Inhibition of dedifferentiation in primary mouse hepatocytes in vitro to generate a functional hepatic study model

L'obésité et ses pathologies associées telles que la résistance à l'insuline, le diabète de type 2 et la stéatose hépatique non-alcoolique deviennent des enjeux de santé publique. Le foie est un organe majeur dans le contrôle de l'homéostasie du glucose corporel. À l'heure actuelle, les modèles de lignées cellulaires d'hépatome sont les principaux modèles d'étude in vitro disponibles pour approfondir la physiologie hépatique. Malheureusement, l'expression et les fonctions des protéines dans ces cellules diffèrent de la réalité in vivo. Les hépatocytes primaires représentent une plateforme attractive dans l'étude des maladies métaboliques car ils conservent la plupart des fonctions hépatiques in vivo. Une limitation fondamentale de la culture des hépatocytes primaires de souris correspond au déclin métabolique après leur isolement. En quelques jours, les cellules primaires se dédifférencient en une lignée cellulaire inférieure entraînant une perte des fonctions spécifiques et une modification de l'expression des gènes. Avec ce projet, nous avons étudié les hépatocytes primaires de souris en culture en présence de petites molécules modulant des voies spécifiques liées à la transition épithéliale à mésenchymateuse, un processus conduisant à la dédifférenciation dans les hépatocytes primaires isolés de souris. Nous avons constaté que des petits inhibiteurs chimiques du remodelage du cytosquelette préservaient partiellement l'expression de certains marqueurs hépatiques et épithéliaux tels que l'albumine, la Zonula-1 et l'occludine. De plus, nos résultats ont révélé que les hépatocytes primaires de souris en culture perdaient la réponse à l'insuline mais maintenaient la réponse au glucagon sur la gluconéogenèse bien que les valeurs soient beaucoup plus faibles après 7 jours de culture par rapport au jour 1. L'ensemble de nos résultats suggèrent que la réduction mécanique de la tension via l'inhibition du remodelage du cytosquelette pourrait être une voie à suivre pour générer un modèle in vitro fonctionnel d'hépatocytes primaires de souris à long-terme dans l'étude des maladies métaboliques.Obesity and its related pathologies insulin resistance, type 2 Diabetes and non-alcoholic fatty liver disease are becoming one of the major health hazards of modern world. The liver is a major organ in the control of body glucose homeostasis. Nowadays, hepatoma cell lines are in vitro study models available to further study liver physiology. Unfortunately, protein expression and functions in these cells differ from the in vivo reality. Primary hepatocytes are an attractive platform in the study of metabolic diseases because they retain most in vivo hepatic functions. A fundamental limitation of the culture of primary mouse hepatocytes is the metabolic decline taking place after isolation in these cells. Within few days isolated primary cells dedifferentiate into an inferior cell lineage losing specific functions and changing gene expression. Here, we studied primary mouse hepatocytes in culture in the presence of small molecules that modulate specific pathways related to epithelial to mesenchymal transition, a process leading to dedifferentiation in isolated primary mouse hepatocytes. We found that small chemical inhibitors of cytoskeletal changes partially preserved the expression of some hepatic and epithelial markers such as albumin, Zonula-1 and occludin. Furthermore, our results revealed that primary mouse hepatocytes in culture lost the response to insulin but maintained the response to glucagon on gluconeogenesis although the values decreased after 7 days in culture compared to day 1. Taken together, our results suggest that reducing mechanical tension by inhibiting cytoskeletal remodeling could be a way to follow to generate a functional in vitro model of long-term primary mouse hepatocytes to study metabolic diseases

