ABSTRACT Epstein–Barr virus (EBV) is a lifelong persistent herpesvirus, usually asymptomatic but associated with several malignancies and autoimmune disorders. EBV encodes viral proteins and microRNAs (miRNAs) that modulate host gene expression and facilitate immune evasion. This pilot study analyzed host cellular miR- NA expression in EBV-positive (EBV+) and EBV-negative (EBV-) individuals, as well as within EBV infection subgroups, and examined IL-6 promoter methylation and telomere length. In a cohort of 156 participants, EBV infection was confirmed in 95 individuals via qPCR and categorized as chronic latent, chronic lytic, acute early, or acute recent. miR-328 and miR-21 significantly differentiated EBV+ from EBV- individuals, while miR-877 distin- guished acute from chronic infections. IL-6 promoter methylation inversely correlated with miR-328 expression, indicating an association between epigenetic regulation of in- flammation in EBV persistence. No telomere length differences were observed between EBV+ and EBV- groups. These findings are hypothesis-generating and support further mechanistic work on miRNA–epigenetic interactions in EBV persistence.

IntroductionThe extension of human longevity has intensified the search for biomarkers that capture not only chronological age but also biological aging and functional healthspan. Among molecular candidates, microRNAs (miRNAs) have emerged as promising regulators and indicators of aging-related processes. In this pilot study, we explored whether selected circulating miRNAs could serve as potential biomarkers of biological age and lifestyle-associated aging dynamics.MethodsBased on current literature, we focused on three miRNAs—miR-24, miR-21, and miR-155—previously linked to inflammation, senescence, and metabolic regulation. Capillary blood samples from a heterogeneous adult cohort were analyzed using quantitative PCR. ΔCt values were integrated into a composite “miRNA-3Age” model through multivariate regression analysis to estimate biological age. Associations between lifestyle variables (diet, exercise, stress, and smoking) and miRNA-based biological age were examined.ResultsThe miRNA-3Age model predicted biological age with moderate correlation to chronological age and revealed variability consistent with individual health profiles. Participants with favorable lifestyle factors (e.g., frequent consumption of fish, whole grains, and green tea; regular exercise) tended to exhibit lower miRNA-3Age estimates, whereas stress and smoking were associated with higher predicted biological age.DiscussionOur exploratory data suggest that integrating multiple miRNA signals may enhance the sensitivity of biological age estimation compared to single-biomarker approaches. However, variability within the small sample highlights the need for larger, longitudinal datasets to confirm predictive validity and to disentangle causal links between lifestyle, miRNA expression, and aging biology.ConclusionThis pilot study supports the feasibility of miRNA-based biological age modeling and identifies miR-24, miR-21, and miR-155 as promising components of a composite biomarker framework. The miRNA-3Age model provides a preliminary step toward a scalable, lifestyle-sensitive aging metric that warrants validation in diverse populations.

Introduction:Polyphenols are a group of compounds identified as secondary metabolites of plants, with 8,000 types identified to date. Previous research findings have indicated the potential anti-inflammatory properties of polyphenols, with studies suggesting a reduction in disease risk and therapeutic benefits observed in various diseases, including diabesity, neurodegeneration, cancer, and cardiovascular disease. Objective:The objective of this study was to comprehensively analyze the polyphenol composition of extracts of Greek mountain tea (GMT) and wheat germ (WG) and investigate their effects on microcirculation and eicosanoid metabolism. Materials and methods:The polyphenol and spermidine composition of GMT and WGE was analyzed using LC-HRMS. Hemodynamic impact of GMT or WG on rat cremasteric arteriole blood flow was measured after compound administration using a laser Doppler blood flow meter. Lipidomic analysis in urine after co-administration of GMT and WGE was measured by LC-HRMS mass spectrometry. Results:This study shows that GMT contains large amount of polyphenols, expecially ferulic acid and petunidin. In contrast, in the WG extract we found minimal polyphenol content. Subsequent to the administration of GMT to rats, a significant increase in rat cremasteric arteriole blood flow was observed, while WG extract exhibited minimal change. Following a single oral administration of GMT or WG to mice, 24 h urine was analyzed for eicosanoids. A significant decrease in pro-inflammatory eicosanoids and a substantial increase in anti-inflammatory eicosanoids were observed in the treatment group compared with the control group. Conclusions:Given the established role of polyphenol intake in enhancing vascular endothelial function and increasing peripheral blood flow, we suggest that the observed increase in blood flow is a consequence of polyphenols in GMT. In contrast, the enhancement of eicosanoid balance was more pronounced in the WG extract group compared to the GMT group, suggesting that this effect may be attributable to components other than polyphenols present in these fractions.

