Anti-aging Research Articles

The Effect of Axenic Dietary Restriction on the Age-Related Changes in Caenorhabditis elegans.

Axenic dietary restriction (ADR) is highly effective in extending lifespan of Caenorhabditis elegans, but its effects on healthspan improvement are less well characterized. Using transmission electron microscopy, morphometric analyses, and functional assays, we found ADR can preserve tissue ultrastructure, including the cuticle, epidermis, and intestinal lumen, and reduce age-associated pathologies like gonad degeneration, uterine tumor clusters, pharyngeal deterioration, and intestinal atrophy. However, there was no notable improvement in behavioral and functional metrics. Our results underscore that lifespan extension through ADR does not inherently translate to broad healthspan improvements.

A mitochondrial unfolded protein response-independent role of DVE-1 in longevity regulation

The special AT-rich sequence-binding (SATB) protein DVE-1 is widely recognized for its pivotal involvement in orchestrating the retrograde mitochondrial unfolded protein response (mitoUPR) in C. elegans. In our study of downstream factors contributing to lifespan extension in sensory ciliary mutants, we find that DVE-1 is crucial for this longevity effect independent of its canonical mitoUPR function. Additionally, DVE-1 also influences lifespan under conditions of dietary restriction and germline loss, again distinct from its role in mitoUPR. Mechanistically, while mitochondrial stress typically prompts nuclear accumulation of DVE-1 to initiate the transcriptional mitoUPR program, these long-lived mutants reduce DVE-1 nuclear accumulation, likely by enhancing its cytosolic translocation. This observation suggests a cytosolic role for DVE-1 in lifespan extension. Overall, our study implies that, in contrast to the more narrowly defined role of the mitoUPR-related transcription factor ATFS-1, DVE-1 may possess broader functions than previously recognized in modulating longevity and defending against stress.

Senolytics Enhance the Longevity of Caenorhabditis elegans by Altering Betaine Metabolism.

Aging triggers physiological changes in organisms that are tightly linked to metabolic changes. Senolytics targeting many fundamental aging processes are currently being developed. However, the host metabolic response to natural senescence and the molecular mechanism underlying the antiaging benefits of senolytics remain poorly understood. In this study, we investigated metabolic changes during natural senescence based on the Caenorhabditis elegans model and pinpointed potential biomarkers linked to the benefits of senolytics. These results suggest that age-dependent metabolic changes during natural aging occur in C elegans. Betaine was identified as a crucial metabolite in the natural aging process. We explored the metabolic effects of aging interventions by administering 3 antiaging drugs-metformin, quercetin, and minocycline-to nematodes. Notably, betaine expression significantly increased under the 3 antiaging drug treatments. Our findings demonstrated that betaine supplementation extends lifespan, primarily through pathways associated with the forkhead box transcription factor (FoxO) signaling pathway, the p38-mitogen-activated protein kinase (MAPK) signaling pathway, autophagy, the longevity regulating pathway, and the target of rapamycin (mTOR) signaling pathway. In addition, autophagy and free radicals are altered in betaine-treated nematodes. Overall, we found that betaine is a critical metabolite during natural aging and that senolytics extend the lifespan of nematodes by increasing betaine levels and promoting autophagy and antioxidant activity. This finding suggests that betaine could be a novel therapeutic target for promoting longevity.

Characterization and comparation of toxicity between natural realgar and artificially optimized realgar

ObjectiveRealgar possesses important medical properties. This article aims to evaluate realgar and emerging artificially optimized realgar to ensure safe clinical use.MethodsMultiple techniques were employed to test natural realgar and artificially optimized realgar. Soluble arsenic content in representative samples were measured. Natural realgar and artificially optimized realgar were administered to KM mice via gavage for 28 days, and the extent of liver and kidney tissue damage, arsenic accumulation and form of arsenic were measured.ResultsNatural realgar and artificially optimized realgar can be distinguished by their physical properties or spectral signatures. ICP-MS and EPMA identified different contents of elements between two groups. In simulated gastric and intestinal fluids, only As (III) and As (V) were detected. Toxicity experiments in vivo demonstrate that both groups caused minimal liver and kidney damage at a dose of 30 mg·kg−1. At a dose of 180 mg·kg−1, artificially optimized realgar caused significantly greater liver and kidney damage.ConclusionThe differences between natural realgar and artificially optimized realgar were successfully distinguished through several methods. In vitro experiments showed that As is the main component exerting their medicinal effects. In vivo toxicity tests demonstrated that at higher dose, artificially optimized realgar exhibited significantly higher toxicity, suggesting that natural and artificially optimized realgar have different toxic properties.

