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Microbiota-modulating strategies, including probiotic administration, have been tested for the treatment of chronic gastrointestinal diseases despite limited information regarding their mechanisms of action. We previously demonstrated that patients with active celiac disease have decreased duodenal expression of elafin, a human serine protease inhibitor, and supplementation of elafin by a recombinant Lactococcus lactis strain prevents gliadin-induced immunopathology in the NOD/DQ8 mouse model of gluten sensitivity. The commensal probiotic strain Bifidobacterium longum NCC2705 produces a serine protease inhibitor (Srp) that exhibits immune-modulating properties. Here, we demonstrate that B. longum NCC2705, but not a srp knockout mutant, attenuates gliadin-induced immunopathology and impacts intestinal microbial composition in NOD/DQ8 mice. Our results highlight the beneficial effects of a serine protease inhibitor produced by commensal B. longum strains.IMPORTANCE Probiotic therapies have been widely used to treat gastrointestinal disorders with variable success and poor mechanistic insight. Delivery of specific anti-inflammatory molecules has been limited to the use of genetically modified organisms, which has raised some public and regulatory concerns. By examining a specific microbial product naturally expressed by a commensal bacterial strain, we provide insight into a mechanistic basis for the use of B. longum NCC2705 to help treat gluten-related disorders.
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OBJECTIVE: As an initiative of the German Pension Insurance Association (DRV), evidence-based therapeutic modules (ETM) for the rehabilitation of patients with depression were developed. The objective of the subsequent analysis was to analyse the therapeutic procedures in inpatient rehabilitation on the basis of the ETM to evaluate the principal needs for therapeutic standards. METHODS: Data based on the German Classification of Therapeutic Procedures (KTL) for 21 529 patients treated in rehabilitation clinics for people with mental illnesses was analysed with respect to differences between diagnostic groups/clinics regarding type, quantity and duration of measures coded. RESULTS: The mean quantity and duration of the interventions for patients with depressive disorders encoded varied greatly between the ETM. No or only minor differences were found between patients with depression and those with other diagnoses regarding the type, quantity and duration of measures coded. However, there were great variances between clinics. CONCLUSIONS: Therapeutic standards for rehabilitative practice appear necessary in order to reduce treatment heterogeneity between rehabilitation facilities, which could improve the quality of healthcare.
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Trastorno Depresivo/rehabilitación , Adhesión a Directriz/estadística & datos numéricos , Hospitalización/estadística & datos numéricos , Guías de Práctica Clínica como Asunto , Centros de Rehabilitación/normas , Rehabilitación/clasificación , Rehabilitación/normas , Adolescente , Trastorno Depresivo/epidemiología , Medicina Basada en la Evidencia , Femenino , Alemania/epidemiología , Humanos , Masculino , Persona de Mediana Edad , Adulto JovenRESUMEN
Type 2 diabetes is a polygenic and genetically heterogeneous disease . The age of onset of the disease is usually late and environmental factors may be required to induce the complete diabetic phenotype. Susceptibility genes for diabetes have not yet been identified. Islet-brain-1 (IB1, encoded by MAPK8IP1), a novel DNA-binding transactivator of the glucose transporter GLUT2 (encoded by SLC2A2), is the homologue of the c-Jun amino-terminal kinase-interacting protein-1 (JIP-1; refs 2-5). We evaluated the role of IBi in beta-cells by expression of a MAPK8IP1 antisense RNA in a stable insulinoma beta-cell line. A 38% decrease in IB1 protein content resulted in a 49% and a 41% reduction in SLC2A2 and INS (encoding insulin) mRNA expression, respectively. In addition, we detected MAPK8IP1 transcripts and IBi protein in human pancreatic islets. These data establish MAPK8IP1 as a candidate gene for human diabetes. Sibpair analyses performed on i49 multiplex French families with type 2 diabetes excluded MAPK8IP1 as a major diabetogenic locus. We did, however, identify in one family a missense mutation located in the coding region of MAPK8IP1 (559N) that segregated with diabetes. In vitro, this mutation was associated with an inability of IB1 to prevent apoptosis induced by MAPK/ERK kinase kinase 1 (MEKK1) and a reduced ability to counteract the inhibitory action of the activated c-JUN amino-terminal kinase (JNK) pathway on INS transcriptional activity. Identification of this novel non-maturity onset diabetes of the young (MODY) form of diabetes demonstrates that IB1 is a key regulator of 3-cell function.
