinetic tracking (OKT) and dark-adapted electroretinography (ERG) responses. Eyes were harvested for retina outer nuclear layer (ONL) thickness analysis and retina and RPE-choroid cup RNA isolation for RNAseq profiling. Results: Visual acuity was reduced with aging with lower values in the Nrf2 KO vs Nrf2 WT comparison at 14mo. We observed aged-related reduction of a-, b-, and c-wave ERG amplitudes in both WT and KO mice. Age-related c-wave decline was exacerbated by Nrf2 loss. By RNAseq, two-way ANOVA of RPE-choroid samples revealed differentially expressed genes (DEGs) with age and genotype main effects, as well as DEGs with age and genotype interaction effects. Age DEGs were over-represented in pathways related to inflammation and phagocytosis. Genotype DEGs were associated with alterations in antioxidant and inflammatory processes. Retina RNAseq showed age and genotype DEGs. Conclusions: Ablation of Nrf2 exacerbates age-related declines suggesting that decrease in Nrf2 with aging has functional consequences for inflammation/resiliency. We provide retina and RPE-choroid bulk-RNAseq datasets from the same cohort of mice that can be interrogated to better understand degeneration of retina/RPE-choroid.

The progressive collapse of protein homeostasis is a defining feature of aging, yet upstream events deteriorating protein synthesis remain poorly defined. Zinc is an essential micronutrient and structural cofactor, but its role in the aging ribosomal proteome is poorly understood. Here, using Caenorhabditis elegans, we show that the whole-body zinc-to-sulfur ratio declines by ~90% between day 1 and day 13 of adulthood. Supplementing aged animals with a low dose of ZnCl2 (10 M) restored organismal zinc levels and improved healthspan, thermotolerance, and oxidative stress resistance, whereas higher doses were toxic. Quantitative proteomics revealed that structural constituents of the ribosome were the most enriched functional category altered by zinc, accompanied by robust induction of metallothionein MTL-1. Zinc-supplemented animals exhibited increased protein synthesis and a transient increase in ribosomal proteins, while 45S pre-rRNA levels remained unaffected, indicating that zinc acts on ribosome structure rather than biogenesis. Zinc supplementation coordinately reversed the age-associated decline of antioxidant, chaperone, proteasome, and energy-metabolism factors. Furthermore, zinc conferred protection in two proteotoxicity models, suppressing polyglutamine aggregation and delaying amyloid-beta-induced paralysis. Our findings establish that zinc preserves translational proteostasis, identifying late-life zinc supplementation as a translatable micronutrient intervention against age-related proteinopathies.

Aging reduces skeletal muscle regenerative capacity, but how the aged environment impairs muscle stem cells (MuSCs) remains unclear. Skeletal muscle acts as a secretory organ, yet age-related changes in its secretome and impact on stem cells are poorly understood. We performed proteomic and lipidomic analyses of conditioned media from primary muscle cells of young (15-25 years) and aged (60-86 years) donors. We identified 127 age-dependent secreted proteins, revealing major remodeling. The lysosomal enzyme acid sphingomyelinase (ASM), which converts sphingomyelin into ceramide, was the most upregulated factor. Recombinant ASM reduced PAX7+ MuSCs in young cultures, showing extracellular ASM impairs stem cell maintenance. Lipidomics revealed disruption of the sphingomyelin-ceramide-S1P axis, with decreased sphingomyelin and S1P, a lipid promoting MuSC proliferation. Exogenous S1P restored the stem cell pool in aged cultures. These findings link secretome remodeling and sphingolipid metabolism to MuSC decline and identify ASM and S1P as targets

