Senescence Markers Research Articles

Changes in senescence markers after a weight loss intervention in older adults with obesity

BackgroundUnderstanding how weight loss interventions in older adults with obesity impact aging biology can lay the foundation for targeted, ‘geroscience-based’ interventions. This study examines the association between changes in the senescence-associated secretory phenotypes (SASP) and changes in function in response to a weight loss intervention. MethodsWe conducted a post-hoc biomarker analysis on adults aged ≥ 65 years with body mass index [BMI] ≥30 kg/m2 enrolled in a six-month, non-randomized telemedicine-delivered weight loss intervention. We assessed 16 SASP cytokines using serum samples collected pre-and post-intervention. Clinical outcomes include anthropometric and physical function measurements. A weight loss responder was defined as a loss of ≥5 % of body weight. ResultsMean age was 73.2 ± 3.9 years (73 % female), and BMI was 36.5 ± 5.2 kg/m2. Responders lost 7.6 ± 2.5 %, while non-responders lost 2.0 ± 2.3 % of weight (n = 16 per group, p < 0.001). We observed several significant associations between SASP cytokines and physical function and anthropometric measurement outcomes in age- and sex-adjusted linear models. These included grip strength and Interleukin-8 (IL-8) (b = 9.07) and Insulin-like Growth Factor 1 (IGF-1) (b = 2.6); gait speed and Thymus and Activation-Regulated Chemokine (TARC) (b = 0.46) and IL-7(b = 61 0.11); weight IL-6 (b = -6.77) and IL-15 (b = -2.53); BMI and IL-15 (b = -0.95); waist-to-hip ratio and osteopontin (b = -0.07) (p < 0.05 for all). ConclusionsOur pilot data demonstrated an association between changes in select SASP biomarkers and increased functional ability with intentional weight loss in older adults with obesity. However, findings must be replicated in prospective randomized trials with a control group and additional SASP biomarkers.

Physical activity and DNA methylation-based markers of ageing in 6208 middle-aged and older Australians: cross-sectional and longitudinal analyses.

Epigenetic age quantifies biological age using DNA methylation information and is a potential pathway by which physical activity benefits general health. We aimed to assess the cross-sectional and longitudinal associations between physical activity and epigenetic age in middle-aged and older Australians. Blood DNA methylation data for 6208 participants (40% female) in the Melbourne Collaborative Cohort Study (MCCS) were available at baseline (1990-1994, mean age, 59years) and, of those, for 1009 at follow-up (2003-2007, mean age, 69years). Physical activity measurements (weighted scores at baseline and follow-up and total MET hours per week at follow-up) were calculated from self-reported questionnaire data. Five blood methylation-based markers of ageing (PCGrimAge, PCPhenoAge, bAge, DNAmFitAge, and DunedinPACE) and four fitness-related markers (DNAmGaitspeed, DNAmGripmax, DNAmVO2max, and DNAmFEV1) were calculated and adjusted for age. Linear regression was used to examine the cross-sectional and longitudinal associations between physical activity and epigenetic age. Effect modification by age, sex, and BMI was assessed. At baseline, a standard deviation (SD) increment in physical activity was associated with 0.03-SD (DNAmFitAge, 95%CI = 0.01, 0.06, P = 0.02) to 0.07-SD (bAge, 95%CI = 0.04, 0.09, P = 2 × 10-8) lower epigenetic age. These associations were attenuated after adjustment for other lifestyle variables. Only weak evidence was found for the longitudinal association (N = 1009) of changes in physical activity and epigenetic age (e.g. DNAmFitAge: adjusted β = - 0.04, 95%CI = - 0.08, 0.01). The associations were not modified by age, sex, or BMI. In middle-aged and older Australians, higher levels of self-reported physical activity were associated with slightly lower epigenetic age.