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Proteínas Adaptadoras Transductoras de Señales , Diabetes Mellitus Tipo 2/genética , Islotes Pancreáticos/metabolismo , Proteínas Nucleares/genética , Transactivadores/genética , Edad de Inicio , Apoptosis/genética , Ensayo de Unidades Formadoras de Colonias , Diabetes Mellitus Tipo 2/epidemiología , Femenino , Efecto Fundador , Francia/epidemiología , Predisposición Genética a la Enfermedad , Genotipo , Transportador de Glucosa de Tipo 2 , Humanos , Insulina/metabolismo , Secreción de Insulina , Insulinoma/genética , Insulinoma/metabolismo , Insulinoma/patología , Proteínas Quinasas JNK Activadas por Mitógenos , Escala de Lod , Sistema de Señalización de MAP Quinasas , Masculino , Proteínas Quinasas Activadas por Mitógenos/fisiología , Proteínas de Transporte de Monosacáridos/metabolismo , Proteínas Nucleares/fisiología , Obesidad/genética , Neoplasias Pancreáticas/genética , Neoplasias Pancreáticas/metabolismo , Neoplasias Pancreáticas/patología , Linaje , Transactivadores/fisiología , Transcripción Genética , Células Tumorales Cultivadas/metabolismoRESUMEN
Epithelioid trophoblastic tumor (ETT) is a rare gestational trophoblastic tumor, first described by Shih and Kurman in 1998. ETT often present as abnormal vaginal bleeding in women of reproductive age, but unlike more common forms of GTN tend to produce much less human chorionic gonadotropin (hCG) for the volume of disease present. ETT can occur after any gestational event and can occur in both intrauterine and extrauterine sites. We present a case of a 46-year-old female patient incidentally diagnosed with ETT and hepatic metastasis. Therapy was multimodal and involved chemotherapy, operation, thermoablation of liver metastases and immunocheckpoint inhibitor. The patient remains disease free for almost four years now. ETT presents a diagnostic challenge due to their rarity and histologic resemblance to other pathologies. ETT can be relatively chemo resistant and are therefore often treated surgically. Misdiagnosis might delay effective treatment and affects survival.
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The occurrence and fluxes of 18 priority substances and emerging pollutants listed in the European Union Water Framework Directive and a Watch List (trace metals (Cd, Pb and Ni), nonylphenols, octylphenols, 8 polyaromatic hydrocarbons, 4 dioxin-like polychlorinated biphenyls and diclofenac) were investigated in a Ukrainian city and the mass discharge loads of these compounds into EU-transboundary watersheds were estimated. Fluxes of chemicals were calculated per capita and per area of the Ukrainian urban territory and used to estimate mass loading of priority and emerging concern compounds from Lviv, Uzhorod and Chernivtsi (West Ukraine) to neighbouring EU-transboundary rivers. The highest loading was found for trace metals (1.15â¯tâ¯a-1), diclofenac (0.7â¯tâ¯a-1) and nonylphenols (0.4â¯tâ¯a-1). Transboundary water contamination must be considered in order to successfully manage water resources in a manner that fulfils the requirements of EU environmental quality standards.
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Monitoreo del Ambiente , Ríos/química , Contaminantes Químicos del Agua/análisis , Diclofenaco/análisis , Fenoles/análisis , Bifenilos Policlorados/análisis , UcraniaRESUMEN
Islet-brain 1 (IB1) is the human and rat homologue of JIP-1, a scaffold protein interacting with the c-Jun amino-terminal kinase (JNK). IB1 expression is mostly restricted to the endocrine pancreas and to the central nervous system. Herein, we explored the transcriptional mechanism responsible for this preferential islet and neuronal expression of IB1. A 731-bp fragment of the 5' regulatory region of the human MAPK8IP1 gene was isolated from a human BAC library and cloned upstream of a luciferase reporter gene. This construct drove high transcriptional activity in both insulin-secreting and neuron-like cells but not in unrelated cell lines. Sequence analysis of this promoter region revealed the presence of a neuron-restrictive silencer element (NRSE) known to bind repressor zinc finger protein REST. This factor is not expressed in insulin-secreting and neuron-like cells. By mobility shift assay, we confirmed that REST binds to the NRSE present in the IB1 promoter. Once transiently transfected in beta-cell lines, the expression vector encoding REST repressed IB1 transcriptional activity. The introduction of a mutated NRSE in the 5' regulating region of the IB1 gene abolished the repression activity driven by REST in insulin-secreting beta cells and relieved the low transcriptional activity of IB1 observed in unrelated cells. Moreover, transfection in non-beta and nonneuronal cell lines of an expression vector encoding REST lacking its transcriptional repression domain relieved IB1 promoter activity. Last, the REST-mediated repression of IB1 could be abolished by trichostatin A, indicating that deacetylase activity is required to allow REST repression. Taken together, these data establish a critical role for REST in the control of the tissue-specific expression of the human IB1 gene.
