OBJECTIVE

To investigate the change of plasma <em>nesfatin</em>-<em>1</em> concentration in a nonalcoholic fatty liver disease rat model induced by high-fat diet, and explore its effect on the dysfunction of glucose and lipid metabolism.

METHODS

The nonalcoholic fatty liver disease rat model was established through introduction of a high-fat diet, and four weeks later, the intraperitoneal glucose tolerance test was conducted. Serum concentrations of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), direct bilirubin (DBIL), indirect bilirubin (IBIL), 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 (INS) were measured via enzyme-linked immunosorbent assay (ELISA), the histological changes of the liver was observed via HE staining, and the protein expressions of beta-catenin, p-beta-catenin and cyclin D<em>1</em> in the liver were detected using western blot and compared with beta-actin.

RESULTS

The bodyweight, liver weight, liver index, and area under the curve of the intraperitoneal glucose tolerance test were all higher in the model rats than those in the controls. Compared with the control, serum concentrations of ALT, TBIL, IBIL, TC and LDL-C in the model rats were all increased. The plasma <em>nesfatin</em>-<em>1</em> level was increased in model rats while the plasma concentrations of leptin and insulin were decreased, and a negative correlation was found between the plasma concentration of <em>nesfatin</em>-<em>1</em> and leptin. Inflammation and hepatocyte steatosis were detected in the livers of model rats, and the protein expression of cyclinD<em>1</em> was upregulated while the phosphorylation of beta-catenin was decreased in the livers of the model rats.

CONCLUSIONS

Post-creation of nonalcoholic fatty liver disease rat models through high fat diets, changes were observed in plasma <em>nesfatin</em>-<em>1</em> concentration, perhaps a vital part of glucose and lipid metabolism dysfunction.

OBJECTIVE

We aimed to observe the effects of apelin supplementation on the plasma levels of <em>nesfatin</em>-<em>1</em> in DOCA-salt hypertensive and normal rats.

METHODS

For this purpose, 28 young Wistar albino male rats were divided into four groups; Control (C), Control + Apelin (C+A), Hypertension (HT) and Hypertension + Apelin (HT+A). Hypertension was induced by injection of DOCA-salt (25 mg/kg, s.c.) twice weekly, 4 weeks, whereas intraperitoneal apelin was administered (200 µg.kg-<em>1</em>) for <em>1</em>7 days. Plasma <em>nesfatin</em>-<em>1</em> and apelin levels were measured with ELISA. Systolic blood pressure was monitored using a tail cuff system. The relationships between plasma <em>nesfatin</em> levels and blood pressure were assessed.

RESULTS

Plasma <em>nesfatin</em>-<em>1</em> levels was found lower in control animals compared to C+A, HT and HT+A groups (p = 0.002, p = 0.026 and p = 0.0<em>1</em><em>1</em>, respectively). Systolic blood pressures were similar in the C and C+A groups, but systolic blood pressures of the HT and HT+A groups was found significantly higher than the C and C+A groups.

CONCLUSIONS

In conclusion, apelin administration induced an increment of <em>nesfatin</em>-<em>1</em> in normal rats and plasma levels of <em>nesfatin</em>-<em>1</em> increase in DOCA-salt hypertension rats. But apelin addition in hypertension did not cause an extra increase in <em>nesfatin</em>-<em>1</em> levels. This is the first report to investigate the effect of apelin administration on plasma <em>nesfatin</em> levels of normal and hypertensive rats (Fig. 2, Ref. 44).

<em>Nesfatin</em>-<em>1</em>, a centrally acting anorexigenic peptide, is produced in several brain areas involved in metabolic processes and has been implicated in the control of ingestive behaviours and cardiovascular functions. The present study aimed to determine whether the subfornical organ (SFO), a central nervous system (CNS) site that has been extensively implicated in the regulation of appetite and thirst, may represent a potential site for central actions of <em>nesfatin</em>-<em>1</em>. We first used the reverse transcriptase-polymerase chain reaction and were able to confirm the presence of mRNA for the nucleobindin-2 gene in the SFO. We then used whole-cell patch clamp recordings to investigate the influence of <em>nesfatin</em>-<em>1</em> on the membrane potential of dissociated SFO neurones. A total of 80.3% (49 of 6<em>1</em>) of neurones tested showed a response to <em>nesfatin</em>-<em>1</em> (<em>1</em>00 nm, <em>1</em>0 nm and <em>1</em> nm). Of these, 47.5% depolarised, with a mean depolarisation of 8.2 ± 0.9 mV (n = 29) and 32.8% hyperpolarised with a mean hyperpolarisation of -8.9 ± <em>1</em>.2 mV (n = 20). Peak magnitudes were seen at a concentration of <em>1</em> nm <em>nesfatin</em>-<em>1</em>, whereas no effect was observed at <em>1</em>00 pm nesftain-<em>1</em> (n = 3). Furthermore, voltage clamp ramp and step protocols revealed a <em>nesfatin</em>-<em>1</em> induced activation of the delayed rectifier potassium conductance, IK . Pharmacological blockade of this conductance greatly reduced the magnitude and occurrence of the observed hyperpolarisations. The present study thus demonstrates that <em>nesfatin</em>-<em>1</em> has the ability to influence the membrane potential of SFO neurones, and thus identifies the SFO as a potential site at which <em>nesfatin</em>-<em>1</em> may act to regulate ingestive behaviour and cardiovascular control.

