Search

The present study shows the development of computational algorithms to represent aircraft instruments such as the attitude indicator or the turn-and-slip indicator; moreover, the algorithms represent a magnetic compass and other instruments that function according to other physical theories. These instruments work by using the gyroscope and magnetic principles and help the pilot in navigation. These are considered to be the basic instruments required to provide location-related and positional information about the actual aircraft attitude. The algorithms developed in this study are capable of working in concordance with other instruments and the physical conditions established. The programming language used was C++ and the algorithms were compiled in independent files and subroutines for computational efficiency, eliminating unnecessary code. The display options were successfully tested. Additionally, an analysis that evaluated the error and approached flight simulation as a function of step time (Δt) is also described.

This manuscript describes the computational process to calculate an airplane path and display it in a 2D and 3D coordinate system on a computer screen. The airplane movement is calculated as a function of its dynamic’s conditions according to physical and logical theory. Here, the flight is divided into maneuvers and the aircraft conditions are defined as boundary conditions. Then the aircraft position is calculated using nested loops, which execute the calculation procedure at every step time (Δt). The calculation of the aircraft displacement is obtained as a function of the aircraft speed and heading angles. The simulator was created using the C++ programming language, and each part of the algorithm was compiled independently to reduce the source code, allow easy modification, and improve the programming efficiency. Aerial navigation involves very complex phenomena to be considered for an appropriate representation; moreover, in this manuscript, the influence of the mathematical approach to properly represent the aircraft flight is described in detail. The flight simulator was successfully tested by simulating some basic theoretical flights with different maneuvers, which include stationary position, running along the way, take off, and some movements in the airspace. The maximum aircraft speed tested was 120 km/h, the maximum maneuver time was 12 min, and the space for simulation was assumed to be without obstacles. Here, the geometrical description of path and speed is analyzed according to the symmetric and asymmetric results. Finally, an analysis was conducted to evaluate the approach of the numerical methods used; after that, it was possible to confirm that precision increased as the step time was reduced. According to this analysis, no more than 500 steps are required for a good approach in the calculation of the aircraft displacement.

Lead recycling is very important for reducing environmental pollution risks and damages. Liquid lead is recovered from exhaust batteries inside stirred batch reactors; the process requires melting to be cleaned. Nevertheless, it is necessary to establish parameters for evaluating mixing to improve the efficiency of the industrial practices. Computational fluid dynamics (CFD) has become a powerful tool to analyze industrial processes for reducing operating costs, avoiding potential damages, and improving the equipment’s performance. Thus, the present work is focused on simulating the fluid hydrodynamics inside a lead-stirred reactor monitoring the distribution of an injected tracer in order to find the best injection point. Then, different injected points are placed on a control plane for evaluation; these are evaluated one by one by monitoring the tracer concentration at a group of points inside the batch. The analyzed reactor is a symmetrical, vertical batch reactor with two geometrical sections: one cylindrical body and a semi-spherical bottom. Here, one impeller with four flat blades in a shaft is used for lead stirring. The tracer concentration on the monitoring points is measured and averaged for evaluating the efficiency inside the tank reactor. Hydrodynamics theory and a comparison between the concentration profiles and distribution of tracer curves are used to demonstrate both methods’ similarities. Then, the invariability of the tracer concentration on the monitoring points is adopted as the main parameter to evaluate the mixing, and the best injection point is found as a function of the shortest mixing time. Additionally, the influence of the impeller rotation speed is analyzed as an additional control parameter to improve industrial practices.

This work focuses on an analysis of hydrodynamics to improve the efficiency in a batch reactor for lead recycling. The study is based on computational fluid dynamics (CFD) methods, which are used to solve Navier–Stokes and Fick’s equations (continuity and momentum equations for understanding hydrodynamics and concentration for understanding distribution). The reactor analyzed is a tank with a dual geometry with a cylindrical body and a hemisphere for the bottom. This reactor is symmetrical vertically, and a shaft with four blades is used as an impeller for providing motion to the resident fluid. The initial resident fluid is static, and a tracer is defined in a volume inside to measure mixing efficiency, as is conducted in laboratory and industrial practices. Then, an evaluation of the mixing is performed by studying the tracer concentration curves at different evolution times. In order to understand the fluid flow hydrodynamics behavior with the purpose of identifying zones with rich and poor tracer concentrations, the tracer’s concentration was measured at monitoring points placed all around in a defined control plane of the tank. Moreover, this study is repeated independently to evaluate different injection points to determine the best one. Finally, it is proved that the selection of an appropriate injection point can reduce working times for mixing, which is an economically attractive motivation to provide proposals for improving industrial practices.

Abstract The global demand for fine-flavour cocoa has increased worldwide during the last years. Fine-flavour cocoa offers exceptional quality and unique fruity and floral flavour attributes of high demand by the world's elite chocolatiers. Several studies have highlighted the relevance of cocoa fermentation to produce such attributes. Nevertheless, little is known regarding the microbial interactions and biochemistry that lead to the production of these attributes on farms of industrial relevance, where traditional fermentation methods have been pre-standardized and scaled up. In this study, we have used metagenomic approaches to dissect on-farm industrial fermentations of fine-flavour cocoa. Our results revealed the presence of a shared core of nine dominant microorganisms (i.e. Limosilactobacillus fermentum, Saccharomyces cerevisiae, Pestalotiopsis rhododendri, Acetobacter aceti group, Bacillus subtilis group, Weissella ghanensis group, Lactobacillus_uc, Malassezia restricta and Malassezia globosa) between two farms located at completely different agro-ecological zones. Moreover, a community metabolic model was reconstructed and proposed as a tool to further elucidate the interactions among microorganisms and flavour biochemistry. Our work is the first to reveal a core of microorganisms shared among industrial farms, which is an essential step to process engineering aimed to design starter cultures, reducing fermentation times, and controlling the expression of undesirable phenotypes.

La elección de carrera es un proceso en el que un estudiante reconoce los intereses, actitudes y habilidades relacionadas con alguna profesión porque la considera atractiva, teniendo como resultado una decisión (Castañeda y Solorio, 2014). El objetivo del presente trabajo es sistematizar la experiencia del programa de orientación vocacional del Centro de Bachillerato Tecnológico Agropecuario 197, para elegir la carrera técnica con base en las competencias profesionales, a través de documentar y explicar el proceso seguido en el ciclo escolar anterior a la realización de este estudio, con el fin de proponer un modelo que contribuya a disminuir el índice de reprobación y aumentar el índice de titulación en subsiguientes años. La metodología tiene una perspectiva mixta, de carácter exploratorio con una lógica descriptiva y con fines de sistematización (Hernández, Fernández y Baptista, 2016). La muestra se compuso de 493 individuos equivalente al 90% de la población objetivo. El test consta de 28 reactivos, 25 relacionadas a las competencias profesionales de las carreras técnicas ofertadas en el plantel y tres para elegir su primera, segunda y tercera opción. Para el levantamiento de información se utilizó la plataforma Aplicaciones de Google herramienta Formularios. Los resultados encontrados indican que la primera opción que eligieron es la de Técnico Agropecuario, 36% (179), la segunda opción fue la Técnico en Producción Industrial de Alimentos 34% (167), y la tercera opción fue para la de Técnico en Biotecnología 26% (127). Con base a estos resultados se propone un modelo de intervención de orientación vocacional.

Simulation of the grain growth process, as a function of steel heat transfer conditions, is helpful for predicting grain structures of continuous cast steel products. Many authors have developed models based on numerical methods to simulate grain growth during metal solidification. Nevertheless, the anisotropic nature of grain structures makes necessary the employment of new mathematical methods such as chaos theory, fractals, and probabilistic and stochastic theories of simulation. The problem is significant for steelmakers to avoid defects in products and to control the steel microstructure during the continuous casting process. This work discusses the influence of nodal solidification times and computer algorithms on the dynamic formation of the chill, columnar, and equiaxed zones including physical phenomena such as nucleation and grain growth. Moreover, the model incorporates pre-nucleation and pre-growth routines in the original algorithm. There is a description of the influence of the mathematical parameter criteria and probabilities over the grain morphology obtained after solidification. Finally, an analysis of these algorithms elucidates the differences between these structures and those obtained from models considering only the solidification.

