RESUMEN

OBJECTIVES: To investigate the allergen-induced messenger RNA (mRNA) expression of interleukin (IL) 4, IL-5 and interferon gamma (IFN-gamma) in peripheral blood mononuclear cells from individuals sensitized by Japanese cedar (Cryptomeria japonica) pollens, and to elucidate the clinical role of IL-4, IL-5, and IFN-gamma in the allergen sensitization and clinical manifestation of allergic disorders. DESIGN: This study included 30 patients sensitized to the pollen and 14 nonatopic healthy volunteers. Peripheral blood mononuclear cells (1.0 x 10(6) cells/mL) of each individual were cultured at 37 degrees C for 24 hours in the presence of 10 microg/mL of Cry j 1, a major allergen of the pollens. Total cellular RNA was extracted from the peripheral blood mononuclear cells, and IL-4, IL-5, and IFN-gamma mRNA expression was determined with a reverse transcriptase polymerase chain reaction. RESULTS: From the results of a survey of symptom diary cards and interviews regarding nasal symptoms during the pollen season in 1998, we found that 20 patients (symptomatic group), but not 10 patients (asymptomatic group), had typical symptoms of seasonal allergic rhinitis. Interleukin 4 mRNA was not expressed in the nonatopic subjects but was expressed in 9 asymptomatic patients and in 17 symptomatic patients. Interleukin 5 mRNA was exclusively expressed in the symptomatic patients. Interferon gamma mRNA expression did not differ significantly among the nonatopic subjects, asymptomatic patients, and symptomatic patients. CONCLUSIONS: This study has clearly highlighted an interesting and new concept that IL-4 is implicated in allergen sensitization but not in clinical manifestation, and that IL-5 may not be a feature of atopy in itself but seems to be a hallmark of clinical manifestation of ongoing atopic diseases.

Asunto(s)

Alérgenos/efectos adversos , Expresión Génica/genética , Interferón gamma/genética , Interleucina-4/genética , Interleucina-5/genética , Rinitis Alérgica Estacional/inmunología , Adolescente , Adulto , Cromatografía de Afinidad/métodos , Femenino , Humanos , Leucocitos Mononucleares/metabolismo , Masculino , Persona de Mediana Edad , Polen/genética , ARN Mensajero/genética , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa/métodos , Rinitis Alérgica Estacional/genética

RESUMEN

The primary aim of this study was to investigate whether the allergen-induced synthesis of cytokines by peripheral blood mononuclear cells (PBMCs) obtained immediately before the pollen season could predict the clinical efficacy of immunotherapy during the following pollen season. PBMCs (1.0 x 106 cells/ml) were obtained from 17 nonatopic subjects and from 60 patients receiving immunotherapy for seasonal allergic rhinitis (caused by Japanese cedar pollens) immediately before the pollen season of 1998, and were cultured for 24 h in the presence of 10 mg/ml of Cry j 1, a major allergen of Japanese cedar pollens, at 37 degrees C in a fully humidified 5% CO2 atmosphere. Total cellular RNA was extracted from the PBMCs, and the allergen-induced interleukin (IL)-4, IL-5 and interferon-gamma (IFN-gamma) mRNA expression was determined using a reverse transcription-polymerase chain reaction. According to the nasal symptoms during the pollen season of 1998, the 60 patients on immunotherapy were divided into 36 good responders (who had no nasal symptoms and no requirement for rescue medications) and 24 poor responders who needed rescue medications to control nasal symptoms. Neither IL-4 mRNA nor IL-5 mRNA was expressed in any of the 17 nonatopic individuals. By contrast, IL-4 mRNA was expressed in 26 good responders and in 22 poor responders, and IL-5 mRNA was expressed in eight good responders (22.2%) and in 23 poor responders (95.8%). IFN-gamma mRNA was expressed in four nonatopic subjects, in nine good responders and in seven poor responders. The expression of IFN-gamma did not differ significantly among the nonatopic subjects, the good responders and the poor responders. The mRNA expression of IL-5 (P < 0.0001), but not of IL-4 (P = 0.0999) and IFN-gamma (P = 0. 7713), differed significantly between the good and poor responders. Therefore, our study has highlighted that positive expression of IL-5 mRNA in PBMCs sampled immediately before the pollen season could be predictive of a poor clinical outcome of immunotherapy during the following pollen season and that the down-regulation of IL-5 mRNA expression in PBMCs could be an important mechanism of pollen immunotherapy related to the clinical efficacy.

