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BACKGROUND: Changes in DNA methylation along the life have been documented, and environmental exposures (including diet, physical activity and smoking) can accelerate or decelerate this process. The epigenetic clock estimates the biological age of an individual measuring methylation patterns in specific areas of its genome. Recently, a new epigenetic clock based on 6 CpGs has been proposed, with high potential to become an easy accessible tool able to measure the epigenetic age (EA) of an individual. OBJECTIVES: This study aims to validate the 6 CpG epigenetic clock comparing it with other biomarkers of aging such as telomere length (TL) and methylation in the long interspersed nuclear element-1 (LINE-1). The impact of lifestyle associated factors on these molecular marks has been evaluated. METHODS: 200 healthy participants having extreme dietary patterns (healthy vs western diet) were selected. Dietary intakes, body composition, physical activity level and smoking has been assessed. DNA extracted from whole blood was used to measure the 6CpG-EA, TL and LINE-1 methylation levels. RESULTS: 6CpG-EA was positively correlated with chronological age (r = 0.591; p = 7.2*10-20) and negatively with TL (r = –0.150; p = 0.040) and LINE-1 methylation (r = –0.240; p = 0.001). Despite the fact that no significant associations were detected with the overall diet quality (HEI), 6CpG-EA was correlated with dietary intakes of nutrients involved in the one-carbon (1 C) metabolism, especially in the western diet group. CONCLUSION: These results support the 6CpG epigenetic clock as an easy accessible tool to estimate biological age, in accordance with other molecular markers of aging, and suggest that EA can be modulated by micronutrients involved in the 1 C metabolism.

Purpose Some dietary habits cluster together, and for this reason it is advised to study the impact of entire dietary patterns on human health, rather than that of individual dietary habits. The main objective of this study was to evaluate differences in gut microbiota composition and their predicted functional properties between people with a healthy (HDP) and western (WDP) dietary pattern. Methods A cross-sectional, observational study was carried out on 200 participants enrolled 2017–2018 in Poznań, Poland, equally distributed into HDP and WDP groups. Diet was estimated using 3-day food records and information on stool transit times was collected. Fecal microbiota composition was assessed by 16S rRNA gene sequencing and its functional properties were predicted by the PICRUSt2 workflow. Results The α-diversity did not differ between people with WDP and HDP, but β-diversity was associated with dietary pattern. People with HDP had higher relative abundances (RA) of Firmicutes and Faecalibacterium and lower RA of Bacteroidota and Escherichia–Shigella than participants with WDP. Only a small proportion of the variance in microbiota composition (1.8%) and its functional properties (2.9%) could be explained by dietary intake (legumes, simple sugars and their sources, like fruit, soft drinks) and stool transit characteristics. Conclusion Gut microbiota composition and predicted metabolic potential is shaped by overall diet quality as well as the frequency of defecation; however, the cumulative effect of these explain only a relatively low proportion of variance.

Coffee is one of the most consumed beverages in the world, but the extent to which it is consumed is affected by both environmental and genetic factors. Genome-wide association studies and candidate date association studies have identified several gene variants associated with increased consumption of coffee. Functional single-nucleotide polymorphisms in rs762551 (cytochrome P450 1A2 [CYP1A2]) and rs5751876 (adenosine receptor A2A [ADORA2A]) has been linked to individual caffeine response. Coffee intake has been shown to affect lipid metabolism. We thus hypothesize that rs762551 (CYP1A2) A allele carriers consume more coffee than C allele carriers and that rs5751876 (ADORA2A) C allele carriers consume less coffee than T allele carriers. Additionally, we hypothesize that CYP1A2 genotype can modulate serum glucose concentrations and lipid profile. A total of 421 participants aged 20 to 40 years were recruited from 2016 to 2018 in Poznań, Poland. Genotyping of CYP1A2 and ADORA2A was performed using TaqMan probes. Individuals with AA CYP1A2 genotype consumed relatively more coffee with milk (72.81 ± 10.15 mL/1000 kcal vs 43.38 ± 6.42 mL/1000 kcal, P = .008) and with milk or cream than did C allele carriers, whereas the rs5751876 ADORA2A polymorphism was not associated with coffee or tea intake. Additionally, subjects with AA CYP1A2 genotype had 10% higher serum triacylglycerol (TG) concentrations than C allele carriers. This study suggests that CYP1A2 rs762551 polymorphism is associated with coffee intake and serum TG concentrations in healthy 20- to 40-year-old adults.