The current automation of steelmaking processes is capable of complete control through programmed hardware. However, many metallurgical and operating factors, such as heat transfer control, require further studies under industrial conditions. In this context, computer simulation has become a powerful tool for reproducing the effects of industrial constraints on heat transfer. This work reports a computational model to simulate heat removal from billets’ strands in the continuous casting process. This model deals with the non-symmetric cooling conditions of a billet caster. These cooling conditions frequently occur due to plugged nozzles in the secondary cooling system (SCS). The model developed simulates the steel thermal behavior for casters with a non-symmetric distribution of the sprays in the SCS using different boundary conditions to show possible heat transfer variations. Finally, the results are compared with actual temperatures from different casters to demonstrate the predictive capacity of this algorithm’s approach.

Ghrelin is the only known circulating orexigenic hormone. It increases food intake by interacting with hypothalamic and brainstem circuits involved in energy balance, as well as reward-related brain areas. A heightened gut-brain ghrelin axis is an emerging feature of certain eating disorders such as anorexia nervosa and Prader-Willi syndrome. In common obesity, ghrelin levels are lowered, whereas post-meal ghrelin levels remain higher than in lean individuals. Agents that interfere with ghrelin signalling have therapeutic potential for eating disorders, including obesity. However, most of these drugs are only in the preclinical phase of development. Data obtained so far suggest that ghrelin agonists may have potential in the treatment of anorexia nervosa, while ghrelin antagonists seem promising for other eating disorders such as obesity and Prader-Willi syndrome. However, large clinical trials are needed to evaluate the efficacy and safety of these drugs. [ABSTRACT FROM AUTHOR]

Las metas del milenio plantearon reducir a 400 millones de personas la incidencia del hambre en 2015; sin embargo y pese al reconocimiento universal del derecho a la alimentación el informe 2013 de la FAO sobre el estado mundial de la Agricultura y la Alimentación revela que 868 millones de personas, que equivale al 12.5% de la población mundial, están subordinadas en cuanto al consumo de energía alimentaria y 2000 millones de personas sufren carencia de micronutrientes. En el caso de México, la pérdida de la soberanía alimentaria se expresa en el hecho de que actualmente se importa casi 11 millones de toneladas y en carne de res se importa el 40% del consumo aparente, pese a que se exporta casi 2 millones de becerros a EUA. En este contexto, la presente investigación se justifica por el hecho de que no existe hasta la fecha un reconocimiento sistemático de las diferentes estrategias que están siguiendo los actores que participan en las redes agroalimentarias. En este caso se presenta el análisis de la red de valor del trigo en Sonora, como una modalidad del estudio de las cadenas agroalimentarias en México, lo que permite un acercamiento y comprensión de la estructura y dinámica de este cultivo, bajo un entorno de economía de libre mercado. Se observa la importancia a nivel estatal y nacional del trigo, especialmente porque durante las últimas décadas la producción de trigos cristalinos se orienta a la exportación y al sector de la porcicultura como parte de alimentos balanceados.

Abstract The vast microbial diversity on the planet represents an invaluable source for identifying novel activities with potential industrial and therapeutic application. In this regard, metagenomics has emerged as a group of strategies that have significantly facilitated the analysis of DNA from multiple environments and has expanded the limits of known microbial diversity. However, the functional characterization of enzymes, metabolites, and products encoded by diverse microbial genomes is limited by the inefficient heterologous expression of foreign genes. We have implemented a pipeline that combines NGS and Sanger sequencing as a way to identify fosmids within metagenomic libraries. This strategy facilitated the identification of putative proteins, subcloning of targeted genes and preliminary characterization of selected proteins. Overall, the in silico approach followed by the experimental validation allowed us to efficiently recover the activity of previously hidden enzymes derived from agricultural soil samples. Therefore, the methodology workflow described herein can be applied to recover activities encoded by environmental DNA from multiple sources.

El presente estudio se enfoca en el desarrollo de una metodología de fácil aplicación para la determinación de la incertidumbre para laboratorios de en sayo que utilicen la técnica de absorción atómica, siguiendo los criterios y recomendaciones de las normas ISO (International Organization for Standarization). La metodología desarrollada se aplicó para la determinación de cobre, y se consideraron las cuatro principales fuentes de incertidumbre: las involucradas en la curva de calibración del equipo, las atribuidas al pesaje de la muestra, las debidas a dos medidas de volumen en la solución final analizada y las atribuidas a las operaciones de dilución. El análisis químico fue de 50.85 ±0.62 (%P) de Cu, y del análisis de los coeficientes de sensibilidad se establece que la principal fuente de incertidumbre la representan las mediciones de volumen, por lo que un laboratorio que desee mejorar la confiabilidad en sus mediciones deberá de mejorar este aspecto.

El presente trabajo ilustra la descripción de rutinas computacionales basadas en diferentes modelos para la simulación de estructuras de grano. Métodos de Monte Carlo y de generación de números aleatorios son utilizados en conjunto a modelos geométricos y de autómatas celulares para generar diferentes morfologías en los granos y desplegarlas gráficamente en las pantallas de las computadoras. Además la influencia de los factores matemáticos y de los procesos computacionales empleados sobre la estructura de grano obtenida es descrita en detalle. Aunque los modelos geométricos son los mas simples y crean estructuras de grano similares a la realidad, tienen limitaciones debidas a la sencillez de su programación; mientras que los modelos de autómatas celulares y acoplados pueden ser empleados para generar estructuras más complejas y tomando como base reglas de generación y evolución.

En este trabajo se obtuvieron diferentes estados superficiales (diversos productos de corrosión) del acero API- X52 expuesto a la salmuera sintética (NACE-ID182). Se usaron diferentes técnicas electroquímicas, para diferentes tiempos de inmersión, así como voltamperometría cíclica a diferentes potenciales de inversión (-0.6 ≤ E+λ ≤ -0.1). La caracterización estructural se realizó por espectroscopia de impedancia electroquímica (EIE) y de electrónica de barrido (MEB). Los análisis de los espectros de impedancia de los diferentes estados superficiales formados se llevaron a cabo utilizando un circuito equivalente, asumiendo las mismas etapas para todos los procesos. Se observó que los valores de las impedancias real e imaginaria aumentan con el incremento del potencial al cual se formó el producto de corrosión. Las micrografías obtenidas mediante el MEB mostraron que la superficie dañada y el espesor de la película de los productos de corrosión están relacionados con el potencial aplicado.

En este trabajo, se desarrolla un estudio por simulación matemática para un baño agitado por un sistema de aspas planas simples, el cual es utilizado en el proceso de separación de plomo líquido. En este proceso, se realiza una inyección de trazador en diversos puntos axiales del baño. Se analiza la hidrodinámica del baño, las curvas de edades y tiempos de residencia resultantes en cada punto de inyección y de monitoreo, con el fin de obtener el mejor de los puntos de inyección sin tener que variar la geometría original del baño. Para mejorar la eficiencia del mezclado se analizan los perfiles de velocidades y concentración obtenidas para identificar zonas muertas y de poca recirculación. Los puntos de monitoreo para la inyección de trazador se consideran suficientes para obtener una dinámica apropiada de la homogenización del baño para fines de análisis.

Este trabajo muestra el desarrollo y la implementación de algoritmos computacionales basados en teoría del caos y autómatas celulares para la simulación del crecimiento de granos en secciones cuadradas de acero producidas por colada continua. Se desarrollaron tres algoritmos para la simulación de las estructuras de grano en las zonas perimetral, columnar y equiaxial; incluyen rutinas para estimular la nucleación y crecimiento del grano. Estas fueron compiladas separadamente al algoritmo principal con el objeto de hacer más eficiente el sistema de simulación. Los algoritmos analizan todos los nodos a cada paso, durante la simulación de cálculo y una interfase de despliegue gráfico muestra en la pantalla el proceso de solidificación. La teoría del caos fue empleada integrando un proceso aleatorio de selección. El resultado de este análisis es una imagen digital de celdas que representan a los granos (autómata celular) y es desplegada en la pantalla en función a un código numérico asignado a las posiciones nodales.