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Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/biosíntesis , Proteínas Portadoras/genética , Regulación Enzimológica de la Expresión Génica , Proteínas Represoras/metabolismo , Proteínas Represoras/fisiología , Factores de Transcripción/metabolismo , Factores de Transcripción/fisiología , Células 3T3 , Animales , Secuencia de Bases , Northern Blotting , Proteínas Portadoras/metabolismo , ADN Complementario/metabolismo , Inhibidores Enzimáticos/farmacología , Biblioteca de Genes , Células HeLa , Humanos , Ácidos Hidroxámicos/farmacología , Ratones , Datos de Secuencia Molecular , Mutación , Neuronas/metabolismo , Células PC12 , Reacción en Cadena de la Polimerasa , Regiones Promotoras Genéticas , ARN Mensajero/metabolismo , Ratas , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transcripción Genética , Activación Transcripcional , Transfección , Células Tumorales Cultivadas , Dedos de ZincRESUMEN
Mentally retarded people usually receive care in special social, vocational, behavioral or educational facilities. Only recently, we gained some experience in the rehabilitation after trauma of those with mental retardation. We presume that with the increasing awareness of the benefits of comprehensive and early rehabilitation after trauma, orthopedic surgeons, neurosurgeons, and physicians who work in intensive care units, refer more patients who had never before gained from this specialty. We would like to share our experience of the unique rehabilitation process of this population.
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Discapacidad Intelectual/complicaciones , Rehabilitación/métodos , Heridas y Lesiones/complicaciones , Heridas y Lesiones/rehabilitación , Adulto , Femenino , Humanos , Masculino , Persona de Mediana EdadRESUMEN
Infections with arthropod-borne pathogens are an increasing threat world-wide that requires heightened vigilance from veterinary and medical practitioners, especially when they involve new or unusual organisms. A dog was presented to a local veterinary clinic in Germany with malaise, pale mucous membranes and stiff joints. Clinical assessment revealed pyrexia, leukopenia and thrombocytopenia. On suspicion of a tick-borne infection, blood samples were examined for clinical and biochemical parameters and subjected to a Anaplasma phagocytophilum-, Borrelia spp.- and Ehrlichia canis-specific real-time PCR. Additionally, a sample of the pre-therapeutic buffy coat was co-cultured with the Ixodes scapularis cell-line ISE6 for 20days. Only the PCR specific for A. phagocytophilum DNA yielded a positive result, and furthermore, Anaplasma morulae were visible in granulocytes and tick cells. After co-culturing, extracellular trypomastigote and epimastigote stages of Trypanosoma sp. with an average length of 29.7µm were observed, featuring a pointed posterior end. Sequence analysis of a 2080bp fragment of the 18S rRNA gene showed 99% identity to the 18S rRNA gene of Trypanosoma pestanai, previously described from a European badger (Meles meles) in France. The dog's condition improved rapidly in response to doxycycline treatment for three weeks. The clinical status normalized and clinical blood parameters were found to be within the reference ranges. To our knowledge this is the first description of T. pestanai infection in a dog, the first detection of T. pestanai in Germany and the first documented co-infection with these two pathogens. Co-infections with unusual opportunistic vector-borne pathogens should be considered, if acute canine granulocytic anaplasmosis is evident.
