RESUMEN
Rikkunshito has been shown to improve upper gastrointestinal symptoms and anorexia. The aim of this study was to evaluate whether rikkunshito improves chemotherapy-induced nausea in thoracic cancer patients receiving carboplatin (CBDCA)-based chemotherapy. A retrospective before-and-after comparison study was conducted in patients with thoracic cancer receiving the first cycle of CBDCA-based chemotherapy. Among 61 eligible patients, 34 received standard antiemetic therapy with a combination of 5-hydroxytryptamine-3 receptor antagonist and dexamethasone from September 2012 and June 2013 (standard group), while the other 27 received the standard antiemetic therapy plus oral rikkunshito from July 2013 and December 2014 (rikkunshito group). The rates of no nausea showed no significant difference between the standard and rikkunshito group (Overall phase: 64.7 % for standard group vs 74.1 % for rikkunshito group, p = 0.579). Subgroup analysis indicated that, in female patients, the rates of no nausea in rikkunshito groups was significantly higher than in standard group (overall phase: 44.4 % vs 100 %, p = 0.034). Rikkunshito did not demonstrate an additional prophylactic effect on standard antiemetic therapy for nausea in patients with thoracic cancer receiving CBDCA-based chemotherapy, but showed a prophylactic effect of nausea in female patients.
Asunto(s)
Protocolos de Quimioterapia Combinada Antineoplásica/efectos adversos , Medicamentos Herbarios Chinos/uso terapéutico , Náusea/prevención & control , Anciano , Anciano de 80 o más Años , Antieméticos/uso terapéutico , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapéutico , Carboplatino/administración & dosificación , Carboplatino/efectos adversos , Dexametasona/uso terapéutico , Femenino , Humanos , Masculino , Persona de Mediana Edad , Náusea/inducido químicamente , Náusea/tratamiento farmacológico , Estudios RetrospectivosRESUMEN
BACKGROUND: The outcome of anti-tuberculosis treatment varies according to patient factors. OBJECTIVE: To retrospectively identify risks related to the extension of time to negative sputum culture (Tn) and to determine their clinical significance. DESIGN: Patients with bacilli susceptible to isoniazid and rifampicin who received initial standard treatment without cessation were recruited into the study. A total of 630 consecutive in-patients were included in the risk development analysis (development cohort) and another 611 consecutive in-patients in the risk validation analysis (validation cohort). RESULTS: Univariate analysis showed that Tn was related to sex, body mass index (BMI), white blood cell count (WBC), serum albumin, fasting blood sugar, haemoglobin A1c, C-reactive protein and total cholesterol levels and sputum smear positivity (SSP). Multivariate analysis showed that BMI, WBC and SSP were significant risk factors related to extended Tn. Optimal cut-offs of BMI and WBC for predicting good (Tn < 46 days) and poor responders (Tn ⩾ 46 days) according to each risk were determined by receiver operating characteristics analysis. Risks were verified with the validation cohort. Tn increased according to the number of risks; the median Tn for patients with three risks was 21 days longer than that of patients with none. CONCLUSION: The nutritional state of a TB patient can be used to predict Tn.
