OBJECTIVE

The actions of peripherally administered <em>nesfatin</em>-<em>1</em> on glucose homeostasis remain controversial. The aim of this study was to characterize the mechanisms by which peripheral <em>nesfatin</em>-<em>1</em> regulates glucose metabolism.

METHODS

The effects of <em>nesfatin</em>-<em>1</em> on glucose metabolism were examined in mice by continuous infusion of the peptide via osmotic pumps. Changes in AKT phosphorylation and Glut4 were investigated by Western blotting and immnuofluorescent staining. Primary myocytes, adipocytes and hepatocytes were isolated from male mice.

RESULTS

Continuous peripheral infusion of <em>nesfatin</em>-<em>1</em> altered glucose tolerance and insulin sensitivity in mice fed either normal or high fat diet, while central administration of <em>nesfatin</em>-<em>1</em> demonstrated no effect. Nesfatin-<em>1</em> increases insulin secretion in vivo, and in vitro in cultured min6 cells. In addition, <em>nesfatin</em>-<em>1</em> up-regulates the phosphorylation of AKT in pancreas and min6 islet cells. In mice fed normal diet, peripheral <em>nesfatin</em>-<em>1</em> significantly increased insulin-stimulated phosphorylation of AKT in skeletal muscle, adipose tissue and liver; similar effects were observed in skeletal muscle and adipose tissue in mice fed high fat diet. At basal conditions and after insulin stimulation, peripheral <em>nesfatin</em>-<em>1</em> markedly increased GLUT4 membrane translocation in skeletal muscle and adipose tissue in mice fed either diet. In vitro studies showed that <em>nesfatin</em>-<em>1</em> increased both basal and insulin-stimulated levels of AKT phosphorylation in cells derived from skeletal muscle, adipose tissue and liver.

CONCLUSIONS

Our studies demonstrate that <em>nesfatin</em>-<em>1</em> alters glucose metabolism by mechanisms which increase insulin secretion and insulin sensitivity via altering AKT phosphorylation and GLUT 4 membrane translocation in the skeletal muscle, adipose tissue and liver.

Recently, two proteins have been localized in the arcuate nucleus (ARC) and implicated in the regulation of food intake: the serine-threonine-kinase mammalian target of rapamycin (mTOR) as part of the TOR signaling complex <em>1</em> (TORC<em>1</em>), and <em>nesfatin</em>-<em>1</em> derived from the precursor protein nucleobindin2. However, the exact cell types are not well described. Therefore, we performed double-labeling studies for NPY, CART, <em>nesfatin</em>-<em>1</em> and pmTOR in the ARC. In this study, we showed that <em>nesfatin</em>-<em>1</em> is not only intracellularly co-localized with cocaine- and amphetamine-regulated transcript (CART) peptide as reported before, but also with phospho-mTOR (pmTOR) and neuropeptide Y (NPY) in ARC neurons. Quantification revealed that 59+/-5% of the pmTOR-immunoreactive (ir) neurons were immunoreactive for <em>nesfatin</em>-<em>1</em>. Moreover, double labeling for <em>nesfatin</em>-<em>1</em> and NPY exhibited that <em>1</em>9+/-5% of the NPY positive cells were also immunoreactive for <em>nesfatin</em>-<em>1</em>. Furthermore, we could also confirm results from previous studies, showing that the majority of <em>nesfatin</em>-<em>1</em> neurons are also positive for CART peptide, whereas most of the pmTOR is co-localized with NPY and only to a lesser extent with CART.

A relationship between hormones and seizures has been reported in animals and humans. Therefore, the purpose of this study was to investigate the association between serum levels of prolactin, <em>nesfatin</em>-<em>1</em> and ghrelin measured different times after a seizure or non-epileptic event and compared with controls. The study included a total of 70 subjects, and of whom <em>1</em>8 patients had secondary generalized epilepsy (SGE), <em>1</em>6 patients had primary generalized epilepsy (PGE), <em>1</em>6 patients exhibited paroxysmal event (psychogenic) and 20 healthy males were control subjects. The first sample was taken within 5min of a seizure, with further samples taken after <em>1</em>, 24, and 48h so long as the patient did not exhibit further clinically observable seizures; blood samples were taken once from control subjects. Prolactin was measured immediately using TOSOH Bioscience hormone assays. <em>Nesfatin</em>-<em>1</em> and ghrelin peptides were measured using a commercial immunoassay kit. Patients suffering from focal epilepsy with secondary generalization and primary generalized epilepsy presented with significantly higher levels of serum prolactin and <em>nesfatin</em>-<em>1</em> and lower ghrelin levels 5min, <em>1</em> and 24h after a seizure than patients presenting with paroxysmal events (psychogenic) and control subjects; the data were similar but not statistically significant after 48h. The present study suggests that increased serum prolactin and <em>nesfatin</em>-<em>1</em> concentrations, decreased ghrelin concentrations could be used as markers to identify patients that have suffered a recent epileptic seizure or other paroxysmal event (psychogenic).