Asunto(s)

Alérgenos/inmunología , Interferón gamma/biosíntesis , Interleucina-4/biosíntesis , Interleucina-5/biosíntesis , Leucocitos Mononucleares/inmunología , Polen/inmunología , Rinitis Alérgica Estacional/inmunología , Actinas/genética , Adolescente , Adulto , Antígenos de Plantas , Femenino , Expresión Génica , Humanos , Inmunoglobulina E/sangre , Inmunoglobulina E/inmunología , Interferón gamma/genética , Interleucina-4/genética , Interleucina-5/genética , Masculino , Persona de Mediana Edad , Proteínas de Plantas/inmunología , Valor Predictivo de las Pruebas , ARN Mensajero , Rinitis Alérgica Estacional/genética , Rinitis Alérgica Estacional/terapia , Estaciones del Año , Resultado del Tratamiento

RESUMEN

Urinary excretion of uric acid was found to be extremely low in a 58-year-old female patient with alcaptonuria. This was due to interference with the uricase-peroxidase method used, because analysis using high-performance liquid chromatography (HPLC) showed a normal urinary concentration of uric acid. In vitro experiments demonstrated that a high concentration of homogentisic acid in the patient's urine inhibited the peroxidase reaction, possibly due to inhibition of the colour development of 3-methyl-N-ethyl-N-(beta-hydroxyethyl)aniline (MEHA) and 4-aminoantipyrine, via the peroxidase reaction. A homogentisic acid concentration equivalent to that in plasma did not affect the uricase-peroxidase reaction. This result suggests that any assay based on a peroxidase method is affected by a high urinary concentration of homogentisic acid in patients with alcaptonuria.

Asunto(s)

Alcaptonuria/orina , Ácido Homogentísico/orina , Ácido Úrico/orina , Alcaptonuria/complicaciones , Femenino , Humanos , Persona de Mediana Edad , Peroxidasa/metabolismo , Urato Oxidasa/metabolismo

Asunto(s)

Arteriosclerosis/epidemiología , Gota/sangre , Gota/complicaciones , Factores de Edad , Consumo de Bebidas Alcohólicas , Apolipoproteínas/sangre , Arteriosclerosis/complicaciones , Presión Sanguínea , Colesterol/sangre , Gota/fisiopatología , Humanos , Japón , Lipoproteínas/sangre , Masculino , Persona de Mediana Edad , Factores de Riesgo , Triglicéridos/sangre