Asunto(s)
Antituberculosos/uso terapéutico , Mycobacterium tuberculosis/aislamiento & purificación , Estado Nutricional , Esputo/microbiología , Tuberculosis/tratamiento farmacológico , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Biomarcadores/análisis , Índice de Masa Corporal , Proteína C-Reactiva/análisis , Femenino , Intolerancia a la Glucosa , Humanos , Isoniazida/uso terapéutico , Japón , Leucocitos , Masculino , Persona de Mediana Edad , Análisis Multivariante , Estudios Retrospectivos , Rifampin/uso terapéutico , Factores de Riesgo , Adulto JovenAsunto(s)
Antineoplásicos/efectos adversos , Carcinoma Hepatocelular/tratamiento farmacológico , Hemorragia Gastrointestinal/inducido químicamente , Neoplasias Hepáticas/tratamiento farmacológico , Niacinamida/análogos & derivados , Compuestos de Fenilurea/efectos adversos , Anciano de 80 o más Años , Endoscopía Capsular , Femenino , Hemorragia Gastrointestinal/diagnóstico , Humanos , Intestino Delgado , Niacinamida/efectos adversos , SorafenibRESUMEN
BACKGROUND AND OBJECTIVE: Antimicrobial photodynamic therapy (a-PDT) using a combination of red-colored laser/light-emitting diode (LED) and blue dye has been employed for periodontal therapy and the antimicrobial effect seems promising. Blue light, which has favorable wavelength properties, would be more effective as a light source for a-PDT because blue light itself possesses an antimicrobial effect. This study aimed to investigate the effect of a-PDT using a novel combination of high-power blue LED and red-dye agent on Porphyromonas gingivalis in vitro. MATERIAL AND METHODS: Porphyromonas gingivalis ATCC 33277 suspension was irradiated with blue LED (BL) (425-470 nm) or red LED (RL) (625-635 nm) at 30-90 J/cm(2) , or was mixed with erythrosine (ER), phloxine B (PB) or rose bengal (RB) with or without BL irradiation (30 J/cm(2) ). RL (30 J/cm(2) ) in combination with toluidine blue was employed as positive control. All the suspensions of P. gingivalis were serially diluted, plated and incubated anaerobically, and the numbers of colony-forming units (CFUs) were counted on day 7. RESULTS: BL irradiation at 60 and 90 J/cm(2) demonstrated a significant reduction in the numbers of CFUs. ER, PB and RB solutions at 160 µg/mL showed almost no or only a minimal reduction in the numbers of CFUs. BL at 30 J/cm(2) combined with ER, PB or RB at 160 µg/mL resulted in a log reduction of 0.9, 1.0 and 7.1, respectively, in the numbers of CFUs; 30 J/cm(2) BL with RB at 1.6, 16 and 160 µg/mL demonstrated a log reduction of 6.3, 8.0 and 5.5, respectively; and a log reduction of 5.2 was obtained after 30 J/cm(2) RL with 16 µg/mL TB. CONCLUSION: Within the limits of this study, BL was found to have an antimicrobial/growth-inhibiting effect on P. gingivalis, and a-PDT using a combination of BL and RB shows promise as a new technical modality for bacterial elimination in periodontal therapy.
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Antiinfecciosos/farmacología , Colorantes Fluorescentes/farmacología , Fotoquimioterapia/instrumentación , Fármacos Fotosensibilizantes/farmacología , Porphyromonas gingivalis/efectos de los fármacos , Carga Bacteriana/efectos de los fármacos , Técnicas Bacteriológicas , Colorantes/farmacología , Eritrosina/farmacología , Fluoresceínas/farmacología , Humanos , Fotoquimioterapia/métodos , Rosa Bengala/farmacología , Temperatura , Cloruro de Tolonio/farmacologíaRESUMEN
In conventional transmission electron microscopy, uranyl acetate staining is used to enhance the cellular components. However, uranyl acetate is considered a radioactive material that is very toxic if ingested or inhaled and subject to restrictions in many countries. In an attempt to introduce a substitute for uranyl acetate, we evaluated oolong tea extract (OTE) for staining of ultrathin sections. Tissue sections from normal rat liver representing an ideal model organ were processed according to a routine electron microscopic fixation and embedding procedure. Serial ultrathin sections were cut and processed with either routine double electron staining or 0.2% OTE staining for 30-40 min at room temperature followed by lead citrate staining (OTE staining method). Transmission electron microscopy observations revealed that all sub-cellular structures in hepatocytes were clearly visible with OTE staining and the quality of staining was highly compatible with those of routine double staining methods. It is suggested that OTE could be used as a non-radioactive and hazard-free substitute for uranyl acetate in transmission electron microscopy staining.