FXR1 associates with and degrades PDZK1IP1 and ATOH8 mRNAs and promotes esophageal cancer progression

BackgroundThe growing body of evidence suggests that RNA-binding proteins (RBPs) have an important function in cancer biology. This research characterizes the expression status of fragile X-related protein 1 (FXR1) in esophageal cancer (ESCA) cell lines and understands its mechanistic importance in ESCA tumor biology.MethodsThe role of FXR1, PDZK1IP1, and ATOH8 in the malignant biological behaviors of ESCA cells was investigated using in-vitro and in-vivo experiments.ResultsFXR1 was aberrantly overexpressed at both the transcript and protein levels in ESCA cells. Deficiency of FXR1 in ESCA cells was associated with decreased cell proliferation, viability and compromised cell migration compared to the control group. In addition, the inhibition of FXR1 leads to the promotion of apoptosis and cell cycle arrest in ESCA cells. Furthermore, FXR1 knockdown stabilizes senescence markers, promoting cellular senescence and decreasing cancer growth. Mechanistically, FXR1 negatively regulated PDZK1IP1 or ATOH8 transcripts by promoting mRNA degradation via direct interaction with its 3′UTR. PDZK1IP1 or ATOH8 overexpression predominantly inhibited the tumor-promotive phenotype in FXR1-overexpressed cells. Furthermore, FXR1 inhibition and PDZK1IP1 or ATOH8 overexpression in combination with FXR1-overexpressed cells significantly decreased xenograft tumor formation and enhanced nude mouse survival without causing apparent toxicity (P < 0.01). In the FXR1 knockdown group, the tumor weight of mice decreased by 80% compared to the control group (p < 0.01).ConclusionsOur results demonstrate FXR1’s oncogenic involvement in ESCA cell lines, suggesting that FXR1 may be implicated in ESCA development by regulating the stability of PDZK1IP1 and ATOH8 mRNAs. For the first time, our findings emphasize the importance of FXR1-PDZK1IP1 and -ATOH8 functional modules in the development of ESCA, which might have potential diagnostic or therapeutic implications.

Marker gene fishing for single-cell data with complex heterogeneity.

In single-cell studies, cells can be characterized with multiple sources of heterogeneity such as cell type, developmental stage, cell cycle phase, activation state, and so on. In some studies, many nuisance sources of heterogeneity (SOH) are of no interest, but may confound the identification of the SOH of interest, and thus affect the accurate annotate the corresponding cell subpopulations. In this paper, we develop B-Lightning, a novel and robust method designed to identify marker genes and cell subpopulations correponding to a SOH (e.g., cell activation status), isolating it from other sources of heterogeneity (e.g., cell type, cell cycle phase). B-Lightning uses an iterative approach to enrich a small set of trustworthy marker genes to more reliable marker genes and boost the signals of the SOH of interest. Multiple numerical and experimental studies showed that B-Lightning outperforms existing methods in terms of sensitivity and robustness in identifying marker genes. Moreover, it increases the power to differentiate cell subpopulations of interest from other heterogeneous cohorts. B-Lightning successfully identified new senescence markers in ciliated cells from human idiopathic pulmonary fibrosis (IPF) lung tissues, new T cell memory and effector markers in the context of SARS-COV-2 infections, and their synchronized patterns which were previously neglected. This paper highlights B-Lightning's potential as a powerful tool for single-cell data analysis, particularly in complex data sets where sources of heterogeneity of interest are entangled with numerous nuisance factors.

Antisenescence Expansion of Mesenchymal Stem Cells Using Piezoelectric β-Poly(vinylidene fluoride) Film-Based Culture.