RESUMEN

This study was designed to compare the clinical outcome of prolonged immunotherapy for perennial allergic rhinitis with that of pharmacological treatment. Patients with perennial allergic rhinitis due to Dermatophagoides farinae (D. farinae) were divided into two groups; a pharmacotherapy group and an immunotherapy group. The pharmacotherapy group was treated with conventional pharmacological treatment using antihistamine tablets and topical steroid sprays and the immunotherapy group was treated with D. farinae extracts for 5 successive years. None of symptom scores at enrollment differed significantly between the groups. At 6 months and 1 year after the start of treatment the rate of decrease in each score was significantly greater in the pharmacotherapy group than in the immunotherapy group. The rate of decrease in sneezing scores, but not in the other scores, at 2 years after the start of treatment was also greater in the pharmacotherapy group than in the immunotherapy group. However, at 3 years the rate of decrease in any of the scores did not differ significantly between groups. The differences between the groups became clear-cut again after 5 years of treatment, when the rate of decrease in all of the scores was significantly greater in the immunotherapy group than in the pharmacotherapy group. Therefore, short-term treatment with pharmacological agents is probably superior to immunotherapy but, in the long-term, immunotherapy is apparently superior to pharmacological treatment with respect to clinical efficacy. In addition, prolonged immunotherapy provided long-term clinical efficacy and might provide a long-standing cure even after discontinuation of the therapy. In a questionnaire interview, approximately half of patients were very satisfied with prolonged immunotherapy and three-quarters were fairly satisfied or more. Additionally, the magnitude of improvement in nasal stuffiness contributed significantly and exclusively to the patient evaluation of immunotherapy. We propose that prolonged immunotherapy is never inferior to anti-allergenic pharmacological treatment and that it is possible to achieve long-term clinical efficacy or long-standing cure even after the discontinuation of immunotherapy, and that patients with perennial allergic rhinitis will be very satisfied with this prolonged therapeutic technique if nasal stuffiness is considerably alleviated.

Asunto(s)

Desensibilización Inmunológica , Rinitis Alérgica Perenne/terapia , Adolescente , Corticoesteroides/uso terapéutico , Adulto , Animales , Antígenos Dermatofagoides , Femenino , Estudios de Seguimiento , Glicoproteínas/inmunología , Antagonistas de los Receptores Histamínicos H1/uso terapéutico , Humanos , Inmunoglobulina E/sangre , Masculino , Persona de Mediana Edad , Ácaros/inmunología , Rinitis Alérgica Perenne/diagnóstico , Rinitis Alérgica Perenne/tratamiento farmacológico , Resultado del Tratamiento

RESUMEN

The most serious problem in practical immunotherapy is the risk of occasional, potentially life-threatening adverse systemic reactions. Elucidation of the incidence of, and possible risk factors for, systemic reactions would have a profound effect on the decision about how to manage allergic rhinitis. The aim of this retrospective study was to document the incidence and risk factors of adverse systemic reactions during immunotherapy using standardized Dermatophagoides farinae (D. farinae) extracts for perennial allergic rhinitis. This study included 386 patients (22,722 injections) with perennial allergic rhinitis who had received immunotherapy with standardized D. farinae extracts in our clinics for the past 5 years. The incidence of systemic reactions was 6.22% per patient and 0.12% per injection. The time of onset of systemic reactions ranged from 3 to 30 min (mean 11.3 min) after injections. Our study has demonstrated that asthma, atopic dermatitis and a high level of IgE in serum, but not a high level of specific IgE in serum, are important high risk factors that may induce severe adverse systemic reactions in patients who receive immunotherapy for perennial allergic rhinitis. The incidence of systemic reactions in those who had a high level of IgE (higher than 1000 U/ml) and asthma and/or atopic dermatitis was 66.67% (12/18) per patient. Conversely, the incidence of systemic reactions in those who had none of the risk factors was 1.64% per patient. Thus, the rate of systemic reactions is thought to be reduced by 75% if patients with high risk factors are strictly excluded from immunotherapy for allergic rhinitis.

Asunto(s)

Alérgenos/efectos adversos , Anafilaxia/etiología , Desensibilización Inmunológica/efectos adversos , Glicoproteínas/efectos adversos , Rinitis Alérgica Perenne/terapia , Alérgenos/uso terapéutico , Animales , Antígenos Dermatofagoides , Glicoproteínas/uso terapéutico , Humanos , Ácaros/inmunología , Estudios Retrospectivos , Factores de Riesgo

