<em>Nesfatin</em>-<em>1</em> is a recently discovered anorexigenic peptide which is distributed in several brain areas implicated in the feeding and metabolic regulation. Recently, it has been reported that <em>nesfatin</em>-<em>1</em> is expressed not only in brain, but also in peripheral organs such as digestive organs, adipose tissues, heart, and reproductive organs. <em>Nesfatin</em>-<em>1</em> is markedly expressed in the pancreas, stomach and duodenum. Eventually, the <em>nesfatin</em>-<em>1</em> expression in the digestive organs may be regulated by nutritional status, which suggests a regulatory role of peripheral <em>nesfatin</em>-<em>1</em> in energy homeostasis. <em>Nesfatin</em>-<em>1</em> is also detected in the adipose tissues of humans and rodents, indicating that <em>nesfatin</em>-<em>1</em> expression in the fat may regulate food intake independently, rather than relying on leptin. In addition, <em>nesfatin</em>-<em>1</em> is expressed in the heart as a cardiac peptide. It suggests that <em>nesfatin</em>-<em>1</em> may regulate cardiac function and encourage clinical potential in the presence of nutrition-dependent physio-pathologic cardiovascular diseases. Currently, only a few studies demonstrate that <em>nesfatin</em>-<em>1</em> is expressed in the reproductive system. However, it is not clear yet what function of <em>nesfatin</em>-<em>1</em> is in the reproductive organs. Here, we summarize the expression of <em>nesfatin</em>-<em>1</em> and its roles in brain and peripheral organs and discuss the possible roles of <em>nesfatin</em>-<em>1</em> expressed in reproductive organs, including testis, epididymis, ovary, and uterus. We come to the conclusion that <em>nesfatin</em>-<em>1</em> as a local regulator in male and female reproductive organs may regulate the steroidogenesis in the testis and ovary and the physiological activity in epididymis and uterus.

<em>Nesfatin</em>-<em>1</em> (82 amino acid) is an anorexigenic and insulinotropic peptide encoded in a secreted precursor, nucleobindin-2 (NUCB2). Nucleobindin-<em>1</em> (NUCB<em>1</em>) is a protein with very high sequence similarity to NUCB2. We hypothesized that a <em>nesfatin</em>-<em>1</em> like peptide (NLP) is encoded in NUCB<em>1</em>, and this peptide is biologically active. In silico analysis found a signal peptide cleavage site at position 25 (Arginine) and 26 (Valine) preceding the NLP region in NUCB<em>1</em> sequence, and potential proprotein convertase cleavage sites at Lys-Arg (KR), forming a 77 amino acid NLP. RT-PCR studies found NUCB<em>1</em> mRNA in both pancreas and MIN6 cells. NUCB<em>1</em>-like immunoreactivity was detected in mouse insulinoma (MIN6) cells, and pancreatic islet beta cells of mice. In order to determine the biological activity of NLP, MIN6 cells were incubated with synthetic rat NLP. NLP (<em>1</em>0nM and <em>1</em>00nM) upregulated preproinsulin mRNA expression and insulin secretion at <em>1</em>h post-incubation. In identical experiments using MIN6 cells, a scrambled peptide based on the NLP sequence did not elicit any effects on preproinsulin mRNA expression or insulin secretion. From this result, it is clear that an intact NLP sequence is required for its biological activity. NLP appears as another endogenous insulinotropic peptide encoded in NUCB<em>1</em>.

BACKGROUND

Hashimoto's thyroiditis is an autoimmune disorder and the most common cause of hypothyroidism. The use of Nigella sativa, a potent herbal medicine, continues to increase worldwide as an alternative treatment of several chronic diseases including hyperlipidemia, hypertension and type 2 diabetes mellitus (T2DM). The aim of the current study was to evaluate the effects of Nigella sativa on thyroid function, serum Vascular Endothelial Growth Factor (VEGF) - <em>1</em>, <em>Nesfatin</em>-<em>1</em> and anthropometric features in patients with Hashimoto's thyroiditis.

METHODS

Forty patients with Hashimoto's thyroiditis, aged between 22 and 50 years old, participated in the trial and were randomly allocated into two groups of intervention and control receiving powdered Nigella sativa or placebo daily for 8 weeks. Changes in anthropometric variables, dietary intakes, thyroid status, serum VEGF and <em>Nesfatin</em>-<em>1</em> concentrations after 8 weeks were measured.

RESULTS

Treatment with Nigella sativa significantly reduced body weight and body mass index (BMI). Serum concentrations of thyroid stimulating hormone (TSH) and anti-thyroid peroxidase (anti-TPO) antibodies decreased while serum T3 concentrations increased in Nigella sativa-treated group after 8 weeks. There was a significant reduction in serum VEGF concentrations in intervention group. None of these changes had been observed in placebo treated group. In stepwise multiple regression model, changes in waist to hip ratio (WHR) and thyroid hormones were significant predictors of changes in serum VEGF and Nesgfatin-<em>1</em> values in Nigella sativa treated group (P < 0.05).

CONCLUSIONS

Our data showed a potent beneficial effect of powdered Nigella sativa in improving thyroid status and anthropometric variables in patients with Hashimoto's thyroiditis. Moreover, Nigella sativa significantly reduced serum VEGF concentrations in these patients. Considering observed health- promoting effect of this medicinal plant in ameliorating the disease severity, it can be regarded as a useful therapeutic approach in management of Hashimoto's thyroiditis.

