Galanin interacts with Neuropeptide Y Y1 receptor in the dentate gyrus of the hippocampus through GALR2/NPYY1R heterodimers

We have shown Galanin(GAL) and Neuropeptide Y Y1(NPYY1) interactions at behavioural, cellular and receptor levels through GALR2/NPYY1R heterodimers in the amygdala. The aim of this work was to analyze GAL/NPYY1R interactions in the Dentate Gyrus(DG) of the Hippocampus, using autoradiographic, in situ hybridization and in situ proximity ligation assay(PLA). Rats(n=6) were sacrificed 15 minutes or 5 hours after icv injections of GAL(3nmol) and DG sections were incubated with NPYY1R agonist [I125]-[Leu31,Pro34]PYY(25 pM) or NPYY1R-33PdATP specific probe, for autoradiography and in situ hybridization respectively. Autoradiograms were analyzed using NIH image analysis system and Student’s unpaired t-test was used. For PLA, DG sections were incubated with anti-GALR2 Rabbit(1:100) and anti-NPYY1R Goat(1:200). PLA signals were detected with PLA PLUS or MINUS probes for rabbit or goat/mouse antibodies. PLA signals were visualized by using a confocal microscope Leica TCS-SL confocal microscope(Leica). We observed that GAL significant increased the NPYY1R agonist [I125]-[Leu31,Pro34]PYY binding in the DG by 20% (p<0,05) and the NPYY1R mRNA expression in the granular layer of DG by 31% (p<0,001). Moreover, PLA-positive red clusters were found specifically in the polymorphic layer and subgranular zone of the DG. No PLA clusters were observed neither in the molecular layer of the DG nor in the corpus callosum, an area that seems to lack of GALR2 receptor. These results demonstrate a novel mechanism of interaction between GAL and NPY1R in the DG at receptor level, probably involving the formation of GALR2/NPYY1R heteroreceptor complexes. Study supported by Junta de Andalucia CVI6476.

Inequities in energy-balance related behaviours and family environmental determinants in European children: baseline results of the prospective EPHE evaluation study

BACKGROUND:Tackling inequalities in overweight, obesity and related determinants has become a top priority for the European research and policy agendas. Although it has been established that such inequalities accumulate from early childhood onward, they have not been studied extensively in children. The current article discusses the results of an explorative analysis for the identification of inequalities in behaviours and their determinants between groups with high and low socio-economic status. METHODS: This study is part of the Epode for the Promotion of Health Equity (EPHE) evaluation study, the overall aim of which is to assess the impact and sustainability of EPODE methodology to diminish inequalities in childhood obesity and overweight. Seven community-based programmes from different European countries (Belgium, Bulgaria, France, Greece, Portugal, Romania, The Netherlands) participate in the EPHE study. In each of the communities, children aged 6-8 years participated, resulting in a total sample of 1266 children and their families. A parental self-administrated questionnaire was disseminated in order to assess the socio-economic status of the household, selected energy balance-related behaviours (1. fruit and vegetable consumption; 2. soft drink/ fruit juices and water consumption; 3. screen time and 4. sleep duration) of the children and associated family environmental determinants. The Mann-Whitney U test and Pearson's chi-square test were used to test differences between the low and high education groups. The country-specific median was chosen as the cut-off point to determine the educational level, given the different average educational level in every country. RESULTS: Children with mothers of relatively high educational level consumed fruits and vegetables more frequently than their peers of low socio-economic status. The latter group of children had a higher intake of fruit juices and/or soft drinks and had higher screen time. Parental rules and home availability were consistently different between the two socio-economic groups in our study in all countries. However we did not find a common pattern for all behaviours and the variability across the countries was large. CONCLUSIONS: Our findings are indicative of socio-economic inequalities in our samples, although the variability across the countries was large. The effectiveness of interventions aimed at chancing parental rules and behaviour on health inequalities should be studied.