RESUMEN

The aim of this study was to determine whether there is a difference in the specific IgE response to pollen exposure between asymptomatic and symptomatic subjects who were already sensitized to Japanese cedar pollen. Sixty-four subjects with detectable serum levels of cedar pollen-specific IgE, which were apparent even before the pollen season in 1997, were enrolled in the study. Thirty-five had typical symptoms of seasonal allergic rhinitis during the pollen season in 1997 (symptomatic group) and the remainder had no seasonal aggravation during the pollen season in 1997 (asymptomatic group). Serum samples were collected twice from each subject, before and during the pollen season in 1997, to determine specific IgE by the lumiward immunoassay system. In both groups, the serum levels of specific IgE during the pollen season were significantly higher than those before the pollen season, and the rates of seasonal increase in specific IgE did not differ significantly between the groups. In conclusion, the specific IgE response during the pollen season is not a hallmark of clinical allergy and does not discriminate between symptomatic and asymptomatic individuals sensitive to the pollen.

Asunto(s)

Alérgenos/inmunología , Inmunoglobulina E/sangre , Polen/inmunología , Rinitis Alérgica Estacional/inmunología , Estaciones del Año , Especificidad de Anticuerpos , Humanos , Japón , Árboles

RESUMEN

We examined the usefulness of positron emission tomography (PET) using fluorine-18 deoxyglucose (FDG) in determining the therapeutic effects of irradiation and chemotherapy on head and neck malignant tumors. Twenty-two patients with head and neck lesions who underwent histological examinations were studied. Squamous cell carcinoma was histologically diagnosed in all cases. Sixteen of them underwent radiotherapy with approximately 40 Gy in combination with carboplatin therapy. The remaining 6 patients underwent radiotherapy alone. After these treatments, 11 underwent surgery. For PET study, each patient was injected with intravenous FDG 185-370 MBq. We evaluated the degree of FDG accumulation using scanned images taken 40-55 min after the injection. We measured the standardized uptake value (SUV), a semiquantative evaluation, ROI activity divided by the dosage per weight of each patient. FDG-PET, CT and MRI were performed twice for each patient, before and after treatment. FDG uptake, but not the tumor size in CT or MRI, was significantly reduced in each patient after the treatment. Therefore, our findings have clearly demonstrated that FDG-PET provides for more valuable therapeutic outcomes than conventional imaging such as CT and MRI. FDG-PET should thus provide a new dimension in the management of head and neck malignant tumors.

Asunto(s)

Carcinoma de Células Escamosas/diagnóstico por imagen , Carcinoma de Células Escamosas/radioterapia , Fluorodesoxiglucosa F18 , Neoplasias de Cabeza y Cuello/diagnóstico por imagen , Neoplasias de Cabeza y Cuello/radioterapia , Radiofármacos , Tomografía Computarizada de Emisión , Humanos

RESUMEN

An assay for human plasma xanthine oxidase activity was developed with pterin as the substrate and the separation of product (isoxanthopterin) by high-performance liquid chromatography with a fluorescence detector. The reaction mixture consists of 60 microliters of plasma and 240 microliters of 0.2 M Tris-HCl buffer (pH 9.0) containing 113 microM pterin. With this assay, the activity of plasma xanthine oxidase could be easily determined despite its low activity. As a result, it could be demonstrated that the intravenous administration of heparin or the oral administration of ethanol did not increase plasma xanthine oxidase activity in normal subjects, and also that plasma xanthine oxidase activity was higher in patients with hepatitis C virus infection than in healthy subjects or patients with gout. In addition, a single patient with von Gierke's disease showed a marked increase in the plasma activity of this enzyme, relative to that apparent in normal subjects.

Asunto(s)

Cromatografía Líquida de Alta Presión/métodos , Xantina Oxidasa/sangre , Etanol/farmacología , Enfermedad del Almacenamiento de Glucógeno Tipo I/enzimología , Gota/enzimología , Heparina/farmacología , Hepatitis C/enzimología , Humanos , Concentración de Iones de Hidrógeno , Hipoxantina/farmacología , Pterinas/metabolismo , Espectrometría de Fluorescencia , Ácido Úrico/farmacología , Xantina , Xantinas/farmacología , Xantopterina/análisis , Xantopterina/metabolismo