Regenerative therapies based on mesenchymal stem cells (MSCs) show promise in treating a wide range of disorders. However, the replicative senescence of MSCs during in vitro expansion poses a challenge to obtaining a substantial quantity of high-quality MSCs. In this investigation, a piezoelectric β-poly(vinylidene fluoride) film-based culture plate (β-CP) was developed with an antisenescence effect on cultured human umbilical cord-derived MSCs. Compared to traditional tissue culture plates (TCPs) and α-poly(vinylidene fluoride) film-based culture plates, the senescence markers of p21, p53, interleukin-6 and insulin-like growth factor-binding protein-7, stemness markers of OCT4 and NANOG, and telomere length of MSCs cultured on β-CPs were significantly improved. Additionally, MSCs at passage 18 cultured on β-CPs showed significantly better multipotency and pro-angiogenic capacities in vitro, and higher wound healing abilities in a mouse model. Mechanistically, β-CPs rejuvenated senescent MSCs by improving mitochondrial functions and mitigating oxidative and glycoxidative stresses. Overall, this study presents β-CPs as a promising approach for efficient and straightforward antisenescence expansion of MSCs while preserving their stemness, thereby holding great potential for large-scale production of MSCs for clinical application in cell therapies.

Multiomic profiling reveals timing of menopause predicts prefrontal cortex aging and cognitive function.

A new case of dementia is diagnosed every 3 s. Beyond age, risk prediction of dementia is challenging. There is growing evidence of underlying processes that connect aging across organ systems and may provide insight for early detection, and there is a need to identify early biomarkers at an age when action can be taken to mitigate cognitive decline. We hypothesized that timing of menopause, a marker of ovarian aging, predicts brain age decades later. We used 2086 subjects with multiple "omics" measurements from post-mortem brain samples. Age at menopause (AAM) is positively correlated with cognitive function and negatively correlated with pre-frontal cortex aging acceleration (calculated as estimated biological age from DNA methylation minus chronological age). Genetic correlations showed that at least part of these associations is derived from shared heritability. To dissect the mechanism linking AAM to cognitive decline, we turned to transcriptomic data which confirmed that later AAM was associated with gene expression in pre-frontal cortex consistent with better cognition, and among those who reached menopause naturally, decreased gene expression of pathways implicated in aging. Those with surgical menopause displayed different molecular changes, including perturbed nicotinamide adenine dinucleotide (NAD+) activity, validated by metabolomics. Bile acid metabolism was perturbed in both groups, although different bile acid ratios were associated with AAM in each. Together, our data suggest that AAM is predictive of brain aging and cognition, with potential mediation by the gut, although through different mechanisms depending on the type of menopause.

Modifiable dementia risk associated with smaller white matter volume and altered 1/f aperiodic brain activity: cross-sectional insights from the LEISURE study.

The rising prevalence of dementia necessitates identifying early neurobiological markers of dementia risk. Reduced cerebral white matter volume and flattening of the slope of the electrophysiological 1/f spectral power distribution provide neurobiological markers of brain ageing alongside cognitive decline. However, their association with modifiable dementia risk remains to be understood. A cross-sectional sample of 98 healthy older adults (79 females, mean age = 65.44) underwent structural magnetic resonance imaging (sMRI), resting-state electroencephalography (EEG), cognitive assessments and dementia risk scoring using the CogDrisk framework. Univariate and multivariate linear regression models were conducted to investigate the relationships between modifiable dementia risk and sMRI brain volumes, the exponent of EEG 1/f spectral power, and cognition, whilst controlling for non-modifiable factors. Smaller global white matter volume (F(1,87) = 6.884, R2 = 0.073, P = .010), and not grey (F(1,87) = 0.540, R2 = 0.006, P = .468) or ventricle volume (F(1,87) = 0.087, R2 = 0.001, P = .769), was associated with higher modifiable dementia risk. A lower exponent, reflecting a flatter 1/f spectral power distribution, was associated with higher dementia risk at frontal (F(1,92) = 4.096, R2 = 0.043, P = .046) but not temporal regions. No significant associations were found between cognitive performance and dementia risk. In multivariate analyses, both white matter volume and the exponent of the 1/f spectral power distribution independently associated with dementia risk. Structural and functional neurobiological markers of early brain ageing, but not cognitive function, are independently associated with modifiable dementia risk in healthy older adults.

