RESUMEN
BACKGROUND: Treatment of low anterior resection syndrome (LARS) is challenging. Percutaneous tibial nerve stimulation (PTNS) can improve select bowel disorders. An RCT was conducted to assess the efficacy of PTNS compared with sham stimulation in patients with severe LARS. METHOD: This was a multicentre, double-blind RCT. Patients with major LARS score were allocated to receive PTNS or sham therapy (needle placement simulation without nerve stimulation). The study included 16 sessions of 30 min once a week for 12 consecutive weeks, followed by four additional sessions once a fortnight for the following 4 weeks. The primary endpoint was efficacy of PTNS defined by the LARS score 12 months after treatment. Secondary endpoints included faecal incontinence, quality of life (QoL), and sexual function. RESULTS: Between September 2016 and July 2018, 46 eligible patients were assigned randomly in a 1 : 1 ratio to PTNS or sham therapy. Baseline characteristics were similar. LARS scores were reduced in both groups, but only patients who received PTNS maintained the effect in the long term (mean(s.d.) score 36.4(3.9) at baseline versus 30.7(11.5) at 12 months; P = 0.018; effect size -5.4, 95 per cent c.i. -9.8 to -1.0), with a mean reduction of 15.7 per cent at 12-month follow-up. The faecal incontinence score was improved after 12 months in the PTNS group (mean(s.d.) score 15.4(5.2) at baseline versus 12.5(6.4) at 12 months; P = 0.018). No major changes in QoL and sexual function were observed in either group. There was no therapy-associated morbidity. Three patients discontinued the study, but none owing to study-related issues. CONCLUSION: PTNS has positive effects in some patients with major LARS, especially in those with faecal incontinence. Registration number: NCT02517853 (http://www.clinicaltrials.gov).
Asunto(s)
Proctocolectomía Restauradora/efectos adversos , Recto/cirugía , Nervio Tibial , Estimulación Eléctrica Transcutánea del Nervio/métodos , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Enfermedades del Recto/etiología , Enfermedades del Recto/terapia , SíndromeRESUMEN
AIMS: Activation of the renin-angiotensin system (RAS), renal oxidative stress and inflammation are constantly present in experimental hypertension. Nitric oxide (NO) inhibition with N(w)-nitro-L-arginine methyl ester (L-NAME) has previously been reported to produce hypertension, increased expression of Angiotensin II (Ang II) and renal dysfunction. The use of Losartan, an Ang II type 1 receptor (AT1R) antagonist has proven to be effective reducing hypertension and renal damage; however, the mechanism by which AT1R blockade reduced kidney injury and normalizes blood pressure in this experimental model is still complete unknown. The current study was designed to test the hypothesis that AT1R activation promotes renal NAD(P)H oxidase up-regulation, oxidative stress and cytokine production during L-NAME induced-hypertension. MAIN METHODS: Male Sprague-Dawley rats were distributed in three groups: L-NAME, receiving 70 mg/100ml of L-NAME, L-NAME+Los, receiving 70 mg/100ml of L-NAME and 40 mg/kg/day of Losartan; and Controls, receiving water instead of L-NAME or L-NAME and Losartan. KEY FINDINGS: After two weeks, L-NAME induced high blood pressure, renal overexpression of AT1R, NAD(P)H oxidase sub-units gp91, p22 and p47, increased levels of oxidative stress, interleukin-6 (IL-6) and interleukin-17 (IL-17). Also, we found increased renal accumulation of lymphocytes and macrophages. Losartan treatment abolished the renal expression of gp91, p22, p47, oxidative stress and reduced NF-κB activation and IL-6 expression. SIGNIFICANCE: These findings indicate that NO induced-hypertension is associated with up-regulation of NADPH oxidase, oxidative stress production and overexpression of key inflammatory mediators. These events are associated with up-regulation of AT1R, as evidenced by their reversal with AT1R blocker treatment.
