RESUMO
BACKGROUND: This study aimed to assess the role of intraoperative enteroscopy (IOE) in determining surgical treatment. METHODS: The IOE procedure was performed for 30 patients with Crohn's disease. The degree of stricture and the presence of active ulcer were examined. Preoperative diagnoses and intraoperative findings obtained by inspection and palpation were noted and compared with the IOE findings. RESULTS: Of the 78 intestinal strictures observed by IOE (42%), 33 were not found by preoperative examination. Of the 45 strictures confirmed by IOE to be severe (<15 mm in diameter), 8 were judged to be mild (15-25 mm in diameter) or were not even identified by intraoperative inspection and palpation. Active ulcer was found at 12 of 33 mild strictures, and all 12 strictures were surgically corrected. Of 11 severe strictures detected by IOE at previous surgical sites, 9 were found preoperatively, and 4 were judged to be mild on the basis of inspection and palpation. Stricture was found at the ileocecal valve by IOE in seven patients, but was not diagnosed preoperatively in two of these patients. CONCLUSION: Intraoperative enteroscopy provides useful information regarding the status of the lumen in patients with Crohn's disease.
Assuntos
Doença de Crohn/diagnóstico , Endoscopia Gastrointestinal/métodos , Mucosa Intestinal/patologia , Laparotomia/métodos , Monitorização Intraoperatória/métodos , Adulto , Estudos de Coortes , Doença de Crohn/cirurgia , Tomada de Decisões , Feminino , Humanos , Intestino Delgado/patologia , Intestino Delgado/cirurgia , Masculino , Pessoa de Meia-Idade , Complicações Pós-Operatórias/prevenção & controle , Cuidados Pré-Operatórios/métodos , Prognóstico , Estudos Retrospectivos , Medição de Risco , Sensibilidade e Especificidade , Índice de Gravidade de Doença , Resultado do TratamentoRESUMO
An accurate assay method of 25-hydroxyvitamin D3 24-hydroxylase (24-hydroxylase) was established. Kidney mitochondria prepared from vitamin D-replete rats were treated with polyoxyethylenesorbitan monolaurate. The solubilized suspension was ultracentrifuged at 100,000g for 60 minutes and an aliquot of the supernatant was incubated under the saturating concentrations of substrate NADPH and the mitochondrial-type electron transferring proteins, adrenodoxin and NADPH-adrenodoxin reductase. Products were analyzed by high-performance liquid chromatography (HPLC) monitoring effluents at a wavelength of 265 nm. The maximal velocity of the enzyme in vitamin D-replete rats was 400 pmol/minute per mg of protein, which was considerably higher than those reported by previous authors who used intact kidney mitochondria as the enzyme source. In applying the new assay method, an interesting property was found; Michaelis constant of 24-hydroxylase for 25-hydroxyvitamin D3 [25(OH)D3] was 0.6 microM, which was 35-fold lower than that for 1alpha,25-dihydroxyvitamin D3 [1alpha,25(OH)2D3] which was 20.9 microM. This fact indicates that affinity of the enzyme to 25(OH)D3 is 35-fold higher than that to 1alpha,25(OH)2D3. These data suggest that 25(OH)D3 is the preferred substrate to 1alpha,25(OH)2D3.
Assuntos
25-Hidroxivitamina D3 1-alfa-Hidroxilase/metabolismo , Calcifediol/metabolismo , Calcitriol/metabolismo , 25-Hidroxivitamina D3 1-alfa-Hidroxilase/análise , Adrenodoxina/metabolismo , Animais , Cromatografia Líquida de Alta Pressão , Ferredoxina-NADP Redutase/metabolismo , Concentração de Íons de Hidrogênio , Rim/enzimologia , Rim/metabolismo , Cinética , Masculino , Mitocôndrias/enzimologia , Mitocôndrias/metabolismo , Polissorbatos/farmacologia , Ratos , Ratos Sprague-Dawley , TemperaturaRESUMO
We purified extensively 25-hydroxyvitamin D3 1alpha-hydroxylase (calcidiol, NADPH: oxygen oxidoreductase (1-hydroxylating), EC 1.14.13.13) from kidney mitochondria of rachitic rats and disclosed its peculiar properties as a P450. The final preparation was identified as a 55 kDa protein having an intense absorption at 417 nm characteristic of P450. The specific activity was 4.8 nmol/min/mg of protein indicating a 350-fold purification. Specific content of P450 was 1.1 nmol/mg of protein and turnover number was 4.4 min-1.
Assuntos
25-Hidroxivitamina D3 1-alfa-Hidroxilase/isolamento & purificação , Rim/enzimologia , Mitocôndrias/enzimologia , 25-Hidroxivitamina D3 1-alfa-Hidroxilase/química , 25-Hidroxivitamina D3 1-alfa-Hidroxilase/metabolismo , Animais , Cromatografia Líquida/métodos , Sistema Enzimático do Citocromo P-450/análise , Estabilidade Enzimática , Masculino , Peso Molecular , Ratos , Ratos Sprague-Dawley , RaquitismoRESUMO
An assay method for 25-hydroxyvitamin D3 1alpha-hydroxylase [calcidiol, NADPH: oxygen oxidoreductase (1-hydroxylating), EC 1.14. 13.13] in rat kidney is described. The mitochondrial and nuclear fraction was solubilized effectively with 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate(Chaps). By subsequent ultracentrifugation of the solubilized suspension the effect of inhibitory factor(s) in mammals was removed. The enzyme was then assayed by the reconstitution method using saturated amounts of adrenodoxin and NADPH-adrenodoxin reductase. Products were analyzed by HPLC, monitoring absorbance at 265 nm. The enzyme activity depended on not only pH of the medium but also the kind of buffers. N,N-Bis(2-hydroxyethyl)glycine was the best buffer. At 30 degrees C, the reaction velocity was linear at least up to 10 min, by which time enough amounts of the product needed for analysis were formed. The enzyme activity was linear to a protein concentration up to 0.8 mg of protein/ml. Under the best assay conditions established, the maximal velocity of enzyme in the rachitic rat was 12.9 pmol of product/min/mg of protein, which was 30- to 1000-fold higher than those reported by other authors with the enzyme of rachitic rat. Michaelis constant was 1.8 microM. Specific activity with the enzyme of normal rat was 0.25 pmol of product/min/mg.