Werner syndrome (WS) is an autosomal recessive disorder associated with premature aging and cancer predisposition. WS cells show increased genomic instability and are hypersensitive to DNA-damaging agents. WS is caused by mutations of the WRN gene. WRN protein is a member of RecQ DNA helicase family. In addition to a conserved 3’–5’ helicase activity, the WRN protein contains unique 3’–5’ exonuclease activity. WRN recognizes specific DNA structures as substrates that are intermediates of DNA metabolism. WRN physically and functionally interacts with many other proteins that function in telomere maintenance, DNA replication, and DNA repair. The function of WRN is regulated by post–translational modifications that include phosphorylation, acetylation, and sumoylation. SIRT1 is a NAD-dependent histone deacetylase (HDAC) that deacetylates histones and a numbers of cellular proteins. SIRT1 regulates the functions of many proteins, which are important for apoptosis, cell proliferation, cellular metabolism, and DNA repair. SIRT1 is also regulated by other proteins or molecules from different levels to activate or inhibit its deacetylase activity. In this study, we found that SIRT1 interacts with and deacetylates WRN. We further identified the major acetylation sites at six lysine residues of the WRN protein and made a WRN acetylation mutant for functional analysis. We found that WRN acetylation increases its protein stability. Deacetylation of WRN by SIRT1 reverses this effect. CREB-binding protein (CBP) dramatically increased the half-life of wild-type WRN, while this increase was abrogated with the WRN acetylation mutant. We further found that WRN stability is regulated by the ubiquitination pathway, and that WRN acetylation by CBP dramatically reduces its ubiquitination level. We also found that acetylation of WRN decreases its helicase and exonuclease activities, and that SIRT1 reverses this effect. Acetylation of WRN alters its nuclear distribution. Down-regulation of SIRT1 increases WRN acetylation level and prevents WRN protein translocating back to nucleolus after DNA damage. Importantly, we found that WRN protein is strongly acetylated and stabilized in response to mitomycin C (MMC) treatment. H1299 cells that were stably expressing WRN acetylation mutant display significantly higher sensitivity to MMC than the cells expressing wild-type WRN. Taken together, these data demonstrated that acetylation pathway plays an important role in regulating WRN function in response to DNA damage. A model has been proposed based on our discoveries.

Background Obesity is an important risk factor for resistance to insulin-mediated glucose disposal, and is a precursor of type 2 diabetes and other disorders. Objectives To identify molecular pathways in adipose tissue and inflammatory cells that may result in obesity-associated insulin resistance, we exploited the fact that not all obese individuals are prone to insulin resistance. Thus the degree of obesity as a variable was removed by studying obese subjects of similar body mass index (BMI) who are insulin-sensitive (IS) versus insulin-resistant (IR). Methods Combining gene expression profiling with computational approaches, we determined the global gene expression signatures of omental and subcutaneous adipose tissue samples obtained from 10 obese-IR and 10 obese-IS patients undergoing gastric bypass surgery. In a secondary study, we isolated monocytes from 4 obese-IR, 3 obese-IS, and 4 nonobese-IS adolescent and young adult subjects for purposes of assessing differences in expression of inflammatory genes in monocytes using RT-PCR. Results Gene sets related to chemokine activity and chemokine receptor-binding were identified as most highly enriched in the omental tissue from obese-IR compared to obese-IS subjects, independent of BMI. Strikingly, insulin resistance, but not BMI, was associated with increased macrophage infiltration in the omental adipose tissue, as was adipocyte size. In the adolescent and young adult cohort, expression of two cytokine signaling molecules (IL8, SOCS3) and two downstream products of the JNK pathway (JunB, c-Fos) showed increased expression in the obese-IR subjects compared to the obese-IS and nonobese-IS subjects, suggesting the presence of a proinflammatory phenotype in monocytes in obesity, which is exacerbated in the insulin resistant state. Conclusions Our findings demonstrate that inflammation of omental adipose tissue and activation of proinflammatory monocytes is strongly associated with insulin resistance in human obesity. Manipulation of these pathways may result in the prevention of or delay in the onset of obesity-related co-morbidities.