Metabolite signatures of chronological age, aging, survival, and longevity

Metabolites that mark aging are not fully known. We analyze 408 plasma metabolites in Long Life Family Study participants to characterize markers of age, aging, extreme longevity, and mortality. We identify 308 metabolites associated with age, 258 metabolites that change over time, 230 metabolites associated with extreme longevity, and 152 metabolites associated with mortality risk. We replicate many associations in independent studies. By summarizing the results into 19 signatures, we differentiate between metabolites that may mark aging-associated compensatory mechanisms from metabolites that mark cumulative damage of aging and from metabolites that characterize extreme longevity. We generate and validate a metabolomic clock that predicts biological age. Network analysis of the age-associated metabolites reveals a critical role of essential fatty acids to connect lipids with other metabolic processes. These results characterize many metabolites involved in aging and point to nutrition as a source of intervention for healthy aging therapeutics.

Modeling of TDP-43 proteinopathy by chronic oxidative stress identifies rapamycin as beneficial in ALS patient-derived 2D and 3D iPSC models

Amyotrophic Lateral Sclerosis (ALS) is a fatal neurodegenerative disorder characterized neuropathologically by TDP-43 proteinopathy with loss of TDP-43 nuclear splicing activity and formation of cytoplasmic TDP-43 aggregates. The lack of suitable experimental models of TDP-43 proteinopathy has hampered the discovery of effective therapies. We already showed that chronic and mild oxidative insult by sodium arsenite (ARS) triggered TDP-43 cytoplasmic aggregation and stress granules (SGs) formation in ALS patient-derived fibroblasts and motor neurons differentiated from induced pluripotent stem cells (iPSC-MNs). However, whether this insult induces a reduction of TDP-43 splicing activity in the nucleus, thus recapitulating both gain and loss of function pathomechanisms, still remains to be determined.In this study we first showed that chronic ARS in human neuroblastoma cells triggered TDP-43 cytoplasmic mislocalization, SGs formation and defective splicing of TDP-43 target genes UNC13A and POLDIP3 as functional readouts of TDP-43 proteinopathy. Additionally, a dysregulation of autophagy and senescence markers was observed in this condition. In a preliminary drug screening approach with autophagy-promoting drugs, namely rapamycin, lithium carbonate and metformin, only rapamycin prevented ARS-induced loss of TDP-43 splicing activity. We then demonstrated that, in addition to TDP-43 cytoplasmic aggregation, chronic ARS triggered TDP-43 loss of splicing activity also in ALS patient-derived primary fibroblasts and iPSC-MNs and that rapamycin was beneficial to reduce these TDP-43 pathological features. By switching to a neuro-glial 3D in vitro model, we observed that treatment of ALS iPSC-brain organoids with chronic ARS also induced a defective TDP-43 splicing activity which was prevented by rapamycin.Collectively, we established different human cell models of TDP-43 proteinopathy which recapitulate TDP-43 gain and loss of function, prevented by rapamycin administration. Human neuroblastoma cells and patient-derived fibroblasts and 2D- and 3D-iPSC models exposed to chronic oxidative stress represent therefore suitable in vitro platforms for future drug screening approaches in ALS.

Ionizing radiation has negligible effects on the age, telomere length and corticosterone levels of Chornobyl tree frogs.

