Pro-opiomelanocortin co-localizes with corticotropin-releasing factor in axon terminals of the noradrenergic nucleus locus coeruleus

We previously demonstrated that the opioid peptide, enkephalin, and corticotropin-releasing factor (CRF) are occasionally co-localized in individual axon terminals but more frequently converge on common dendrites in the locus coeruleus (LC). To further examine potential opioid co-transmitters in CRF afferents, we investigated the distribution of proopiomelanocortin (POMC), the precursor that yields the potent bioactive peptide, ß-endorphin, with respect to CRF immunoreactivity using immunofluorescence and immunoelectron microscopic analyses of the LC. Coronal sections were collected through the dorsal pontine tegmentum of rat brain and processed for immunocytochemical detection of POMC and CRF or tyrosine hydroxylase (TH). POMC-immunoreactive processes exhibited a distinct distribution within the LC as compared to the enkephalin family of opioid peptides. Specifically, POMC fibers were enriched in the ventromedial aspect of the LC with fewer fibers present dorsolaterally. Immunofluorescence microscopy showed frequent co-existence of POMC and CRF in varicose processes that overlapped TH-containing somatodendritic processes in the LC. Ultrastructural analysis showed POMC immunoreactivity in unmyelinated axons and axon terminals. Axon terminals containing POMC were filled with numerous large dense core vesicles. In sections processed for POMC and TH, approximately 29% of POMC-containing axon terminals (n = 405) targeted dendrites that exhibited immunogold-silver labeling for TH. Whereas, sections processed for POMC and CRF showed that 27% of POMC-labeled axon terminals (n = 657) also exhibited CRF immunoreactivity. Taken together, these data indicate that a subset of CRF afferents targeting the LC contain POMC and may be positioned to dually impact LC activity

An evaluation of neuroplasticity and behavior after deep brain stimulation of the nucleus accumbens in an animal model of depression.

BACKGROUND: Recent interest has demonstrated the nucleus accumbens (NAcc) as a potential target for the treatment of depression with deep brain stimulation (DBS). OBJECTIVE: To demonstrate that DBS of the NAcc is an effective treatment modality for depression and that chemical and structural changes associated with these behavioral changes are markers of neuroplasticity. METHODS: A deep brain stimulator was placed in the NAcc of male Wistar-Kyoto rats. Groups were divided into sham (no stimulation), intermittent (3 h/d for 2 weeks), or continuous (constant stimulation for 2 weeks). Exploratory and anxietylike behaviors were evaluated with the open-field test before and after stimulation. Tissue samples of the prefrontal cortex (PFC) were processed with Western blot analysis of markers of noradrenergic activity that included the noradrenergic synthesizing enzyme tyrosine hydroxylase. Analysis of tissue levels for catecholamines was achieved with high-performance liquid chromatography. Morphological properties of cortical pyramidal neurons were assessed with Golgi-Cox staining. RESULTS: Subjects undergoing intermittent and continuous stimulation of the NAcc exhibited an increase in exploratory behavior and reduced anxietylike behaviors. Tyrosine hydroxylase expression levels were decreased in the PFC after intermittent and continuous DBS, and dopamine and norepinephrine levels were decreased after continuous stimulation. Golgi-Cox staining indicated that DBS increased the length of apical and basilar dendrites in pyramidal neurons of the PFC. CONCLUSION: Deep brain stimulation induces behavioral improvement in and neurochemical and morphological alterations of the PFC that demonstrate changes within the circuitry of the brain different from the target area of stimulation. This observed dendritic plasticity may underlie the therapeutic efficacy of this treatment

Interaction of the mu-opioid receptor with GPR177 (Wntless) inhibits Wnt secretion: potential implications for opioid dependence

BACKGROUND: Opioid agonist drugs produce analgesia. However, long-term exposure to opioid agonists may lead to opioid dependence. The analgesic and addictive properties of opioid agonist drugs are mediated primarily via the mu-opioid receptor (MOR). Opioid agonists appear to alter neuronal morphology in key brain regions implicated in the development of opioid dependence. However, the precise role of the MOR in the development of these neuronal alterations remains elusive. We hypothesize that identifying and characterizing novel MOR interacting proteins (MORIPs) may help to elucidate the underlying mechanisms involved in the development of opioid dependence. RESULTS: GPR177, the mammalian ortholog of Drosophila Wntless/Evi/Sprinter, was identified as a MORIP in a modified split ubiquitin yeast two-hybrid screen. GPR177 is an evolutionarily conserved protein that plays a critical role in mediating Wnt protein secretion from Wnt producing cells. The MOR/GPR177 interaction was validated in pulldown, coimmunoprecipitation, and colocalization studies using mammalian tissue culture cells. The interaction was also observed in rodent brain, where MOR and GPR177 were coexpressed in close spatial proximity within striatal neurons. At the cellular level, morphine treatment caused a shift in the distribution of GPR177 from cytosol to the cell surface, leading to enhanced MOR/GPR177 complex formation at the cell periphery and the inhibition of Wnt protein secretion. CONCLUSIONS: It is known that chronic morphine treatment decreases dendritic arborization and hippocampal neurogenesis, and Wnt proteins are essential for these processes. We therefore propose that the morphine-mediated MOR/GPR177 interaction may result in decreased Wnt secretion in the CNS, resulting in atrophy of dendritic arbors and decreased neurogenesis. Our results demonstrate a previously unrecognized role for GPR177 in regulating cellular response to opioid drugs