Asunto(s)
Hepatocitos/ultraestructura , Microscopía Electrónica de Transmisión/métodos , Extractos Vegetales/química , Coloración y Etiquetado/métodos , Té/química , Animales , Ácido Cítrico , Colorantes , Técnicas Histológicas , Plomo , RatasRESUMEN
OBJECTIVE: Gum arabic is a natural polysaccharide exudate from Acacia senegal and other related African species of Acacia. Gum arabic is considered to have an ability to enhance remineralization, because of its high concentration of Ca(2+). However, the caries preventive capacity of gum arabic has been scarcely investigated. We evaluated the cariostatic activities of gum arabic using histopathological methods to determine its effects on remineralization. DESIGN: Following incubation in demineralization solution, human third molars were exposed to 10 mg/ml of gum arabic, sodium fluoride at 1000 ppm (NaF), or double distilled water (DW, negative control), then subjected to demineralization-remineralization cycles. Before and after demineralization-remineralization cycles, contact microradiographs of each sample were taken and mineral distribution quantities were calculated. RESULTS: The remineralization ratio of the molars exposed to gum arabic was similar to that of those exposed to NaF, while the ratios of both were significantly greater than that of those exposed to DW. CONCLUSIONS: Gum arabic enhanced the remineralization of caries-like enamel lesions in vitro, suggesting its inhibitory effects towards dental caries.
Asunto(s)
Caries Dental/tratamiento farmacológico , Goma Arábiga/uso terapéutico , Incisivo/diagnóstico por imagen , Remineralización Dental , Humanos , Técnicas In Vitro , Incisivo/efectos de los fármacos , Microrradiografía , Fluoruro de Sodio/uso terapéutico , AguaRESUMEN
In vivo genotoxicity tests play a pivotal role in genotoxicity testing batteries. They are used both to determine if potential genotoxicity observed in vitro is realised in vivo and to detect any genotoxic carcinogens that are poorly detected in vitro. It is recognised that individual in vivo genotoxicity tests have limited sensitivity but good specificity. Thus, a positive result from the established in vivo assays is taken as strong evidence for genotoxic carcinogenicity of the compound tested. However, there is a growing body of evidence that compound-related disturbances in the physiology of the rodents used in these assays can result in increases in micronucleated cells in the bone marrow that are not related to the intrinsic genotoxicity of the compound under test. For rodent bone marrow or peripheral blood micronucleus tests, these disturbances include changes in core body temperature (hypothermia and hyperthermia) and increases in erythropoiesis following prior toxicity to erythroblasts or by direct stimulation of cell division in these cells. This paper reviews relevant data from the literature and also previously unpublished data obtained from a questionnaire devised by the IWGT working group. Regulatory implications of these findings are discussed and flow diagrams have been provided to aid in interpretation and decision-making when such changes in physiology are suspected.
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Células de la Médula Ósea/efectos de los fármacos , Pruebas de Mutagenicidad/métodos , Mutágenos/toxicidad , Compuestos de Anilina/toxicidad , Animales , Temperatura Corporal , Eritropoyetina/genética , Eritropoyetina/toxicidad , Guías como Asunto , Hipertermia Inducida , Pruebas de Micronúcleos , Naftoquinonas/toxicidad , Fenol/toxicidad , Fenilhidrazinas/toxicidad , Piridinas/toxicidad , Reserpina/toxicidad , Roedores , Sensibilidad y Especificidad , Triazoles/toxicidadRESUMEN
We report a 61-year-old man with aplastic anemia who underwent successful off-pump coronary artery bypass (OPCAB) after being admitted for angina pectoris. Coronary angiography showed severe stenosis of the left main coronary artery. Preoperative WBC was 2,200/microl, neutrophil 704/microl, Hb 8.1g/dl, and PLT 16,000/microl. We conducted OPCAB on double vessels using left internal thoracic and radial artery grafts. Thirty units of platelets were transfused intraoperatively with little perioperaive hemorrhage. Because of high grade fever, we injected 150 microg granulocyte colony-stimulating factor (G-CSF) every 3 days postoperatively to prevent major infection. The combination of appropriate perioperative management and OPCAB yielded an effective result for a patient with severe hematological disorders causing pancytopenia.
