Clinical Chemistry and Laboratory Medicine (CCLM)

Published by De Gruyter

Volume 37 Issue 2

Issue of Clinical Chemistry and Laboratory Medicine (CCLM)

Contents
Journal Overview

June 1, 2005

Evolution or Revolution in Clinical Chemistry

Matthew J. McQueen

Page range: 89-90

June 1, 2005

Low Levels of Plasma Proteins: Malnutrition or Inflammation?

Andrew Myron Johnson

Page range: 91-96

Abstract

Levels of several plasma proteins, including albumin, transferrin, and transthyretin (prealbumin), have been proposed as markers for protein energy malnutrition. However, many other factors, especially inflammatory disease and drug or hormone therapy, affect levels of these proteins. These factors probably account for the majority of low levels of transthyretin. Levels of albumin and other proteins may be helpful in determining increased risk of morbidity and mortality, but better markers are needed for diagnosis of protein energy malnutrition per se .

June 1, 2005

Laboratory Medicine: The Need for a Broader View. The “Multiple Bundle” Model of Clinical Laboratory Function

Marek H. Dominiczak

Page range: 97-100

Abstract

The essence of the nineties in health care, in business, in organizational management and in education, has been change. As always in a changing environment, there will be winners and losers. In the September issue of CCLM, Williamson wrote: “Poor clinical chemistry. It is a field trapped between pressures from increasing electronic automation of assays, simplified technology and reductionism of molecular genetics and the growing pressure of economic accountability and cost cutting. It may not survive” (1). Should we all be on Prozac and wait for the doomsday? Our problems are not unique. Some time ago, traditional cardiology was “trapped” between the advent of new invasive techniques on the one hand and a pressure to increase emphasis on prevention on the other. How did it end? Most of today's cardiologists are invasive cardiologists and many became leaders in cardiovascular prevention in addition to their “traditional” tasks (2). This is a classical example of a paradigm shift. The present article suggests that at least some of our problems may stem from too narrow a view of laboratory medicine that we present to decision makers who allocate funds to laboratory services.

June 1, 2005

Activated Protein C (APC)-Resistance: Automated Detection of the Point Mutation at Position 1691 in the Factor V Gene

Karl Rolf Klingler, Stefan Grote, Guido Holzem, Klaus Wielckens

Page range: 101-107

Abstract

Proteolytic cleavage of factor Va, caused by activated protein C, is an important mechanism in limiting clot formation in normal haemostasis. A single point mutation in the factor V gene has been demonstrated to cause resistance of factor Va to proteolytic cleavage by activated protein C. With an 8-fold increased risk of thrombosis and a 2 to 13% prevalence in the Caucasian population for the heterozygous state of this mutation, knowledge of the patient's genetic disposition is of great importance in conditions such as pregnancy, surgery, use of oral contraceptives and immobilization. Therefore we have developed an automated test for the detection of the factor V mutation. This PCR-based test makes use of the disappearance of an Mnl 1 restriction site if the mutation is present. The assay has been developed for the widely used ES-systems of Boehringer Mannheim. The test discriminates between the heterozygous and the homozygous state. Because of its low costs and easy handling the assay can be used as a screening test and can be performed in routine clinical laboratories.

June 1, 2005

A Rapid and Sensitive Automated Light Scattering Immunoassay for Serum C-Reactive Protein and the Definition of a Reference Range in Healthy Blood Donors

Christopher P. Price, Jacqui Calvin, Susan A. Walker, Andrew Trull, David J. Newman, Eileen G. Gorman

Page range: 109-113

Abstract

The increasing interest in the measurement of serum C-reactive protein in relation to the risk stratification of patients with chest pain has demonstrated the need for more sensitive routine methods of measurement, and an accurate definition of the reference range. We report the determination of a reference range in serum samples from 491 blood donors using a particle enhanced turbidimetric immunoassay that has been modified to offer better imprecision within the reference range. The median values were found to be 2.40 and 2.20 mg/l for males and females, respectively, with 95th percentile range of 1.20–5.20 and 0.40–5.40 mg/l respectively.