Obesity is considered to be a chronic complex disease that increases the risk of diabetes, heart diseases and certain cancers. According to the literature, in obese people the gastrointestinal microbiota is disturbed, which could be the cause of the onset of obesity and related diseases. Moreover, there are small non-coding RNAs (microRNAs) that are disturbed in obesity, which are also considered to be a possible mechanism of probiotics' action. Certain microRNAs are involved in the development and metabolism of adipose tissue cells, as well as the secretion and action of insulin, showing that changed expression of certain miRNAs could have a significant impact on the onset and development of obesity and obesity-related diseases. Thus, miRNAs are considered to be possible markers for the diagnosis and prognosis of various metabolic diseases, and possible therapeutic targets for the treatment of obesity and related diseases. Considering the growing need of the healthcare system for nutraceuticals and dietary supplements that present effective and safe medical nutritive therapy in obese individuals, this work aimed to assess the beneficial effects of probiotics on obesity-related microRNAs. In this review, we described the role of microRNAs and probiotics in obesity and the association between probiotics and obesityrelated microRNAs. Circulating miRNAs' profile in obese individuals significantly differs from that of normal-weight individuals. miRNAs such as miR-155, miR-221, miR-24-3p, and miR181a are over-expressed, while miR-26b and 125a are under-expressed in obese patients compared to non-obese individuals. It has been found that supplementation with Bifidobacterium bifidum and Lactobacillus acidophilus could downregulate the expression of miR-155 and miR221, and upregulate the expression of miR-26b. Supplementation with a probiotic formulation containing S. boulardii, L. plantarum 299v, and octacosanol led to the downregulation of miR155 and miR-24-3p expression. miR-125a and miR-181a were upregulated and downregulated, respectively, after the intervention with L. delbrueckii and L. rhamnosus. After reviewing the available literature, we can conclude that probiotics have beneficial effects on microRNAs altered in obesity, which could provide an effective strategy for the management of obesity.

The importance of diet and lifestyle in maintaining overall health has long been recognised. MicroRNAs (miRNAs) have emerged as key players in the intricate interplay between health and disease. This study, including 305 participants, examined the role of miRNAs from capillary blood as indicators of individual physiological characteristics, diet, and lifestyle influences. Key findings include specific miRNAs associated with inflammatory processes and dietary patterns. Notably, miR-155 was associated with subjects with metabolic diseases and upregulated in age. Additionally, the study revealed diet-related miRNA expressions: high consumption of vegetables, fruits, and whole grains correlated with increased levels of miR-let-7a and miR-328, both implicated in anti-inflammatory pathways, and decreased expression of pro-inflammatory miR-21. In the context of smoking, we found a significant decrease in miRNA-142, known for its downregulation in lung cancer. We observed a sex-biased expression of various miRNAs with significant upregulation of miR-151a in females and a higher expression of miR-155 in ageing females, representing a possible mechanism for the increased susceptibility to autoimmune diseases. In conclusion, the study underscores the significant influence of lifestyle, nutrition, and sex on miRNA profiles. Circulating miRNAs demonstrate significant potential as biomarkers in personalized medicine, highlighting their utility in tailoring healthcare to individual needs.