En este trabajo se evalúa el proceso de recuperación de metales comercialmente valiosos de catalizadores gastados y la disminución de la toxicidad de sus residuos. Los catalizadores gastados recibieron un tratamiento para la recuperación de níquel y molibdeno como metales de interés, que consistió de una tostación con carbonato de sodio seguida de lixiviación neutra y finalmente de lixiviación ácida en H2SO4 con la cual se logra una recuperación sustancial de estos metales. Los productos resultantes fueron caracterizados mediante las técnicas de Fluorescencia de Rayos X (FRX), Difracción de Rayos X (DRX), Microscopia Electrónica de Barrido (MEB), y Espectroscopia de Absorción Atómica (AA) para determinar cuantitativa y cualitativamente la eficiencia de la recuperación. Se concluye que la caracterización fisicoquímica de los diversos productos involucrados en el proceso permite detectar las características de los mismos, y así proponer un esquema de trabajo adecuado para la recuperación del metal valioso de los catalizadores gastados.

En este trabajo se estudia la influencia de los altos contenidos de molibdeno en dos aleaciones base (una con contenido alto de silicio y bajo en carbono, serie A; y la segunda con alto contenido de carbono y bajo silicio, serie B) sobre la microestructura y el comportamiento mecánico en piezas coladas en fundición esferoidal en espesores normales. Se enfatiza la composición química y características micro-estructurales tales como tamaño, distribución de nódulos, la relación de las fases formadas y propiedades mecánicas en el material obtenido. Se comprobó la influencia del contenido de molibdeno sobre la formación de nódulos en ambas series, así como de su morfología que es la característica deseada.

Background: Multiple neuropeptides, sometimes with opposing functions, can be produced from one precursor gene. To study the roles of the different neuropeptides encoded by one large precursor we developed a method to overexpress minigenes and establish local secretion.Results: We fused the signal peptide from the Von Willebrand Factor (VWF) to a furin site followed by a processed form of the Agouti related protein (AgRP), AgRP(83-132) or alpha-melanocyte stimulating hormone. In vitro, these minigenes were secreted and biologically active. Additionally, the proteins of the minigenes were not transported into projections of primary neurons, thereby ensuring local release. In vivo administration of VWF-AgRP(83-132), using an adeno-associated viral vector as a delivery vehicle, into the paraventricular hypothalamus increased body weight and food intake of these rats compared to rats which received a control vector.Conclusions: This study demonstrated that removal of the N-terminal part of full length AgRP and addition of a VWF signal peptide is a successful strategy to deliver neuropeptide minigenes to the brain and establish local neuropeptide secretion. [ABSTRACT FROM AUTHOR]

Accumulating evidence points to dysregulations of the Nucleus Accumbens (NAc) in eating disorders (ED), however its precise contribution to ED symptomatic dimensions remains unclear. Using chemogenetic manipulations in male mice, we found that activity of dopamine D1 receptor-expressing neurons of the NAc core subregion facilitated effort for a food reward as well as voluntary exercise, but decreased food intake, while D2-expressing neurons have opposite effects. These effects are congruent with D2-neurons being more active than D1-neurons during feeding while it is the opposite during running. Chronic manipulations of each subpopulations had limited effects on energy balance. However, repeated activation of D1-neurons combined with inhibition of D2-neurons biased behavior toward activity-related energy expenditure, whilst the opposite manipulations favored energy intake. Strikingly, concomitant activation of D1-neurons and inhibition of D2-neurons precipitated weight loss in anorexia models. These results suggest that dysregulations of NAc dopaminoceptive neurons might be at the core of EDs., (© 2024. The Author(s).)

Non-Invasive Brain Stimulation (NIBS) techniques and in particular, repetitive Transcranial Magnetic Stimulation (rTMS), are developing beyond mere clinical application. Although originally purposed for the treatment of resistant neuropsychiatric disorders, NIBS is also contributing to a deeper understanding of psychiatric disorders. rTMS is also changing the model of the disorder itself, from "mental" to one of neural connectivity. TMS allows the assessment of brain circuit excitability and eventually, of plastic changes affecting these circuits. While a clinical translational approach is, at the present time, the most adequate to meet the dimensional-circuit base model of the disorder, it refines the standard categorical classification of psychiatric disorders. The discovery of the fundamental importance of the balance between neuroplasticity and inflammation is also now explored through neuro-modulation findings consistently with the evidence of anti-inflammatory actions of the magnetic pulses. rTMS may activate, inhibit, or otherwise interfere with the activity of neuronal cortical networks, depending on stimulus frequency and intensity of brain-induced electric field. Of particular interest, yet still unclear, is how the relatively unspecific nature of TMS stimulation may lead to specific neuronal reorganization, as well as a definition of the TMS-triggered reorganization of functional brain modules, raising attention on the importance of the active participation of the patient to the treatment.. Configuration and state of consciousness of the subject have made subjective experience under treatment regain importance in the neuro-scientific Psychiatry based on the requirement of United States National Institute of Health (NIH) and the substantial importance of the consciousness state in the efficacy of the TMS treatment. By focusing on the subjective experience, a renaissance of the phenomenology offers Psychiatry an opportunity to become proficient and to distinguish itself from other disciplines. For all these reasons, TMS should be included in the cluster of the sub-specialties as a new "Super-Specialty" and an appropriate training course has to be inaugurated. Psychiatrists are nowadays multi-specialists, moving from a specialty to another, vs super-specialist. The cultivation of a properly trained cohort of TMS psychiatrists will better meet the challenges of treatment-resistant psychiatric conditions (disorders of connectivity), through appropriate and ethical practice, meanwhile facilitating an informed development and integration of additional emerging neuro-modulation techniques. The aim of this consensus paper is to underline the interdisciplinary nature of NIBS, that also encompasses the subjective experience and to point out the necessity of a neuroscience-applied approach to NIBS in the context of the European College of Neuro-psychopharmacology (ECNP)., Competing Interests: Declaration of Competing Interest SP has received grant from NIH outside the current work. AM has no conflicts of interest. PFP has received grant fees from Lundbeck LTD and honoraria from Angelini, Menarini, Lundbeck, Boehringer Ingelheim outside the current work. AML has received consultant fees from: Boehringer Ingelheim, Elsevier, Brainsway, Lundbeck Int. Neuroscience Foundation, Lundbeck A/S, The Wolfson Foundation, Bloomfield Holding Ltd, Shanghai Research Center for Brain Science, Thieme Verlag, Sage Therapeutics, v Behring Röntgen Stiftung, Fondation FondaMental, Janssen-Cilag GmbH, MedinCell, Brain Mind Institute, Agence Nationale de la Recherche, CISSN (Catania Internat. Summer School of Neuroscience), Daimler und Benz Stiftung, American Association for the Advancement of Science, Servier International. Additionally AML has received speaker fees from: Italian Society of Biological Psychiatry, Merz-Stiftung, Forum Werkstatt Karlsruhe, Lundbeck SAS France, BAG Psychiatrie Oberbayern, Klinik für Psychiatrie und Psychotherapie Ingolstadt, med Update GmbH, Society of Biological Psychiatry, Siemens Healthineers, Biotest AG. EV has received grants and served as consultant, advisor or CME speaker for the following entities (unrelated to the present work): AB-Biotics, Abbott, Allergan, Angelini, Dainippon Sumitomo Pharma, Ferrer, Gedeon Richter, Janssen, Lundbeck, Otsuka, Sage, Sanofi-Aventis, and Takeda. CSM has no conflicts of interest. RAHA has no conflicts of interest. SD has no conflicts of interest. BD in the last three years, has received lecture honoraria from Angelini, Lundbeck, Janssen, Pfizer, Otsuka, Neuraxpharm, Arcapharma and Livanova. CA has been a consultant to or has received honoraria or grants from Acadia, Angelini, Boehringer, Gedeon Richter, Janssen Cilag, Lundbeck, Minerva, Otsuka, Roche, Sage, Servier, Shire, Schering Plough, Sumitomo Dainippon Pharma, Sunovion and Takeda; CA has received funding from the Spanish Ministry of Science and Innovation. Instituto de Salud Carlos III (SAM16PE07CP1, PI16/02012, PI19/024), co-financed by ERDF Funds from the European Commission, “A way of making Europe”, CIBERSAM. Madrid Regional Government (B2017/BMD-3740 AGES-CM-2), European Union Structural Funds. European Union Seventh Framework Program under grant agreements FP7-4-HEALTH-2009-2.2.1-2-241909 (Project EU-GEI), FP7- HEALTH-2013-2.2.1-2-603196 (Project PSYSCAN) and FP7- HEALTH-2013-2.2.1-2-602478 (Project METSY); and European Union H2020 Program under the Innovative Medicines Initiative 2 Joint Undertaking (grant agreement No 115916, Project PRISM, and grant agreement No 777394, Project AIMS-2-TRIALS), Fundación Familia Alonso and Fundación Alicia Koplowitz. JZ Received grants / research supports from Lundbeck, Servier, Brainsway & Pfizer, received honoraria or consultation fees from Servier, Pfizer, Abbott, Lilly, Actelion, AstraZeneca, SunPharma, Roche and Brainsway and participated in company sponsored Lundbeck, Roche, Lilly, Servier, Pfizer, Speaker's bureau: Abbott, SunPharma and Brainsway, Stock shareholder: Nil, Spouse/partner: Nil. LC has no conflicts of interest., (Copyright © 2021. Published by Elsevier B.V.)