Recent studies indicate that a high level of <em>nesfatin</em>-<em>1</em>/Nucleobindin-2 (NUCB-2) is associated with poor outcome and promotes cell migration in breast cancer and prostate cancer. However, the role of NUCB2 is not well known in colon cancer. In this study, NUCB-2 level in colon cancer tissue was higher than that in non-tumor tissue. Suppression of NUCB-2 in a colon cancer cell line SW620 inhibited migration and invasion. The microarray analysis showed that low expression level of transcription factor ZEB<em>1</em> in NUCB-2 knockdowned SW620 cells. In addition, expression level of epithelial-mesenchymal transition (EMT)-related molecules including N-cadherin, E-cadherin, β-catenin, Slug and Twist was affected by NUCB-2 suppression and ZEB<em>1</em>-denepdent pathway. The signaling pathway liver kinase B<em>1</em>(LKB<em>1</em>)/AMP-dependent protein kinase (AMPK)/target of rapamycin complex (TORC) <em>1</em> was involved in regulation of NUCB-2-mediated metastasis and EMT properties. Suppression of NUCB-2 inhibited tumor nodules formation in a murine colon tumor model as well. In summary, <em>nesfatin</em>-<em>1</em>/NUCB-2 enhanced migration, invasion and EMT in colon cancer cells through LKB<em>1</em>/AMPK/TORC<em>1</em>/ZEB<em>1</em> pathways in vitro and in vivo.

<em>Nesfatin</em>-<em>1</em> is a recently discovered neuropeptide that has been shown to decrease food intake after lateral, third, or fourth brain ventricle, cisterna magna administration, or PVN injection in ad libitum fed rats. With regards to the understanding of <em>nesfatin</em>-<em>1</em> brain sites of action, additional microinjection studies will be necessary to define specific nuclei, in addition to the PVN, responsive to <em>nesfatin</em>-<em>1</em> to get insight into the differential effects on food intake. In the present study, we evaluated <em>nesfatin</em>-<em>1</em> action to modulate food intake response upon injection into the specific hypothalamic nuclei (PVN, LHA and VMN) in freely fed rats during the dark phase. We extend previous observations by showing that the <em>nesfatin</em>-<em>1</em> (50 pmol) injected before the onset of the dark period significantly reduced the <em>1</em> to 5 h cumulative food intake in rats cannulated into the PVN, LHA, but not in rats cannulated into the VMN. Glucosensing neurons located in the hypothalamus are involved in glucoprivic feeding and homeostatic control of blood glucose. In order to shed light on the mechanisms by which <em>nesfatin</em>-<em>1</em> exerts its satiety-promoting actions, we examined the effect of <em>nesfatin</em>-<em>1</em> on the excitability of hypothalamic glucosensing neurons. <em>Nesfatin</em>-<em>1</em> excited most of the glucose-inhibited (GI) neurons and inhibited most of the glucose-excited (GE) neurons in the PVN. Of 34 GI neurons in the LHA tested, inhibitory effects were seen in 70.6% (24/34) of GI neurons. The main effects were excitatory after intra-VMN administration of <em>nesfatin</em>-<em>1</em> in GE neurons (27/35, 77.<em>1</em>%). Thus, our data clearly demonstrate that <em>nesfatin</em>-<em>1</em> may exert at least a part of its physiological actions on the control of food intake as a direct result of its role in modulating the excitability of glucosensing neurons in the PVN, LHA and VMN.

OBJECTIVE

In the present study, our aim was to determine the changes in the plasma concentrations of a recently discovered peptide hormone <em>nesfatin</em>-<em>1</em> in patients with major depressive disorder and then to make a comparison with the control group.

METHODS

Subjects in the patient group were randomly selected from Mustafa Kemal University, Medical School, Research and Training Hospital, Psychiatry Department, Outpatient Clinic and subjects in the control group were selected from healthy volunteers. Healthy control subjects were matched in terms of weight and body mass index. Hamilton Depression Rating Scale (HAM-D) was applied to both groups. ELISA method was used for measurement of plasma <em>nesfatin</em>-<em>1</em> levels.

RESULTS

The average <em>nesfatin</em>-<em>1</em> level was statistically higher in patients with major depressive disorder than in the control group (p<0.00<em>1</em>). A positive correlation was observed between plasma <em>nesfatin</em>-<em>1</em> levels and HAM-D scores both in the patient group (r=0.59, p<0.00<em>1</em>) and in the control group (r=0.58, p<0.00<em>1</em>).

CONCLUSIONS

Our findings suggest a possible relationship between major depressive disorder and high plasma <em>nesfatin</em>-<em>1</em> level.