L’obésité est un facteur de risque important du diabète de type 2 et des maladies cardio- vasculaires. Durant le développement de l’obésité, le remodelage pathologique du tissu adipeux et l’inflammation locale contribuent à la mise en place d’une inflammation systémique, une accumulation de gras ectopique et une résistance à l’insuline. Les macrophages jouent un rôle important dans la régulation des voies signalétiques inflammatoires qui sont liées à leurs différents états d’activation. En effet, l’influence de diverses adipokines, cytokines et hormones ainsi que la disponibilité des nutriments vont induire leur polarisation en un phénotype soit pro- inflammatoire M1, soit anti-inflammatoire M2. De ce fait, comprendre les mécanismes et acteurs impliqués dans la polarisation des macrophages résidents du tissu adipeux vers un phénotype anti-inflammatoire M2 pourrait apporter une stratégie pour traiter les complications de l’obésité. La suppression de l’α/β-hydrolase domain-6 (ABHD6), une monoacylglycérol lipase, a démontré chez la souris des effets bénéfiques contre l’obésité, le diabète de type 2 et d’autres maladies inflammatoires. Cependant, le rôle précis de l’ABHD6 dans l’activation et la polarisation des macrophages en conditions inflammatoires, ainsi que les mécanismes sous-jacents, ne sont pas clairement définis. Le but de cette étude était d’investiguer : 1) le rôle immuno-métabolique de l’ABHD6 dans l’inflammation chronique du tissu adipeux induite par l’obésité chez la souris, et 2) le rôle régulateur de l’ABHD6 dans l’activation et la polarisation des macrophages dans des conditions inflammatoires aiguës induite par le lipopolysaccharide (LPS). En employant des approches pharmacologiques et génétiques, nous avons démontré que la délétion globale de l’ABHD6, chez les souris rendues obèses par une diète riche en gras, permet de maintenir un tissu adipeux sain au niveau immuno-métabolique. En effet, les souris ABHD6-KO nourries avec une diète riche en gras montrent une expression réduite de l’expression des marqueurs pro- inflammatoires (Mcp1 and Cd11c), fibrotiques (Col5a1) et hypoxiques (Hif1a) dans tous leurs tissus adipeux (viscéral, sous-cutané et brun). Le nombre de macrophages pro-inflammatoires M1 est aussi diminué dans les tissus adipeux viscéral et brun des souris KO nourries à la diète grasse. De plus, la suppression de l’ABHD6 a changé la polarisation des macrophages d’un phénotype pro-inflammatoire M1 vers un phénotype anti-inflammatoire M2 dans des lignées cellulaires de macrophages (RAW264.7 et J774A.1) et des macrophages péritonéaux primaires de souris traités avec le LPS. Collectivement, nos résultats supportent la vision que l’ABHD6 a un rôle pro-inflammatoire dans induit par la diète riche en gras ou le LPS. Ces observations nous ont permis d’obtenir un aperçu du rôle de l’ABHD6 dans les voies immuno-métaboliques des tissus adipeux et suggèrent que l’ABHD6 pourrait être une cible thérapeutique intéressante pour contrer l’inflammation et les complications de l’obésité. / Obesity is a major risk factor for type 2 diabetes (T2D) and cardiovascular diseases. Pathologic adipose tissue (AT) remodeling and local inflammation contribute to systemic inflammation, ectopic fat accumulation and insulin resistance in obesity. Macrophages play an important role in inflammatory pathways, and these cells possess different activation states, M1 