Several previous investigations have employed betaine supplementation in randomized controlled crossover designs to assess its ostensible ergogenic potential. Nevertheless, prior methodology is predicated on limited pharmacokinetic data and an appropriate betaine-specific washout period is hitherto undescribed. The purpose of the present pilot investigation was therein to determine whether a 28 day washout period was sufficient to return serum betaine concentrations to baseline following a supplementation protocol. Five resistance-trained men (26 ± 6 y) supplemented with 6 g/day betaine anhydrous for 14 days and subsequently visited the lab 10 additional times during a 28 day washout period. Participants underwent venipuncture to assess serum betaine and several other parameters before (PRE) and periodically throughout the washout timeframe (POST0, -4, -7, -10, -13, -16, -19, -22, -25 and -28). All analyses were performed at a significance level of p < 0.05. While analyses failed to detect any differences in any other serum biomarker (p > 0.05), serum betaine was significantly elevated from PRE-to-POST0 (p = 0.047; 2.31 ± 1.05 to 11.1 ± 4.91 µg·mL−1) and was statistically indistinguishable from baseline at POST4 (p = 1.00). Nevertheless, visual data assessment and an inability to assess skeletal muscle concentrations would otherwise suggest that a more conservative 7 day washout period is sufficient to truly return both serum-and-skeletal muscle betaine content to pre-supplementation levels.

Although some bacteria inhabiting the human gut synthesize folates, it has not yet been established whether bacterial folate biosynthesis can impact human folate status. The main objectives of this study were to evaluate associations between different lifestyle factors and the potential of fecal microbiota to produce folates, and to investigate whether this potential is associated with circulating folate and total homocysteine (tHcy) levels in humans. To this end, we carried out an observational study of two hundred adult participants, with high variance in dietary habits. Diet was determined using three-day food records. Fecal microbiota composition was assessed by 16S rRNA gene amplicon sequencing. To establish the folate-production potential of fecal bacteria, cultures containing feces were incubated under anaerobic conditions for 24 h, and the folate concentration was measured before and after incubation. The folate concentration in cultures was 185.4 ± 228.1 pg/ml/log(CFU/g) (2125.4 ± 2454.3 pg/ml) higher after incubation. This change in concentration was not associated with the healthy eating index that measures diet quality (r = -0.11, p = 0.11), but it was positively associated with low α-diversity (r = -0.18, p < 0.01), and high relative abundance of the Bacteroides, as well as Sutterella and Parasutterella genera. The gut microbiota's folate producing potential was associated neither with serum folate nor with plasma tHcy levels. In conclusion, some taxa of the native gut microbiota have the ability to synthesize folates under culture conditions, but this bacterial folate biosynthesis capacity does not predict human folate status.

Mitochondrial DNA copy number (mtDNAcn) has been proposed for use as a surrogate biomarker of mitochondrial health, and evidence suggests that mtDNA might be methylated. Intermediates of the one-carbon cycle (1CC), which is duplicated in the cytoplasm and mitochondria, have a major role in modulating the impact of diet on the epigenome. Moreover, epigenetic pathways and the redox system are linked by the metabolism of glutathione (GSH). In a cohort of 101 normal-weight and 97 overweight/obese subjects, we evaluated mtDNAcn and methylation levels in both mitochondrial and nuclear areas to test the association of these marks with body weight, metabolic profile, and availability of 1CC intermediates associated with diet. Body composition was associated with 1CC intermediate availability. Reduced levels of GSH were measured in the overweight/obese group (p=1.3∗10−5). A high BMI was associated with lower LINE-1 (p=0.004) and nominally lower methylenetetrahydrofolate reductase (MTHFR) gene methylation (p=0.047). mtDNAcn was lower in overweight/obese subjects (p=0.004) and independently correlated with MTHFR methylation levels (p=0.005) but not to LINE-1 methylation levels (p=0.086). DNA methylation has been detected in the light strand but not in the heavy strand of the mtDNA. Although mtDNA methylation in the light strand did not differ between overweight/obese and normal-weight subjects, it was nominally correlated with homocysteine levels (p=0.035) and MTHFR methylation (p=0.033). This evidence suggests that increased body weight might perturb mitochondrial-nuclear homeostasis affecting the availability of nutrients acting as intermediates of the one-carbon cycle.