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BACKGROUND: Poor performance is often suspected to be associated with EIPH in barrel racing horses; however, there are no published reports of EIPH for this discipline. The prevalence of EIPH in barrel racing horses is also unknown. OBJECTIVES: This study was performed to determine the prevalence of EIPH and signs of airway inflammation in barrel racing horses under normal racing conditions in Alberta. ANIMALS: About 170 barrel racing horses. METHODS: Observational cross-sectional study. Tracheobronchoscopic examinations were performed at least 30 minutes postrace. Video recordings were scored off-site independently by two observers for EIPH and tracheal mucus accumulation (TMA). Horses with an EIPH score ≥2 were not assessed for TMA. Interobserver agreement was calculated by weighted κ statistics. Run times, environmental variables, and clinical information were also recorded for analysis. RESULTS: 77/170 (45.3%) of horses examined showed evidence of EIPH (grade ≥ 1). Interobserver agreement was 0.94. 140/141 (99.3%) of horses assessed for TMA showed evidence of tracheal mucus accumulation (grade ≥ 1) with 104/141 (73.8%) having a TMA score ≥ 2. Interobserver agreement was 0.73. A weak positive association was found between EIPH scores and average run speed, the presence of cough at rest reported by the riders, increased recovery time, exercise intolerance, and outdoor pattern. CONCLUSIONS AND CLINICAL IMPORTANCE: The high prevalence of EIPH observed in the sampled population indicates that barrel racing induces substantial stress on the lungs. The presence of EIPH did not impact negatively on performance. Factors such as environmental dust and frequent traveling might have contributed to the high prevalence of TMA observed.
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Broncoscopía/veterinaria , Hemorragia/veterinaria , Enfermedades de los Caballos/diagnóstico , Inflamación/veterinaria , Enfermedades Pulmonares/veterinaria , Animales , Estudios Transversales , Femenino , Hemorragia/diagnóstico , Hemorragia/etiología , Enfermedades de los Caballos/patología , Caballos , Inflamación/diagnóstico , Inflamación/etiología , Enfermedades Pulmonares/diagnóstico , Enfermedades Pulmonares/etiología , Masculino , Condicionamiento Físico AnimalRESUMEN
Macrophage migration-inhibitory factor (MIF) has recently been identified as a pituitary hormone that functions as a counterregulatory modulator of glucocorticoid action within the immune system. In the anterior pituitary gland, MIF is expressed in TSH- and ACTH-producing cells, and its secretion is induced by CRF. To investigate MIF function and regulation within pituitary cells, we initiated the characterization of the MIF 5'-regulatory region of the gene. The -1033 to +63 bp of the murine MIF promoter was cloned 5' to a luciferase reporter gene and transiently transfected into freshly isolated rat anterior pituitary cells. This construct drove high basal transcriptional activity that was further enhanced after stimulation with CRF or with an activator of adenylate cyclase. These transcriptional effects were associated with a concomitant rise in ACTH secretion in the transfected cells and by an increase in MIF gene expression as assessed by Northern blot analysis. A cAMP-responsive element (CRE) was identified within the MIF promoter region which, once mutated, abolished the cAMP responsiveness of the gene. Using this newly identified CRE, DNA-binding activity was detected by gel retardation assay in nuclear extracts prepared from isolated anterior pituitary cells and AtT-20 corticotrope tumor cells. Supershift experiments using antibodies against the CRE-binding protein CREB, together with competition assays and the use of recombinant CREB, allowed the detection of CREB-binding activity with the identified MIF CRE. These data demonstrate that CREB is the mediator of the CRF-induced MIF gene transcription in pituitary cells through an identified CRE in the proximal region of the MIF promoter.
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Hormona Liberadora de Corticotropina/fisiología , Proteína de Unión a Elemento de Respuesta al AMP Cíclico/fisiología , Factores Inhibidores de la Migración de Macrófagos/genética , Adenohipófisis/fisiología , Activación Transcripcional/genética , Animales , Línea Celular , AMP Cíclico/fisiología , Factores Inhibidores de la Migración de Macrófagos/fisiología , Ratones , Adenohipófisis/citología , Adenohipófisis/metabolismo , Regiones Promotoras Genéticas/efectos de los fármacos , Unión Proteica/genética , RatasRESUMEN
Synaptic activation of gamma-aminobutyric acid (GABA)B receptors at GABA synapses causes (a) postsynaptic hyperpolarization mediating a slow inhibitory postsynaptic potential/current (IPSP/C) and (b) presynaptic inhibition of GABA release which depresses IPSPs and leads to paired-pulse widening of excitatory postsynaptic potentials (EPSPs). To address whether these effects are mediated by pharmacologically identical receptors the effects of six GABA(B) receptor antagonists of widely ranging potencies were tested against each response. Monosynaptic IPSP(B)s were recorded in the presence of GABA(A), AMPA/kainate and NMDA receptor antagonists. All GABA(B) receptor antagonists tested depressed the IPSP(B) with an IC50 based rank order of potency of CGP55679> or =CGP56433 = CGP55845A = CGP52432>CGP51176>CGP36742. Paired-pulse EPSP widening was recorded as an index of paired-pulse depression of GABA-mediated IPSP/Cs. A similar rank order of potency of antagonism of paired-pulse widening was observed to that for IPSP(B) inhibition. Comparison of the IC50 values for IPSP(B) inhibition and paired-pulse EPSP widening revealed a close correlation between the two effects in that their IC50s lay within the 95% confidence limits of a correlation line that described IC50 values for inhibition of paired-pulse EPSP widening that were 7.3 times higher than those for IPSP(B) inhibition. Using the compounds tested here it is not possible to assign different subtypes of GABA(B) receptor to pre- and post-synaptic loci at GABAergic synapses. However, 5-10 fold higher concentrations of antagonist are required to block presynaptic as opposed to postsynaptic receptors when these are activated by synaptically released GABA.
