Ghrelin treatment induces rapid and delayed increments of food intake : A heuristic model to explain ghrelin's orexigenic effects

Ghrelin is a stomach-derived peptide hormone with salient roles in the regulation of energy balance and metabolism. Notably, ghrelin is recognized as the most powerful known circulating orexigenic hormone. Here, we systematically investigated the effects of ghrelin on energy homeostasis and found that ghrelin primarily induces a biphasic effect on food intake that has indirect consequences on energy expenditure and nutrient partitioning. We also found that ghrelin-induced biphasic effect on food intake requires the integrity of Agouti-related peptide/neuropeptide Y-producing neurons of the hypothalamic arcuate nucleus, which seem to display a long-lasting activation after a single systemic injection of ghrelin. Finally, we found that different autonomic, hormonal and metabolic satiation signals transiently counteract ghrelin-induced food intake. Based on our observations, we propose a heuristic model to describe how the orexigenic effect of ghrelin and the anorectic food intake-induced rebound sculpt a timely constrain feeding response to ghrelin.Instituto Multidisciplinario de Biología Celula

The Nutritional Induction of COUP-TFII Gene Expression in Ventromedial Hypothalamic Neurons Is Mediated by the Melanocortin Pathway

BACKGROUND: The nuclear receptor chicken ovalbumin upstream promoter transcription factor II (COUP-TFII) is an important coordinator of glucose homeostasis. We report, for the first time, a unique differential regulation of its expression by the nutritional status in the mouse hypothalamus compared to peripheral tissues. METHODOLOGY/PRINCIPAL FINDINGS: Using hyperinsulinemic-euglycemic clamps and insulinopenic mice, we show that insulin upregulates its expression in the hypothalamus. Immunofluorescence studies demonstrate that COUP-TFII gene expression is restricted to a subpopulation of ventromedial hypothalamic neurons expressing the melanocortin receptor. In GT1-7 hypothalamic cells, the MC4-R agonist MTII leads to a dose dependant increase of COUP-TFII gene expression secondarily to a local increase in cAMP concentrations. Transfection experiments, using a COUP-TFII promoter containing a functional cAMP responsive element, suggest a direct transcriptional activation by cAMP. Finally, we show that the fed state or intracerebroventricular injections of MTII in mice induce an increased hypothalamic COUP-TFII expression associated with a decreased hepatic and pancreatic COUP-TFII expression. CONCLUSIONS/SIGNIFICANCE: These observations strongly suggest that hypothalamic COUP-TFII gene expression could be a central integrator of insulin and melanocortin signaling pathway within the ventromedial hypothalamus. COUP-TFII could play a crucial role in brain integration of circulating signal of hunger and satiety involved in energy balance regulation

Hypothalamic AgRP-neurons control peripheral substrate utilization and nutrient partitioning

Obesity-related diseases such as diabetes and dyslipidemia result from metabolic alterations including the defective conversion, storage and utilization of nutrients, but the central mechanisms that regulate this process of nutrient partitioning remain elusive. As positive regulators of feeding behaviour, agouti-related protein (AgRP) producing neurons are indispensible for the hypothalamic integration of energy balance. Here, we demonstrate a role for AgRP-neurons in the control of nutrient partitioning. We report that ablation of AgRP-neurons leads to a change in autonomic output onto liver, muscle and pancreas affecting the relative balance between lipids and carbohydrates metabolism. As a consequence, mice lacking AgRP-neurons become obese and hyperinsulinemic on regular chow but display reduced body weight gain and paradoxical improvement in glucose tolerance on high-fat diet. These results provide a direct demonstration of a role for AgRP-neurons in the coordination of efferent organ activity and nutrient partitioning, providing a mechanistic link between obesity and obesity-related disorders

Extracorporeal Membrane Oxygenation for Severe Acute Respiratory Distress Syndrome associated with COVID-19: An Emulated Target Trial Analysis.

