Nucleobindin-2: A Novel Regulator of Immune-Metabolic Interactions

Over the past half century, obesity has become a widespread concern to human health. Obesity is defined as having a body mass index above 30 and is accompanied by altered metabolic physiology. Obesity is one of the leading causes of mortality in the United States and in the world. The prevalence of obesity has increased over the past few decades. High body fat in adipose, excess calorie intake, low aerobic fitness are all associated with the comorbidities of obesity. Recently, an emphasis has been given to increased leukocytes in adipose tissue and the resulting low-grade systemic inflammation on the development of insulin resistance. Unfortunately, the immune-metabolic molecular mechanisms by which increased inflammation leads to diseases is not fully understood. We studied Nucb2, a highly expressed gene in immune cells and encodes for nesfatin-1, a peptide reported to be a satiety signal. We saw that Nucb2 expression is increased in leukocytes of high fat diet (HFD) fed animals. Contrary to our hypothesis, Nucb2-/- animals showed no effect on food intake. We then performed euglycemic hyperinsulinemic clamp studies in wild type and Nucb2 mice on a chow diet and a HFD to determine insulin sensitivity. Interestingly, knocking out the Nucb2 gene significantly impaired insulin sensitivity only under HFD conditions. These experiments demonstrate that Nucb2 is key player in glucose homeostasis in obesity. We discovered macrophages deficient of Nucb2 impact the proinflammatory macrophages. Macrophages lacking Nucb2 increased proinflammatory cytokine production. This suggests that Nucb2 intrinsically regulates the inflammatory cytokine production cascade. Mechanistically, Nucb2 mediates its effects by increasing proinflammatory cytokine expression via down-regulation of NFκB signaling in leukocytes. In the absence of Nucb2, NFκB activity is increased in macrophages. In contrast, inhibiting NFκB caused the opposite response. Finally, in response to endotoxemia, Nucb2 is required to control the production proinflammatory cytokines. Collectively, these data highlight a novel mechanism whereby Nucb2 serves as a key mediator of immunometabolic control of inflammation and insulin resistance. Overall, the studies I have performed identified a novel mediator of the immune-metabolic cross talks in the context of obesity-induced insulin resistance

Body composition, IGF1 status, and physical functionality in nonagenarians: implications for osteosarcopenia

OBJECTIVES: Body composition alterations occur during aging. The purpose of the present analysis was to explore the functional consequences of the overlap of sarcopenia and osteoporosis, and the potential role of insulin-like growth factor 1 (IGF1) in their development in the oldest old. SETTING AND PARTICIPANTS: Eighty-seven nonagenarians from the Louisiana Healthy Aging Study were included. MEASURES: The definition of sarcopenia was based on appendicular lean mass (ALM). Osteoporosis was diagnosed based on bone mineral density (BMD) T score. Four phenotypes were compared: (1) healthy body composition, that is, nonosteoporotic nonsarcopenic (CO, control group), (2) osteoporotic (O, low BMD T score), (3) sarcopenic (S, low ALM), and (4) osteosarcopenic (OS, low BMD T score and low ALM). Sex- and age-specific IGF1-Standard Deviation Scores (SDS) were calculated. The Continuous Scale-Physical Functional Performance (CS-PFP) test was performed. RESULTS: In OS men, IGF1-SDS values (-0.61 ±0.37 vs -0.04 ± 0.52, P = .02) were lower than those in CO males (control group), whereas IGF1-SDS were similar in the 4 body composition phenotypes in women. In men only, ALM was positively associated with IGF1-SDS values (P = .01) independent of age and C-reactive protein concentration. Regarding bone health, we found no association between IGF1-SDS values and BMD. IGF1-SDS was not associated with functional performance (CS-PFP) in men and women. CONCLUSIONS/IMPLICATIONS: IGF1 sensitivity in skeletal muscle and bone may differ by sex in the oldest old. IGF1 status did not appear to affect physical functionality. Determinants and clinical and functional characteristics of osteosarcopenia need to be further investigated in order to define conclusive diagnostic criteria

A Missing Link in Body Weight Homeostasis: The Catabolic Signal of the Overfed State

Mammals regulate fat mass so that increases or reductions in adipose tissue mass activate responses that favor return to one’s previous weight. A reduction in fat mass activates a system that increases food intake and reduces energy expenditure; conversely, overfeeding and rapid adipose tissue expansion reduces food intake and increases energy expenditure. With the identification of leptin nearly two decades ago, the central circuit that defends against reductions in body fat was revealed. However, the systems that defend against rapid expansion of fat mass remain largely unknown. Here we review the physiology of the overfed state and evidence for a distinct regulatory system, which unlike the leptin-mediated system, we propose primarily measures a functional aspect of adipose tissue and not total mass per se

Impact of Different Fecal Processing Methods on Assessments of Bacterial Diversity in the Human Intestine.