<em>Nesfatin</em>-<em>1</em>, a novel hypothalamic peptide, inhibits nocturnal feeding behavior and gastrointestinal motility in rodents. The effects of <em>nesfatin</em>-<em>1</em> on gastrointestinal secretory function, including gastric acid production, have not been evaluated. <em>Nesfatin</em>-<em>1</em> was injected into the fourth intracerebral ventricle (4V) of chronically cannulated rats to identify a <em>nesfatin</em> dose sufficient to inhibit food intake. <em>Nesfatin</em>-<em>1</em> (2 μg) inhibited dark-phase food intake, in a dose-dependent fashion, for >3 h. Gastric acid production was evaluated in urethane-anesthetized rats. <em>Nesfatin</em>-<em>1</em> (2 μg) was introduced via the 4V following endocrine stimulation of gastric acid secretion by pentagastrin (2 μg·kg(-<em>1</em>)·h(-<em>1</em>) iv), vagal stimulation with 2-deoxy-D-glucose (200 mg/kg sc), or no stimulus. Gastric secretions were collected via gastric cannula and neutralized by titration to determine acid content. <em>Nesfatin</em>-<em>1</em> did not affect basal and pentagastrin-stimulated gastric acid secretion, whereas 2-deoxy-D-glucose-stimulated gastric acid production was inhibited by <em>nesfatin</em>-<em>1</em> in a dose-dependent manner. c-Fos immunofluorescence in brain sections was used to evaluate in vivo neuronal activation by <em>nesfatin</em>-<em>1</em> administered via the 4V. <em>Nesfatin</em>-<em>1</em> caused activation of efferent vagal neurons, as evidenced by a <em>1</em>6-fold increase in the mean number of c-Fos-positive neurons in the dorsal motor nucleus of the vagus (DMNV) in <em>nesfatin</em>-<em>1</em>-treated animals vs. controls (P < 0.0<em>1</em>). Finally, <em>nesfatin</em>-induced Ca(2+) signaling was evaluated in primary cultured DMNV neurons from neonatal rats. <em>Nesfatin</em>-<em>1</em> caused dose-dependent Ca(2+) increments in 95% of cultured DMNV neurons. These studies demonstrate that central administration of <em>nesfatin</em>-<em>1</em>, at doses sufficient to inhibit food intake, results in inhibition of vagally stimulated secretion of gastric acid. <em>Nesfatin</em>-<em>1</em> activates DMNV efferent vagal neurons in vivo and triggers Ca(2+) signaling in cultured DMNV neurons.

<em>Nesfatin</em>-<em>1</em> has been identified as one of the most potent centrally acting anorexigenic peptides, and it has also been shown to play important roles in the control of cardiovascular function. In situ hybridization and immunohistochemical studies have revealed the expression of <em>nesfatin</em>-<em>1</em> throughout the brain and, in particular, in the medullary autonomic gateway known as the nucleus of the solitary tract (NTS). The present study was thus undertaken to explore the cellular correlates and functional roles of <em>nesfatin</em>-<em>1</em> actions in the medial NTS (mNTS). Using current-clamp electrophysiology recordings from mNTS neurons in slice preparation, we show that bath-applied <em>nesfatin</em>-<em>1</em> directly influences the excitability of the majority of mNTS neurons by eliciting either depolarizing (42%, mean: 7.8 ± 0.8 mV) or hyperpolarizing (2<em>1</em>%, mean: -8. 2 ± <em>1</em>.0 mV) responses. These responses were observed in all electrophysiologically defined cell types in the NTS and were site specific and concentration dependent. Furthermore, post hoc single cell reverse transcriptase polymerase reaction revealed a depolarizing action of <em>nesfatin</em>-<em>1</em> on NPY and nucleobindin-2-expressing mNTS neurons. We have also correlated these actions of <em>nesfatin</em>-<em>1</em> on neuronal membrane potential with physiological outcomes, using in vivo microinjection techniques to demonstrate that <em>nesfatin</em>-<em>1</em> microinjected into the mNTS induces significant increases in both blood pressure (mean AUC = 3354.<em>1</em> ± 750.7 mmHg·s, n = 6) and heart rate (mean AUC = <em>1</em>64.8 ± 78.5 beats, n = 6) in rats. Our results provide critical insight into the circuitry and physiology involved in the profound effects of <em>nesfatin</em>-<em>1</em> and highlight the NTS as a key structure mediating these autonomic actions.

OBJECTIVE

The mechanisms by which bariatric surgeries, including gastric bypass (GB) and sleeve gastrectomy (SG), achieve remission of type 2 diabetes mellitus (T2DM) and sustained weight reduction are unknown. We hypothesized that the novel anorexic hormone <em>nesfatin</em>-<em>1</em> and another new hormone obestatin might contribute to the marked improvement in glycemic homeostasis and weight loss in diabetics after GB and SG.

METHODS

A hospital-based, prospective study was conducted. Overnight fasting plasma concentrations of <em>nesfatin</em>-<em>1</em> and obestatin were analyzed in T2DM patients before surgery, and at 3 and <em>1</em>2 months after laparoscopic GB (n =<em>1</em>2) and SG (n = 6).

RESULTS

At <em>1</em>2 months, reductions of body mass index (BMI), fasting blood glucose, and glycated hemoglobin were similar between GB and SG groups (P all>> 0.05). Plasma <em>nesfatin</em>-<em>1</em> levels in patients undergoing GB or SG significantly decreased after surgeries (P both < 0.05). In contrast, plasma obestatin concentrations significantly increased in patients after SG (P < 0.05) but without any alteration after GB. The alterations of plasma <em>nesfatin</em>-<em>1</em> were significantly and negatively associated with the reduction of fasting blood glucose (P <0.05) at <em>1</em>2 months after GB and SG. In the SG group, the reduction of <em>nesfatin</em>-<em>1</em> significantly and positively correlated with the decrease of BMI (P < 0.05).

CONCLUSIONS

GB and SG produce differential influences with regards to circulating <em>nesfatin</em>-<em>1</em> and obestatin levels in non-morbidly obese, T2DM patients. Circulating <em>nesfatin</em>-<em>1</em> may modulate glucose homeostasis in two surgical procedures, and participate in regulating body weight in SG.

BACKGROUND

<em>Nesfatin</em>-<em>1</em>, a recently discovered anorexigenic neuropeptide, is expressed in several tissues including pancreatic islet cells and central nervous system. This peptide seems to play an important role in hypothalamic pathways regulating food intake and energy homeostasis.

OBJECTIVE

We aimed to investigate the relation of serum nesfatin-<em>1</em> level with metabolic and anthropometric parameters in obese children.