RATIONALE: Whether COVID patients may benefit from extracorporeal membrane oxygenation (ECMO) compared with conventional invasive mechanical ventilation (IMV) remains unknown. OBJECTIVES: To estimate the effect of ECMO on 90-Day mortality vs IMV only Methods: Among 4,244 critically ill adult patients with COVID-19 included in a multicenter cohort study, we emulated a target trial comparing the treatment strategies of initiating ECMO vs. no ECMO within 7 days of IMV in patients with severe acute respiratory distress syndrome (PaO2/FiO2 <80 or PaCO2 ≥60 mmHg). We controlled for confounding using a multivariable Cox model based on predefined variables. MAIN RESULTS: 1,235 patients met the full eligibility criteria for the emulated trial, among whom 164 patients initiated ECMO. The ECMO strategy had a higher survival probability at Day-7 from the onset of eligibility criteria (87% vs 83%, risk difference: 4%, 95% CI 0;9%) which decreased during follow-up (survival at Day-90: 63% vs 65%, risk difference: -2%, 95% CI -10;5%). However, ECMO was associated with higher survival when performed in high-volume ECMO centers or in regions where a specific ECMO network organization was set up to handle high demand, and when initiated within the first 4 days of MV and in profoundly hypoxemic patients. CONCLUSIONS: In an emulated trial based on a nationwide COVID-19 cohort, we found differential survival over time of an ECMO compared with a no-ECMO strategy. However, ECMO was consistently associated with better outcomes when performed in high-volume centers and in regions with ECMO capacities specifically organized to handle high demand. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Ultrastable Silver Nanoparticles for Rapid Serology Detection of Anti-SARS-CoV-2 Immunoglobulins G.

Dipstick assays using silver nanoparticles (AgNPs) stabilized by a thin calix[4]arene-based coating were developed and used for the detection of Anti-SARS-CoV-2 IgG in clinical samples. The calixarene-based coating enabled the covalent bioconjugation of the SARS-CoV-2 Spike Protein via the classical EDC/sulfo-NHS procedure. It further conferred remarkable stability to the resulting bioconjugated AgNPs, as no degradation was observed over several months. In comparison with lateral-flow immunoassays (LFIAs) based on classical gold nanoparticles, our AgNP-based system constitutes a clear step forward, as the limit of detection for Anti-SARS-CoV-2 IgG was reduced by 1 order of magnitude and similar signals were observed with 10 times fewer particles. In real clinical samples, the AgNP-based dipstick assays showed impressive results: 100% specificity was observed for negative samples, while a sensitivity of 73% was determined for positive samples. These values match the typical sensitivities obtained for reported LFIAs based on gold nanoparticles. These results (i) represent one of the first examples of the use of AgNP-based dipstick assays in the case of real clinical samples, (ii) demonstrate that ultrastable calixarene-coated AgNPs could advantageously replace AuNPs in LFIAs, and thus (iii) open new perspectives in the field of rapid diagnostic&nbsp;tests.</p

Prebiotics Supplementation Impact on the Reinforcing and Motivational Aspect of Feeding.

Energy homeostasis is tightly regulated by the central nervous system which responds to nervous and circulating inputs to adapt food intake and energy expenditure. However, the rewarding and motivational aspect of food is tightly dependent of dopamine (DA) release in mesocorticolimbic (MCL) system and could be operant in uncontrolled caloric intake and obesity. Accumulating evidence indicate that manipulating the microbiota-gut-brain axis through prebiotic supplementation can have beneficial impact of the host appetite and body weight. However, the consequences of manipulating the implication of the microbiota-gut-brain axis in the control motivational and hedonic/reinforcing aspects of food are still underexplored. In this study, we investigate whether and how dietary prebiotic fructo-oligosaccharides (FOS) could oppose, or revert, the change in hedonic and homeostatic control of feeding occurring after a 2-months exposure to high-fat high-sugar (HFHS) diet. The reinforcing and motivational components of food reward were assessed using a two-food choice paradigm and a food operant behavioral test in mice exposed to FOS either during or after HFHS exposure. We also performed mRNA expression analysis for key genes involved in limbic and hypothalamic control of feeding. We show in a preventive-like approach, FOS addition of HFHS diet had beneficial impact of hypothalamic neuropeptides, and decreased the operant performance for food but only after an overnight fast while it did not prevent the imbalance in mesolimbic markers for DA signaling induced by palatable diet exposure nor the spontaneous tropism for palatable food when given the choice. However, when FOS was added to control diet after chronic HFHS exposure, although it did not significantly alter body weight loss, it greatly decreased palatable food tropism and consumption and was associated with normalization of MCL markers for DA signaling. We conclude that the nature of the diet (regular chow or HFHS) as well as the timing at which prebiotic supplementation is introduced (preventive or curative) greatly influence the efficacy of the gut-microbiota-brain axis. This crosstalk selectively alters the hedonic or motivational drive to eat and triggers molecular changes in neural substrates involved in the homeostatic and non-homeostatic control of body weight