The accident that occurred at the Chornobyl nuclear power plant (Ukraine, 1986) contaminated a large extension of territory after the deposition of radioactive material. It is still under debate whether the chronic exposure to the radiation levels currently present in the area has long-term effects on organisms, such as decreases in longevity. Here, we investigate whether current levels of radiation in Chornobyl negatively impact the age of the Eastern tree frog Hyla orientalis. We also explore whether radiation induces changes in an ageing marker, telomere length or the stress hormone corticosterone. We found no effect of total individual absorbed radiation (including both external and internal exposure) on frog age (n = 197 individuals sampled in 3 consecutive years). We also did not find any relationship between individual absorbed radiation and telomere length, nor between individual absorbed radiation and corticosterone levels. Our results suggest that radiation levels currently experienced by Chornobyl tree frogs may not be high enough to cause severe chronic damage to semi-aquatic vertebrates such as this species. This is the first study addressing age and stress hormones in Chornobyl wildlife, and thus future research will confirm if these results can be extended to other taxa.

Association Between TyG Index, Liver Steatosis and Immunosenescence in People Living with HIV.

Metabolic disorders and immunosenescence increase the risk of complications in people living with HIV (PLWH), affecting mortality and quality of life. However, their relationship remains unclear. Participants were grouped by median TyG index, and logistic regression identified baseline independent factors of a high TyG index at Week 24. The association of the TyG index for hepatic steatosis was determined using ROC curves. We also explored correlations between the TyG index and aging markers, including CD4/CD8 ratio and CD8+ T cells and evaluated health-related quality of life (HRQoL). A total of 203 PLWH were included in the study. We observed that PLWH in high TyG group tended to be older (P<0.001), have greater body weight (P<0.001), higher ALT levels (P=0.021), and increased low-density lipoprotein levels (P=0.001). ROC analysis revealed that TyG index was closely associated with hepatic steatosis at Week 52 (AUC=0.743) and Week 104 (AUC=0.728). Moreover, a higher TyG index was positively correlated with CD8+ T cell counts, while patients in the high TyG group had lower CD4/CD8 ratios at Week 52 and Week 104. Poorer mental health was observed in patients with CD8+ T cell counts ≥1000 and a high TyG index. Multivariate analysis further identified baseline older age (OR=1.108, P=0.002), elevated cholesterol (OR=3.407, P<0.001), and low HDL (OR=0.003, P<0.001) as factors associated with a high TyG index at Week 24. The TyG index is closely linked to metabolic disorders and immunosenescence in PLWH. It offers a basis for personalized treatment strategies, improving physical and mental health and reducing complication risks.

Comparative analysis of various senescence inducers in proximal renal tubular cells

Senescence in renal cells has attracted wide attention as the critical factor promoting renal fibrosis and chronic kidney disease. Establishing a reliable cellular model is essential to study mechanisms underlying renal cell senescence. Herein, we compared various inducers to define the most suitable senescence inducer for HK-2 proximal tubular cells. These inducers included hydrogen peroxide (H2O2), high-temperature (HT), glucose, mannitol and hydroxyurea (HU). To screen for optimal concentration/level, the highest concentration/level of each inducer that did not increase cell death (to avoid severe toxicity) was selected for senescence induction and comparative analysis using the two most appropriate markers for HK-2 cell senescence as recently established. The data revealed that 0.4 mM, 43 °C, 80 mM, 80 mM and 100 μM were the optimal concentrations/levels of H2O2, HT, glucose, mannitol and HU, respectively. Comparative analysis using optimal concentration/level of each marker revealed that 0.4 mM H2O2, HT at 43 °C, 80 mM glucose and 80 mM mannitol were the weak senescence inducers. The most effective inducer for HK-2 senescence was 100 μM HU, which provided the greatest fold-changes of cell area and granularity when compared with other stimuli in a time-dependent manner. Based on these data comparing H2O2, HT, glucose, mannitol and HU at their optimal concentrations/levels, 100 μM HU seems to be most effective for senescence induction in HK-2 cells for in vitro study of proximal renal tubular cells.