METHODS

The study included obese children with a body mass index (BMI) above the 95th percentile and healthy children with a BMI below the 85th percentile. The healthy and obese subjects had similar age and gender distribution. Fasting serum glucose, insulin, lipid profile, and nesfatin-<em>1</em> levels were measured to evaluate metabolic parameters.

RESULTS

Obese group (n = 37) had significantly higher BMI, BMI-SDS (standard deviation score of BMI), triglyceride, insulin, and insulin resistance index by the homeostasis model assessment, systolic and diastolic blood pressure values compared with the control group (n = 3<em>1</em>) (p < 0.05). Serum nesfatin-<em>1</em> level of the obese subjects was significantly lower than that of the control subjects (p = 0.00<em>1</em>). No statistically significant difference was found when nesfatin-<em>1</em> levels were compared among obese patients regarding the presence of insulin resistance (p = 0.202). In the obese group, nesfatin-<em>1</em> level was negatively correlated with BMI-SDS, but not with insulin resistance index (p = 0.02 and p = 0.36<em>1</em>, respectively).

CONCLUSIONS

This is the first study to evaluate nesfatin-<em>1</em> levels in relation with anthropometric and metabolic parameters in obese patients who had significantly lower nesfatin-<em>1</em> levels. Our results underline that nesfatin-<em>1</em> may play an important role in regulation of food intake in obese individuals.

The orexigenic peptide ghrelin and the anorexigenic peptide <em>nesfatin</em>-<em>1</em> are expressed by the same endocrine cell of the rat stomach, the X/A-like cell. However, data in humans are lacking, especially under conditions of obesity. We collected gastric tissue of obese patients undergoing sleeve gastrectomy and investigated the expression of <em>nesfatin</em>-<em>1</em> and ghrelin in the gastric oxyntic mucosa by immunofluorescence. <em>Nesfatin</em>-<em>1</em> immunoreactivity was detected in the human oxyntic mucosa in cells with an endocrine phenotype. A major portion of <em>nesfatin</em>-<em>1</em> immunoreactive cells (78 %) co-localized with ghrelin indicating the occurrence in human X/A-like cells. In patients with very high body mass index (BMI 55-65 kg/m(2)), the number of <em>nesfatin</em>-<em>1</em> immunoreactive cells/low-power field was significantly higher than in obese patients with lower BMI (40-50 kg/m(2), <em>1</em><em>1</em>8 ± <em>1</em>0 vs. 82 ± <em>1</em><em>1</em>, p < 0.05). On the other hand, the number of ghrelin immunoreactive cells was significantly reduced in obese patients with higher compared to lower BMI (96 ± <em>1</em>2 vs. 204 ± 2<em>1</em>, p < 0.0<em>1</em>). Also the ghrelin-acylating enzyme ghrelin-O-acyltransferase decreased with increasing BMI. In conclusion, <em>nesfatin</em>-<em>1</em> immunoreactivity is also co-localized with ghrelin in human gastric X/A-like cells giving rise to a dual role of this cell type with differential effects on stimulation and inhibition of appetite dependent on the peptide released. The expression of these two peptides is differentially regulated under obese conditions with an increase of <em>nesfatin</em>-<em>1</em> and a decrease of ghrelin immunoreactivity with rising BMI pointing towards an adaptive change of expression that may counteract further body weight increase.

OBJECTIVE

Millions suffer from sleep disorders that often accompany severe illnesses such as major depression; a leading psychiatric disorder characterized by appetite and rapid eye movement sleep (REMS) abnormalities. Melanin-concentrating hormone (MCH) and <em>nesfatin</em>-<em>1</em>/NUCB2 (<em>nesfatin</em>) are strongly co - expressed in the hypothalamus and are involved both in food intake regulation and depression. Since MCH was recognized earlier as a hypnogenic factor, we analyzed the potential role of <em>nesfatin</em> on vigilance.

METHODS

We subjected rats to a 72 h-long REMS deprivation using the classic flower pot method, followed by a 3 h-long 'rebound sleep'. Nesfatin mRNA and protein expressions as well as neuronal activity (Fos) were measured by quantitative in situ hybridization technique, ELISA and immunohistochemistry, respectively, in 'deprived' and 'rebound' groups, relative to controls sacrificed at the same time. We also analyzed electroencephalogram of rats treated by intracerebroventricularly administered <em>nesfatin</em>-<em>1</em>, or saline.

RESULTS

REMS deprivation downregulated the expression of <em>nesfatin</em> (mRNA and protein), however, enhanced REMS during 'rebound' reversed this to control levels. Additionally, increased transcriptional activity (Fos) was demonstrated in <em>nesfatin</em> neurons during 'rebound'. Centrally administered <em>nesfatin</em>-<em>1</em> at light on reduced REMS and intermediate stage of sleep, while increased passive wake for several hours and also caused a short-term increase in light slow wave sleep.

CONCLUSIONS

The data designate nesfatin as a potential new factor in sleep regulation, which fact can also be relevant in the better understanding of the role of nesfatin in the pathomechanism of depression.