Asunto(s)
Anemia Aplásica/complicaciones , Angina de Pecho/cirugía , Puente de Arteria Coronaria Off-Pump , Angina de Pecho/diagnóstico por imagen , Transfusión de Sangre Autóloga , Angiografía Coronaria , Puente de Arteria Coronaria Off-Pump/métodos , Humanos , Masculino , Persona de Mediana Edad , Pancitopenia/etiología , Pancitopenia/terapia , Transfusión de PlaquetasRESUMEN
This study was designed to evaluate and characterize any subchronic toxicity of a new type of gum arabic (SUPER GUM [Acacia(sen)SUPER GUM]), a naturally processed polysaccharide exudate from gum acacia trees (Acacia senegal), when administered to both sexes of F344 rats at dietary levels of 0 (control), 1.25%, 2.5%, and 5.0% (10 rats/sex/group). During the study, the treatment had no effects on clinical signs, survival, body weights, and food and water consumption, or on findings of urinalysis, ophthalmology, hematology, or blood biochemistry. Gross pathology and histopathology exhibited no differences of toxicological significance between control and treated rats. Increased relative cecum (filled) weights, evident in both sexes of 5.0% group and females of 1.25% and 2.5% groups, were considered to be a physiological adaptation. Thus, the results indicated the toxic level of SUPER GUM to be more than 5.0%, and the no observed adverse effect level (NOAEL) was concluded to be 5.0% (3,117 mg/kg body weights/day for males, and 3,296 mg/kg body weights/day for males) from the present study.
Asunto(s)
Fabaceae , Aditivos Alimentarios/toxicidad , Goma Arábiga/toxicidad , Fitoterapia , Administración Oral , Animales , Femenino , Aditivos Alimentarios/administración & dosificación , Goma Arábiga/administración & dosificación , Masculino , Ratas , Ratas Endogámicas F344RESUMEN
PURPOSE: The treatment of rectal prolapse in children is controversial. We report the results of injection sclerotherapy in children using phenol in almond oil (PAO) and discuss the occurrence of complications after the injection of PAO. METHODS: Nine children with rectal prolapse, aged from 2 years and 6 months to 14 years, were treated by PAO injection sclerotherapy between 1993 and 2000. The outcome of PAO injection sclerotherapy and the presence of complications were investigated from the point of anorectal function using anorectal manometry. RESULTS: All of the nine patients were cured after one to three injections without any complications. The manometric study showed that normal anorectal reflex and other parameters of the anorectum were found after injection sclerotherapy. Two of the 4 who had complained of constipation no longer had constipation after the therapy. CONCLUSIONS: PAO injection sclerotherapy is simple and should be recommended as a first method of treatment for rectal prolapse in children. PAO as a sclerosing agent did not cause any complications.
Asunto(s)
Fenol/administración & dosificación , Prolapso Rectal/terapia , Soluciones Esclerosantes/administración & dosificación , Escleroterapia/métodos , Adolescente , Niño , Preescolar , Femenino , Humanos , Masculino , Manometría , Aceites de Plantas , Prolapso Rectal/fisiopatologíaRESUMEN
We investigated acute secondary degeneration in the thalamus following a cerebral infarct in 21 patients with an infarct in the territory of the middle cerebral artery, using serial MRI at various time after the stroke. Secondary degeneration in the ventral nuclei of the thalamus was seen as regions of slightly low signal on proton-density and/or T2-weighted images, mostly obtained a few weeks after the onset. An area of slightly high signal was observed in the dorsomedial nucleus of the thalamus on T2-weighted images about 6 weeks after the onset. Damage to the superior and anterior thalamic radiation caused degeneration in the ventral and dorsomedial nucleus, respectively. Thus, the time of detection and the abnormalities seen on MRI in secondary degeneration vary depending upon which area of the thalamus is involved. The mechanism underlying the degeneration is therefore also likely to differ in these areas.