Melanin-Concentrating Hormone (MCH) is one of the most relevant orexigenic factors specifically located in the lateral hypothalamic area (LHA), with its physiological relevance demonstrated in studies using several genetically manipulated mice models. However, the central mechanisms controlling MCH-induced hyperphagia remain largely uncharacterized. Here, we show that central injection of MCH in mice deficient for kappa opoid receptor (k-OR) failed to stimulate feeding. To determine the hypothalamic area responsible for this MCH/k-OR interaction, we performed virogenetic studies and found that downregulation of k-OR by adeno-associated viruses (shOprk1-AAV) in LHA, but not in other hypothalamic nuclei, was sufficient to block MCH-induced food intake. Next, we sought to investigate the molecular signaling pathway within the LHA that mediates acute central MCH stimulation of food intake. We found that MCH activates k-OR and that increased levels of phosphorylated extracellular signal regulated kinase (ERK) are associated with downregulation of phospho-S6 Ribosomal Protein. This effect was prevented when a pharmacological inhibitor of k-OR was co-administered with MCH. Finally, the specific activation of the direct upstream regulator of S6 (p70S6K) in the LHA attenuated MCH-stimulated food consumption. Our results reveal that lateral hypothalamic k-OR system modulates the orexigenic action of MCH via the p70S6K/S6 pathway., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Attentional impairments and exaggerated impulsivity are key features of psychiatric disorders, such as attention-deficit/hyperactivity disorder, schizophrenia, and addiction. These deficits in attentional performance and impulsive behaviors have been associated with aberrant dopamine (DA) signaling, but it remains unknown whether these deficits result from enhanced DA neuronal activity in the midbrain. Here, we took a novel approach by testing the impact of chemogenetically activating DA neurons in the ventral tegmental area (VTA) or substantia nigra pars compacta (SNc) on attention and impulsivity in the five-choice serial reaction time task (5-CSRTT) in rats. We found that activation of DA neurons in both the VTA and SNc impaired attention by increasing trial omissions. In addition, SNc DA neuron activation decreased attentional accuracy. Surprisingly, enhanced DA neuron activity did not affect impulsive action in this task. These results show that enhanced midbrain DA neuronal activity induces deficits in attentional performance, but not impulsivity. Furthermore, DA neurons in the VTA and SNc have different roles in regulating attention. These findings contribute to our understanding of the neural substrates underlying attention deficits and impulsivity, and provide valuable insights to improve treatment of these symptoms.

Background: Food cues are omnipresent and may trigger overconsumption. In the past 2 decades, the prevalence of childhood obesity has increased dramatically. Because children's brains are still developing, especially in areas important for inhibition, children may be more susceptible than adults to tempting food cues., Objective: We examined potential developmental differences in children's and adults' responses to food cues to determine how these responses relate to weight status., Design: We included 27 children aged 10-12 y and 32 adults aged 32-52 y. Functional magnetic resonance imaging data were acquired during a food-viewing task in which unhealthy and healthy food pictures were presented., Results: Children had a stronger activation in the left precentral gyrus than did adults in response to unhealthy compared with healthy foods. In children, unhealthy foods elicited stronger activation in the right inferior temporal and middle occipital gyri, left precentral gyrus, bilateral opercular part of the inferior frontal gyrus, left hippocampus, and left middle frontal gyrus. Adults had stronger activation in the bilateral middle occipital gyrus and the right calcarine sulcus for unhealthy compared with healthy foods. Children with a higher body mass index (BMI) had lower activation in the bilateral dorsolateral prefrontal cortex while viewing unhealthy compared with healthy foods. In adults there was no correlation between BMI and neural response to unhealthy compared with healthy foods., Conclusions: Unhealthy foods might elicit more attention both in children and in adults. Children had stronger activation while viewing unhealthy compared with healthy foods in areas involved in reward, motivation, and memory. Furthermore, children activated a motivation and reward area located in the motor cortex more strongly than did adults in response to unhealthy foods. Finally, children with a higher BMI had less activation in inhibitory areas in response to unhealthy foods, which may mean they are more susceptible to tempting food cues. This trial was registered at www.trialregister.nl as NTR4255., (© 2016 American Society for Nutrition.)

Hyperactivity is a core symptom in various psychiatric disorders, including attention-deficit/hyperactivity disorder, schizophrenia, bipolar disorders, and anorexia nervosa. Although hyperactivity has been linked to dopaminergic signalling, the causal relationship between midbrain dopamine neuronal activity and locomotor hyperactivity remains unknown. In this study, we test whether increased dopamine neuronal activity is sufficient to induce locomotor hyperactivity. To do so, we used designer receptors exclusively activated by designer drugs (DREADD) to chemogenetically enhance neuronal activity in two main midbrain dopamine neuron populations, i.e. the ventral tegmental area (VTA) and substantia nigra pars compacta (SN), in TH:Cre rats. We found that activation of VTA dopamine neurons induced a pronounced and long-lasting hyperactive phenotype, whilst SN dopamine neuron activation only modestly increased home cage locomotion. Furthermore, this hyperactive phenotype was replicated by selective activation of the neuronal pathway from VTA to the nucleus accumbens (NAC). These results show a clear functional difference between neuronal subpopulations in the VTA and SN with regards to inducing locomotor hyperactivity, and suggest that the dopaminergic pathway from VTA to NAC may be a promising target for the treatment of hyperactivity disorders., (Copyright © 2016 Elsevier B.V. and ECNP. All rights reserved.)

