Longevity Papers

Elucidating the role of somatic mutations in cancer, healthy tissues, and aging depends on methods that can accurately characterize somatic mosaicism across different cell types, as well as assay their impact on cellular function. Current technologies to study cell-type-specific somatic mutations within tissues are low-throughput. We developed Duplex-Multiome, incorporating duplex consensus sequencing to accurately identify somatic single-nucleotide variants (sSNV) from the same nucleus simultaneously analyzed for single-nucleus ATAC-seq (snATAC-seq) and RNA-seq (snRNA-seq). By introducing strand-tagging into the construction of snATAC-seq libraries, duplex sequencing reduces sequencing error by >10,000-fold while eliminating artifactual mutational signatures. When applied to 98%/2% mixed cell lines, Duplex-Multiome identified sSNVs present in 2% of cells with 92% precision and accurately captured known sSNV mutational spectra, while revealing unexpected subclonal lineages. Duplex-Multiome of > 51,400 nuclei from postmortem brain tissue captured sSNV burdens and spectra across all major brain cell types and subtypes, including those difficult to assay by single-cell whole-genome sequencing (scWGS). This revealed for the first time that diverse neuronal and glial cell types show distinct rates and patterns of age-related mutation, while also directly discovering developmental cell lineage relationships. Duplex-Multiome identified clonal sSNVs occurring at increased rates in glia of certain aged brains, as well as clonal sSNVs that correlated with changes in expression of nearby genes, in both neurotypical and autism spectrum disorder (ASD) individuals, directly demonstrating that somatic mutagenesis can contribute to gene expression phenotypes. Duplex-Multiome can be easily adopted into the 10X Multiome protocol and will bridge somatic mosaicism to a wide range of phenotypic readouts across cell types and tissues.

Single-cell RNA sequencing (scRNA-seq) has significantly advanced our understanding of retinal aging, yet the specific molecular characteristics within cell populations remain incompletely defined. We profiled 223,612 single cells from 18 unfrozen human retinas obtained from 13 Chinese donors aged 34-92 years, providing an ethnically specific atlas across the adult lifespan. A sparsity-driven machine-learning (ML) pipeline (L1-regularized logistic regression, recursive feature elimination with cross-validation) identified age-discriminatory genes within each major retinal cell type, complemented by gene set scoring for cellular senescence and metabolic pathways. Using integrated differential expression and ML feature selection, we identified eleven major retinal cell populations and observed aging-associated shifts. ML classifiers achieved high accuracy (80-96%), particularly for microglia (96%), revealing mitochondria-centric aging signatures in rods and bipolar cells, proteostasis and retinoid metabolism in cones, and structural-RNA maintenance signatures in horizontal cells. This study delivers the first ML-derived, cell-type-specific aging gene signatures for the human retina in a Chinese cohort, offering a reference for population-tailored biomarker discovery.

In the elderly population, age-related macular degeneration (AMD) is a major cause of visual impairment, characterized by a thinner retinal pigment epithelium and loss of photoreceptors. 6-shogaol (6S), a component of dried Zingiber officinale Roscoe, has been studied for its multiple therapeutic effects. The current study aimed to investigate the effect of 6S supplementation on AMD. 25-month-old C57BL/6 mice were orally administered with 10 mg/kg of 6S for 28 consecutive days. The thickness of the retinal layer was measured by histological analysis. mRNA expression related to fibrosis, inflammation and endoplasmic reticulum stress was measured by real-time polymerase chain reaction. As a result, 6S increased the thickness of the retinal layer and promoted postsynaptic density protein-95 expression in the outer plexiform layer of the aged mice. Moreover, 6S suppressed ocular mRNA expression related to the fibrotic process, including transforming growth factor beta, collagen type 1 alpha 1, and alpha smooth muscle actin. Furthermore, 6S reduced pro-inflammatory cytokines including tumor necrosis factor alpha, interleukin 1 beta, cyclooxygenase-2, and inducible nitric oxide synthase in the eyeballs of aged mice. Lastly, 6S inhibited ocular endoplasmic reticulum stress measured by mRNA expression of C/EBP homologous protein and spliced X-box binding protein-1 in the aged mice. Taken together, these findings suggest that 6S and dried ginger could be a potential nutraceutical candidate for AMD or other age-related eye diseases.

Hair graying and loss are significant indicators of aging, and oxidative stress plays an essential role in this progression. Ergothioneine (EGT), potent antioxidant from edible fungi and certain prokaryotes, regulates the antioxidant defense system. However, its anti-aging effects on hair follicles (HFs) remain obscure. We used dermal papilla cells (DPCs) co-culture system, HFs organ culture system, aged mice and H

Investigations into the association between serum 25-hydroxyvitamin D (25(OH)D) levels and the risk of age-related ocular diseases have yielded inconsistent results. Thus, we aimed to provide robust longitudinal evidence, identify optimal serum thresholds, and explore the underlying mechanisms.

Decline of skeletal muscle function in old age is a significant contributor to reduced quality of life, risk of injury, comorbidity and disability and even mortality. While this loss of muscle function has traditionally been attributed to sarcopenia (loss of muscle mass), it is now generally appreciated that factors other than mass play a significant role in age-related muscle weakness. One such factor gaining increased attention is the ectopic accumulation of lipids in skeletal muscle, in particular, intramyocellular lipids (IMCLs). It has been appreciated for some time that metabolic flexibility of several tissues/organs declines with age and may be related to accumulation of IMCLs in a "vicious cycle" whereby blunted metabolic flexibility promotes accumulation of IMCLs, which leases to lipotoxicity, which can then further impair metabolic flexibility. The standard interventions for addressing lipid accumulation and muscle weakness remain diet (caloric restriction) and exercise. However, long-term compliance with both interventions in older adults is low, and in the case of caloric restriction, may be inappropriate for many older adults. Accordingly, it is important, from a public health standpoint, to pursue potential pharmacological strategies for improving muscle function. Because of the success of incretin-analog drugs in addressing obesity, these medications may potentially reduce IMCLs in aging muscles and thus improve metabolic flexibility and improve muscle health. A contrasting potential pharmacological strategy for addressing these issues might be to enhance energy provision to stimulate metabolism by increasing NAD + availability, which is known to decline with age and has been linked to reduced metabolic flexibility. In this narrative review, we present information related to IMCL accumulation and metabolic flexibility in old age and how the two major lifestyle interventions, caloric restriction and exercise, can affect these factors. Finally, we discuss the potential benefits and risks of select pharmacologic interventions in older adults.

Mechanistic target of rapamycin complex 1 (mTORC1) senses nutrient availability to orchestrate metabolic processes critical for physiological homeostasis and organismal ageing. While mTORC1 preferentially regulates the translation of 5\'-terminal oligopyrimidine (TOP) motif-containing mRNAs that predominantly encode ribosomal proteins (RPs) via the translational repressor 4E-BP, this mTORC1 function is resistant to rapamycin inhibition. TOP mRNAs are exceptionally abundant, thus imposing a major translational burden on cells; yet how their translation is physiologically tuned and linked to longevity remain unexplored. Here we identify Lsp2, originally known as a storage protein, as an adipose effector of mTORC1 that modulates lifespan in Drosophila. Lsp2 is induced by essential amino acids (EAAs) via mTORC1. Genetic ablation of Lsp2 to blunt organismal response to protein diets drives robust lifespan extension without compromising key life-history traits such as reproduction. Translatomic profiling reveals that loss of Lsp2 selectively reduces global TOP mRNA translation in a 4E-BP-dependent manner, thereby extending lifespan via a mechanism distinct from rapamycin inhibition that fails to reduce 4E-BP phosphorylation in vivo. Finally, Lsp2 adipokine promotes 4E-BP phosphorylation and acts systemically across tissues to shape the lifespan responses to dietary protein. Collectively, our findings establish Lsp2 as a novel translational regulator of TOP genes that mechanistically couples physiological ribosomal protein synthesis with organismal longevity.

Taurine, the most abundant sulfonic amino acid in humans is largely obtained from diets rich in animal proteins. However, taurine is dietary non-essential because it can be synthesized from cysteine by activation of transsulfuration pathway (TSP) when food consumption is low or if the diet is predominantly plant based. The decline of taurine was proposed as the driver of aging through an undefined mechanism. Here, we found that mild food restriction in humans for one year that resulted in 14% reduction of calorie intake elevated the hypotaurine and taurine concentration in adipose tissue. Therefore, we investigated whether elevated taurine mimics beneficial effects of caloric restriction on inflammation, a key mechanism of aging. Interestingly, aging increased the circulating and tissue concentrations of taurine suggesting that elevated taurine may serve as a hormetic stress response metabolite that regulates mechanism of age-related inflammation. The elevated taurine protected mice against mortality from sepsis and inhibited inflammasome-driven inflammation and gasdermin-D (GSDMD) mediated pyroptosis. Mechanistically, danger signals including hypotonicity that activate NLRP3-inflammasome, caused upstream taurine efflux from macrophages, which triggered potassium (K+) release and downstream canonical NLRP3 inflammasome assembly, caspase-1 activation, GSDMD cleavage and IL-1{beta} and IL-18 secretion that was reversed by taurine restoration. Notably, taurine does not efflux from GSDMD pore and inhibited IL-1{beta} from macrophages independently of known transporters SLC6A6 and SLC36A1. Increased taurine in old mice promotes healthspan by inducing anti-inflammatory pathways previously linked to youthfulness. These findings demonstrate that taurine is an upstream metabolic sensor of cellular perturbations that control NLRP3 inflammasome and lowers age-related inflammation.