pro-inflammatory and M2 anti-inflammatory, influenced by adipokines, cytokines, hormones, and fuel availability. Thus, understanding the mechanisms and players involved in AT-resident macrophage polarization towards an anti-inflammatory M2 phenotype may provide a strategy to treat obesity complications. Suppression/deletion of the monoacylglycerol lipase α/β-hydrolase domain-6 (ABHD6) was previously shown to have beneficial effects against obesity, T2D, and other inflammatory disorders. However, it is not precisely known if ABHD6 plays any direct role in macrophage activation and polarization under inflammatory conditions, and if it does, the underlying mechanisms are yet to be defined. The aim of this study is to investigate: 1) the immunometabolic role of ABHD6 in obesity-induced chronic AT inflammation in mice, and 2) the regulatory role of ABHD6 in macrophage activation/polarization under conditions of acute lipopolysaccharide (LPS)-induced inflammation. Employing pharmacological and genetic approaches, we demonstrate that global ABHD6 deletion maintains AT immunometabolic health during diet-induced obesity in mice. High-fat-diet(HFD)-fed whole-body ABHD6-KO (HFD-ABHD6- KO) mice exhibit lower expression of pro-inflammatory (Mcp1 and Cd11c), fibrosis (Col5a1) and hypoxia (Hif1a) markers in all fat depots. The pro-inflammatory M1-macrophages in the visceral and brown fat depots are also reduced in HFD-ABHD6-KO mice. Additionally, ABHD6 suppression switches LPS-induced macrophage polarization from an M1-like phenotype towards an anti- inflammatory M2 state in the macrophage cell lines (RAW264.7 and J774A.1) and in primary peritoneal macrophages. Collectively, our results support the view that ABHD6 has a pro-inflammatory role under conditions of HFD- and LPS-induced inflammation. These findings provide an insight into the role of ABHD6 in AT immunometabolic pathways in obesity, and suggest ABHD6 as a therapeutic target for inflammation and obesity-related complications.

Purpose: This qualitative descriptive study was undertaken to describe Eastern Woodlands Native adult perspectives, health care beliefs and type 2 diabetes management experiences. Specific Aims: The specific aims were to 1) explore and describe perceptions of type 2 diabetes among Eastern Woodland Native adults and how they relate to their understandings about the cause and treatment approaches to the disease, 2) describe how family, friends, and community intersect with type 2 diabetes management, 3) describe relationships with health care providers and 4) determine resources that would help diabetes-self management within their community. Framework: The PEN-3 Model by Airhihenbuwa was the initial framework used for the study. Methods: A qualitative descriptive design with maximum variation and snowball sampling was used and data was analyzed using qualitative content analysis. Results: The overarching theme of Together We Can Return To Balance comprised five sub-themes: Coming to Know Life Paths with T2DM, Negotiating My Way Forward, Making Important Connections, Acknowledging the Imbalance, and Sticking Closer to Mother Earth illustrating physical, spiritual, and environmental health factors influencing DSM capacities. Conclusion: Native perspectives should be viewed as a crucial contextual variation for type 2 diabetes care when developing DSMES and for improving DSM capacities in these populations.