Reclaiming human machine nature

Extending and modifying his domain of life by artifact production is one of the main characteristics of humankind. From the first hominid, who used a wood stick or a stone for extending his upper limbs and augmenting his gesture strength, to current systems engineers who used technologies for augmenting human cognition, perception and action, extending human body capabilities remains a big issue. From more than fifty years cybernetics, computer and cognitive sciences have imposed only one reductionist model of human machine systems: cognitive systems. Inspired by philosophy, behaviorist psychology and the information treatment metaphor, the cognitive system paradigm requires a function view and a functional analysis in human systems design process. According that design approach, human have been reduced to his metaphysical and functional properties in a new dualism. Human body requirements have been left to physical ergonomics or "physiology". With multidisciplinary convergence, the issues of "human-machine" systems and "human artifacts" evolve. The loss of biological and social boundaries between human organisms and interactive and informational physical artifact questions the current engineering methods and ergonomic design of cognitive systems. New developpment of human machine systems for intensive care, human space activities or bio-engineering sytems requires grounding human systems design on a renewed epistemological framework for future human systems model and evidence based "bio-engineering". In that context, reclaiming human factors, augmented human and human machine nature is a necessityComment: Published in HCI International 2014, Heraklion : Greece (2014

Effectiveness of auriculotherapy as a therapeutic alternative in patients with neurotic disorders

Introduction: the use of Natural and Traditional Medicine techniques is widespread in Guantanamo, however, no studies have been found that assess their results in patients with neurotic disorders. Objective: to evaluate the effectiveness of auriculotherapy in patients with neurotic disorders treated at the Provincial Psychiatric Hospital "Luis Ramirez Lopez" in Guantanamo in 2018. Method: a case-control study was conducted. The universe was made up of 323 patients, and a random sample (n=249) was selected. Variables were studied: sex, age, neurotic disorder, day of treatment in which it shows improvement, treatment outcome. Results: the highest number of patients was female in the control group (n=120) and cases (n=50), most affected patients (n=78) were in the age group of 48-58 year. Generalized anxiety disorders (n=182; 73.1%) were the most prevalent. Patients experienced improvement between 8 and 15 days of treatment (n= 60; 72.3%). Most of the cases were cured (n=80; 32.1%). Conclusions: the auriculotherapy as contributory treatment to the drug treatment is an effective tool to fight the neurotic disorders, it makes possible the remission of the symptoms and the cure of the patient in the smaller possible time

Efectividad de la auriculoterapia como alternativa terapéutica en pacientes con trastornos neuróticos

Introduction: the use of Natural and Traditional Medicine techniques is widespread in Guantanamo, however, no studies have been found that assess their results in patients with neurotic disorders. Objective: to evaluate the effectiveness of auriculotherapy in patients with neurotic disorders treated at the Provincial Psychiatric Hospital "Luis Ramirez Lopez" in Guantanamo in 2018. Method: a case-control study was conducted. The universe was made up of 323 patients, and a random sample (n=249) was selected. Variables were studied: sex, age, neurotic disorder, day of treatment in which it shows improvement, treatment outcome. Results: the highest number of patients was female in the control group (n=120) and cases (n=50), most affected patients (n=78) were in the age group of 48-58 year. Generalized anxiety disorders (n=182; 73.1%) were the most prevalent. Patients experienced improvement between 8 and 15 days of treatment (n= 60; 72.3%). Most of the cases were cured (n=80; 32.1%).Conclusions: the auriculotherapy as contributory treatment to the drug treatment is an effective tool to fight the neurotic disorders, it makes possible the remission of the symptoms and the cure of the patient in the smaller possible time.Introducción: en Guantánamo se generaliza la utilización de las técnicas de la Medicina Natural y Tradicional, sin embargo, no se encuentran estudios que valoren sus resultados en pacientes con trastornos neuróticos. Objetivo: evaluar la efectividad de la auriculoterapia en pacientes con trastornos neuróticos atendidos en el Hospital Psiquiátrico Provincial “Luis Ramírez López” en Guantánamo en el año 2018. Método: se realizó un estudio de casos y controles. El universo se conformó por 323 pacientes, y se seleccionó una muestra aleatoria (n=249). Se estudiaron las variables: sexo, edad, trastorno neurótico, día del tratamiento en que muestra mejoría, resultado del tratamiento. Resultados: el mayor número de pacientes fue del sexo femenino en el grupo de los controles (n=120) y los casos (n=50), los pacientes se encontraron más afectados en las edades comprendidas entre los 48 y 58 años (n=78). Los trastornos de ansiedad generalizada (n=182; 73,1 %) fueron los que más predominaron. Los casos experimentaron mejoría entre los 8 y 15 días (n= 60; 72,3 %). La mayor cantidad de los casos resultó curado (n=80; 32,1 %). Conclusiones: la auriculoterapia como tratamiento coadyuvante al tratamiento medicamentoso es una herramienta efectiva para combatir los trastornos neuróticos, posibilita la remisión de los síntomas y la curación del paciente en el menor tiempo posible