An attempt was made to rejuvenate cumulus cells (CCs) of women of advanced maternal age (AMA) using extracellular vesicles (EV) isolated from the follicular fluid (FF) of young donors. FF samples were taken from healthy women aged 23-26 during the ART program. CCs from patients of advanced maternal age (36-47 years) were co-cultured with EV of young donors. The target genes were selected based on their functions in CC and early embryogenesis (PFKP, EREG, UBE2T, HAS2, VCAN, CYP19A, GREM1, STS, BAX, PTGS2, BCL2, SPSB2, AREG, CCNB1, EGFR). Comparative analysis of all women without stratification by the type of final follicle maturation drug (hCG or gonadotropin-releasing hormone agonist, GnRHa) revealed no statistically significant difference in the expression of the selected genes. In the group treated with GnRHa, a statistically significant increase in the expression of HAS2 (by 1.8 times, p = 0.04), GREM1 (by 1.4 times, p = 0.08), and BAX (by 1.4 times, p = 0.04) in CCs was observed after their co-culturing with FF EVs from young donors. The findings suggest the potential for the "rejuvenation" of CC in women of advanced maternal age when aGnRH is used in combination with FF EVs from young women, because the expression of apoptosis genes decreases and the expression cell proliferation gene increases during co-culturing of CC with FF EVs.

Idiopathic pulmonary fibrosis (IPF) is a prototype of chronic, progressive, and fibrotic lung disease. While advancing age is recognized as the most significant risk factor for both the development and mortality associated with pulmonary fibrosis, precise mechanisms underlying this association remain elusive. Here, Nephronectin (NPNT) is identified as an antiaging molecule, a potential major regulator of the progression of pulmonary fibrosis. In IPF patients, a marked reduction in NPNT expression is detected in lung tissues, which correlated with a decline in lung function. The study reveals that NPNT deficiency exacerbates bleomycin-induced senescence in alveolar epithelial cells, potentially intensifying fibrosis severity due to diminishes extracellular matrix turnover. Conversely, NPNT overexpression in the alveolar epithelium improves lung respiratory function and enhances resistance to aging and fibrosis. Mechanistically, NPNT inhibits the hyperactivation of LATS1 and MOB1, facilitates YAP1 nuclear translocation, and suppresses YAP1 ubiquitination and degradation, contingent upon the interaction between NPNT and ITGA3. Notably, pharmacological elevation of NPNT protein levels using Escin has been shown to alleviate pulmonary fibrosis and improve lung function in mice. The findings shed light on the key mechanism underlying stress-induced senescence and fibrosis, and offer a promising framework for interventions targeting aging-related diseases.

The gut microbiota evolves over a lifetime and significantly impacts the aging process. Targeting the gut microbiota represents a novel avenue to delay aging and aging-related physical and mental decline. However, the underlying mechanism by which the microbiota modulates the aging process, particularly age-related physical and behavioral changes is not completely understood. We conducted fecal microbiota transplantation (FMT) from young or old male donor mice to the old male recipients. Old recipients with young microbiota had a higher alpha diversity than the old recipients with old microbiota. Compared to FMT with old microbiota, FMT with young microbiota reduced body weight and prevented fat accumulation in the old recipients. FMT with young microbiota also lowered frailty, increased grip strength, and alleviated depression and anxiety-like behavior in the old recipients. Consistent with observed physical changes, untargeted metabolomic analysis of serum and stools revealed that FMT with young microbiota lowered age-related long-chain fatty acid levels and increased amino acid levels in the old recipients. Bulk RNAseq analysis of the amygdala of the brain showed that FMT with young microbiota downregulated inflammatory pathways and upregulated oxidative phosphorylation in the old recipients. Our results demonstrate that FMT with young microbiota has substantial positive influences on age-related body composition, frailty, and psychological behaviors. These effects are associated with changes in host lipid and amino acid metabolism in the periphery and transcriptional regulation of neuroinflammation and energy utilization in the brain.

Despite the manifestation and contribution of cellular senescence to aging and various diseases, accurate identification of heterogeneous senescent cells remains challenging. Current senescence evaluation methods rely mainly on limited markers or homogeneous samples, which might fail to capture universal senescence features, limiting their generalizability. Here we developed the human universal senescence index (hUSI), an accurate and robust senescence evaluation method for diverse cells and samples. Based on features learned from the most comprehensive cellular senescence-associated transcriptome data so far, hUSI demonstrated its convincing connections with senescence phenotypes and superior robustness in predicting senescence state. Using hUSI, we discovered potential senescence regulators and mapped senescent cell accumulation across cell types in coronavirus disease 2019 (COVID-19). The method also facilitates decoding heterogeneous senescence states in melanoma tumors, identifying prognosis-associated signaling pathways. Overall, hUSI demonstrates its utility in characterizing cellular senescence across biological contexts, with broad applications in aging research and clinical practice.

The decline in neutrophil antimicrobial function renders vaccinated aged hosts less protected against Streptococcus pneumoniae infection. In vaccinated hosts, activation of neutrophils via phagocytic receptors, like complement and Fc{gamma} receptors, mediates bacterial uptake and killing. However, the mechanisms behind changes in signaling downstream of these receptors with age is not known. Mitogen-activated protein kinases (MAPK) are signaling cascades activated in response to cell surface receptors. Using bone marrow neutrophils isolated from young and aged mice, and MAPK phosphorylation array, we found differences in MAPK activation in an opsonin dependent manner (complement vs antibody). Neutrophils from aged mice failed to increase MAPK phosphorylation upon infection with pneumococci opsonized with heat-inactivated (HI) sera from hosts immunized with the pneumococcal conjugate vaccine (PCV), indicating an age-related defect in Fc{gamma}R signaling. Neutrophils from aged mice had higher basal levels of MAPK phosphorylation when compared to young controls, including a 15-fold increase in ERK1/2 phosphorylation. Inhibition of ERK1/2 signaling blunted pneumococcal killing by PMNs from young mice, but improved killing by neutrophils from aged mice, only when the bacteria were opsonized with HI immune but not naive sera. In young human participants, in vitro inhibition of ERK1/2 in neutrophils resulted in decreased pneumococcal killing, but only following PCV vaccination, suggesting that the effects of ERK1/2 inhibition are clinically relevant in vaccinated hosts. This study identifies age-related changes in ERK1/2 activation and demonstrates that balanced activation of this pathway is crucial for neutrophil antimicrobial activity against antibody opsonized bacteria following vaccination in both mice and humans.

Animals store oocytes in a dormant state for weeks to decades before ovulation. The homeostatic programs that oocytes use to endure long-term storage are poorly understood. Using female nematodes as a short-lived model, we found that oocyte formation and storage required IFET-1, the conserved eIF4E-transporter protein (eIF4ET). IFET-1 co-assembled with CAR-1 (Lsm14) to form micron-scale condensates in stored oocytes, which dissipated after oocyte activation. Depletion of IFET-1 destabilized the stored oocyte proteome, leading to lower translation, a decline in microtubule maintenance proteins, and errors in microtubule organization and meiotic spindle assembly. Deleting domains within IFET-1 impaired oocyte storage without affecting oocyte formation. Thus, in addition to establishing a healthy oocyte reserve in young mothers, IFET-1 ensures that correct levels of cytoskeletal proteins are maintained as oocytes age. eIF4ET also localized to micron-scale puncta in dormant human oocytes. Our results clarify how eIF4ET maintains the oocyte reserve and further support eIF4ET dysfunction as an upstream cause of embryonic aneuploidy and age-related infertility.

The intersection of aging and reproductive decline presents a significant challenge in human health, with fertility rates decreasing sharply in later life for both sexes. This review delves into the intricate relationship between germ cells, the fundamental units of reproduction, and their surrounding microenvironment, known as the niche. Emphasizing that reproductive longevity is not solely determined by the intrinsic properties of germ cells, but rather by the complex interplay with their niche, a dynamic system that changes with age. We highlight evidence from model organisms like Drosophila and C. elegans demonstrating how age-related changes in niche signaling impact germ cell function. A systems biology approach, integrating multi-omics data (genetics, epigenetics, cellular behavior), is crucial to fully understanding this complex interaction. Specifically, we discuss the role of epigenetic modifications, such as DNA methylation and histone acetylation, in modulating niche-germ cell communication. This approach offers a comprehensive view of the aging reproductive system and opens up avenues for therapeutic interventions aimed at modulating the niche and potentially extending reproductive lifespan. Future research focused on unraveling the specific molecular mechanisms underlying the niche-germ cell interaction will be pivotal in developing strategies to combat age-related reproductive decline.

Glycation is a non-enzymatic post-translational modification associated with aging and disease. Because it occurs spontaneously, it is extremely difficult to control the extent of glycation at distinct sites within target proteins, especially in cellular systems. Here we report a chemical approach, referred to as \"dialAGE\", that enables the site-specific control of protein glycation. This unique tool requires the introduction of just a single point mutation that modulates the glycation susceptibility of a nearby arginine. As proof-of-concept, extensive mass spectrometry analysis was performed to confirm that dialAGE can modulate site-specific glycation levels at multiple arginine residues in ubiquitin in vitro, enabling both enhanced and diminished glycation. Introduction of dialAGE point mutations and/or glycation with the biologically relevant glycating agent methylglyoxal did not affect polyubiquitin chain formation using in vitro ubiquitination assays. Furthermore, we show that dialAGE can be used to modulate ubiquitin glycation levels in living mammalian cells. We therefore anticipate that this method will be particularly useful for enabling the study of glycation as a genuine, functional, post-translational modification.