Pediatric liver transplantation: A single center experience spanning 20 years

Background. Survival after liver transplantation has improved significantly over the last decade with pediatric recipients faring better than adults. The 20-year experience of pediatric liver transplantation at Children's Hospital of Pittsburgh is reported in terms of patient survival; graft survival in relation to age, gender, and immunosuppressive protocols; causes of death; and indications for retransplantation. Method. From March 1981 to April 1998, 808 children received liver transplants at Children's Hospital of Pittsburgh. All patients were followed until March 2001, with a mean follow-up of 12.2±3.9 years (median= 12.6; range=2.9-20). There were 405 female (50.2%) and 403 male (49.8%) pediatric recipients. Mean age at transplant was 5.3±4.9 years (mean=3.3; range 0.04-17.95), with 285 children (25.3%) being less than 2 years of age at transplant. Cyclosporine (CsA)-based immunosuppression was used before November 1989 in 482 children (50.7%), and the subsequent 326 recipients (40.3%) were treated with tacrolimus-based immunosuppression. Actuarial survival was calculated using the Kaplan-Meier statistical method. Differences in survival were calculated by log-rank analysis. Results. Overall patient survival at 1, 5, 10, 15 and 20 years was 77.1%, 72.6%, 69.4%, 65.8% and 64.4%, respectively. There was no difference in survival for male or female patients at any time point. At up to 10 years posttransplant, the survival for children greater than 2 years of age (79.5%, 75.7%, and 71.6% at 1, 5, and 10 years, respectively) was slightly higher than those at less than 2 years of age (72.6%, 66.9%, and 65.3% at 1, 5, and 10 years, respectively). However, at 15 and 20 years posttransplant, survival rates were similar (>2 years=67.3% and 65.8%; <2 years=64.1% and 64.1%). A significant difference in survival was seen in CsA-based immunosuppression (71.2%, 68.1%, 65.4%, and 61%) versus tacrolimus-based immunosuppression (85.8%, 84.7%, 83.3%, and 82.9%) at 1, 3, 5, and 10 years, respectively (P=0.0001). The maximum difference in survival was noted in the first 3 months between CsA and tacrolimus; thus, indicating there may have been other factors (nonimmunological factors) involved in terms of donor and recipient selection and technical issues. The mean annual death rate beyond 2 years posttransplant was 0.47%, with the mean annual death rate for patients who received tacrolimus-based immunosuppression being significantly lower than those who received CsA-based immunosuppression (0.14% vs. 0.8%; P=0.001). The most common etiologies of graft loss were hepatic artery thrombosis (33.4%), acute or chronic rejection (26.6%), and primary nonfunction (16.7%). Of note, retransplantation for graft loss because of acute or chronic rejection occurred only in those patients who received CsA-based immunosuppression. Conclusion. The overall 20-year actuarial survival for pediatric liver transplantation is 64%. Survival has increased by 20% in the last 12 years with tacrolimus-based immunosuppression. although this improvement may be the result of several factors, retransplantation as a result of acute or chronic rejection has been completely eliminated in patient treated with tacrolimus

Single-Cell Glia and Neuron Gene Expression in the Central Amygdala in Opioid Withdrawal Suggests Inflammation With Correlated Gut Dysbiosis.

Drug-seeking in opioid dependence is due in part to the severe negative emotion associated with the withdrawal syndrome. It is well-established that negative emotional states emerge from activity in the amygdala. More recently, gut microflora have been shown to contribute substantially to such emotions. We measured gene expression in single glia and neurons gathered from the amygdala using laser capture microdissection and simultaneously measured gut microflora in morphine-dependent and withdrawn rats to investigate drivers of negative emotion in opioid withdrawal. We found that neuroinflammatory genes, notabl

Efectos de las estrategias empresariales de los AGENTES BANPRO en la SATISFACCION DE LOS CLIENTES en la ciudad de Estelí en el año 2014

El presente trabajo determina los efectos de las estrategias empresariales de los agentes BANPRO para diseñar  propuestas que mejore la satisfacción de los clientes y agentes en  la ciudad de Estelí en el año 2014. El desarrollo de este estudio se hizo tomando en cuenta el enfoque cualitativo. El universo de estudio fue 23 agentes los cuales se encuentran ubicados en el municipio, con un tipo de muestreo no probabilístico porque los sujetos de consulta son elegidos en base a criterios selectivos como: Propietarios de establecimientos bancarios, clientes de BANPRO que usan este servicio, disponibilidad de las personas en brindar la información. Los instrumentos utilizados en el estudio fue la entrevista, guía de observación y por carácter del segundo objetivo se aplico encuestas dirigidas a clientes. Una limitante para el crecimiento de estos agentes es la publicidad muchos clientes desconoce este servicio y es inevitable su desconfianza al utilizarlo por primera vez, otra es la ubicación del POS dentro del local porque no garantiza la seguridad del cliente, haciendo que los usuarios recurran a las sucursales bancarias para realizar sus pagos.Es fundament al que BANPRO diseñe planes estratégicos para mejorar las limitantes que tienen  como diseñar ventanillas para estos establecimientos y la importancia de invertir en publicidad para dar a conocer lo que se oferta.Palabras claves: Banco, BANPRO,  Estrategia Empresariales, Satisfacción de clientes, Estel