Dietary spermidine is emerging as a substance with promising health-promoting and healthy ageing-promoting properties. Its autophagy-inducing potential has been especially well-documented. This process is suspected to be at the root of the life-prolonging effects of fasting and in-vivo spermidine supplementation. In humans, high dietary spermidine intake is associated with a lifespan increase of 5.7 years. Numerous studies provide indications of its effects on the cardiovascular, nervous, and immune systems as well as on the formation and development of tumors. Nevertheless, no health claims are currently authorized in the EU for spermidine or spermidine-rich functional foods. When ingested through food or dietary supplements, dosage appears to play an important role in replicating the favorable effects. Currently, there are ongoing proceedings regarding the novelty of a certain manufacturing process of spermidine supplements, but many other plant extracts with high spermidine content from different sources are marketed without the need for novel food authorization. This article discusses the varying test results obtained for the spermidine concentration of popular spermidine supplements. Soybean extract seems to be slightly superior in terms of spermidine concentration, but one wheat germ-based product stood out in this analysis due to a 10 times higher spermidine concentration than comparable products.Keywords: spermidine, food supplements, novel food, botanicals, functional foods, autophagy, spermidine-rich wheat germ extract

Dysregulation of epigenetic mechanisms has been recognized to play a crucial role in cancer development, but these mechanisms vary between sexes. Therefore, we focused on sex-specific differences in the context of cancer-based data from a recent study. A total of 12 cell-free DNA methylation targets in CpG-rich promoter regions and 48 miRNAs were analyzed by qPCR in plasma samples from 8 female and 7 male healthy controls as well as 48 female and 80 male subjects with solid tumors of the bladder, brain, colorectal region (CRC), lung, stomach, pancreas, and liver. Due to the small sample size in some groups and/or the non-balanced distribution of men and women, sex-specific differences were evaluated statistically only in healthy subjects, CRC, stomach or pancreas cancer patients, and all cancer subjects combined (n female/male—8/7, 14/14, 8/15, 6/6, 48/80, respectively). Several miRNAs with opposing expressions between the sexes were observed for healthy subjects (miR-17-5p, miR-26b-5p); CRC patients (miR-186-5p, miR-22-3p, miR-22-5p, miR-25-3p, miR-92a-3p, miR-16-5p); stomach cancer patients (miR-133a-3p, miR-22-5p); and all cancer patients combined (miR-126-3p, miR-21-5p, miR-92a-3p, miR-183-5p). Moreover, sex-specific correlations that were dependent on cancer stage were observed in women (miR-27a-3p) and men (miR-17-5p, miR-20a-5p). Our results indicate the complex and distinct role of epigenetic regulation, particularly miRNAs, depending not only on the health status but also on the sex of the patient. The same miRNAs could have diverse effects in different tissues and opposing effects between the biological sexes, which should be considered in biomarker research.

Background Psychological stress, as an important cofactor in the development of many acute and chronic diseases, is crucial for general health or well-being, and improved markers are needed to distinguish situations of progressive pathological development, such as depression, anxiety, or burnout, to be recognized at an early stage. Epigenetic biomarkers play an important role in the early detection and treatment of complex diseases such as cancer, and metabolic or mental disorders. Therefore, this study aimed to identify so-called miRNAs, which would be suitable as stress-related biomarkers. Methods and Results In this study, 173 participants (36.4% males, and 63.6% females) were interviewed about stress, stress-related diseases, lifestyle, and diet to assess their acute and chronic psychological stress status. Using qPCR analysis, 13 different miRNAs (miR-10a-5p, miR-15a-5p, miR-16-5p, miR-19b-3p, miR-26b-5p, miR-29c-3p, miR-106b-5p, miR-126-3p, miR-142-3p, let-7a-5p, let-7g-5p, miR-21-5p, and miR-877-5p) were analyzed in dried capillary blood samples. Four miRNAs were identified, miR-10a-5p, miR-15a-5p, let-7a-5p, and let-7g-5p ( p < 0.05), which could be used as possible candidates for measuring pathological forms of acute or chronic stress. Let-7a-5p, let-7g-5p, and miR-15a-5p ( p < 0.05) were also significantly higher in subjects with at least one stress-related disease. Further, correlations were identified between let-7a-5p and meat consumption ( p < 0.05) and between miR-15a-5p and coffee consumption ( p < 0.05). Conclusion The examination of these four miRNAs as biomarkers using a minimally invasive method offers the possibility of detecting health problems at an early stage and counteracting them to maintain general and mental health.