“Western diet” containing high concentrations of sugar and fat consumed during pregnancy contributes to development of obesity and diabetes type 2 in offspring. To mimic effects of this diet in animals, a cafeteria (CAF) diet is used. We hypothesized that CAF diet given to rats before, and during pregnancy and lactation differently influences fat content, metabolic and inflammation profiles in offspring. Females were exposed to CAF or control diets before pregnancy, during pregnancy and lactation. At postnatal day 25 (PND 25), body composition, fat contents were measured, and blood was collected for assessment of metabolic and inflammation profiles. We have found that CAF diet lead to sex-specific alterations in offspring. At PND25, CAF offspring had: (1) higher percentage of fat content, and were lighter; (2) sex-specific differences in levels of glucose; (3) higher levels of interleukin 6 (IL-6), interleukin-10 (IL-10) and tumor necrosis factor (TNF-α); (4) sex-specific differences in concentration of IL-6 and TNF-α, with an increase in CAF females; (5) higher level of IL-10 in both sexes, with a more pronounced increase in females. We concluded that maternal CAF diet affects fat content, metabolic profiles, and inflammation parameters in offspring. Above effects are sex-specific, with female offspring being more susceptible to the diet.

The present study aimed at analysing how dietary folic acid (FA) and Fe deficiency, followed by supplementation with these nutrients, affects the expression of folate and Fe transporters in the duodenum, as well as FA and Fe status. After a deficiency period, Wistar rats were randomised to a group fed with a diet deficient in FA and supplemented with Fe (DFE), a diet deficient in Fe and supplemented with FA, a diet supplemented with Fe and FA (FEFOL), a diet deficient in Fe and FA (D) or a control diet (C). Tissue collection was performed after 2, 10 or 21 d of these diets. Group D had higher Slc11a2 mRNA levels than the DFE group at every time point and there were differences in mRNA levels of Slc46a1 between the DFE and the FEFOL groups at the third time point, but we observed no differences in protein levels between the groups. The DFE and D groups not only had lower serum folate concentrations at every time point but also had the highest homocysteine concentrations. Total Fe binding capacity concentrations were the lowest in the DFE group at the first time point and in the DFE and the FEFOL groups at the final time point. Simultaneous supplementation with FA and Fe resulted in significantly higher Hb concentrations than did supplementation with these nutrients alone. Our findings indicate that dietary FA and Fe deficiency, and subsequent supplementation with these nutrients, affects transcription but not the protein levels of FA and Fe transporters in the duodenum.

Although simultaneous supplementation with iron and folic acid is justified, the potential interactions between these micronutrients are unknown. The aim of this study was to determine the effects of oral iron and folic acid, administered together or separately, on iron concentration in tissues in rats with a deficiency of both these micronutrients. In the first stage of the experiment (28 days), 150 8-week-old female Wistar rats were randomly assigned to a control group (C; n = 30) fed the standard diet and to a study group (n = 120) fed a diet deficit in iron and folate. The study group was then randomly divided to four groups: D group fed a deficit diet, FE group fed a deficit diet with iron gluconate, the FOL group fed a deficit diet with folate acid, and the FEFOL group fed a deficit diet with iron gluconate and folate acid. After 2, 10, and 21 days of supplementation, ten animals from each group were killed. Morphological parameters were measured in whole blood. Iron concentration was assayed in serum, liver, spleen, pancreas, heart, and kidneys. Folic acid supplementation more significantly decreased iron concentrations in the pancreas and spleen than in the D group after 10 and 21 days of supplementation. Moreover, the combination of iron with folic acid markedly decreased iron levels in the liver and spleen, in comparison with iron alone, after 10 and 21 days of the experiment. In conclusion, folic acid affects iron status in female rats deficient in these micronutrients in moderate and long-term supplementation.