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Infarto de la Arteria Cerebral Media/patología , Imagen por Resonancia Magnética , Tálamo/patología , Anciano , Anciano de 80 o más Años , Femenino , Humanos , Infarto de la Arteria Cerebral Media/fisiopatología , Masculino , Persona de Mediana Edad , Degeneración Nerviosa/patología , Estudios RetrospectivosRESUMEN
Pre-adipocyte factor-1 (pref-1), originally identified in mouse 3T3-L1 pre-adipocytes, is known to play a key role in inhibiting the adipose conversion. As a first step to study the involvement of the PREF1 gene in intramuscular adipose tissue development in cattle, which has an economic importance for beef cattle, we have isolated the bovine PREF1 cDNA and genomic clones and characterized expression in abdominal adipose and promoter region of the bovine PREF1 gene. We have detected two bovine PREF1 splice-isoforms, PREF1C2 and PREF1A, in abdominal fat tissue. The RT-PCR experiment revealed that the two isoforms are identified in neonatal, but no in adult abdominal fat tissue, suggesting age-dependent suppression of the bovine PREF1 gene expression in the form of PREF1C2 and PREF1A in abdominal fat tissue. By mapping the regulatory region of this gene, we have shown that at least two regions within 1121 bp upstream of putative transcription start site are sufficient to confer promoter activity, when accompanied by a short region including the transcription start site. The chromosomal location of the PREF1 gene was determined by fluorescence in situ hybridization (FISH). The PREF1 gene locates on bovine chromosome 21q24.
Asunto(s)
Proteínas de la Membrana/genética , Proteínas Represoras/genética , Células 3T3 , Animales , Proteínas de Unión al Calcio , Bovinos , Mapeo Cromosómico , Clonación Molecular , ADN Complementario/aislamiento & purificación , Expresión Génica , Péptidos y Proteínas de Señalización Intercelular , Ratones , Regiones Promotoras Genéticas , Análisis de Secuencia de ADNRESUMEN
Complementary DNA encoding a DNA-binding protein, designated PLATZ1 (plant AT-rich sequence- and zinc-binding protein 1), was isolated from peas. The amino acid sequence of the protein is similar to those of other uncharacterized proteins predicted from the genome sequences of higher plants. However, no paralogous sequences have been found outside the plant kingdom. Multiple alignments among these paralogous proteins show that several cysteine and histidine residues are invariant, suggesting that these proteins are a novel class of zinc-dependent DNA-binding proteins with two distantly located regions, C-x(2)-H-x(11)-C-x(2)-C-x((4-5))-C-x(2)-C-x((3-7))-H-x(2)-H and C-x(2)-C-x((10-11))-C-x(3)-C. In an electrophoretic mobility shift assay, the zinc chelator 1,10-o-phenanthroline inhibited DNA binding, and two distant zinc-binding regions were required for DNA binding. A protein blot with (65)ZnCl(2) showed that both regions are required for zinc-binding activity. The PLATZ1 protein non-specifically binds to A/T-rich sequences, including the upstream region of the pea GTPase pra2 and plastocyanin petE genes. Expression of the PLATZ1 repressed those of the reporter constructs containing the coding sequence of luciferase gene driven by the cauliflower mosaic virus (CaMV) 35S90 promoter fused to the tandem repeat of the A/T-rich sequences. These results indicate that PLATZ1 is a novel class of plant-specific zinc-dependent DNA-binding protein responsible for A/T-rich sequence-mediated transcriptional repression.