Asunto(s)

Benzbromarona , Síndrome de Fanconi/fisiopatología , Riñón/fisiopatología , Pirazinamida , Ácido Úrico/metabolismo , Uricosúricos , Adolescente , Benzbromarona/farmacocinética , Humanos , Masculino , Pirazinamida/farmacocinética , Factores de Tiempo , Uricosúricos/farmacocinética

RESUMEN

A high-performance liquid chromatographic method was developed for the determination of plasma purine nucleoside phosphorylase activity. In this method, the reaction mixture consisted of 15 microliters of plasma and 285 microliters of 50 mM phosphate buffer (pH 7.4) containing 3.8 mM inosine and 0.15 mM 2-(3-cyano-4-isobutoxyphenyl)-4-methyl-5-thiazolecarboxylic acid (strong xanthine oxidase inhibitor). After the reaction, the hypoxanthine produced was monitored to express plasma purine nucleoside phosphorylase activity. By this method, the activity of purine nucleoside phosphorylase was easily determined even with a small-volume plasma sample and despite its low activity in plasma. In addition, plasma purine nucleoside phosphorylase activity can be accurately determined even if the plasma is turbid. As a result, we were able to measure plasma purine nucleoside phosphorylase activity in patients with gout or asthma and healthy subjects, whereby it was demonstrated that plasma purine nucleoside phosphorylase activity was higher in patients with asthma than in either healthy subjects or patients with gout.

Asunto(s)

Cromatografía Líquida de Alta Presión/métodos , Hipoxantinas/análisis , Inosina/metabolismo , Purina-Nucleósido Fosforilasa/sangre , Adulto , Anciano , Asma/enzimología , Calibración , Febuxostat , Femenino , Gota/enzimología , Supresores de la Gota/metabolismo , Humanos , Hipoxantina , Hipoxantinas/metabolismo , Masculino , Persona de Mediana Edad , Purina-Nucleósido Fosforilasa/metabolismo , Tiazoles/metabolismo , Ácido Úrico/análisis , Ácido Úrico/metabolismo , Xantina Oxidasa/antagonistas & inhibidores , Xantina Oxidasa/metabolismo

Asunto(s)

Alopurinol/farmacología , Alopurinol/farmacocinética , Glucosa/farmacología , Riñón/fisiología , Oxipurinol/metabolismo , Ácido Úrico/metabolismo , Adulto , Glucosa/administración & dosificación , Humanos , Infusiones Intravenosas , Riñón/efectos de los fármacos , Masculino , Manitol/administración & dosificación , Manitol/farmacología , Valores de Referencia , Albúmina Sérica/metabolismo , Xantina , Xantinas/metabolismo

Asunto(s)

Aldehído Oxidorreductasas/metabolismo , Alopurinol/metabolismo , Hígado/enzimología , Pirazinamida/sangre , Aldehído Oxidasa , Animales , Biotransformación , Masculino , Oxidación-Reducción , Oxipurinol/sangre , Pirazinamida/análogos & derivados , Ratas , Ratas Wistar , Triazinas/farmacología , Xantina Deshidrogenasa/antagonistas & inhibidores , Xantina Oxidasa/antagonistas & inhibidores

Asunto(s)

Hipoxantinas/metabolismo , Xantina Oxidasa/deficiencia , Xantinas/metabolismo , 5'-Nucleotidasa/metabolismo , Adenina Fosforribosiltransferasa/metabolismo , Adenosina Desaminasa/metabolismo , Carcinoma Hepatocelular , Línea Celular , Medios de Cultivo , Citosol/enzimología , Guanina Desaminasa/metabolismo , Humanos , Hipoxantina , Hipoxantina Fosforribosiltransferasa/metabolismo , Hígado/enzimología , Neoplasias Hepáticas , Purina-Nucleósido Fosforilasa/metabolismo , Células Tumorales Cultivadas , Xantina , Xantina Oxidasa/metabolismo

