E. Knoll

Urinary excretion of N-acetyl-β-d-glucosaminidase and alanine aminopeptidase in patients receiving amikacin or cis-platinum

Urinary excretion of alanine aminopeptidase (EC 3.4.11.2) and N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30) was determined for 25-70 days in five patients receiving cis-platinum and for 8-53 days in six patients receiving amikacin. This study was performed to investigate if the excretion of urinary enzymes represents a sensitive parameter for the early detection of toxic kidney damage. The determination of N-acetyl-beta-D-glucosaminidase was carried out by the method of Knoll et al. [13]. The procedure of Mondorf et al. [14] for the estimation of alanine aminopeptidase activity was adapted to the Gemsaec Fast-Analyzer. In both patient groups an increase in the excretion of the two enzyme activities could be demonstrated. In patients receiving amikacin, the excretion of alanine aminopeptidase was always higher than that of N-acetyl-beta-D-glucosaminidase, whereas in three patients receiving cis-platinum it was the opposite. In two cis-platinum patients the excretion of both enzymes was of the same size. The changes during amikacin therapy seem to be reversible, whereas in four cis-platinum patients these changes seemed to be partly irreversible. Serum creatinine concentration was less sensitive than the urinary enzyme excretion for detection of kidney damage.

Blood Loss from Laboratory Tests

BACKGROUND Laboratory tests can be an important source of blood loss in hospitals, especially for newborns and patients in intensive care. The aim of this study was to quantify blood loss for laboratory diagnostic tests in a large number of patients in a teaching hospital. METHODS We estimated blood loss by multiplying the number and volumes of sampling tubes collected from 2654 adult inpatients. We compared the number of tests per patient for all inpatients and intensive care unit patients during the first period and again in the same time period 1 year later when cumulative blood-loss volumes were being reported to physicians and educational information had been given to decrease blood loss from laboratory tests. RESULTS For 95% of the patients, blood loss during hospitalization was <196 mL. The largest proportion of the blood samples was used for clinical chemical tests (median, 45%), followed by hematologic (median, 26%) and coagulation (median, 17%) tests. In the surgical and cardiovascular surgical intensive care units, however, blood gas analyses accounted for 19-34% (medians) of the use. For 5% of the patients, all undergoing intensive care, blood loss was >200 mL and for 0.7% was >600 mL during their hospital stay. Such high blood losses were observed in patients with long-term ventilation, coagulation disorders, and repeated surgery. The largest median blood loss was in patients undergoing cardiovascular surgery (median, 201 mL). The mean number of tests was 44 per inpatient before cumulative blood loss was being reported and 46 when it was being reported. CONCLUSIONS Blood loss from laboratory diagnostic testing is not likely to pose a problem for most hospitalized patients. Blood loss is greater in intensive care patients and after cardiovascular surgical procedures. Reporting of the cumulative individual blood loss did not decrease blood loss for laboratory testing.

Plasma free and conjugated catecholamines in diagnosis and localisation of pheochromocytoma

In six patients urinary excretion of vanillylmandelic acid and catecholamines (CA) could establish the diagnosis of pheochromocytoma. Free norepinephrine (NE) in plasma was within the normal range in two patients and plasma free epinephrine (E) was only marginally elevated in one of them. The degree of CA conjugation was not altered and scattered as in controls and was therefore not complementary to the usual determination of plasma free CA. The intermittent nature of CA secretion by the tumour could be demonstrated by multiple blood samplings during a 48-h study period in two patients, e.g. normal plasma values might be associated with pheochromocytoma if measurements are made during a trough. Thus a single peripheral CA determination cannot be of discriminative value in the diagnosis of pheochromocytoma unless it shows marked elevation. Ten patients subjected to intracardial measurements and five patients suspected of having a pheochromocytoma underwent venous catheterisation to determine their free and conjugated plasma CA. In controls CA values near the orifices of adrenal veins differed enormously and partly overlapped with corresponding levels of patients with pheochromocytoma. In one patient with surgically proven left adrenal tumour highest concentrations of CA were measured in the vena cava superior. These high CA concentrations, caused by paroxysmal release of CA by the tumour arouse suspicion of an additional, ectopic tumour. Because venous catheterisation cannot be relied on implicitly we propose computed tomographic scanning as a first step in localisation of a pheochromocytoma.

Evidence for a diagnostic window in fourth generation assays for HIV.

BACKGROUND The routine HIV screening essentially depends on the detection of HIV specific antibodies. However, HIV p24 antigen can be detected in individuals with recent HIV infection about 2-18 days prior to seroconversion. New fourth generation HIV screening assays combine the detection of anti-HIV antibodies with the simultaneous detection of HIV p24 antigen. This may result in a reduction of the diagnostic window after primary infection. OBJECTIVES The performance of two novel fourth generation assays in routine diagnostic was evaluated. STUDY DESIGN We compared two third generation, two fourth generation and one antigen HIV assays in a case with acute primary HIV infection. RESULTS In our case, the HIV infection was detected 11 days earlier with the fourth generation assays compared to third generation assays. Interestingly, after the initial reactive results the fourth generation assays became negative resulting in a second diagnostic window. During this second diagnostic window neither third nor fourth generation HIV assays were reactive. This second diagnostic window was caused by the absence of HIV specific antibodies and the decline of HIV p24 antigen concentrations below the detection limits of the fourth generation assays. CONCLUSIONS Fourth generation assays markedly improve the diagnosis of recent HIV infections but the possibility of a second diagnostic window must be considered.

