Asunto(s)
Pénfigo , Humanos , Desmocolinas , Desmogleína 1 , Desmogleína 3 , Anticuerpos , AutoanticuerposRESUMEN
AIMS: The purpose of this study was to clarify the clinical behaviour, prognosis, and optimum treatment of dedifferentiated low-grade osteosarcoma (DLOS) diagnosed based on molecular pathology. PATIENTS AND METHODS: We retrospectively reviewed 13 DLOS patients (six men, seven women; median age 32 years (interquartile range (IQR) 27 to 38)) diagnosed using the following criteria: the histological coexistence of low-grade and high-grade osteosarcoma components in the lesion, and positive immunohistochemistry of mouse double minute 2 homolog (MDM2) and cyclin-dependent kinase 4 (CDK4) associated with MDM2 amplification. These patients were then compared with 51 age-matched consecutive conventional osteosarcoma (COS) patients (33 men, 18 women; median age 25 years (IQR 20 to 38)) regarding their clinicopathological features. RESULTS: The five-year overall survival (OAS) rates in the DLOS and COS patients were 85.7% and 77.1% (p = 0.728), respectively, and the five-year progression-free survival (PFS) rates were 57.7% and 44.9% (p = 0.368), respectively. A total of 12 DLOS patients received chemotherapy largely according to regimens for COS. Among the nine cases with a histological evaluation after chemotherapy, eight showed a poor response, and seven of these had a necrosis rate of < 50%. One DLOS patient developed local recurrence and five developed distant metastases. CONCLUSION: Based on our study of 13 DLOS cases that were strictly defined by histological and molecular means, DLOS showed a poorer response to a standard chemotherapy regimen than COS, while the clinical outcomes were not markedly different. Cite this article: Bone Joint J 2019;101-B:745-752.
Asunto(s)
Neoplasias Óseas/patología , Neoplasias Óseas/terapia , Osteosarcoma/patología , Osteosarcoma/terapia , Adulto , Biomarcadores de Tumor/análisis , Estudios de Casos y Controles , Terapia Combinada , Femenino , Humanos , Inmunohistoquímica , Masculino , Clasificación del Tumor , Estudios Retrospectivos , Tasa de SupervivenciaRESUMEN
It is well known that early life events induce long-lasting psychophysiological and psychobiological influences in later life. In rodent studies, environmental enrichment after weaning prevents the adulthood behavioral and emotional disturbances in response to early adversities. We compared the behavioral effect of neonatal isolation (NI) with the effect of NI accompanied by tactile stimulation (NTS) to determine whether NTS could reverse or prevent the effects of NI on the adulthood behavioral and emotional responses to environmental stimuli. In addition, we also examined the sex difference of the NTS effect. Measurements of body weights, an open-field locomotor test, an elevated plus maze test, a hot-plate test, and a contextual fear-conditioning test were performed on postnatal day 60. As compared with rats subjected to NI, rats subjected to NTS showed significantly higher activity and exploration in the open-field locomotor test, lower anxiety-like behavior in the elevated plus maze test, and significantly prolonged latencies in the hot-plate test, and this effect was equal among males and females. In the contextual fear-conditioning test, whereas NTS significantly reduced the enhanced freezing time due to NI in females, no significant difference in the freezing time between NI and NTS was found in males. These findings indicate that adequate tactile stimulation in early life plays an important role in the prevention of disturbances in the behavioral and emotional responses to environmental stimuli in adulthood induced by early adverse experiences.
Asunto(s)
Animales Recién Nacidos/fisiología , Ansiedad/psicología , Umbral del Dolor/psicología , Aislamiento Social/psicología , Tacto/fisiología , Análisis de Varianza , Animales , Animales Recién Nacidos/psicología , Ansiedad/etiología , Condicionamiento Clásico/fisiología , Ambiente , Conducta Exploratoria/fisiología , Femenino , Masculino , Umbral del Dolor/fisiología , Ratas , Ratas Sprague-Dawley , Factores Sexuales , Estadísticas no ParamétricasRESUMEN
Neural stem cell (NSC) transplantation has been investigated and developed in areas such as brain injury, stroke and neurodegenerative diseases. Recently, emerging evidence suggest that many of clinical symptoms observed in psychiatric disease are likely related to neural network disruptions including neurogenesis dysfunction. In the present study, we transplanted NSCs into a model of fetal alchol effects (FAE) for the purpose of investigating the possibility of regenerative therapy for the FAE. We labeled NSCs with fluorescent dye and radioisotope which were transplanted into FAE rats by intravenous injection. The transplanted cells were detected in wide areas of brain and were greater in number in the brains of the FAE group compared to the control group. Furthermore NSC transplantation attenuated behavioral abnormalities in FAE animals. These results suggest NSC transplantation as a potental new therapy for human FAE.