<em>Nesfatin</em>-<em>1</em> is an anorexigenic peptide involved in energy homeostasis. Recently, <em>nesfatin</em>-<em>1</em> was reported to decrease blood glucose level and improve insulin sensitivity in high-fat diet-fed rats. However, little information is known about the influence of <em>nesfatin</em>-<em>1</em> on lipid metabolism either in physiological or diabetic condition. This study undertook whether <em>nesfatin</em>-<em>1</em> was involved in the pathophysiology in Streptozotocin-induced type 2 diabetic mice (T2DM), which was induced by a combination of high-calorie diet and two low-doses Streptozotocin. We observed that plasma <em>nesfatin</em>-<em>1</em> was significantly increased while expression of <em>nesfatin</em>-<em>1</em> neurons were decreased in hypothalamus in diabetes group compared to only high-calorie diet control group; intravenous injection of <em>nesfatin</em>-<em>1</em> decreased 0-<em>1</em>h, 0-2h, 0-3h cumulative food intake in T2DM, but 0-24h total food intake had no difference between groups. Body weight and plasma FFA were normalized after <em>nesfatin</em>-<em>1</em>(<em>1</em>0 µg/Kg) administration for 6 days. These results suggested that <em>nesfatin</em>-<em>1</em> improved lipid disorder in T2DM. It was found that blood glucose and insulin resistance coefficient decreased with treatment of <em>nesfatin</em>-<em>1</em> (both in <em>1</em> µg/Kg and <em>1</em>0 µg/Kg doses) in diabetes mice. For further understanding the role of <em>nesfatin</em>-<em>1</em> on lipid metabolism, we detected p-AMPK and p-ACC of skeletal muscle in T2DM using western blotting. The expression of p-AMPK and p-ACC increased when <em>nesfatin</em>-<em>1</em> was given with doses <em>1</em> µg/Kg but not in doses <em>1</em>0 µg/Kg. Taken together, <em>nesfatin</em>-<em>1</em> participated in the development of T2DM and stimulated free fatty acid utilization via AMPK-ACC pathway in skeletal muscle in T2DM.

<em>Nesfatin</em>-<em>1</em> is an anorexigen in goldfish. In the present study, we provide novel data indicating the presence and regulatory effects of <em>nesfatin</em>-<em>1</em> on the hypothalamo-pituitary-ovarian (HPO) axis of goldfish. Nucleobindin-2 (NUCB2)/<em>nesfatin</em>-<em>1</em>-like immunoreactive (ir) cells are present in the hypothalamus and in the pituitary, suggesting a hypophysiotropic role for <em>nesfatin</em>-<em>1</em>. NUCB2/<em>nesfatin</em>-<em>1</em>-like ir cells colocalize gonadotropin-releasing hormone (GnRH) in the nucleus lateralis tuberis posterioris and the nucleus anterior tuberis of the goldfish hypothalamus. The presence of <em>nesfatin</em>-<em>1</em> with GnRH in these two nuclei implicated in pituitary hormone release suggests a role for <em>nesfatin</em>-<em>1</em> on gonadotropin secretion. A single i.p. injection of synthetic goldfish <em>nesfatin</em>-<em>1</em> (50 ng/g body wt) resulted in an acute decrease (∼75%) in the expression of hypothalamic chicken GnRH-II and salmon GnRH mRNAs at <em>1</em>5 min postinjection in goldfish. Meanwhile, pituitary luteinizing hormone (LH) beta and follicle-stimulating hormone beta mRNAs were also inhibited (∼80%), but only at 60 min postinjection. <em>Nesfatin</em>-<em>1</em> administration also resulted in a significant reduction (∼60%) in serum LH levels at 60 min postadministration. <em>Nesfatin</em>-<em>1</em>-like immunoreactivity was also found in the follicle cells, but not the oocytes, in zebrafish and goldfish ovaries. Incubation of zebrafish follicles with <em>nesfatin</em>-<em>1</em> resulted in a significant reduction in basal germinal vesicle breakdown (∼50%) during the oocyte maturation. In addition, <em>nesfatin</em>-<em>1</em> also attenuated the stimulatory effects of maturation-inducing hormone on germinal vesicle breakdown. Together, the current results indicate that <em>nesfatin</em>-<em>1</em> is a metabolic hormone with an inhibitory tone on fish reproduction. <em>Nesfatin</em>-<em>1</em> appears to elicit this suppressive effect through actions on all three tissues in the fish HPO axis.

To assess maternal serum and cord blood apelin-36 and <em>nesfatin</em>-<em>1</em> concentrations in pregnant women with and without gestational diabetes mellitus (GDM). Thirty pregnant women with GDM and 30 gestational age matched healthy pregnant subjects participated to the study. Maternal serum and cord blood <em>nesfatin</em>-<em>1</em> and apelin-36 levels were measured with ELISA, at the time of birth. The relationships between maternal serum and cord blood <em>nesfatin</em>-<em>1</em> and apelin-36 levels, anthropometric and metabolic parameters were also assessed. Maternal serum apelin-36 levels were found higher (<em>1</em>3.5 ± 8.3 vs. 9.6 ± 5.9 ng/ml, P = 0.00<em>1</em>) and <em>nesfatin</em>-<em>1</em> levels were found lower (5.5 ± 8.<em>1</em> vs. 8.<em>1</em> ± 23.9 ng/ml, P = 0.00<em>1</em>) in patients with GDM compared with control pregnant women. However, the cord blood apelin-36 levels (8.8 ± 4.3 and 8.2 ± <em>1</em>.9 ng/ml, P = 0.6<em>1</em>8) and <em>nesfatin</em>-<em>1</em> levels (5.4 ± 4.0 and 6.2 ± <em>1</em>0.3 ng/ml, P = 0.688) were similar in the GDM and control groups, respectively. Maternal serum apelin-36 and <em>nesfatin</em>-<em>1</em> levels correlated positively with their respective cord blood levels. Maternal serum and cord blood apelin-36 levels correlated negatively with the gestational age and birth weight. Similarly maternal serum and cord blood <em>nesfatin</em>-<em>1</em> levels correlated negatively with the gestational age, but there was no correlation with the birth weight. We did not find a correlation between maternal serum apelin-36 and <em>nesfatin</em>-<em>1</em> levels, maternal age, BMI, fasting glucose, fasting insulin, and HOMA-IR. Also cord blood apelin-36 and <em>nesfatin</em>-<em>1</em> levels did not correlate with the maternal age, BMI, HOMA-IR, cord blood glucose, and cord blood insulin levels. Our results indicate that apelin-36 concentrations increase and <em>nesfatin</em>-<em>1</em> concentrations decrease in maternal serum of women with GDM.