June 1, 2005

Photodynamic Effects In Vitro in Fresh GynecologicTumors Analyzed with a Bioluminescence Method

Viola Schlosser, Ossi R. Koechli, Rosanna Cattaneo, Brigitte Jentsch, Urs Haller, Heinrich Walt

Page range: 115-120

Abstract

Photodynamic therapy (PDT) is a promising alternative method for clinical cancer treatment. In the present study, cells from four breast carcinomas, seven ovarian carcinomas of various stages of differentiation, and ascites from a diffuse metastatic tumor were treated by PDT in vitro . Tetra(m-hydroxyphenyl)-chlorin (m-THPC) was used as the photosensitizer. Surviving cell rate was evaluated by the ATP-Cell-Viability-Assay (ATP-CVA), which measures light production as an interaction of intracellular ATP with the luciferin-luciferase complex. The most effective PDT of the tumor cells was achieved at an m-THPC concentration of 0.2 μg/ml following incubation of the cells with photosensitizer for 24 hours. PDT toxicity resulted in a cell survival rate of 1 % to 42 % compared to untreated control cells (survival rate of control = 100 %). The inhibitor concentration IC50 of m-THPC was determined both in the dark (dark toxicity) and in combination with laser irradiation. IC50 was defined as the concentration of photosensitizer which caused 50 % of cell death. The IC50 values were heterogeneous in all tumor specimens examined. IC50 values for dark toxicity were on average 0.14 μg m-THPC/ml for primary ovarian carcinoma, 2.16 μg m-THPC ml for refractory ovarian carcinoma and 0.3 μg m-THPC/ml for breast carcinoma. After PDT, average IC50 value for refractory ovarian carcinoma was 0.04 μg m-THPC/ml, for primary ovarian carcinoma 0.05 μg m-THPC/ml and for breast carcinoma 0.03 μg m-THPC/ml. These data might indicate that clinical PDT of gynecological carcinoma requires individual treatment conditions to achieve optimal results.

June 1, 2005

An ELISA for the H-Subunit of Human Ferritin which Employs a Combination of Rabbit Poly- and Mice Monoclonal Antibodies and an Enzyme Labeled Anti-Mouse-IgG

Boris Vaisman, Paolo Santambrogio, Paolo Arosio, Eitan Fibach, Abraham M. Konijn

Page range: 121-125

Abstract

We describe a sensitive ELISA for measuring the H-type subunit of human ferritin. A high detection sensitivity was attained by the use of antibodies from different species and an enzyme-conjugated secondary antibody. It consisted of a sandwich assay using a solid phase coated with a rabbit polyclonal antibody for human ferritin from term placenta and a soluble monoclonal antibody for human H-ferritin, followed by a secondary anti-mouse immunoglobulin (Ig)G conjugated to β-galactosidase. The assay was calibrated with purified recombinant human H-ferritin from E. coli . The colorigenic chlorophenol red β-D-galactopyranoside and the fluorogenic 4-methyl-umbelliferyl-β-D-galactopyranoside substrates were used with similar outcome. The described method permits the measurement of human H-ferritin at a concentration ranging from 0.1 to 100 μg/l (or 20–20000 pg per 200 μl sample) and is accurate at a concentration as low as 0.3 μg/l. The coefficient of variation of the assay was 6.05–10.3 % and the recovery of H-ferritin added to cell lysates was 105.8 ± 7.52 %. Depending on the H-ferritin content of the cell line tested, only 600 to 60 000 cells of different human cell lines were needed to measure their H-territin content.