Asunto(s)
Modelos Animales de Enfermedad , Células Madre Embrionarias/trasplante , Trastornos del Espectro Alcohólico Fetal/cirugía , Regeneración Nerviosa/fisiología , Neuronas/citología , Animales , Animales Recién Nacidos , Encéfalo/patología , Diferenciación Celular/fisiología , Movimiento Celular/fisiología , Células Madre Embrionarias/patología , Conducta Exploratoria/fisiología , Miedo/fisiología , Femenino , Trastornos del Espectro Alcohólico Fetal/patología , Humanos , Inyecciones Intravenosas , Aprendizaje por Laberinto/fisiología , Microscopía Fluorescente , Neuronas Motoras/fisiología , Neuronas/patología , Embarazo , Ratas , Ratas WistarRESUMEN
Recent clinical neuroimaging studies have suggested that morphological brain changes occur and progress in the course of schizophrenia. Although the neurogenetic and neurotrophic effects of antipsychotics are considered to contribute to the prevention of reduction in brain volume, the cellular molecular mechanisms of action of antipsychotics have not yet been elucidated. We examined the effects of antipsychotics on the endoplasmic reticulum (ER) stress-induced damages of neurons and neural stem cells (NSCs) using cultured cells. In the neuronal cultures, the atypical antipsychotic olanzapine protected neurons from thapsigargin (1 microM)-induced injury. It was observed that a low concentration of thapsigargin (10 nM) that did not affect the neuronal survival could reduce neuronal differentiation of cultured NSCs, suggesting a role of ER stress in the differentiation function of NSCs. Treatment with olanzapine increased the neuronal differentiation suppressed by the exposure to thapsigargin (10 nM). The thapsigargin-induced ER chaperones, GRP78, which indicate the ER stress condition of the cell, were decreased by the treatment with the atypical antipsychotics olanzapine and quetiapine but not by the typical antipsychotic haloperidol. These results indicate that the amelioration of ER-stress might be involved in the cellular mechanisms of atypical antipsychotics to produce neuroprotective and neurogenetic actions in neurons and NSCs, suggesting potential roles of these drugs for treatment of schizophrenia.
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Antipsicóticos/farmacología , Benzodiazepinas/farmacología , Diferenciación Celular/efectos de los fármacos , Supervivencia Celular/efectos de los fármacos , Retículo Endoplásmico/efectos de los fármacos , Proteínas de Choque Térmico/biosíntesis , Neuronas/efectos de los fármacos , Células Madre/efectos de los fármacos , Animales , Diferenciación Celular/fisiología , Supervivencia Celular/fisiología , Células Cultivadas , Sinergismo Farmacológico , Retículo Endoplásmico/fisiología , Femenino , Proteínas de Choque Térmico/fisiología , Neuronas/fisiología , Olanzapina , Embarazo , Ratas , Ratas Wistar , Células Madre/citología , Células Madre/fisiologíaRESUMEN
A 37-year-old man was admitted to our hospital. The patient noted sudden right back pain after coughing before 1 hour. Loss of consciousness was occurred in an ambulance. Chest X-P revealed whole fluid in the right chest. Enhanced chest computed tomography (CT) revealed extravasation of contrast media into the pleural cavity from the right chest wall. Thoracentesis was performed to relieve dyspnea and 2,000 ml of blood was removed. Then hemoglobin count was dropped to 3.8 g/dl. At thoracotomy whole blood was sucked about 3,900 ml. Bleeding point was found at third intercostal vein. The vein was knotted and sutured by prolene thread. The bleeding lesion was no inflammation and no string like tissue. We report a case of idiopathic hemothorax and enhanced chest CT was useful for diagnosis of bleeding lesion of pleural cavity.