<em>Nesfatin</em>-<em>1</em> is an anorexic peptide derived from a precursor, nucleobindin-2 (NUCB2), which is distributed in various organs, coexists with ghrelin in the gastric X/A-like cells and closely relates to an appetite control in rodents and humans. <em>Nesfatin</em>-<em>1</em> may be a significant factor addressing the satiety also in veterinary medicine, however, there are few reports about <em>nesfatin</em>-<em>1</em> in dogs. In the present study, we detected canine NUCB2/<em>nesfatin</em>-<em>1</em> mRNA in various tissues, especially abundant in pancreas, gastrointestinal tracts, testis and cerebellum. We examined circulating <em>nesfatin</em>-<em>1</em> concentrations and NUCB2/<em>nesfatin</em>-<em>1</em> mRNA expressions in upper gastrointestinal tracts (gastric corpus, pyloric antrum and duodenum) in dogs fed on different types of diets. Plasma <em>nesfatin</em>-<em>1</em> concentrations in the dogs were approximately 4 ng/ml and they did not change after feeding through the study, however, NUCB2/<em>nesfatin</em>-<em>1</em> mRNA expressions in pyloric antrum were <em>1</em>.84-fold higher in the dogs fed on a High fiber/High protein diet (P<0.00<em>1</em>), <em>1</em>.48-fold higher in the dogs fed on a High fat/Low protein diet (P<0.05) and <em>1</em>.02-fold higher in the dogs fed on a Low fat/High carbohydrate diet (not significant) comparing to those on a control diet. It was concluded that High fiber/High protein and High fat/Low protein diets increased NUCB2/<em>nesfatin</em>-<em>1</em> production in canine gastrointestinal tracts. These results may set the stage for further investigations of canine NUCB2/<em>nesfatin</em>-<em>1</em>, which may relate to satiety effects in dogs.

Mammalian, or mechanic, target of rapamycin (mTOR) signaling is a crucial factor in the regulation of the energy balance that functions as an energy sensor in the body. The present review explores how the mTOR/S6k intracellular pathway is involved in modulating the production of different signals such as ghrelin and <em>nesfatin</em>-<em>1</em> in the gastrointestinal tract to regulate food intake and body weight. The role of gastric mTOR signaling in different physiological processes was studied in depth through different genetic models that allow the modulation of mTOR signaling in the stomach and specifically in gastric X/A type cells. It has been described that mTOR signaling in X/A-like gastric cells has a relevant role in the regulation of glucose and lipid homeostasis due to its interaction with different organs such as liver and adipose tissue. These findings highlight possible therapeutic strategies, with the gut-brain axis being one of the most promising targets in the treatment of obesity.

<strong class="sub-title"> Keywords: </strong> gastrointestinal tract; ghrelin; homeostasis; mTOR; <em>nesfatin</em>-<em>1</em>; obesity.

BACKGROUND

Bariatric surgery is associated with multiple endocrine and metabolic changes. Irisin and <em>nesfatin</em>-<em>1</em> have recently been described as regulatory peptides involved in obesity-related insulin resistance. Our aim was to analyze the changes of those two molecules observed in patients after bariatric procedures.

METHODS

This prospective study involved 40 patients treated for morbid obesity. Irisin and <em>nesfatin</em>-<em>1</em> were measured before, 6 months and <em>1</em> year after surgical intervention. We also gathered demographic data, information concerning comorbidities, factors related to the surgery and outcomes of bariatric treatment.

RESULTS

Twenty-seven patients completed the study (<em>1</em>5 females). The mean age of the group was 43.5 ± <em>1</em>0.4 years. Six (22.2%) patients were submitted to Laparoscopic Sleeve Gastrectomy and 2<em>1</em> (77.8%) patients were submitted to Laparoscopic Roux-en-Y Gastric Bypass. The participants in our study achieved significant weight loss. The irisin level remained stable in the whole study group during all three measurements included in our study protocol (p = .7<em>1</em>). Our study group presented a reduction of the <em>nesfatin</em>-<em>1</em> level 6 months after bariatric surgery and a slight further decrease after one-year observation, although these changes were also not significant (p = .<em>1</em>7).