The intestinal microbiota are integral to understanding the relationships between nutrition and health. Therefore, fecal sampling and processing protocols for metagenomic surveys should be sufficiently robust, accurate, and reliable to identify the microorganisms present. We investigated the use of different fecal preparation methods on the bacterial community structures identified in human stools. Complete stools were collected from six healthy individuals and processed according to the following methods: (i) randomly sampled fresh stool, (ii) fresh stool homogenized in a blender for 2 min, (iii) randomly sampled frozen stool, and (iv) frozen stool homogenized in a blender for 2 min, or (v) homogenized in a pneumatic mixer for either 10, 20, or 30 min. High-throughput DNA sequencing of the 16S rRNA V4 regions of bacterial community DNA extracted from the stools showed that the fecal microbiota remained distinct between individuals, independent of processing method. Moreover, the different stool preparation approaches did not alter intra-individual bacterial diversity. Distinctions were found at the level of individual taxa, however. Stools that were frozen and then homogenized tended to have higher proportions of Faecalibacterium, Streptococcus, and Bifidobacterium and decreased quantities of Oscillospira, Bacteroides, and Parabacteroides compared to stools that were collected in small quantities and not mixed prior to DNA extraction. These findings indicate that certain taxa are at particular risk for under or over sampling due to protocol differences. Importantly, homogenization by any method significantly reduced the intra-individual variation in bacteria detected per stool. Our results confirm the robustness of fecal homogenization for microbial analyses and underscore the value of collecting and mixing large stool sample quantities in human nutrition intervention studies

