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N W Tietz

Publications and source records attributed to N W Tietz.

At least 19 recordsLinked to original sources

Interassay calibration as a major contribution to the comparability of results in clinical enzymology.

OBJECTIVE: Factors contributing to the applicability of interassay calibration of methods measuring enzyme catalytic activities are described. Also discussed are the properties essential for such a material. Similarity of specificity for the methods to be calibrated as well as commutability between the material(s) intended to be used as calibrator are the main criteria to be satisfied. RESULT: Several examples demonstrated that interassay calibration is feasible but a multi-enzyme calibrator with a wide commutability for the most popular methods remains to be developed. This is the project of the IFCC Working Group on Calibrators in Clinical Enzymology (WG-CCE). Several experimental data are also presented that indicate that the temperature at which the reaction is carried out is not a limiting factor in the implementation of interassay calibration in clinical enzymology.

Calibration↗

Validation of an enzyme calibrator--an IFCC guideline. International Federation of Clinical Chemistry.

OBJECTIVES: The objective of this guideline is to improve standardization in clinical enzymology in order to improve intermethod comparability of patients' results. DESIGN AND METHODS: The reference system, combination of the reference method and the reference material, is used to produce a reference value for a given catalytic activity. Sets of methods are formed of methods exhibiting the same analytical specificity. Materials intended to be used as enzyme calibrators are experimentally checked for their commutability. RESULTS: The transfer of accuracy from the reference value to patients' results is dependent on methods (analytical specificity) and on materials (experimentally assessed commutability). The feasibility of this approach was demonstrated with materials of high level for several enzymes and for each of them for several routine methods. CONCLUSION: Expected advantages of this approach in clinical enzymology are presented.

Calibration↗

Support of the diagnosis of pancreatitis by enzyme tests--old problems, new techniques.

Computerized tomography and ultrasound are helpful in the diagnosis of acute pancreatitis. The procedures, however, are not always available and so laboratory tests continue to play an important role. Serum amylase measurement is the most widely used test, despite its lack of sensitivity (75-92%) and specificity (20-60%). A cut-off point of 3-6 times the upper reference limit increases the specificity greatly, but at the expense of sensitivity. A method using chloronitrophenyl-maltotrioside as substrate has recently been rejected by the IFCC. A method using ethylidene-protected 4-nitrophenyl-maltoheptaoside and a new alpha-glucosidase has emerged as the method of choice. Amylase isoenzyme determinations have higher specificity than total amylase measurements. Serum lipase methods are more sensitive and specific, but methodological problems persist. Cationic trypsin-1 measurements yield high sensitivity but low specificity. Elastase-1 is claimed to correlate best with the clinical symptoms. Reasons for the widely differing reports on sensitivity and specificity of tests for pancreatitis are discussed.

Clinical Enzyme Tests↗

Possible concentration-dependent suppression of immune response by verapamil.

Calcium channel blockers inhibit in vitro the lymphocyte response to mitogens and the generation of cytotoxic T-cell and natural killer cell activity. We report on a patient taking verapamil hydrochloride who experienced repeated, prolonged viral infections. His lymphocyte response to phytohemagglutinin, pokeweed mitogen, and tetanus antigen were suppressed; the mononuclear response to soluble tetanus was markedly diminished; and there was no response to influenza vaccination. On lowering the verapamil dose, or changing therapy to clonidine hydrochloride, all the above-mentioned functions returned to normal and a response to influenza vaccination was observed. We speculate that the prolonged viral infection observed in this patient may have been caused by suppression of the immune response by verapamil and we encourage further boad-based studies.

Aged↗

Accuracy in clinical chemistry--does anybody care?

The role of clinical chemistry in the diagnostic process has been enhanced by the evolution of better instrumentation and analytical techniques. The quality of some laboratory tests, however, has not kept pace with these advances. I present three examples--serum iron, serum lipase, and nonisotopic immunoassays--assays where some currently used methodologies are highly flawed. Causes for the less than optimal performance of some clinical laboratories are discussed.

Chemistry, Clinical↗

When is a serum iron really a serum iron? The status of serum iron measurements.

Although serum iron determinations play an important role in the diagnostic process, the accuracy of routine methods is suspect. The poor performance of currently used methods is well documented in the quarterly College of American Pathologists survey reports. Compared with the 1990 method of the International Committee for Standardization in Haematology (ICSH), the 1978 ICSH Reference Method shows a +3.4% bias and the standard method of the National Committee for Clinical Laboratory Standards shows a -17.8% bias for iron results. All routine methods checked--DuPont aca, Abbott TDx, Kodak Ektachem (except one lot), BM/Hitachi 717, and Synermed--show a negative bias over the entire analytical range, a significant negative intercept, and extremely poor correlation with the 1990 ICSH method for iron values < 750 micrograms/L. Individual discrepancies of several hundred percent were observed. Imprecision of the methods is not the reason for these discrepancies. We question whether most routine methods measuring low iron concentrations provide results sufficiently reliable for confirmation of iron deficiency.