Obesity accounts for $168 billion in annual medical expenses and increases the risk of cardiovascular disease, cancer, and type-2 diabetes, three diseases responsible for over 50% of deaths in the United States. It is well established that the pattern of adiposity is an important factor in the relationship with disease risk and that visceral adiposity, which favors hypertrophy (characterized by enlarged cells) is more dangerous than subcutaneous adiposity, which tends to be hyperplastic (characterized by an increase in cell number). Hypertrophy is associated with inflammation and insulin resistance, and hyperplasia (adipogenesis, i.e., the formation of new adipocytes), is associated with improved insulin sensitivity. Tumor necrosis factor-alpha (TNF-alpha) is a potent pro-inflammatory cytokine that activates a nuclear factor-kappa B (NFKB) intracellular pathway that is an important mediator of obesity-associated insulin resistance and increased risk of type-2 diabetes. Interestingly, obesity has been positively associated with both low vitamin D status and elevated levels of TNF-alpha. Our studies focused on examining the influence of the active vitamin D hormone, 1,25-dihydroxyvitamin D, and TNF-alpha on adipogenesis and inflammation in human primary adipocytes and determining whether the balance of these two factors influences the extent to which adipocytes accumulate lipid or express pro-inflammatory cytokines. We found no effect of 1,25-dihydroxyvitamin D on adipogenesis or pro-adipogenic gene expression despite a clear upregulation of a vitamin D responsive gene, 24-hydroxylase, in response to treatment with 1,25-dihydroxyvitamin D. TNF-alpha clearly inhibited adipogenesis and expression of PPAR-gamma and C/EBP-alpha and enhanced expression of the pro-inflammatory cytokines IL-6 and MCP-1, but not IL-8. There was a trend towards a dose-dependent downregulation of MCP-1 by 1,25-dihydroxyvitamin D in three individuals; however, this effect was not statistically significant. While we found no interaction between TNF-alpha and 1,25-dihydroxyvitamin D on adipogenesis, there is a potential anti-inflammatory action of 1,25-dihydroxyvitamin D in human primary adipocytes. Future studies into this potential are warranted in light of the growing obesity epidemic and the interest in finding nutritionally modifiable treatment or prevention strategies to mitigate the negative consequences of obesity.

Use of biochemical markers (Lipoprotein phospholipase A2 and hyaluronic acid) for laboratory diagnosis of metabolic and degenerative diseases of the musculoskeletal system Abstract Musculoskeketal disorders currently belong to the most common diseases. The presented work describes the use of biochemical markers (Lipoprotein phospholipase A2 and hyaluronic acid) for laboratory diagnosis of metabolic and degenerative diseases of the musculoskeletal system. Concentrations of LP-PLA2 were significantly elevated in the patients with bone resorption compared to the control group of healthy individuals. Serum levels of Lp-PLA2 also negatively correlated with decreased levels of serum osteocalcin in patients. HA concentrations in synovial fluid did not differ from published reference values in synovial fluid. Patients who underwent arthroscopy had significantly elevated synovial HA concentration than patients who underwent total knee endoprosthesis. HA positively correlates with osmotic pressure determined by examination of osmolality in synovial fluid. Lipoprotein phospholipase A2 concentrations are elevated in patients with bone density impairment. LpPLA2 concentrations correlate with the severity of bone density impairment expressed by the T score. Hyaluronic acid concentrations in patients with knee...

Background: A diet rich in vegetables, fruits, grains, fat-free or low-fat dairy, and proteins is known to have multiple beneficial health effects. However, a very limited number of studies have characterized food group intake and its association with cardiometabolic risk factors in Hispanic children. Objectives: The objectives of this study were to 1) assess food group intake in a sample of Hispanic children; and 2) examine the association between food group intake and overweight and elevated blood pressure in the same population. Methods: Data came from a pilot study of metabolic syndrome in Hispanic children. The study sample consisted of 116 2-to-10-year-old children receiving well-child care at a community health center in Johnson City, TN, from June 2015 to June 2016. Blood pressure, height and weight were measured using standard protocols. Food group intake was ascertained using the Block Kids Food Screener, a validated instrument. Child’s age, sex, and mother’s educational attainment were included as covariates. Binomial test of proportions was used to compare the study sample with the National Health and Nutrition Examination Survey (NHANES) gender-age group proportions not meeting recommended daily food group intake. Two sample t-test was used to examine differences in mean food intake by outcome variables of elevated blood pressure (>=90th percentile for age and sex) and being overweight (>85th percentile of the 2000 CDC growth charts). Multiple logistic regression was used to examine the association between food group intake and elevated blood pressure and being overweight while accounting for child's sex, age, and mother's educational attainment. Results: Hispanic children exceeded minimum fruit and legume national recommendations. Compared with the corresponding NHANES gender-age groups, a larger proportion of the sample met legume recommendations. However, similar proportions met fruit, vegetable, wholegrain, fiber and dairy recommendations. Children with elevated blood pressure ate less fruits, vegetables, and legumes than children with normal blood pressure. Legume intake (OR: 0.052, 95% CI: 0.04-0.64), dairy intake (OR: 0.61, 95% CI: 0.37-0.99), and fiber intake (OR: 0.88, 95% CI: 0.81-0.96) were protective against elevated blood pressure. In contrast, only fruit intake was protective against overweight (OR: 0.93, 95% CI: 0.87-0.99). Conclusion: Public health nutrition programs aimed at reducing the prevalence of overweight and elevated blood pressure in Hispanic children should consider supporting the intake of legumes, dairy, and fiber (for decreasing elevated blood pressure), and fruits (for reducing overweight).