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Hipertensión/fisiopatología , Inflamación/fisiopatología , Óxido Nítrico/metabolismo , Estrés Oxidativo/fisiología , Receptor de Angiotensina Tipo 1/metabolismo , Bloqueadores del Receptor Tipo 1 de Angiotensina II/farmacología , Animales , Presión Sanguínea/efectos de los fármacos , Hipertensión/tratamiento farmacológico , Inflamación/tratamiento farmacológico , Interleucina-17/metabolismo , Interleucina-6/metabolismo , Losartán/farmacología , Masculino , NADH NADPH Oxidorreductasas/metabolismo , NADPH Oxidasa 4 , FN-kappa B/metabolismo , NG-Nitroarginina Metil Éster/farmacología , Estrés Oxidativo/efectos de los fármacos , Ratas , Ratas Sprague-Dawley , Sistema Renina-Angiotensina/efectos de los fármacos , Sistema Renina-Angiotensina/fisiología , Regulación hacia Arriba/efectos de los fármacosRESUMEN
We examined the interaction between early life stress and vulnerability to alcohol in female rats exposed to prenatal restraint stress (PRS rats). First we studied the impact of PRS on ethanol preference during adolescence. PRS slightly increased ethanol preference per se, but abolished the effect of social isolation on ethanol preference. We then studied the impact of PRS on short- and long-term responses to ethanol focusing on behavioral and neurochemical parameters related to depression/anxiety. PRS or unstressed adolescent female rats received 10% ethanol in the drinking water for 4 weeks from PND30 to PND60. At PND60, the immobility time in the forced-swim test did not differ between PRS and unstressed rats receiving water alone. Ethanol consumption had no effect in unstressed rats, but significantly reduced the immobility time in PRS rats. In contrast, a marked increase in the immobility time was seen after 5 weeks of ethanol withdrawal only in unstressed rats. Hippocampal levels of neuropeptide Y (NPY) and mGlu1a metabotropic glutamate receptors were increased at the end of ethanol treatment only in unstressed rats. Ethanol treatment had no effect on levels of corticotropin-releasing hormone (CRH) in the hippocampus, striatum, and prefrontal cortex of both groups of rats. After ethanol withdrawal, hippocampal levels of mGlu1 receptors were higher in unstressed rats, but lower in PRS rats, whereas NPY and CRH levels were similar in the two groups of rats. These data indicate that early life stress has a strong impact on the vulnerability and responsiveness to ethanol consumption during adolescence.
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Consumo de Bebidas Alcohólicas/fisiopatología , Etanol/farmacología , Efectos Tardíos de la Exposición Prenatal/fisiopatología , Estrés Fisiológico/efectos de los fármacos , Estrés Fisiológico/fisiología , Animales , Conducta de Elección/fisiología , Cuerpo Estriado/metabolismo , Hormona Liberadora de Corticotropina/metabolismo , Femenino , Hipocampo/metabolismo , Pérdida de Tono Postural/efectos de los fármacos , Pérdida de Tono Postural/fisiología , Masculino , Neuropéptido Y/metabolismo , Corteza Prefrontal/metabolismo , Embarazo , Efectos Tardíos de la Exposición Prenatal/psicología , Ratas , Ratas Sprague-Dawley , Receptores de Glutamato Metabotrópico/metabolismo , Aislamiento Social/psicologíaRESUMEN
With the aim to determine the presence of individual nitro-PAH contained in particles in the atmosphere of Mexico City, a monitoring campaign for particulate matter (PM(10) and PM(2.5)) was carried out in Northern Mexico City, from April 2006 to February 2007. The PM(10) annual median concentration was 65.2µgm(-3) associated to 7.6µgm(-3) of solvent-extractable organic matter (SEOM) corresponding to 11.4% of the PM(10) concentration and 38.6µgm(-3) with 5.9µgm(-3) SEOM corresponding to 15.2% for PM(2.5). PM concentration and SEOM varied with the season and the particle size. The quantification of nitro-polycyclic aromatic hydrocarbons (nitro-PAH) was developed through the standards addition method under two schemes: reference standard with and without matrix, the former giving the best results. The recovery percentages varied with the extraction method within the 52 to 97% range depending on each nitro-PAH. The determination of the latter was effected with and without sample purification, also termed fractioning, giving similar results. 8 nitro-PAH were quantified, and their sum ranged from 111 to 819pgm(-3) for PM(10) and from 58 to 383pgm(-3) for PM(2.5), depending on the season. The greatest concentration was for 9-Nitroanthracene in PM(10) and PM(2.5), detected during the cold-dry season, with a median (10th-90th percentiles) concentration in 235pgm(-3) (66-449pgm(-3)) for PM(10) and 73pgm(-3) (18-117pgm(-3)) for PM(2.5). The correlation among mass concentrations of the nitro-PAH and criteria pollutants was statistically significant for some nitro-PAH with PM(10), SEOM in PM(10), SEOM in PM(2.5), NO(X), NO(2) and CO, suggesting either sources, primary or secondary origin. The measured concentrations of nitro-PAH were higher than those reported in other countries, but lower than those from Chinese cities. Knowledge of nitro-PAH atmospheric concentrations can aid during the surveillance of diseases (cardiovascular and cancer risk) associated with these exposures.