Background: Viruses significantly affect global health. The few available viricidal and antiviral therapies are expensive and often associated with unwanted side effects. Functional foods as plant extracts (nutraceuticals) are epigenetically active, and multi-target antiviral compounds that affect several steps of the viral life cycle and host proteins. MicroRNAs (miRNAs) post-transcriptionally regulate host and viral gene expression and are responsible for the fine-tuning of gene expression by controlling the expression of their target messenger RNAs (mRNA) in host cells and viruses. Several plant ingredients have been proven to be active against RNA virus infections. Obviously, miRNAs play a central role in the regulation of gene transcription in viral replication and host immune defense. A healthy diet and nutraceuticals can support the fine-tuning of miRNAs. Objectives: The aim of this study was to analyze the effects of epigenetic active phytoceuticals and/or functional foods on immune relevant miRNAs and mRNAs in a healthy human study population. Methods: In a 2-month human intervention study (n=80), a mix of plant extracts (sage extract, green tea extract, berberine, apple extract, onion extract, elderberry extract, and grape skin extract) with proven activity against RNA viruses were examined for the change of miRNA and mRNA patterns involved in virus response systems. Lifestyle and nutritional behavior, including self-reported stress levels, as well as infection frequency as potential cofounders have been raised. Results: The intervention with secondary plant extracts could modulate the expression of miRNAs involved in the virus defense (miR-30e, miR-877, miR-150) and the expression of NF-κB. The intervention also increased miRNA expression patterns associated with a positive lifestyle. The intervention adapted miRNAs that correlate with an increased risk of infection, smoking, stress, and processed meat products.Conclusion: Plant extracts can enhance the beneficial effects of a healthy nutrition and lifestyle by modulating miRNAs. MiRNAs are promising drug targets for maintaining immune homeostasis.Keywords: antiviral, epigenetic active nutraceutical, miRNA, secondary plant extracts, virus response, plant ingredients, functional foods

Healthy mitochondria and their epigenetic control are essential to maintaining health, extending life expectancy, and improving cardiovascular performance. Strategies to maintain functional mitochondria during aging include training; cardiovascular exercise has been suggested as the best method, but strength training has also been identified as essential to health and healthy aging. We therefore investigated the effects of concurrent exercise training and dietary habits on epigenetic mechanisms involved in mitochondrial (mt) functions and biogenesis. We analyzed epigenetic biomarkers that directly target the key regulator of mitochondrial biogenesis, PGC-1α, and mtDNA content. Thirty-six healthy, sedentary participants completed a 12-week concurrent training program. Before and after the intervention, dried blood spot samples and data on eating habits, lifestyle, and body composition were collected. MiR-23a, miR-30e expression, and mtDNA content were analyzed using real-time quantitative polymerase chain reaction (qPCR) analysis. PGC-1α methylation was analyzed using bisulfite pyrosequencing. MiR-23a, miR-30e expression, and PGC-1α methylation decreased after the intervention (p < 0.05). PGC-1α methylation increased with the consumption of red and processed meat, and mtDNA content increased with the ingestion of cruciferous vegetables (p < 0.05). Our results indicate that concurrent training could improve mitochondrial biogenesis and functions by altering the epigenetic regulation. These alterations can also be detected outside of the skeletal muscle and could potentially affect athletic performance.