BACKGROUND

Iranian registry of clinical trials (registration number IRCT20<em>1</em>502<em>1</em>7<em>1</em>9082N4 - Registered March-<em>1</em>5-20<em>1</em>5).

OBJECTIVE

An imbalance in the production of adipokines and myokines impairs the energy expenditure, increases adipocyte and develops metabolic pathologies. Physical exercise is able to regulate the secretion of myokines and adipokines. The present study considers the metabolic cross talk between skeletal muscle and adipose tissue in high-intensity interval training vs. moderate-intensity continuous training by regulation of PGC-<em>1</em>α.

METHODS

A sample of 32 male Wistar rats (8 weeks old with mean weight 250 ± 55 g) were divided into four groups randomly: control of base (CO), control of 8 weeks (CO8w), moderate-intensity continuous training (MICT), and high-intensity interval training (HIIT). The rats were fed with standard chow diet. The CO group was killed at the start of the study and the CO8w group was kept alive for the same time as the experimental groups, but did not participate in any exercise. MICT and HIIT groups for 8 weeks were placed under the moderate-intensity continuous training (<em>1</em>5-60 min, with speed of <em>1</em>5-30 m/min) and high-intensity interval training (8-4 intense period for <em>1</em> min, with speed of 28-55 m/min, with 3-7 slow-intensity period for <em>1</em> min, with a speed of <em>1</em>2-30 m/min) for 8 weeks, respectively. To measure the levels of serum irisin, <em>nesfatin</em>, and resistin the ELISA method was used and real-time PCR method was used to evaluate the relative expression of soleus PGC-<em>1</em>α gene mRNA.

RESULTS

The levels of irisin and <em>nesfatin</em> significantly increased in the HIIT compared with control groups (p = 0.00<em>1</em>). Resistin values in both training groups showed a significant decrease compared to the control groups (p = 0.005). The level of PGC-<em>1</em>α gene expression in both HIIT and MICT groups was significantly increased in comparison with the control groups (p = 0.00<em>1</em>).

CONCLUSIONS

The results showed that HIIT and MICT increase the transcription of the PGC-<em>1</em>α gene and possibly the increased expression of this gene after HIIT and MICT plays a central role in the secretion of skeletal muscle myokines and adipokines of adipose tissue.

METHODS

No Level of evidence: Animal study.

<em>Nesfatin</em>-<em>1</em> is a recently identified anorexigenic peptide mainly secreted from the brain and adipose tissue. Although <em>nesfatin</em>-<em>1</em> may have pro-inflammatory and apoptotic properties, the association between plasma <em>nesfatin</em>-<em>1</em> levels and coronary artery disease (CAD) has not been clarified yet. We investigated plasma <em>nesfatin</em>-<em>1</em> levels in 302 patients undergoing elective coronary angiography. Of the 302 study patients, CAD was present in <em>1</em>72 (57%), of whom 67 had <em>1</em>-vessel, 49 had 2-vessel, and 56 had 3-vessel disease. Compared with <em>1</em>30 patients without CAD, <em>1</em>72 with CAD had higher plasma <em>nesfatin</em>-<em>1</em> levels (median 0.2<em>1</em> vs. 0.<em>1</em>7 ng/mL, P < 0.0<em>1</em>). A stepwise increase in <em>nesfatin</em>-<em>1</em> levels was found depending on the number of > 50% stenotic coronary vessels: 0.<em>1</em>7 in CAD(-), 0.20 in <em>1</em>-vessel, 0.2<em>1</em> in 2-vessel, and 0.22 ng/mL in 3-vessel disease (P < 0.05). A high <em>nesfatin</em>-<em>1</em> level (> 0.<em>1</em>9 ng/mL) was found in 43% of patients with CAD(-), 55% of those with <em>1</em>-vessel, 55% of those with 2-vessel, and 68% of those with 3-vessel disease (P < 0.05). <em>Nesfatin</em>-<em>1</em> levels significantly correlated with the number of > 50% stenotic coronary segments (r = 0.<em>1</em>4, P < 0.02). In multivariate analysis, plasma <em>nesfatin</em>-<em>1</em> levels were a significant factor for CAD independent of atherosclerotic risk factors. The odds ratio for CAD was <em>1</em>.7<em>1</em> (95% CI <em>1</em>.0<em>1</em>-2.9<em>1</em>) for high <em>nesfatin</em>-<em>1</em> level of > 0.<em>1</em>9 ng/mL (P < 0.05). Thus, plasma <em>nesfatin</em>-<em>1</em> levels were found to be high in patients with CAD and were associated with CAD independent of atherosclerotic risk factors, suggesting that high <em>nesfatin</em>-<em>1</em> levels in patients with CAD may play a role in the development of coronary atherosclerosis.

This study investigated the cardiovascular effects of <em>nesfatin</em>-<em>1</em> in normotensive rats and animals subjected to hypotensive hemorrhage. Hemorrhagic hypotension was induced by withdrawal 2 mL blood/<em>1</em>00 g body weight over a period of <em>1</em>0 min. Acute hemorrhage led to a severe and long-lasting decrease in mean arterial pressure (MAP) and heart rate (HR). Intracerebroventricularly (i.c.v.) administered <em>nesfatin</em>-<em>1</em> (<em>1</em>00 pmol) increased MAP in both normotensive and hemorrhaged rats. <em>Nesfatin</em>-<em>1</em> also caused bradycardia in normotensive and tachycardia in hemorrhaged rats. Centrally injected <em>nesfatin</em>-<em>1</em> (<em>1</em>00 pmol, i.c.v.) also increased plasma catecholamine, vasopressin and renin concentrations in control animals and potentiated the rise in all three cardiovascular mediators produced by hemorrhage. These findings indicate that centrally administered <em>nesfatin</em>-<em>1</em> causes a pressor response in conscious normotensive and hemorrhaged rats and suggest that enhanced sympathetic activity and elevated vasopressin and renin concentrations mediate the cardiovascular effects of the peptide.