Autoanalysis↗

Lipase in serum--the elusive enzyme: an overview.

Lipase is a glycoprotein with 420-449 amino acid residues and a M(r) of 46,000-56,000 for pancreatic lipase and 32,000-39,000 for serum lipase. Lipase is present in the pancreas, intestines, and a variety of other tissues. The concentration gradient between pancreatic tissue and serum lipase is approximately 20,000-fold. Serine, as part of an Asp-His-Ser triad, is the nucleophilic residue essential for catalysis. Lipase differs from other esterases by the presence of a hydrophobic recognition site. The optimal pH is between 7.5 and 10.0, depending on the reaction condition; the pI for the various forms of the enzyme has been reported as 5.80 and 5.85; 6.4, 6.8, and 7.0; and 7.4 for a purified fraction. Several authors report the presence of two molecular forms in the pancreas and three electrophoretic bands with lipolytic activity. In normal serum two bands have been observed; in pancreatitis as many as four bands have been seen. Lipolytic activity may not always be due to lipase. Assays specific for lipase require a triglyceride as substrate as well as the presence of colipase (a water-soluble and heat-stable protein, essential for lipase action), a secondary bile salt, and Ca2+. The clinical sensitivity of all modern assays is high because of selection of a low decision limit; the clinical specificity varies greatly but can be improved by increasing the cutoff point. Lipase determinations in pancreatitis are superior to amylase determinations. The reasons for the great variability of reports regarding the clinical utility of lipase are discussed, and the clinical utility of lipase determinations is summarized.

Chemical Phenomena↗

Are clinical laboratory proficiency tests as good as they can be?

Present proficiency test services that use the peer group mean and statistically derived ranges of acceptability are not serving us optimally and are even counterproductive in some respects. We recommend that the target value be determined by a widely accepted reference method and that acceptable ranges be based on criteria related to clinical need. This approach was adopted several years ago in Germany and has already eliminated the use of several unsatisfactory analytical methods. Because the transition would probably take many years, we propose an interim solution to allow instrument manufacturers and laboratorians to adapt to these changes. The current peer group means and acceptable ranges should be supplemented by reference method values and acceptable ranges, based on clinical need, so that manufacturers and laboratorians can judge their performance against these new criteria and make the necessary adjustment in instrumentation and methodology. These processes should be paralleled by efforts to produce proficiency test materials that will not exhibit the matrix problems of present-day preparations.

Chemistry, Clinical↗

Laboratory values in fit aging individuals--sexagenarians through centenarians.

We determined approximately 15,000 laboratory values in 236 individuals between the ages of 60 and 90 y, 22 individuals between 90 and 99 y, and 69 individuals greater than or equal to 100 y, and compared these with values in young adults. We tested 47 different analytes in the 60-90-y group and 93 analytes in the greater than or equal to 90-y group. Na, K, Cl, and CO2 values were either identical or showed minimal change with age; pH decreased slightly. Differences in Ca values were only minor, but ionized Ca increased slightly. Phosphate decreased in men, but changed only minimally in women; parathyroid hormone increased with age. Increases with age were also observed for glucose, insulin, and C-peptide. Among the enzymes, alkaline phosphatase increased in women, but in men only greater than 90 y; gamma-glutamyltransferase increased in both sexes. Creatine kinase (CK) decreased slightly in individuals greater than 70 y and markedly in those greater than 90 y of age, whereas CK-MB decreased markedly greater than 70 y, reaching the detection limit in individuals greater than 90 y. Lactate dehydrogenase isoenzyme 5 decreased slightly with age. Urea nitrogen increased gradually with age, but creatinine increased only in individuals greater than or equal to 90 y. The increase in urea is not paralleled by a loss of protein in urine, suggesting that the possible cause of azotemia may not always be renal pathology. Urate increased in women but not in men. Liver function, as measured by total bilirubin and liver enzymes, was exceedingly well maintained. Concentrations of most proteins show little change, except for slight decreases in prealbumin, albumin, and transferrin, proteins used as an index of nutritional status. IgA values increased, IgG ranges were wider, IgM and IgD decreased, and the range for IgE was narrower than in young adults. Cholesterol, high-density lipoprotein cholesterol, and triglyceride values increased with age, but decreased in individuals greater than or equal to 90 y. Among the trace elements, magnesium changed little, zinc and lead decreased, and copper values increased with age. Total triiodothyronine and thyroxine decreased, with concomitant increases in thyroid-stimulating hormone. More individuals had increased microsomal antibodies and thyroglobulin titers in the aging population than in the young. In men, the free, percent free, bioactive, and total testosterone values decreased, but luteinizing hormone (LH) and follicle-stimulating hormone (FSH) values increased. In women, estrone and estradiol values decreased, with concomitant increases in LH and FSH. Androstenedione and progesterone decreased in both sexes.(ABSTRACT TRUNCATED AT 400 WORDS)

Adult↗

Beta-hCG vs intact hCG assays in the detection of trophoblastic disease.