Liquid biopsy-based tests emerge progressively as an important tool for cancer diagnostics and management. Currently, researchers focus on a single biomarker type and one tumor entity. This study aimed to create a multi-analyte liquid biopsy test for the simultaneous detection of several solid cancers. For this purpose, we analyzed cell-free DNA (cfDNA) mutations and methylation, as well as circulating miRNAs (miRNAs) in plasma samples from 97 patients with cancer (20 bladder, 9 brain, 30 breast, 28 colorectal, 29 lung, 19 ovarian, 12 pancreas, 27 prostate, 23 stomach) and 15 healthy controls via real-time qPCR. Androgen receptor p.H875Y mutation (AR) was detected for the first time in bladder, lung, stomach, ovarian, brain, and pancreas cancer, all together in 51.3% of all cancer samples and in none of the healthy controls. A discriminant function model, comprising cfDNA mutations (COSM10758, COSM18561), cfDNA methylation markers (MLH1, MDR1, GATA5, SFN) and miRNAs (miR-17-5p, miR-20a-5p, miR-21-5p, miR-26a-5p, miR-27a-3p, miR-29c-3p, miR-92a-3p, miR-101-3p, miR-133a-3p, miR-148b-3p, miR-155-5p, miR-195-5p) could further classify healthy and tumor samples with 95.4% accuracy, 97.9% sensitivity, 80% specificity. This multi-analyte liquid biopsy-based test may help improve the simultaneous detection of several cancer types and underlines the importance of combining genetic and epigenetic biomarkers.

Background Regular, especially sustained exercise plays an important role in the prevention and treatment of multiple chronic diseases. Some of the underlying molecular and cellular mechanisms behind the adaptive response to physical activity are still unclear, but recent findings suggest a possible role of epigenetic mechanisms, especially miRNAs, in the progression and management of exercise-related changes. Due to the combination of the analysis of epigenetic biomarkers (miRNAs), the intake of food and supplements, and genetic dispositions, a "fitness score" was evaluated to assess the individual response to nutrition, exercise, and metabolic influence. Methods In response to a 12-week sports intervention, we analyzed genetic and epigenetic biomarkers in capillary blood from 61 sedentary, healthy participants (66.1% females, 33.9% males, mean age 33 years), including Line-1 methylation, three SNPs, and ten miRNAs using HRM and qPCR analysis. These biomarkers were also analyzed in a healthy, age- and sex-matched control group (n, 20) without intervention. Food frequency intake, including dietary supplement intake, and general health questionnaires were surveyed under the supervision of trained staff. Results Exercise training decreased the expression of miR-20a-5p, -22-5p, and -505-3p (p < 0.02) and improved the "fitness score," which estimates eight different lifestyle factors to assess, nutrition, inflammation, cardiovascular fitness, injury risk, regeneration, muscle and hydration status, as well as stress level. In addition, we were able to determine correlations between individual miRNAs, miR-20a-5p, -22-5p, and -101-3p (p < 0.04), and the genetic predisposition for endurance and/or strength and obesity risk (ACE, ACTN3, and FTO), as well as between miRNAs and the body composition (p < 0.05). MiR-19b-3p and -101-3p correlated with the intake of B vitamins. Further, miR-19b-3p correlated with magnesium and miR-378a-3p with iron intake (p < 0.05). Conclusions In summary, our results indicate that a combined analysis of several biomarkers (miRNAs) can provide information about an individual's training adaptions/fitness, body composition, nutritional needs, and possible recovery. In contrast to most studies using muscle biopsies, we were able to show that these biomarkers can also be measured using a minimally invasive method.

Background: Sirtuins attract high attention considering their properties to reverse molecular hallmarks of aging and age-related disorders. Many secondary plant ingredients (SPI) are known for their sirtuin-activating activities as well as epigenetic regulation of telomers, autophagy, senolysis, DNA repair but also improvement of gut microbiota. Furthermore, prebiotics enhanced butyrate was shown to interact with SIRT pathways. This study investigated effects of a drink containing a mix of different SPIs in combination with galactooligosaccharides (GOS) and their effect on SIRT activation, markers of aging relevant mechanisms and gut microbiota composition in correlation with subjective wellbeing and skin structure appearance. Methods: We analyzed gene expression, mtDNA amount, and microbial composition in response to a sirtuin-activating drink in humans compared to a control group consuming a placebo. Food frequency, beauty, and general health questionnaires were asked, and a set of mRNAs and miRNAs were assessed using qPCR analysis. The gut composition was analyzed using Illumnia sequencing.Results: SPI increased SIRT1, SIRT3 and modulated cell cycle relevant miR16 and senescence regulating miR34 expression. Additionally, mtDNA amount was higher in the group consuming the active supplement indicating an improved mitochondrial activity. The combined effect of SPI and GOS lead to an increase of Actinobacteria, especially Bifidobacterium, but also Veillonellaceae which was not observed in the control group. Significant correlations between SIRT3 expression and the gut microbiota Bifidobacterium and Veillonellaceae were observed. Additionally, statistical analysis of subjects self-reporting indicated beneficial effects regarding beauty and wellbeing.Conclusion: Our results show that the combination of sirtuins inducing SPI and prebiotic GOS influences molecular pathways counteracting aging, senescence, inflammation, and enhanced groups of gut microbiota which are known to improve the innate and adaptive immune system. Keywords: secondary plant ingredients, prebiotic, Sirtuins, subjective wellbeing, Bifidobacterium