Asunto(s)

Gota/enzimología , Lipasa/sangre , Lipoproteína Lipasa/sangre , Apolipoproteínas B/sangre , Índice de Masa Corporal , Colesterol/sangre , HDL-Colesterol/sangre , Estudios de Cohortes , Gota/sangre , Humanos , Hígado/enzimología , Masculino , Valores de Referencia , Triglicéridos/sangre , Ácido Úrico/sangre

RESUMEN

Allopurinol or pyrazinamide was administered to rats treated with BOF-4272 (a potent xanthine oxidase inhibitor) to investigate to what degree xanthine dehydrogenase participates in the oxidation of these agents. BOF-4272 markedly decreased the plasma concentration and the urinary excretion of both oxypurinol and 5-hydroxypyrazinamide. It also decreased the sum of the urinary excretion of allopurinol and oxypurinol and that of pyrazinamide and its metabolites, although it did not affect the sum of the plasma concentrations of allopurinol and oxypurinol at 105 min after administration of allopurinol or the plasma concentration of pyrazinamide during the period after the administration of pyrazinamide. These results suggested that BOF-4272 almost completely inhibited the oxidation of allopurinol and pyrazinamide and had some effect on the excretion and/or the tissue incorporation of these two compounds. Since the in vitro study demonstrated that BOF-4272 did not inhibit the activity of aldehyde oxidase, which oxidized both allopurinol to oxypurinol and pyrazinamide to 5-hydroxypyrazinamide, the results suggested that xanthine dehydrogenase was the more important enzyme in converting allopurinol to oxypurinol and pyrazinamide to 5-hydroxypyrazinamide.

Asunto(s)

Aldehído Oxidorreductasas/metabolismo , Alopurinol/metabolismo , Pirazinamida/metabolismo , Triazinas/farmacología , Xantina Deshidrogenasa/metabolismo , Aldehído Oxidasa , Alopurinol/orina , Animales , Hipoxantina Fosforribosiltransferasa/metabolismo , Masculino , Oxidación-Reducción , Oxipurinol/sangre , Oxipurinol/orina , Pirazinamida/sangre , Pirazinamida/orina , Ratas , Ratas Wistar , Triazinas/sangre , Xantina Deshidrogenasa/antagonistas & inhibidores

RESUMEN

To investigate whether or not DL-sodium lactate inhibits the renal excretion of purine bases and oxypurinol, we administered physiological saline containing 0.2 mol DL-sodium lactate to 7 normal subjects intravenously. DL-sodium lactate infusion decreased the urinary excretion and the fractional clearance of uric acid, xanthine and oxypurinol, but the fractional clearance of hypoxanthine was not affected. These results suggested that the implications of DL-sodium lactate-induced hyperuricemia must be considered in patients with gout on its long term and high dose administration, and that the implications of DL-sodium lactate-induced prolongation of half-life of oxypurinol must be considered in hyperuricemic patients treated with allopurinol. However, since the high dose and long term administration of DL-sodium lactate is clinically rare, the effect of DL-sodium lactate infusion on the urinary excretion of uric acid, xanthine and oxypurinol may not be clinically important.

Asunto(s)

Lactatos/farmacología , Oxipurinol/orina , Purinas/orina , Adulto , Alopurinol/farmacología , Humanos , Hipoxantina , Hipoxantinas/orina , Lactatos/sangre , Ácido Láctico , Masculino , Albúmina Sérica/análisis , Ácido Úrico/orina , Xantina , Xantinas/orina

RESUMEN

An ion-paired high-performance liquid chromatographic method was developed for the determination of erythrocyte aldehyde dehydrogenase activity. This method does not require pretreatment to remove hemoglobin from the hemolysate before enzyme reaction is initiated. In addition, the advantage with this high-performance liquid chromatographic method is that erythrocyte aldehyde dehydrogenase activity can be measured using whole blood without either separation or washing of erythrocytes. Therefore, it can be easily used in the determination of erythrocyte aldehyde dehydrogenase activity, which is an indication of excess alcohol consumption.