The axolotl (Ambystoma mexicanum) has a great capacity to regenerate its tissues; however, the fidelity and success of its regenerative process diminish with age. Retrotransposons make up the largest portion of the axolotl genome, and their expression may be involved in this age-related decline. Through an integrative analysis of repetitive element expression using RNA sequencing, it is shown that Ty3 retrotransposons are highly upregulated in the axolotl as an effect of chronological aging. Other non-long-terminal-repeat transposons, including long interspersed nuclear element 1, function as hubs of gene coexpression networks involved in muscle development and regulation of apoptosis and connective tissue replacement, which are also suppressed in adulthood. By contrast, it is found that during regeneration of the limb, these pathways and the expression of Ty3 retrotransposons are distinctly downregulated. Although the blastema can readjust most of the transposon differential expression in adulthood, several elements remain affected and may have an impact in the immune response during regeneration. This analysis provides a profile of retrotransposon expression through chronological aging and during limb regeneration in the axolotl and indicates that transposons are responsive to physiological changes in a tissue-specific way and may participate in the gene coregulatory networks underlying the regenerative process.

Targeting partial cellular reprogramming pathways through specific small molecule combinations holds promise for lifespan extension in model organisms. Chemical cocktails like RepSox and tranylcypromine (TCP) may induce beneficial age-related changes without the risks of full reprogramming. This study investigated the effects of RepSox and TCP on neurological markers, physical activity, skeletal health, and survival in aging C3H female mice.

Senescence is a pivotal cellular process, which also plays a major role in development, immune regulation, tissue repair, and aging, triggered by stressors such as telomere shortening, oncogene activation, and DNA damage. Characterized by distinct morphological and molecular features, senescence is known to act as a tumor suppressive mechanism through irreversible cell cycle arrest. However, emerging studies reveal a paradox: prolonged senescence in cancer cells can drive tumorigenesis via the senescence-associated secretory phenotype, promoting proliferation, invasion, and metastasis. This comprehensive review elucidates the molecular intricacies of senescence to induce growth arrest, enhance immune surveillance, and favorably modulate the tumor microenvironment to inhibit cancer progression. Additionally, it examines the senescence-inducing effects of conventional therapies and explores the potential of emerging therapies, including targeted therapies and chimeric antigen receptor T cell therapy. The present review also highlights the promise of senotherapeutic strategies in selectively targeting senescent cells to improve therapeutic outcomes. It discusses the innovative integration of machine learning tools for biomarker discovery and patient stratification offering a transformative approach to improve cancer treatment efficacy.

Sarcopenia is defined as a progressive decrease in skeletal muscle mass and strength due to ageing, and is associated with adverse outcomes such as falls, fractures, and physical disability, leading to increased healthcare expenditures and mortality risks. Although sarcopenia has recently become a major challenge, there are currently no effective interventions or drugs marketed for sarcopenia. Therefore, new biomarkers for early diagnosis and targeted drugs for the treatment of sarcopenia are urgently needed. In the present research, transcriptomics and proteomics analyses were combined and experiments at the transcriptional and protein levels were conducted to identify key genes and molecular mechanisms underlying sarcopenia in senescence-accelerated mouse prone 8 (SAM-P8) mice, followed by molecular docking to predict targeted drugs. The combined omics analysis identified 8 key genes, while the experiments verified that only CD9 significantly decreased in sarcopenia. The gene set enrichment analysis (GSEA) results suggested that CD9 involved in ATP biosynthesis, mitochondrial biogenesis, and oxidative phosphorylation. Besides, dapoxetine, levomilnacipran, and milnacipran were predicted to target CD9 through molecular docking. Our study reported for the first time that CD9 is a novel potential biomarker of sarcopenia, and targeting CD9 may be a new idea for the development of therapeutic drugs for sarcopenia.

Productive lifespan is a critical economic trait for both dual-purpose and dairy cows, as it determines lifetime milk production. Xinjiang Brown cattle, a dual-purpose breed widely raised in China's Xinjiang region, have a population of nearly two million and play a vital role in the local economy. However, the molecular mechanisms influencing aging and productive lifespan in Xinjiang Brown cattle remain largely unknown. In this study, we collected white blood cell (leukocyte) transcriptome data from 66 Xinjiang Brown cattle, aged 31 to 160 months, to investigate the dynamic changes in their gene expression profiles across different ages and identify genes potentially influencing their aging process.

Dietary strategies play an important role in determining longevity and healthy aging. There is an increasing awareness of dietary factors and sarcopenia, which presents as a decline in muscle mass and function, mainly in the senior population. The aim of this study is to explore the association of adherence to the Dietary Guideline (DG) on muscle health in the elder Taiwanese. We included 410 adults aged 65 or older, of whom 189 (46.1%) were women, in the Nutrition and Health Survey in Taiwan (NAHSIT) during 2014-2016 with comprehensive records of food frequency questionnaires and 24-h dietary recalls, as well as measurements of dual-energy X-ray absorptiometry (DXA) for muscle mass and 8-m gait speed for physical performance. A novel indicator, named Healthy Eating Index for Taiwanese (HEI-TW), was developed as an amended version of HEI-2015 to assess how dietary patterns adhere to the DG of Taiwan. Participants who fulfilled "low muscle mass" and "poor physical performance" defined by the Asian Working Group for Sarcopenia (AWGS) criteria were recognized as having sarcopenia; others, in contrast, were recognized as healthy controls. Seventy-one (17.3%) participants categorized into the sarcopenia group had significantly lower mean scores of overall HEI-TW, total vegetables, unrefined grains, and dairy products. The adjusted odds ratios (95% confidence intervals) for sarcopenia were 0.98 (0.95-1.00) for overall HEI-TW, 0.78 (0.66-0.94) for total vegetables, and 0.85 (0.76-0.95) for dairy, respectively, for one unit increase in these components. In subgroup analysis stratified by sex and age, the overall HEI-TW was inversely associated with the odds of sarcopenia among men and individuals aged 75 or older. This study showed that a higher HEI-TW score adhering to Taiwan DG and abundant consumption of total vegetables and dairy products is associated with lower prevalence of muscle loss and sarcopenia in elder Taiwanese.

Most fungi can produce dormant, long-lived cells known as spores. Spores play a critical role in fungal biology and human health, but much about spores is unknown. Here, we investigate factors affecting spore fitness using the fission yeast Schizosaccharomyces pombe as a model. We found that storage conditions affect spore longevity, and that spore health declines over time. We identified a delay in dormancy breaking (germination), decreased asymmetry during cell division, and reduced stress tolerance as aging phenotypes. These results support that S. pombe spores are affected by both time and experiences during dormancy, highlighting critical features of spore biology and revealing parallels between aging in spores and aging in animal cells.

Sleep is a biological necessity and fundamental to health. However, the associations of sleep patterns (integrating sleep determinants) with DNA methylation age acceleration (DNAm AA) remain unknown. We aimed to investigate the associations of sleep patterns with DNAm AA.

The regulatory ability of bone marrow stem cells (BMSC) to chemokines and inflammatory factors has a significant effect in a variety of diseases. It is very important to delay the aging of BMSC and restore the function of aging BMSC.

Excess cellular senescence contributes to age-related increases in frailty and reductions in skeletal muscle strength. In the present study, we determined the efficacy of oral intermittent treatment (1 week on-2 weeks off-1 week on) with the natural flavonoid senolytic fisetin to improve frailty and grip strength in old mice. Further, the effects of fisetin on physical function were evaluated in young mice. We performed bulk RNA sequencing of quadricep skeletal muscle to determine the cell senescence-related signaling pathways modulated by fisetin. We also assessed the relative effects of fisetin on frailty and grip strength with aging in comparison with two other well-established approaches for the removal of senescent cells: (1) genetic-based clearance of excess senescent cells in old p16-3MR mice, a model that allows for clearance of p16-positive (p16+) senescent cells, and (2) oral intermittent treatment with the synthetic pharmacological senolytic ABT-263 in old mice. We found that fisetin mitigated the adverse changes in frailty and grip strength with aging. Fisetin had no effects in young mice. The improvements in frailty and grip strength in old mice were accompanied by favorable modulation of the skeletal muscle transcriptome, including lower abundance of cellular senescence-related genes (e.g., Cdkn1a and Ddit4). Improvements in frailty and grip strength with fisetin were comparable to those observed with genetic-based clearance of excess p16+ senescent cells and treatment with ABT-263. Taken together, our findings provide proof-of-concept support for fisetin as a senolytic strategy to improve physical function with aging.

Suppression of the insulin-IGF-mTORC1-Ras network ameliorates aging in animals. Many drugs have targets in the network because of its roles in cancer and metabolic disease and are candidates for repurposing as geroprotectors. Rapamycin, an established geroprotective drug, blocks mTORC1 signaling, and trametinib inhibits the Ras-MEK-ERK pathway. In this study, we assessed survival and health of male and female mice treated with trametinib, rapamycin or their combination. We show here that trametinib treatment extended lifespan in both sexes and that its combination with rapamycin was additive. Combination treatment reduced liver tumors in both sexes and spleen tumors in male mice, blocked the age-related increase in brain glucose uptake and strongly reduced inflammation in brain, kidney, spleen and muscle and circulating levels of pro-inflammatory cytokines. We conclude that trametinib is a geroprotector in mice and that its combination with rapamycin is more effective than either drug alone, making the combination a candidate for repurposing as a gerotherapy in humans.