BACKGROUND Little is known about the relation between iron and folic acid (FA) supplementation and inflammation. The aim of this study was to evaluate the effects of iron and folate deficiency and supplementation on blood morphology parameters, and to assess the role of iron and folate transporters in inflammation. METHODS A four-week period of FA and iron deficiency in Wistar rats was followed by randomization into a group fed with a diet deficient in FA and supplemented with Fe (DFE), a group fed a diet deficient in Fe and supplemented with FA (DFOL), a group fed a diet supplemented with Fe and FA (FEFOL), a group fed a diet deficient in Fe and FA (D), and a group fed a control diet (C). The blood Crp concentration and blood count were determined. The expression of SLC11A2, SLC46A1, SLC19A1, and TFR2 proteins was assessed using the western blot method. RESULTS After ten days on the experimental diets, the rats in the DFOL group had a 21% higher concentration of white blood cells (WBC) than the FEFOL group did (p < 0.05). We did not observe any differences between the groups in terms of C-reactive protein (Crp) concentration. We also did not find any other differences between the groups in other morphological parameters. Analysis of the correlation between blood count parameters and the expression of iron and folate transporters gave conflicting results. CONCLUSIONS To conclude, iron and folate supplementation may affect WBC concentration in the blood.

Although simultaneous supplementation of iron and folic acid is justified, the potential interactions between these micronutrients and other elements are poorly known. In this study, we aimed to investigate the effect of iron and folic acid supplementation on the levels of selected essential and toxic elements in the serum of micronutrient-deficient young women. A total of 40 women participated in this study and were divided into two groups: study group (n = 23) (with iron and folate deficiency) and control group (n = 17). The study group received iron and folic acid supplements for 3 months. Blood samples were collected at baseline and after the completion of the study period. Women completed a 3-day food intake record. We calculated the body mass index (BMI) of all the participants. Cellular morphology was analyzed in whole blood, and biochemical parameters were determined in serum. Elements were measured in serum by inductively coupled plasma mass spectrometry (ICP-MS). According to our results, in the case of the study group, the supplementation of iron and folic acid restored their levels, however, it caused a significant decrease in the level of zinc, calcium, and magnesium. In the case of the control group, at the end of the study period, there was a marked decrease in the level of iron. Interestingly, there was an increase in the level of arsenic and vanadium in both groups. In conclusion, simultaneous supplementation of iron and folic acid impairs the level of zinc, calcium, and magnesium in women of childbearing age.

Syftet med denna uppsats är att ta reda på vilka socioekonomiska faktorer bidrar till intaget av vaccinet mot covid-19 i 13 olika bostadsområden i Stockholm. Författarna av denna uppsats åtar sig undersöka fyra samband med vaccinationsgraden i de utvalda områden, det vill säga samband med utbildning, inkomst, arbetslöshet och utländsk bakgrund. Tidigare forskning i frågan har påvisat motstridiga resultat, dock formas hypoteserna utifrån resultatet som återfinns i majoriteten studier. För att berika diskussionen tas även upp begrepp som institutionell tillit, infodemic och vaccintveksamhet. Metoden består av linjära regressionsmodeller där de oberoende variablerna i form av socioekonomiska faktorer redovisas och konstanthålls i flera kombinationer. Variablerna redovisas först för sig i enkla linjära regressioner, där inkomst och hög utbildning korrelerar positivt med vaccinationsgraden och arbetslöshet och utländsk bakgrund korrelerar negativt med vaccinationsgraden. Vidare görs regressionsmodeller om två och tre oberoende variabler. Överraskande i denna studie är att i detta skede visar sig inkomst ha ett negativt eller icke-signifikant samband med vaccinationsgraden. Även utbildning förlorar sin signifikans till vaccinationsgraden. Däremot behåller utländsk bakgrund sin signifikanta negativa korrelation med vaccinationsgraden. Avslutningsvis diskuteras variablerna i förhållande till den tidigare forskningen. The purpose of this study is to find out which socioeconomic factors contribute to the intake of the covid-19 vaccine in 13 different residential areas in Stockholm. The authors of the following thesis undertake to research the association of four socioeconomic factors with the degree of vaccination in said areas, that is education level, average income, unemployment and immigrant background. Previous research in the question has shown contradictory evidence, the hypothesis of this study however is formulated in line with the majority of the research. To further enrich the discussion on the topic, notions of institutional trust, infodemic and vaccine hesitancy are considered. Additionally, tests of linear regression models are performed and adjusted in several combinations. Firstly, the independent variables are presented by themselves, where education and income correlate positively with the vaccination degree, whereas unemployment and immigrant background correlate negatively. Furthermore, linear regressions with two and three independent variables are performed. The surprising result here is that income either no longer has a significant correlation or correlates negatively with vaccination degree. Education level also loses its significance. On the other hand, immigrant background withholds its negative correlation with the vaccination degree. Finally, the results are discussed in relation to the prior research.