Maternal obesity, physical inactivity, and negative perceptions of neighborhoods have been identified as obesogenic factors in older children; however, no study has explored this relationship in young Hispanic children. Furthermore, the relation between obesogenic maternal factors and blood pressure in Hispanic children has not been examined. Objectives: This study aimed to assess the association between Hispanic mother’s physical activity (PA) levels, body mass index (BMI), and PA-related perceptions of neighborhoods and their children’s PA, TV screen time, blood pressure (BP), and BMI. Methods: Data of 118 mother-child dyads enrolled in a cross-sectional study of metabolic syndrome in Hispanic children at a community health center in Johnson City, TN were used. Parent and child questionnaires were used to ascertain mother’s BMI, PA, perception of the safety and availability of PA amenities in their children’s neighborhoods, satisfaction with their children’s neighborhoods as a place to bring up children, and children’s PA and TV screen time. Children’s height, weight, and BP were measured. Multiple logistic regression was used to examine the association between child and maternal variables, adjusting for mother’s education and the child’s sex and age. Results: Children of obese mothers were more likely than children of mothers with normal BMI to engage in less than three days of at least 60 minutes of vigorous PA per week (OR: 6.47: 95% CI: 1.61-26.0). Children whose mothers did not engage in moderate PA were more likely to engage in less than three days of at least 60 minutes of vigorous PA per week (OR: 2.92, CI: 1.18-7.24); and have elevated BP (OR: 2.50, 95% CI: 1.02-4.53) than children whose mothers engaged in moderate PA. Conclusion: Our results suggest the need for interventions to help Hispanic mothers model physical activity and achieve and maintain healthy weight; if successful, these strategies could potentially influence their children’s blood pressure and physical activity levels.

As the prevalence of vitamin D deficiency continues to grow, mounting evidence supporting its link with chronic disease strengthens suggesting vitamin D’s candidacy in the prevention and treatment of multiple disease states and their complications. Dietary guidelines, however, do not take sun exposure into account. The present study sought to explore the impact of sun exposure on vitamin D status (serum 25(OH)D), and identify other significant determinants of serum levels which may have the greatest effects on overall health. Participants (n = 34) were pre-menopausal women aged 18 to 50 years (mean age 39 ± 6 years), who had their blood drawn at a local pathology lab and a follow-up appointment at a health assessment lab for the collection of other measurements. Mean serum 25(OH)D level was 64 ± 18 nmol/L, and mean dietary vitamin D intake was approximately 327 ± 229 IU/day. Although 82% of participants were below the RDA guidelines (600 IU/day for females ages 9-50 years) for dietary vitamin D intake, only 32% had serum 25(OH)D levels < 50 nmol/L (the recommended level of sufficiency for bone health) reflecting deficiency. While serum 25(OH)D levels were significantly correlated to dietary vitamin D intake (r = 0.42, p = 0.0139), it is reasonable to assume that participants obtained adequate vitamin D from sun exposure. Fasting serum insulin levels were significantly, positively correlated with BMI (r = 0.83, p < 0.0001), and sun exposure index (Body Surface Area x Minutes of Direct Sunlight) was significantly, positively correlated with serum 25(OH)D levels (fall weekend SEI: r = 0.47, p = 0.0059; spring weekend SEI: r = 0.43, p = 0.0135; average weekend SEI: r = 0.43, p = 0.013; and average overall SEI: r = 0.39, p = 0.0247). Reported sun exposure appeared to be least during winter weekdays and the most during summer weekends. Regression analysis was used to determine the strongest predictors of serum 25(OH)D levels, which were found to be sun exposure, dietary vitamin D intake, skin reflectance, age, BMI, and ethnicity (R2 = 0.58 , p = 0.0031), demonstrating that simple questionnaires, such as those employed in this study, can help to predict serum 25(OH)D status and thus be considered in the future treatment of vitamin D deficiency.