The rising global demographic aging and the subsequent increase in the prevalence of age-related diseases highlight the need to understand aging biology. A key player in organismal aging is the immune system, which has broad systemic effects. On the one hand, immune aging involves the decline of hematopoietic stem cells and significant alterations in the functionality and composition of both innate and adaptive immunity. On the other hand, the aged immune system contributes to chronic inflammation and disrupted tissue homeostasis, thereby driving systemic aging processes. In this review, we examine the close interaction between aging and the immune system and discuss emerging therapeutic strategies aimed at modulating immune function to mitigate age-related pathologies.

Cellular senescence represents a stable cell cycle arrest state that plays critical roles in tissue aging and age-related pathologies. While single-cell RNA sequencing (scRNA-seq) has enabled comprehensive profiling of millions of cells in aged tissues, reliable identification of senescent cells remains challenging due to the weak and non-specific expression of marker genes. Although existing computational methods could be used to detect active cell states through gene set scoring approaches, their performance with senescence genes has not been rigorously assessed. In this study, we showed that senescence genes display weak, non-specific expression patterns across different tissues and are susceptible to dropout events in scRNA-seq. Current scoring approaches suffered intrinsic limitations in single-cell detection using weakly expressed markers. Simply expanding the gene set of weak markers failed to improve detection accuracy. To overcome these limitations, we developed ICE (Iterative-imputation-based Cell Enrichment), a computational framework that combines expression imputation with iterative marker refinement. ICE achieved a remarkable improvement in detection precision when analyzing pancreatic cells with weakly expressed markers, by increasing accuracy from 56% to 98%. Applied to senescence markers (CDKN1A/p21, CDKN2A/p16, ATF3, and MX1), ICE successfully identified marker-specific cell populations, including stressed {beta} cells in aging and type-I interferon-responsive microglia in Alzheimer\'s disease (AD). Together, our study offered a rigorous analytical framework for single cell detection using markers with weak expressions, which will facilitate in-depth exploration of senescence heterogeneity and temporal dynamics in human tissue and disease states.

Necrosis is uncontrolled cell death that marks the irreversible threshold of biological degeneration. Rooted in the Greek nekros (death), it is a pivotal mechanism underlying numerous diseases, including cancer, as well as renal, cardiac, neuronal, and hepatic disorders, and more broadly, the aging process. Despite its profound impact on morbidity and mortality, necrosis remains untreatable and has long been viewed as a chaotic, unavoidable aspect of biology. This review examines the mechanisms of necrosis and outlines its far-reaching impact on health, as revealed by emerging evidence. Furthermore, we explore its potential as a game-changing therapeutic target. Inhibiting necrosis could revolutionize treatments for acute and chronic age-related conditions like cancer, kidney disease, cardiovascular disease (including heart attacks and strokes), and neurodegeneration, while also preserving resilience-and even slowing aging itself. Beyond Earth, where microgravity, cosmic radiation, and oxidative stress accelerate cellular decline, targeting necrosis may also hold the key to preserving astronaut resilience and health on long-duration space missions, offering insights that could reshape human longevity both on and off the planet.

Degeneration of the auditory and vestibular hair cells (HCs) leads to dysfunction of two essential senses: hearing loss and decompensated balance perception. However, the cellular and molecular mechanisms governing inner ear aging and its link to dysfunctions of sensory HCs remain unclear. Here, we constructed an aging-associated cell atlas of cochlear and utricular tissues in C57BL/6J mice through single-nucleus RNA sequencing, revealing transcriptionally distinct hair cell subtypes and spatially restricted molecular markers that delineate specific cellular populations. We uncovered activation of macrophages and shaped inflamed niches in the aged inner ear. Furthermore, we demonstrated the cell type-specific signatures of aging between the two organs, with focused characterization in six HC subtypes uncovering core mechanisms like dysregulated RNA splicing underlying degeneration. Crucially, suppressing RNA splicing factor Rbm25 abolished inflammation-induced HC deterioration. Our study reveals complex, multifactorial mechanisms that underlie inner ear aging and offer potential targets for preventing HC degeneration.

Senescence causes deterioration in the functioning and physiology of an organism. Microglia, the standing resident immune brain cells transform from neuroprotective to neurotoxic with age. Rapid process motility and cellular migration of microglia in the developing brain, and other characteristics are regarded to be crucial for immunological defense and tissue repair. As they mature, microglia not only differ in their morphology but also in their functioning. However, the exact mechanism related to the atrophies caused by aged microglia or their role in neurodegenerative diseases is still uncertain. The aim of this updated review is to provide insights of how aging microglial cells change and how this influences the development of neurodegenerative diseases. As life expectancy rises, there is an increase in the accumulation of iron, ROS/NOS, protein misfolding and insufficient clearing of debris. This is attributed to the age-dependent alterations in the genes linked to energy metabolism, mitochondrial and lysosome function, and neuroinflammation. Aging microglia often shifts towards a pro-inflammatory state with a reduction of anti-inflammatory cytokines. Aging microglia fail to clear amyloid-beta plaques, accelerates tau-pathology and enhances the chronic neuroinflammation, exacerbating the α-synuclein aggregation. These changes significantly impacted the onset of various neurogenerative disorders such as amyotrophic lateral sclerosis, Parkinson's disease, and Alzheimer's disease etc. However, it is important to note that these microglial aging effects might not be perceived as absolute, due to various limitations such as microglial heterogeneity, intercellular complexity across brain regions and variability in human aging owing to genetic and epigenetic variations. Regardless of this the future perspective of such insights are of immense relevance as novel therapeutic approaches can be formulated if the molecular and cellular mechanisms of aging microglial perturbations are understood. Future research should focus on restoring microglial homeostasis to mitigate the effects of aging on the brain and slowing the progression of neurodegenerative diseases.

Sensory impairment, including hearing and vision impairment, are prevalent in older adults and may significantly contribute to cognitive frailty. However, the longitudinal association between sensory impairment and cognitive frailty has not been fully explored. The aim of our study was to examine such associations in four nationally representative cohorts to inform strategies for preventing cognitive frailty and promoting healthy aging.

Assessing health parameters is essential for understanding the decline of physiological functions during aging. The nematode Caenorhabditis elegans serves as a powerful model in aging research. This paper introduces representative methods for evaluating health parameters using C. elegans and provides quantitative indicators of functional declines during aging. This manuscript will be useful for researchers unfamiliar with healthspan studies using C. elegans and may help perform physiological experiments for healthy aging.

Observational studies examining the relation of body composition with lifespan are inconsistent. We conducted a Mendelian Randomization (MR) study to assess effects of key features of body composition on lifespan.

The global increase in aging populations has heightened the urgency to develop effective interventions for age-related cognitive decline. Skeletal muscle has recently emerged as a potential modulator of brain health, particularly in the context of aging. This study investigates the effects of electrical muscle stimulation (EMS) on cognitive function and neuropathology in Senescence-Accelerated Mouse (SAMP8), a model of aging-associated cognitive decline. SAMP8 mice were divided into 3 groups: healthy controls (SAMR1), untreated SAMP8, and EMS-treated SAMP8. EMS was applied daily for 30 days, and behavioral, histological, and molecular markers were analyzed. Results demonstrated that EMS significantly improved muscle strength and endurance while reducing amyloid-β accumulation and phosphorylated tau (p-Tau) levels in the hippocampus. Furthermore, EMS decreased neuroinflammation and partially restored synaptic plasticity. However, EMS had limited effects on cortical pathology and cognitive function, suggesting that localized brain changes may not fully translate to behavioral improvements. These findings indicate that EMS exerts neuroprotective effects through skeletal muscle activation, providing a potential non-pharmacological intervention for age-related neurodegeneration. Future studies should explore the underlying mechanisms and translational applicability to human dementia treatment.

Aging is a multifactorial biological process characterized by the irreversible accumulation of molecular damage, leading to an increased risk of age-related diseases. With the global prominent rise in aging populations, elucidating the mechanisms underlying the aging process and developing strategies to combat age-related diseases have become a pressing priority. Extracellular vesicles (EVs) have gained significant attention due to their role in intercellular communication. EVs are known for their ability to deliver biocargoes, such as miRNA, proteins, and lipids, implicating their involvement in disease pathogenesis and intervention. In this review article, we explore the dual role of EVs in age-related diseases: contributing to the pathogenesis of diseases by transferring deleterious molecules, while also offering therapeutic ability by transferring beneficial molecules. We also highlight the application of EVs as biomarkers for early diagnosis of age-related diseases, paving the way for early intervention and precision medicine. Additionally, we discuss how analysing the composition of EVs cargo can provide insights into disease progression. Finally, we address the challenges and future perspectives of EV-based-therapy in clinical translation, including standardization of EVs isolation methods and improving cargo specificity.

Peripheral arterial disease (PAD) is prevalent among the elderly, and therapeutic neovascularization is a research hotspot in PAD treatment. Supplementing nicotinamide adenine dinucleotide (NAD

As populations age, understanding cognitive decline and age-related diseases like dementia has become increasingly important. "SuperAgers," individuals over 65 with cognitive abilities similar to those in their 40s, provide a unique perspective on cognitive reserve. This study analyzed 55 blood biomarkers, including cellular components and metabolism/inflammation-related factors, in 39 SuperAgers and 42 typical agers. While conventional statistical analyses identified significant differences in only four biomarkers, advanced feature selection and machine learning techniques revealed a broader set of 15 key biomarkers associated with SuperAger status. A predictive model built using these biomarkers achieved an accuracy of 76% in cognitive domain prediction. To address the limitation of small sample sizes, data augmentation leveraging large language models improved the model's robustness. Shapley Additive exPlanations (SHAP) provided interpretability, revealing the impact of specific blood factors on cognitive function. These findings suggest that certain blood biomarkers are not only associated with cognitive performance but may also serve as indicators of cognitive reserve. By utilizing simple blood tests, this research presents a clinically significant method for predicting cognitive function and identifying SuperAger status in healthy elderly individuals, offering a foundation for future studies on the biological mechanisms underpinning cognitive resilience.