What matters most to healthy older adults in treatment decision making: a discrete choice experiment

Abstract Introduction Shifting to a patient centered approach is crucial for a sustainable healthcare system. Understanding individuals’ priorities is essential for personalized care, especially in critical illness. Treatment decisions are often aimed at disease-specific outcomes such as life extension or survival, while patient-specific outcomes like maintaining independence may be equally vital. How healthy older adults prioritize treatment outcomes in case of a critical illness is currently unknown. Enhanced insight into their priorities can refine healthcare policy, for more personalized care. This study investigated outcomes (attributes) prioritized in hypothetical decision-making by healthy adults over 50 yr. Methods We conducted a Discrete Choice Experiment with individuals aged ≥50, comparing six pairs of hypothetical treatments with five attributes: life expectancy, independence, pain, memory complaints and societal costs. Attribute utility was analyzed using a conditional logit model, and latent class analyses were employed to explore preferences in groups. Results In 333 participants (mean age 70 +/- 7.7) all attributes emerged as impactful with independence being strongest (p < 0.05). Increased life expectancy had positive utility, but only for a two-year increase. Negative utility was found for all other attributes except mild pain and memory complaints. Latent class analyses identified two groups (22.4% and 77.6%). Group one valued life expectancy more (p .019 vs .940), whereas the second group valued independence (p .058 vs.<.001 and societal costs (p.053 vs. <.001) more. Conclusions In a hypothetical case of life-threatening disease, for people aged ≥ 50 life expectancy was not the sole relevant outcome for treatment decisions; pain, independence and societal costs are important and should be discussed in clinical decision making or healthcare policy. Key messages • Older adults find patient-specific outcomes relevant for decision making. • Societal costs are found relevant for decision making for treatment by older adults.

Effects of Target of Rapamycin and Phosphatidylinositol 3-Kinase Inhibitors and Other Autophagy-Related Supplements on Life Span in y w Male Drosophila melanogaster

Various dietary supplements have been shown to extend the life span of Drosophila melanogaster, including several that promote autophagy, such as rapamycin and spermidine. The goal of the study presented here was to test numerous additional potential anti-aging supplements, primarily inhibitors of the target of rapamycin (TOR) and/or phosphatidylinositol 3-kinase (PI3K). Using a single, comparatively long-lived y w test strain, screening was performed in male flies supplemented either throughout adulthood or, in a few cases, beginning in middle or late adult life, with concentrations spanning 4–6 orders of magnitude in most cases. Supplementation with PP242 and deferiprone, an iron chelator, beginning in late adult life had no positive effect on life span. Lifelong supplementation with Ku-0063794, LY294002, PX-866-17OH, Torin2 and WYE-28 had no effect at any dose. Rapamycin, spermidine and wortmannin all had significant life-shortening effects at the highest doses tested. AZD8055, PI-103 hydrochloride and WYE-132 yielded slight beneficial effects at 1–2 doses, but only 100 nM AZD8055 was confirmed to have a minor (1.3%) effect in a replicate experiment, which was encompassed by other control groups within the same study. These compounds had no effect on fly fecundity (egg laying) or fertility (development of progeny to adulthood), but equivalent high doses of rapamycin abolished fertility. The solvent DMSO had no significant effect on life span at the concentrations used to solubilize most compounds in the fly medium, but it drastically curtailed both survival and fertility at higher concentrations. 2-Hydroxypropyl-β-cyclodextrin also failed to extend the life span when provided throughout adulthood or beginning in mid-adult life. Collectively, the results suggest that inhibition of the TOR/PI3K pathway and autophagy through dietary intervention is not a straightforward anti-aging strategy in Drosophila and that further extension of life is difficult in comparatively long-lived flies.