CONCLUSIONS

We did not find any significant correlation between changes of irisin or <em>nesfatin</em>-<em>1</em> level and bariatric surgery, as an aid in the regulation of glucose metabolism.

There is growing evidence that obstructive sleep apnoea (OSA) influences the hypothalamic-pituitary-gonadal axis (HPG axis) in men. The aim of the study was to assess the association of <em>nesfatin</em>-<em>1</em> with HPG axis disturbances in OSA. This is a prospective study with consecutive enrolment. It comprises 72 newly diagnosed OSA patients ((AHI: apnoea-hypopnea index) <em>1</em>8 subjects: 5 ≤ AHI < <em>1</em>5; 24: <em>1</em>5 ≤ AHI < 30; 30: AHI ≥ 30) and a control group composed of <em>1</em>9 patients (AHI < 5). All patients underwent polysomnography and fasting blood collection for <em>nesfatin</em>-<em>1</em>, testosterone, luteinising hormone (LH), high-sensitivity C-reactive protein (hsCRP), aspartate transaminase (AST), alanine aminotransferase (ALT), creatinine and glucose. Groups had similar levels of LH, <em>nesfatin</em>-<em>1</em> and testosterone (p = 0.87; p = 0.24; p = 0.08). <em>Nesfatin</em>-<em>1</em> was not correlated to LH (p = 0.7<em>1</em>), testosterone (p = 0.38), AHI (p = 0.34) or the oxygen desaturation index (ODI) (p = 0.69) either in the whole group, or in sub-groups. The study did not reveal any association between the HPG axis and <em>nesfatin</em>-<em>1</em> in OSA adult males. It is possible that <em>nesfatin</em>-<em>1</em> is not a mediator of HPG axis disturbances in adult patients with OSA.

<em>Nesfatin</em>-<em>1</em> is a multifunctional neuropeptide having crucial autonomic roles. It is well known that <em>nesfatin</em>-<em>1</em> collaborates with other central neuromodulatory systems, such as central corticotropin-releasing hormone, melanocortin, oxytocin, and cholinergic systems to show its autonomic effects. Central arachidonic acid cascade plays an important role to provide the homeostasis by exhibiting similar autonomic effects to <em>nesfatin</em>-<em>1</em>. Based on these similarities, the current study was designed to show the effects of intracerebroventricularly (ICV) injected <em>nesfatin</em>-<em>1</em> on the hypothalamic arachidonic acid (AA) cascade. Immunochemistry and western blot approaches demonstrated that ICV administration of <em>nesfatin</em>-<em>1</em> provokes an increase in the hypothalamic cyclooxygenase (COX) -<em>1</em>, -2 and lipoxygenase (LOX) protein expression. Moreover, the microdialysis study demonstrated that centrally injected <em>nesfatin</em>-<em>1</em> increased the posterior hypothalamic extracellular AA products. In conclusion, these findings report that while <em>nesfatin</em>-<em>1</em> is generating its autonomic effects, it also might be using central prostaglandins and leukotrienes by activating central COX and LOX pathways.

<em>Nesfatin</em>-<em>1</em> (NES<em>1</em>) is a potential biomarker found in serum and saliva that indicates hyperpolarization and depolarization in the hypothalamic ventricle nucleus as well as an increase in epileptic conditions. However, real-time investigations have not been carried out to detect changes in the concentration of NES<em>1</em>. In this study, we develop a multiscale pore contained carbon nanofiber-based field-effect transistor (FET) biosensor to detect NES<em>1</em>. The activated multiscale pore contained carbon nanofiber (a-MPCNF) is generated using a single-nozzle co-electrospinning method and a subsequent steam-activation process to obtain a signal transducer and template for immobilization of bioreceptors. The prepared biosensor exhibits a high sensitivity to NES<em>1</em>. It can detect levels as low as 0.<em>1</em> fM of NES<em>1</em>, even in the presence of other interfering biomolecules. Furthermore, the a-MPCNF-based FET sensor has significant potential for practical applications in non-invasive real-time diagnosis, as indicated by its sensing performance in artificial saliva.