Matrix Gla protein suppresses osteoblast senescence and promotes osteogenic differentiation by the PI3K-AKT signaling pathway

Age-related bone loss in mice is associated with senescent cell accumulation and reduced bone formation by osteoblasts. Matrix Gla protein (MGP), secreted by osteoblasts, is pivotal in regulating the bone extracellular matrix mineralization. Previous research has demonstrated that Mgp null mice exhibit osteopenia and fractures, and ultimately die prematurely. To elucidate the mechanisms underlying MGP’s role of MGP in bone metabolism, we generated osteoblast-specific Mgp knockout (Mgp cKO) mice by crossing Mgpfl/fl mice with Bglap-Cre mice. The study revealed that in 3-month-old Mgp cKO male mice, trabecular bone volume decreased, and the senescence marker protein p21 increased. Primary osteoblasts from Mgp cKO mice exhibited markers of DNA damage and senescence, such as increased γH2AX foci, p21, and senescence-associated β-galactosidase staining, as well as attenuated cellular proliferation and osteogenic differentiation abilities. In addition, bone marrow stromal cells’ colony formation and spontaneous osteogenic ability were impaired in Mgp cKO mice, whereas osteoclastogenesis was enhanced. In vitro treatment with recombinant human MGP promotes osteogenesis in osteoblasts derived from Mgp cKO mice via the PI3K-AKT signaling pathway. Thus, our results suggest that MGP is protective by suppressing osteoblast senescence, offering new insights into potential therapeutic strategies for age-related osteoporosis.

The association between hair cortisol and burnout is moderated by age, psychosocial, and immunological markers

BackgroundExhaustion and depersonalization are the core symptoms of the occupational burnout. However, burnout is not an all-or-nothing phenomenon, but can occur in a milder to moderate form in otherwise healthy employees. In the last two decades hair cortisol concentrations (HCC) were increasingly related to the cumulative effect of psychosocial stress at work. We analyzed data of the Dortmund Vital Study (Clinicaltrials.gov: NCT05155397) to explore the relationship of HCC and burnout symptoms. Moreover, we asked whether the HCC – burnout association was moderated by work ability, chronic stress, neuroticism, depressive symptoms, and stress-related immunological biomarkers such as T cell concentration, CD4/CD8 cell ratio, and proinflammatory cytokines TNF- α, IL-6, and IL-18. MethodsBurnout was assessed by the Oldenburg Burnout Inventory (OLBI), and the Maslach Burnout Inventory (MBI) in 196 working adults aged between 20 and 65 years (mean age 42.2 years). Several self-reported variables and biomarkers were collected. ResultsThe results showed an association between HCC and the burnout measures. A series of moderator analyses revealed that the association between HCC and burnout symptoms was substantial for low work ability, high chronic stress level, high neuroticism level, and mild to moderate depressive symptoms. Immunological markers moderated the HCC – burnout association for high concentrations of T cells, low CD4/CD8 ratio and low IL-6, IL-18 and TNF-α concentrations. These interactions were moderated by age showing the largest impact in middle-aged to older individuals. ConclusionsThe present findings shed light on the complex interaction between burnout symptoms and work ability, chronic stress, personality, and the endocrinological and immunological responses across the working lifespan. These parameters should be considered when assessing the risk for developing burnout and validating the diagnosis of burnout. Trial RegistrationClinicalTrials.gov NCT05155397; https://clinicaltrials.gov/ct2/show/NCT05155397

Dawn-to-dusk intermittent fasting is associated with overexpression of autophagy genes: A prospective study on overweight and obese cohort