June 1, 2005

The Use of Free Cortisol Index for Laboratory Assessment of Pituitary-Adrenal Function

Harry A. Bonte, Ron J. van den Hoven, Gertjan van der Sluijs Veer, István Vermes

Page range: 127-132

Abstract

We developed a time-resolved-fluoro-immunoassay to measure cortisol binding globulin (CBG) in serum. It is a microtitre plate, solid phase, reagent excess, sandwich assay in which the same polyclonal antiserum is used as a source of capture and labeled antibodies. The results of this assay were shown to be reliable and were fully comparable with those obtained by a commercially available kit. As a reflection of the free cortisol concentration we measured cortisol and CBG concentrations in serum and calculated the Free Cortisol Index (FCI) = [cortisol] serum /[CBG] serum · 100. The clinical use of this parameter, as a screening test for disturbances of the pituitary-adrenal axis, was investigated in different groups of subjects: healthy men and women, women using oral contraceptives, pregnant women at term, patients with thyroidal illnesses, patients using anti-epileptic drugs and patients suffering from adrenal diseases. In a number of groups we compared the FCI results with measurements of cortisol in saliva, another parameter used as an estimate of the concentration of free cortisol in blood. Our conclusion is that the FCI, in contrast to a total cortisol measurement alone, can prevent unnecessary further testing.

June 1, 2005

Allele-Specific Amplification for the Diagnosis of Autosomal Recessive Spinal Muscular Atrophy

Claire Ravard-Goulvestre, Catherine Boucly, Bertille Mathieu, Geneviève Van Amerongen, Louis Viollet, Brigitte Estournet, Annie Barois, Philippe de Mazancourt

Page range: 133-135

Abstract

The SMN1 gene is homozygously deleted for at least exon 7, interrupted or converted to a non-functional telomeric copy in most cases of proximal spinal muscular atrophies. The presence of a pseudogene hampers direct detection of the exon 7 deletion. We describe a method for the detection of the of exon 7 deletion, based on the amplification refractory mutation system (ARMS), in a multiplex PCR with fluorescent-labelled primers. The gene and pseudogene amplification products differ in the dye bound and in their size, which allows distinction of both products on electrophoresis. The pseudogene is used as an internal control, and this method gives a clear and specific pattern for the patients. Amplification is achieved with 30 cycles, and specificity is retained up to 40 cycles.

June 1, 2005

Diagnostic Value of Biochemical Markers of Bone Turnover and Postmenopausal Osteoporosis

Necat Yılmaz, Metin Bayram, A. Binnur Erbaǧcı, M. Şahin Kılınçer

Page range: 137-143

Abstract

We studied 77 women divided into postmenopausal osteoporotic and premenopausal and postmenopausal non-osteoporotic groups in order to evaluate bone metabolism and diagnostic value of biochemical markers of bone turnover in postmenopausal osteoporosis. Postmenopausal osteoporotic (n: 40), postmenopausal non-osteoporotic (n: 24) and premenopausal nonosteoporotic (n: 13) groups were defined according to bone mineral density (BMD) scores obtained with dual energy X-ray absorptiometry (DEXA). Urinary deoxypyridinoline (Dpd), pyridinoline (Pyd), serum total alkaline phosphatase (ALP), bone specific alkaline phosphatase (BALP), osteocalcin (BGP), total calcium, phosphorus, and creatinine levels were determined. Urinary Dpd and Pyd levels of postmenopausal osteoporotic group (8.7 and 18.7 μmol/mg creatinine) were significantly higher than postmenopausal control (5.1 and 11.7 μmol/mg creatinine, p<0.0001) and premenopausal control (6.0 and 13.0 μmol/mg creatinine, p<0.0005 and p<0.001) groups. Bone formation markers were not significantly different between groups, although BGP correlated with Dpd and Pyd (r: 0.26 and r: 0.31, p<0.05) in osteoporotic subjects. From receiver operating curve (ROC) analysis Dpd had the best diagnostic value (0.846), followed by Pyd (0.802) in evaluation of osteoporosis, whereas BALP (0.570) and BGP (0.528) were relatively inefficient in the discrimination of postmenopausal osteoporosis. This study suggests that bone resorption markers are more efficient than bone formation markers in the diagnosis of postmenopausal osteoporosis. Urinary Dpd/creatinine ratio has the highest diagnostic value.