Nonalcoholic fatty liver disease (NAFLD) is widely prevalent globally and has no effective treatment. Coffee is one of the most popular beverages in the world and can therefore have a significant impact on public health on account of its health-promoting properties. Evidence from observational, clinical, and animal studies suggests that coffee may play an important role in human health. This article summarizes the effects of coffee on liver health, especially on nonalcoholic fatty liver disease (NAFLD) and its progression: liver fibrosis, cirrhosis and hepatocellular carcinoma. In addition, this article describes the pathogenesis, prevalence, diagnosis, and nutrition guidelines relating to NAFLD. Possible mechanisms responsible for the effects of coffee on the liver are also suggested.

Supplementation with iron and folic acid is widely recommended in women of childbearing age and during pregnancy; however, the effect of such supplementation on mineral status is not well-known. The aim of this study was to determine the effects of oral iron and folic acid, administered together and separately, on copper, zinc, calcium, and magnesium concentrations in the tissues of rats with a deficiency of both these micronutrients. The experiment was performed on 8-week-old female Wistar rats. In the first stage of the experiment, the animals were randomly assigned to a control group of rats fed the standard diet (AIN-93 M), and to a study group of rats fed a diet deficient in iron and folate. The study group was then randomly divided to four groups: group D was fed a deficit diet, group FE was fed a deficit diet with iron gluconate, the FOL group was fed a deficit diet with folate acid, and the FEFOL group was fed a deficit diet with iron gluconate and folate acid. After 2, 10, and 21 days of the intervention, ten animals from each group were killed. Mineral concentrations were assayed in the liver, spleen, pancreas, heart, and kidneys using atomic absorption spectrometry. Statistical analysis was performed using Statistica 12.0 with the ANOVA test (p

Adipose tissue is a major source of circulating exosomal microRNAs (miRNAs) that are modulators of the immune response in various types of tissues and organs, including airways. Still, no evidence exists if allergic airway inflammation may affect fat tissue inflammation via alterations in the miRNA expression profile. Therefore, we investigated the miRNA expression profile in the adipose tissue upon induced allergic inflammation in the airways in the rat. Brown Norway rats were chronically sensitized to house dust mite extract for seven weeks. Body composition was performed using MiniSpec Plus. The eosinophil count and the total IgE level were determined to confirm the induction of allergic inflammation. MiRNA expression profiling was done using the next-generation sequencing with validation by qPCR. We found that allergic airway inflammation significantly increased fat in adipose tissue, glucose concentration, and the gene expression of adipose tissue-derived proinflammatory peptides (leptin, TNF&alpha, ). In miRNA-seq analysis, we showed significant differences in the expression of 36 mature miRNAs, three precursors, and two miRNA families in adipose tissue of allergic rats. Two miRNAs&mdash, miRNA-151-5p and miRNA-423-3p&mdash, showed significantly increased expression in qPCR in adipose tissue and lungs of sensitized animals. Allergic airway inflammation affects fat tissue and alters miRNA expression profile in adipose tissue in the rat.