BACKGROUND

Nucleobindin2 (NUCB2)/<em>nesfatin</em>-<em>1</em> plays a well-established role in homeostatic functions associated with food intake and stress integration.

OBJECTIVE

This review focusses on NUCB2/<em>nesfatin</em>-<em>1</em>'s central effects on gastrointestinal functions and will summarize the effects on food intake, motility and secretion with focus on the upper gastrointestinal tract.

RESULTS

We will highlight the stressors that influence brain NUCB2/<em>nesfatin</em>-<em>1</em> expression and discuss functional implications. In addition to traditional acute psychological and physical stressors such as restraint stress and abdominal surgery we will look at immunological, visceral and metabolic stressors as well as a chronic combination stress model that have been shown to affect NUCB2/<em>nesfatin</em>-<em>1</em> signaling and describe associated functional consequences.

Non-alcoholic fatty liver disease (NAFLD) is the most common cause of chronic liver disease, resulting not only in liver dysfunction, glucose and lipid metabolism disorder, but also in neuropsychiatric damage. In the present study, a NAFLD rat model was established via feeding of a high-fat diet, and behaviour was observed via the open field test (OFT), the sucrose preference test (SPT), the elevated plus maze (EPM), the forced swimming test (FST) and the Morris water maze (MWM). The plasma concentrations of alanine aminotransferase (ALT), glucose, free fatty acid (FFA), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) were detected using chemiluminescence technique. The plasma levels of <em>nesfatin</em>-<em>1</em>, leptin and insulin were measured via enzyme-linked immunosorbent assay, and the protein expressions of p-glycogen synthase kinase-3β (GSK-3β), GSK-3β, p-β-catenin, β-catenin, cyclinD and copine 6 in the hippocampus and prefrontal cortex (PFC) were detected using western blotting. After 4 consecutive weeks of feeding with a high-fat diet, the rats showed obesity; increased plasma concentrations of ALT, glucose, FFA, TC, TG, HDL-C and LDL-C; decreased plasma levels of leptin and insulin; and inflammation and mild hepatocyte steatosis in the liver. Although there was no significant difference between groups with regard to performance in the OFT, EPM or FST, the NAFLD rats showed a decreased sucrose preference index in the SPT and impaired learning and memory in the MWM task. Moreover, the present study provides the first evidence of an increased plasma <em>nesfatin</em>-<em>1</em> concentration in NAFLD rats, which was significantly correlated with plasma lipid concentrations and behavioural performance. Furthermore, copine 6 and p-β-catenin protein expression decreased and p-GSK-3β increased in the hippocampus and PFC of NAFLD rats. These results suggest that consuming of a high-fat diet for 4 consecutive weeks could successfully induce a NAFLD rat model. More importantly, these results provide the first evidence that impaired learning and memory in NAFLD rats was, at least partly, associated with increased plasma <em>nesfatin</em>-<em>1</em> concentration and decreased copine 6 expression in the hippocampus and PFC.

Since the discovery of <em>Nesfatin</em>-<em>1</em> in 2006, intensive research was finalized to further and deeper investigate the precise physiological functions of the peptide at both central and peripheral levels, rapidly enriching the knowledge regarding this intriguing molecule. <em>Nesfatin</em>-<em>1</em> is a hypothalamic peptide generated via the post-translational processing of its precursor Nucleobindin 2, a protein supposed to play a role in many biological processes thanks to its ability to bind calcium and to interact with different intracellular proteins. <em>Nesfatin</em>-<em>1</em> is mainly known for its anorexic properties, but it also controls water intake and glucose homeostasis. Recent experimental evidences describe the peptide as a possible direct/indirect orchestrator of central and peripheral cardiovascular control. A specific <em>Nesfatin</em>-<em>1</em> receptor still remains to be identified although numerous studies suggest that the peptide activates extra- and intracellular regulatory pathways by involving several putative binding sites. The present paper was designed to systematically review the latest findings about <em>Nesfatin</em>-<em>1</em>, focusing on its cardiovascular regulatory properties under normal and physiopathological conditions. The hope is to provide the conceptual basis to consider <em>Nesfatin</em>-<em>1</em> not only as a pleiotropic neuroendocrine molecule, but also as a homeostatic modulator of the cardiovascular function and with a crucial role in cardiovascular diseases.