Plasma free and conjugated catecholamines in clinical disorders

The data suggest that sulfation of CA may not simply be ascribed to platelets, to the liver, to vascular beds, or to organs along the vena cava including the adrenal glands. The unchanged degree of conjugation in patients with increased spillover of NE and E point to a balance between free and sulfated CA. A normal ratio of free to conjugated NE and E observed in patients receiving high dosage DA infusion further indicates that there is an adequate sulfate supply and no apparent substrate inhibition of the conjugation process. Hypothyroidism may affect the balance of free to conjugated CA in a yet unknown way.

Influence of Food Intake on Concentrations of Plasma Catecholamines and Cortisol

Plasma concentrations of epinephrine, norepinephrine, dopamine and cortisol were determined in four healthy volunteers--resting completely in bed--before and after food intake. Meals were provided at 17.00 h on the first day and at 12.00 h on the second day of the test. Blood samples were drawn every 15 minutes. Epinephrine, norepinephrine, dopamine, and cortisol showed a clear dependence on food intake. After the meals plasma concentrations of the four analytes showed rises, which were more pronounced after lunch at 12.00 h than after dinner at 17.00 h. An increase of epinephrine was observed shortly before meal-times; this could be caused by physiological adaptation to the usual rhythm of food intake and reflex response to expectation of the meal.

Determination of catecholamines in plasma by HPLC and amperometric detection. Comparison with a radioenzymatic method

The determination of norepinephrine and epinephrine in plasma by HPLC with amperometric detection was modified, giving detection limits of 25 ng/l for norepinephrine and epinephrine, respectively, using 1 ml plasma. In order to achieve this sensitivity, it was necessary to minimize the background noise by modification of instrumentation and specimen handling. Particularly important was the extra purification of the reagents, the application of micro-bore HPLC, the enzymatic cleavage of uric acid and temperature control of the amperometric cell and the amplifier. Comparison of the present method with the radioenzymatic determination of catecholamines resulted in coefficients of correlation of r = 0.924 and 0.919 for norepinephrine and epinephrine, resp. (n = 38). The concentrations of the 38 different samples used for the comparison were in the physiological range.

Preparation of antibodies to catecholamines and metabolites — Syntheses of various immunogens and characterization of the resulting antibodies

The synthesis of antigens of the haptens p-tyramine, octopamine, synephrine, dopamine, normetanephrine, beta-(3,4-dichlorophenyl)-ethylamine and 3-fluoro-tyramine is described. The coupling of the haptens to the carrier protein was performed either via the side chain by means of succinic anhydride, or through the phenolic ring via an aminomethyl group which was introduced by a Mannich reaction. Antisera to these haptens were produced in rabbits by immunization with these antigens. The antisera were characterized by determination of titer and specificity (p-tyramine, synephrine, normetanephrine). The possibility is discussed of using antisera to p-hydroxyphenylethylamine for a radioimmunological determination of catecholamines.

Urinary Excretion of Alanine Aminopeptidase and N-Acetyl-β-D-Glucosaminidase During Sequential Combination Chemotherapy

The urinary excretion of alanine aminopeptidase (EC 3.4.11.2) and N-acetyl-beta-D-glucosaminidase (EC 3.2.1.30) was determined in 23 patients with testicular cancer during sequential combination chemotherapy with vinblastine/bleomycin and doxorubicin/cis-platinum. Increases in enzyme excretion were more often noticed during therapy with vinblastine/bleomycin than with doxorubicin/cis-platinum. Moreover, the rises during vinblastine/bleomycin therapy were more pronounced. The enzyme activities varied from the normal range up to the 14-fold of the upper limit of the normal range. With few exceptions, enzyme excretions returned to normal or slightly elevated values before the subsequent course. No renal insufficiency could be detected with the commonly used parameters of renal function such as serum creatinine concentrations and creatinine clearance values, which were determined at irregular intervals.

Vergleichende Untersuchung zur diagnostischen Wertigkeit von Diskelektrophorese der Urinproteine und N-Acetylglucosaminidaseausscheidung zur Erkennung von tubulären Nierenschädigungen bei chronischer Polyarthritis

A method is described for the determination of the enzyme N-acetyl-beta-D-glucosaminidase in gel-filtered urine. The individual reaction parameters were tested, and the reliability of the method was determined. The stability of the enzyme in urine was investigated at different temperatures over a 6 week period. The excretion of N-acetyl-beta-D-glucosaminidase, like the analysis of urinary proteins by disc electrophoresis, is a sensitive parameter for renal damage. The two methods were compared, using 50 random urine samples from patients with chronic polyarthritis; both methods were found to have equal diagnostic value.