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Antagonistas de Receptores de GABA-B , Hipocampo/fisiología , Receptores Presinapticos/antagonistas & inhibidores , Sinapsis/fisiología , Ácido gamma-Aminobutírico/fisiología , Animales , Autorreceptores/antagonistas & inhibidores , Electrofisiología , Potenciales Postsinápticos Excitadores/fisiología , Antagonistas del GABA/farmacología , Hipocampo/efectos de los fármacos , Técnicas In Vitro , Microelectrodos , Ratas , Ratas Wistar , Receptores AMPA/antagonistas & inhibidores , Receptores de Ácido Kaínico/antagonistas & inhibidoresRESUMEN
1. The pharmacological properties of CGP 37849 (DL-(E)-2-amino-4-methyl-5-phosphono-3-pentenoic acid; 4-methyl-APPA) and its carboxyethylester, CGP 39551, novel unsaturated analogues of the N-methyl-D-aspartate (NMDA) receptor antagonist, 2-amino-5-phosphonopentanoate (AP5), were evaluated in rodent brain in vitro and in vivo. 2. Radioligand binding experiments demonstrated that CGP 37849 potently (Ki 220 nM) and competitively inhibited NMDA-sensitive L-[3H]-glutamate binding to postsynaptic density (PSD) fractions from rat brain. It inhibited the binding of the selective NMDA receptor antagonist, [3H]-((+/-)-3-(2-carboxypiperazin-4-yl)propyl-1-phosphonate (CPP), with a Ki of 35 nM, and was 4, 5 and 7 fold more potent than the antagonists [+/-)-cis-4-phosphonomethylpiperidine-2-carboxylic acid) (CGS 19755), CPP and D-AP5, respectively. Inhibitory activity was associated exclusively with the trans configuration of the APPA molecule and with the D-stereoisomer. CGP 39551 showed weaker activity at NMDA receptor recognition sites and both compounds were weak or inactive at 18 other receptor binding sites. 3. CGP 37849 and CGP 39551 were inactive as inhibitors of L-[3H]-glutamate uptake into rat brain synaptosomes and had no effect on the release of endogenous glutamate from rat hippocampal slices evoked by electrical field stimulation. 4. In the hippocampal slice in vitro, CGP 37849 selectively and reversibly antagonized NMDA-evoked increases in CA1 pyramidal cell firing rate. In slices bathed in medium containing low Mg2+ levels, concentrations of CGP 37849 up to 10 microM suppressed burst firing evoked in CAl neurones by stimulation of Schaffer collateral-commissural fibres without affecting the magnitude of the initial population spike; CGP 39551 exerted the same effect but was weaker. In vivo, oral administration to rats of either CGP 37849 or CGP 39551 selectively blocked firing in hippocampal neurones induced by ionophoreticallyapplied NMDA, without affecting the responses to quisqualate or kainate. 5. CGP 37849 and CGP 39551 suppressed maximal electroshock-induced seizures in mice with ED50 s of 21 and 4 mg kg'- p.o., respectively. 6. CGP 37849 and CGP 39551 are potent and competitive NMDA receptor antagonists which show significant central effects following oral administration to animals. As such, they may find value as tools to elucidate the roles of NMDA receptors in brain function, and potentially as therapeutic agents for the treatment of neurological disorders such as epilepsy and ischaemic brain damage in man.