Asunto(s)

Aldehído Deshidrogenasa/sangre , Eritrocitos/enzimología , Acetaldehído/sangre , Adulto , Anciano , Consumo de Bebidas Alcohólicas/sangre , Cromatografía Líquida de Alta Presión/métodos , Etanol/sangre , Femenino , Humanos , Masculino , Persona de Mediana Edad , NAD/sangre , Valores de Referencia

RESUMEN

Aldehyde oxidase was purified about 120-fold from rat liver cytosol by sequential column chromatography using diethylaminoethyl (DEAE) cellulose, Benzamidine-Sepharose 6B and gel filtration. The purified enzyme was shown as a single band with M(r) of 2.7 x 10(5) on polyacrylamide gel electrophoresis (PAGE) and M(r) of 1.35 x 10(5) on sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Using this purified enzyme, in vitro conversion of allopurinol, pyrazinamide and pyrazinoic acid was investigated. Allopurinol and pyrazinamide were oxidized to oxypurinol and 5-hydroxy-pyrazinamide, respectively, while pyrazinoic acid, the microsomal deamidation product of pyrazinamide, was not oxidized to 5-hydroxypyrazinoic acid. The apparent Km value of the enzyme for pyrazinamide was 160 microM and that for allopurinol was 1.1 mM. On PAGE, allopurinol- or pyrazinamide-stained band was coincident with Coomassie Brilliant Blue R 250-stained band, respectively. These results suggest that aldehyde oxidase may play a role in the oxidation of allopurinol to oxypurinol and that of pyrazinamide to 5-hydroxypyrazinamide with xanthine dehydrogenase which can oxidize both allopurinol and pyrazinamide in vivo. The aldehyde oxidase may also play a major role in the oxidation of allopurinol and pyrazinamide in the subgroup of xanthinuria patients (xanthine oxidase deficiency) who can oxidize both allopurinol and pyrazinamide.

Asunto(s)

Aldehído Oxidorreductasas/metabolismo , Alopurinol/metabolismo , Hígado/enzimología , Pirazinamida/metabolismo , Aldehído Oxidasa , Aldehído Oxidorreductasas/antagonistas & inhibidores , Aldehído Oxidorreductasas/aislamiento & purificación , Animales , Benzamidinas/farmacología , Técnicas In Vitro , Masculino , Oxidación-Reducción , Oxipurinol/metabolismo , Pirazinamida/análogos & derivados , Ratas , Ratas Wistar , Triazinas/farmacología

RESUMEN

Xanthine oxidase was purified 1600-fold from human liver cytosol. The purified enzyme was shown as a single band of 300 kDa on polyacrylamide gel electrophoresis and 150 kDa on SDS-PAGE. Using this purified enzyme, polyclonal antibody against xanthine oxidase was raised in a rabbit. On Ouchterlony's double immunodiffusion method, the raised antibody and the human liver cytosol made a precipitation line stained by activity stain and protein stain, respectively. With the raised anti-xanthine oxidase sera, the immunohistochemical localization of xanthine oxidase in human tissues was examined. Immunostaining of frozen hepatic tissue section showed that the cytoplasm of hepatocytes and endothelial lining cells were stained. In a number of other tissues, the xanthine oxidase antigen was detected only in the endothelial lining cells from heart, kidney, brain, aorta, lung and mesentery, except for the duodenal mucosa cells. A possible role for xanthine oxidase in the endothelial cells from various human tissues in the pathogenesis of reperfusion injury was suggested.

Asunto(s)

Hígado/enzimología , Xantina Oxidasa/aislamiento & purificación , Especificidad de Anticuerpos , Endotelio Vascular/enzimología , Humanos , Inmunohistoquímica , Mucosa Intestinal/enzimología , Hígado/inmunología , Xantina Oxidasa/análisis