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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
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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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
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
BACKGROUND: The primary goals of this paper are to describe the collection and evaluation of various nonpharmacological treatment options for depressive disorders and to establish a basis for the development of a standard for the treatment of patients with depressive disorders. METHOD: To identify evidence-based treatment elements, a comprehensive investigation of national and international guidelines was conducted. The extracted guidelines were then assessed with regard to aspects of methodological quality and evidence-based treatment elements. In a further step, specific and systematic literature searches for residual treatment elements were conducted. For the corresponding literature search, a hierarchical approach was chosen in which current guidelines were reviewed first and systematic reviews and meta-analyses second. Psychopharmacological treatments were excluded from the analysis because this is covered by specific guidelines. RESULTS: The treatment elements with an adequate level of evidence were identified as follows: psychotherapeutic interventions, marital/couples/family therapy and counseling, inclusion of family members, psycho-education, exercise, problem solving therapy, guided self-help and behavioral activation treatments. Further evidence-based methods include diagnostic treatment elements, participative decision-making, development of the therapeutic alliance, Cognitive Behavioral Analysis System for Psychotherapy, computerized cognitive behavior therapy, psychopharmacological therapy, combined psychopharmacological and psychotherapeutic therapy, electroconvulsive therapy, phototherapy, sleep deprivation, repetitive trans-cranial magnetic stimulation (rTMS) and acupuncture. CONCLUSION: In summary, using a hierarchical approach, it was possible to assign different levels of evidence to the various treatment options for depression.
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Trastorno Depresivo/terapia , Medicina Basada en la Evidencia/métodos , Psicoterapia/métodos , HumanosRESUMEN
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
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
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
IB1/JIP-1 is a scaffold protein that interacts with upstream components of the c-Jun N-terminal kinase (JNK) signaling pathway. IB1 is expressed at high levels in pancreatic beta cells and may therefore exert a tight control on signaling events mediated by JNK in these cells. Activation of JNK by interleukin 1 (IL-1beta) or by the upstream JNK constitutive activator DeltaMEKK1 promoted apoptosis in two pancreatic beta cell lines and decreased IB1 content by 50-60%. To study the functional consequences of the reduced IB1 content in beta cell lines, we used an insulin-secreting cell line expressing an inducible IB1 antisense RNA that lead to a 38% IB1 decrease. Reducing IB1 levels in these cells increased phosphorylation of c-Jun and increased the apoptotic rate in presence of IL-1beta. Nitric oxide production was not stimulated by expression of the IB1 antisense RNA. Complementary experiments indicated that overexpression of IB1 in insulin-producing cells prevented JNK-mediated activation of the transcription factors c-Jun, ATF2, and Elk1 and decreased IL-1beta- and DeltaMEKK1-induced apoptosis. These data indicate that IB1 plays an anti-apoptotic function in insulin-producing cells probably by controlling the activity of the JNK signaling pathway.
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Apoptosis/fisiología , Insulina/metabolismo , Interleucina-1/farmacología , Islotes Pancreáticos/efectos de los fármacos , Proteínas Nucleares/fisiología , Transactivadores/fisiología , Apoptosis/efectos de los fármacos , Doxiciclina/farmacología , Glucosa/farmacología , Secreción de Insulina , Islotes Pancreáticos/citología , Islotes Pancreáticos/metabolismo , Islotes Pancreáticos/efectos de la radiación , Óxido Nítrico/metabolismo , Fosforilación , Proteínas Serina-Treonina Quinasas/metabolismo , ARN sin Sentido/genética , Rayos UltravioletaRESUMEN
IB1/JIP-1 is a scaffold protein that regulates the c-Jun NH(2)-terminal kinase (JNK) signaling pathway, which is activated by environmental stresses and/or by treatment with proinflammatory cytokines including IL-1beta and TNF-alpha. The JNKs play an essential role in many biological processes, including the maturation and differentiation of immune cells and the apoptosis of cell targets of the immune system. IB1 is expressed predominantly in brain and pancreatic beta-cells where it protects cells from proapoptotic programs. Recently, a mutation in the amino-terminus of IB1 was associated with diabetes. A novel isoform, IB2, was cloned and characterized. Overall, both IB1 and IB2 proteins share a very similar organization, with a JNK-binding domain, a Src homology 3 domain, a phosphotyrosine-interacting domain, and polyacidic and polyproline stretches located at similar positions. The IB2 gene (HGMW-approved symbol MAPK8IP2) maps to human chromosome 22q13 and contains 10 coding exons. Northern and RT-PCR analyses indicate that IB2 is expressed in brain and in pancreatic cells, including insulin-secreting cells. IB2 interacts with both JNK and the JNK-kinase MKK7. In addition, ectopic expression of the JNK-binding domain of IB2 decreases IL-1beta-induced pancreatic beta-cell death. These data establish IB2 as a novel scaffold protein that regulates the JNK signaling pathway in brain and pancreatic beta-cells and indicate that IB2 represents a novel candidate gene for diabetes.