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2-Amino-5-fosfonovalerato/análogos & derivados , Receptores de Neurotransmisores/antagonistas & inhibidores , 2-Amino-5-fosfonovalerato/farmacología , Animales , Anticonvulsivantes , Unión Competitiva/efectos de los fármacos , Electrofisiología , Electrochoque , Glutamatos/metabolismo , Hipocampo/efectos de los fármacos , Hipocampo/metabolismo , Técnicas In Vitro , Cinética , Masculino , Neurotransmisores/metabolismo , Ensayo de Unión Radioligante , Ratas , Ratas Endogámicas , Receptores de N-Metil-D-Aspartato , Receptores de Neurotransmisores/metabolismo , Sodio/metabolismo , Valina/análogos & derivados , Valina/farmacologíaRESUMEN
The high Km glucose transporter GLUT2 is a membrane protein expressed in tissues involved in maintaining glucose homeostasis, and in cells where glucose-sensing is necessary. In many experimental models of diabetes, GLUT2 gene expression is decreased in pancreatic beta-cells, which could lead to a loss of glucose-induced insulin secretion. In order to identify factors involved in pancreatic beta-cell specific expression of GLUT2, we have recently cloned the murine GLUT2 promoter and identified cis-elements within the 338-bp of the proximal promoter capable of binding islet-specific trans-acting factors. Furthermore, in transient transfection studies, this 338-bp fragment could efficiently drive the expression of the chloramphenicol acetyl transferase (CAT) gene in cell lines derived from the endocrine pancreas, but displayed no promoter activity in non-pancreatic cells. In this report, we tested the cell-specific expression of a CAT reporter gene driven by a short (338 bp) and a larger (1311 bp) fragment of the GLUT2 promoter in transgenic mice. We generated ten transgenic lines that integrated one of the constructs. CAT mRNA expression in transgenic tissues was assessed using the RNAse protection assay and the quantitative reverse transcribed polymerase chain reaction (RT-PCR). Overall CAT mRNA expression for both constructs was low compared to endogenous GLUT2 mRNA levels but the reporter transcript could be detected in all animals in the pancreatic islets and the liver, and in a few transgenic lines in the kidney and the small intestine. The CAT protein was also present in Langerhans islets and in the liver for both constructs by immunocytochemistry. These findings suggest that the proximal 338 bp of the murine GLUT2 promoter contain cis-elements required for the islet-specific expression of GLUT2.
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Islotes Pancreáticos/metabolismo , Proteínas de Transporte de Monosacáridos/genética , Regiones Promotoras Genéticas , Animales , Secuencia de Bases , Cloranfenicol O-Acetiltransferasa/genética , Cloranfenicol O-Acetiltransferasa/metabolismo , Clonación Molecular , Cartilla de ADN/genética , Expresión Génica , Genes Reporteros , Transportador de Glucosa de Tipo 2 , Inmunohistoquímica , Hígado/metabolismo , Ratones , Ratones Transgénicos , Datos de Secuencia Molecular , ARN Mensajero/genética , ARN Mensajero/metabolismoRESUMEN
The knowledge on the neuronal inputs to the locus coeruleus (LC) and their roles in regulating noradrenergic (NA) cellular activity is quite advanced. In recent years, however, about ten neuropeptides were found to be localized in the area of the rodent LC; peptides which may be considered as potential transmitters or modulators acting in this area. Electrophysiological studies performed in vivo and in vitro have revealed that many of these peptides are able to alter LC neuronal activity. Stimulatory effects have been described with vasopressin, substance P, adrenocorticotropin hormone and corticotropin-releasing factor. Depressant effects were seen with galanin, somatostatin, neuropeptide Y and enkephalin. Variable actions were observed in the case of neurotensin. While these findings point to a possible regulatory function of these peptides in this area, precise roles remain unclear. Important information is lacking that would conclusively demonstrate their regulatory functions. It should be determined whether the stimulation of peptidergic cells elicits synaptic effects identical to the ones observed with local exogenous peptide applications. By studying the action of blockers of these transmitter and modulator candidates, we would probably begin to understand their importance in the regulation of tonic and phasic activity components. The LC is generally considered to consist of a homogenous group of neurons. The recent observation that subpopulations of these cells contain peptides as in the case of neuropeptide Y, galanin and vasopressin, points to the possible existence of subgroups of neurons having different functions.