The NAD+ precursor nicotinamide riboside protects against postovulatory aging in vitro.

Postovulatory aging (POA) of oocytes is clinically significant as it mirrors the degeneration observed in maternally aged oocytes, leading to substantial impairments in oocyte quality and the success rates of artificial reproductive technology (ART). The molecular alterations associated with POA, such as the degeneration of the first polar body, an increase in perivitelline space, reactive oxygen species (ROS) accumulation, energy depletion, and chromosomal and DNA damage, underscore the urgency of finding interventions to mitigate these effects. This study aims to identify whether nicotinamide riboside (NR) can prevent POA during the process of in vitro culture and raise the success rates of ART. Taking advantage of an in vitro postovulatory oocyte aging model, we examined the morphological integrity and NAD+ levels of ovulated mouse MII oocytes after 24h of culturing. Following in vitro fertilization, we assessed the embryonic developmental potential of oocytes affected by POA. Using immunofluorescence and confocal microscopy, we measured the levels of ROS, mitochondrial function, and γH2AX. We also evaluated spindle assembly and chromosome alignment. Additionally, we detected the distribution of cortical granules to assess the metabolic and quality changes in POA oocytes with the supplementation of NR. To further our analysis, quantitative real-time PCR was conducted to measure the mRNA expression levels of antioxidant enzymes Sod1 and Gpx1 in the oocytes. With 200μM NR supplementation during in vitro culture for 24h, the oocytes from POA demonstrated reduced signs of aging-related decline in oocyte quality, including reduced ROS accumulation, improved mitochondrial function, and corrected mis-localization of cortical granules. This improvement in oocyte quality is likely due to the inhibition of oxidative stress via the NAD+/SIRT1 signaling pathway, which also helped to restore normal spindle assembly and chromosome alignment, as well as reduce the elevated levels of γH2AX, thereby potentially enhancing the embryonic development potential. Current research provides evidence that NR is an efficient and safe natural component that prevents the process of POA and is thus a potential ideal antiaging drug for raising the success rates of ART in clinical practice.

Dietary pro-oxidant therapy by a vitamin K precursor targets PI 3-kinase VPS34 function.

Men taking antioxidant vitamin E supplements have increased prostate cancer (PC) risk. However, whether pro-oxidants protect from PC remained unclear. In this work, we show that a pro-oxidant vitamin K precursor [menadione sodium bisulfite (MSB)] suppresses PC progression in mice, killing cells through an oxidative cell death: MSB antagonizes the essential class III phosphatidylinositol (PI) 3-kinase VPS34-the regulator of endosome identity and sorting-through oxidation of key cysteines, pointing to a redox checkpoint in sorting. Testing MSB in a myotubular myopathy model that is driven by loss of MTM1-the phosphatase antagonist of VPS34-we show that dietary MSB improved muscle histology and function and extended life span. These findings enhance our understanding of pro-oxidant selectivity and show how definition of the pathways they impinge on can give rise to unexpected therapeutic opportunities.