Efectos de las estrategias empresariales de los AGENTES BANPRO en la SATISFACCION DE LOS CLIENTES en la ciudad de Estelí en el año 2014

The present study determines the effects of corporate strategies of BANPRO Agents (Bank of Production) to design proposals to improve customer satisfaction and agents’ service in the city of Estelí in 2014. This study is qualitative. The study group was 23 agents located in the city of Estelí. The sampling was non-probability, considering the following criteria: owners of banking establishments, customers using this service, and availability of people to provide information. Interviews, surveys and observation guides were applied.This study shows that the lack of publicity is one of the main constraints for the growth of BANPRO agents since many customers are unaware of this service and it is inevitable their distrust when using it for the first time. Another limitation is the location within the buildings because it does not ensure customer safety.It is recommended that the bank designs strategic plans to improve the constraints they have such as to design windows for these establishments, and the importance of investing in advertisement to make notorious what it is offered.El presente trabajo determina los efectos de las estrategias empresariales de los agentes BANPRO para diseñar propuestas que mejore la satisfacción de los clientes y agentes en la ciudad de Estelí en el año 2014. El desarrollo de este estudio se hizo tomando en cuenta el enfoque cualitativo. El universo de estudio fue 23 agentes los cuales se encuentran ubicados en el municipio, con un tipo de muestreo no probabilístico porque los sujetos de consulta son elegidos en base a criterios selectivos como: Propietarios de establecimientos bancarios, clientes de BANPRO que usan este servicio, disponibilidad de las personas en brindar la información.Los instrumentos utilizados en el estudio fue la entrevista, guía de observación y por carácter del segundo objetivo se aplico encuestas dirigidas a clientes. Una limitante para el crecimiento de estos agentes es la publicidad muchos clientes desconoce este servicio y es inevitable su desconfianza al utilizarlo por primera vez, otra es la ubicación del POS dentro del local porque no garantiza la seguridad del cliente, haciendo que los usuarios recurran a las sucursales bancarias para realizar sus pagos.Es fundament al que BANPRO diseñe planes estratégicos para mejorar las limitantes que tienen como diseñar ventanillas para estos establecimientos y la importancia de invertir en publicida para dar a conocer lo que se oferta

Longitudinal Mediators of Relations Between Family Violence and Adolescent Dating Aggression Perpetration: Family Violence and Adolescent Dating Aggression

Few longitudinal studies have examined the pathways through which family violence leads to dating aggression. In the current study the authors used 3 waves of data obtained from 8th- and 9th-grade adolescents (N = 1,965) to examine the hypotheses that the prospective relationship between witnessing family violence and directly experiencing violence and physical dating aggression perpetration is mediated by 3 constructs: (a) normative beliefs about dating aggression (norms), (b) anger dysregulation, and (c) depression. Results from cross-lagged regression models suggest that the relationship between having been hit by an adult and dating aggression is mediated by changes in norms and anger dysregulation, but not depression. No evidence of indirect effects from witnessing family violence to dating aggression was found through any of the proposed mediators. Taken together, the findings suggest that anger dysregulation and normative beliefs are potential targets for dating abuse prevention efforts aimed at youth who have directly experienced violence

Severe stress switches CRF action in the nucleus accumbens from appetitive to aversive.

Stressors motivate an array of adaptive responses ranging from \u27fight or flight\u27 to an internal urgency signal facilitating long-term goals. However, traumatic or chronic uncontrollable stress promotes the onset of major depressive disorder, in which acute stressors lose their motivational properties and are perceived as insurmountable impediments. Consequently, stress-induced depression is a debilitating human condition characterized by an affective shift from engagement of the environment to withdrawal. An emerging neurobiological substrate of depression and associated pathology is the nucleus accumbens, a region with the capacity to mediate a diverse range of stress responses by interfacing limbic, cognitive and motor circuitry. Here we report that corticotropin-releasing factor (CRF), a neuropeptide released in response to acute stressors and other arousing environmental stimuli, acts in the nucleus accumbens of naive mice to increase dopamine release through coactivation of the receptors CRFR1 and CRFR2. Remarkably, severe-stress exposure completely abolished this effect without recovery for at least 90 days. This loss of CRF\u27s capacity to regulate dopamine release in the nucleus accumbens is accompanied by a switch in the reaction to CRF from appetitive to aversive, indicating a diametric change in the emotional response to acute stressors. Thus, the current findings offer a biological substrate for the switch in affect which is central to stress-induced depressive disorders