<em>Nesfatin</em>-<em>1</em>, a recently discovered neuropeptide involved in satiety. Recent studies have revealed that central <em>nesfatin</em>-<em>1</em> inhibits gastric emptying and gastric acid secretion, though the mechanisms involved in these processes are not known. We aim to explore the effects of <em>nesfatin</em>-<em>1</em> on a population of gastric distension (GD)-sensitive neurons in the lateral hypothalamus (LHA), gastric motility, and gastric secretion and the role for an arcuate nucleus (Arc)-LHA neural pathway in these processes. Single unit extracellular discharge recordings were made in of LHA. Further, gastric motility and gastric secretion in rats were monitored. Retrograde tracing and fluorescent immunohistochemical staining were used to explore <em>nesfatin</em>-<em>1</em> neuron projection. The results revealed that administration of <em>nesfatin</em>-<em>1</em> to the LHA or electric stimulation of the Arc could alter the neuronal activity of melanin-concentrating hormone (MCH)-responsive, GD-responsive neurons in LHA, which could be blocked by pretreatment with MCH receptor-<em>1</em> antagonist PMC-388<em>1</em>-PI or weakened by pretreatment of a <em>nesfatin</em>-<em>1</em> antibody in LHA. Administration of <em>nesfatin</em>-<em>1</em> into LHA could inhibit gastric motility and gastric secretion, and these effects could be enhanced by administration of PMC-388<em>1</em>-PI. Electrical stimulation of Arc promoted the gastric motility and gastric secretion. <em>Nesfatin</em>-<em>1</em> antibody or PMC-388<em>1</em>-PI pretreatment to LHA had no effect on Arc stimulation-induced gastric motility, but these pretreatments did alter Arc stimulation-induced effects on gastric secretion. Our findings suggest that <em>nesfatin</em>-<em>1</em> signaling in LHA participates in the regulation of efferent information from the gastrointestinal tract and gastric secretion which also involve MCH signaling. Further, they show that a <em>nesfatin</em>-<em>1</em>-positive Arc to LHA pathway is critical for these effects.

OBJECTIVE

We aimed to investigate serum <em>nesfatin</em>-<em>1</em> level in girls with premature thelarche (PT) and its relationship with anthropometric parameters and leptin, which are involved in the initiation of pubertal process.

UNASSIGNED

Non-obese girls who presented with the complaint of early (2-8 years) and isolated breast development were included in the study. The control group consisted of age-matched healthy prepubertal girls. Auxological measurements were performed in all subjects. Gonadotropin-releasing hormone (GnRH) stimulation test and bone age assessment were conducted in subjects with early breast development. Girls with a bone age/chronologic age ratio (<em>1</em>.2 and a peak luteinizing hormone (LH) response to GnRH stimulation <5 mIU/L were included in the PT group.

RESULTS

The study included 22 non-obese girls with PT and 24 healthy prepubertal controls. Body mass index (BMI), BMI-standard deviation score (SDS) and height SDS were similar between the groups (p>> 0.05). Serum leptin and <em>nesfatin</em>-<em>1</em> levels were found significantly higher in the PT group compared to controls (p < 0.05). No correlation was detected between <em>nesfatin</em>-<em>1</em> and basal LH, basal follicle stimulating hormone (FSH), stimulated peak LH, peak FSH, leptin levels and anthropometric parameters in the PT group (p>> 0.05).

CONCLUSIONS

Results of the present study showed that serum <em>nesfatin</em>-<em>1</em> and leptin levels are significantly higher in girls with PT than in prepubertal controls. This finding suggests that similar to leptin, <em>nesfatin</em>-<em>1</em> may also have a central or peripheral role in the initiation of pubertal process and may be related to PT pathogenesis.

OBJECTIVE

The purpose of our investigative work has been to determine whether there can be therapeutic roles in the administration of sildenafil citrate, heparin and several neuropeptides on an animal model where gastric ulcers were induced with acetic acid, and to compare their efficacy.

METHODS

The animals were divided into <em>1</em>3 groups, with 4 animals in each. Gastric ulcers was induced in the animals of <em>1</em>2 groups with one untreated group being left as the control (Group I - control; given normal saline (NS)). The other groups were: Group II (ulcer+NS); Group III (5mg/kg sildenafil citrate, low dose); Group IV (<em>1</em>0mg/kg sildenafil citrate, high dose); Group V (0.6mg/kg heparin, low dose); Group VI (6mg/kg heparin, high dose); Group VII (20nmol/kg des-acyl ghrelin); Group VIII (40nmol/kg des-acyl ghrelin); Group IX (4nmol/kg acyl ghrelin); Group X (8nmol/kg acly ghrelin); Group XI (20pmol/kg <em>Nesfatin</em>-<em>1</em>); Group XII (<em>1</em>5nmol/kg Obestatin) and Group XIII (5nmol/kg Neuropeptide Y). Gastric neuropeptide expression was measured using an immunohistochemical method, and the amount in circulation was detected using ELISA. To compare with no treatment, the controls and other treatment groups, we recorded loss of the surface epithelium of the stomach, erosion, bleeding and inflammatory cell infiltration in the upper halves of the gastric glands.

RESULTS

The muscularis and the layers beneath it were, however, apparently normal. The gastric mucosa healed with little or no inflammation when sildenafil citrate, low dose heparin, ghrelin, NUCB2/<em>Nesfatin</em>-<em>1</em>, obestatin, Neuropeptide Y were administered.

CONCLUSIONS

Overall the data indicate that low dose heparin, and especially sildenafil citrate and neuropeptides, can be used clinically as an alternative approach in the treatment of the gastric ulcer.