Asunto(s)
Hemotórax/diagnóstico por imagen , Choque/etiología , Tomografía Computarizada por Rayos X , Adulto , Ambulancias , Hemotórax/cirugía , Humanos , Masculino , Intensificación de Imagen RadiográficaRESUMEN
The effects of microbial transglutaminase (MTGase) on type I collagen self-assembly and properties of reconstructed collagen fibrils from shark were investigated. Collagen self-assembly was accelerated with the addition of MTGase in dependence on that concentration. The relative amount of reconstructed collagen slightly decreased with MTGase. The diffusion coefficient of collagen gel was reduced by treatment with MTGase, and that suggested the reduction of mobility of the whole collagen network. At a high temperature, used to denature the collagen, MTGase-treated collagen gel remained as aggregates. By differential scanning calorimetry, the denaturation temperature of MTGase-treated collagen gel was about 2 degrees C higher than that of nontreated collagen gel. Treatment with MTGase yielded thermally stable cross-links in collagen molecules.
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Colágeno Tipo I/química , Colágeno Tipo I/metabolismo , Tiburones , Transglutaminasas/metabolismo , Animales , Rastreo Diferencial de Calorimetría , Colágeno Tipo I/ultraestructura , Hongos/enzimología , Geles/química , Geles/metabolismo , Microscopía Electrónica de Rastreo , Reología , Dispersión de Radiación , Temperatura , Resistencia a la Tracción , Factores de TiempoRESUMEN
The C(4)-Pdk gene encoding the C(4) enzyme pyruvate, orthophosphate dikinase (PPDK) of maize (Zea mays cv Golden Cross Bantam) was introduced into the C(3) plant, rice (Oryza sativa cv Kitaake). When the intact maize C(4)-Pdk gene, containing its own promoter and terminator sequences and exon/intron structure, was introduced, the PPDK activity in the leaves of some transgenic lines was greatly increased, in one line reaching 40-fold over that of wild-type plants. In a homozygous line, the PPDK protein accounted for 35% of total leaf-soluble protein or 16% of total leaf nitrogen. In contrast, introduction of a chimeric gene containing the full-length cDNA of the maize PPDK fused to the maize C(4)-Pdk promoter or the rice Cab promoter only increased PPDK activity and protein level slightly. These observations suggest that the intron(s) or the terminator sequence of the maize gene, or a combination of both, is necessary for high-level expression. In maize and transgenic rice plants carrying the intact maize gene, the level of transcript in the leaves per copy of the maize C(4)-Pdk gene was comparable, and the maize gene was expressed in a similar organ-specific manner. These results suggest that the maize C(4)-Pdk gene behaves in a quantitatively and qualitatively similar way in maize and transgenic rice plants. The activity of the maize PPDK protein expressed in rice leaves was light/dark regulated as it is in maize. This is the first reported evidence for the presence of an endogenous PPDK regulatory protein in a C(3) plant.
Asunto(s)
Oryza/fisiología , Piruvato Ortofosfato Diquinasa/metabolismo , Zea mays/fisiología , Clorofila/metabolismo , Oscuridad , Regulación de la Expresión Génica de las Plantas , Intrones , Luz , Complejos de Proteína Captadores de Luz , Nitrógeno/metabolismo , Oryza/genética , Fotosíntesis/fisiología , Proteínas del Complejo del Centro de Reacción Fotosintética , Hojas de la Planta/genética , Hojas de la Planta/fisiología , Proteínas de Plantas/aislamiento & purificación , Proteínas de Plantas/metabolismo , Plantas Modificadas Genéticamente , Ribulosa-Bifosfato Carboxilasa/metabolismo , Zea mays/genéticaRESUMEN
CalDAG-GEFI and CalDAG-GEFII (identical to RasGRP) are novel, brain-enriched guanine nucleotide exchange factors (GEFs) that can be stimulated by calcium and diacylglycerol and that can activate small GTPases, including Ras and Rap1, molecules increasingly recognized as having signaling functions in neurons. Here, we show that CalDAG-GEFI and CalDAG-GEFII mRNAs, detected by in situ hybridization analysis, have sharply contrasting forebrain-predominant distributions in the mature brain: CalDAG-GEFI is expressed mainly in the striatum and olfactory structures and deep cortical layers, whereas CalDAG-GEFII is expressed widely in the forebrain. Within the striatum, however, the two CalDAG-GEF mRNAs have nearly identical distributions: they are coexpressed in striatal projection neurons that give rise to the direct and indirect pathways of the basal ganglia. Subcellular fractionation analysis of the substantia nigra with monoclonal antibodies against CalDAG-GEFI suggests that CalDAG-GEFI protein is present not only in the cell bodies of striatal projection neurons but also in their axons and axon terminals. These results suggest that the CalDAG-GEFs may be key intracellular regulators whereby calcium and diacylglycerol signals can regulate cellular functions through small GTPases in the basal ganglia circuits.