<em>Nesfatin</em>-<em>1</em> is a novel metabolic hormone that has glucose-responsive insulinotropic actions. Islet β-cells and gastrointestinal tissues have been reported as abundant sources of <em>nesfatin</em>-<em>1</em> and its precursor hormone nucleobindin-2 (NUCB2). While <em>nesfatin</em>-<em>1</em> is emerging as a multifunctional hormone, there are no reports on the developmental expression of NUCB2/<em>nesfatin</em>-<em>1</em>. The main objective of this study was to examine the ontogenic expression of NUCB2 mRNA, and NUCB2/<em>nesfatin</em>-<em>1</em> immunoreactivity in the pancreas, stomach and duodenum, and the circulating levels NUCB2/<em>nesfatin</em>-<em>1</em> in Sprague Dawley rats. In addition, we also determined the co-localization of NUCB2/<em>nesfatin</em>-<em>1</em> and insulin immunoreactivity during development. NUCB2/<em>nesfatin</em>-<em>1</em> immunoreactivity was found in the rat stomach from postnatal days <em>1</em>3-27. Furthermore, NUCB2/<em>nesfatin</em>-<em>1</em> immunoreactivity was also detected in the enteroendocrine cells of the duodenum at postnatal days <em>1</em>3 and 27. Duodenal NUCB2 mRNA expression at postnatal day 27 was highest. Serum NUCB2/<em>nesfatin</em>-<em>1</em> levels on embryonic day 2<em>1</em> and postnatal day <em>1</em> were lower than serum NUCB2/<em>nesfatin</em>-<em>1</em> levels of adults and neonates at postnatal days <em>1</em>3, 20 and 27, gradually increasing with growth, suggesting an increase in its production and secretion from tissues including the gastrointestinal tract and pancreas. Our findings indicate that NUCB2/<em>nesfatin</em>-<em>1</em> colocalizes with insulin in the islet β-cells at all developmental stages, but the percentage of colocalization varies in an age-dependent manner. These findings suggest that NUCB2/<em>nesfatin</em>-<em>1</em> has potential age- and tissue-specific role in the developmental physiology of rats during growth.

<em>Nesfatin</em>-<em>1</em> is an 82 amino acid N-terminal fragment of nucleobindin2 that was consistently shown to reduce dark phase food intake upon brain injection in rodents. We recently reported that <em>nesfatin</em>-<em>1</em>(<em>1</em>-82) injected intracerebroventricularly (icv) reduces dark phase feeding in mice. Moreover, intraperitoneal injection of mid-fragment <em>nesfatin</em>-<em>1</em> (<em>nesfatin</em>-<em>1</em>(30-59)) mimics the food intake-reducing effects of <em>nesfatin</em>-<em>1</em>(<em>1</em>-82), whereas N-terminal (<em>nesfatin</em>-<em>1</em>(<em>1</em>-29)) and C-terminal fragments (<em>nesfatin</em>-<em>1</em>(60-82)) did not. We therefore characterized the structure-activity relationship of <em>nesfatin</em>-<em>1</em> injected icv to influence the dark phase meal pattern in mice. Mouse <em>nesfatin</em>-<em>1</em>(<em>1</em>-29), <em>nesfatin</em>-<em>1</em>(30-59), <em>nesfatin</em>-<em>1</em>(60-82) or vehicle was injected icv in freely fed C57Bl/6 mice immediately before the dark phase and food intake was monitored using an automated episodic feeding monitoring system. <em>Nesfatin</em>-<em>1</em>(30-59) (0.<em>1</em>, 0.3, 0.9 nmol/mouse) induced a dose-related reduction of 4-h food intake by 28%, 49% and 49% respectively resulting in a 23% decreased cumulative 24-h food intake compared to vehicle at the 0.3 nmol/mouse dose (p<0.05). The peak reduction occurred during the 3rd (-96%) and 4th hour (-9<em>1</em>%) post injection and was associated with a reduced meal frequency (0-4h: -47%) and prolonged inter-meal intervals (3.<em>1</em>-times) compared to vehicle (p<0.05), whereas meal size was not altered. In contrast, neither <em>nesfatin</em>-<em>1</em>(<em>1</em>-29) nor <em>nesfatin</em>-<em>1</em>(60-82) reduced dark phase food intake at equimolar doses although <em>nesfatin</em>-<em>1</em>(60-82) prolonged inter-meal intervals (<em>1</em>.7-times, p<0.05). <em>Nesfatin</em>-<em>1</em>(30-59) is the active core of <em>nesfatin</em>-<em>1</em>(<em>1</em>-82) to induce satiety indicated by a reduced meal number during the first 4h post injection. The delayed onset may be indicative of time required to modulate other hypothalamic and medullary networks regulating nocturnal feeding as established for <em>nesfatin</em>-<em>1</em>.

