Study of the long-lasting effects of ethanol consumption during adolescence on cannabinoid type 1 receptor-dependent synaptic transmission and plasticity in dentate gyrus synapses.

152 p.El consumo excesivo de alcohol, especialmente durante la adolescencia, es una preocupación generalizada de salud pública. Investigaciones en la última década han puesto de manifiesto que el consumo de etanol (EtOH) y el sistema endocannabinoide (eCB) interactúan recíprocamente para modificar la actividad y el comportamiento neuronal. Sin embargo, el impacto duradero de la ingesta de EtOH durante la adolescencia, en la localización y función del receptor Cannabinoide Tipo I (CB1) en el cerebro adulto y, en última instancia, en el comportamiento neurológico, sigue siendo desconocido. En este estudio de doctorado, se usaron ratones C57BL / 6J machos adultos para investigar la localización y la función del receptor CB1 en las sinapsis del trayecto perforante medial excitador (MPP) en la capa molecular dentada del hipocampo. Nos enfocamos en estas sinapsis porque: primero, integran el circuito tri-sináptico excitatorio del hipocampo involucrado en el aprendizaje y la memoria; segundo, las sinapsis MPP muestran una alta eficiencia en la activación neuronal; y tercero, pero no menos importante, la ingesta persistente de EtOH durante la adolescencia daña la corteza entorrinal y la circunvolución dentada y afecta la transmisión sináptica y la plasticidad. En particular, se estudió la implicación de losreceptores CB1 y el sistema eCB en la depresión a largo plazo de las sinapsis de células excitadoras de la MPP (CB1-eLTD), un tipo concreto de plasticidad sináptica. Luego, las consecuencias de la ingesta de EtOH adolescente en la transmisión y plasticidad sináptica de MPP se examinaron en la edad adulta después de la exposición de ratones C57BL / 6J machos adolescentes (día postnatal 32) a un consumo excesivo de alcohol de 4 días durante un período de 4 semanas (desde el día postnatal 32 hasta el 56) seguido de dos semanas de abstinencia de EtOH. Para alcanzar los objetivos propuestos, llevamos a cabo un enfoque experimental multidisciplinario basado en electrofisiología, inmunohistoquímica, comportamiento y técnicas de biología molecular. Los principales resultados de la Tesis Doctoral son: En primer lugar, la estimulación de baja frecuencia (10 min, 10 Hz) de la MPP desencadena CB1-eLTD en las sinapsis MPP. Este fenómeno de plasticidad es dependiente de los receptores de glutamato, en concreto de los receptores metabotrópicos del grupo I (mGluR), además requiere afluencia de calcio intracelular y síntesis de 2-araquidonoil-glicerol (2-AG). En segundo lugar, CB1-eLTD en las sinapsis MPP está ausente en ratones adultos después del consumo adolescente de EtOH. Además, la activación del receptor CB1 inhibe los potenciales postsinápticos excitatorios de campo (fEPSPs) evocados después de la estimulación de MPP en simulaciones adultas, pero no en ratones expuestos al EtOH. En tercer lugar, la ingesta de EtOH en adolescentes reduce significativamente la expresión del receptor CB1 en terminales sinápticos excitadores localizados en la zona de terminación MPP dentada, disminuye la unión basal [35S] guanosina-5 * -O- (3-tiotrifosfato) ([35S] GTP¿S) y la expresión de la subunidad G¿i2 y aumenta significativamente el ARNm y la proteína monoacilglicerol lipasa (MAGL) en el hipocampo adulto. En cuarto lugar, los ratones expuestos a EtOH muestran una memoria de reconocimiento, memoria espacial y memoria asociativa significativamente inferiores, así como una reducción significativa en la coordinación motora y el equilibrio después de dos semanas de la última sesión de EtOH. Sin embargo, no se detectan comportamientos de ansiedad o depresivos permanentes significativos. En quinto lugar, el aumento de 2-AG endógeno por el inhibidor de MAGL JZL184 rescata la CB1-eLTD y revierte la pérdida significativa de memoria de reconocimiento observada en ratones tratados con EtOH. En conclusión, el consumo excesivo de alcohol en adolescentes conduce a déficits en la transmisión excitatoria dependiente del receptor CB1 y la plasticidad en las sinapsis de células granulares de la vía perforante medial que se correlacionan con la pérdida de memoria y la alteración motora en ratones adultos. Además, tanto la CB1-eLTD como la memoria pueden recuperarse en ratones EtOH al aumentar los niveles endógenos de 2-A

Study of the long-lasting effects of ethanol consumption during adolescence on cannabinoid type 1 receptor-dependent synaptic transmission and plasticity in dentate gyrus synapses.