Asunto(s)
Cuerpo Estriado/citología , Factores de Intercambio de Guanina Nucleótido/análisis , Proteínas del Tejido Nervioso/análisis , Neuronas/química , Factores de Edad , Animales , Ganglios Basales/metabolismo , Western Blotting , Cuerpo Estriado/embriología , Cuerpo Estriado/crecimiento & desarrollo , Cuerpo Estriado/metabolismo , Vías Eferentes/química , Electroforesis en Gel de Poliacrilamida , Regulación del Desarrollo de la Expresión Génica , Factores de Intercambio de Guanina Nucleótido/biosíntesis , Factores de Intercambio de Guanina Nucleótido/genética , Hibridación in Situ , Proteínas del Tejido Nervioso/biosíntesis , Proteínas del Tejido Nervioso/genética , Ratas , Ratas Sprague-Dawley , Fracciones Subcelulares/química , Proteínas de Unión al GTP rap1/fisiología , Factores de Intercambio de Guanina Nucleótido ras , Proteínas ras/fisiologíaRESUMEN
RP-1776, a novel cyclic peptide, was isolated from the culture broth of Streptomyces sp. KY11784. RP-1776 selectively inhibited the binding of PDGF BB to the extracellular domain of the PDGF beta-receptor with an IC50 value of 11 +/- 6 microM. Detailed binding experiments suggested that RP-1776 directly interacts with PDGF BB. RP-1776 inhibited the phosphorylation of the PDGF beta-receptor induced by PDGF BB. These results suggested that RP-1776 antagonizes the signaling of PDGF BB probably through the inhibition of PDGF BB binding to the PDGF beta-receptor.
Asunto(s)
Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Péptidos Cíclicos/aislamiento & purificación , Péptidos Cíclicos/farmacología , Factor de Crecimiento Derivado de Plaquetas/antagonistas & inhibidores , Receptores del Factor de Crecimiento Derivado de Plaquetas/metabolismo , Streptomyces/química , Animales , Antibacterianos/química , Becaplermina , Células CHO , Cricetinae , Depsipéptidos , Fermentación , Concentración 50 Inhibidora , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Péptidos Cíclicos/química , Fosforilación/efectos de los fármacos , Factor de Crecimiento Derivado de Plaquetas/metabolismo , Estructura Terciaria de Proteína , Proteínas Proto-Oncogénicas c-sis , Microbiología del Suelo , Streptomyces/metabolismoRESUMEN
In this study, we attempted to determine the prevalence of tobacco or nicotine dependence in current smokers in Japan and to assess the relationship between alcoholism and tobacco or nicotine dependence. The subjects consisted of 246 alcohol-dependent and 1,111 non-alcohol-dependent individuals. We used a questionnaire, consisting of items obtained from the World Health Organization's The ICD-10 Classification of Mental and Behavioural Disorders: Clinical Descriptions and Diagnostic Guidelines (ICD-10) and the American Psychiatric Association's Diagnostic and Statistical Manual of Mental Disorders (4th ed.; DSM-IV) criteria for tobacco or nicotine dependence. The prevalence of tobacco dependence diagnosed according to the ICD-10 criteria was 23.9% among all subjects. The prevalence of tobacco dependence diagnosed according to the ICD-10 criteria was higher in alcohol-dependent individuals (58.1%) than in nondrinkers or social drinkers (12.8%). Alcohol-dependent subjects consumed significantly more nicotine per day than did nondrinkers or social drinkers. The prevalence of nicotine physical dependence diagnosed by using DSM-IV criteria for nicotine withdrawal was 2.4% in alcohol-dependent individuals, whereas only 0.3% of nondrinkers or social drinkers exhibited nicotine physical dependence. These results indicate to us that the potential for nicotine physical dependence is not much stronger than that reported among current smokers.
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Fumar/epidemiología , Tabaquismo/epidemiología , Adulto , Anciano , Humanos , Japón , Persona de Mediana Edad , Nicotina/efectos adversos , Prevalencia , Cese del Hábito de Fumar/estadística & datos numéricos , Síndrome de Abstinencia a Sustancias/epidemiología , Tabaquismo/diagnósticoRESUMEN
Western blot analysis was performed by using a specific antibody to measure annexin IV in human postmortem brain samples from alcoholic subjects. The analysis showed a significantly augmented expression in the hippocampus compared with controls, whereas the expression in the frontal cortex was equivalent in both groups. Annexin IV expression in the occipital cortex tended to increase in alcoholics. It was shown further that autoantibodies to annexin IV were increased significantly in alcoholic patients compared with controls. Thus, annexin IV may become a novel biological marker for alcoholics.