<em>Nesfatin</em>-<em>1</em>, an anorexigenic protein, is ubiquitously expressed in the body. However, the exact mechanism underlying the in vivo regulation of production of <em>nesfatin</em>/nucleobindin-2 (NUCB2), a precursor protein of <em>nesfatin</em>-<em>1</em>, is unknown. We investigated the influence of modulation of autonomic nerve activity by a ventromedial hypothalamus (VMH) lesion and the subsequent effect on <em>nesfatin</em>/NUCB2 production in rat tissues innervated by the peripheral nervous system. <em>Nesfatin</em>/NUCB2 is strongly expressed in the pancreas and liver, moderately expressed in subcutaneous and visceral fat tissues and interscapular brown adipose tissue (iBAT), but is weakly expressed in the skeletal muscles. Our study results showed that the VMH lesion in VMH-lesioned rats did not affect <em>nesfatin</em>/NUCB2 expression in the pancreas, liver, skeletal muscle, and iBAT; however, the protein expression was significantly high in both subcutaneous and visceral fat tissues. In addition, continuous peripheral administration of carbachol for 5 days did not affect <em>nesfatin</em>/NUCB2 expression, but chemical sympathectomy using 6-hydroxydopamine mimicked the effect of VMH lesion by showing significantly high <em>nesfatin</em>/NUCB2 expression in the subcutaneous fat tissues. These results show that VMH lesion can modulate the autonomic nervous system activity and balance and increase <em>nesfatin</em>/NUCB2 expression in white adipose tissues of rats. Further, this action may be mediated via inhibition of the sympathetic nerve activity.

<em>Nesfatin</em>-<em>1</em> was discovered in 2006 and introduced as a potential novel anorexigenic modulator of food intake and body weight. The past years have witnessed increasing evidence establishing <em>nesfatin</em>-<em>1</em> as a potent physiological inhibitor of food intake and body weight and unravelled <em>nesfatin</em>-<em>1</em>'s interaction with other brain transmitters to exert its food consumption inhibitory effect. As observed for other anorexigenic brain neuropeptides, <em>nesfatin</em>-<em>1</em> is also likely to exert additional, if not pleiotropic, actions in the brain and periphery. Recent studies established the prominent expression of the <em>nesfatin</em>-<em>1</em> precursor, nucleobindin2 (NUCB2), in the stomach and pancreas, where <em>nesfatin</em>-<em>1</em> influences endocrine secretion. This review will highlight the current experimental state-of-knowledge on the effects of NUCB2/<em>nesfatin</em>-<em>1</em> on food intake, body weight and glucose homeostasis. Potential implications in human obesity will be discussed in relation to the evidence of changes in circulating levels of NUCB2/<em>nesfatin</em>-<em>1</em> in disease states, the occurrence of genetic NUCB2 polymorphisms and--in contrast to several other hormones--the independence of leptin signalling known to be blunted under conditions of chronically increased body weight.

The recently discovered <em>Nesfatin</em>-<em>1</em> plays a role in appetite regulation as a satiety factor through hypothalamic leptin-independent mechanisms. <em>Nesfatin</em>-<em>1</em> is co-expressed with Melanin-Concentrating Hormone (MCH) in neurons from the tuberal hypothalamic area (THA) which are recruited during sleep states, especially paradoxical sleep (PS). To help decipher the contribution of this contingent of THA neurons to sleep regulatory mechanisms, we thus investigated in rats whether the co-factor <em>Nesfatin</em>-<em>1</em> is also endowed with sleep-modulating properties. Here, we found that the disruption of the brain <em>Nesfatin</em>-<em>1</em> signaling achieved by icv administration of <em>Nesfatin</em>-<em>1</em> antiserum or antisense against the nucleobindin2 (NUCB2) prohormone suppressed PS with little, if any alteration of slow wave sleep (SWS). Further, the infusion of <em>Nesfatin</em>-<em>1</em> antiserum after a selective PS deprivation, designed for elevating PS needs, severely prevented the ensuing expected PS recovery. Strengthening these pharmacological data, we finally demonstrated by using c-Fos as an index of neuronal activation that the recruitment of <em>Nesfatin</em>-<em>1</em>-immunoreactive neurons within THA is positively correlated to PS but not to SWS amounts experienced by rats prior to sacrifice. In conclusion, this work supports a functional contribution of the <em>Nesfatin</em>-<em>1</em> signaling, operated by THA neurons, to PS regulatory mechanisms. We propose that these neurons, likely releasing MCH as a synergistic factor, constitute an appropriate lever by which the hypothalamus may integrate endogenous signals to adapt the ultradian rhythm and maintenance of PS in a manner dictated by homeostatic needs. This could be done through the inhibition of downstream targets comprised primarily of the local hypothalamic wake-active orexin- and histamine-containing neurons.