Determination of 3-hydroxyanthranilic acid in the sweat of healthy older adults

3-Hydroxyanthranillic acid (3HAA) is one of the key metabolites from the tryptophan (TRP) metabolism pathway and is associated with aging, age-associated diseases, and healthy lifespan extension. This study aims to detect 3HAA in the sweat of healthy older adults using simple, high-performance liquid chromatography (HPLC) method. Chromatographic separation using 20 mmol/L sodium acetate, 3 mmol/L zinc acetate, and 7% (v/v) acetonitrile as mobile phase is possible to simultaneous detect 3HAA, KYN, and TRP with UV and fluorescence detection, respectively, under 6 min. This method demonstrated excellent linearity with coefficient of determination (r2) greater than 0.998 for all analytes. The linear range were 0.05–6 µg/mL for TRP, 0.1–6 µg/mL for KYN and 0.2–6 µg/mL for 3HAA. Percentage recoveries from spiked in human sweat ranged from 90 ± 7–101 ± 3 for TRP, 86 ± 1–92 ± 3 for KYN, and 96 ± 1–103 ± 4 for 3HAA. The precision (%RSD) of repeatability and reproducibility is less than 3% and 6%, respectively. This method was used in a cross-sectional study with 81 participants aged 50–79 years, selected randomly from a local primary healthcare hospital’s sampling frame. A detectable amount of 3HAA was observed in all sweat samples, marking the first report of 3HAA presence in human sweat. Additionally, the results revealed that the 3HAA sweat levels increased with age analyzed in three different age groups ranging from 50–59, 60–69, and 70–79. These findings enhance our understanding of sweat profiles and their correlation with aging, potentially further improving early diagnosis, disease monitoring, and development of customized treatment programs for older adults.

Effect of Different Chemical Pretreatments and Packaging Systems on Quality Characteristics of Fresh-Cut Pomegranate Arils (Punica granatum, L.)

Aims: Pomegranate (Punica granatum) is considered as challenging to consume due to its hard structure, hence used for fresh-cut production. Extending the shelf life of fresh-cut pomegranate arils would improve their marketability, transport efficiency, and reduce post-harvest losses. Study Design: Completely Randomized Design by using GRAPES (General R based Analysis Platform Empowered by Statistics). Place and Duration of Study: College of Agriculture, Kerala Agricultural University, February 2022-July 2024. Methodology: Pomegranate arils were extracted and treated with different pre-treatment solutions viz., 0.5% ascorbic acid, 0.5% citric acid, 1% calcium chloride and 1% calcium ascorbate for 4 minutes. The best pre-treatment solution was selected and arils were packaged in different packaging systems viz., shrink wrapping in 15µ polyolefin film, vacuum packaging in laminated pouches, MAP with N2 flushing in laminated pouches, aluminium tray wrapped with cling film and stored under two different storage conditions, after treating with 1% calcium ascorbate. Results: 1% calcium ascorbate was selected as the best pre-treatment solution and pre-treated arils kept in MAP with N2 flushing in laminated pouches stored under low temperature condition was found to be the best packaging and storage system. 1% calcium ascorbate treated arils in MAP with N2 flushing in laminated pouches stored under low temperature condition (10±5oC) had a shelf life of 6 days with superior physiological (PLW, percent leakage), chemical (acidity, vitamin C, phenol) and sensory characters (color, flavour, texture, appearance, taste and overall acceptability) as compared to control (2.00 days). Maximum vitamin C content of 23.02 mg 100g-1 and least phenol content of 214.16 mg 100g-1 were recorded on 6th day of storage. Conclusion: Pomegranate arils pre-treated with 1% calcium ascorbate for 4 minutes, kept in MAP with N2 flushing in laminated pouches stored under low temperature condition (10±5oC) can be considered as a protocol for shelf life extension of pomegranate arils.

The role of high mobility group proteins in cellular senescence mechanisms

Aging is a universal physiological phenomenon, and chronic age-related diseases have become one of the leading causes of human mortality, accounting for nearly half of all deaths. Studies have shown that reducing the incidence of these diseases can not only extend lifespan but also promote healthy aging. In recent years, the potential role of non-histone high-mobility group proteins (HMGs) in the regulation of aging and lifespan has attracted widespread attention. HMGs play critical roles in cellular senescence and associated diseases through various pathways, encompassing multi-layered mechanisms involving protein interactions, molecular regulation, and chromatin dynamics. This review provides a comprehensive analysis of the interactions between HMG family proteins and senescence-associated secretory phenotype (SASP), chromatin structure, and histone modifications, offering a deeper exploration of the pivotal functions and impacts of HMGs in the aging process. Furthermore, we summarize recent findings on the contributions of HMG proteins to aging and age-related diseases. HMG proteins not only regulate senescence-associated inflammation through modulating the SASP but also influence genomic stability and cell fate decisions via interactions with chromatin and histones. Targeting HMG proteins holds great potential in delaying the progression of aging and its associated diseases. This review aims to provide a systematic overview of HMG proteins’ roles in aging and to lay a solid foundation for future anti-aging drug development and therapeutic strategies. With the advancing understanding of the mechanisms by which HMGs regulate aging, developing therapeutic interventions targeting HMGs may emerge as a promising approach to extending lifespan and enhancing healthspan.