GABAB receptor-mediated activation of astrocytes by gamma-hydroxybutyric acid

The gamma-aminobutyric acid (GABA) metabolite gamma-hydroxybutyric acid (GHB) shows a variety of behavioural effects when administered to animals and humans, including reward/addiction properties and absence seizures. At the cellular level, these actions of GHB are mediated by activation of neuronal GABAB receptors (GABABRs) where it acts as a weak agonist. Because astrocytes respond to endogenous and exogenously applied GABA by activation of both GABAA and GABABRs, here we investigated the action of GHB on astrocytes on the ventral tegmental area (VTA) and the ventrobasal (VB) thalamic nucleus, two brain areas involved in the reward and proepileptic action of GHB, respectively, and compared it with that of the potent GABABR agonist baclofen. We found that GHB and baclofen elicited dose-dependent (ED50: 1.6 mM and 1.3 µM, respectively) transient increases in intracellular Ca2+ in VTA and VB astrocytes of young mice and rats, which were accounted for by activation of their GABABRs and mediated by Ca2+ release from intracellular store release. In contrast, prolonged GHB and baclofen exposure caused a reduction in spontaneous astrocyte activity and glutamate release from VTA astrocytes. These findings have key (patho)physiological implications for our understanding of the addictive and proepileptic actions of GHB

Gamma-hydroxybutyrate does not maintain self-administration but induces conditioned place preference when injected in the ventral tegmental area

Gamma-hydroxybutyric acid (GHB) is an endogenous brain substance that has diverse neuropharmacological actions, including rewarding properties in different animal species and in humans. As other drugs of abuse, GHB affects the firing of ventral tegmental neurons (VTA) in anaesthetized animals and hyperpolarizes dopaminergic neurons in VTA slices. However, no direct behavioural data on the effects of GHB applied in the VTA or in the target regions of its dopaminergic neurons, e.g. the nucleus accumbens (NAc), are available. Here, we investigated the effects of various doses of intravenous GHB in maintaining self-administration (from 0.001 to 10 mg/kg per infusion), and its ability to induce conditioned place preference (CPP) in rats when given orally (175-350 mg/kg) or injected directly either in the VTA or NAc (from 10 to 300 microg/0.5 microl per side). Our results indicate that while only 0.01 mg/kg per infusion GHB maintained self-administration, although not on every test day, 350 mg/kg GHB given orally induced CPP. CPP was also observed when GHB was injected in the VTA (30-100 microg/0.5 microl per side) but not in the NAc. Together with recent in-vitro findings, these results suggest that the rewarding properties of GHB mainly occur via disinhibition of VTA dopaminergic neurons

Gamma Hydroxybutyric Acid (GHB) for the Treatment of Alcohol Dependence: A Review

Gamma-hydroxybutyric acid (GHB) is a short-chain fatty acid structurally similar to the inhibitory neurotransmitter γ-aminobutyric acid. Clinical trials have demonstrated that 50–100 mg/kg of GHB fractioned into three or six daily doses is able to suppress alcohol withdrawal symptoms and facilitates the maintenance of abstinence from alcohol. These studies have also shown that GHB craving episodes are a very limited phenomenon (about 10–15%). Thus, physicians with access should consider the clinical efficacy of GHB as a valid pharmacological tool for the treatment of alcohol addiction