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Contaminantes Atmosféricos/análisis , Monitoreo del Ambiente , Compuestos de Nitrógeno/análisis , Material Particulado/análisis , Hidrocarburos Policíclicos Aromáticos/análisis , Contaminación del Aire/estadística & datos numéricos , Atmósfera/química , Ciudades , México , Tamaño de la Partícula , Estaciones del Año , TiempoRESUMEN
A fast preparative two-step chromatographic method for purification of B-phycoerythrin from Porphyridium cruentum is described. This biliprotein was homogeneous as determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis yielding three closely migrating bands corresponding to its three subunits. Baseline separation of its alpha-, beta- and gamma-subunits was achieved by a reversed-phase HPLC gradient semipreparative method with a C4 large-pore column and a solvent system consisting of 0.05% trifluoroacetic acid (TFA) in water and 0.05% TFA in acetonitrile. B-Phycoerythrin in different aggregation states and its subunits have been spectroscopically characterized. Hexameric B-phycoerythrin has similar secondary and tertiary structure than dissociated B-phycoerythrin determined by circular dichroism.
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Cromatografía Líquida de Alta Presión/métodos , Ficoeritrina/aislamiento & purificación , Rhodophyta/química , Electroforesis en Gel de Poliacrilamida , Ficoeritrina/química , Espectrofotometría UltravioletaRESUMEN
Sensitive, safe and easy-to-use probes for the detection of nucleic acids are urgently called for. To this end we are in the process of developing a fluorescence-based technique to work in homogeneous assay media. We have examined pyrene and fluorescein as fluorescent labels for natural DNA probes. A fraction of the cytosine residues of a single-stranded cDNA was randomly labelled with either pyrene or fluorescein using the bisulfite-catalyzed diamine reaction. Both fluorophores showed fluorescence quenching when the labelled probe was hybridized with its complementary strand and we describe the changes in steady-state fluorescence intensity that occurred upon hybridization. Our results demonstrate that pyrene quenching is more efficient than fluorescein quenching and thus pyrene-labelled probes are more sensitive for detecting and quantifying DNA from natural sources.
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ADN/análisis , Fluoresceínas , Colorantes Fluorescentes , Pirenos , Animales , Sondas de ADN , ADN Complementario , Gliceraldehído-3-Fosfato Deshidrogenasas/genética , Hibridación de Ácido Nucleico , Ratas , Sensibilidad y Especificidad , TemperaturaRESUMEN
In the present study, we have evaluated whether melatonin (MEL) modulates Mn-induced decrease in spontaneous motor activity (SMA) and lipid peroxidation, estimated as malondialdehyde (MDA) formation, in several brain regions. In mice treated with manganese a decrease in SMA after 2 weeks of treatment was observed. In the group treated with Mn+MEL a significant greater reduction in SMA was detected at 4 weeks. MDA levels were reduced in both MEL and Mn treated mice. In the animals treated with MEL + Mn a higher reduction in MDA levels was observed. These results suggest that MEL modulates the effect of Mn on SMA and brain lipid peroxidation.
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Encéfalo/metabolismo , Peroxidación de Lípido/efectos de los fármacos , Manganeso/farmacología , Melatonina/farmacología , Actividad Motora/efectos de los fármacos , Animales , Encéfalo/efectos de los fármacos , Interacciones Farmacológicas , Cinética , Malondialdehído/metabolismo , Manganeso/administración & dosificación , Melatonina/administración & dosificación , RatonesRESUMEN
Supplementation of coconut oil to the diet for 1-2 weeks produced a significant hypercholesterolemia in 14-day-old chicks. Changes in plasma fatty acid composition correlated positively with those of diets. In this study, we have shown a different response of low- and high-density lipoprotein (LDL and HDL) fractions to dietary saturated fat (coconut oil) rich in lauric and myristic acids. Although all the components of these particles seemed to increase, the percentages of increases found in total (TC), free (FC) and esterified cholesterol (EC) were higher in LDL than in HDL. TC/phospholipid (PL) ratio, considered as an inverse index of membrane fluidity, also increased with the dietary regimen in LDL, while no significant differences were found in HDL. These results suggest that supplementation of coconut oil to the diet decreased the fluidity of LDL. The EC/triglycerides (TG) ratio was also significantly increased in LDL, corroborating the main atherogenic function of this lipoprotein fraction in response to lauric and myristic acids. We have also estimated the lipidic order parameter, S, from the steady-state fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene (DPH)-labelled low- and high-density lipoproteins. In LDL, temperature dependence of S shows two different behaviour zones at about 20 degrees C. In HDL, the plot of S values versus T is linear. DPH anisotropy and S increased in both LDL and HDL from treated chicks. This increase becomes more evident as temperature rises and also with dietary treatment.