We describe a case of choriocarcinoma in which increased concentrations of beta-choriogonadotropin subunits in serum indicated recurrence of metastasis long before the tumor could be confirmed by visualization techniques, while intact choriogonadotropin remained normal (less than 5 int. units/L). Therefore, detection or monitoring of choriocarcinoma requires the assay of free beta-hCG subunit or an assay that measures both the free beta-subunit and intact choriogonadotropin.

Adult↗

Lipase activity measured in serum by a continuous-monitoring pH-Stat technique--an update.

Using recent knowledge regarding the roles of colipase, bile acids, Ca2+, and emulsifiers, we optimized a previously published pH-Stat method for lipase (EC 3.1.1.3) activity measurements. The recommended assay conditions are: olive oil/triolein, 100 mL/L; sodium glycocholate, 35 mmol/L; Ca2+, 8.5 mmol/L; and colipase, 6.0 mg/L. The sample volume is 0.10 mL, the reaction pH 9.0, the temperature 30 degrees C, and the concentration of titrant 15 mmol/L. Hydroxypropyl methylcellulose, 20 g/L, replaces acacia as emulsifier to avoid inhibition by excess Ca2+. The standard curve is linear to greater than 4566 U/L. The reference interval with olive oil as substrate is 30-235 U/L. Lipase activities with triolein substrate are 9.9% greater than with olive oil. Interference by pancreatic carboxylesterase (EC 3.1.1.1) activity is inhibited by incubating the sample with diisopropylfluorophosphate. Results correlate well with those by the optimized SingleVial method of Boehringer Mannheim Diagnostics (r = 0.997) and the immunochemical assay of Beckman Instruments, Inc. (r = 0.995). Correlation with the aca method (E.I. DuPont de Nemours & Company) is less satisfactory (r = 0.892), probably owing to lack of colipase in the latter method.

Adult↗

Multicenter evaluation of a specific pancreatic isoamylase assay based on a double monoclonal-antibody technique.

Eleven evaluators from nine laboratories in five countries evaluated a new immunoinhibition method for pancreatic isoamylase determination that is as simple to perform as that for total amylase. The precision at low and intermediate activity concentrations was superior, and at high concentrations it equalled that of the wheat-germ inhibitor method. The test was linear to approximately 2000 U/L, depending on the instrumentation used. The percentage salivary isoamylase activities remaining in specimens after reaction with two monoclonal antibodies ranged from 2 to 4.4%. Comparative studies showed good correlation with the wheat-germ inhibitor (r greater than 0.978) and electrophoresis methods (r = 0.920). Hemolysis, lipemia, and bilirubinemia have no effect on results. Interlaboratory studies demonstrated excellent transferability of the method, if instruments are calibrated with the same calibrator. Reference intervals for pancreatic isoamylase are 13 to 64 U/L (25 degrees C), 13 to 83 U/L (30 degrees C), and 17 to 115 U/L (37 degrees C). A clinical evaluation of patients with acute pancreatitis showed that pancreatic isoamylase has a greater clinical sensitivity than total amylase.

Acute Disease↗

Turbidimetric measurement of lipase activity--problems and some solutions.

We optimized a commercial turbidimetric method for lipase (EC 3.1.1.3) activity (Boehringer Mannheim Diagnostics) and overcame some of its deficiencies. Increasing the bile salt concentration to 35 mmol/L and the colipase concentration to 6 mg/L and using a continuous recording of the reaction-rate curve greatly improved the reaction kinetics, eliminated false results from increases in absorbance, reduced the lag phase, and increased the analytical sensitivity and accuracy. Differentiation of pancreatitis from nonpancreatitis sera by adding NaCl, 140 mmol/L, to the assay mixture to observe the degree of enzyme activation has important limitations. Sera from patients with pancreatitis and only slight or modest increases in lipase behave like sera from healthy individuals or from patients with nonpancreatic disease. The assay shows no interference by lipoprotein lipase and carboxyl esterase. Results compare well by this optimized method and by an optimized "pH-Stat" titrimetric method.

Adult↗