152 p.El consumo excesivo de alcohol, especialmente durante la adolescencia, es una preocupación generalizada de salud pública. Investigaciones en la última década han puesto de manifiesto que el consumo de etanol (EtOH) y el sistema endocannabinoide (eCB) interactúan recíprocamente para modificar la actividad y el comportamiento neuronal. Sin embargo, el impacto duradero de la ingesta de EtOH durante la adolescencia, en la localización y función del receptor Cannabinoide Tipo I (CB1) en el cerebro adulto y, en última instancia, en el comportamiento neurológico, sigue siendo desconocido. En este estudio de doctorado, se usaron ratones C57BL / 6J machos adultos para investigar la localización y la función del receptor CB1 en las sinapsis del trayecto perforante medial excitador (MPP) en la capa molecular dentada del hipocampo. Nos enfocamos en estas sinapsis porque: primero, integran el circuito tri-sináptico excitatorio del hipocampo involucrado en el aprendizaje y la memoria; segundo, las sinapsis MPP muestran una alta eficiencia en la activación neuronal; y tercero, pero no menos importante, la ingesta persistente de EtOH durante la adolescencia daña la corteza entorrinal y la circunvolución dentada y afecta la transmisión sináptica y la plasticidad. En particular, se estudió la implicación de losreceptores CB1 y el sistema eCB en la depresión a largo plazo de las sinapsis de células excitadoras de la MPP (CB1-eLTD), un tipo concreto de plasticidad sináptica. Luego, las consecuencias de la ingesta de EtOH adolescente en la transmisión y plasticidad sináptica de MPP se examinaron en la edad adulta después de la exposición de ratones C57BL / 6J machos adolescentes (día postnatal 32) a un consumo excesivo de alcohol de 4 días durante un período de 4 semanas (desde el día postnatal 32 hasta el 56) seguido de dos semanas de abstinencia de EtOH. Para alcanzar los objetivos propuestos, llevamos a cabo un enfoque experimental multidisciplinario basado en electrofisiología, inmunohistoquímica, comportamiento y técnicas de biología molecular. Los principales resultados de la Tesis Doctoral son: En primer lugar, la estimulación de baja frecuencia (10 min, 10 Hz) de la MPP desencadena CB1-eLTD en las sinapsis MPP. Este fenómeno de plasticidad es dependiente de los receptores de glutamato, en concreto de los receptores metabotrópicos del grupo I (mGluR), además requiere afluencia de calcio intracelular y síntesis de 2-araquidonoil-glicerol (2-AG). En segundo lugar, CB1-eLTD en las sinapsis MPP está ausente en ratones adultos después del consumo adolescente de EtOH. Además, la activación del receptor CB1 inhibe los potenciales postsinápticos excitatorios de campo (fEPSPs) evocados después de la estimulación de MPP en simulaciones adultas, pero no en ratones expuestos al EtOH. En tercer lugar, la ingesta de EtOH en adolescentes reduce significativamente la expresión del receptor CB1 en terminales sinápticos excitadores localizados en la zona de terminación MPP dentada, disminuye la unión basal [35S] guanosina-5 * -O- (3-tiotrifosfato) ([35S] GTP¿S) y la expresión de la subunidad G¿i2 y aumenta significativamente el ARNm y la proteína monoacilglicerol lipasa (MAGL) en el hipocampo adulto. En cuarto lugar, los ratones expuestos a EtOH muestran una memoria de reconocimiento, memoria espacial y memoria asociativa significativamente inferiores, así como una reducción significativa en la coordinación motora y el equilibrio después de dos semanas de la última sesión de EtOH. Sin embargo, no se detectan comportamientos de ansiedad o depresivos permanentes significativos. En quinto lugar, el aumento de 2-AG endógeno por el inhibidor de MAGL JZL184 rescata la CB1-eLTD y revierte la pérdida significativa de memoria de reconocimiento observada en ratones tratados con EtOH. En conclusión, el consumo excesivo de alcohol en adolescentes conduce a déficits en la transmisión excitatoria dependiente del receptor CB1 y la plasticidad en las sinapsis de células granulares de la vía perforante medial que se correlacionan con la pérdida de memoria y la alteración motora en ratones adultos. Además, tanto la CB1-eLTD como la memoria pueden recuperarse en ratones EtOH al aumentar los niveles endógenos de 2-A