Nucleobindin (NUCB)-derived peptides, <em>nesfatin</em>-<em>1</em> (NES-<em>1</em>) and <em>nesfatin</em>-<em>1</em>-like peptide (NLP) have several physiological roles in vertebrates. While NES-<em>1</em> is implicated in stress, whether NUCB<em>1</em>/NLP and NUCB2/NES-<em>1</em> have any effect on proopiomelanocortin (POMC) remains unknown. The main aim of this study was to determine if NES-<em>1</em> and/or NLP affect POMC synthesis in mouse corticotrophs. Immunocytochemistry was employed to target NUCB colocalization with POMC in immortalized mouse tumoral corticotrophs (AtT-20 cells). The ability of NES-<em>1</em> and NLP to modulate POMC mRNA and protein in AtT-20 cells was assessed by qPCR and Western blot, respectively. Moreover, cell-signaling molecules mediating the effect of NES-<em>1</em> and NLP on POMC synthesis in mouse tumoral corticotrophs were studied using pharmacological blockers. Mouse tumoral corticotrophs showed immunoreactivity for both NUCB<em>1</em>/NLP and NUCB2/NES-<em>1</em>. Both NES-<em>1</em> and NLP exerted a stimulatory effect on POMC transcript abundance and protein expression in a dose- and time-dependent manner. This effect was comparable to corticotropin-releasing factor (CRF, positive control) stimulation of POMC. Incubation of mouse tumoral corticotrophs with NES-<em>1</em> or NLP upregulated the phosphorylation of protein kinase A (PKA) and cAMP-response element-binding protein (CREB). The stimulatory effect of these peptides on POMC transcript abundance and protein expression was blocked by the PKA inhibitor, H89, and an adenylate cyclase inhibitor, 2',3'-dideoxyadenosine (DDA). These pharmacological studies indicate that NES-<em>1</em> and NLP act through the cAMP/PKA/CREB cellular pathway to stimulate POMC synthesis. Our results provide molecular evidence to support a stimulatory role for nucleobindin-derived peptides on POMC synthesis from corticotrophs. Collectively, this research indicates that corticotrophs produce NUCBs, and the encoded peptides NES-<em>1</em> and NLP could elicit a direct action to stimulate the pituitary stress hormone. This stimulatory effect is mediated by an uncharacterized G protein-coupled receptor (GPCR) that utilizes the cAMP/PKA/CREB pathway.

<strong class="sub-title"> Keywords: </strong> Corticotrophs; Nesfatin-<em>1</em>; Nesfatin-<em>1</em>-like peptide; Proopiomelanocortin; Stress.

<em>Nesfatin</em>-<em>1</em>, an 82 amino acid peptide cleaved from the N-terminal of its precursor nucleobindin-2 (NUCB2), is emerging as a multifunctional peptide in fish. The present study aimed to determine whether <em>nesfatin</em>-<em>1</em> plays a role in fish somatic growth by modulating the growth hormone (GH)/insulin-like growth factor (IGF) axis, using a representative teleost model, the goldfish (Carassius auratus). The results demonstrated that a single i.p. injection of synthetic goldfish <em>nesfatin</em>-<em>1</em> significantly decreased the expression of hypothalamic pacap (approximately 90%) and pituitary Gh (approximately 90%) mRNAs at <em>1</em>5 minutes post-injection. Serum GH levels were also reduced as a result of <em>nesfatin</em>-<em>1</em> administration, by approximately 45% and 55% at <em>1</em>5 and 30 minutes post-injection, respectively. Likewise, in vitro treatment of goldfish dispersed pituitary cells with <em>nesfatin</em>-<em>1</em> reduced Gh secretion, suggesting that <em>nesfatin</em>-<em>1</em> acts directly on pituitary somatotrophs to inhibit Gh release. Exposure of cultured liver fragments to <em>nesfatin</em>-<em>1</em> (0.<em>1</em>, <em>1</em> and <em>1</em>0 nmol L^-<em>1</em> ) led to a significant reduction in igf-<em>1</em> mRNA at <em>1</em>20 minutes and of igf-II mRNA at 30 and 60 minutes post-incubation. Collectively, these results indicate a suppressive role for <em>nesfatin</em>-<em>1</em> on the goldfish GH/IGF axis. Immunohistochemical studies demonstrated that NUCB2/<em>nesfatin</em>-<em>1</em>-like immunoreactivity, although present in the goldfish pituitary, is not colocalised with GH in goldfish somatotrophs. Thus, <em>nesfatin</em>-<em>1</em> does not appear to act in an autocrine manner to regulate GH secretion. Taken together, this research found that the pituitary gland is an important source of endogenous NUCB2/<em>nesfatin</em>-<em>1</em> and also that <em>nesfatin</em>-<em>1</em> directly suppresses the Gh/IGF axis in goldfish.

<strong class="sub-title"> Keywords: </strong> GH; IGF-I and IGF-II; NUCB2; PACAP; fish; <em>nesfatin</em>-<em>1</em>; pituitary.