Molecular and Physiological Adaptations to Weight Perturbation in Mice

From a medical perspective, obesity may be defined as a degree of relative adiposity sufficient to derange metabolic physiology in a manner that negatively impacts the health of the individual. While population-based cut points based on body mass index (BMI) are frequently used as a means of identifying such individuals, this is an imprecise approach since the critical levels of adiposity in this regard differ substantially among individuals. Our common genetic predisposition to increased adiposity, coupled with an environment conducive to positive energy balance results in an increasing prevalence of human obesity. Weight loss, even when initially successful, is very difficult to maintain due, in part, to a feedback system involving metabolic, behavioral, neuroendocrine and autonomic responses that are initiated to maintain somatic energy stores (fat) at a level considered `ideal' by the central nervous system (CNS). Circulating leptin is an important afferent signal to the CNS relating peripheral energy stores with modulations in key leptin sensing area sensitivity possibly implicated in the functional and molecular basis of defense of body weight. These physiological responses, which include increased metabolic efficiency at lower body weight, may be engaged in individuals at different levels of body fat depending on their genetic makeup, as well as on gestational and post-natal environmental factors that have determined the so-called "set-point". In the work presented in this dissertation the following aspects of the physiology of the defense of body weight were explored: 1) whether levels (thresholds) of defended adiposity can be raised or lowered by environmental manipulation; 2) the physiological and molecular changes that mediate increased metabolic efficiency following weight loss, 3) leptin's role in setting the threshold; 4) the effects of ambient temperature on metabolic phenotypes of weight perturbed to assess whether torpor contributes to metabolic adaptation; and 5) whether changes in gut microbiota accompany changes in diet composition and/or body weight. To assess whether the threshold for defended body weight could be increased or decreased by environmental manipulations (i.e. high fat diet and weight restriction), we identified bioenergetic, behavioral, and CNS structural responses of C57BL/6J in long term diet induced obese (DIO) male mice to weight reduction. We found that maintenance of a body weight 20% below that imposed by a high fat diet results in metabolic adaptation - energy expenditure below that expected for body mass and composition - and structural changes of synapses onto arcuate pro-opiomelanocortin (POMC) cell bodies. These changes are qualitatively and quantitatively similar to those observed in weight-reduced animals that were never obese, suggesting that the previously obese animals are now "defending" a higher body weight. Maintenance of a lower body weight for more than 3 months was not accompanied by remission of the increased metabolic efficiency. Thus, the consequence of long term elevation of body weight suggests an increase in defended body fat that does not abate with time. Mice can enter torpor - a state of decreased metabolic rate and concomitant decrease in body temperature - as a defense mechanism in times of low caloric availability and/or decreased ambient room temperatures. Declines in circulating leptin concentrations and low ambient room temperature have both been implicated in the onset of torpor. To assess the effects of ambient room temperature and leptin concentrations on metabolic adaptation, we characterized C57BL/6J and leptin deficient (Lepob) mice following weight perturbation at both 22°C and 30°C ambients. Weight-reduced C57BL/6J mice show metabolic adaptation at both ambient temperatures and do not enter torpor whereas weight-reduced Lepob animals readily enter torpor at 22°C. This suggests that sufficiently high absolute leptin concentrations may impede the onset of torpor and that torpor itself does not contribute to metabolic adaptation in mice that have an intact leptin axis. To assess whether hyperleptinemia per se was capable of increasing the threshold for defended body weight, leptin was infused by minipumps into C57BL/6J mice for 18 weeks and body weight and metabolic parameters were studied following cessation of leptin infusion. Leptin infused mice did not defend elevated body weights compared to PBS infused mice suggesting that leptin alone may not be capable of setting the threshold for body weight defense implying that other changes accompanying obesity (i.e. increased free fatty acids, endoplasmic reticulum stress and/or inflammation of leptin-sensitive neural areas) are implicated. A caveat and possible confound to this study is the possibility of antibody production against the exogenous leptin that could have drastically decreased the amount of bioavailable leptin in these mice. This experiment did not assess antibody production but subsequent studies should do so. Finally, gut microbiota have been implicated in the regulation of body weight possibly by impacting insulin resistance, inflammation, and adiposity via interactions with epithelial and endocrine cells. We assessed changes in relative abundances of cecal microbiota in mice following sustained changes in body weight and diet composition. In diet-induced obese (DIO) mice, we find that weight reduction resulted in shifts in specific bacteria abundance (Akkermansia and Mucispirillum) and that these changes were correlated with leptin concentrations. Leptin modulates mucin production in the gut possibly altering local microniches for certain bacteria providing a functional link between adiposity and gut-specific changes in bacterial populations. Overall, the major findings of these experiments are that the threshold for body weight defense can be raised but not lowered, that metabolic adaptation observed in weight-reduced mice is not a result of torpor, and that hyperleptinemia (if no anti-bodies were produced) per se isolated from other obesity-related changes does not appear capable of raising the threshold

Prevalence, awareness and control of diabetes in the Seychelles and relationship with excess body weight

BACKGROUND: The evidence for a "diabesity" epidemic is accumulating worldwide but population-based data are still scarce in the African region. We assessed the prevalence, awareness and control of diabetes (DM) in the Seychelles, a rapidly developing country in the African region. We also examined the relationship between body mass index, fasting serum insulin and DM. METHODS: Examination survey in a sample representative of the entire population aged 25-64 of the Seychelles, attended by 1255 persons (participation rate of 80.2%). An oral glucose tolerance test (OGTT) was performed in individuals with fasting blood glucose between 5.6 and 6.9 mmol/l. Diabetes mellitus (DM), impaired fasting glucose (IFG) and impaired glucose tolerance (IGT) were defined along criteria of the ADA. Prevalence estimates were standardized for age. RESULTS: The prevalence of DM was 11.5% and 54% of persons with DM were aware of having DM. Less than a quarter of all diabetic persons under treatment were well controlled for glycemia (HbA1c), blood pressure or LDL-cholesterol. The prevalence of IGT and IFG were respectively 10.4% and 24.2%. The prevalence of excess weight (BMI > or = 25 kg/m2) and obesity (BMI > or = 30 kg/m2) was respectively 60.1% and 25.0%. Half of all DM cases in the population could be attributed to excess weight. CONCLUSION: We found a high prevalence of DM and pre-diabetes in a rapidly developing country in the African region. The strong association between overweight and DM emphasizes the importance of weight control measures to reduce the incidence of DM in the population. High rates of diabetic persons not aware of having DM in the population and insufficient cardiometabolic control among persons treated for DM stress the need for intensifying health care for diabetes