Potential probiotic Lactiplantibacillus plantarum DS1800 extends lifespan and enhances stress resistance in Caenorhabditis elegans model

Probiotics are live microorganisms that provide health benefits when administered in appropriate amounts by improving or restoring the balance of intestinal microbiota. Various functional probiotic products have been developed due to the growing interest in the health-promoting and anti-aging effects of enhancing the gut microbiome. Lactiplantibacillus plantarum species are known for their potential to extend lifespan. However, this activity is strain or isolation source specific, necessitating the identification of individual strain functionalities. This study used the C. elegans model to screen probiotics for life-extension effects and analyze their functions. The 43 lactic-acid bacteria strains isolated from fermented foods, breast milk, and human feces were subjected to longevity assays, and L. plantarum DS1800 was selected to demonstrate the most effective lifespan extension. The average lifespan of Caenorhabditis elegans fed DS1800 increased by 17.36% compared with those fed Escherichia coli OP50. Further analysis of the expression of key genes related to longevity revealed the high expression of the skinhead-1 (skn-1), antibacterial, and heat stress resistance genes via the p38 MAPK pathway. These expression patterns suggest that DS1800 extends the lifespan of C. elegans by enhancing its stress resistance and protecting it against pathogens. Additionally, DS1800 exhibited excellent intestinal adhesion, with 7.56% adhesion to HT-29 cells. Therefore, L. plantarum DS1800 is effective in extending the lifespan of C. elegans and can be used as a functional probiotic.

From Critical Raw Materials to Circular Raw Materials.

This perspective addresses the key challenge of advancing the use of Critical Raw Materials (CRMs) and proposes a transition towards circular raw material management. In the context of our current economy, the unsustainable consumption, environmental degradation, geopolitical risks, and economic vulnerabilities associated with CRMs highlight the limitations in ensuring long-term CRM availability, emphasizing the environmental, social, and economic implications. In response, this perspective underlines a multifaceted technological approach to mitigate CRM criticality, focusing on reducing CRM use, substituting CRMs with less critical materials, and enhancing recovery and recycling processes, with Design for Circularity as the most impactful solution. The latter advocates for a paradigm shift in product design and material utilization, emphasizing principles like modular design, product life extension, and the transition from product ownership to service models. Such a holistic approach is not only crucial for sustainable CRM management, but is also key to fostering a resilient and low-carbon economy.

Enhanced Antimicrobial and Degradable Properties of Silver Nanoparticles‐Reinforced Chitosan‐Indian Gooseberry Films for Sustainable Food Packaging

AbstractThis study focuses on creating a biodegradable film composed of chitosan, Indian Gooseberry (IG) extract, and silver nanoparticles (AgNPs), designed to enhance antimicrobial properties and improve food preservation. The presence of AgNPs and IG extract not only dramatically exhibits 99% efficiency in the antimicrobial activity against Staphylococcus aureus (ATCC 6538P), but also achieves this within a remarkably short time interval of ≈20 min. This potent antimicrobial action is attributed to the synergistic effect between the bioactive compounds in the IG extract and the nanoscale silver particles, which together create a highly hostile environment for microbial growth. Moreover, this synergy contributes to 313.94% water vapor transmittance rate and the complete biodegradability of the composite film (93.42% at the end of the second month), ensuring that it leaves no harmful residues in the environment post‐use. The reinforced film exhibits improved hydrophobicity (55.43%), which is crucial for protecting food products from moisture and extending their shelf life. Specifically, the film has been tested as a packaging material for bread, showing a significant extension in shelf life by effectively preventing mold growth and retaining freshness for an extended period (14 days).