June 1, 2005

Isoenzmyes of Class I and II Alcohol Dehydrogenase in Chronic Hepatitis

Lech Chrostek, Maciej Szmitkowski

Page range: 145-147

Abstract

Using class-specific fluorogenic substrates, the activities of class I and II alcohol dehydrogenase (ADH) isoenzymes were determined in the sera of patients with chronic hepatitis. The activity of the total alcohol dehydrogenase and indicator enzymes of liver damage were also investigated. We found a statistically significant increase of class I alcohol dehydrogenase isoenzymes in the total tested group which included those with the viral hepatitis. The (2-fold) increase in the activity of class I isoenzymes was similar to the increase of aminotransferases. The serum activity of class II isoenzymes was unchanged. Here an increase in total enzyme activity was not statistically significant. Class I isoenzymes and total enzyme activity correlated well with aminotransferases. These results demonstrate that serum activity of class I ADH measured with fluorogenic substrates confirms liver cell damage and may be useful in the diagnosis of chronic hepatitis.

June 1, 2005

Screening School Children for Albuminuria, Proteinuria and Occult Blood with Dipsticks

Michael J. Pugia, John A. Lott, Junko Kajima, Takaaki Saambe, Miyuki Sasaki, Kooichi Kuromoto, Reiko Nakamura, Hisae Fusegawa, Yoshihide Ohta

Page range: 149-157

Abstract

Beginning in 1974, the Japanese Ministry of Health Welfare directed the screening of schoolchildren for proteinuria. We studied their procedure and methods in 6197 school children and also evaluated a new urine dipstick that measures albumin concentrations down to about 10 mg/l and creatinine down to about 300 mg/l. We used specimens from adult in- and outpatients to test the accuracy of the dipsticks. Based on the quantitative results, we set as cutoffs < 150 mg/l for protein and < 30 mg/l for albumin as the concentrations representing “low risk.” The quantitative values were assumed to be correct, and the dipstick results were judged accordingly, i.e., a dipstick protein of ≥ “150” mg/l or an albumin of | “30” mg/l indicated increased risk of developing or having a genitourinary disorder. The sensitivity/specificity of the protein dipstick was 95.1%/95.5%, and the same for the albumin dipstick was 83.8%/93.8%. The cut-off for the albumin dipsticks probably should be set somewhat lower to reduce the number of false negatives and increase the sensitivity of the dipstick. When we compared the quantitative albumin to the protein dipsticks with the above cut-offs, we found the sensitivity/specificity to be 79.3%/94.4%, i.e., much like the albumin dipstick results. The many reports on the association of albuminuria and risk of renal disease recommend that screening should be done for albumin rather than protein. Based on the data from the school children, we estimate that a dipstick albumin of “30” mg/l is borderline increased risk, and that a protein dipstick of “150” mg/l is the same. If we call the dipstick “10” mg/l albumin, “30” mg/l albumin and the “150” mg/l protein results “low risk,” then we estimate the prevalence of albuminuria in the school children to be about 2.1% and proteinuria to be about 4.3%. Children with these values should have a quantitative test for albumin and protein. We also tested a dipstick for creatinine and found increasing values with increasing age in both genders; the older boys had significantly higher creatinine values than the older girls and younger boys. For the albumin/creatinine ratio, we found 6028 children with a ratio of < 30 mg/g indicating low risk and 159 children with a ratio of ≥30 mg/g indicating increased risk. The ratio may be more useful owing to the likely reduction of the number of false negatives and false positives. Note: 1 mg/l albumin = 8.85 μmol/l .