During the teleost radiation, extensive development of the direct innervation mode of hypothalamo-pituitary communication was accompanied by loss of the median eminence typical of mammals. Cells secreting follicle-stimulating hormone and luteinizing hormone cells are directly innervated, distinct populations in the anterior pituitary. So far, ∼20 stimulatory and ∼<em>1</em>0 inhibitory neuropeptides, 3 amines, and 3 amino acid neurotransmitters are implicated in the control of reproduction. Positive and negative sex steroid feedback loops operate in both sexes. Gene mutation models in zebrafish and medaka now challenge our general understanding of vertebrate neuropeptidergic control. New reproductive neuropeptides are emerging. These include but are not limited to <em>nesfatin</em> <em>1</em>, neurokinin B, and the secretoneurins. A generalized model for the neuroendocrine control of reproduction is proposed. Hopefully, this will serve as a research framework on diverse species to help explain the evolution of neuroendocrine control and lead to the discovery of new hormones with novel applications. Expected final online publication date for the <i>Annual Review of Animal Biosciences</i>, Volume <em>1</em>0 is February 2022. Please see http://www.annualreviews.org/page/journal/pubdates for revised estimates.

Along with the developing technology in the modern age, physical activity had decreased considerably in children and adolescents alike with a concomittant and rapid increase in the prevalence of childhood obsesity. The purpose of the present study is to measure the levels of serum <em>nesfatin</em>-<em>1</em> and irisin in obese children. The present study was carried out with a total of 62 children, including 32 obese children diagnosed between June 20<em>1</em>7 and October 20<em>1</em>7 and 30 healthy children. Serum <em>nesfatin</em>-<em>1</em>, irisin, SOD, MDA, fasting blood glucose, total cholesterol (TC), triglyceride (TG), HDL-C, LDL-C, aspartate amino transferase (AST), alanine amino transferase (ALT)), blood urea nitrogen (BUN), C-reactive protein (CRP), calcium (Ca), sodium (Na), potassium (P), chromium (Cr), ferritin, and vitamin B_<em>1</em>2 data were collected for each patient. In our study, mean <em>nesfatin</em>-<em>1</em> and SOD values of the obesity group were lower than those of the control group (p <0.05, p <0.00<em>1</em>), whereas irisin and MDA values were higher than those of the control group (p <0.00<em>1</em>). Childhood obesity is still a significant global problem, despite increased social awareness and numerous preventive healthcare interventions. We believe that all the prospective studies to be carried out to evaluate the relationship between obesity-irisin-<em>nesfatin</em>-<em>1</em> triad, will make positive contributions to treatment of obesity.

<strong class="sub-title"> Keywords: </strong> Irisin; <em>nesfatin</em>-<em>1</em>; obesity; oxidative stress.

<em>Nesfatin</em>-<em>1</em>, a pleotropic peptide, was recently implicated in the regulation of anxiety and depression-like behavior in rats. However, the underlying mechanisms remain unclear so far. Thus, this study aimed to investigate the role of endogenous <em>nesfatin</em>-<em>1</em> in the mediation of anxiety and depression-like behavior induced by corticotropin-releasing factor (CRF). Therefore, normal weight male intracerebroventricularly (icv) cannulated Sprague Dawley rats received two consecutive icv injections of anti-<em>nesfatin</em>-<em>1</em> antibody or IgG control antibody followed by CRF or saline, before being exposed to a behavioral test. In the elevated zero maze test, assessing anxiety and explorative behavior, blockade of <em>nesfatin</em>-<em>1</em> using an anti-<em>nesfatin</em>-<em>1</em> antibody under basal conditions increased the number of entries into the open arms compared to control antibody/vehicle (<em>1</em>.6-fold, p < 0.05) and the time in open arms compared to the other groups (p < 0.05). Control antibody/CRF-treated animals tended to spend less time in the open arms compared to control antibody/vehicle (0.7-fold, p = 0.<em>1</em>7), an effect not altered by the <em>nesfatin</em>-<em>1</em> antibody (control antibody/CRF-treated animals vs. <em>nesfatin</em>-<em>1</em> antibody/CRF group, p = <em>1</em>.00). In the novelty-induced hypophagia test, assessing anhedonia as part of depression-like behavior, no significant differences were observed between the four groups for the latency to the first bout, number of bouts and the amount of palatable snack eaten (p > 0.05). In summary, CRF tended to increase anxiety and explorative behavior an effect not altered by blockade of <em>nesfatin</em>-<em>1</em>, whereas no significant effect of CRF on anhedonia was observed. Blockade of endogenous <em>nesfatin</em>-<em>1</em> significantly decreased anxiety-like behavior giving rise to a physiological role of brain <em>nesfatin</em>-<em>1</em> in the mediation of anxiety.

Keywords: Behavior; CRF; Depression; Gut-brain axis; NUCB2.