Investigation of surface structuring and oxidation performance of Inconel 718 superalloy by laser remelting with different patterns

The current paper deals with the effect of CO2 laser remelting on the surface structuring and oxidation performance of Inconel 718 superalloy, pointing out different laser-induced surface patterns that make a difference in their oxidation properties vital for high-temperature applications. Surface roughness, microstructure, and oxidation behavior characterization were performed using 3D optical profilometry, SEM, and XRD techniques. The oxidation tests conducted at 1000 °C for 24 h have revealed that remelted surfaces offered better oxidation resistance compared to untreated samples. Notably, the patterned sample showed the lowest weight gain under oxidation, and a parabolic regime in oxidation occurred after 100 min; while in an untreated reference sample, this appears only after 200 min. This improvement in performance results from the formation of a chromium-rich oxide layer on the melt pools, which acts as an effective barrier against further oxidation. The findings show that laser remelting, especially with grid-like surface patterns, results in an improvement in both durability and high-temperature performance of Inconel 718; therefore, it is apparently a very promising technique for lifespan extension of industrially relevant materials.

Probabilistic risk assessment method considering machining-induced random residual stress

The aero-engine disks inevitably have manufacturing-induced anomalies and machining-induced random residual stress (RS) in localized and critical areas, which cause a severe threat to the safety of the aircraft. Traditional structural design of the disks fails to establish a quantitative correlation between the machining process and the failure risk. Therefore, this paper proposes a probabilistic model considering random RS to quantify the influence of machining RS subjected to low-cycle fatigue. The RS dispersion is quantified using a scaling parameter, obtained by X-ray diffraction measurements and orthogonal cutting simulations. The crack life database under varying RSs is established for efficient probability calculations. Results show that the coefficient of variation (COV) of the RS on the same machined surface with the same processing parameters is 7.62 % in the local area and 13.74 % in the whole machined surface. The risk results show that the probability of failure (POF) considering the deterministic RS is 2–4 % lower than the POF without RS, owing to the extension of fatigue life by compressive RS. Furthermore, the POF considering the random RS is almost the same (difference <0.6 %) as the POF considering the deterministic RS because the depth of the machining RS is around 0.2 mm. The proposed method predicts the POF more accurately and is thus valuable for the safety assessment of an aero-engine titanium disk.

SlMYB72 interacts with SlTAGL1 to regulate the cuticle formation in tomato fruit.

The cuticle is the first physical barrier covering the surface of tomatoes and plays an important role in multiple stress responses. But the molecular regulatory networks of cuticle formation are not fully understood. In this study, we found that SlMYB72 can interact with SlTAGL1 to regulate the formation of fruit cuticle in tomato. Downregulating the expression of SlMYB72 inhibits the formation of fruit cuticle, resulting in a reduced fruit cuticle thickness, accelerated postharvest water loss, and increased susceptibility to Botrytis cinerea. RNA sequencing analysis showed that downregulation of the SlMYB72 gene decreased the expression levels of genes related to fatty acid and cuticle metabolism. SlMYB72 regulates the cuticle formation by directly binding to the promoter of long-chain acyl-coA synthetases (SlLACS1) and medium-chain alkane hydroxylase (SlMAH1). Moreover, SlMYB72 interacts with SlTAGL1, which can enhance the transcriptional activation of SlMYB72 on the SlMAH1 promoter. Overall, our study expands our understanding of the regulation of cuticle formation by SlMYB72 and provides new insights into fruit shelf life extension via manipulation of cuticle content.