June 1, 2005

Evaluation of the First Automated Thyroglobulin Assay

Michael Vogeser, Peter Knesewitsch, Karl Jacob, Dietrich Seidel

Page range: 159-164

Abstract

The aim of this study was to investigate technical and analytical performance of the first automated thyroglobulin (Tg) assay (DPC-Immulite ® ; Diagnostic Products Corporation, Los Angeles, USA). In imprecision studies using several human serum pools ranging from 21 to 58 replicates, a coefficient of variation of 9.0 % was obtained at a mean Tg concentration of 0.84 ng/ml and of 6.1 % at a Tg concentration of 62.1 ng/ml. In a method comparison with a non-automated assay (BRAHMS LUMItest Tg ® , BRAHMS, Berlin, Germany) using 383 sera of 303 patients with thyroid carcinoma, regression analysis according to Passing and Bablock yielded in the following equation: Immulite Tg=1.6 × BRAHMS Tg−0.1 ng/ml (Pearson's r=0.979). Sera obtained from 59 patients with thyroid carcinoma enabled comparative follow-up studies; in all cases qualitative agreement was found with regard to increase or decrease of serum Tg; in eight cases, however, Tg was detected with the Immulite assay but not with the BRAHMS assay. Further follow-up proved the presence of thyroid tissue in these patients. From these and further methodological data (dilution linearity, interference studies, carry-over study, high-dose hook properties, and short report time) it is concluded that the DPC-Immulite Tg assay meets the requirements of routine diagnostic use.

June 1, 2005

Selective Imidazoline Receptor Agonists and Lipid Metabolism

Moses Elisaf, Alekos Tselepis, Kostas Siamopoulos

Page range: 165-165

June 1, 2005

Judgement on Analytical Quality Requirements from Published Clinical Vignette Studies Is Flawed

Callum G. Fraser

Page range: 167-168

June 1, 2005

The Meaning of Good Laboratory Practice (GLP) for the Medical Laboratory

Rainer Haeckel

Page range: 169-169

June 1, 2005

Improved Procedure of Measuring on Radiometer 500 and 600 Series Bloodgas Analysers

Gottfried A. Harff, Maria J. A. van den Bosch

Page range: 171-172

July 27, 2005

Scientific Papers and Presentations by Martha Davis

Jacqueline M. Atkinson

Page range: 173-173

July 27, 2005

New Myocardial Marker Proteins in Acute Myocardial Infarction. Quantitative Aspects by J. A. Kragten

R. J. Spooner

Page range: 174-174

July 27, 2005

Normalwerte by Michael Jakob

C. van Heyningen

Page range: 175-175

July 27, 2005

Meetings

Page range: 176-178

About this journal

Objective
Clinical Chemistry and Laboratory Medicine (CCLM) publishes articles on novel teaching and training methods applicable to laboratory medicine. It is focused on basic and applied research and cutting-edge clinical laboratory medicine. CCLM is one of the leading journals in the field, with an impact factor over 8. All contributions submitted for publication in CCLM are single-blind peer reviewed by at least two experts in the field. CCLM is led by a multi-institutional editorial board. It is issued monthly, both in print and electronically. Letters to the Editor and Congress Abstracts are published online only.

Please submit your manuscript here

Topics

clinical biochemistry
clinical genomics and molecular biology
clinical haematology and coagulation
clinical immunology and autoimmunity
clinical microbiology
drug monitoring and analysis
evaluation of diagnostic biomarkers
disease-oriented topics (cardiovascular disease, cancer diagnostics, diabetes)
new reagents, instrumentation and technologies
new methodologies
reference materials and methods
reference values and decision limits
quality and safety in laboratory medicine
translational laboratory medicine
clinical metrology

Article formats
Research Articles, Reviews, Mini Reviews and Opinion Papers on all aspects of clinical chemistry and laboratory medicine, Guidelines and Recommendations, Point/Counterpoint Articles, Letters to the Editor, Editorials

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