Aim and BackgroundA growing body of evidence supports the impact of intermittent fasting (IF) on longevity and healthy aging via the modulation of autophagy genes. The activation of the catabolic autophagic machinery (LAMP2, LC3B, ATG5, and ATG4D) has protective effects against degenerative aging and chronic diseases. This research examined the changes in the expression of the aforementioned genes upon the observance of dawn-to-dusk IF among metabolically healthy participants with overweight and obesity. MethodsFifty-one (51) participants (36 males and 15 females, 38.84 ± 11.73 years) with overweight and obesity (BMI = 29.75 ± 5.04 kg/m2) were recruited and monitored before and at the end of the commencement of the four-week IF. Six healthy subjects with normal BMI (21.4±2.20 kg/m2) were recruited only to standardize the reference for normal levels of gene expressions. At the two time points, anthropometric, biochemical, and dietary assessments were performed, and LAMP2, LC3B, ATG5, and ATG4D gene expressions were assessed using qRT-PCR on RNA extracted from whole blood samples. ResultsAt the end of IF, and compared to the pre-fasting levels, the relative gene expressions among participants with overweight/obesity were significantly increased for the three autophagy genes LAMP2, LC3B, and ATG5, with increments of about 4.2 folds, 1.9-fold, and 1.4-fold, respectively. In contrast, the increase in the ATG4D gene was not significant. Concomitantly, significant decreases were found in body weight, BMI, fat mass, body fat percent, hip and waist circumferences, LDL, IL-6, and TNF-a (P <0.05), While HDL, IL-10, and CD163 significantly increased (P <0.05). Binary logistic regression analysis for genetic expressions showed no significant association between high-energy intake, waist circumference, or obesity and the four gene expressions. ConclusionsFour consecutive weeks of dawn-to-dusk IF of Ramadan is associated with the upregulation of autophagy gene expressions in participants with overweight/obesity, and this may explain, at least in part, its favorable short-term temporal metabolic and health-improving effects on early aging-related markers. Hence, IF presumably may entail a protective impact against early markers of aging and metabolic diseases in participants with overweight/obesity.

P53 dependence of senescence markers p21v1 and p21v2 in aging and acute injury

p53 dependence of senescence markers p21v1 and p21v2 in aging and acute injury

Histomorphometric changes in pituitary gonadotropic endocrinocytes when exposed to dark deprivation

Aim. To assess the effect of 30-day dark deprivation on functional and histomorphometric changes in adenohypophysis gonadotropic endocrinocytes and their reversibility in mature male rats.Materials and methods. Mongrel white male rats (n = 36) weighing 365–375 g at 4 months of age were randomly divided into three groups (each n = 12). For 30 days the control group was in automatic light-dark mode 12/12, and the rats of experimental groups 1 and 2 were in round-the-clock artificial lighting (24/0, 300 Lux), then the rats of group 2 were returned to 12/12 mode for the next 14 days. In the animals of the control and group 1 during their lifetime on the 31st day, and in group 2 on the 45th day, blood was taken from the abdominal aorta and levels of follicle-stimulating (FSH) and luteinizing (LH) hormones, melatonin, and Klotho protein were determined an enzyme-linked immunosorbent assay and immunoassay and after which they were removed from the experiment by decapitation. Postmortem histological and immunohistochemical examination of the pituitary gland was done using rabbit polyclonal antibodies targeting caspase-3 and Klotho protein, as well as morphometry. Statistical data processing was performed using the Kruskal-Wallis test with post-hoc Dunn’s test.Results. Light desynchronization in the form of 30 days of dark deprivation increased FSH and LH levels and decreased melatonin and Klotho protein levels in the blood of male rats; increased gonadotropic endocrine cell area, volume, and perimeter by 23.1% (p &lt; 0.001), 48.7% (p &lt; 0.001), and 10.9% (p &lt; 0.001), respectively; and increased nucleus area, volume, and perimeter by 16%, 11.7%, and 2.5%, respectively. An immunohistochemical study showed an increase in the specific area of caspase-3-immunoreactive gonadotropic endocrinocytes by 25.2% without obvious morphological signs of apoptosis, and a decrease in the expression of Klotho protein by 25.7%. All indicators were reversible, the levels of FSH and Klotho protein in the blood of animals almost reached their initial values after 14 days of restoration of the light-dark cycle 12/12.Conclusion. Dark deprivation for 30 days in male rats induced reversible processes of accelerated aging and apoptosis in cells, as evidenced by changes in the expression of aging markers in gonadotropic endocrinocytes and levels of gonadotropic hormones in the blood. When the light-dark mode is restored, the levels of FSH and Klotho protein normalize as early as 14 days.