Formulating Electrolytes for 4.6 V Anode-Free Lithium Metal Batteries

High-voltage initial anode-free lithium metal batteries (AFLMBs) promise the maximized energy densities of rechargeable lithium batteries. However, the reversibility of the high-voltage cathode and lithium metal anode is unsatisfactory in sustaining their long lifespan. In this research, a concentrated electrolyte comprising dual salts of LiTFSI and LiDFOB dissolved in mixing solvents of dimethyl carbonate (DMC) and fluoroethylene carbonate (FEC) with a LiNO3 additive was formulated to address this challenge. FEC and LiNO3 regulate the anion-rich solvation structure and help form a LiF, Li3N-rich solid electrolyte interphase (SEI) with a high lithium plating/stripping Coulombic efficiency of 98.3%. LiDFOB preferentially decomposes to effectively suppress the side reaction at the high-voltage operation of the Li-rich Li1.2Mn0.54Ni0.13Co0.13O2 cathode. Moreover, the large irreversible capacity during the initial charge/discharge cycle of the cathode provides supplementary lithium sources for cycle life extension. Owing to these merits, the as-fabricated AFLMBs can operate stably for 80 cycles even at an ultrahigh voltage of 4.6 V. This study sheds new insights on the formulation of advanced electrolytes for highly reversible high-voltage cathodes and lithium metal anodes and could facilitate the practical application of AFLMBs.

Natural variation in age-related dopamine neuron degeneration is glutathione dependent and linked to life span

Aging is the biggest risk factor for Parkinson's disease (PD), suggesting that age-related changes in the brain promote dopamine neuron vulnerability. It is unclear, however, whether aging alone is sufficient to cause significant dopamine neuron loss, and if so, how this intersects with PD-related neurodegeneration. Here, through examining a large collection of naturally varying Drosophila strains, we find a strong relationship between life span and age-related dopamine neuron loss. Strains with naturally short-lived animals exhibit a loss of dopamine neurons without generalized neurodegeneration, while animals from long-lived strains retain dopamine neurons across age. Metabolomic profiling reveals lower glutathione levels in short-lived strains which is associated with elevated levels of reactive oxygen species (ROS), sensitivity to oxidative stress, and vulnerability to silencing the familial PD gene parkin. Strikingly, boosting neuronal glutathione levels via glutamate-cysteine ligase (Gcl) overexpression is sufficient to normalize ROS levels, extend life span, and block dopamine neurons loss in short-lived backgrounds, demonstrating that glutathione deficiencies are central to neurodegenerative phenotypes associated with short longevity. These findings may be relevant to human PD pathogenesis, where glutathione depletion is reported to occur in the idiopathic PD patient brain through unknown mechanisms. Building on this, we find reduced expression of the Gcl catalytic subunit in both Drosophila strains vulnerable to age-related dopamine neuron loss and in the human brain from familial PD patients harboring the common LRRK2 G2019S mutation. Our study across Drosophila and human PD systems suggests that glutathione synthesis and levels play a conserved role in regulating age-related dopamine neuron health.

Dietary restriction impacts health and lifespan of genetically diverse mice.

Caloric restriction extends healthy lifespan in multiple species1. Intermittent fasting, an alternative form of dietary restriction, is potentially more sustainable in humans, but its effectiveness remains largely unexplored2-8. Identifying the most efficacious forms of dietary restriction is key for developing interventions to improve human health and longevity9. Here we performed an extensive assessment of graded levels of caloric restriction (20% and 40%) and intermittent fasting (1 and 2 days fasting per week) on the health and survival of 960 genetically diverse female mice. We show that caloric restriction and intermittent fasting both resulted in lifespan extension in proportion to the degree of restriction. Lifespan was heritable and genetics had a larger influence on lifespan than dietary restriction. The strongest trait associations with lifespan included retention of body weight through periods of handling-an indicator of stress resilience, high lymphocyte proportion, low red blood cell distribution width and high adiposity in late life. Health effects differed between interventions and exhibited inconsistent relationships with lifespan extension. 40% caloric restriction had the strongest lifespan extension effect but led to a loss of lean mass and changes in the immune repertoire that could confer susceptibility to infections. Intermittent fasting did not extend the lifespan of mice with high pre-intervention body weight, and two-day intermittent fasting was associated with disruption of erythroid cell populations. Metabolic responses to dietary restriction, including reduced adiposity and lower fasting glucose, were not associated with increased lifespan, suggesting that dietary restriction does more than just counteract the negative effects of obesity. Our findings indicate that improving health and extending lifespan are not synonymous and raise questions about which end points are the most relevant for evaluating aging interventions in preclinical models and clinical trials.