Osteoarthritis, a common age-related joint disease, causes cartilage degeneration, leading to pain and disability. While pain management exists, cartilage regeneration options are limited. Exogenous mitochondria transfer is a novel regenerative approach. This study aimed to investigate the effects of exogenous mitochondrial transfer on cellular function, oxidative stress, inflammation, and apoptosis in osteoarthritic chondrocytes.

Models of personality and health emphasize the role of Five-Factor Model personality traits (particularly neuroticism and conscientiousness) in cognitive health outcomes. A complementary literature on lifecourse models of cognitive aging indicate that earlier life factors (socioeconomic status, adverse childhood experiences, childhood health) are critical for later life cognitive function and dementia risk. There is growing evidence that these earlier life factors also contribute to adult personality. The present review integrates models of personality and health into lifecourse models of cognitive aging and posits personality traits as central mechanisms between earlier life factors and cognitive health. Development of a more detailed lifespan model will help identify chains of risk and resilience to promote better late-life cognition, health, and well-being.

Corn silk, the stigmas and styles of maize, is a traditional medicinal herb with promising anti-aging potential. This review systematically examines its bioactive compounds, including flavonoids, polysaccharides, terpenoids, and sterols, which contribute to its antioxidant, anti-inflammatory, anti-tumor, and metabolic regulatory effects. These properties support its potential therapeutic applications in aging-related conditions such as neurodegenerative diseases (e.g., Alzheimer's and Parkinson's), cardiovascular disorders, metabolic diseases (e.g., diabetes and obesity), and kidney dysfunction. Preclinical studies highlight corn silk's role in mitigating oxidative stress and inflammation-key drivers of aging. Additionally, preliminary clinical trials, particularly on its anti-diabetic effects, show promise, but larger randomized controlled trials are needed to establish its safety and efficacy. Future research should focus on standardizing extraction methods, elucidating mechanisms of action, and exploring synergistic interactions among its bioactive constituents. Together, this review emphasizes the need for continued investigation into corn silk as a natural, multi-target therapeutic agent for aging and age-related diseases, and advancing its potential for clinical applications.

One of the strongest signatures of aging is an accumulation of mutant mitochondrial DNA (mtDNA) heteroplasmy. Here we investigate the mechanism underlying this phenomenon by calling mtDNA sequence, abundance, and heteroplasmic variation in human blood using whole genome sequences from ~750,000 individuals. Our analyses reveal a simple, two-step mechanism: first, individual cells randomly accumulate low levels of \"cryptic\" mtDNA mutations; then, when a cell clone proliferates, the cryptic mtDNA variants are carried as passenger mutations and become detectable in whole blood. Four lines of evidence support this model: (1) the mutational spectrum of age-accumulating mtDNA variants is consistent with a well-established model of mtDNA replication errors, (2) these mutations are found primarily at low levels of heteroplasmy and do not show evidence of positive selection, (3) high mtDNA mutation burden tends to co-occur in samples harboring somatic driver mutations for clonal hematopoiesis (CH), and (4) nuclear GWAS reveals that germline variants predisposing to CH (such as those near TERT, TCL1A, and SMC4) also increase mtDNA mutation burden. We propose that the high copy number and high mutation rate of mtDNA make it a particularly sensitive blood-based marker of CH. Importantly, our work helps to mechanistically unify three prominent signatures of aging: common germline variants in TERT, clonal hematopoiesis, and observed mtDNA mutation accrual.

While memory regulation is predominantly understood as autonomous to neurons, factors outside the brain can also affect neuronal function. In Caenorhabditis elegans, the insulin/IGF-1-like signaling (IIS) pathway regulates longevity, metabolism and memory: long-lived daf-2 insulin/IGF-1 receptor mutants more than double memory duration after a single training session, and it was assumed that memory regulation was strictly neuronal. However, here we show that degradation of DAF-2 in the hypodermis also greatly extends memory, via expression of the diffusible Notch ligand, OSM-11, which in turn activates Notch signaling in neurons. Single-nucleus RNA sequencing of neurons revealed increased expression of CREB and other memory genes. Furthermore, in aged animals, activation of the hypodermal IIS-Notch pathway as well as OSM-11 overexpression rescue both memory and learning via CREB activity. Thus, insulin signaling in the liver-like hypodermis non-autonomously regulates neuronal function, providing a systemic connection between metabolism and memory through IIS-Notch-CREB signaling from the body to the brain.

Geroprotectors are compounds that target the underlying mechanisms of ageing to delay the onset of age-related diseases and extend both lifespan and health span. As ageing is driven by the accumulation of cellular damage, DNA instability, epigenetic changes, mitochondrial dysfunction, and chronic inflammation, the concept of geroprotection focuses on compounds that can mitigate these processes. Oleuropein (OLE) and its derivative hydroxytyrosol (HT), both phenolic molecules derived from Olea europaea (olive tree), have gained significant attention as potential geroprotectors due to their potent antioxidant and anti-inflammatory properties. These phytochemicals, central to the Mediterranean diet, activate key molecular pathways such as nuclear factor erythroid 2-related factor 2, reducing oxidative stress and modulating inflammatory responses. Through these mechanisms, OLE and HT help counteract inflammageing, a critical factor in age-related dysfunction. This review highlights the role of OLE and HT as geroprotective agents, emphasising their ability to target the hallmarks of ageing and their potential to improve health span by slowing the progression of age-related conditions. With proven efficacy in various biological models, these compounds represent promising tools in the ongoing search for strategies to enhance the quality of life in ageing populations.

During tissue repair, stress-induced cellular senescence represents a critical factor that impedes the regenerative potential of tissues. While the regulatory effects of matrix viscoelasticity on cellular behavior have been documented, their role and correlated mechanisms underlying cellular senescence remain unclear. In this study, we engineered a viscoelastic gel matrix exhibiting a storage modulus of approximately 3 kPa, with a tunable loss modulus ranging from 0 to 300 Pa by incorporating linear alginate and modulating the compactness of a polyacrylamide-based covalent network. Utilizing a UV-induced senescence model, we observed that increasing the matrix's viscoelasticity from 0 Pa to 300 Pa led to a significant reduction in the proportion of senescent cells, from 90.5% to 22.7%. Furthermore, cells cultured in these matrices exhibited a tendency to form cell aggregation, with the cell populations demonstrating a collective resistance to stresses. This indicated that viscoelastic materials would promote enhanced cellular interactions, thereby strengthening cellular resilience against UV-induced stresses. Furthermore, combined with microarray analysis, it was concluded that the presence of viscoelastic components activated the connexin 43 (Cx43)-modulated gap junction for cluster formation, thereby suppressing the senescence-associated signaling pathways, including Wnt/β-catenin, MAPK, NF-κB, and TGF-β. Additionally, the integrin-cytoskeleton-Yes-associated protein (YAP) signaling axis played an active role in delaying cell aging. These results provide novel insights into the regulatory role of viscoelastic materials in cellular senescence and offer a compelling foundation for the development of advanced biomaterials for tissue repair.

Early life stresses impact reproductive outcomes in many organisms. In response to crowding and starvation, C. elegans nematodes form dauer larvae, in which development arrests until conditions improve. We discovered dramatic differences in gonad size and germ cell number among dauers that form under different conditions. We used live cell imaging of fluorescent markers in otherwise wild-type and mutant animals combined with food-removal, recovery, and brood-size assays to investigate the causes and consequences of this germline difference. Pre-dauer feeding, but not nutrient sensing via the DAF-2/insulin-like signaling receptor or DAF-7/TGF-{beta}, is required for plasticity in gonad size. Gonad differences in dauer have lifelong reproductive consequences; worms with small dauer gonads recover to have smaller broods. Somatic and germline development are decoupled as pre-dauer starvation induces germline quiescence while the soma continues its development until dauer is formed. A rapid return to germline Notch-dependence and an increase in presentation by the germline stem cell niche of the Notch ligand LAG-2--regulated at the protein and not transcript level--are among the earliest events of dauer recovery.

The transcription factors FOXO4 and p53 regulate aging, and their deregulation has been linked to several diseases, including cancer. Under stress conditions, cellular senescence is promoted by p53 sequestration and senescence-associated protein p21 transcriptional upregulation induced by interactions between the FOXO4 Forkhead DNA-binding domain and the p53 transactivation domain. However, the molecular details of these interactions remain unclear. Here, we report that these interactions between p53 and FOXO4 domains are highly heterogeneous. The p53 transactivation domain primarily interacts with the region formed by the N-terminal helical bundle of the FOXO4 Forkhead domain but retains a substantial degree of flexibility in the complex. In addition, NMR data-driven molecular simulations suggest that p53 interacts with FOXO4 through multiple binding modes. Overall, our findings not only provide the structural insights into interactions between FOXO4 and p53 but also highlight their potential as targets for developing senolytic compounds.