The prevalence of non-alcoholic fatty liver disease (NAFLD) in children has increased over the past decades, creating a need for animal models that recapitulate the features of the pediatric disease. Iberian pigs have a leptin-resistant phenotype characterized by hyperleptinemia, hyperphagia, and extreme adipogenesis. We hypothesized that neonatal Iberian pigs fed a high fat high-fructose (HFF) diet will develop a pattern of liver injury resembling pediatric NAFLD. In addition, we sought to determine if a mixture of probiotics would prevent the disease. Animals were fed 1 of 4 diets containing (g/kg body weight × d) 0 g fructose, 11 g fat and 199 kcal (CON-N; n=8), 22 g fructose, 16 g fat and 300 kcal (HFF2-N; n=8), CON + probiotic (CON-P; n=6), or HFF2 + probiotic (HFF2-P; n=6) every 6 h for 70 d. The probiotic mixture (6.2 × 104 cfu/mL) contained Pediococcus acidilactici, Pediococcus pentosaceus, Lactobacillus plantarum and Bacillus amyloliquefaciens. Body weight was recorded every 3 d. Serum markers of liver injury and dyslipidemia were measured on d 40 and 65 at 2 h post feeding. Fasting leptin, insulin, glucose and homeostatic model assessment (HOMA) values were assessed on d 70. Liver and skeletal muscle (longissimus dorsi) were collected on d 70 for histology, triacylglyceride (TAG) quantification, relative gene expression, and Western blot analysis. Metabolomic analysis was performed on liver tissue and plasma. Body weight was not significantly greater in HFF fed pigs compared to CON. Leptin, alanine and aspartate aminotransferases, alkaline phosphatase, lactate dehydrogenase and total bilirubin were increased (P ≤ 0.001), and high and low density lipoproteins decreased (P ≤ 0.05) in HFF2-N and HFF2-P. Livers in HFF2-P and HFF2-N had higher relative weight and TAG (P ≤ 0.001), micro and macrovesicular steatosis, ballooning degeneration, Mallory-denk bodies, inflammation and necrosis, increased gene expression of TNFα, TGFβ, IL1α and PPARγ (P ≤ 0.001), and decreased ChREBP (P ≤ 0.001). A probiotic affect was seen as pigs fed CON-P and HFF2-P had higher insulin and HOMA values were increased (P ≤ 0.05). Western blot analysis showed dysregulation of autophagy in liver of pigs fed CON-P and HFF2-P, and in skeletal muscle of pigs fed CON-N and HFF2-N. Metabolomic analysis demonstrated dysregulation of one-carbon metabolism, the tricarboxylic acid cycle (TCA), the urea cycle, and amino acid metabolism of pigs fed HFF2 diets compared to CON diets. In conclusion, Iberian pigs fed a HFF diet recapitulate many pediatric NAFLD-associated features, in the absence of obesity and independently of probiotic supplementation, suggesting a potentially suitable model for pediatric NAFLD research. Furthermore, probiotic supplementation did not ameliorate the onset of NAFLD when fed in conjunction with a HFF diet.