Periods of limb immobilization are common in clinical care and lead to rapid losses of muscle mass and strength. We therefore examined whether prehabilitative exercise improves outcomes after disuse by increasing pre-immobilization muscle reserve or by attenuating the biological response to immobilization. Here, we report the primary and muscle-focused secondary outcomes of the PRE-EX (PREhabilitative EXercise) randomized controlled trial (ClinicalTrials.gov: NCT06205784). Fifty-four healthy younger and older women completed four weeks of supervised resistance exercise or no training before eight days of unilateral limb immobilization, followed by four weeks of rehabilitation. From baseline to pre-immobilization, prehabilitation increased quadriceps cross-sectional area (CSA) by 6.7%, maximal voluntary contraction (MVC) by 15.2%, and muscle fiber CSA by 17.7%. Immobilization subsequently reduced quadriceps CSA by 3.2% over 5 days and MVC by ~20%, with similar relative losses in the Prehab and No-train groups. Consequently, quadriceps CSA, the studys primary outcome, remained greater in Prehab than in No-train after immobilization and following rehabilitation, while MVC was greater following rehabilitation. Prehabilitation did not preserve markers of neuromuscular innervation or produce a detectable post-immobilization transcriptomic signature. Instead, immobilization elicited an extensive transcriptional response comprising more than 7000 differentially expressed genes, characterized by activation of inflammatory, proteolytic, stress, and denervation-associated programs together with suppression of mitochondrial and muscle-structural pathways. Network analysis further linked these coordinated molecular programs to disuse-related changes in muscle size, strength, and morphology. The effects of prehabilitation and the subsequent response to immobilization were similar in younger and older women. These findings demonstrate that short-term prehabilitative exercise mitigates the consequences of limb immobilization by establishing greater muscle and strength reserve before disuse, rather than by attenuating the physiological or molecular response to immobilization.

Background: Safely walking through complex environments requires the use of visual information to plan and regulate steps, while managing competing cognitive demands. Age-related changes in visual control of gait and executive function may contribute to unsafe stepping behaviour, particularly under cognitive load. Route previewing may support visual motor planning, but its effects under dual task conditions in older adults are not well understood. Research Question: Does cognitive dual tasking influence gait and gaze behaviour during adaptive walking in older adults, and does previewing the walking route before gait initiation mitigate these effects? Methods: Thirty nine community dwelling older adults (Mage=73.4 years, SD=8.1; 14 male) completed four adaptive walking conditions, combining single and dual-task walking with and without previewing. Per condition, participants completed six trials across different 8-m long coloured walking routes. Gait characteristics were assessed using an instrumented walkway and video-analysis, while gaze behaviour was recorded using mobile eye tracking. Cognitive load was manipulated using a serial subtraction task. Gait and gaze outcomes were evaluated using linear mixed effects models. Results: Dual task performance significantly increased stepping errors, cross-steps, and double limb support, while reducing stride length (p < .001). It also significantly increased the number and duration of task irrelevant fixations (p < .001). After previewing, participants exhibited more efficient gaze behaviour, as evidenced by looking further ahead (larger forward gaze transitions) and fewer irrelevant gaze fixations (p < .002). Significant Task-by-Preview interactions revealed that previewing mitigated the negative effects of dual-tasking on stepping errors and cross steps (p < .001). Significance: Route previewing prior to gait initiation attenuates negative effects of cognitive load on adaptive gait in older adults, likely by supporting visual motor planning. These findings highlight the potential value of simple visual planning interventions for improving adaptive gait in older adults.

Bone loss and metabolic dysregulation often coexist in older adults, yet the relationship between age-related changes in bone cell metabolism and bone-forming activity remains unclear. By modeling both conditions in aged mice and using in vivo stable-isotope tracing, we demonstrated that age-related decline in bone formation was coupled with an intrinsic impairment of glycolysis within bone tissue. Impaired glycolysis was associated with reduced protein levels of key glycolysis enzymes, and transcriptomics analyses revealed compensatory upregulation of glycolysis genes in aged bone. Metabolic intervention through long-term caloric restriction prevented age-related trabecular bone loss. Furthermore, osteoblast-specific genetic enhancement of glycolysis, via overexpression of HIF1 or the rate-limiting enzyme PFKFB3, significantly improved osteoblast function and increased bone mass in aged mice. Together, the study identifies glycolytic decline as a cell-autonomous driver of osteoblast deficiency and provides proof of principle that reactivating glycolysis can promote bone accrual in aging.

The contribution of persistent inward currents (PICs) to spinal motor neurons discharge rate is lower in old compared to young adults. However, whether this difference further progresses at very old age ([&ge;]80 years) and differs between males and females remains unknown. Thirty young (26 {+/-} 4 years, 15 females), thirty old (72 {+/-} 5 years, 15 females) and twenty-nine very old adults (85 {+/-} 3 years, 15 females) performed isometric dorsiflexion ramp contractions at 20% of maximal voluntary torque. Individual motor unit spike trains were identified from decomposed high-density surface electromyograms of the tibialis anterior. The paired motor unit technique was used to estimate the prolongation effect of PICs ({Delta}F and normalized {Delta}F), and the quasi-geometric approach to estimate neuromodulatory (brace height) and inhibitory pattern (attenuation slope) contributions. {Delta}F was lower in old and very old adults than in young adults, with no difference between old and very old adults regardless of sex. Compared with young adults, normalized {Delta}F was lower in both old and very old males, whereas in females it is only lower in the very old group. Brace height and attenuation slope did not differ across ages or sexes. These findings indicate that age-related changes in spinal motor neuron excitability are established by the seventh decade and do not progress further in healthy active very old adults, following a nonlinear trajectory that may differ between males and females. The absence of age-related differences in the neuromodulatory index does not support a lower monoaminergic drive in older adults.