Unlabelled: The opioid system is widely known to modulate the brain reward system and thus affect the behavior of humans and other animals, including feeding. We hypothesized that the hypothalamic opioid system might also control energy metabolism in peripheral tissues. Mice lacking the kappa opioid receptor (κOR) and adenoviral vectors overexpressing or silencing κOR were stereotaxically delivered in the lateral hypothalamic area (LHA) of rats. Vagal denervation was performed to assess its effect on liver metabolism. Endoplasmic reticulum (ER) stress was inhibited by pharmacological (tauroursodeoxycholic acid) and genetic (overexpression of the chaperone glucose-regulated protein 78 kDa) approaches. The peripheral effects on lipid metabolism were assessed by histological techniques and western blot. We show that in the LHA κOR directly controls hepatic lipid metabolism through the parasympathetic nervous system, independent of changes in food intake and body weight. κOR colocalizes with melanin concentrating hormone receptor 1 (MCH-R1) in the LHA, and genetic disruption of κOR reduced melanin concentrating hormone-induced liver steatosis. The functional relevance of these findings was given by the fact that silencing of κOR in the LHA attenuated both methionine choline-deficient, diet-induced and choline-deficient, high-fat diet-induced ER stress, inflammation, steatohepatitis, and fibrosis, whereas overexpression of κOR in this area promoted liver steatosis. Overexpression of glucose-regulated protein 78 kDa in the liver abolished hypothalamic κOR-induced steatosis by reducing hepatic ER stress., Conclusions: This study reveals a novel hypothalamic-parasympathetic circuit modulating hepatic function through inflammation and ER stress independent of changes in food intake or body weight; these findings might have implications for the clinical use of opioid receptor antagonists. (Hepatology 2016;64:1086-1104)., (© 2016 The Authors. (Hepatology published by Wiley Periodicals, Inc., on behalf of the American Association for the Study of Liver Diseases.)

Women ill with anorexia nervosa (AN) have been shown to exhibit altered cognitive functioning, particularly poor set-shifting (SS). In this study, we investigated whether brain activation in frontal and parietal regions during visual stimulus processing correlates with SS ability. Women currently ill with AN (AN; N=14), recovered women (REC; N=14) and healthy controls (HC; N=15), viewed alternating blocks of food and non-food pictures during functional magnetic resonance imaging (fMRI). The Berg's Card Sorting Task was completed outside the scanner to measure SS. A priori regions of interest (ROIs) were defined in frontal and parietal regions. The activation during visual stimulus processing in several ROIs correlated positively with poor SS ability in REC, particularly in the left dorsal anterior cingulate cortex (dACC). The correlations with poor SS ability were opposite in AN patients, particularly in the right dACC. These findings underscore that addressing heightened levels of cognitive control associated with higher frontal activation could reduce cognitive inflexibility in recovered women. In AN, greater activation in frontal and parietal regions might be necessary to perform at normal levels during various tasks. Thus, weight restoration could be necessary for AN patients prior to addressing cognitive inflexibility., (Copyright © 2016 Elsevier Ireland Ltd. All rights reserved.)

Unlabelled: Experience with sexual behavior causes cross-sensitization of amphetamine reward, an effect dependent on a period of sexual reward abstinence. We previously showed that ΔFosB in the nucleus accumbens (NAc) is a key mediator of this cross-sensitization, potentially via dopamine receptor activation. However, the role of mesolimbic dopamine for sexual behavior or cross-sensitization between natural and drug reward is unknown. This was tested using inhibitory designer receptors exclusively activated by designer drugs in ventral tegmental area (VTA) dopamine cells. rAAV5/hSvn-DIO-hm4D-mCherry was injected into the VTA of TH::Cre adult male rats. Males received clozapine N-oxide (CNO) or vehicle injections before each of 5 consecutive days of mating or handling. Following an abstinence period of 7 d, males were tested for amphetamine conditioned place preference (CPP). Next, males were injected with CNO or vehicle before mating or handling for analysis of mating-induced cFos, sex experience-induced ΔFosB, and reduction of VTA dopamine soma size. Results showed that CNO did not affect mating behavior. Instead, CNO prevented sexual experience-induced cross-sensitization of amphetamine CPP, ΔFosB in the NAc and medial prefrontal cortex, and decreases in VTA dopamine soma size. Expression of hm4D-mCherry was specific to VTA dopamine cells and CNO blocked excitation and mating-induced cFos expression in VTA dopamine cells. These findings provide direct evidence that VTA dopamine activation is not required for initiation or performance of sexual behavior. Instead, VTA dopamine directly contributes to increased vulnerability for drug use following loss of natural reward by causing neuroplasticity in the mesolimbic pathway during the natural reward experience., Significance Statement: Drugs of abuse act on the neural pathways that mediate natural reward learning and memory. Exposure to natural reward behaviors can alter subsequent drug-related reward. Specifically, experience with sexual behavior, followed by a period of abstinence from sexual behavior, causes increased reward for amphetamine in male rats. This study demonstrates that activation of ventral tegmental area dopamine neurons during sexual experience regulates cross-sensitization of amphetamine reward. Finally, ventral tegmental area dopamine cell activation is essential for experience-induced neural adaptations in the nucleus accumbens, prefrontal cortex, and ventral tegmental area. These findings demonstrate a role of mesolimbic dopamine in the interaction between natural and drug rewards, and identify mesolimbic dopamine as a key mediator of changes in vulnerability for drug use after loss of natural reward., (Copyright © 2016 the authors 0270-6474/16/369949-13$15.00/0.)

The central melanocortin (MC) system mediates its effects on food intake via MC3 (MC3R) and MC4 receptors (MC4R). Although the role of MC4R in meal size determination, satiation, food preference, and motivation is well established, the involvement of MC3R in the modulation of food intake has been less explored. Here, we investigated the role of MC3R on the incentive motivation for food, which is a crucial component of feeding behavior. Dopaminergic neurons within the ventral tegmental area (VTA) have a crucial role in the motivation for food. We here report that MC3Rs are expressed on VTA dopaminergic neurons and that pro-opiomelanocortinergic (POMC) neurons in the arcuate nucleus of the hypothalamus (Arc) innervate these VTA dopaminergic neurons. Our findings show that intracerebroventricular or intra-VTA infusion of the selective MC3R agonist γMSH increases responding for sucrose under a progressive ratio schedule of reinforcement, but not free sucrose consumption in rats. Furthermore, ex vivo electrophysiological recordings show increased VTA dopaminergic neuronal activity upon γMSH application. Consistent with a dopamine-mediated effect of γMSH, the increased motivation for sucrose after intra-VTA infusion of γMSH was blocked by pretreatment with the dopamine receptor antagonist α-flupenthixol. Taken together, we demonstrate an Arc POMC projection onto VTA dopaminergic neurons that modulates motivation for palatable food via activation of MC3R signaling.

Introduction: Conflicting data exist on sensitivity changes of the melanocortin system during diet-induced obesity. We hypothesized that melanocortin sensitivity depends on diet composition, in particular on the fat content rather than the level of obesity. The aim of this study was to determine the influence of diet composition on feeding responses to a melanocortin receptor agonist, using free-choice diets that differ in food components., Methods: Male Wistar rats were subjected to a chow (CHOW) diet or a free-choice (fc) diet of either chow, saturated fat and liquid sugar (fcHFHS), chow and saturated fat (fcHF), or chow and liquid sugar (fcHS) for 4 weeks. Melanocortin sensitivity was tested by measuring food intake following administration of the melanocortin 3/4 receptor agonist melanotan II (MTII) or vehicle in the lateral ventricle. In a separate experiment, proopiomelanocortin (POMC) and agouti-related protein (AgRP) mRNA levels were determined in the arcuate nucleus with in situ hybridization in rats subjected to the free-choice diets for 4 weeks., Results: Rats on the fcHFHS diet for 4 weeks show increased caloric intake and body weight gain compared to rats on the CHOW, fcHS and fcHF diet. Caloric intake and body weight gain was comparable between rats on the fcHF, fcHS, and CHOW diet. After 4 weeks diet, POMC and AgRP mRNA levels were not different between diet groups. MTII inhibited caloric intake to a larger extent in rats on the fcHF diet compared to rats on the CHOW, fcHFHS or fcHS diet. Moreover, the fat component was the most inhibited by MTII, and the sugar component the least., Conclusion: Rats on the fcHF diet show stronger food intake inhibition to the melanocortin receptor agonist MTII than rats on the CHOW, fcHS, and fcHFHS diet, which is independent of caloric intake and body weight gain. Our data point toward an important role for diet composition, particularly the dietary fat content, and not obesity in the sensitivity of the melanocortin system.