Asunto(s)
Secuencia Rica en At , Proteínas de Unión al ADN/genética , Proteínas de Unión al ADN/metabolismo , Regulación de la Expresión Génica de las Plantas , Proteínas de Plantas/metabolismo , Zinc/farmacología , Secuencia de Aminoácidos , Secuencia de Bases , Sitios de Unión , Clonación Molecular , ADN de Plantas/metabolismo , Proteínas de Unión al ADN/fisiología , Datos de Secuencia Molecular , Pisum sativum/genética , Pisum sativum/metabolismo , Filogenia , Proteínas de Plantas/genética , Proteínas de Plantas/fisiología , ARN de Planta/biosíntesis , Proteínas Represoras/genética , Proteínas Represoras/fisiología , Homología de Secuencia de Aminoácido , Transcripción GenéticaRESUMEN
A new hereditary disease characterised by renal failure, poor growth and long hooves in Japanese Black cattle (wagyu) has been recognised in a region of central Japan since 1990. The number of calves affected has increased gradually, with the incidence reaching 17 of 485 (3.51 per cent) in 1995. Almost all the calves were slightly undersized at birth, and repeatedly had diarrhoea during the neonatal period. They began to show signs of growth retardation with proportional body and elongation of the hooves from about two to five months of age, but they had an almost normal or only slightly decreased appetite. The concentrations of urea nitrogen, creatinine and inorganic phosphorus in serum were high, and the affected calves excreted diluted urine frequently. Among 25 cases, the urine of 21 contained occult blood, 24 contained protein and two contained glucose. In 29 calves observed for 30 to 130 days, the course of the disease varied; in 21 of them it remained unchanged, six became gradually worse and two became severely debilitated and died. The disease was diagnosed as renal tubular dysplasia by histopathological examination.
Asunto(s)
Enfermedades de los Bovinos/fisiopatología , Enfermedades Renales/veterinaria , Túbulos Renales/fisiopatología , Animales , Animales Recién Nacidos , Apetito , Peso al Nacer , Nitrógeno de la Urea Sanguínea , Bovinos , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/genética , Creatinina/sangre , Diarrea , Brotes de Enfermedades/estadística & datos numéricos , Brotes de Enfermedades/veterinaria , Resultado Fatal , Femenino , Enfermedades del Pie/fisiopatología , Enfermedades del Pie/veterinaria , Pezuñas y Garras/crecimiento & desarrollo , Pezuñas y Garras/patología , Incidencia , Japón/epidemiología , Enfermedades Renales/epidemiología , Enfermedades Renales/genética , Enfermedades Renales/fisiopatología , Túbulos Renales/patología , Masculino , Fósforo/sangre , UrinálisisRESUMEN
Curcuma drugs have been used discriminatingly for invigorating blood circulation, promoting digestion, and as a cholagogic in China. However, there is confusion about the drug's botanical origins and clinical uses because of morphological similarity of Curcuma plants and drugs. In order to develop an ultimate identification, molecular analysis based on 18S rRNA gene and trnK gene sequences were performed on 6 Curcuma species used medicinally in China and Japan. The 18S rRNA gene sequences were found to be of 1810 bps in length. In comparison with the common sequence of C. longa, C. phaeocaulis, C. wenyujin and C. aromatica, that of C. kwangsiensis had one base substitution, and the same base difference was observed between the Chinese and the Japanese populations of C. zedoaria. The trnK gene sequences were found to span 2698-2705 bps. There were base substitutions, small deletions or insertions at some sites between the trnK coding region and matK region among each species. Based on the base substitutions, C. zedoaria and C. kwangsiensis specimens were divided into two groups, respectively. An identical sequence was detected in C. phaeocaulis and in the Chinese population of C. zedoaria, as well as in the Japanese population of C. zedoaria and in one group of C. kwangsiensis with a purple-colored band in leaves. New taxonomic information to be used for authenticating Curcuma drugs was obtained.