Seed longevity is crucial for both ecological and agronomical value. Previously, we demonstrated that the E3 ligase Arabidopsis tóxicos en levadura 5 (ATL5) positively regulates seed longevity by mediating the degradation of the activator of basal transcription 1 in Arabidopsis. In the present study, we demonstrated that clathrin light chain 2 (CLC2), another ubiquitinated substrate of ATL5, affects seed longevity in Arabidopsis. The interaction between CLC2 and ATL5 was first identified in yeast cells and then in planta. Seeds of clc2 mutants displayed slower accelerated aging, whereas CLC2-OE seeds showed faster accelerated aging than wild-type seeds. In vitro assay showed that ATL5 promotes CLC2 degradation through the 26S proteasome pathway. Degradation of endogenous CLC2 was diminished in atl5 seeds, which could be induced by aging and occurs in a proteasome-dependent manner. Moreover, the role of CLC2 in seed longevity was independent of endocytosis; however, CLC2 exhibited transcriptional activation activity. Transcriptomic analysis revealed that the expression of numerous ribosomal protein genes was significantly upregulated in clc2 seeds after aging. Collectively, our study demonstrated that CLC2 is another ubiquitinated substrate of ATL5 that negatively regulates seed longevity by influencing the expression of ribosomal protein genes in Arabidopsis.

Vascular calcification, an age-associated disorder, is a highly regulated biological process similar to bone formation. Growth differentiation factor 11 (GDF11), a secreted member of the transforming growth factor beta (TGF-β) superfamily, has been shown to act as an anti-aging factor in the brain, heart, skin, and skeletal muscle. Nevertheless, whether GDF11 affects vascular calcification and the underlying mechanisms remain unclear. In the present study, beta-glycerophosphate and calcium chloride-induced calcification of vascular smooth muscle cells (VSMCs) and a VitD

Physical inactivity, which increases the risk of chronic diseases in older adults, is prevalent among older adults in Hong Kong. To address this problem, the Hong Kong government has been proactively promoting active aging.

Social interaction has been shown to prolong lifespan and healthspan. For older adults living alone, social interaction largely comes from formal social participation, and thus depends on the sociability of the individual. This study aims to understand the effect of age on sociability, and the possible mechanisms behind the change. 196 German participants aged 20-77 (Mage = 37.9) completed a series of questionnaires as part of the Leipzig Study for Mind-Body-Emotion Interactions. Sociability was measured by a subscale of the Trait Emotional Intelligence Questionnaire (Short Form), and network-based statistics were performed on resting-state functional connectivity data to identify networks positively and negatively correlated with age. Mediation analysis was carried out between age and sociability, with both sets of edges as mediators. Overall, age correlated negatively with sociability. The brain network correlating positively with age correlated negatively with sociability, and vice versa for the network correlating negatively with. Both networks independently and completely mediated the age-sociability relationship. The limbic-insular and ventral attention-somatomotor connectivity featured prominently in the age-positive network, while the age-negative network is largely represented by subcortical-parietal and frontoparietal-default mode connections. Networks associated with brain aging can explain the negative relationship between age and sociability. Per Dunbar's social brain hypothesis, age-related disruption in intrinsic functional connectivity may impair socio-cognitive functions necessary for forming and maintaining relationships, thus causing a decrease in sociability. Psychoeducation of these changes that occur with natural aging could prove useful in the promotion of successful aging.

The global population of individuals aged 60 years and older now exceeds one billion and continues to grow, underscoring the necessity of addressing cognitive challenges associated with aging. The present study aimed to evaluate the enhancement and suppression of attentional mechanisms, as well as working memory efficiency (WME), across the adult lifespan. Additionally, it examined factors that may moderate these relationships. A total of 194 participants, aged between 20 and 80 years, completed questionnaires and participated in an experimental task designed to assess attentional mechanisms and WME. The results indicated that both enhancement and suppression mechanisms remained stable throughout adulthood; however, WME showed a decline with increasing age. Notwithstanding, several moderating variables influenced these outcomes. Cognitive reserve and current cognitive functioning were found to be positive predictors of WME, whereas depression and anxiety exerted negative effects. Furthermore, a low level of cognitive reserve uniquely predicted diminished suppression of irrelevant information with advancing age. WME was consistently lower across all ages among participants with high levels of depression, in contrast to those with low or moderate levels, in whom WME followed the expected age-related trajectory. Additionally, elevated anxiety levels and an average sleep duration of five hours were associated with reduced WME, particularly in relation to the enhancement mechanism. Conversely, low levels of depression were linked to improved WME via the enhancement mechanism. These findings underscore the importance of moderating factors that may mitigate cognitive decline or support cognitive functioning throughout the aging process.

We conducted a randomized, placebo-controlled trial to assess the safety and biological age (BA) effects of various therapeutic plasma exchange (TPE) regimens in healthy adults over 50. Participants received bi-weekly TPE with or without intravenous immunoglobulin (IVIG), monthly TPE, or placebo. Randomization was based on entry date, and treatments were blinded to maintain objectivity. Primary objectives were to assess long-term TPE safety and changes in biological clocks. Secondary goals included identifying optimal regimens. Exploratory analyses profiled baseline clinical features and longitudinal changes across the epigenome, proteome, metabolome, glycome, immune cytokines, iAge, and immune cell composition. We demonstrate in 42 individuals randomized to various treatment arms or placebo that long-term TPE was found to be safe, with only two adverse events requiring discontinuation and one related to IVIG. TPE significantly improved biological age markers, with 15 epigenetic clocks showing rejuvenation compared to placebo (FDR < 0.05). Biweekly TPE combined with intravenous immunoglobulin (TPE-IVIG) proved most effective, inducing coordinated cellular and molecular responses, reversing age-related immune decline, and modulating proteins linked to chronic inflammation. Integrative analysis identified baseline biomarkers predictive of positive outcomes, suggesting TPE-IVIG is particularly beneficial for individuals with poorer initial health status. This is the first multi-omics study to examine various TPE modalities to slow epigenetic biologic clocks, which demonstrate biological age rejuvenation and the molecular features associated with this rejuvenation. Trial Registration: Registered trial NCT06534450 on clinicaltrials.gov under the purview of the Diagnostic Investigational Review Board.

Aerobic capacity is conventionally equated with cardiorespiratory fitness (CRF), but its physiological essence extends far beyond cardiopulmonary performance. Aerobic capacity is an integrative physiological indicator reflecting the entire process from oxygen uptake and transport to mitochondrial energy conversion, with mitochondrial function constituting its molecular core. Emerging evidence reveals robust associations between diminished aerobic capacity and increased risks of non-communicable chronic diseases and age-related functional decline. However, its potential as a valuable tool for early disease detection and intervention remains undervalued in clinical practice. By synthesizing recent clinical and experimental studies, we highlight the crucial role of aerobic capacity, particularly its mechanistic links to impaired mitochondrial function, which drives disease progression through impaired energy metabolism and chronic inflammation. Furthermore, exercise interventions designed to enhance aerobic capacity have shown promise in improving mitochondrial efficiency, promoting cardiometabolic adaptation, and boosting overall health, thus offering an effective strategy for chronic disease prevention. We advocate for inclusion of aerobic capacity assessments in routine health evaluations and emphasize the need to integrate aerobic capacity optimization into public health frameworks to advance preventive strategies against chronic diseases and promote healthy aging.

Human aging is intrinsically associated with the onset and the progression of several disease states causing significant disability and poor quality of life. Although such association was traditionally considered immutable, recent advances have led to a better understanding of several critical biochemical pathways involved in the aging process. This, in turn, has stimulated a significant body of research to investigate whether reprogramming these pathways could delay the progression of human ageing and/or prevent relevant disease states, ultimately favoring healthier aging process. Cellular senescence is regarded as the principal causative factor implicated in biological and pathophysiological processes involved in aging. Asymmetric dimethylarginine (ADMA) is an endogenous inhibitor of nitric oxide synthase and an independent risk factor for several age-associated diseases. The selective extracorporeal removal of ADMA is emerging as a promising strategy to reduce the burden of age-associated disease states. This article discusses the current knowledge regarding the critical pathways involved in human aging and associated diseases and the possible role of ADMA as a target for therapies leading to healthier aging processes.

Skin ageing is a complex and multifaceted biological process that involves the accumulation of senescent dermal fibroblasts. While fractional microneedle radiofrequency (MNRF) is widely used for skin rejuvenation, the underlying molecular mechanisms are unknown. This study aimed to investigate the efficacy of fractional MNRF in altering the cellular milieu of aged skin and to evaluate clinical skin improvements. Thirty female volunteers aged ≥ 60 years with visible periorbital wrinkles received four consecutive treatments of either microneedling or MNRF on randomly assigned facial sides. Based on biophysical measurements, MNRF treatment improved wrinkles, elasticity, hydration, and transepidermal water loss compared to baseline. Histological analysis revealed that the MNRF-treated sides exhibited increased proliferation of non-senescent fibroblasts, a reduced number of senescent fibroblasts, and elevated collagen and elastin levels, compared to the MN-treated sides. In additional analyses, differences in collagen density and hydration between the two sides of the face were statistically significant only in subjects with a marked reduction in senescent fibroblasts in MNRF-treated sides. Our data suggest that, compared to MN, MNRF induces greater clinical and histological improvements in aged skin, likely by altering the dermal fibroblast milieu through the dual effect of eliminating senescent fibroblasts and increasing the number of non-senescent fibroblasts.