Phoenixin is a novel neuropeptide initially associated with reproductive functions, but subsequently also with feeding behavior. <em>Nesfatin</em>-<em>1</em> is also involved in the regulation of food intake and has been shown to largely colocalize with phoenixin in the rat brain; however, a functional link is missing so far. The current study investigated whether phoenixin activates <em>nesfatin</em>-<em>1</em> immunoreactive nuclei in the rat brain. Male Sprague Dawley rats chronically equipped with an intracerebroventricular cannula were injected with vehicle (5 µl ddH₂O) or phoenixin (<em>1</em>.7 nmol in 5 µl ddH₂O, n = 5-6 group). Behavior was assessed manually and c-Fos as well as <em>nesfatin</em>-<em>1</em> immunoreactivity using immunohistochemistry. Phoenixin significantly increased feeding and drinking behavior as well as locomotor activity compared to vehicle (p < 0.0<em>1</em>). Moreover, phoenixin injected intracerebroventricularly (icv) activated several nuclei throughout the rat brain as assessed using c-Fos; the number of c-Fos/<em>nesfatin</em>-<em>1</em> immunoreactive neurons was increased in the lateral septal nucleus (4-fold), supraoptic nucleus (<em>1</em>07-fold), paraventricular nucleus (6-fold) and the nucleus of the solitary tract (<em>1</em>8-fold) compared to vehicle (p < 0.05). In summary, phoenixin activates several <em>nesfatin</em>-<em>1</em> immunoreactive nuclei in the rat brain. This activation may play a role in the modulation of food intake.

The hypothalamus regulates a number of autonomic functions essential for homeostasis; therefore, investigations concerning hypothalamic neuropeptides and their functions and distribution are of great importance in contemporary neuroscience. Recently, novel regulatory factors expressed in the hypothalamus have been discovered, of which <em>nesfatin</em>-<em>1</em> and phoenixin (PNX), show intriguing similarities in their brain distributions. There are currently few studies characterizing PNX expression, so it is imperative to accurately trace its localization, with particular attention to the hypothalamic nuclei and <em>nesfatin</em>-<em>1</em> co-expression. Using fluorescence and classical immunohistochemical stainings on adult rat brain, we visualized the potential co-expression of <em>nesfatin</em>-<em>1</em> and PNX immunoreactive cells. We have demonstrated a distinct PNX-immunoreactivity in 2<em>1</em>-32% of cells in the arcuate nucleus, paraventricular nucleus, ventromedial and lateral hypothalamus. <em>Nesfatin</em>-<em>1</em> expression reached 45-68% of all neurons in the same sites, while co-expression was strikingly seen in the vast majority (70-86%) of PNX-immunoreactive neurons in the rat hypothalamic nuclei. Our results demonstrate for the first time, a wide distribution of PNX in the hypothalamus which could implicate a potential functional relationship with <em>nesfatin</em>-<em>1</em>, possibly in the regulation of the hypothalamic-pituitary-gonadal axis or other autonomic functions, which require further study.

Phoenixin is a recently discovered peptide, which has been associated with reproduction, anxiety and food intake. Based on a considerable co-localization it has been linked to <em>nesfatin</em>-<em>1</em>, with a possible antagonistic mode of action. Since <em>nesfatin</em>-<em>1</em> is known to play a role in anxiety and the response to stress, this study aims to investigate the effects of a well-established psychological stress model, restraint stress, on phoenixin-expressing brain nuclei and phoenixin expression in rats. Male Sprague-Dawley rats were subjected to restraint stress (n = 8) or left undisturbed (control, n = 6) and the brains processed for c-Fos- and phoenixin immunohistochemistry. The number of c-Fos expressing cells was counted and phoenixin expression assessed semiquantitatively. Restraint stress significantly increased c-Fos expression in the dorsal motor nucleus of vagus nerve (DMN, 52-fold, p < 0.00<em>1</em>), raphe pallidus (RPa, <em>1</em>5-fold, p < 0.00<em>1</em>), medial part of the nucleus of the solitary tract (mNTS, <em>1</em>6-fold, p < 0.00<em>1</em>), central amygdaloid nucleus, medial division (CeM, 9-fold, p = 0.0<em>1</em>), supraoptic nucleus (SON, 9-fold, p < 0.00<em>1</em>) and the arcuate nucleus (Arc, 2.5-fold, p < 0.03) compared to control animals. Also phoenixin expression significantly increased in the DMN (<em>1</em>7-fold, p < 0.00<em>1</em>), RPa (2-fold, p < 0.00<em>1</em>) and mNTS (<em>1</em>.6-fold, p < 0.00<em>1</em>) with positive correlations between c-Fos and phoenixin (r = 0.74-0.85; p < 0.0<em>1</em>) in these nuclei. This pattern of activation suggests an involvement of phoenixin in response to restraint stress. Whether phoenixin mediates stress effects or is activated in a counterbalancing fashion will have to be further investigated.

Keywords: Brain-gut; Immobilization; Psychological; Stress; c-Fos.