Purpose: Impaired balance and fall risk in individuals with respiratory conditions highlight the importance of respiratory-postural integration during upright activity. However, breathing and postural control are often studied separately, limiting understanding of their interaction. Singing imposes concurrent ventilatory and trunk control demands, yet respiratory-postural responses during standing singing remain unclear. We examined the effects of vocalisation and tempo on respiratory-postural responses during standing, and whether age and singing experience moderated these responses. Methods: Eighty healthy younger (18-35yr; n=35) and older ([&ge;]60yr; n= 45) adults completed rhythmically paced speaking and singing tasks at slow, normal, and fast tempos while standing on a force platform. Respiratory rate (RR), mediolateral and anteroposterior sway, peripheral oxygen saturation, and perceived breathlessness were measured. Generalised estimating equations assessed vocalisation, tempo, and their interaction, with moderation by age and singing experience. Results: RR showed a vocalisation x tempo interaction (p<.001), with higher RR during slow speaking and greatest increases during fast singing. Singing increased postural sway relative to speaking, particularly mediolaterally (p<.001), with tempo-related increases in anteroposterior sway (p<.001). Older adults exhibited greater postural sway overall (p values[&le;].040) but similar task-dependent modulation to younger adults. Singing experience attenuated RR increases during fast singing (p=.033) and was associated with greater mediolateral sway (p=.004). Perceived breathlessness increased with tempo but was not associated with vocalisation; SpO2 was unchanged. Conclusions: Standing singing was associated with tempo-dependent respiratory and postural responses. Age and singing experience showed distinct associations with these responses. Task-dependent respiratory-postural coordination appeared preserved among participants.

PurposeLong COVID literature often describes new-onset symptoms or conditions following SARS-CoV-2 infection; however, Long COVID also encompasses the exacerbation of pre-existing conditions. We describe health conditions reportedly worsened by COVID-19 illness and associated concerns in U.S. adults. MethodsWe collected longitudinal data from adults 18 years and older participating in the Track Post COVID-19 Conditions, an active Long COVID cohort surveillance project. Respondents completed surveys 3-, 6-, and 12- months after COVID-19 illness, beginning in July 2023. In each survey, respondents were asked whether they had any health conditions or concerns that were made worse by COVID-19. Those with an affirmative response were asked to specify the worsening condition(s) via free text. Some respondents used this free text to describe challenges or experiences related to their worsening conditions. This report categorizes and quantifies reported conditions and concerns by body system categories and qualitatively describes associated challenges and experiences using direct quotes from respondents free text responses. ResultsAmong 2,227 adults, 263 (11.8%) respondents reported at least one worsening condition or concern following COVID-19 illness. Of these, nearly 32% reported a worsening respiratory condition or concern. Adults with worsening pre-existing conditions reported challenges accessing adequate care, completing daily activities, and managing their symptoms following COVID-19. DiscussionWhile Long COVID literature often focuses on the development of new-onset long-term symptoms, more than 1 in 10 adults reported worsening of a pre-existing condition following COVID-19. Reported challenges with accessing care and daily activities highlight potential need for accommodations or support for adults with Long COVID.