BACKGROUND Hedonic hunger occurs in response to a desire to consume food for pleasure. The μ-opioid system regulates the hedonic impact of food and the opioid receptor mu 1 gene (OPRM1) polymorphism has been associated with fat intake. OBJECTIVES The aim of this study was to determine whether the intake of high-fat food is associated with hedonic hunger and the OPRM1 polymorphism and whether these variables are related to BMI. METHODS Participants were 20- to 40-y-old women and men enrolled in Poznan, Poland in 2016-2018. The frequency of consumption of high-fat food was measured using a validated application for mobile devices. Hedonic hunger was assessed with the use of the Power of Food Scale (PFS). PFS1, PFS2, and PFS3 scores were generated for food available, food present, and food tasted, respectively. Genotyping of rs1799971 in the OPRM1 gene was performed using TaqMan probes. The associations were analyzed using linear regression or logistic regression, as appropriate. RESULTS Hedonic hunger scores were not associated with total high-fat food intake. Total PFS was associated with snack intake (β: 0.16, P = 0.0066). PFS1 was positively associated with healthy high-fat food intake (β: 0.27, P = 0.0001) and PFS2 with sweet high-fat food and fast-food intake (β: 0.27, P = 0.0030). OPRM1 genotype and hedonic hunger interacted on fast-food intake (β: -0.17; P

The aim of our study was to evaluate the associations between sensitivity to fat taste, eating habits and BMI value in a sample of menopausal Polish women. In a population of 95 women, fat taste thresholds with oleic acid were determined, allowing us to classify each woman as a hypersensitive or hyposensitive taster. Eating habits were assessed using a validated KomPAN questionnaire for food frequency. Dietary intake was evaluated based on a food diary. Selected biochemical parameters were measured using a Konelab20i biochemical analyzer. Anthropometric parameters and blood pressure were also measured. Twenty-two menopausal women were classified as hyposensitive to fat taste and 73 as hypersensitive. The hyposensitive tasters were significantly older (p = 0.006), with the majority of them (92%) being postmenopausal (p < 0.001); this group had significantly higher BMI values (p < 0.001) and other adiposity indicators compared to their hypersensitive counterparts. The hyposensitive tasters had higher blood pressure (systolic blood pressure; SBP p = 0.030; diastolic blood pressure; DBP p = 0.003), glucose (p = 0.011) and triacylglycerols levels than the hypersensitive tasters (p = 0.031). Almost half of them had diagnosed metabolic syndrome. Daily eating occasions were associated with low oral fatty acid sensitivity, irrespective of age (p = 0.041) and BMI value (p = 0.028). There were also significant associations between frequency of consumption of meats and eggs, as well as snacks and fast foods and low oral fatty acid sensitivity before adjustment for potential confounders (both associations p < 0.05), which remained after adjustment for age (both associations p < 0.05), but not after adjustment for BMI. A multivariate logistic regression analysis showed that higher BMI value (p = 0.003), along with postmenopausal status (p = 0.003), were associated with low fat taste sensitivity irrespective of age and consumed percentage energy from fat. Postmenopausal status and BMI are associated with low fat taste sensitivity. Fat hyposensitivity may also play a role in eating habits, leading to increased eating occasions and favoring certain types of food. These eating habits may determine increased body weight and the occurrence of metabolic syndrome in mid-life women, especially those who have undergone menopause and have been exposed to the physiological changes which are conducive to these relationships.

Spexin (SPX) is a 14 aa peptide discovered in 2007 using bioinformatics methods. SPX inhibits food intake and regulates lipid, and carbohydrate metabolism. Here, we evaluate the ability of SPX at improving metabolic control and liver function in obese and type 2 diabetic animals. The effects of 30 days SPX treatment of mice with experimentally induced obesity (DIO) or type 2 diabetes (T2DM) on serum glucose and lipid levels, insulin sensitivity and hormonal profile (insulin, glucagon, adiponectin, leptin, TNF alpha, IL-6 and IL-1β) are characterized. In addition, alterations of hepatic lipid and glycogen contents are evaluated. We report that SPX decreases body weight in healthy and DIO mice, and reduces lipid content in all three animal groups. SPX improves insulin sensitivity in DIO and T2DM animals. In addition, SPX modulates hormonal and metabolic profile by regulating the concentration of adiponectin (concentration increase) and leptin (concentration decrease) in the serum blood of DIO and T2DM mice. Lastly, SPX decreases lipid content as well as IL-6 and TNF-α protein levels in liver of DIO and T2DM mice, and reduces IL-6 and TNF-alpha concentrations in the serum derived from T2DM mice. Based on our results, we conclude that SPX could be involved in the development of obesity and type 2 diabetes mellitus and it can be further evaluated as a potential target for therapy of DIO and T2DM.