Asunto(s)
Curcuma/genética , Medicamentos Herbarios Chinos/análisis , ARN Ribosómico 18S/análisis , ARN Ribosómico 18S/genética , Análisis de Secuencia de ADN/métodos , Medicamentos Herbarios Chinos/aislamiento & purificación , Japón , Filogenia , Estructuras de las Plantas/química , Estructuras de las Plantas/genética , ARN Ribosómico 18S/aislamiento & purificación , Análisis de Secuencia de ADN/estadística & datos numéricos , Zingiberaceae/genéticaRESUMEN
For direct observation of the surface structures of soft-tissue specimens, we examined rat tracheal tissue in a low-vacuum scanning electron microscope (SEM) equipped with a cooling stage. In specimens fixed with glutaraldehyde and osmium tetroxide, back-scattered electron images of the surface structure could not be clearly observed in the low-vacuum SEM because of the disruption of fine structures and a low signal-to-noise (S/N) ratio. Processing of the specimens in 70% ethanol resulted in marked shrinkage, in contrast to results when processing in 30% ethanol. To overcome these problems, the trachea was initially fixed in 2.5% glutaraldehyde (0.1 M phosphate buffer pH 7.4), treated with a mixture of 0.2% oolong tea extract (OTE) and 2.5% glutaraldehyde, and postfixed in 1% osmium tetroxide. The sample was immersed in 30% ethanol and examined in a chilled SEM at -10 degrees C. The luminal coutour of the tracheal epithelial cells was clearly observed because of the decrease in shrinkage. Cilia of ciliated cells and microvilli of nonciliated cells were also clearly observed. These specimens also showed a high S/N ratio, thus allowing the observation of samples without the need for complete dehydration, critical-point drying, or metal coating. This OTE-incorporated conductive staining method is simple and rapid, and should prove to be highly useful for rapid SEM analyses of biological specimens.
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Microscopía Electrónica de Rastreo/métodos , Extractos Vegetales/farmacología , Té , Tráquea/ultraestructura , Animales , Frío , Masculino , Ratas , Ratas Wistar , Coloración y EtiquetadoRESUMEN
Bra r 1 encodes a Ca2+-binding protein specifically expressed in anthers of Brassica rapa. In this study, we isolated a genomic clone of Bra r 1 and found sequences similar to Pollen Box core motifs and LAT56/59 box, pollen-specific cis-acting element, in the 5' upstream region of Bra r 1. Reporter gene fusion revealed that the Bra r 1 promoter directs male gametophytic expression in Nicotiana tabacum, Arabidopsis thaliana and B. napus, showing strong expression in mature pollen grains similar to that of endogenous Bra r 1. Genomic DNA of Bra r 1 was introduced into tobacco plants and the highest accumulation of Bra r 1 protein was observed in mature pollen in the same manner as reporter gene expression. Using in vitro-germinated pollen tubes of transgenic tobacco, we firstly demonstrated the subcellular localization of Bra r 1 in pollen tubes. Bra r 1 protein was distributed throughout the pollen tube of transgenic tobacco and slightly intense signals of Bra r 1 were observed in the tip region. In long-germinated pollen tubes, Bra r 1 was detected only in the cytoplasmic compartments while no signals were observed in the empty part of the pollen tube, indicating that cytoplasmic movement toward the tube tip is accompanied by Bra r 1. Hence, we suggest that Bra r 1 is involved in pollen germination and pollen tube growth.