Long-term effects of intermittent adolescent alcohol exposure in male and female rats

Alcohol is a serious public health concern that has a differential impact on individuals depending upon age and sex. Patterns of alcohol consumption have recently changed: heavy episodic drinking—known as binge-drinking—has become most popular among the youth. Herein, we aimed to investigate the consequences of intermittent adolescent alcohol consumption in male and female animals. Thus, Wistar rats were given free access to ethanol (20% in drinking water) or tap water for 2-h sessions during 3 days, and for an additional 4-h session on the 4th day; every week during adolescence, from postnatal day (pnd) 28–52. During this period, animals consumed a moderate amount of alcohol despite blood ethanol concentration (BEC) did not achieve binge-drinking levels. No withdrawal signs were observed: no changes were observed regarding anxiety-like responses in the elevated plus-maze or plasma corticosterone levels (pnd 53–54). In the novel object recognition (NOR) test (pnd 63), a significant deficit in recognition memory was observed in both male and female rats. Western Blot analyses resulted in an increase in the expression of synaptophysin in the frontal cortex (FC) of male and female animals, together with a decrease in the expression of the CB2R in the same brain region. In addition, adolescent alcohol induced, exclusively among females, a decrease in several markers of dopaminergic and serotonergic neurotransmission, in which epigenetic mechanisms, i.e., histone acetylation, might be involved. Taken together, further research is still needed to specifically correlate sex-specific brain and behavioral consequences of adolescent alcohol exposure

Environmental Enrichment Rescues Endocannabinoid-Dependent Synaptic Plasticity Lost in Young Adult Male Mice after Ethanol Exposure during Adolescence

Binge drinking (BD) is a serious health concern in adolescents as high ethanol (EtOH) consumption can have cognitive sequelae later in life. Remarkably, an enriched environment (EE) in adulthood significantly recovers memory in mice after adolescent BD, and the endocannabinoid, 2-arachydonoyl-glycerol (2-AG), rescues synaptic plasticity and memory impaired in adult rodents upon adolescent EtOH intake. However, the mechanisms by which EE improves memory are unknown. We investigated this in adolescent male C57BL/6J mice exposed to a drinking in the dark (DID) procedure four days per week for a duration of 4 weeks. After DID, the mice were nurtured under an EE for 2 weeks and were subjected to the Barnes Maze Test performed the last 5 days of withdrawal. The EE rescued memory and restored the EtOH-disrupted endocannabinoid (eCB)-dependent excitatory long-term depression at the dentate medial perforant path synapses (MPP-LTD). This recovery was dependent on both the cannabinoid CB1 receptor and group I metabotropic glutamate receptors (mGluRs) and required 2-AG. Also, the EE had a positive effect on mice exposed to water through the transient receptor potential vanilloid 1 (TRPV1) and anandamide (AEA)-dependent MPP long-term potentiation (MPP-LTP). Taken together, EE positively impacts different forms of excitatory synaptic plasticity in water- and EtOH-exposed brains.This research was funded by ISCIII (“RD16/0017/0012” to P.G.), co-funded by ERDF/ESF, “Investing in your future”; The Basque Government (IT1230-19 to P.G.); Ministry of Science and Innovation (PID2019-107548RB-I00 to P.G.); Ph.D. contract from MINECO (BES-2013-065057 to S.P.); Ph.D. contract from UPV/EHU (PIF 18/315 to L.L.), and Ph.D. contract from UPV/EHU (PIF 19/164 to M.S.)

Principales cambios en las Recomendaciones para la Resucitación del Consejo Europeo de Resucitación (ERC) 2015.