Neuropeptides are ancient endocrine components which have evolved to regulate many aspects of biology across the animal kingdom including behaviour, development and metabolism. To supplement current knowledge, we have utilized a transcriptome series describing larval development in the ornate spiny lobster, Panulirus ornatus. The biology of this animal has been leveraged to provide insights into the roles of molting, metamorphosis and metabolism across the neuropeptide family. We report an extensive list of neuropeptides across three distinct life phases of the animal. We show distinct groups of neuropeptides with differential expression between larval phases, indicating phase-specific roles for these peptides. For selected neuropeptides, we describe and discuss expression profiles throughout larval development and report predicted peptide cleavage sites and mature peptide sequences. We also report the neuropeptide <em>nesfatin</em> for the first time in a crustacean, and report secondary peptide products with a level of evolutionary conservation similar to the conventional mature peptide <em>nesfatin</em>-<em>1</em>, indicating a conserved role in these secondary products which are widely regarded as biologically inactive. In addition, we report a trend of downregulation in the neuropeptides as the animal undergoes extensive neural remodelling in fulfillment of metamorphosis. We suggest that this downregulation in neuropeptides relates to the brief, yet dramatic changes in morphology experienced by the central nervous system in the process of metamorphosis.

Preeclampsia (PE) is a leading cause of perinatal and maternal mortality. Considering that <em>Nesfatin</em>-<em>1</em> was reported to be downregulated in serum of PE patients, we aimed to explore the functional role of <em>Nesfatin</em>-<em>1</em> in trophoblast cells. Cell transfection was conducted to overexpress or inhibit <em>Nesfatin</em>-<em>1</em>, and its expression was measured by quantitative PCR. Cell proliferation, migration, and invasion abilities were determined employing CCK-8, flow cytometry, wound-healing, and transwell assays. Immunofluorescence assay was performed to detect E-cadherin and vimentin. ROS, MDA, and SOD levels were measured using their corresponding commercial kits. Western blot was used to identify the expression of vital kinases in PI3K/AKT/mTOR or GSK3β pathway and invasion-related proteins in trophoblast cells. <em>Nesfatin</em>-<em>1</em> knockdown significantly suppressed proliferation, migration, and invasion and increased cell arrest in G<em>1</em> phase, as well as downregulated expressions of MMP2/9 in HTR-8/SVneo cells. Besides, <em>Nesfatin</em>-<em>1</em> knockdown promoted the expression of E-cadherin and reduced the expression of vimentin. Additionally, the levels of ROS, MDA, and SOD were elevated upon <em>Nesfatin</em>-<em>1</em> knockdown. On the contrary, <em>Nesfatin</em>-<em>1</em> overexpression exerted the opposite effects. <em>Nesfatin</em>-<em>1</em> promoted the activation of PI3K/AKT/mTOR or GSK3β pathway, blocking of which reversed the promotive effects on trophoblast invasion and the inhibitory effects on oxidative stress of <em>Nesfatin</em>-<em>1</em> in HTR-8/SVneo cells. In short, this study revealed that <em>Nesfatin</em>-<em>1</em> promoted trophoblast cell proliferation, migration, invasion, and EMT and suppressed oxidative stress by activating PI3K/AKT/mTOR and AKT/GSK3β signaling pathway, laying the foundation for the development of therapeutic strategy for PE by targeting <em>Nesfatin</em>-<em>1</em>.

<strong class="sub-title"> Keywords: </strong> AKT; <em>Nesfatin</em>-<em>1</em>; Oxidative stress; Trophoblast invasion.

Various neuropeptides play an essential role in the nutrient sensing mechanism and related homeostasis. <em>Nesfatin</em>-<em>1</em> is a newly identified neuropeptide having anorectic activity, and <em>nesfatin</em>-<em>1</em>-containing neurons are widely distributed in the brain, including the hypothalamus and brain stem. Our previous study showed that dehydration-induced anorectic effects are mediated via the central <em>nesfatin</em>-<em>1</em> pathway in rats. Our recent studies have also shown that peripheral anorectic peptides (cholecystokinin-8, glucagon-like peptide-<em>1</em>, and leptin) and an antineoplastic agent (cisplatin) caused inhibition of feeding via the central <em>nesfatin</em>-<em>1</em> pathway in rats. <em>Nesfatin</em>-<em>1</em>-containing neurons in the central nervous system, in particular the hypothalamus and the brain stem, may mediate peripheral nutrient signals and regulate feeding behavior.