Asunto(s)
Arabidopsis/genética , Brassica/genética , Proteínas de Unión al Calcio/genética , Nicotiana/fisiología , Proteínas de Plantas , Plantas Tóxicas , Polen/fisiología , Regiones Promotoras Genéticas , Secuencia de Aminoácidos , Secuencia de Bases , Proteínas de Unión al Calcio/análisis , Proteínas de Unión al Calcio/química , Datos de Secuencia Molecular , Plantas Modificadas Genéticamente , Especificidad de la Especie , TATA Box , Nicotiana/genéticaRESUMEN
Inhibitors of angiotensin converting enzyme (ACE) have been developed recently for therapeutic purposes in hypertension and ischemic cardiovascular diseases. Ogiku et al. reported that one such inhibitor, imidapril, significantly prolonged survival in stroke-prone spontaneously hypertensive rats (SHRSP). The present study was designed to investigate the effect of imidapril on cerebral blood vessels in SHRSP to clarify role of the ACE inhibitor in mechanisms of cerebral thrombosis and stroke. Imidapril was administered orally at 1.0 and 5.0 mg/kg/day for 3 weeks from the age of 7 weeks, and was shown to prevent the usual increase in blood pressure seen in these animals. It also delayed He-Ne laser-induced cerebral thrombosis and increased significantly the plasma concentration of nitric oxide metabolites (NO2/NO3). To confirm the association between nitric oxide (NO) and these effects of imidapril, an inhibitor of nitric oxide synthase, N(G)-nitro-L-arginine methyl ester hydrochloride (L-NAME) was dissolved in drinking water and administered to the animals for 3 weeks. Four of six rats died from stroke when L-NAME was given alone. When imidapril (5.0 mg/kg/day) was administered with L-NAME, however, the animals showed no signs or symptoms of stroke. In these instances, therefore, the concurrent administration of L-NAME with imidapril reversed significantly the effects of imidapril. Intravenous injection of imidaprilat (100 microg/kg), an active metabolite of imidapril, also decreased blood pressure significantly and increased the plasma levels of NO2/NO3 after 5 min. Moreover, imidaprilat enlarged arteriolar diameters and caused an increase in red cell velocity and mean blood flow in pial arterioles after 15 min. The results strongly suggested that imidapril protects cerebral vessels in SHRSP by elevating the release of NO, thereby improving the cerebral circulation and reducing the tendency to thrombosis and stroke.
Asunto(s)
Inhibidores de la Enzima Convertidora de Angiotensina/uso terapéutico , Antihipertensivos/uso terapéutico , Imidazoles/uso terapéutico , Imidazolidinas , Trombosis Intracraneal/prevención & control , Inhibidores de la Enzima Convertidora de Angiotensina/farmacología , Animales , Antihipertensivos/farmacología , Arteriolas/ultraestructura , Arterias Cerebrales/ultraestructura , Evaluación Preclínica de Medicamentos , Femenino , Predisposición Genética a la Enfermedad , Hemorreología/efectos de los fármacos , Hipertensión/genética , Hipertensión/prevención & control , Imidazoles/antagonistas & inhibidores , Imidazoles/farmacología , Trombosis Intracraneal/tratamiento farmacológico , Trombosis Intracraneal/genética , Rayos Láser/efectos adversos , Masculino , Microcirculación/efectos de los fármacos , NG-Nitroarginina Metil Éster/toxicidad , Óxido Nítrico/fisiología , Óxido Nítrico Sintasa/antagonistas & inhibidores , Ratas , Ratas Endogámicas SHRRESUMEN
Although L-3,4-dihydroxyphenylalanine (L-DOPA) is claimed to be a neurotransmitter in the central nervous system (CNS), receptor or transporter molecules for L-DOPA have not been determined. In an attempt to identify a transporter for L-DOPA, we examined whether or not an active and high affinity L-DOPA transport system is expressed in Xenopus laevis oocytes injected with poly A(+) RNA prepared from several tissues. Among the poly A(+) RNAs tested, rabbit intestinal epithelium poly A(+) RNA gave the highest transport activity for L-[(14)C]DOPA in the oocytes. The uptake was approximately five times higher than that of water-injected oocytes, and was partially Na(+)-dependent. L-Tyrosine, L-phenylalanine, L-leucine and L-lysine inhibited this transport activity, whereas D-DOPA, dopamine, glutamate and L-DOPA cyclohexylester, an L-DOPA antagonist did not affect this transport. Coinjection of an antisense cRNA, as well as oligonucleotide complementary to rabbit rBAT (NBAT) cDNA almost completely inhibited the uptake of L-[(14)C]DOPA in the oocytes. On the other hand, an antisense cRNA of rabbit 4F2hc barely affected this L-[(14)C]DOPA uptake activity. rBAT was thus responsible for the L-[(14)C]DOPA uptake activity expressed in X. laevis oocytes injected with poly A(+) RNA from rabbit intestinal epithelium. As rBAT is localized at the target regions of L-DOPA in the CNS, rBAT might be one of the components involved in L-DOPAergic neurotransmission.