Introduction: Topical investigations have demonstrated that oxidative stress and inflammation play key roles in biological aging and determine incidence and course of age-related diseases. Lifestyle and environmental factors hugely impact epigenetic regulation and DNA stability with telomere attrition and epigenetic instability providing a potential record of the cumulative burden of endogenous and exogenous oxidative noxae. Certain physiologically active plant components exhibit antioxidative activities affecting epigenetic regulation of inflammation response and DNA repair.Methods: Against this background, the present study investigated green tea polyphenol epigallocatechin gallate (EGCG) in the context of telomere regulation in Caco-2 colorectal adenocarcinoma cells vs. ES-1 primary skin fibroblasts. Cell lines were treated with 20 and 200 µM EGCG for 36, 72 and 144 hours, respectively. Telomerase activity, relative telomere length as well as methylation status of hTERT and c-Myc from different culture conditions were assessed. Malondialdehyde (MDA) served as a surrogate marker of potential pro-oxidative effects of EGCG in a physiologically relevant tissue model.Results: EGCG incubation was associated with telomere shortening and decreased telomerase activity in Caco-2 cells, and relatively longer telomeres along with increased methylation of six 5'—C—phosphate—G—3' (CpG) sites in the promoter region of human Telomerase Reverse Transcriptase (hTERT) in fibroblasts. At low concentrations, EGCG significantly decreased oxidative damage to lipids in Caco-2 cells and attenuated H2O2 induced oxidation at higher concentrations.Conclusion: These results suggest differential EGCG-mediated telomeric modulation in cancer vs. primary cells and a specific antioxidant activity of EGCG against oxidative damage to lipids in abnormal cells.Keywords: Caco-2, epigallocatechin gallate, telomeres, hTERT, DNA methylation, telomerase, oxidative stress, malondialdehyde

Periodic fasting (PF) is an increasingly popular approach that assists in the management of metabolic and inflammatory diseases as well as in preventing mechanisms involved in aging. However, little is known about the effects of fasting on gut microbiota and its impact on the epigenetic regulation of metabolically relevant enzymes, especially sirtuins (SIRTs). We analyzed the effect of periodic fasting on the human gut microbiota, SIRTs expression, and mitochondrial content in 51 males and females. The participants fasted under supervision for five consecutive days following the Buchinger fasting guidelines. Ketogenesis, selected mRNAs, miRNAs, mitochondrial (mt) DNA, and gut composition were analyzed before and after PF. PF triggered a significant switch in metabolism, as indicated by the increase in ß-hydroxybutyrate (BHB) and pyruvate dehydrogenase kinase isoform 4 (PDK4) expression in the capillary blood. MtDNA, SIRT1, SIRT3, and miRlet7b-5p expression in blood cells were elevated, whereas SIRT6 and miR125b-5p were not affected. Following fasting, gut microbiota diversity increased, and a statistically significant correlation between SIRT1 gene expression and the abundance of Prevotella and Lactobacillus was detected. The abundance of longevity related Christensenella species increased after fasting and inversely correlated with age as well as body mass index (BMI). Thus, this represents the first study that showing that fasting not only changes the composition of the gut microbiota, making it more diverse, but also affects SIRT expression in humans.