RESUMEN: Recientemente se han publicado las nuevas guías referentes a la Reanimación Cardiopulmonar (RCP) por parte del European Resuscitation Council (ERC). El mismo día, el Consejo Español de Resucitación Cardiopulmonar (CERCP) emitía una declaración oficial para manifestar su adhesión a estas recomendaciones. Esta misma declaración menciona “la necesidad de la aplicación ajustada de estas recomendaciones europeas, tanto en el ámbito asistencial como en el de la formación”. Aunque se mantienen la mayoría de las recomendaciones publicadas en el año 2010, es preciso conocer estos cambios para ajustarse a los nuevos conocimientos científicos. Para ello, este documento pretende resumir los principales cambios respecto a las recomendaciones anteriores con el objetivo de proporcionar una primera aproximación a los profesionales sanitarios.ABSTRACT: Recently, the new guidelines about cardiopulmonary resuscitation were published by the European Resuscitation Council (ERC). The same day, the Consejo Español de Resucitación Cardiopulmonar (CERCP) gave an announcement to declare the adhesion to these guidelines. This announcement brings up “the requirement to apply these European suggestions, both in the care services environment and in the formation environment”. Although most of the suggestions have had no changes with those suggestions published in 2010, the knowledge of these changes is needed to accommodate to the new knowledge. For this reason, this document pretends summarise the main changes in relation to the previous guidelines with the purpose to provide a first approach for healthcare professionals

The evolution of the ventilatory ratio is a prognostic factor in mechanically ventilated COVID-19 ARDS patients

Background: Mortality due to COVID-19 is high, especially in patients requiring mechanical ventilation. The purpose of the study is to investigate associations between mortality and variables measured during the first three days of mechanical ventilation in patients with COVID-19 intubated at ICU admission. Methods: Multicenter, observational, cohort study includes consecutive patients with COVID-19 admitted to 44 Spanish ICUs between February 25 and July 31, 2020, who required intubation at ICU admission and mechanical ventilation for more than three days. We collected demographic and clinical data prior to admission; information about clinical evolution at days 1 and 3 of mechanical ventilation; and outcomes. Results: Of the 2,095 patients with COVID-19 admitted to the ICU, 1,118 (53.3%) were intubated at day 1 and remained under mechanical ventilation at day three. From days 1 to 3, PaO2/FiO2 increased from 115.6 [80.0-171.2] to 180.0 [135.4-227.9] mmHg and the ventilatory ratio from 1.73 [1.33-2.25] to 1.96 [1.61-2.40]. In-hospital mortality was 38.7%. A higher increase between ICU admission and day 3 in the ventilatory ratio (OR 1.04 [CI 1.01-1.07], p = 0.030) and creatinine levels (OR 1.05 [CI 1.01-1.09], p = 0.005) and a lower increase in platelet counts (OR 0.96 [CI 0.93-1.00], p = 0.037) were independently associated with a higher risk of death. No association between mortality and the PaO2/FiO2 variation was observed (OR 0.99 [CI 0.95 to 1.02], p = 0.47). Conclusions: Higher ventilatory ratio and its increase at day 3 is associated with mortality in patients with COVID-19 receiving mechanical ventilation at ICU admission. No association was found in the PaO2/FiO2 variation

Early Maternal Deprivation Enhances Voluntary Alcohol Intake Induced by Exposure to Stressful Events Later in Life

In the present study, we aimed to assess the impact of early life stress, in the form of early maternal deprivation (MD, 24 h on postnatal day, pnd, 9), on voluntary alcohol intake in adolescent male and female Wistar rats. During adolescence, from pnd 28 to pnd 50, voluntary ethanol intake (20%, v/v) was investigated using the two-bottle free choice paradigm. To better understand the relationship between stress and alcohol consumption, voluntary alcohol intake was also evaluated following additional stressful events later in life, that is, a week of alcohol cessation and a week of alcohol cessation combined with exposure to restraint stress. Female animals consumed more alcohol than males only after a second episode of alcohol cessation combined with restraint stress. MD did not affect baseline voluntary alcohol intake but increased voluntary alcohol intake after stress exposure, indicating that MD may render animals more vulnerable to the effects of stress on alcohol intake. During adolescence, when animals had free access to alcohol, MD animals showed lower body weight gain but a higher growth rate than control animals. Moreover, the higher growth rate was accompanied by a decrease in food intake, suggesting an altered metabolic regulation in MD animals that may interact with alcohol intake