Asunto(s)
Alcoholismo/metabolismo , Anexina A4/análisis , Química Encefálica , Adulto , Anciano , Alcoholismo/inmunología , Anexina A4/inmunología , Autoanticuerpos/sangre , Western Blotting , Membrana Celular/química , Lóbulo Frontal/química , Hipocampo/química , Humanos , Inmunohistoquímica , Persona de Mediana Edad , Lóbulo Occipital/químicaRESUMEN
In the presence of urea, type I collagen could form a gel with crosslinks with microbial transglutaminase (MTGase). Collagen self-assembly was accelerated with the addition of MTGase. The proportion of reconstructed collagen fibrils was raised with the addition of MTGase. MTGase-treated collagen gel remained gelled at high temperatures at which collagen denatured. By treatment with MTGase, collagen could form the gel under impossible condition to collagen self-assembly, and that denaturation temperature was raised.
Asunto(s)
Colágeno/química , Tiburones/metabolismo , Transglutaminasas/química , Urea/química , Animales , Elasticidad , Streptomyces/enzimología , ViscosidadRESUMEN
Using RACE techniques we have cloned and sequenced one of the hamster liver 3-hydroxy-hexobarbital dehydrogenases which catalyze not only cyclic alcohols but also 17beta-hydroxy-steroids and 3alpha-hydroxysteroids. The gene specific primers to 3-hydroxyhexobarbital dehydrogenase 1 (G2) were synthesized on the basis of its partial peptide sequences. The sequence of full length cDNA generated by 3'- and 5'-RACE PCR consisted of 1225 nucleotides including an open reading frame of 972 nucleotides encoding a protein of 323 amino acids. The deduced amino acid sequence matched exactly with the partial peptide sequences of hamster liver 3-hydroxyhexobarbital dehydrogenase 1 (G2). The sequence showed 84.5% identity to mouse liver 17beta-dehydrogenase(A-specific), and 74-76% identity to human liver bile acid binding protein/3alpha-hydroxysteroid dehydrogenase (DD2), human liver 3alpha-hydroxysteroid dehydrogenase type I (DD4) and type II (DD3), and rabbit ovary 20alpha-hydroxysteroid dehydrogenase. The protein contains catalytic residues of aldo-keto reductases, Asp50, Tyr55, Lys84, His117. These results suggest that the hamster liver 3-hydroxyhexobarbital/17beta(3alpha)-hydroxysteroid dehydrogenase belongs to aldo-keto reductase superfamily. The insert containing the full-length cDNA of 3-hydroxyhexobarbital dehydrogenase and vector specific overhang produced by PCR was annealed with pET-32 Xa/LIC vector. The plasmid was transformed into BL21 (DE3) cells containing pLysS. The recombinant enzyme was induced 1 mM IPTG. The expressed enzyme was produced as fusion protein and purified by nickel chelating affinity chromatography followed by POROS CM column chromatography and superdex 75 gel filtration. Molecular weight of the recombinant enzyme fused thioredoxin and his*tag was about 55000 and that was 35000 after Factor Xa protease treatment. The recombinant enzyme dehydrogenated 3-hydroxy-hexobarbital, 1-acenaphthenol, 2-cyclohexen-1-ol, testosterone, glycolithocholic acid as well as the native enzyme purified from hamster liver.