Galanin n-terminal gragment (1-15) modifies the 5-HT1A receptor against [H3]-8-OH-DPAT binding in the dorsal raphe and hippocampus of the rat

We have described that Galanin N-terminal fragment (1-15) [GAL(1-15)] is associated with depressive effects and also modulates the antidepressant effects induced by the 5-HT1A receptor (5-HT1AR) agonist 8-OH-DPAT. The aim of this study is to analyze the ability of GAL(1-15) to modulate 5-HT1AR at the autoreceptor and postsynaptic receptor level in rats by using quantitative autoradiography. We analyzed the effect of intracerebroventricular GAL(1-15)-3nmol (n=6) or aCSF (n=6), 10 minutes, 2 and 5 hours after the injection, on the binding characteristics of the 5-HT1AR agonist [H3]-8-OH-DPAT in sections of the Dorsal Raphe (DR) and Dorsal Hippocampus, specifically CA1 and Dentate Gyrus (DG). Student’s t-test was used to compare the experimental groups. GAL(1-15) produced a time-dependent effect on the binding of [H3]-8-OH-DPAT. In CA1 and DG, a significant increase in the KD and Bmax was observed, by 90%(p<0.05), at 10 minutes and 2 hours after injection. However, 5 hours after GAL(1-15) the only significant change remaining was the increase in Bmax at the DG. The coinjection of the GALR2 antagonist M871 blocked significantly the effects induced by GAL(1-15) in both areas. In DR, 2 hours after injection GAL(1-15) only produced a decrease in the Bmax by 20%(p<0.05). These results indicate that GAL(1-15) interacts with 5-HT1AR at the receptor level in DR and Dorsal Hippocampus. Therapeutic strategies based on these results could be developed for the treatment of depression disorders. This work has been supported by Junta de Andalucia CVI646 and Spanish Ministry of Economy PSI2013-44901-P.

Oligodeoxynucleotide synthesis using protecting groups and a linker cleavable under non-nucleophilic conditions

Oligodeoxynucleotides (ODNs) containing latent electrophilic groups can be highly useful in antisense drug development and many other applications such as chemical biology and medicine, where covalent cross-linking of ODNs with mRNA, protein and ODN is required. However, such ODN analogues cannot be synthesized using traditional technologies due to the strongly nucleophilic conditions used in traditional deprotection/cleavage process. To solve this long lasting and highly challenging problem in nucleic acid chemistry, I used the 1,3-dithian-2-yl-methoxycarbonyl (Dmoc) function to protect the exo-amino groups on the nucleobases dA, dC and dG, and to design the linker between the nascent ODN and solid support. These protecting groups and linker are completely stable under all ODN synthesis conditions, but can be readily cleaved under non-nucleophilic and nearly neutral conditions. As a result, the new ODN synthesis technology is universally useful for the synthesis of electrophilic ODNs. The dissertation is mainly comprised of two portions. In the first portion, the development of the Dmoc-based linker for ODN synthesis will be described. The construction of the dT-Dmoc-linker required a total of seven steps to synthesize. The linker was then anchored to the solid support―controlled pore glass (CPG). In the second portion, the syntheses of Dmoc-protected phosphoramidites ODN synthesis monomers including Dmoc-dC-amidite, Dmoc-dA-amidite, Dmoc-dG-amidite are described. The protection of dC and dA with 1,3-dithian-2-yl-methyl 4-nitrophenyl carbonate proceeded smoothly giving Dmoc-dC and Dmoc-dA in good yields. However, when the same acylation procedure was applied for the synthesis of Dmoc-dG, very low yield was obtained. This problem was later solved using a highly innovative and environmentally benign procedure, which is expected to be widely useful for the acylation of the exo-amino groups on nucleoside bases. The reactions to convert the Dmoc-protected nucleosides to phosphoramidite monomers proceeded smoothly with high yields. Using the Dmoc phosphoramidite monomers dA, dC, dG and the commercially available dT, and the Dmoc linker, four ODN sequences were synthesized. In all cases, excellent coupling yields were obtained. ODN deprotection/cleavage was achieved by using non-nucleophilic oxidative conditions. The new technology is predicted to be universally useful for the synthesis of ODNs containing one or more electrophilic functionalities.