COUP-TFII protein expression in adult mouse ventromedial hypothalamus.

<p>(A) Identification of the hypothalamic structures by Kluver Barrera staining. Black square: Arcuate nucleus, red square: Ventromedial hypothalamic nucleus. (B–F) Immunostaining of brain sections from adult mice using mouse monoclonal antibodies directed against COUP-TFII (PP-H7147-00, dilution 1∶50). The secondary antibody is coupled with the horse radish peroxydase and revealed with diaminobenzidine leading to a brown precipitate. The sections shown are from the same experiment with the same exposure times. Magnification ×20. (C) Secondary antibody incubation control for COUP-TFII antibody specificity (D) Enlargement from the VMH region, magnification ×40. (E) Enlargement of the arcuate nucleus region (ARH), magnification ×40. (F) Enlargement from the LHA region, magnification ×40. These pictures are representative of 6 independent experiments. (G–J) Immunofluorescence staining of adult mice hypothalamic cryostat sections, magnification ×20. (G) DAPI staining of the ARH-VMH region (blue fluorescence) (H) Identification of COUP-TFII positive neurons in the VMH-ARH region of adult mice using the COUP-TFII monoclonal antibody (green fluorescence) (I) Identification of COUP-TFI positive neurons in the VMH-ARH region of the hypothalamus using a COUP-TFI monoclonal antibody (red fluorescence) (J) Identification of COUP-TFI positive neurons in the cortex region of the same cryostat section than (I) as a positive control for COUP-TFI antibody. Scale bars  = 100 µm.</p

COUP-TFII protein co-localization studies with SF1 in the VMH.

<p>(A–G) Immunofluorescence staining of adult mice hypothalamic cryostat sections. (A–E) The sections shown are from the same experiment with the same exposure time (A) Identification of SF1 positive cells in the VMH by immunofluorescence using a SF1 polyclonal antibody (red fluorescence) (B) Identification of the COUP-TFII positive neurons in the VMH of adult mice using a COUP-TFII monoclonal antibody (green fluorescence) (C) Secondary antibody alone control for SF1 antibody specificity (D) Secondary antibody alone for COUP-TFII antibody specificity (E) Superimposed SF1 and COUP-TFII immunostaining. Magnification ×40. (F–G) The sections shown are from the same experiment and the same exposure time. (F) COUP-TFII immunostaining (red fluorescence) (G) Superimposed MC4-R and COUP-TFII immunofluorescences. Magnification ×100. These pictures are representative of 4 independent experiments. Scale bars  = 10 µm.</p

Regulation of hypothalamic COUP-TFII mRNA levels in vivo by activation of the melanocortin pathway.

<p>(A-C) RT-qPCR analysis of mRNA levels from hypothalamus of <i>ad libitum</i> fed mice receiving a ICV injection of CSF (□), or 18 h fasting mice receiving a ICV injection of CSF (grey bar), or 18 h fasting mice with a ICV injection of MTII (▪). (A) BDNF mRNA levels (B) Neuro D mRNA levels (C) COUP-TFII mRNA levels. Data are expressed as mean ± SEM (n = 5) *P≤0.05 **P ≤0.01 when compared to control fed mice receiving a CSF injection. § P≤0.05 when compared to control fasting mice receiving a CSF injection. (D) Glucose and plasma insulin concentrations from the different mouse models.</p