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Cocos , Grasas de la Dieta/administración & dosificación , Lipoproteínas HDL/biosíntesis , Lipoproteínas HDL/química , Lipoproteínas LDL/biosíntesis , Lipoproteínas LDL/química , Aceites de Plantas/administración & dosificación , Animales , Anisotropía , Fenómenos Químicos , Química Física , Pollos , Aceite de Coco , Lipoproteínas/sangre , Lipoproteínas/química , Lipoproteínas HDL/sangre , Lipoproteínas LDL/sangre , Masculino , Plasma/química , TemperaturaRESUMEN
C-phycocyanin and allophycocyanin from the green alga Spirulina platensis were isolated and crystallized by gel-acupuncture techniques. A novel two-step chromatographic procedure was used for purification. Blue hexagonal crystals were obtained by diffusing magnesium chloride into the protein solution for a week, followed by diffusion of PEG 6000 in order to complete the reduction of the solubility of the protein in the capillary tube used as a growth cell. In the case of allophycocyanin, crystals with a size of 0.4 x 0.3 x 0.3 mm were characterized by X-ray diffraction. They belong to space group P6(3)22 with unit-cell parameters a = b = 102.04, c = 131.22 A. The crystals of C-phycocyanin belong to either space group P6 or P6(3) with unit-cell constants a = b = 182.38, c = 60.87 A, alpha = beta = 90, gamma = 120 degrees. The crystals diffract beyond 2.4 and 2.5 A resolution, respectively, using a rotating anode as an X-ray source.
RESUMEN
The aim of this study was to determine the neuronal participation of nitric oxide (NO) in experimental epilepsy. To reach this objective, we established the amount of cells presenting nitric oxide synthase (NOS) and the amygdaline concentrations in the L-arginine-nitric oxide synthesis pathway. A group of fully epileptic rats, induced by the kindling procedure and that had reached at least 10 generalized seizures, was studied. We evaluated behavioral stages, electroencephalographic activities, and histochemical NOS-positive cells and carried out high-pressure liquid chromatography (HPLC) determinations of arginine, citrulline, and glutamic acid. Our results showed that behavioral and electrographic frequency, and duration of epileptic activities, were increased during the kindling process. Image processing system of NOS cells showed two types of intensities in cell stains in hippocampus, caudate-putamen, and amygdala. When we independently counted the two types of NOS stain cells, a selective increase in the number and density of weak-stained cells was observed, while dark-stained cells did not change in the studied structures. Additionally, arginine, citrulline, and glutamic acid concentrations in amygdala increased in kindled animals. The differential and specific increase in the stained cells expressing the nitric oxide synthase, as well as the increase in concentrations of the L-arginine-nitric oxide pathway in amygdala, suggested a relationship with the progressive augmentation in the electrophysiological hyperactivity characteristic of generalized epilepsy.