Asunto(s)
17-Hidroxiesteroide Deshidrogenasas/genética , 3-Hidroxiesteroide Deshidrogenasas/genética , Oxidorreductasas de Alcohol/genética , Hígado/enzimología , 17-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-Hidroxiesteroide Deshidrogenasas/metabolismo , 3-alfa-Hidroxiesteroide Deshidrogenasa (B-Específica) , Oxidorreductasas de Alcohol/metabolismo , Secuencia de Aminoácidos , Animales , Clonación Molecular , Cricetinae , ADN Complementario/genética , Escherichia coli/genética , Expresión Génica , Técnicas In Vitro , Datos de Secuencia Molecular , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Especificidad por SustratoRESUMEN
The chloroplastic NADP-malic enzyme (NADP-ME) is a key enzyme of the C4 photosynthesis pathway in NADP-ME type C4 plants such as maize. To express the chloroplastic NADP-ME in leaves of a C3 plant, rice, full-length cDNAs encoding the rice C3-specific isoform and the maize C4-specific isoform of the enzyme were expressed under the control of the rice CAB: promoter. Transformants carrying the rice cDNA showed the NADP-ME activities in the leaves less than several-fold that of non-transformants, while those carrying the maize cDNA showed activities up to 30-fold that of non-transformants or about 60% of the NADP-ME activity of maize leaves. These results indicate that expression of the rice C3-specific NADP-ME is suppressed at co- and/or post-transcriptional levels by some regulation mechanisms intrinsic to rice, while that of the foreign C4-specific isoform can escape from such suppression. The accumulation of the maize C4-specific NADP-ME led to bleaching of leaf color and growth hindrance in rice plants under natural light. These deteriorative effects resulted from enhanced photoinhibition of photosynthesis due to an increase in the level of NADPH inside the chloroplast by the action of the maize enzyme.
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Malato Deshidrogenasa/metabolismo , Oryza/enzimología , Expresión Génica , Malato Deshidrogenasa/genética , Oryza/crecimiento & desarrollo , Hojas de la Planta/enzimología , Plantas Modificadas GenéticamenteRESUMEN
Advanced glycation end products (AGE) and the receptor for AGE (RAGE) have been implicated in the chronic complications of diabetes mellitus (DM), and have been reported to play an important role in the pathogenesis of Alzheimer's disease (AD). In this study, we established a polyclonal anti-RAGE antibody, and examined the immunohistochemical localization of amyloid beta protein (Abeta), AGE, and RAGE in neurons and astrocytes from patients with AD and DM. Our anti-RAGE antibody recognized full-length RAGE (50 kd) and N-terminal RAGE (35 kd) in human brain tissue. Abeta-, AGE-, and RAGE-positive granules were identified in the perikaryon of hippocampal neurons (especially from CA3 and CA4) in all subjects. The distribution and staining pattern of these immunopositive granules showed good concordance with each antibody. In AD, most astrocytes contained both AGE-and RAGE-positive granules and their distribution was almost the same. Abeta-positive granules were less common, but Abeta-, AGE-, and RAGE-positive granules were colocalized in one part of a single astrocyte. In DM patients and control cases, AGE-and RAGE-positive astrocytes were very rare. These finding support the hypothesis that glycated Abeta is taken up via RAGE and is degraded through the lysosomal pathway in astrocytes. In addition to the presence of AGE, the process of AGE degradation and receptor-mediated reactions may contribute to neuronal dysfunction and promote the progression of AD.
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Enfermedad de Alzheimer/metabolismo , Astrocitos/metabolismo , Diabetes Mellitus/metabolismo , Neuronas/metabolismo , Receptores Inmunológicos/metabolismo , Anciano , Enfermedad de Alzheimer/patología , Péptidos beta-Amiloides/metabolismo , Animales , Especificidad de Anticuerpos , Astrocitos/patología , Western Blotting , Bovinos , Diabetes Mellitus/patología , Femenino , Productos Finales de Glicación Avanzada/metabolismo , Humanos , Inmunohistoquímica , Masculino , Persona de Mediana Edad , Neuronas/patología , Receptor para Productos Finales de Glicación AvanzadaRESUMEN
We investigated feeding-associated activation of neurons in the medulla oblongata during the suckling period in rats, using the c-fos gene-encoded protein (Fos) immunohistochemistry. After an isolation from mothers for 12 h, neonates were either breast-fed intensively or further isolated for another 3 h, and sacrificed on postnatal day 1, 3, 5, 7 and 14 (P1-14). In the former pups, Fos-immunoreactive (FI) neurons were predominantly localized in the nucleus of the solitary tract (NST) and in the dorsal medullary reticular formation (RF). The number of FI cells peaked on P5-7 and decreased on P14 in the NST, and increased remarkably on P3 and was consistently high until P14 in the dorsal RF. In contrast, much fewer FI cells were found in the NST and RF in the latter pups. The results indicated that not only the NST but also the dorsal RF were implicated in feeding behavior in the suckled pups.