Endocannabinoid long-term depression revealed at medial perforant path excitatory synapses in the dentate gyrus

The endocannabinoid system modulates synaptic plasticity in the hippocampus, but a link between long-term synaptic plasticity and the type 1 cannabinoid (CB1) receptor at medial perforant path (MPP) synapses remains elusive. Here, immuno-electron microscopy in adult mice showed that similar to 26% of the excitatory synaptic terminals in the middle 1/3 of the dentate molecular layer (DML) contained CB1 receptors, and field excitatory postsynaptic potentials evoked by MPP stimulation were inhibited by CB1 receptor activation. In addition, MPP stimulation at 10 Hz for 10 min triggered CB, receptor-dependent excitatory long-term depression (eCB-eLTD) at MPP synapses of wild-type mice but not on CB1-knockout mice. This eCB-eLTD was group I mGluR-dependent, required intracellular calcium influx and 2-arachydonoyl-glycerol (2-AG) synthesis but did not depend on N-methyl-d-aspartate (NMDA) receptors. Overall, these results point to a functional role for CB1 receptors with eCB-eLTD at DML MPP synapses and further involve these receptors in memory processing within the adult brain.We thank all members of P. Grandes laboratory for their helpful comments, suggestions, and discussions during the performance of this study. The authors thank Giovanni Marsicano (INSERM, U1215 Neurocentre Magendie, Endocannabinoids and Neuroadaptation, Bordeaux, France. University de Bordeaux, France), Beat Lutz (Institute of Physiological Chemistry and German Resilience Center, University Medical Center of the Johannes Gutenberg University Mainz, Germany) and Susana Mato (Achucarro Basque Center for Neuroscience, Science Park of the UPV/EHU, Leioa, Vizcaya, Spain) for providing the CB1 receptor knock-out mice. This work was supported by MINECO/FEDER, UE (grant number SAF2015-65034-R to PG); The Basque Government (grant number BCG IT764-13 to PG); Red de Trastornos Adictivos, Instituto de Salud Carlos III (ISC-III) and European Regional Development Funds-European Union (ERDF-EU; grant RD16/0017/0012 to PG); PhD contract from MINECO (BES-2013-065057 to SP); Vanier Canada Graduate Scholarship (NSERC to CJF)

Intermittent ethanol exposure during adolescence impairs cannabinoid type 1 receptor-dependent long-term depression and recognition memory in adult mice.

Binge drinking is a significant problem in adolescent populations, and because of the reciprocal interactions between ethanol (EtOH) consumption and the endocannabinoid (eCB) system, we sought to determine if adolescent EtOH intake altered the localization and function of the cannabinoid 1 (CB1) receptors in the adult brain. Adolescent mice were exposed to a 4-day-per week drinking in the dark (DID) procedure for a total of 4 weeks and then tested after a 2-week withdrawal period. Field excitatory postsynaptic potentials (fEPSPs), evoked by medial perforant path (MPP) stimulation in the dentate gyrus molecular layer (DGML), were significantly smaller. Furthermore, unlike control animals, CB1 receptor activation did not depress fEPSPs in the EtOH-exposed animals. We also examined a form of excitatory long-term depression that is dependent on CB1 receptors (eCB-eLTD) and found that it was completely lacking in the animals that consumed EtOH during adolescence. Histological analyses indicated that adolescent EtOH intake significantly reduced the CB1 receptor distribution and proportion of immunopositive excitatory synaptic terminals in the medial DGML. Furthermore, there was decreased binding of [35S]guanosine-5*-O-(3-thiotriphosphate) ([35S] GTPγS) and the guanine nucleotide-binding (G) protein Gαi2 subunit in the EtOH-exposed animals. Associated with this, there was a significant increase in monoacylglycerol lipase (MAGL) mRNA and protein in the hippocampus of EtOH-exposed animals. Conversely, deficits in eCB-eLTD and recognition memory could be rescued by inhibiting MAGL with JZL184. These findings indicate that repeated exposure to EtOH during adolescence leads to long-term deficits in CB1 receptor expression, eCB-eLTD, and reduced recognition memory, but that these functional deficits can be restored by treatments that increase endogenous 2-arachidonoylglycerol