Background/objectives: The rewarding value of palatable foods contributes to overconsumption, even in satiated subjects. Midbrain dopaminergic activity in response to reward-predicting environmental stimuli drives reward-seeking and motivated behavior for food rewards. This mesolimbic dopamine (DA) system is sensitive to changes in energy balance, yet it has thus far not been established whether reward signaling of DA neurons in vivo is under control of hormones that signal appetite and energy balance such as ghrelin and leptin., Subjects/methods: We trained rats (n=11) on an operant task in which they could earn two different food rewards. We then implanted recording electrodes in the ventral tegmental area (VTA), and recorded from DA neurons during behavior. Subsequently, we assessed the effects of mild food restriction and pretreatment with the adipose tissue-derived anorexigenic hormone leptin or the orexigenic hormone ghrelin on VTA DA reward signaling., Results: Animals showed an increase in performance following mild food restriction (P=0.002). Importantly, food-cue induced DA firing increased when animals were food restricted (P=0.02), but was significantly attenuated after leptin pretreatment (P=0.00). While ghrelin did affect baseline DA activity (P=0.025), it did not affect cue-induced firing (P⩾0.353)., Conclusions: Metabolic signals, such as leptin, affect food seeking, a process that is dependent on the formation of cue-reward outcomes and involves midbrain DA signaling. These data show that food restriction engages the encoding of food cues by VTA DA neurons at a millisecond level and leptin suppresses this activity. This suggests that leptin is a key in linking metabolic information to reward signaling.

Background: Even though the evidence supporting the presence of a heritable component in the aetiology of anorexia nervosa (AN) is strong, the underlying genetic mechanisms remain poorly understood. The recent publication of a genome-wide association study (GWAS) of AN (Boraska, Mol Psychiatry, 2014) was an important step in genetic research in AN., Objective: To briefly sum up strengths and weaknesses of candidate-gene and genome-wide approaches, to discuss the genome-wide association studies of AN and to make predictions about the genetic architecture of AN by comparing it to that of schizophrenia (since the diseases share some similarities and genetic research in schizophrenia is more advanced)., Method: Descriptive literature review., Results: Despite remarkable efforts, the gene-association studies in AN did not advance our knowledge as much as had been hoped, although some results still await replication., Discussion: Continuous effort of participants, clinicians and researchers remains necessary to ensure that genetic research in AN follows a similarly successful path as in schizophrenia. Identification of genetic susceptibility loci provides a basis for follow-up studies., (© 2015 Wiley Periodicals, Inc.)

Understanding regulation and action of endogenous peptides, especially neuropeptides, which serve as inter- and intracellular signal transmitters, is key in understanding a variety of functional processes, such as energy balance, memory, circadian rhythm, drug addiction, etc. Therefore, accurate and reproducible quantification of these bioactive endogenous compounds is highly relevant. The biosynthesis of endogenous peptides, involving multiple possible trimming and modification events, hinders the de novo prediction of the active peptide sequences, making MS-based measurements very valuable in determining the actual active compounds. Here, we report an extended selected reaction monitoring (SRM)-based strategy to reproducibly and quantitatively monitor the abundances of a set of 15 endogenously occurring peptides from Rattus norvegicus hypothalamus. We demonstrate that SRM can be extended toward reproducible detection and quantification of peptides, bearing characteristics very different from tryptic peptides. We show that long peptide sequences, producing precursors with up to five and MS2 fragment ions with up to three charges, can be targeted by SRM on a triple quadrupole instrument. Using this approach to quantify endogenous peptide levels in hypothalami of animals subjected to different diets revealed several significant changes, most notably the significant upregulation of VGF-derived signaling peptide AQEE-30 upon high caloric feeding.

Altered mesolimbic dopamine signaling has been widely implicated in addictive behavior. For the most part, this work has focused on dopamine within the striatum, but there is emerging evidence for a role of the auto-inhibitory, somatodendritic dopamine D2 receptor (D2R) in the ventral tegmental area (VTA) in addiction. Thus, decreased midbrain D2R expression has been implicated in addiction in humans. Moreover, knockout of the gene encoding the D2R receptor (Drd2) in dopamine neurons has been shown to enhance the locomotor response to cocaine in mice. Therefore, we here tested the hypothesis that decreasing D2R expression in the VTA of adult rats, using shRNA knockdown, promotes addiction-like behavior in rats responding for cocaine or palatable food. Rats with decreased VTA D2R expression showed markedly increased motivation for both sucrose and cocaine under a progressive ratio schedule of reinforcement, but the acquisition or maintenance of cocaine self-administration were not affected. They also displayed enhanced cocaine-induced locomotor activity, but no change in basal locomotion. This robust increase in incentive motivation was behaviorally specific, as we did not observe any differences in fixed ratio responding, extinction responding, reinstatement or conditioned suppression of cocaine, and sucrose seeking. We conclude that VTA D2R knockdown results in increased incentive motivation, but does not directly promote other aspects of addiction-like behavior.

The role of the melanocortin (MC) system in feeding behavior is well established. Food intake is potently suppressed by central infusion of the MC 3/4 receptor agonist α-melanocyte stimulating hormone (α-MSH), whereas the MC 3/4 receptor inverse-agonist Agouti Related Peptide (AGRP) has the opposite effect. MC receptors are widely expressed in both hypothalamic and extra-hypothalamic brain regions, including nuclei involved in food reward and motivation, such as the nucleus accumbens (NAc) and the ventral tegmental area. This suggests that MCs modulate motivational aspects of food intake. To test this hypothesis, rats were injected intracerebroventricularly with α-MSH or AGRP and their motivation for sucrose was tested under a progressive ratio schedule of reinforcement. Food motivated behavior was dose-dependently decreased by α-MSH. Conversely, AGRP increased responding for sucrose, an effect that was blocked by pretreatment with the dopamine receptor antagonist α-flupenthixol. In contrast to progressive ratio responding, free intake of sucrose remained unaltered upon α-MSH or AGRP infusion. In addition, we investigated whether the effects of α-MSH and AGRP on food motivation were mediated by the NAc shell. In situ hybridization of MC3 and MC4 receptor expression confirmed that the MC4 receptor was expressed throughout the NAc, and injection of α-MSH and AGRP into the NAc shell caused a decrease and an increase in motivation for sucrose, respectively. These data show that the motivation for palatable food is modulated by MC4 receptors in the NAc shell, and demonstrate cross-talk between the MC and dopamine system in the modulation of food motivation.

The consumption of Western diets, high in sugar and saturated fat, is a crucial contributor to the alarming incidence of obesity and its associated morbidities. These diets have been reported to induce an inflammatory response in the hypothalamus, which promotes the development of central leptin resistance and obesity. This inflammatory signalling involves dynamic changes in the expression and activity of several mediators of the innate immune system, including toll-like receptor 4, IκB kinase-β/nuclear factor-κB, c-Jun N-terminal kinase, suppressor of cytokine signalling 3 and pro-inflammatory cytokines, as well as the induction of endoplasmic reticulum stress and autophagy defect. Although the exact cellular mechanisms remain incompletely understood, recent evidence suggests that the inflammatory response is at least mediated by interactions between neurons and non-neuronal cells such as microglia and astrocytes. Current evidence of the contribution of each inflammatory mediator to leptin resistance and diet-induced obesity (DIO), including their reciprocal interactions and cell-type-specific effects, is reviewed and integrated in a conceptual model. Based upon this model and pharmacological intervention studies, several inflammatory mediators are proposed to be promising therapeutic targets for the treatment of DIO., (© 2015 World Obesity.)