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Ingestión de Alimentos/fisiología , Proteínas Proto-Oncogénicas c-fos/biosíntesis , Formación Reticular/metabolismo , Núcleo Solitario/metabolismo , Envejecimiento/fisiología , Animales , Animales Recién Nacidos , Animales Lactantes , Recuento de Células , Conducta Alimentaria/fisiología , Neuronas/citología , Neuronas/metabolismo , Ratas , Ratas Sprague-Dawley , Formación Reticular/citología , Núcleo Solitario/citologíaRESUMEN
The physicochemical properties of shark type I collagen gel and membrane were not same as those of pig type I collagen. The denaturation temperature of shark collagen gel was about 15 degrees C lower. According to scanning electronic micrography, the diameter of shark collagen fibril was relatively thin and more homogeneous. The breaking strength of shark collagen gel was greater, and shark collagen membrane had a greater mechanical strength and a higher water vapor sorption.
Asunto(s)
Colágeno/química , Animales , Rastreo Diferencial de Calorimetría , Colágeno/ultraestructura , Geles , Membranas Artificiales , Desnaturalización Proteica , Tiburones , Piel/químicaRESUMEN
Morphine 6-dehydrogenase, which catalyzes the dehydrogenation of morphine to morphinone, was purified 815-fold to a homogeneous protein from the soluble fraction of hamster liver with a yield of 15%. The enzyme was a monomeric protein with a molecular weight of 38 kDa and an isoelectric point of 5.6. Although both NAD and NADP served as cofactors, the enzyme activity with NADP was less than 5% that found with NAD at pH 7.4. With NAD, the enzyme gave the maximal activity at pH 9.3, and the K(m) and V(max) values toward morphine were 1.0 mM and 0.43 unit/mg protein, respectively. Among morphine congeners, normorphine exhibited higher activity than morphine, but codeine and ethylmorphine were poor substrates, and dihydromorphine and dihydrocodeine showed no detectable activity. The enzyme also exhibited significant activity for a variety of cyclic and alicyclic alcohols. In addition to xenobiotics, the enzyme catalyzed the dehydrogenation of 17beta-hydroxysteroids with much higher affinities than morphine. In the reverse reaction, the enzyme exhibited high activity for o-quinones, but morphinone, naloxone, and aromatic aldehydes and ketones were reduced at slow rates. Sulfhydryl reagents and ketamine strongly inhibited the enzyme, whereas pyrazole, barbital, and indomethacin had little effect on enzyme activity. 17beta-Hydroxysteroids inhibited the enzyme in a competitive manner against morphine. A total of 302 amino acid residues, which comprised approximately 94% of whole protein, were identified by sequencing of the peptides obtained by proteolytic digestion. This amino acid sequence of the enzyme showed significant homology to members of the aldo-keto reductase (AKR) superfamily and shared 63-64% identity with members of the AKR1C subfamily. These findings indicate that the enzyme is a new member of the AKR superfamily that is involved in steroid metabolism as 17beta-hydroxysteroid dehydrogenase as well as xenobiotic metabolism.
Asunto(s)
Oxidorreductasas de Alcohol/aislamiento & purificación , Oxidorreductasas de Alcohol/metabolismo , Hígado/enzimología , NAD/metabolismo , Oxidorreductasas de Alcohol/química , Secuencia de Aminoácidos , Animales , Cromatografía en Gel , Cromatografía por Intercambio Iónico , Cricetinae , Citosol/enzimología , Electroforesis en Gel de Poliacrilamida , Hidroxiesteroides/metabolismo , Masculino , Mesocricetus , Ratones , Datos de Secuencia Molecular , Peso Molecular , Morfina/metabolismo , Derivados de la Morfina/metabolismo , NADP/metabolismo , Ratas , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Especificidad por SustratoRESUMEN
Fibril reconstruction process, that is, the nucleation and growth of mixed type I collagen fibril of shark and pig, progressed faster than that of the individual collagen species of shark or pig. The reconstructed mixed collagen fibril had a greater resistance to return to the solution or to melt into gelatin in comparison with the counterpart consisting solely of shark collagen. The denaturation temperature of the mixed collagen gel was about 10 degrees C higher than that of shark, and about 5 degrees C lower than that of pig. By scanning electron microscopy, the diameter of mixed collagen fibril showed an intermediate range between shark and pig collagen fibril. The breaking strength of the mixed collagen gel was tougher than that of pig, but weaker than that of shark. Other physicochemical properties of the mixed type I collagen gel were observed to be at intermediate positions between those of shark and pig type I collagen gels.