Telomere length across the spectrum of metabolic health – an analysis from the LIPIDOGEN2015 study

IntroductionBackground: Telomere length is a cellular aging marker and correlates with various cardiovascular disease (CVD) risk factors. The current study assessed the association between obesity, metabolic syndrome (MetS), and telomere length.Material and methodsMaterial and methods: The LIPIDOGRAM&amp;LIPIDOGEN2015 study was conducted in primary care in 2015-2016. Recruited patients to the LIPIDOGEN2015 cohort (n=1788) were a random subset of patients of the LIPIDOGRAM2015 (n=13,724) study. For the aims of this analysis, the recruited patients were divided into four groups based on the presence of MetS: healthy slim (HS), metabolically healthy obese (MHO), non-obese with MetS (NOMS), and metabolically unhealthy obese (MUO). Relative telomere length (RTL) was measured using quantitative polymerase chain reaction (qPCR).ResultsResults: 1516 patients (85%; females - 59.7%, mean age- 50.3 years) were included for final analyses. An increase in body mass index (BMI), waist circumference, prevalence of diabetes mellitus, hypertension, dyslipidemia, and history of myocardial infarction moving from HS to MUO were observed. MUO group exhibited the highest triglycerides and lowest high-density lipoprotein (HDL-C) levels. Univariable regression analyses indicated that NOMS (p=0.038) and MUO (p=0.003) were associated with significantly decreased RTL. After adjustment for age, gender, education, smoking, place of residence, and myocardial infarction, the association was no longer statistically significant.ConclusionsConclusions: Despite the lack of statistical significance in the multivariate analysis, the univariate results suggest that both MUO and NOMS phenotypes contribute to the shortening of telomere length. These results may also indicate that MetS, irrespectively on obesity occurrence, is responsible for the shortened lifespan.

Considerations on Multimorbidity and Frailty in Inflammatory Bowel Diseases.

There are growing numbers of older people with inflammatory bowel diseases [IBD]. These older patients are more likely to have other comorbidities and polypharmacy, which can make recognizing and treating IBD complex. Frailty is a newer concept in the IBD field, and we are beginning to recognize the importance of this as a marker of biological age and its association with risk of adverse IBD-related outcomes. In this review article we aim to provide practical insight into the specific challenges facing older patients and their clinicians at each stage of the patient journey. We also discuss the latest understanding of the impact of frailty for these patients with IBD and highlight areas for future research.

Single Cell MALDI-MSI Analysis of Lipids and Proteins within a Replicative Senescence Fibroblast Model.

In this study, we evaluate lipids and select proteins in human lung fibroblasts (hLFs) to interrogate changes occurring due to aging and senescence. To study single cell populations, a comparison of cells adhered onto slides using poly-d-lysine versus centrifugal force deposition was first analyzed to determine whether specific alterations were observed between preparations. The poly-d-lysine approach was then utilized to interrogate the lipidome of the cell populations and further evaluate potential applications of the MALDI-immunohistochemistry (IHC) platform for single-cell-level analyses. Two protein markers of senescence, vimentin and p21, were both observed within the fibroblast populations and quantified. Lipidomic analysis of the fibroblasts found 12 lipids significantly altered because of replicative senescence, including fatty acids, such as stearic acid, and ceramide phosphoethanolamine species (CerPE). Similar to previous reports, alterations were detected in putative fatty acid building blocks, ceramides, among other lipid species. Altogether, our results reveal the ability to detect lipids implicated in senescence and show alterations to protein expression between normal and senescent fibroblast populations, including differences between young and aged cells. This report is the first time that the MALDI-IHC system has been utilized at a single-cell level to analyze both protein expression and lipid profiles in cultured cells, with a particular focus on changes associated with aging and senescence.