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Epilepsia Generalizada/fisiopatología , Excitación Neurológica/fisiología , NADPH Deshidrogenasa/metabolismo , Neuronas/enzimología , Óxido Nítrico Sintasa/metabolismo , Animales , Arginina/análisis , Conducta Animal , Encéfalo/citología , Citrulina/análisis , Estimulación Eléctrica , Electrofisiología , Ácido Glutámico/análisis , Histocitoquímica , Procesamiento de Imagen Asistido por Computador , Masculino , Óxido Nítrico/fisiología , Ratas , Ratas WistarRESUMEN
DNA and RNA probes are important analytical reagents in molecular biology and in the detection of infectious and genetic diseases. However, the present polynucleotide probe technology is complex and labor-intensive. We have been investigating the possibility of using fluorescent-labeled DNA probes to develop assays which do not require the separation of free from hybridized probe (homogeneous assays). Such assays are possible if the fluorescence efficiency or fluorescence anisotropy of the fluorescent label changes upon hybridization of probe with target DNA. In this article we examine pyrene as a fluorescent label for DNA or RNA probes. Experiments were performed using a model system in which poly(C) and poly(I) are respectively the probe and target sequences. A small fraction of the nucleotide bases of poly(C) was randomly labeled with pyrene using the bisulfite-catalyzed diamine reaction. The results show that the uncorrected emission spectrum of pyrene-poly(C) decreases by a factor of 4 and shifts toward longer wavelengths upon hybridization with poly(I) at saturating concentrations. The average lifetime changes from 10.78 to 4 ns. These fluorescence changes occur in a wide range of chemical environments, including the high salt concentrations normally used to increase the velocity of the hybridization reaction in clinical assays. The pyrene label can thus be used to readily detect the amount of poly(I) in an unknown sample without having to separate free and bound labeled probe. To unravel the mechanism responsible for the observed changes in fluorescence intensity upon hybridization, we have performed polarized fluorescence intensity measurements and analyzed the results by approximate steady-state expressions that allow evaluation of the relative contributions of changes in lifetimes (fluorescence efficiency) and rotational motions to the changes in fluorescence intensity. The results indicate that the latter changes are due chiefly to changes in lifetime or fluorescence efficiency and that these changes seem to be due to the movement of the pyrene label to a more hydrophilic environment upon hybridization.
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Sondas de ADN , ADN Complementario/genética , Poli C/química , Pirenos/química , Polarización de Fluorescencia , Hibridación de Ácido NucleicoRESUMEN
We determine the opioid peptide content in the rat brain during the ictal phase and postictal depression after pentylenetetrazol kindling rats. Radioimmunoassays with highly specific antisera risen for Met-enkephalin, Leu-enkephalin and octapeptide, were carried out during the ictal phase, and 15, 30 and 60 min after seizures. We always found an initial IR-Met-enkephalin decrease during the postictal depression content, followed by a reduction in IR-Leu-enkephalin and IR-octapeptide tissular concentration. We suggest a functional and differential release of the opioid peptides, during the postictal depression time-course.
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Encéfalo/metabolismo , Endorfinas/metabolismo , Excitación Neurológica , Pentilenotetrazol , Convulsiones/metabolismo , Animales , Encefalina Leucina/metabolismo , Encefalina Metionina/metabolismo , Masculino , Radioinmunoensayo , Ratas , Ratas Wistar , Convulsiones/inducido químicamente , Convulsiones/fisiopatología , Factores de Tiempo , Distribución TisularRESUMEN
The presence and localization of NADPH-diaphorase in the cerebral ganglion of the crayfish Cambarellus montezumae was shown. The reactivity of this enzyme was found in the deuterocerebrum, mainly in the commissure, in fibers of olfactory and accessory lobes, and in the laterodorsal group of cells. The presence of this enzyme in these cerebral regions suggests that nitric oxide is involved in primary sensory afferents in the crayfish.
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Astacoidea/metabolismo , Encéfalo/efectos de los fármacos , NADPH Deshidrogenasa/metabolismo , Animales , Encéfalo/enzimología , Ganglios de Invertebrados/efectos de los fármacos , Ganglios de Invertebrados/enzimología , Neuronas/fisiología , Óxido Nítrico/farmacologíaRESUMEN
Valproic acid (VPA) induces abstinence behavior and analgesia and displays an anticonvulsant effect, but its exact mechanism of action is not yet clear. In order to view whether proenkephalin derived-peptides are involved in the mechanism of VPA-induced behavior, we analyzed immunoreactive-met-enkephalin (IR-ME) in rat striatum, midbrain, and amygdala 10, 20, and 45 min after i.p. injection of 200 mg/kg of VPA. VPA induced body shakes that peaked within 5 to 10 min. IR-ME increased in the striatum and decreased in the midbrain at 10, 20, and 45 min, reaching the highest and lowest levels at 10 and 20 min, respectively. No changes occurred in the amygdala. Gel filtration chromatography followed by HPLC of striatum extracts showed that the increased IR-ME levels corresponded to low molecular weight peptides, including ME. These results indicate that VPA produced rapid changes of IR-ME levels in rat brain and suggest peptide participation in the mechanisms of VPA-induced behavior. The anticonvulsant effect of VPA was tested in rats treated with pentylenetetrazol (70 mg/kg) 30 min after VPA (400 mg/kg) administration, and IR-ME was analyzed in striatum 15 min later. No changes in striatal IR-ME levels occurred in protected rats (no behavioral convulsions), compared with those treated only with VPA, but a significant decrease appeared in unprotected animals (clonic convulsions). These results suggest that striatal ME may participate in the mechanism of VPA-induced abstinence behavior and in the anticonvulsant effect. Otherwise, midbrain ME might be involved in other VPA behaviors such as analgesia.