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Locus Coeruleus/efectos de los fármacos , Neuropéptidos/farmacología , Animales , Electrofisiología , Locus Coeruleus/fisiología , Potenciales de la Membrana/efectos de los fármacos , Neuronas/efectos de los fármacos , Neuronas/fisiología , Neuropéptidos/fisiología , RoedoresRESUMEN
Stage II colorectal carcinoma is characterized by negative lymph node pathology as determined by conventional microscopic examination. These patients generally do not receive adjuvant therapy although 20%-30% will die from metastatic disease. To determine whether K-ras mutations at codon 12 could be used as a sensitive indicator of occult lymph node metastasis in stage II colon carcinoma, a retrospective study was performed using restriction endonuclease-mediated selective polymerase chain reaction (REMS-PCR) amplification. Of 106 colonic tumors analyzed, 46 were identified as positive for a K12-ras mutation in the primary tumor. Multiple lymph node samples from 38 of these 46 patients were examined by a sensitive nested PCR protocol for the presence of a K12-ras mutation. Of these 38 patients, 14 had 1 or more positive lymph nodes by PCR (37%) and 24 were negative for the mutation (63%). Of the 14 patients with a K12-ras mutation detected in lymph nodes, 8 died of the disease within 5 years (57%) compared to only 4 of the 24 patients with ras-negative lymph nodes (17%). The difference in time to death from disease, stratified using K12-ras status of lymph nodes, was statistically significant (P = 0.036; log-rank test). These results suggest K-ras mutation status of lymph nodes in patients with stage II colon cancer might identify a subgroup of patients who are more likely to develop recurrent and/or metastatic disease and benefit from adjuvant therapy. Larger studies are indicated to determine whether detection of K-ras mutation positivity in histologically negative lymph nodes portends a poor prognosis and to determine whether more aggressive use of adjuvant therapy is warranted.
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Adenocarcinoma Mucinoso/epidemiología , Adenocarcinoma Mucinoso/genética , Adenocarcinoma/epidemiología , Adenocarcinoma/genética , Neoplasias Colorrectales/epidemiología , Neoplasias Colorrectales/genética , Genes ras/genética , Mutación/genética , Adenocarcinoma/diagnóstico , Adenocarcinoma Mucinoso/diagnóstico , Neoplasias Colorrectales/diagnóstico , Supervivencia sin Enfermedad , Humanos , Metástasis Linfática , Estadificación de Neoplasias , Reacción en Cadena de la Polimerasa , Pronóstico , Estudios RetrospectivosRESUMEN
The rat olfactory bulb is an area displaying a particularly high density of substance P receptors in the glomerular cell layer whose functions are unknown. In pilot in vivo experiments we discovered that iontophoretically administered substance P potently depressed the spontaneous firing rate of most unidentified neurons of the rat olfactory bulb. To further elucidate the mechanism of this unexpected depressant effect, we studied the peptide's action in vitro on coronal sections of this brain region. Bath applied and microiontophoretically administered substance P depressed the spontaneous discharge of unidentified glomerular neurons in a dose-dependent fashion. This inhibiting effect is mediated indirectly via the release of another transmitter because it was abolished completely if the standard perfusion medium was replaced by a medium containing zero calcium and high magnesium. It appears that substance P acts by means of releasing GABA which in turn evokes the observed cell depression because the depressant effects were completely abolished by bath-applied bicuculline (10 microM) and picrotoxin (100 microM). In conclusion we propose that substance P indirectly depresses neuronal activity in the glomerular cell layer of the rat olfactory bulb by releasing gamma-aminobutyric acid.
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Bulbo Olfatorio/efectos de los fármacos , Sustancia P/farmacología , Ácido gamma-Aminobutírico/metabolismo , Potenciales de Acción/efectos de los fármacos , Animales , Bicuculina/farmacología , Técnicas In Vitro , Iontoforesis , Masculino , Neuronas/efectos de los fármacos , Neuronas/fisiología , Bulbo Olfatorio/fisiología , Ratas , Sustancia P/fisiologíaRESUMEN
The sensitivity of cingulate cortical neurons to microiontophoretically administered noradrenaline (NA) was compared in groups of rats treated with one or other of the two antidepressants mianserin (30 mg/kg s.c.) or iprindole (10 mg/kg i.p.) either acutely or chronically, for 10 and 28 days respectively. In the rats treated chronically with mianserin, the neurons were significantly less sensitive to NA than those of the acutely treated animals. By contrast, chronic administration of iprindole did not significantly affect neuronal sensitivity to NA.