Activity-based anorexia (ABA) is an established animal model for the eating disorder anorexia nervosa (AN). The pathophysiology of AN and the involvement of food intake-regulatory peptides is still poorly understood. <em>Nesfatin</em>-<em>1</em>, an anorexigenic peptide also involved in the mediation of stress, anxiety and depression might be a likely candidate involved in the pathogenesis of AN. Therefore, activation of <em>nesfatin</em>-<em>1</em> immunoreactive (ir) brain nuclei was investigated under conditions of ABA. Female Sprague-Dawley rats were used and divided into four groups (n=6/group): activity-based anorexia (ABA), restricted feeding (RF), activity (AC) and ad libitum fed (AL). After the 2<em>1</em>-day experimental period and development of ABA, brains were processed for c-Fos/<em>nesfatin</em>-<em>1</em> double labeling immunohistochemistry. ABA increased the number of <em>nesfatin</em>-<em>1</em> immunopositive neurons in the paraventricular nucleus, arcuate nucleus, dorsomedial hypothalamic nucleus, locus coeruleus and in the rostral part of the nucleus of the solitary tract compared to AL and AC groups (p<0.05) but not to RF rats (p>0.05). Moreover, we observed significantly more c-Fos and <em>nesfatin</em>-<em>1</em> ir double-labeled cells in ABA rats compared to RF, AL and AC in the supraoptic nucleus (p<0.05) and compared to AL and AC in the paraventricular nucleus, arcuate nucleus, dorsomedial hypothalamic nucleus, dorsal raphe nucleus and the rostral raphe pallidus (p<0.05). Since <em>nesfatin</em>-<em>1</em> plays a role in the inhibition of food intake and the response to stress, we hypothesize that the observed changes of brain <em>nesfatin</em>-<em>1</em> might play a role in the pathophysiology and symptomatology under conditions of ABA and potentially also in patients with AN.

The aim of the present study was to explore central and peripheral host responses to an anorexia-cachexia producing tumor. We focused on neuroendocrine anorexigenic signals in the hypothalamus, brainstem, pituitary and from the tumor per se. Expression of mRNA for corticotropin-releasing hormone (CRH), cocaine- and amphetamine-regulated transcript (CART), <em>nesfatin</em>-<em>1</em>, thyrotropin (TSH) and the TSH receptor were explored. In addition, we examined changes in plasma TSH, CART peptides (CARTp) and serum amyloid P component (SAP). C57BL/6 mice were implanted with MCG<em>1</em>0<em>1</em> tumors or sham-treated. A sham-implanted, pair‑fed (PF) group was included to delineate between primary tumor and secondary effects from reduced feeding. Food intake and body weight were measured daily. mRNA levels from microdissected mouse brain samples were assayed using qPCR, and plasma levels were determined using ELISA. MCG<em>1</em>0<em>1</em> tumors expectedly induced anorexia and loss of body weight. Tumor-bearing (TB) mice exhibited an increase in <em>nesfatin</em>-<em>1</em> mRNA as well as a decrease in CART mRNA in the paraventricular area (PVN). The CART mRNA response was secondary to reduced caloric intake whereas <em>nesfatin</em>-<em>1</em> mRNA appeared to be tumor-specifically induced. In the pituitary, CART and TSH mRNA were upregulated in the TB and PF animals compared to the freely fed controls. Plasma levels for CARTp were significantly elevated in TB but not PF mice whereas levels of TSH were unaffected. The plasma CARTp response was correlated to the degree of inflammation represented by SAP. The increase in <em>nesfatin</em>-<em>1</em> mRNA in the PVN highlights <em>nesfatin</em>-<em>1</em> as a plausible candidate for causing tumor-induced anorexia. CART mRNA expression in the PVN is likely an adaptation to reduced caloric intake secondary to a cancer anorexia-cachexia syndrome (CACS)‑inducing tumor. The MCG<em>1</em>0<em>1</em> tumor did not express CART mRNA, thus the elevation of plasma CARTp is host derived and likely driven by inflammation.

NUCB2/<em>nesfatin</em> and its proteolytically cleaved product <em>nesfatin</em>-<em>1</em> are recently discovered anorexigenic hypothalamic neuroproteins involved in energy homeostasis. It is expressed both centrally and in peripheral tissues, and appears to have potent metabolic actions. NUCB2/<em>nesfatin</em> neurons are activated in response to stress. Central <em>nesfatin</em>-<em>1</em> administration elevates circulating ACTH and corticosterone levels. Bilateral adrenalectomy increased NUCB2/<em>nesfatin</em> mRNA levels in rat paraventricular nuclei. To date, studies have not assessed the effects of <em>nesfatin</em>-<em>1</em> stimulation on human adrenocortical cells. Therefore, we investigated the expression and effects of <em>nesfatin</em>-<em>1</em> in a human adrenocortical cell model (H295R). Our findings demonstrate that NUCB2 and <em>nesfatin</em>-<em>1</em> are expressed in human adrenal gland and human adrenocortical cells (H295R). Stimulation with <em>nesfatin</em>-<em>1</em> inhibits the growth of H295R cells and promotes apoptosis, potentially via the involvement of Bax, BCL-XL and BCL-2 genes as well as ERK<em>1</em>/2, p38 and JNK<em>1</em>/2 signalling cascades. This has implications for understanding the role of NUCB2/<em>nesfatin</em> in adrenal zonal development. NUCB2/<em>nesfatin</em> may also be a therapeutic target for adrenal cancer. However, further studies using in vivo models are needed to clarify these concepts.