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Anticonvulsivantes/farmacología , Química Encefálica/efectos de los fármacos , Encefalina Metionina/metabolismo , Síndrome de Abstinencia a Sustancias/psicología , Ácido Valproico/farmacología , Animales , Cromatografía en Gel , Cromatografía Líquida de Alta Presión , Epilepsia Tónico-Clónica/inducido químicamente , Epilepsia Tónico-Clónica/prevención & control , Inyecciones Intraperitoneales , Masculino , Pentilenotetrazol , Ratas , Ratas Wistar , Ácido Valproico/efectos adversosRESUMEN
In this paper we discuss the anatomical localization of NADPH-diaphorase using Nitroblue tetrazolium in perioesophageal ganglia of Helix aspersa. Our results show that the reaction is present in neurons and fibers of the procerebrum, some positive neurons are found in mesocerebrum, and there were positive fibers in the neuropile of postcerebrum and mesocerebrum; likewise, immunopositive fibers were found in the neuropile of pedal, pleural and parietal ganglia. The presence of NADPH-diaphorase in the interneurons of procerebrum suggests the participation of this enzyme in the production of nitric oxide for the processing of the olfactory information, as has been suggested in mammalian olfactory tissue.
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Ganglios de Invertebrados/enzimología , NADPH Deshidrogenasa/metabolismo , Animales , Ganglios de Invertebrados/anatomía & histología , Ganglios de Invertebrados/metabolismo , Caracoles Helix , Histocitoquímica , Interneuronas/enzimología , Interneuronas/fisiología , Óxido Nítrico/biosíntesis , Nitroazul de TetrazolioRESUMEN
This paper shows a medial prefrontal cortex (CxAP9) facilitating influence upon the unit activity of the centralis lateralis (Cl) nucleus of the thalamus, in rats anesthetized with urethane. Cortical influences were studied using both cortical cooling and cortical spreading depression (CSD) procedures. Both spontaneous and noxious thermally evoked activities were considered. When CSD was propagated and affected the CxAP9, as well as during the cooling of this area, both spontaneous activity and the responses evoked in Cl cells by noxious stimulation were blocked. This effect was interpreted as a cortical disfacilitation upon Cl cells. During the cortical silent period we tested the excitability of a few Cl cells, provoking their activation by passing electrical current across the same Cl recording electrode. No changes were observed in their excitable response threshold during CSD or cortical cooling. Our results are in agreement with the proposition of a tonic cortical facilitatory action upon the spontaneous and noxious-evoked responses recorded in the Cl cells.
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Corteza Cerebral/fisiología , Lóbulo Frontal/fisiología , Núcleos Talámicos/fisiología , Animales , Corteza Cerebral/citología , Frío/efectos adversos , Depresión de Propagación Cortical/fisiología , Potenciales Evocados/fisiología , Lóbulo Frontal/citología , Masculino , Ratas , Ratas EndogámicasRESUMEN
Using amygdaloid kindling in chronic rats, we were able to observe behavioral, electrographic and IR-Met- and IR-Leu-enkephalin changes throughout the progress of different stages of convulsive activity. Rats presenting the initial stages of kindling, rats presenting the first generalized motor seizure, and rats with at least 10 generalized seizures were sacrificed 24 h after the last stimulus; also rats with at least 10 generalized seizures but sacrificed 21 days after the last seizure were compared with control and sham-operated groups of rats. The IR-Met and IR-Leu enkephalin concentrations in each group were measured in the striatum, amygdala, hypothalamus, medulla oblongata (including pons), hippocampus, mid-brain, spinal cord and cerebral cortex. A progressive increase in IR-Leu-enkephalin in amygdala and hippocampus was observed over the course of kindling. These increases remained until 21 days after rats were fully kindled (at least 10 generalized seizures). We observed increased and decreased concentration of each peptide in different regions. We discussed the regional and the differential effects of each peptide. The increased concentrations in limbic structures were associated with the amygdaloid increased excitability through the kindling process. We suggest that the decreases in concentrations are related with structures involved in the output behavior manifestations produced by kindling stimulation.