Reduced visibility is a common contributor to falls in older adults, but the neural processes that support safe locomotion under low lighting are not well established. In this study, 22 young (mean age = 22.5) and 20 older (mean age = 70.9) adults walked along a pathway under bright, ambient and dark lighting conditions while stepping over expected and unexpected obstacles projected on the floor. Mobile electroencephalography (EEG), combined with motion capture, was used to quantify spectral power changes during obstacle crossing preparation and the post-crossing phases, relative to a standing baseline. We identified that reduced lighting conditions elicited a widespread increase in cortical engagement, predominantly in the alpha band, during both preparation and reset phases. Obstacle negotiation recruited a distributed sensorimotor network, with reduced theta-alpha-beta desynchronisation during preparation and a robust post-movement beta rebound during the post-crossing reset phase. Expectation effects were modest, with unexpected obstacles inducing greater transient alpha-beta desynchronisation relative to expected obstacles, suggesting additional cortical resources are recruited under uncertainty. Age effects were minimal and limited to reduced frontal alpha desynchronisation in older adults compared to young adults during the preparation phase, possibly reflecting differences in walking speed rather than differences in neural control. In conclusion, environmental features such as low lighting and obstacle presence exert a stronger influence on cortical dynamics during walking than age and expectation effects alone. This provides a foundation for future work examining how these processes may differ in older adults at increased risk of falls.

Individuals with Down syndrome (DS) exhibit accelerated biological aging and unique patterns in disease prevalence. However, empirical characterizations of relative risk and age at onset of disease in this population are not common. Here we aimed to quantify the difference in risk and timing of age-related conditions between individuals with DS when compared to typically developing (TD) matched controls. We used semiparametric Cox proportional hazards models and Restricted Mean Event-Free Age (RMEA) to estimate differences in risk and approximate average age at onset of cataracts, cognitive decline, death, menopause, and osteoporosis. Individuals with DS were at higher risk of all age-related conditions or events and experienced these events at younger ages, excluding menopause. We observed a 24.5-fold higher risk of cognitive decline in individuals with DS (p<0.001), and between a 2- and 4-fold higher risk of the other age-related events or conditions (p<0.01). Cognitive decline and death occurred more than 14 years earlier in individuals with DS and were more likely to co-occur (p<0.001) compared to in TD individuals. These findings have important implications for tailored screening and prevention efforts in aging individuals with DS, as this population experiences a distinct cadence of age-related events.

Background: Mitochondrial dysfunction is a hallmark of ageing and is implicated in the pathogenesis of sarcopenia. Genetically confirmed mitochondrial disease provides a biologically definitive model of impaired oxidative phosphorylation against which novel functional assessments can be validated. We investigated whether a brief, submaximal, non-ischaemic intermittent isometric handgrip exercise protocol combined with Near Infra-Red Spectroscopy (NIRS) could detect distinct muscle oxygenation signatures in mitochondrial disease and sarcopenia. Methods: Adults with genetically confirmed mitochondrial disease (n=10, 7 female) and group-matched controls (n=19, 11 female), underwent assessment of maximal grip force, handgrip relaxation kinetics and a 3-minute intermittent submaximal isometric handgrip test (40% maximal voluntary contraction, 1Hz contraction/relaxation) with continuous force recording at 2000Hz and simultaneous NIRS. Findings were compared with 23 community-dwelling older adults from the Muscle Ageing and Sarcopenia Study (MASS) Lifecourse cohort (n=23, 13 female) including seven with probable or confirmed sarcopenia according to European Working Group on Sarcopenia in Older People (EWGSOP2) criteria. Results: Participants with mitochondrial disease demonstrated an attenuated reduction in tissue saturation index (TSI) during exercise compared with controls (TSI % {middle dot} s150-180s: -59.9 {+/-} 94.5 vs -175.1 {+/-} 128.4; p=0.02). Total haemoglobin (THb) responses did not differ between groups (p=0.10). In contrast, participants with probable or confirmed sarcopenia showed exaggerated reduction in TSI compared with non-sarcopenic participants (TSI % {middle dot} s150-180s: -256.5 {+/-} 146.0 vs -121.2 {+/-} 95.2; p=0.05), while THb responses remained similar. The association between sarcopenia and TSI differed by sex (interaction coefficient -240, p=0.016), with greater TSI reductions observed in males with sarcopenia. Differences in TSI between mitochondrial disease and control groups were independent of age and sex. No statistically significant associations were observed between THb responses and age, sex or force production. Conclusions: A brief, non-ischaemic handgrip exercise protocol combined with NIRS identified divergent muscle oxygenation responses in mitochondrial disease and sarcopenia. The attenuated TSI response observed in genetically confirmed mitochondrial disease is consistent with impaired oxygen utilisation secondary to defective oxidative phosphorylation, whereas the exaggerated TSI reduction observed in sarcopenia suggests a distinct physiological mechanism, potentially involving impaired oxygen delivery, reduced muscle efficiency, or both. This portable and non-invasive approach has significant potential as a physiological biomarker for mechanistic studies and clinical trials in ageing and neuromuscular disease. More broadly, our findings demonstrate how biologically defined rare diseases can provide a powerful framework for interpreting mechanisms in common heterogeneous age-related conditions.