Co-Composting of Poultry Manure with Different Organic Amendments

To study the influence of different organic amendments on the quality of poultry manure compost, three pilot composting trials were carried out with different mixes: poultrymanure/carcasse meal/ashes/grape pomace (Pile 1), poultry manure/cellulosic sludge (Pile 2) and poultry manure (Pile 3). For all piles, wood chips were applied as bulking agent. The process was monitored, over time, by evaluating standard physical and chemical parameters, such as, pH, electric conductivity, moisture, organic matter and ash content, total carbon and total nitrogen content, carbon/nitrogen ratio (C/N) and content in mineral elements. Piles 1 and 2 reached a thermophilic phase, however having different trends. Pile 1 reached this phase earlier than Pile 2. For both, the pH showed a slight alkaline character and the electric conductivity was lower than 2 mS/cm. Also, the initial C/N value was 22 and reached values lower than 15 at the end of composting process. The total N content of the Pile 1 increased slightly during composting, in contrast with the others piles. At the end of composting process, the phosphorus content ranged between 54 and 236 mg/kg dry matter, for Pile 2 and 3, respectively. Generally, the Piles 1 and 3 exhibited similar heavy metals content. This study showed that organic amendments can be used as carbon source, given that the final composts presented parameters within the range of those recommended in the 2nd Draft of EU regulation proposal (DG Env.A.2 2001) for compost quality.

Impedancias basales registradas durante la denervación renal por radiofrecuencia en pacientes con hipertensión arterial resistente

Objetivo: Determinar diferencias en las impedancias basales registradas durante los procedimientos de denervación renal por radiofrecuencia de los pacientes sometidos a este procedimiento en la Fundación Cardioinfantil de Bogotá durante los años 2012 a 2015. Materiales y métodos: Estudio observacional, analítico de corte retrospectivo, donde se analizaron todas las impedancias basales medidas durante los procedimientos de denervación renal, buscando diferencias significativas entre los segmentos de las arterias intervenidas, estratificados en proximal, medio distal y superior, lateral, inferior u ostial. Con seguimiento a los pacientes a tres, seis y doce meses en cuanto a presión arterial de consultorio. Resultados: Se evaluaron 150 puntos de denervación renal exitosos, correspondientes a 23 arterias renales de 11 procedimientos. La mediana de edad fue 56 años. Al realizar un modelo de regresión lineal no se encontró ninguna diferencia estadísticamente significativa entre las impedancias de ninguno de los segmentos de las arterias ni sitios anatómicos. Se documentó disminución de presión arterial sistólica a tres meses, seis meses y doce meses de 14 mmHg (RIQ 10-33mmHg), 21 mmHg (RIQ 12-42mmHg) y 19 mmHg (RIQ 11-42 mmHg) respectivamente

Efecto de los dispositivos de emisión de microondas, radiofrecuencia no ablativa y ultrasonido microfocalizado en el tratamiento de la hiperhidrosis primaria revisión sistemática de la literatura

Antecedentes y objetivos: La hiperhidrosis primaria afecta el 2,8% de la población de Estados Unidos. Condición que impacta el desarrollo social de los individuos afectados, ocasionando fobia social. Existen opciones disponibles para el tratamiento de la hiperhidrosis incluyendo medicamentos tópico, sistémico, inyectable y quirúrgico. El objetivo de ésta revisión sistemática de la literatura es determinar la efectividad y seguridad de los dispositivos de emisión de microondas, radiofrecuencia no ablativa y sistema de ultrasonido microfocalizado para el tratamiento de la hiperhidrosis primaria. Materiales y métodos: Se realizó una revisión sistemática de la literatura de artículos obtenidos de bases de datos: Medline, Cochrane, Embase, Ovid y Scielo. Se incluyeron ensayos clínicos aleatorizados controlados, ensayos cuasiexperimentales desde el 2011; donde evaluaran el uso de estos dispositivos en el manejo de hiperhidrosis primaria. Resultados: Se seleccionaron 21 artículos en total. Se encontró que con los tres dispositivos se logra una reducción significativa a puntajes entre 1 y 2 de la escala de Severidad de la Hiperhidrosis; en 3 estudios se encontró mejoría en la calidad de vida; los eventos adversos fueron transitorios, siendo más frecuentes con el dispositivo de emisión de microondas. Conclusión: Primera revisión sistemática de la literatura sobre el efecto de estos tres dispositivos en el manejo de hiperhidrosis. Se espera aportar a la literatura existente una recomendación acerca de la efectividad y seguridad de estos dispositivos para que sea aplicado en los pacientes con diagnóstico de hiperhidrosis primaria.