Despite the growing interest in developing anti-aging drugs, high costs and low success rates of traditional drug discovery methods pose significant challenges. Aging is a complex biological process associated with numerous diseases, making the identification of compounds that can modulate aging mechanisms critically important. Accelerating the discovery of potential anti-aging compounds is essential to overcome these barriers and enhance lifespan and healthspan. Here, we present ElixirSeeker, a machine learning framework designed to maximize feature capture of lifespan-extending compounds through multi-fingerprint fusion mechanisms. Utilizing this approach, we identified several promising candidate drugs from external compound databases. We tested the top six hits in Caenorhabditis elegans and found that four of these compounds-including Praeruptorin C, Polyphyllin VI, Thymoquinone, and Medrysone-extended the organism's lifespan. This study demonstrates that ElixirSeeker effectively accelerates the identification of viable anti-aging compounds, potentially reducing costs and increasing the success rate of drug development in this field.

Klotho, named after the youngest of the three Fates in Greek mythology daughters of Zeus and Nyx, who together spin the thread of life, allot destiny, and determine the time of passing for both mortals and immortals, is an important regulatory factor in aging and cancer dynamics. Initially described as an aging-suppressing protein, Klotho is now recognized for its more diverse role in modulating key signaling pathways like Wnt/β-catenin, IGF-1, PI3K/AKT, and TGF-β. Essentially, its various pro-cellular health functions, such as antioxidant, anti-inflammatory, and tumor-suppressive activities, are, in fact, considered that ensures the maintenance of cellular health and reduce complications related to aging. Klotho deficiency is associated with accelerated aging, chronic kidney disease, cardiovascular disorders, neurodegeneration, and various cancers. This review thus covers the twin roles of Klotho as an antiaging and tumor-suppressor protein, on their therapeutic potential, as well as advances in delivery systems and development of biomarkers and challenges for clinical translation.. Moreover, natural strategies like exercise and dietary interventions are explored that could help overcome Klotho deficiency. Further research with Klotho may offer a paradigm shift in the treatment of aging and cancer and add yet another avenue to increase survival of the patients.

Metformin, a medication primarily used to treat diabetes, has gained attentions for its potential antiaging properties. Although the metabolic and cellular pathways behind its longevity effects have been widely studied, few studies have explored the epigenetic regulatory effects of metformin, which are a crucial factor in aging processes. In this study, we examined the antiaging effects of metformin using the Brachionus rotifer as a model, focusing on the regulation of mRNA N6-methyladenosine (m6A), a key RNA modification involved in mRNA stability, translation, and splicing. We found metformin significantly extended the rotifers' lifespan, mimicking the effects of dietary restriction (DR), a well-established antiaging intervention. Both metformin and DR modulate m6A dynamics, with a notable reduction in the m6A modification of MTR (5-methyltetrahydrofolate-homocysteine methyltransferase). This reduction led to decreased MTR expression and lowered levels of S-adenosylmethionine (SAM), a critical metabolite in the one-carbon cycle. We propose that the downregulation of MTR through m6A modification limits methionine synthesis and imposes methionine restriction, a key factor in promoting longevity. Our findings reveal a novel epitranscriptional regulatory model by which metformin and DR modulate m6A to extend lifespan, highlighting MTR as a central regulator of aging and suggesting potential therapeutic strategies for healthy aging through m6A and methionine metabolism.

As societies age, policy makers need tools to understand how demographic aging will affect population health and to develop programs to increase healthspan. The current metrics used for policy do not distinguish differences caused by early-life factors, like prenatal care and nutrition, from those caused by ongoing changes in people's bodies that are due to aging and that may be modifiable. Here we introduce an adapted Pace of Aging method designed to quantify differences between individuals and populations in the speed of aging-related health declines. The adapted Pace of Aging method, implemented in parallel in data from the US Health and Retirement Study and in the English Longitudinal Study of Aging (combined n = 19,045), integrates longitudinal data on blood biomarkers, physical measurements and functional tests. It reveals stark differences in rates of aging between population subgroups and demonstrates strong and consistent prospective associations with incident morbidity, disability and mortality. This adapted and generalizable method to measure Pace of Aging can advance the population science of healthy longevity.

Aerobic exercise training is a potent intervention for the treatment and prevention of age-related metabolic disease, which is characterized by both insulin and glucagon resistance. While insulin resistance is a key driver of metabolic disease in aging, glucagon signaling is equally critical in maintaining both glucose and lipid homeostasis, particularly during exercise. Previous studies have established the glucagon sensitizing effects of exercise training in younger animals. Most studies in rodents employ rigorous and carefully dosed forced exercise protocols. This forced exercise is a stressful paradigm. We implemented a voluntary wheel running (VWR) intervention to assess the effects of aging and exercise training on glucagon sensitivity. We initiated 12-weeks of VWR in young adult (6-month-old) and middle-aged (12-month-old) C57BL/6NCrl male mice. Glucagon sensitivity, as assessed by glucagon stimulated hyperglycemia, was decreased in middle-aged compared to young adult sedentary mice (P=0.046). While VWR did not affect glucose clearance, circulating insulin, glucagon, or insulin sensitivity, regardless of age, VWR improved glucagon responsivity only in middle-aged mice (P=0.031). VWR increased hepatic glycogen content and increased glucagon-stimulated glycogen depletion, regardless of age (P<0.01). Results from these studies suggest that exercise training can enhance liver glucagon action in aging mice without otherwise altering glucose homeostasis.

The proportion of older people has been enhanced in the past half-century around the world, consequently leading to the increasing overall prevalence of age-related health conditions. The present study aims to summarize the current evidence regarding the association between dietary diversity (DD) and the components of healthy aging.

The gut microbiome early in life plays a crucial role in development of the host and affects health throughout life. The definition of a healthy microbiome early in life has not been established, and the underlying mechanism of how a young host selects appropriate microbes for colonization remains unclear. Understanding the mechanism may provide insights into novel preventive and therapeutic strategies by correcting dysbiosis early in life. We employed germ-free mice early in life (4 weeks of age) and later in life (10 weeks of age) for fecal microbiota transfer (FMT) from specific pathogen-free mice. We performed age-unmatched FMT between recipients early in life and donors early or later in life, in addition to common age-matched FMT. Age-matched FMT resulted in significantly different bacterial compositions between recipients early vs. later in life. When the gut microbiome from donors early or later in life was transferred to recipients early in life, bacterial compositions of recipients from donors later in life were similar to those of recipients from donors early in life. This finding suggests that the host early in life has mechanisms to select microbes appropriate for age from the exposed microbiome. We hypothesized that the age-specific intestinal environment promotes age-appropriate intestinal microbiome colonization and examined gene expression in the intestinal mucosa of germ-free mice. We observed that gene expression profiles were different between early vs. later in life. Correlation analysis demonstrated that genera Lachnospiraceae NK4A136 group and Roseburia were positively correlated to genes expressed predominantly early in life, but negatively with genes expressed predominantly later in life. We confirmed that the relative abundance of these genera was significantly higher in specific pathogen-free mice early in life compared with mice later in life. The characteristic gene expression of the intestinal mucosa early in life might play roles in selecting specific bacteria in the intestinal microbiome early in life.

Aging is a pan-organ process with an intricate and multimodal nature. Deciphering the aging phenomenon is complex, yet recent attention is analyzing the potential benefits of non-invasive life adjustments to achieve healthy aging. Omega-3 polyunsaturated fatty acids (FA) have emerged as promising nutraceuticals for a plethora of different medical conditions. In the current study we conducted an in-depth, bottom-up, global, shotgun proteomic study (LC-MS/MS) investigating both the effects of aging on skeletal muscle and the potential alterations due to ω-3 FA. Sprague Dawley rats were fed different diets and divided into four groups (n = 5 per group): adult controls (7-8 months, ADCTL); aged controls (22 months, AGCTL); and adult (ADω3) and aged (AGω3) rats fed an ω-3 supplemented diet. Among the identified 30,000 soleus proteins, our proteomic analysis identified 149 proteins differentially expressed in aging; 207 proteins with aging, but fed ω-3 FA; and 105 and 26 proteins, respectively, when aged and adult rats were fed ω-3 FA. Aging alone (ADCTL/AGCTL) affects many processes: carbohydrate and lipid metabolisms, proteostasis, mRNA processing and sarcomeric proteins. With FA supplementation and aging (ADω3/AGω3) similar processes were affected, but increased chromatin-related protein abundances (methylation or histone deacetylation) were observed in AGω3; while proteins involved in OXPHOS and mitochondrial homeostasis, including mTOR, were more represented in ADω3 rats. Supplementation with FA had a greater effect in aged rats (AGCTL/AGω3) than in adult ones (ADCTL/ADω3). In the ADCTL/ADω3 comparison, modest changes were seen, whereas in the AGCTL/AGω3 comparisons DNA damage repair increased and protein synthesis and degradation were observed. Further, a potential link to enhancement of myogenesis is also evident. The data presented in this work suggest potential beneficial and protective effects of ω3 FA supplementation in the soleus muscle, as well as some potential molecular mechanisms of action.

Age-related muscle loss can be decreased with increased protein intake. Recent evidence suggests that increasing animal-based protein such as lean beef can be the most effective for age-related muscle repair and growth. Culinary medicine (CM) is a science-based field to teach people the art of food and cooking with the science of medicine to improve health.