Importance Several influenza vaccine products are available for older adults, but evidence combining both their absolute effectiveness and head-to-head comparisons within the same large population remains scarce, limiting product-specific vaccination guidance for this high-risk group. Objective To estimate the relative and absolute effectiveness of influenza vaccines against influenza-related hospitalization and in-hospital mortality among older adults. Design, Setting, and Participants Nationwide cohort study using the French National Health Data System (SNDS), including all adults born in 1958 or earlier, living in metropolitan France, at the start of the 2023-2024 campaign. Relative effectiveness (rVE) was assessed using a landmark design with exposure fixed before epidemic onset. Absolute effectiveness (VE) was assessed using a sequential trial emulation with weekly matching of newly vaccinated individuals to yet unvaccinated controls. Follow-up ran from December 5, 2023 to April 19, 2024. Exposures Vaccination with a high-dose (Efluelda) or one of three standard-dose quadrivalent vaccines (Influvac, Fluarix, Vaxigrip), compared individually or by dosage category, and against no vaccination. Main Outcomes and Measures Hospitalization with a diagnosis of influenza and death during such hospitalization. Effectiveness was estimated as 1 minus the hazard ratio from weighted Cox models, with corresponding numbers needed to vaccinate to prevent one case. Results Among 13.8 million eligible individuals, 5.7 million (41%) were vaccinated at the beginning of follow-up. The high-dose vaccine showed greater rVE than standard-dose vaccines against hospitalization (27% [95%CI, 23 to 31%]) and in-hospital death (27% [95%CI,14 to 38%]). Absolute VE was 39% (95%CI, 38 to 41%) against hospitalization and 45% (95%CI, 40 to 49%) against in-hospital death, and was higher for the high-dose vaccine (50%) than standard-dose vaccines (36%) against hospitalization. This difference was most evident among adults 70 years and older, while overall effectiveness declined with age. Conclusions and Relevance In this study, influenza vaccination conferred moderate protection against severe outcomes in older adults, and high-dose vaccination was associated with significantly greater effectiveness than standard-dose vaccines, particularly among adults 70 years and older. These findings support product-level differentiation in vaccination strategy for this population.

Consumer-grade wearables can continuously and longitudinally measure health signals such as heart rate (HR), activity, and sleep. While changes in these signals have been implicated in the early detection of infection (e.g., influenza, SARS-CoV-2) and real-time alerting systems have been developed, evidence from prospective deployment in pediatric transplant households remains limited. Here, we study the application of a resting heart rate (RHR)-based real-time alerting algorithm to detect respiratory virus infection in a cohort of solid organ transplant recipients and their household members (n=146) in a Phase IV clinical trial (NCT06161454). Reliable and efficient alerting for infection events may be particularly relevant for this population, enabling isolation, clinical management, and/or treatment. Of 31 antigen-test outcomes, nearly all (29/31, 93.5%) correspond to either influenza, COVID, or other patient-reported symptoms. We describe RHR, HR, activity, and sleep/circadian rhythm features around clinical alarm events, identifying candidate features for further investigation and algorithm improvement.

The ovary ages earlier than most other organs, but the molecular mechanisms that drive this decline remain unclear. To gain more insight into this process, we profiled ovaries from young and reproductively aged mice after synchronized ovulation and found that aging was marked by the accumulation of luteal cells with increased steroidogenic, antigen presentation and inflammatory gene expression. DNA methylation was globally preserved in aged ovaries but showed focal age-associated changes at gene regulatory regions. Protein profiling identified an increase in checkpoint kinase 2 (CHK2), a DNA damage response protein encoded by the CHEK2 gene associated with the timing of natural menopause in humans. CHK2 and DNA damage markers were increased in aged mice and human ovarian somatic cells. Transient CHK2 inhibition in aged mice increased the proportion of mature oocytes and blastocyst formation and reversed inflammatory signatures in granulosa cells. These findings identify defective luteal resolution and inflammation as features of ovarian aging and suggest that CHK2 induces a reversible age-associated inflammatory signature. Our results show that reproductive decline may remain pharmacologically modifiable during ovarian aging via regulation of the inflammatory response.