The aims of this study were to investigate the impacts of acute aerobic exercise on circulating levels of hormones associated with energy metabolism, namely leptin, <em>nesfatin</em>-<em>1</em> and irisin, in trained and untrained male subjects and to determine whether the timing of the exercise (i.e. morning or night) amplified these impacts. Thirty trained (<em>1</em>9.2±0.7 years) and 30 untrained (<em>1</em>9.5±0.6 years) male subjects performed two aerobic running exercises (3 days between tests) to 64-76% of the subjects' maximal heart rate for about 30 min. Pre- and post-exercise venous blood samples were taken and analysed for leptin, <em>nesfatin</em>-<em>1</em> and irisin using enzyme-linked immunosorbent assay (ELISA). Paired samples and independent samples t-tests were used to analyse data. Irisin levels increased in all the subjects (p<0.00<em>1</em>). In both groups, <em>nesfatin</em>-<em>1</em> levels increased significantly after the night-time exercise (p<0.05). Importantly, leptin and <em>nesfatin</em>-<em>1</em> levels varied among the trained and untrained groups: Both leptin and <em>nesfatin</em>-<em>1</em> levels increased in 4 (<em>1</em>3%) and <em>1</em>2 (40%) subjects, respectively, after the morning exercises, and they increased in 9 (30%) and <em>1</em>0 (33%) subjects, respectively, after the night-time exercise. They decreased in 5 (<em>1</em>6%) and 7 (23%) subjects, respectively, after the morning exercise and in 6 (20%) and 3 (<em>1</em>0%) subjects, respectively, after the night-time exercise. Exercise may result in increased energy consumption by altering irisin levels. However, due to variations among individuals, increasing leptin and <em>nesfatin</em>-<em>1</em> levels by reducing food intake may not be applicable.

<b>Background:</b> Major depressive disorder (MDD) is a mental health and emotional disorder that affects children and adolescents worldwide. This study aimed to evaluate serum <em>nesfatin</em>-<em>1</em>, ghrelin, and lipid levels as biological markers of adolescent MDD and their relationship with the severity of depression-anxiety and suicide risk in MDD. <b>Methods:</b>This study included 37 drug naïve adolescents between the ages of <em>1</em>2 and <em>1</em>8 who were diagnosed with a first episode MDD according to the Kiddie Schedule for Affective Disorders and Schizophrenia for School-Age Children-Present and Lifetime Version (K-SADS-PL) and DSM-V diagnostic criteria. Thirty-three healthy adolescents between the ages of <em>1</em>2 and <em>1</em>8 were included as the control group. The Children's Depression Inventory (CDI), Screen for Child Anxiety Related Disorders (SCARED), and Suicide Probability Scale (SPS) were used to evaluate the subjects in the MDD and control groups. In the first stage, serum <em>nesfatin</em>-<em>1</em>, ghrelin, and lipid levels were compared between the adolescents diagnosed with MDD and the control group. Next, the correlations between these levels and the CDI, SCARED, and SPS scores were evaluated. <b>Results:</b> <em>Nesfatin</em>-<em>1</em> levels were significantly lower in the MDD group than the control group (<i>p</i> < 0.00<em>1</em>) A positive correlation was found between the <em>nesfatin</em>-<em>1</em> levels and the SPS scores. <b>Conclusions:</b> This is the first study to evaluate <em>nesfatin</em>-<em>1</em> levels in adolescent depression, suggesting that <em>nesfatin</em>-<em>1</em>, ghrelin, total cholesterol, and low-density lipoprotein cholesterol (LDL) levels can be used as biomarkers in child-adolescent MDD. However, it is evident that further studies with larger samples and post-treatment measurements are needed.

<strong class="sub-title"> Keywords: </strong> Depression; adolescent; child; ghrelin; lipid; <em>nesfatin</em>-<em>1</em>.

Gestational diabetes mellitus (GDM) is a common disease in pregnant women, imposing risks on both mother and fetus. Dysregulated <em>nesfatin</em>-<em>1</em> has been observed in women with GDM, but the specific role of <em>nesfatin</em>-<em>1</em> underlying the pathological process of GDM is unclear. The main objective of this study is to investigate the role and the molecular mechanism of <em>nesfatin</em>-<em>1</em> in GDM. HTR-8/SVneo cells were treated with high glucose (HG)/high lipid (HL) to mimic the injured trophoblast of GDM in vitro. Cell viability, cytotoxicity and apoptosis were measured using CCK-8, LDH and TUNEL assays, respectively. The levels of inflammatory cytokines and antioxidant factors were detected using their commercial kits. ATP level and cytochrome c were determined with corresponding detecting kits. Quantitative real-time PCR and Western blot were performed to detect the expression of corresponding genes. The results showed that <em>nesfatin</em>-<em>1</em> was downregulated upon HG/HL stimulation. <em>Nesfatin</em>-<em>1</em> treatment greatly alleviated HG/HL-induced cell viability loss, cytotoxicity, inflammatory response, oxidative stress, and apoptosis in HTR-8/SVneo cells. In addition, <em>nesfatin</em>-<em>1</em> promoted ATP generation, reduced the leakage of cytochrome c from mitochondria to cytoplasm, and upregulated mitochondrial transcription factor A (TFAM) and nuclear respiratory factor <em>1</em> (NRF<em>1</em>), alleviating mitochondrial dysfunction. Furthermore, <em>nesfatin</em>-<em>1</em> inhibited p38 MAPK signaling. p79350, an agonist of p38 MAPK signaling, remarkably hindered the protective role of <em>nesfatin</em>-<em>1</em> in HG/HL-induced HTR-8/SVneo cells. In conclusion, <em>nesfatin</em>-<em>1</em> exerted a protective effect on GDM model in vitro, by regulating p38 MAPK signaling pathway, providing novel insights of treating GDM.