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Amígdala del Cerebelo/metabolismo , Encefalina Leucina/metabolismo , Encefalina Metionina/metabolismo , Excitación Neurológica , Amígdala del Cerebelo/fisiopatología , Animales , Encefalina Leucina/fisiología , Encefalina Metionina/fisiología , Masculino , Radioinmunoensayo , Ratas , Ratas EndogámicasRESUMEN
The peptides met- and leu-enkephalin were identified in the telencephalon, rombencephalon, diencephalon and hypophysis of Ambystoma mexicanum brain by radioimmunoassay procedure. The met-enkephalin was the predominant peptide present in the axolotl brain in contrast with leu-enkephalin, except in the hypophysis where the ratio MET/LEU was 2.2/l. The clear differences in the concentration between enkephalins through a submammalian brain species as Ambystoma genus and the possibility that leu-enkephalin is derived exclusively from a precursor like prodynorphin offers an excellent model for the opioids biosynthetic processes.
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Química Encefálica , Encefalina Leucina/análisis , Encefalina Metionina/análisis , Ambystoma mexicanum , AnimalesRESUMEN
Pentylenetetrazol (PTZ) kindling was induced in male Wistar rats by daily i.p. injections of 40 mg/kg of the convulsant agent. Immunoreactive (IR)-Met-enkephalin was quantified in the amygdala, hippocampus and hypothalamus 17 days after the last stimulus, in groups of 6-7 rats, every 4 h, beginning at 08.00 h. IR-Met-enkephalin level displayed diurnal variations in brain regions of control animals. In the amygdala and the hippocampus the peptide peaked at 24.00 h and in the hypothalamus at 20.00 h; the troughs were at 08.00, 16.00 and 08.00 h, respectively. Diurnal variations were abolished in the amygdala and hypothalamus of kindled rats. In the amygdala the effect was characterized by an IR-Met-enkephalin increase at 04.00, 08.00 and 12.00 h; in the hypothalamus the peptide was enhanced at 08.00 and 12.00 h; in the hippocampus IR-Met-enkephalin increased at 12.00 h and showed a displacement of the peak during the dark phase. The results suggest that PTZ kindling in rats produces a long-lasting alteration on diurnal variations of IR-Met-enkephalin levels in limbic structures.
Asunto(s)
Encéfalo/metabolismo , Ritmo Circadiano , Encefalina Metionina/fisiología , Excitación Neurológica , Amígdala del Cerebelo/metabolismo , Amígdala del Cerebelo/fisiopatología , Animales , Encéfalo/fisiopatología , Cuerpo Estriado/metabolismo , Cuerpo Estriado/fisiopatología , Encefalina Metionina/análisis , Hipocampo/metabolismo , Hipocampo/fisiopatología , Hipotálamo/metabolismo , Hipotálamo/fisiopatología , Masculino , Pentilenotetrazol , Ratas , Ratas EndogámicasRESUMEN
In this work we analyzed the immunoreactive-methionine-enkephalin (IR-Met-enkephalin) levels in several brain regions of rats sacrificed during the tonic extension, induced by acute treatment with pentylenetetrazol (PTZ). The results show an increased of IR-Met-enkephalin content in striatum but not in amygdala, hypothalamus, septum, hippocampus and cortex. To characterize whether this elevation of enkephalin levels in striatum corresponded to the releasable pool, we studied the in vitro efflux of this peptide in striatal slices of rats sacrificed during the seizures, in acute PTZ and in PTZ-kindled rats (kindling group I). In addition, PTZ-kindled rats were analyzed 24 h after the last stimulus (kindling group II). The striatal slices of acute group and kindling group I displayed a significant increase in the evoked release of IR-Met-enkephalin. However, no significant changes occurred from striatal slices of kindling group II animals. In vitro superfusion of GABA (100 microM) produced a approximately equal to 63% decrease in IR-Met-enkephalin released from striatal slices in both saline and acute PTZ-treated rats. Several studies suggest that opioid peptides may be released in the ictal phase of seizure in order to mediate some transient postictal behavior. Our results suggest that of several brain regions tested, only the striatal IR-Met-enkephalin may be released during the ictus to mediate postictal behavior in the acute PTZ treated and in PTZ-kindled rats. This effect may be regulated by the GABA system.