Anorexia nervosa (AN) is a severe mental disorder characterized by food restriction and weight loss. This study aimed to test the model posed by Brooks et al. (2012a,b) that women suffering from chronic AN show decreased food-cue processing activity in brain regions associated with energy balance and food reward (bottom-up; BU) and increased activity in brain regions associated with cognitive control (top-down; TD) when compared with long-term recovered AN (REC) and healthy controls (HC). Three groups of women, 15 AN (mean illness duration 7.8 ± 4.1 years), 14 REC (mean duration of recovery 4.7 ± 2.7 years) and 15 HC viewed alternating blocks of food and non-food images preceded by a short instruction during functional magnetic resonance imaging (fMRI), after fasting overnight. Functional region of interests (fROIs) were defined in BU (e.g., striatum, hippocampus, amygdala, hypothalamus, and cerebellum), TD (e.g., medial and lateral prefrontal cortex, and anterior cingulate), the insula, and visual processing areas (VPA). Food-cue processing activation was extracted from all fROIs and compared between the groups. In addition, functional connectivity between the fROIs was examined by modular partitioning of the correlation matrix of all fROIs. We could not confirm the hypothesis that BU areas are activated to a lesser extent in AN upon visual processing of food images. Among the BU areas the caudate showed higher activation in both patient groups compared to HC. In accordance with Brooks et al.'s model, we did find evidence for increased TD control in AN and REC. The functional connectivity analysis yielded two clusters in HC and REC, but three clusters in AN. In HC, fROIs across BU, TD, and VPA areas clustered; in AN, one cluster span across BU, TD, and insula; one across BU, TD, and VPA areas; and one was confined to the VPA network. In REC, BU, TD, and VPA or VPA and insula clustered. In conclusion, despite weight recovery, neural processing of food cues is also altered in recovered AN patients.

Food cues are omnipresent and may enhance overconsumption. In the last two decades the prevalence of childhood obesity has increased dramatically all over the world, largely due to overconsumption. Understanding children's neural responses to food may help to develop better interventions for preventing or reducing overconsumption. We aimed to determine which brain regions are concurrently activated in children/adolescents in response to viewing food pictures, and how these relate to adult findings. Two activation likelihood estimation (ALE) meta-analyses were performed: one with studies in normal weight children/adolescents (aged 8-18, 8 studies, 137 foci) and one with studies in normal weight adults (aged 18-45, 16 studies, 178 foci). A contrast analysis was performed for children/adolescents vs. adults. In children/adolescents, the most concurrent clusters were in the left lateral orbitofrontal cortex (OFC), the bilateral fusiform gyrus, and the right superior parietal lobule. In adults, clusters in similar areas were found. Although the number of studies for a direct statistical comparison between the groups was relatively low, there were indications that children/adolescents may not activate areas important for cognitive control. Overall, the number of studies that contributed to the significant clusters was moderate (6-75%). In summary, the brain areas most consistently activated in children/adolescents by food viewing are part of the appetitive brain network and overlap with those found in adults. However, the age range of the children studied was rather broad. This study offers important recommendations for future research; studies making a direct comparison between adults and children in a sufficiently narrow age range would further elucidate how neural responses to food cues change during development., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Single nucleotide polymorphisms (SNPs) clustered in the first intron of the fat mass and obesity-associated (FTO) gene has been associated with obesity. FTO expression is ubiquitous, with particularly high levels in the hypothalamic area of the brain. To investigate the region-specific role of FTO, AAV technology was applied to knockdown FTO in the ventromedial hypothalamus (VMH). No effect of FTO knockdown was observed on bodyweight or parameters of energy balance. Animals were exposed twice to an overnight fast, followed by a high-fat high-sucrose (HFHS) diet for 1 week. FTO knockdown did not result in a different response to the diets. A region-specific role for FTO in the VMH in the regulation of energy balance could not be found., (© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.)

The anorexigenic hormone leptin plays an important role in the control of food intake and feeding-related behavior, for an important part through its action in the hypothalamus. The adipose-derived hormone modulates a complex network of several intercommunicating orexigenic and anorexigenic neuropeptides in the hypothalamus to reduce food intake and increase energy expenditure. In this review we present an updated overview of the functional role of leptin in respect to feeding and feeding-related behavior per distinct hypothalamic nuclei. In addition to the arcuate nucleus, which is a major leptin sensitive hub, leptin-responsive neurons in other hypothalamic nuclei, including the, dorsomedial-, ventromedial- and paraventricular nucleus and the lateral hypothalamic area, are direct targets of leptin. However, leptin also modulates hypothalamic neurons in an indirect manner, such as via the melanocortin system. The dissection of the complexity of leptin's action on the networks involved in energy balance is subject of recent and future studies. A full understanding of the role of hypothalamic leptin in the regulation of energy balance requires cell-specific manipulation using of conditional deletion and expression of leptin receptors. In addition, optogenetic and pharmacogenetic tools in combination with other pharmacological (such as the recent discovery of a leptin receptor antagonist) and neuronal tracing techniques to map the circuit, will be helpful to understand the role of leptin receptor expressing neurons. Better understanding of these circuits and the involvement of leptin could provide potential sites for therapeutic interventions in obesity and metabolic diseases characterized by dysregulation of energy balance., (Copyright © 2014 Elsevier B.V. All rights reserved.)

Anorexia nervosa (AN) is a complex and heritable eating disorder characterized by dangerously low body weight. Neither candidate gene studies nor an initial genome-wide association study (GWAS) have yielded significant and replicated results. We performed a GWAS in 2907 cases with AN from 14 countries (15 sites) and 14 860 ancestrally matched controls as part of the Genetic Consortium for AN (GCAN) and the Wellcome Trust Case Control Consortium 3 (WTCCC3). Individual association analyses were conducted in each stratum and meta-analyzed across all 15 discovery data sets. Seventy-six (72 independent) single nucleotide polymorphisms were taken forward for in silico (two data sets) or de novo (13 data sets) replication genotyping in 2677 independent AN cases and 8629 European ancestry controls along with 458 AN cases and 421 controls from Japan. The final global meta-analysis across discovery and replication data sets comprised 5551 AN cases and 21 080 controls. AN subtype analyses (1606 AN restricting; 1445 AN binge-purge) were performed. No findings reached genome-wide significance. Two intronic variants were suggestively associated: rs9839776 (P=3.01 × 10(-7)) in SOX2OT and rs17030795 (P=5.84 × 10(-6)) in PPP3CA. Two additional signals were specific to Europeans: rs1523921 (P=5.76 × 10(-)(6)) between CUL3 and FAM124B and rs1886797 (P=8.05 × 10(-)(6)) near SPATA13. Comparing discovery with replication results, 76% of the effects were in the same direction, an observation highly unlikely to be due to chance (P=4 × 10(-6)), strongly suggesting that true findings exist but our sample, the largest yet reported, was underpowered for their detection. The accrual of large genotyped AN case-control samples should be an immediate priority for the field.

Eating disorders, such as anorexia nervosa (AN), bulimia nervosa (BN) and binge eating disorders (BED), are described as abnormal eating habits that usually involve insufficient or excessive food intake. Animal models have been developed that provide insight into certain aspects of eating disorders. Several drugs have been found efficacious in these animal models and some of them have eventually proven useful in the treatment of eating disorders. This review will cover the role of monoaminergic neurotransmitters in eating disorders and their pharmacological manipulations in animal models and humans. Dopamine, 5-HT (serotonin) and noradrenaline in hypothalamic and striatal regions regulate food intake by affecting hunger and satiety and by affecting rewarding and motivational aspects of feeding. Reduced neurotransmission by dopamine, 5-HT and noradrenaline and compensatory changes, at least in dopamine D2 and 5-HT(2C/2A) receptors, have been related to the pathophysiology of AN in humans and animal models. Also, in disorders and animal models of BN and BED, monoaminergic neurotransmission is down-regulated but receptor level changes are different from those seen in AN. A hypofunctional dopamine system or overactive α2-adrenoceptors may contribute to an attenuated response to (palatable) food and result in hedonic binge eating. Evidence for the efficacy of monoaminergic treatments for AN is limited, while more support exists for the treatment of BN or BED with monoaminergic drugs., (© 2014 The British Pharmacological Society.)