<strong class="sub-title"> Keywords: </strong> Gestational diabetes mellitus; mitochondria dysfunction; <em>nesfatin</em>-<em>1</em>; p38 MAPK.

<em>Nesfatin</em>-<em>1</em>, an 82-amino acid polypeptide derived from the precursor protein nucleobindin-2 (NUCB2), was first discovered in 2006 in the rat hypothalamus. The effects and distribution of <em>nesfatin</em>-<em>1</em> immunopositive neurons in the brain and spinal cord point towards a role of NUCB2/<em>nesfatin</em>-<em>1</em> in autonomic regulation. Therefore, studies which have been conducted to investigate the interplay between <em>nesfatin</em>-<em>1</em> and the autonomic nervous system were examined, and the outcomes of this research were summarized. NUCB2/<em>nesfatin</em>-<em>1</em> immunoreactivity is widely distributed in autonomic centers of the brain and spinal cord in both rodents and humans. In several regions of the hypothalamus, midbrain and brainstem, <em>nesfatin</em>-<em>1</em> modulates autonomic functions. On the other hand, the autonomic nervous system also influences the activity of <em>nesfatin</em>-<em>1</em> neurons. Here, the vagus nerve seems to be a crucial factor in the regulation of <em>nesfatin</em>-<em>1</em>. In summary, although data here is still sparse, there is a clear interplay between <em>nesfatin</em>-<em>1</em> and the autonomic nervous system, the precise clarification of which still requires further research to gain more insight into these complex relationships.

<strong class="sub-title"> Keywords: </strong> Food intake; Gut-brain axis; NUCB2; <em>Nesfatin</em>-<em>1</em>; Nucleobindin-2.

<em>Nesfatin</em>-<em>1</em> (encoded by NUCB2) is a cardiac peptide possessing protective activities against myocardial ischaemia/reperfusion (MI/R) injury. However, the regulation of NUCB2/<em>nesfatin</em>-<em>1</em> and the molecular mechanisms underlying its roles in MI/R injury are not clear. Here, by investigating a mouse MI/R injury model developed with transient myocardial ischaemia followed by reperfusion, we found that the levels of NUCB2 transcript and <em>nesfatin</em>-<em>1</em> amount in the heart were both decreased, suggesting a transcriptional repression of NUCB2/<em>nesfatin</em>-<em>1</em> in response to MI/R injury. Moreover, cardiac <em>nesfatin</em>-<em>1</em> restoration reduced infarct size, troponin T (cTnT) level and myocardial apoptosis, supporting its cardioprotection against MI/R injury in vivo. Mechanistically, the Akt/ERK pathway was activated, and in contrast, endoplasmic reticulum (ER) stress was attenuated by <em>nesfatin</em>-<em>1</em> following MI/R injury. In an in vitro system, similar results were obtained in <em>nesfatin</em>-<em>1</em>-treated H9c2 cardiomyocytes with hypoxia/reoxygenation (H/R) injury. More importantly, the treatment of wortmannin, an inhibitor of Akt/ERK pathway, abrogated <em>nesfatin</em>-<em>1</em> effects on attenuating ER stress and H/R injury in H9c2 cells. Furthermore, <em>nesfatin</em>-<em>1</em>-mediated protection against H/R injury also vanished in the presence of tunicamycin (TM), an ER stress inducer. Lastly, Akt/ERK inhibition reversed <em>nesfatin</em>-<em>1</em> effects on mouse ER stress and MI/R injury in vivo. Taken together, these findings demonstrate that NUCB2/<em>nesfatin</em>-<em>1</em> inhibits MI/R injury through attenuating ER stress, which relies on Akt/ERK pathway activation. Hence, our study provides a molecular basis for understanding how NUCB2/<em>nesfatin</em>-<em>1</em> reduces MI/R injury.

<strong class="sub-title"> Keywords: </strong> Akt/ERK pathway; NUCB2/<em>nesfatin</em>-<em>1</em>; endoplasmic reticulum stress; myocardial ischaemia/reperfusion injury.