Evidence for existence of at least two types of factor-VIII-associated non-B transfusion hepatitis.
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Biomedical subjects
Publications and source records attributed to R J Spooner.
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Eight serum enzyme tests were performed over a three-year period in 1,147 cases of patients with suspected hepatobiliary disease, of whom 580 had identifiable primary disease of the liver or biliary system. Individually, aminotransferase assays did not provide good discrimination among the various categories of hepatobiliary disease, but when expressed as a ratio a useful degree of discrimination was obtained. Isocitrate dehydrogenase, guanase and glutamate dehydrogenase alone were poor discriminants of the various disease categories studied; combination of the latter enzyme with the aminotransferases in various ratios did not achieve worthwhile improvement. Adenosine deaminase was normal in most patients with extrahepatic obstruction and abnormal in most patients with parenchymal hepatic disease, and is potentially a useful test additional to the aminotransferases in routine diagnosis. 5'-Nucleotidase was more sensitive and specific than alkaline phosphatase in diagnosing hepatobiliary disorders. Abnormalities of all these enzymes were encountered in patients who did not have hepatobiliary disease, most frequently among subjects with cancer, diabetes mellitus, and diseases of the respiratory and cardiovascular systems.
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A system for the mechanized calculation and transcription of output from a reaction rate analyser has been evaluated in the performance of 8 enzyme assays, many of which show anomalous kinetic behaviour with regard to "lag" or "accelerated" phases or "blank" reactions. For all of these enzymes, a cycle of three consecutive 18-second readings per assay proved optimal, the result being based upon the final output value unless this showed marked discrepancy with the second value, in which circumstances it is helpful to refer to a strip chart recording of the progress curve. Excellent agreement was obtained with manually-derived results and the system functioned without downtime for more than one year. Marked descrepancies between manual and mechanised results were invariably traced to technician error.
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An improved and optimized method for serum glutathione reductase is described. The reference range for normal subjects is 47-79 IU/1. The method is more sensitive than conventional enzyme tests in the detection of malignant disease. It was not raised more frequently in patients with clinical evidence of metastases than in those clinically free of such metastases, and it did not seem to correlate with prognosis among those patients who failed to survive six months from the time the analysis was first conducted.
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Serum glutathione reductase activity decreases on standing at room temperature. Dilution of the sample slows this decay in sera from patients with a variety of clinical disorders, and reverses the inactivation in sera from patients with liver or biliary tract disease. Prolonged contact with dithioeythritol or acetate buffer, pH 6.4, at 4 degrees C prevents spontaneous inactivation of the native serum and abolishes the effect of dilution. These hitherto unreported phenomena are a source of potential inaccuracy in assays for this enzyme. The inactivation may be the result of conformational folding with "burying" of active sites, or of molecular aggregation brought about by hydrogen bonding or disulfide bond formation.
An assessment of the Gilford Automatic Enzyme Analyser was conducted over a period of one year. The optics of the instrument were satisfactory with regard to accuracy of wavelength selection and linearity of absorbance response. Excellent precision was obtained for both absorbance readings and operation of the dispenser pump. Carry-over within the microflow-cell was low. The method of operation recommended by the manufacturers for enzyme determinations failed to take account of endogenous blank reactions which could lead to significant error. When revised methods utilising a pre-incubation stage and initiation with a single substrate were employed, the results correlated well with those obtained with standard automatic (LKB 8600) and manual (Pye Unicam SP 800) kinetic systems for aspartate and alanine aminotransferase, creatine phosphokinase and alpha-hydroxybutyrate dehydrogenase, and the precision at all activity levels was satisfactory. Acceptable precision could not be obtained over the clinical range for enzyme assays requiring a blank determination on each sample (5'-nucleotidase and adenosine deaminase) and those with very low normal serum activities (isocitrate dehydrogenase and glutamate dehydrogenase). These limitations appeared to be due to relative insensitivity of the transducer response and liability to optical disturbance. This apart, the instrument has many advantages over alternative equipment.
Hyperamylasaemia has long been regarded as pathognomonic of acute pancreatitis. However, recent work has revealed a number of conditions where a gross elevation may be an incidental finding, notably diabetic ketoacidosis. The recent discovery of 'macroamylase', a high molecular weight amylase-protein complex capable of producing hyperamylasaemia with low urine amylase, has further complicated diagnosis and has led to the introduction of the ratio of amylase clearance to creatinine clearance as a diagnostic aid. Serum amylase may be resolved, by most electrophoretic media, into bands which correspond to those obtained when pancreatic homogenates or saliva are electrophoresed. The initial promise of this technique has not been realised at the routine diagnostic level. Duodenal juice amylase has been the classical enzyme used in assessing exocrine pancreatic function and although it is still of value it is being amplified by other enzyme tests.
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