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Encéfalo/efectos de los fármacos , Dibenzazepinas/farmacología , Indoles/farmacología , Iprindol/farmacología , Mianserina/farmacología , Neuronas/efectos de los fármacos , Norepinefrina/fisiología , Animales , Masculino , Ratas , Factores de TiempoRESUMEN
Racemic baclofen, (-)-baclofen and muscimol depressed all spontaneously firing locus coeruleus neurons tested in a slice preparation. Racemic phaclofen (100 microM; 1 mM) moderately antagonized the effects of racemic baclofen without antagonizing those of muscimol. Bicuculline (10, 30, 100 microM) potently antagonized the action of muscimol without affecting the inhibition of baclofen. Phaclofen and bicuculline had no pronounced effect on the spontaneous discharge rate of cells. The results suggests that there are GABAA and GABAB receptors in the locus coeruleus.
Asunto(s)
Locus Coeruleus/metabolismo , Receptores de GABA-A/metabolismo , Animales , Baclofeno/análogos & derivados , Baclofeno/farmacología , Técnicas In Vitro , Locus Coeruleus/efectos de los fármacos , Masculino , Muscimol/farmacología , Neuronas/efectos de los fármacos , Ratas , Ratas EndogámicasRESUMEN
The biochemical, electrophysiological and pharmacological properties of the new GABAB receptor blocker CGP 35348 are described. In a variety of receptor binding assays CGP 35348 showed affinity for the GABAB receptor only. CGP 35348 had an IC50 of 34 microM at the GABAB receptor. The compound antagonized (100, 300, 1000 microM) the potentiating effect of L-baclofen on noradrenaline-induced stimulation of adenylate cyclase in rat cortex slices. In electrophysiological studies CGP 35348 (10, 100 microM) antagonized the effect of L-baclofen in the isolated rat spinal cord. In the hippocampal slice preparation CGP 35348 (10, 30, 100 microM) blocked the membrane hyperpolarization induced by D/L-baclofen (10 microM) and the late inhibitory postsynaptic potential. CGP 35348 appeared to be 10-30 times more potent than the GABAB receptor blocker phaclofen. Ionophoretic and behavioural experiments showed that GABAB receptors in the brain were blocked after i.p. administration of CGP 35348. This compound may be of considerable value in elucidating the roles of brain GABAB receptors.
Asunto(s)
Anticonvulsivantes/farmacología , Compuestos Organofosforados/farmacología , Receptores de GABA-A/efectos de los fármacos , Inhibidores de Adenilato Ciclasa , Animales , Animales Recién Nacidos/metabolismo , Baclofeno/análogos & derivados , Baclofeno/farmacología , Conducta Animal/efectos de los fármacos , Química Encefálica/efectos de los fármacos , AMP Cíclico/metabolismo , Electrofisiología , Hipocampo/efectos de los fármacos , Técnicas In Vitro , Iontoforesis , Equilibrio Postural/efectos de los fármacos , Ensayo de Unión Radioligante , Ratas , Ratas Endogámicas , Médula Espinal/efectos de los fármacos , Médula Espinal/metabolismoRESUMEN
The goal of this report is to present the results obtained with three new GABAB receptor antagonists. CGP 54062 has an IC50 in a GABAB binding test of 0.013 microM which is roughly 2500-fold lower than one of the most potent blockers known so far, CGP 35348 (IC50 = 34 microM). CGP 46381 and CGP 36742 have IC50s of 4.9 and 36 microM respectively. The latter two compounds are the first orally active GABAB receptor antagonists. All three compounds bind to the GABAB receptor selectively, and are inactive in a number of binding tests assessing the compounds' affinity to various other receptor sites. The effect of these blockers on GABAergic transmission was investigated in the CA1 area of hippocampal slices. The Schaffer collateral/commissural fibers were stimulated and the evoked postsynaptic potentials were recorded intracellularly in pyramidal neurons. The three antagonists blocked the late inhibitory postsynaptic potential with the following rank order of potency CGP 54062 > 46381 > 36742 approximately 35348. These findings support the hypothesis that these potentials are mediated by GABAB receptors. Orally administered CGP 36742 and CGP 46381 block the neuronal depression induced by iontophoretically applied baclofen in anaesthetised rats. Up to a dose of 10 mg/kg i.v. CGP 54062 was inactive and thus does not appear to cross the blood-brain barrier at this dose. In anaesthetised rats the effects of the three new GABAB antagonists and of CGP 35348 were investigated on the paired-pulse inhibition of the population spikes evoked in the CA1 area of the hippocampus.(ABSTRACT TRUNCATED AT 250 WORDS)