Neural growth regulator 1 (Negr1) is among the first common variants that have been associated with the regulation of body mass index. Using AAV technology directed to manipulate Negr1 expression in vivo, we find that decreased expression of Negr1 in periventricular hypothalamic areas leads to increases in body weight, presumably via increased food intake. Moreover, we observed that both increased and decreased levels of Negr1 lead to reduced locomotor activity and body temperature. In sum, our results provide further support for a role of hypothalamic expressed Negr1 in the regulation of energy balance., (© 2014 The Authors. Physiological Reports published by Wiley Periodicals, Inc. on behalf of the American Physiological Society and The Physiological Society.)

Rats on different free-choice (fc) diets for 1 week of either chow, saturated fat and liquid sugar (fcHFHS), chow and saturated fat (fcHF), or chow and liquid sugar (fcHS) have differential levels of neuropeptide Y (NPY) mRNA in the arcuate nucleus. Because these differences were not explained by plasma leptin levels but did predict subsequent feeding behaviour, in the present study, we first examined whether leptin sensitivity could explain these differences. Second, we focused on the role of NPY on feeding behaviour, and measured NPY mRNA levels and sensitivity to NPY after 4 weeks on the different choice diets. To determine leptin sensitivity, we measured food intake after i.p. leptin or vehicle injections in male Wistar rats subjected to the fcHFHS, fcHS, fcHF or Chow diets for 7 days. Next, we measured levels of arcuate nucleus NPY mRNA with in situ hybridisation in rats subjected to the choice diets for 4 weeks. Finally, we studied NPY sensitivity in rats subjected to the fcHFHS, fcHS, fcHF or Chow diet for 4 weeks by measuring food intake after administration of NPY or vehicle in the lateral ventricle. Leptin decreased caloric intake in rats on Chow, fcHS and fcHF but not in rats on the fcHFHS diet. After 4 weeks, rats on the fcHFHS diet remained hyperphagic, whereas fcHS and fcHF rats decreased caloric intake to levels similar to rats on Chow. By contrast to 1 week, after 4 weeks, levels of NPY mRNA were not different between the diet groups. Lateral ventricle administration of NPY resulted in higher caloric intake in fcHFHS rats compared to rats on the other choice diets or rats on Chow. Our data show that consuming a combination of saturated fat and liquid sugar results in leptin resistance and increased NPY sensitivity that is associated with persistent hyperphagia., (© 2014 British Society for Neuroendocrinology.)

To promote the efficient and safe application of adeno-associated virus (AAV) vectors as a gene transfer tool in the central nervous system (CNS), transduction efficiency and clearance were studied for serotypes commonly used to transfect distinct areas of the brain. As AAV2 was shown to transduce only small volumes in several brain regions, this study compares the transduction efficiency of three AAV pseudotyped vectors, namely AAV2/1, AAV2/5 and AAV2/8, in the ventromedial nucleus of the hypothalamus (VMH). No difference was found between AAV2/1 and AAV2/5 in transduction efficiency. Both AAV2/1 and AAV2/5 achieved a higher transduction rate than AAV2/8. One hour after virus administration to the brain, no viral particles could be traced in blood, indicating that no or negligible numbers of virions crossed the blood-brain barrier. In order to investigate survival of AAV in blood, clearance was determined following systemic AAV administration. The half-life of AAV2/1, AAV2/2, AAV2/5 and AAV2/8 was calculated by determining virus clearance rates from blood after systemic injection. The half-life of AAV2/2 was 4.2 minutes, which was significantly lower than the half-lives of AAV2/1, AAV2/5 and AAV2/8. With a half-life of more than 11 hours, AAV2/8 particles remained detectable in blood significantly longer than AAV2/5. We conclude that application of AAV in the CNS is relatively safe as no AAV particles are detectable in blood after injection into the brain. With a half-life of 1.67 hours of AAV2/5, a systemic injection with 1×109 genomic copies of AAV would be fully cleared from blood after 2 days.

Objective: Neuropeptide Y (NPY), given centrally augments food intake and the motivation to work for palatable food. Here, the brain regions were identified through which NPY increases food intake and motivation., Methods: NPY was infused into three brain regions implicated in food intake and motivation: the lateral hypothalamus (LH), nucleus accumbens shell (NAc), and ventral tegmental area (VTA). Motivation for sucrose was assessed using a progressive-ratio schedule of reinforcement in which the effort to obtain successive rewards increased incrementally. To disentangle the effects of NPY on motivation for palatable food from food consumption, free-feeding experiments were performed in which animals had ad libitum access to sucrose pellets., Results: Infusion of NPY into either VTA or NAc increased the motivation to respond for sucrose, whereas infusion of NPY in either NAc or LH increased sucrose consumption. In addition, the effect of intra-VTA NPY on motivation for food was attenuated after pretreatment with the dopamine receptor antagonist alpha-flupenthixol., Conclusions: Specific limbic substrates through which NPY influences consumption of and motivation for palatable food were identified by these data. The motivational effects of NPY are exerted through the VTA, its consummatory effects through the LH, and the NAc is involved in both., (Copyright © 2014 The Obesity Society.)

Objectives: Rats subjected to a free-choice high-fat high-sugar (fcHFHS) diet persistently overeat, exhibit increased food-motivated behavior and become overtly obese. Conversely, several studies using a non-choice (nc) high-energy diet showed only an initial increase in food intake with unaltered or reduced food-motivated behavior. This raises the question of the importance of choice in the persistence of hyperphagia in rats on a fcHFHS diet., Subjects: Meal patterns, food intake and body weight gain were studied in male Wistar rats on free-choice diets with fat and/or sugar and in rats on nc diets with fat and sugar (custom made with ingredients similar to the fcHFHS diet)., Results: Rats on a ncHFHS diet initially overconsumed, but reduced intake thereafter, whereas rats on a fcHFHS diet remained hyperphagic. Because half of the sugar intake in the fcHFHS group occurred during the inactive period, we next determined whether sugar intake during the light phase was a necessary requirement for hyperphagia, by restricting access to liquid sugar to either the light or dark period with unlimited access to fat and chow. Results showed that hyperphagia occurred irrespective of the timing of sugar intake. Meal pattern analysis revealed consumption of larger but fewer meals in the ncHFHS group, as well as the fcHF group. Interestingly, meal number was increased in all rats drinking liquid sugar (whether on a fcHFHS or a fcHS diet), whereas a compensatory decrease in meal size was only observed in the fcHS group, but not the fcHFHS group., Conclusion: We hereby show the importance of choice in the observation of fcHFHS diet-induced hyperphagia, which results in increases in meal number due to sugar drinking without any compensatory decrease in meal size. We thus provide a novel dietary model in rats that mimics important features of human overconsumption that have been ignored in rodent models of obesity.

We here describe a technique to transiently activate specific neural pathways in vivo. It comprises the combined use of a CRE-recombinase expressing canine adenovirus-2 (CAV-2) and an adeno-associated virus (AAV-hSyn-DIO-hM3D(Gq)-mCherry) that contains the floxed inverted sequence of the designer receptor exclusively activated by designer drugs (DREADD) hM3D(Gq)-mCherry. CAV-2 retrogradely infects projection neurons, which allowed us to specifically express hM3D(Gq)-mCherry in neurons that project from the ventral tegmental area (VTA) to the nucleus accumbens (Acb), the majority of which were dopaminergic. Activation of hM3D(Gq)-mCherry by intraperitoneal (i.p.) injections of clozapine-N-oxide (CNO) leads to increases in neuronal activity, which enabled us to specifically activate VTA to Acb projection neurons. The VTA to Acb pathway is part of the mesolimbic dopamine system and has been implicated in behavioral activation and the exertion of effort. Injections of all doses of CNO led to increases in progressive ratio (PR) performance. The effect of the lowest dose of CNO was suppressed by administration of a DRD1-antagonist, suggesting that CNO-induced increases in PR-performance are at least in part mediated by DRD1-signaling. We hereby validate the combined use of CAV-2 and DREADD-technology to activate specific neural pathways and determine consequent changes in behaviorally relevant paradigms.