Following research obtained from the previous SARS and MERS outbreaks, we’ve gained knowledge about the mechanisms of bioactive plant ingredients against the attachment and replication of COVID-19 as well as overshooting immune responses. This could be used for designing COVID-19 trials utilizing bioactive compounds. The receptors for SARS, ACE-2, and CD26 show associations with mechanisms that regulate human senescence. Several functional foods interact with the epigenetic regulation of viral infection and mechanisms of senescence. This review concentrates on the link between bioactive plant ingredients and their activities against mechanisms of viral infections. Keywords: COVID-19, Epigenetic, Quercetin, Curcumin, Epigallocatechin gallate, Phloretin, Berberine

Background: Fasting and fasting mimetics - bioactive compounds mimicking fasting effects, are of growing interest as potential means to slow down the aging process and increase health span. Sirtuins are known as enzymes that interfere with mitochondrial energy metabolism and molecular pathways involved in longevity. Although their activation is determined as a response to stress i.e. caloric restriction. Sirtuin activating nutraceuticals are believed to mimic the effects of nutrient deprivation, thus activating signaling pathways correlated to an improved health span. In this study, we compare 5 days periodic buchinger fasting intervention with 3 months shot supplementation, a drink formula, containing secondary plant ingredients considered to activate sirtuins.Methods: We analyzed pathways in response to fasting and a sirtuins activating drink. Genetic and epigenetic biomarkers including telomere length, LINE1 methylation, and a set of mRNAs and miRNAs were assessed using qPCR analysis. Gut composition and metabolites were compared using Illumnia sequencing and mass spectrometry.Results: Fasting, but also the fasting mimetic could increase expression of FoxO1, SIRT1, and MLH1 mRNA, all genes discussed in aspects of longevity. A positive correlation between telomere length and both SIRT1, and SIRT6 was observed. Furthermore, a significant change in the gut composition was measured. Actinobacteria increased in the supplementation group, whereas after buchinger fasting a rise in the distribution of Proteobacteria could be observed. Firmicutes/Bacteroidetes ratio decreased and correlated with the body mass index (BMI).Conclusions: Our results confirm the effects of fasting on longevity associated mechanisms but also suggest that SIRTFOOD shot intervention addresses some of these effects.

AIM:We investigated different bioactive compounds including epigallocatechin gallate (EGCG), anthocyanidin, resveratrol, phloretin, spermidine, butyrate, and β-hydroxybutyrate with regard to their effect on SIRT3 via NRF2 and modulation of the proinflammatory senescence-associated secretory phenotype (SASP) in senescence induced 3T3-L1 preadipocytes.METHODS:For induction of senescence, 3T3-L1 preadipocytes were incubated with bromodeoxyuridine (BrdU) for 8 days. Cell cycle inhibition was observed, and β-galactosidase activity was measured. After BrdU treatment, cells were treated with different bioactive compounds in various concentrations for 96 h. ELISA was used for determining proinflammatory cytokine IL6 in SASP cells.RESULTS:CDKN1a increased significantly after BrdU incubation compared to untreated control (p < 0.01). All secondary plant ingredients used for treatment, but not anthocyanidin 50 μM, decrease CDKN1a expression (p < 0.05), whereas most endogenous substances did not attenuate CDKN1a. IL6 secretion positively correlated with CDKN1a (p < 0.01), whereas EGCG could diminish both, IL6 and CDKN1a with the strongest effect (p < 0.01). Although NRF2 positively correlated with SIRT3 activation (p < 0.05), only resveratrol (p < 0.01) and anthocyanidin (p < 0.05) could activate NRF2 significantly. Solely anthocyanidin 50 μM (p < 0.05) and 100 μM (p < 0.01) and EGCG 50 μM (p < 0.01) could increase SIRT3 expression. Activation of SIRT3 with EGCG correlated with lowered IL6 secretion significantly (p < 0.05) but not with anthocyanidin.CONCLUSION:Accumulation of senescent cells in adipose tissue plays an important role in obesity and age-related diseases. SIRT3, located in the mitochondria, can regulate ROS via different pathways. Thus, targeting SIRT3 activating compounds such as EGCG may delay senescence of cells and senescence induced inflammatory processes.