The prevalence of metabolic dysfunction-associated steatotic liver disease (MASLD) is rapidly increasing and is caused by excessive fat deposition in the liver due to non-alcoholic factors. Aging is a major risk factor for the development and progression of MASLD. In this study, we investigated the metabolic effects of tilorone, a synthetic small molecule, in a high-fat diet (HFD) mouse model, with a focus on the liver function and signaling. We demonstrate that tilorone attenuated HFD-induced steatosis by restoring bone morphogenetic protein 9 (BMP9)-Smad1/5/8 signaling and upregulating peroxisome proliferator-activated receptor gamma (PPARγ) expression. Tilorone reduced HFD-induced increases in body weight, adipose tissue and liver weight, and blood glucose levels, and improved glucose tolerance in HFD mice. PET/MRI imaging demonstrated enhanced

Age-related degenerative changes in intervertebral discs (IVDs) can lead to lower back pain, and even paralysis. This topic is therefore garnering growing attention in an increasingly ageing society. The oxidative stress-induced degenerative process is a major contributor to apoptosis in nucleus pulposus cells. However, the regulatory mechanism of NAD-dependent protein deacetylase Sirtuin-1 (SIRT1) on apoptosis in oxidative stress-induced rat nucleus pulposus cells remains unclear.

Aging is a significant risk factor for cardiovascular diseases, with ischemic heart disease (IHD) being the leading cause of cardiovascular-related mortality. Inhibition of FOXO4, which selectively eliminates senescent cells, offers protective effects on the aging myocardium. However, the removal of senescent cells may lead to a reduction in tissue cell density, thereby exacerbating tissue space formation and perivascular fibrosis. Therefore, selectively eliminating senescent cells in the aging heart, while simultaneously replenishing therapeutic bone marrow-derived mesenchymal stem cells (BMSCs), holds substantial therapeutic potential for synergistically combating cardiac aging. This study proposes a promising cardiac rejuvenation strategy using ultrasound-targeted microbubble destruction (UTMD)-mediated delivery of shFOXO4/SDF1 to eliminate cellular senescence and enhance BMSC homing. Transcriptomic analysis identified FOXO4 as a pivotal transcription factor in cardiac aging, with FOXO4 protein predominantly expressed in cardiac fibroblasts (CFs) and vascular endothelial cells in the myocardium of aged rats. Knockdown of FOXO4 in aging CFs reversed cellular senescence, and co-culturing these rejuvenated CFs with BMSCs further enhanced the reversal of senescence and bolstered resistance to oxidative stress. The use of UTMD for delivering shFOXO4/SDF1 in dual-gene therapy significantly enhanced BMSC homing, ameliorating cardiac aging, oxidative stress, and inflammation. In an ischemia-reperfusion injury (MIRI) model, pretreatment with shFOXO4/SDF1 effectively reduced cardiomyocyte apoptosis, promoted neovascularization, reduced infarct size, and improved cardiac function. The combined removal of senescent cells and enhanced BMSC homing synergistically ameliorated cardiac aging and improved post-MIRI prognosis in aging hearts. These findings provide novel insights and potential therapeutic strategies for addressing cardiac aging and age-related heart diseases.

Mass spectrometry imaging (MSI) is a rapidly advancing technology that provides mapping of the spatial molecular landscape of tissues for a variety of analytes. Matrix-assisted laser desorption/ionization (MALDI)-MSI is commonly employed, however, confident in situ identification and accurate quantification of analytes remain challenging. We present a novel imaging methodology combining trapped ion mobility spectrometry (TIMS)-based parallel accumulation-serial fragmentation (PASEF) with MALDI ionization for targeted imaging parallel reaction monitoring (iprm-PASEF). We investigated the spatial distribution of lipids and metabolites in liver tissues from wild-type and CD38 knockout mice (CD38-/-). CD38, an enzyme involved in nicotinamide adenine dinucleotide (NAD) metabolism, significantly influences liver metabolic function and contributes to age-related NAD decline. Although CD38 deletion previously was linked to improved metabolic phenotypes, the underlying spatial metabolic mechanisms are poorly understood. The spatial iprm-PASEF workflow enabled confident identification and differentiation of lipid isomers at the MS2 fragment ion level and revealed increased NAD+ and decreased adenosine diphosphate ribose (ADPR), a by-product of NAD+ hydrolysis, in CD38-/- livers. This approach provided confident, specific, and robust MS2-based identification and quantification of fragment ions in spatial MSI experiments. Additionally, the innovative iprm-PASEF opens unprecedented opportunities for spatial metabolomics and lipidomics, offering spatially resolved insights into molecular mechanisms.

Aging is a complex biological process associated with nearly all diseases. Alternative splicing is increasingly recognized as an important contributor to aging and a key research pathway for extending human lifespan. In this review, we highlight the findings of alternative splicing in the hallmarks of aging including key processes such as genomic instability, telomere length, protein stability, autophagy processes, etc., as well as antagonistic hallmarks of aging such as various metabolic signals, energy metabolism, clearance of senescent cells, stem cell self-renewal, cell communication and inflammatory process, etc. We also discuss the roles of alternative splicing in age-related diseases, including neurodegenerative diseases, cardiovascular diseases, skeletal muscle-related diseases, metabolic disorders, cancer, sensory degeneration, and chronic inflammation, etc. These studies suggest that new anti-aging therapies could be developed by regulating key splicing proteins or specific splicing events.

A major challenge in aging research is identifying interventions that can improve lifespan and health and minimize toxicity. Clinical studies cannot usually consider decades-long follow-up periods, and therefore, in-silico evaluations using omics-based surrogate biomarkers are emerging as key tools. However, many current approaches train predictive models on observational data, rather than on intervention data, which can lead to biased conclusions. Yet, the first classifiers for lifespan extension by compounds are now available, learned on intervention data. Here, we review evaluation methodologies and we prioritize training on intervention data whenever available, highlight the importance of safety and toxicity assessments, discuss the role of standardized benchmarks, and present a range of feature processing and predictive modeling approaches. We consider linear and non-linear methods, automated machine learning workflows, and use of AI. We conclude by emphasizing the need for explainable and reproducible strategies, the integration of safety metrics, and the careful validation of predictors based on interventional benchmarks.

Alzheimer's disease (AD) is the primary cause of dementia in older individuals, exhibiting an increasing incidence worldwide. Endoplasmic reticulum (ER) quality control has been receiving attention in the pathology of AD. The accumulation of misfolded proteins in the ER will activates the unfolded protein response (UPR), leading to cellular apoptosis. Using a mouse model of 5xFAD, we found that physical exercise (PE) promoted the clearance of ER fragments and inhibited the ER stress via ER autophagy (ER - Phagy). As a result, physical exercise reduced Aβ deposition, inhibited the neuronal apoptosis, ameliorated the emotional and cognitive impairments. Mechanistically, these PE related effects may be linked to the increased FAM134B, an ER-Phagy receptor, by up-regulating neuronal progranulin (Pgrn). Recombinant Pgrn injection could mimic the protective effects of PE, whereas down-regulation of Pgrn could abolish the PE-associated protection. Our findings indicate that physical exercise, as a readily accessible lifestyle intervention, holds significant potential for preventing diseases related to the global aging population.

Although changes in mitochondrial morphology consistently associated with the aging of hematopoietic stem cells (HSCs), the specific molecular and cellular mechanisms involved are partially unclear. Live-cell super-resolution (SR) microscopy has been used to identify distinct HSC subsets that characterized by mitochondria unique morphologies and spatial distributions. The integration of SR microscopy with single-cell RNA sequencing enabled the classification of approximately 200 HSCs from young and aged mice into 5 discrete clusters. These clusters displayed molecular profiles that corresponded to the observed mitochondria states. An integrated approach combining RNA biomarker analysis and potential regulon assessment revealed previously unrecognized roles of GDF15 in mediating mitochondrial signals and morphologies that influence HSC fate. Thus, combining SR imaging with a bioinformatics pipeline provides an effective method for identifying key molecular players in specific phases of cellular transition, even with a relatively small dataset.

The global increase in life expectancy has sparked growing interest in the factors that contribute to exceptional longevity. Between 1990 and 2015, the number of centenarians worldwide more than quadrupled. This study aimed to analyse the relationship between blood-based biomarkers and the likelihood of reaching 100 years of age in Catalonia (2015-2022), and to examine how biomarker variations during COVID-19 affected longevity. Using a retrospective cohort study based on primary care electronic health records from Catalonia, we compared centenarians with individuals aged 92 or older who died before reaching 100 years of age. We analysed anaemia, cholesterol, glycemia, kidney function, and liver function biomarkers. We employed multiple strategies to control for confounding including matching without replacement, adjusting for both observed confounders at both the individual and contextual level, and unobserved confounders, in particular spatial dependence. Our findings reveal that centenarians exhibit higher rates of chronic conditions, greater socioeconomic disadvantage, and increased neighbourhood inequality in urban areas. Biologically, longevity was linked to intermediate levels of ferritin and cholesterol, alongside lower glucose, creatinine, and uric acid levels. Glycaemic balance, indicated by HbA1c and fasting glucose, emerged as a key factor in survival to extreme old age. Additionally, biomarker improvements during the pandemic correlated with an increased likelihood of reaching centenarian age. These results emphasize the complex interplay between biological, behavioural, and contextual factors in determining longevity. While biomarkers provide valuable insights, they are insufficient indicators of healthy ageing. Future research should integrate multiple dimensions, among them, environmental, and social determinants for uncovering the mechanisms of longevity.

The phenomenon of inflammaging, characterized by an increase in low-grade chronic inflammation, is closely associated with diseases related to liver dysfunction. This study investigated daily plasma exchange between 5-week-old and 24-month-old Sprague Dawley rats for 30 days, focusing on protein secondary structures, NLRP3 inflammasome, and necroptosis. Conformation changes in protein secondary structures were identified by infrared spectroscopy-based pattern recognition analysis. Liver biopsies with histochemical and immunohistochemical staining were used to assess molecules associated with inflammation, necroptosis and NLRP3 inflammasome complex. Expression levels of NLRP3 components were determined by qPCR. Enhanced random coils, 3