<b>Background/objective:</b> This study aimed to investigate the impacts of a <em>1</em>2-week training of the aerobic exercise (AE), resistance exercise (RE), and combined exercise (CE) on the serum levels of <em>nesfatin</em>-<em>1</em>, irisin-<em>1</em> and some other metabolic and anthropometric indices in overweight women with metabolic syndrome. <b>Methods:</b> Sixty overweight women with metabolic syndrome were assigned equally into four groups: aerobic exercise (AE, <i>n</i> = <em>1</em>5), resistance exercise (RE, <i>n</i> = <em>1</em>5), combined exercise (CE, <i>n</i> = <em>1</em>5), and control (<i>n</i> = <em>1</em>5). All groups underwent <em>1</em>2 weeks of intervention. The study variables were measured before and 24 h after the intervention period. <b>Results:</b> Twelve weeks of training resulted in an increase of irisin-<em>1</em> in the AE and CE groups and <em>nesfatin</em>-<em>1</em> in all the intervention groups. As expected, all the trained groups exhibited a positive alteration in anthropometric indices and lipid profile in comparison with the control group. Besides, compared with the control group, insulin resistance (based on the homeostatic model assessment) in AE (<i>p</i> = 0.022), RE (<i>p</i> = 0.032), and CE (<i>p</i> < 0.00<em>1</em>) groups were reduced significantly. According to the observed changes in the measured indices, serum irisin-<em>1</em> was significantly correlated with body weight, BMI, body fat percentage, fasting insulin, and HOMA-IR. However, with regard to <em>nesfatin</em>-<em>1</em>, only a negative correlation was observed with body fat percentage and LDL-cholesterol. <b>Conclusions:</b> The <em>1</em>2-week systematic training program changed circulating irisin-<em>1</em> and <em>nesfatin</em>-<em>1</em>. Also, change in the serum irisin-<em>1</em> and <em>nesfatin</em>-<em>1</em> were correlated with the change in glycemic and anthropometric indices in addition to LDL-cholesterol. Also, exercise training significantly reduced fasting insulin and HOMA-IR in all the intervention groups. <b>RCT Registration Code:</b> IRCT20<em>1</em>8080604072<em>1</em>N2.

<strong class="sub-title"> Keywords: </strong> HOMA-IR; aerobic; exercise program; insulin; irisin; metabolic syndrome; <em>nesfatin</em>-<em>1</em>; resistance.

Testicular torsion causes ischemia-reperfusion injury and an increased risk of infertility. <em>Nesfatin</em>-<em>1</em> is a novel peptide with antioxidant, anti-inflammatory and anti-apoptotic properties. In the present study, we aimed to investigate the putative beneficial effects of <em>nesfatin</em>-<em>1</em> on oxidative injury and impaired testicular function induced by testis torsion. Under anesthesia, male Sprague-Dawley rats (<em>1</em>80-230 g; n = 24) had sham-operation or they underwent testicular torsion by rotating the left testis 720° and fixing it for 2 h, followed by a 2-h detorsion. Rats in each group were treated intraperitoneally with either <em>nesfatin</em>-<em>1</em> (0.3 μg/kg) or saline prior to the torsion or sham-torsion. At the end of the 4-h experimental period, tissue samples were removed for evaluation of spermatozoa, molecular and histochemical analyses. In saline-treated torsion/detorsion group, a high percentage of abnormal spermatozoa with head defects was observed, which was abolished in <em>nesfatin</em>-<em>1</em>-treated torsion/detorsion group. The levels of 8-OHdG, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, caspase-3 were increased in the saline-treated torsion/detorsion group as compared to sham-operated group, while <em>nesfatin</em>-<em>1</em> pre-treatment significantly decreased the expressions of the pro-inflammatory cytokines, depressed apoptosis, and also reduced the tubular degeneration. In addition, <em>nesfatin</em>-<em>1</em> in torsion/detorsion group elevated expressions of transforming growth factor (TGF)-beta and reduced expressions of protein kinase B (AKT) and cAMP response element binding protein (CREB) in the testis tissue. The present findings show that <em>nesfatin</em>-<em>1</em>, by regulating AKT and CREB signaling pathways and pro-inflammatory/anti-inflammatory cytokine balance, preserves the spermatogenic cells and ameliorates torsion-detorsion-induced tubular degeneration.

Ghrelin, and <em>nesfatin</em>-<em>1</em> (encoded by nucleobindin2/nucb2) are two metabolic peptides with multiple biological effects in vertebrates. While sex steroids are known to regulate endogenous ghrelin and NUCB2 in mammals, such actions by steroids in fish remain unknown. This study aimed to determine whether estradiol (E2) and testosterone (T) affects the expression of preproghrelin, ghrelin/growth hormone secretagogue receptor (GHS-R), ghrelin O-acyl transferase (GOAT) and NUCB2 in goldfish (Carassius auratus). First, a dose-response assay was performed in which fish were intraperitoneally (ip) implanted with pellets containing 25, 50 or <em>1</em>00 μg/g body weight (BW) of E2 or T. It was found that sex steroids (<em>1</em>00 μg/g BW) administered for 2.5 days achieved the highest E2 or T in circulation. In a second experiment, fish were ip implanted with pellets containing <em>1</em>00 μg/g BW of E2, T or without hormone (control). RT-qPCR analyses at 2.5 days post-administration show that gut preproghrelin and GOAT expression was upregulated by both E2 and T treatments, while the same effect was observed for GHS-R only in the pituitary. Both treatments also reduced hypothalamic preproghrelin mRNA expression. NUCB2 expression was increased in the forebrain of T treated group and reduced in the gut and pituitary under both treatments. These results show for the first time a modulation of preproghrelin and nucb2/<em>nesfatin</em>-<em>1</em> by sex steroids in fish. The interaction between sex steroids and genes implicated in both metabolism and reproduction might help meeting the reproduction dependent energy demands in fish.