Metabolic homeostasis is maintained by the coordinated action of multiple organ systems. Insulin secretion is often enhanced during obesity or insulin resistance to maintain glucose and lipid homeostasis, whereas a loss of insulin secretion is associated with type 2 diabetes. Adipocytes secrete hormones known as adipokines which act on multiple cell types to regulate metabolism. Many adipokines have been shown to influence beta cell function by enhancing or inhibiting insulin release or by influencing beta cell survival. Insulin, in turn, regulates lipolysis and promotes glucose uptake and lipid storage in adipocytes. As adipokine secretion and action is strongly influenced by obesity, this provides a potential route by which beta cell function is coordinated with adiposity, independently of alterations in blood glucose or lipid levels. In this review, I assess the evidence for the direct regulation of beta cell function by the adipokines leptin, adiponectin, extracellular nicotinamide phosphoribosyltransferase, apelin, resistin, retinol binding protein 4, fibroblast growth factor 2<em>1</em>, <em>nesfatin</em>-<em>1</em> and fatty acid binding protein 4. I summarise in vitro and in vivo data and discuss the influence of obesity and diabetes on circulating adipokine concentrations, along with the potential for influencing beta cell function in human physiology. Finally, I highlight future research questions that are likely to yield new insights into the exciting field of insulinotropic adipokines.

Rheumatic diseases encompass a diverse group of chronic disorders that commonly affect musculoskeletal structures. Osteoarthritis (OA) and rheumatoid arthritis (RA) are the two most common, leading to considerable functional limitations and irreversible disability when patients are unsuccessfully treated. Although the specific causes of many rheumatic conditions remain unknown, it is generally accepted that immune mechanisms and/or uncontrolled inflammatory responses are involved in their etiology and symptomatology. In this regard, the bidirectional communication between neuroendocrine and immune system has been demonstrated to provide a homeostatic network that is involved in several pathological conditions. Adipokines represent a wide variety of bioactive, immune and inflammatory mediators mainly released by adipocytes that act as signal molecules in the neuroendocrine-immune interactions. Adipokines can also be synthesized by synoviocytes, osteoclasts, osteoblasts, chondrocytes and inflammatory cells in the joint microenvironment, showing potent modulatory properties on different effector cells in OA and RA pathogenesis. Effects of adiponectin, leptin, resistin and visfatin on local and systemic inflammation are broadly described. However, more recently, other adipokines, such as progranulin, chemerin, lipocalin-2, vaspin, omentin-<em>1</em> and <em>nesfatin</em>, have been recognized to display immunomodulatory actions in rheumatic diseases. This review highlights the latest relevant findings on the role of the adipokine network in the pathophysiology of OA and RA.

Polycystic ovary syndrome (PCOS) is commonly characterised by obesity, insulin resistance (IR), hyperandrogenemia and hirsutism. <em>Nesfatin</em>-<em>1</em> a recently discovered hormone, acts upon energy balance, glucose metabolism, obesity and probably gonadal functions. This study was to evaluate the circulating levels of <em>nesfatin</em>-<em>1</em> in patients with PCOS (n = 30) and in age and body mass index (BMI)-matched controls (n = 30). PCOS patients had significantly lower levels of <em>nesfatin</em>-<em>1</em> (0.88 ± 0.36 ng/mL) than healthy controls (2.22 ± <em>1</em>.<em>1</em>4 ng/mL). PCOS patients also had higher gonadotropin and androgen plasma concentrations, Ferriman-Gallwey scores, blood glucose levels and a homeostasis model of assessment-IR index (HOMA-IR) index than in healthy women. Correlation tests in PCOS subjects detected a negative correlation between <em>nesfatin</em>-<em>1</em> levels and BMI, fasting blood glucose, insulin levels and a HOMA-IR index. Lower <em>nesfatin</em>-<em>1</em> concentration may plays a very important role in the development of PCOS.

BACKGROUND

Adipokines have been proved to relate with osteoarthritis (OA). As a recently discovered adipokine, <em>nesfatin</em>-<em>1</em> relationship with OA has not been reported.

OBJECTIVE

To determine the levels of <em>nesfatin</em>-<em>1</em> in serum and synovial fluid (SF) from patients with and without OA; to examine the correlation between <em>nesfatin</em>-<em>1</em> levels and high sensitivity C-reactive protein (hsCRP), Type IIA Collagen N Propeptide (PIIANP), and IL-<em>1</em>8 (interleukin-<em>1</em>8) levels in serum or synovial fluid.

METHODS

Serum and SF were collected from knee OA patients and healthy persons, respectively. Five articular tissues were obtained during TKR for immunohistochemistry (IHC). Nesfatin-<em>1</em> levels, hsCRP, PIIANP, and IL-<em>1</em>8 in serum and SF were analyzed by enzyme-linked immunosorbent assay (ELISA).

RESULTS

Nesfatin-<em>1</em> gene was expressed in OA-affected articular cartilage. OA serum contained significantly higher levels of <em>nesfatin</em>-<em>1</em>, as compared to serum from healthy controls (P < 0.05), and <em>nesfatin</em>-<em>1</em> levels in OA serum exceeded those in paired SF samples (P < 0.00<em>1</em>). Significant correlation was found between serum <em>nesfatin</em>-<em>1</em> and hsCRP levels in OA patients (r = 0.593, P = 0.00005) and also synovial <em>nesfatin</em>-<em>1</em> and IL-<em>1</em>8 levels (r = 0.560, P = 0.00<em>1</em>7).

CONCLUSIONS

Nesfatin-<em>1</em> is present in articular tissues and may contribute to the physiopathologic changes in OA. Nesfatin-<em>1</em>, accompanied with hsCRP and IL-<em>1</em>8, could be new molecular makers to speculate OA progression.