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Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Cromosomas Humanos Par 22 , Proteínas Nucleares/genética , Transactivadores/genética , Apoptosis , Secuencia de Bases , Northern Blotting , Mapeo Cromosómico , Clonación Molecular , Humanos , Insulina/metabolismo , Secreción de Insulina , Interleucina-1/metabolismo , Islotes Pancreáticos/metabolismo , Proteínas Quinasas JNK Activadas por Mitógenos , Proteínas Quinasas Activadas por Mitógenos/metabolismo , Datos de Secuencia Molecular , Proteínas Nucleares/metabolismo , Especificidad de Órganos , Isoformas de Proteínas/genética , Isoformas de Proteínas/metabolismo , Empalme del ARN , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , Transactivadores/metabolismo , Células Tumorales CultivadasRESUMEN
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
PURPOSE: To describe a method of delivering nitric oxide during high frequency jet ventilation. CLINICAL FEATURES: A 63-yr-old man underwent reduction pneumoplasty for bullous emphysema. Postoperatively, ventilation was inadequate, secondary to bilateral high output bronchopleural fistulae. High frequency jet ventilation was initiated and achieved adequate ventilation (pH>7.2). Over the following 24 hr, progressive hypoxemia (SaO2 <86%) developed along with the acute respiratory distress syndrome. Nitric oxide was delivered by continuous flow at the patient Y-connector during combined high frequency jet and conventional ventilation (two conventional low volume breaths/minute). Substantial improvement in oxygenation (FiO2 0.8 0.5, SaO2 >92%) was noted initially and was sustained over 72 hr. Subsequently, the patient was weaned to conventional ventilation without difficulty. Mechanical ventilation was discontinued on postoperative day sixteen. CONCLUSION: The simultaneous use of nitric oxide and high-frequency jet ventilation was used safely and effectively in this patient as a method of support for acute respiratory distress syndrome with co-existing large bilateral bronchopleural fistulae.
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Fístula Bronquial/terapia , Ventilación con Chorro de Alta Frecuencia , Óxido Nítrico/administración & dosificación , Enfermedades Pleurales/terapia , Síndrome de Dificultad Respiratoria/terapia , Humanos , Masculino , Persona de Mediana EdadRESUMEN
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
Islet-brain 1 (IB1), a regulator of the pancreatic beta-cell function in the rat, is homologous to JIP-1, a murine inhibitor of c-Jun amino-terminal kinase (JNK). Whether IB1 and JIP-1 are present in humans was not known. We report the sequence of the 2133-bp human IB1 cDNA, the expression, structure, and fine-mapping of the human IB1 gene, and the characterization of an IB1 pseudogene. Human IB1 is 94% identical to rat IB1. The tissue-specific expression of IB1 in human is similar to that observed in rodent. The IB1 gene contains 12 exons and maps to chromosome 11 (11p11.2-p12), a region that is deleted in DEFECT-11 syndrome. Apart from an IB1 pseudogene on chromosome 17 (17q21), no additional IB1-related gene was found in the human genome. Our data indicate that the sequence and expression pattern of IB1 are highly conserved between rodent and human and provide the necessary tools to investigate whether IB1 is involved in human diseases.