OBJECTIVE

<em>Nesfatin</em>-<em>1</em> identified neuroendocrine peptide is involved in regulation of homeostasis via modulation of metabolism, energy homeostasis and food intake. We aimed to investigate the associations of circulating <em>nesfatin</em>-<em>1</em> level with food intake, body composition and resting metabolic rate (RMR) and also examine the correlation between circulating peroxisome proliferator-activated receptor gamma (PPARγ) and <em>nesfatin</em>-<em>1</em> levels in obese and morbid obese subjects.

METHODS

A total of 96 obese subjects (including <em>1</em>8 morbid obese subjects) were participated in the current cross-sectional study. We assessed the body composition with the use of Body Composition Analyzer. RMR was measured by means of the MetaCheck™, an instrument designed to measure RMR using indirect calorimetry. All baseline blood samples were obtained following an overnight fasting. Plasma concentrations of <em>nesfatin</em>-<em>1</em> and circulating PPARγ were measured with the use of an ELISA method. Statistical analyses were performed using SPSS.

RESULTS

We found significant associations between fat percent and circulating <em>nesfatin</em>-<em>1</em> in obese and morbid obese subjects. There was main association between circulating <em>nesfatin</em>-<em>1</em> and PPARγ concentration in obese subjects and it was more strong association in morbid obese participants. There was marginally significant differences between percent predicted RMR between different categorized <em>nesfatin</em>-<em>1</em> levels. There were also higher intakes of calorie, carbohydrate and protein in obese group who had lower concentration of <em>nesfatin</em>-<em>1</em>.

CONCLUSIONS

Our data indicated the fat percent as main determinant factor in circulating <em>nesfatin</em>-<em>1</em> level. It appears <em>nesfatin</em>-<em>1</em> and PPARγ might be concurrently involved in adipogenesis pathway.

<AbstractText>Nucleobinding-2 (NUCB2) or <em>nesfatin</em>-<em>1</em>, a newly identified anorexigenic peptide, has antioxidant, anti-inflammatory, and anti-apoptotic properties. Brain ischemiareperfusion induces irreversible damages, especially in the hippocampus area. However, the therapeutic effects of NUCB2 have not been well investigated in cerebral ischemia. This study was designed for the first time to investigate the protective effects of NUCB2/<em>Nesfatin</em>-<em>1</em> on the expression of apoptosis-related proteins and reactive astrogliosis level in the CA<em>1</em> area of hippocampus in an experimental model of transient global cerebral ischemia.</AbstractText><AbstractText>The male Wistar rats were randomly allocated into 4 groups (sham, NUCB2, ischemia-reperfusion, and ischemia-reperfusion+NUCB2<em>1</em>) (n =7). The model of cerebral ischemia was prepared by common carotid arteries occlusion for 20 minutes. <em>Nesfatin</em>-<em>1</em> (20 μg/kg) and saline (as a vehicle) were injected (intraperitoneally) at the beginning of the reperfusion period. The assessment of the protein expression levels was performed by immunofluorescence and immunohistochemical staining.</AbstractText><AbstractText>NUCB2 significantly reduced the Bax and GFAP protein levels in the CA<em>1</em> area after ischemia (P<0.05). Also, NUCB2 increased Bcl-2 protein level (P<0.05). NUCB2 exerted protective effects against ischemic injury by the inhibition of astrocytes activation as an inflammatory response and decreased neuronal cell apoptosis.</AbstractText><AbstractText>The present study provides the possible neuroprotective view of <em>nesfatin</em>-<em>1</em> in the treatment of ischemia injury model in rat hippocampus.</AbstractText>

Recent studies have revealed <em>nesfatin</em>-<em>1</em> as a hypothalamic neuropeptide, regulating food intake, energy expenditure and reproduction primarily by acting on the hypothalamic-pituitary-gonadal axis. <em>Nesfatin</em>-<em>1</em> is also localized in several peripheral tissues including testes. However, functional significance of <em>nesfatin</em>-<em>1</em> in testicular activities is not yet well documented in mammals. Therefore, this study was aimed to elucidate the direct effects of <em>nesfatin</em>-<em>1</em> on testicular markers for steroid productions, spermatogenesis, metabolic changes and oxidative stress. The results revealed the expression of both protein and mRNA of <em>nesfatin</em>-<em>1</em> in the testes of adult mice. The testes treated in vitro with <em>nesfatin</em>-<em>1</em> showed significant increase in testosterone production, which correlated significantly with increased expression of steroidogenic markers and insulin receptor proteins in the testes. Furthermore, the in vitro treatment with <em>nesfatin</em>-<em>1</em> showed stimulatory effects on spermatogenesis by promoting cell proliferation (PCNA) and survival (Bcl2), while inhibiting apoptosis (caspase-3) in the testes. The <em>nesfatin</em>-<em>1</em> treatment in vitro further increased the expression of insulin receptor and GLUT8 proteins, in parallel with increase in the intra-testicular transport of glucose and production of lactate. This <em>nesfatin</em>-<em>1</em> induced enhanced transport of energy substrate (glucose and lactate) may be responsible for promoting spermatogenesis and steroidogenesis. <em>Nesfatin</em>-<em>1</em> significantly reduced oxidative stress and nitric oxide, which may also be responsible for stimulatory effects on testicular activities. The present finding suggests that <em>nesfatin</em>-<em>1</em> acts via paracrine manner to increase sperm count and fertility, thus promoting the testicular function.