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Proteínas Adaptadoras Transductoras de Señales , Proteínas Portadoras/genética , Proteínas Portadoras/metabolismo , Cromosomas Humanos Par 11 , Proteínas Nucleares/genética , Proteínas Nucleares/metabolismo , Transactivadores/genética , Transactivadores/metabolismo , Animales , Secuencia de Bases , Northern Blotting , Southern Blotting , Mapeo Cromosómico , Cromosomas Bacterianos/genética , ADN Complementario/análisis , Exones/genética , Humanos , Hibridación Fluorescente in Situ , Intrones/genética , Ratones , Datos de Secuencia Molecular , Seudogenes , Ratas , Análisis de Secuencia de ADN , Distribución TisularRESUMEN
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
Interactive behavior of 30 mothers of infants with mental delay and 30 comparison mothers and infants was examined in relation to child age (first and second year), context (feeding versus teaching), maternal characteristics (family stress, coping resources), and family social system (maternal education). Groups were compared from two perspectives: with infants matched on mental age (9 and 19 months MA), and on chronological age (8 and 18 months CA). Study mothers scored lower than comparison mothers during teaching but not during feeding in year 1 with both MA and CA match, but only with the CA match in year 2. Study infants scored lower than comparison infants in both contexts in year 1, but not in year 2. Groups did not differ on maternal or family measures. In year 1, group status, coping, and maternal education were predictive of mother interaction. In year 2, only maternal education was predictive. Results confirm the importance of type of match, context, and family system variables in understanding effects of child mental delay on maternal interactive behavior.
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Discapacidades del Desarrollo , Relaciones Madre-Hijo , Adaptación Psicológica , Adulto , Factores de Edad , Escolaridad , Conducta Alimentaria , Femenino , Humanos , Lactante , Masculino , Estrés PsicológicoRESUMEN
OBJECTIVE: Parasitic infestations are known to elicit T-helper lymphocyte type 2 (Th-2) reactions, characterized by a pronounced eosinophila and high IgE levels. In humans both elevated specific IgE levels and eosinophil counts are associated with resistance to reinfection with schistosomiasis. This study aimed to establish whether the Th-2 reaction could be enhanced with calcitriol (vitamin D3). Calcitriol has been shown to cause a shift from Th-1 to Th-2 type reactions when applied locally to the skin. METHODS: Fifty-nine patients with Schistostoma haematobium infection were randomized to one of four treatment modalities, i.e. (a) praziquantel (PZQ) 60 mg.kg-1 orally on day 1, (b) PZQ 60 mg.kg-1 on day 1 plus calcitriol 1 microgram per day orally for 5 consecutive days, (c) calcitriol 1 microgram per day for 5 consecutive days or (d) placebo. Blood for differential counts, eosinophil cationic protein (ECP), specific IgE and IgG to whole-worm antigen, as well as urine samples for egg counts, were collected on days 0 and 21. RESULTS: Baseline values did not differ significantly between the groups. Calcitriol alone resulted in significant increases in circulating lymphocytes (median increase of 5.5%) and the percentage of eosinophil vacuolization (mean increase 28%). It, however, significantly decreased ECP levels (mean decrease 46%). PZQ in combination with calcitriol significantly enhanced production of specific IgE (mean increase 213%) and IgG (mean increase of 170%) and tended to increase eosinophil vacuolization (mean increase 22%). All these changes also differed significantly from those in the placebo group. The specific IgE and IgG levels were also significantly higher than the already increased levels seen with PZQ treatment only. ECP levels were, however, not significantly affected by combination therapy, whereas PZQ alone significantly enhanced ECP production (mean increase 93%). CONCLUSIONS: The increases in specific IgE responses and percentage of eosinophil vacuolization favour a Th-2 type of reaction. The ECP values viewed in isolation may, paradoxically, indicate a Th-1 response; this could, however, have been an artefact due to the method of ECP detection ex vivo. Finally, it would seem that calcitriol does cause some immune augmentation when combined with PZQ therapy in patients with schistosomiasis. However, long-term follow-up is needed to prove that these findings would translate into resistance against re-infection.
Asunto(s)
Calcitriol/farmacología , Eosinófilos/efectos de los fármacos , Inmunoglobulina E/metabolismo , Ribonucleasas , Esquistosomiasis Urinaria/inmunología , Adolescente , Proteínas Sanguíneas/metabolismo , Calcitriol/administración & dosificación , Calcitriol/uso terapéutico , Ensayo de Inmunoadsorción Enzimática , Proteínas en los Gránulos del Eosinófilo , Eosinófilos/metabolismo , Humanos , Inmunoglobulina G/metabolismo , Recuento de Leucocitos , Recuento de Linfocitos , Masculino , Recuento de Huevos de Parásitos , Praziquantel/uso terapéutico , Esquistosomiasis Urinaria/sangre , Esquistosomiasis Urinaria/orina , Esquistosomicidas/administración & dosificación , Esquistosomicidas/uso terapéutico , Método Simple CiegoRESUMEN
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.