Effect of weight loss on inflammation in patients with mild obstructive sleep apnea.

BACKGROUND AND AIMS: Inflammation may be one mediating mechanism for cardiovascular diseases in obstructive sleep apnea (OSA). However, little is known about subclinical inflammation or the effect of lifestyle intervention on inflammation in early stages of OSA. The aim of this substudy of an existing randomized controlled trial, with post hoc analyses, was to determine the impact of lifestyle changes aimed at weight reduction on inflammatory biomarkers in overweight patients with mild OSA. METHODS AND RESULTS: Patients were randomized to supervised intensive lifestyle intervention group (N=28) or to control group (N=31), which received routine lifestyle advices. Circulating concentrations of pro- and anti-inflammatory mediators were measured before and after the 1-year intervention. The concentrations of two pro-inflammatory mediators, high-sensitivity C-reactive protein (hsCRP) and interleukin (IL)-6, decreased significantly in both groups. Although the changes in inflammatory biomarkers favored the supervised lifestyle intervention, the only significant reduction observed between the groups was for the anti-inflammatory IL-1 receptor antagonist (IL-1RA). The change in hsCRP was associated with apnea-hypopnea index, and improving night-time oxygen saturation was related to tumor necrosis factor alpha. IL-1RA and IL-6 were associated with insulin metabolism. CONCLUSION: Weight loss resulted in reductions in concentrations of some pro- and anti-inflammatory mediators in overweight patients with mild OSA, overall favoring the supervised lifestyle intervention. These findings suggest that more intensive treatment of obesity in OSA patients might be well-justified.

Cystatin C, NT-proBNP, and inflammatory markers in acute heart failure: insights into the cardiorenal syndrome.

BACKGROUND: Inflammation is thought to be a mediator in the pathophysiology of the cardiorenal syndrome. We evaluated the interactions between kidney function, cardiac stress, and various inflammatory cytokines in patients with acute heart failure (AHF). The effect on 1-year mortality was also assessed. METHODS AND RESULTS: Plasma levels of cystatin C, NT-proBNP, and inflammatory cytokines (interleukin [IL]-6, tumor necrosis factor-α [TNF-α], IL-10) were measured in consecutive patients (n = 465) hospitalized for AHF. After adjustment for demographic characteristics and comorbidities, TNF-α had the strongest relation with renal function (ß = 0.39, P < 0.0001). Elevated TNF-α levels were seen in patients with high cystatin C, irrespective of NT-proBNP. Levels of IL-6 (ß = 0.26, P < 0.0001) and IL-10 (ß = 0.15, P < 0.01), but not TNF-α, were associated with NT-proBNP. Moreover, the most elevated levels of IL-6 were seen in patients with combined high NT-proBNP and high cystatin C. Cox regression analysis found IL-6 above median to be independently predictive of mortality (hazard ratio 1.9; 95% CI 1.2-2.9, P = 0.003). TNF-α was not significantly associated with prognosis in the overall population after adjustment for multiple covariates, but improved risk stratification in the subgroup with low cystatin C and NT-proBNP. CONCLUSION: Levels of TNF-α in AHF are related to kidney function, but not to NT-proBNP. IL-6 seems to be more associated with cardiac stress. Patients with severe dual organ dysfunction have the highest levels of IL-6 and TNF-α. Different relations of inflammatory cytokines to renal function and cardiac stress need to be considered when evaluating heart--kidney interactions.

Effects of strength and endurance training on metabolic risk factors in healthy 40-65-year-old men.

This study compared 21 weeks of combined high-intensity strength and endurance training with endurance or strength training only on metabolic risk factors in 40-65-year-old men. The healthy men (n=63) were randomized into endurance (E), strength (S), combined strength and endurance training (SE) and control (C) groups. S and E trained two times a week and SE 2+2 times a week. Systolic (SBP) and diastolic blood pressure decreased significantly both in E (-6+/-8 and -4+/-6 mmHg) and in S (-9+/-8 and -5+/-7 mmHg), but not in SE or C (P=0.003 for the difference in the changes of SBP between the groups). The changes in serum glucose and insulin during an oral glucose tolerance test did not differ between the groups. Only E decreased serum fasting insulin levels (-17+/-27%, P=0.013). Minor changes were observed in blood lipids and lipoproteins in all groups. Both endurance and strength training can modestly improve metabolic health even in relatively lean older men with normal glucose tolerance. Combined strength and endurance training did not produce complementary benefits on metabolic risk factors. Combined training is effective in improving body composition and cardiorespiratory and muscular fitness, however, which is likely to decrease the risk of future metabolic and cardiovascular disease.

Polymorphism of the interleukin 1 receptor antagonist (IL1Ra) gene and placental abruption.

Candidate genes with a possible involvement in placental abruption are mainly those related to thrombophilia and preeclampsia. Some reports have shown by placental histologic investigation that increased risk of placental abruption is associated with prolonged inflammation. The polymorphic allele A2 in the gene coding for interleukin 1 receptor antagonist (IL1Ra) has been associated in various diseases of autoimmune or inflammatory nature. In obstetrics, previous research data has linked altered IL1Ra protein production with placental pathology and some severe pregnancy complications. In this study, we have determined whether IL1Ra gene polymorphism is associated also with an increased risk of placental abruption. The study involved 116 women with placental abruption and 112 healthy control pregnant women who were genotyped for polymorphism of the IL1Ra gene. The genotype and allele frequencies were assessed between the two groups and also compared with those in the general population. The frequency of the A2 allele was 28.0% among cases and 33.0% in controls (p=0.29), both similar to that in the general population (28.9%). In addition, the genotype distribution of IL1Ra polymorphisms was similar in both groups. Interestingly, there were a relatively higher number of cases with allele A3 (n=4; 1.7%) compared with the controls (0.4%) and the general population (1.0%) but the difference was not statistically significant. We conclude that there is no significant difference in IL1Ra polymorphisms between patients with and without placental abruption.

Elevated serum erythropoietin concentration is associated with coordinated changes in red blood cell and reticulocyte indices of pregnant women at term.

The aim of this study was to evaluate the relationship between the erythropoietin (EPO) concentration and both the advanced cellular indices reflecting the haemoglobin contents of red blood cells and reticulocytes and the serum markers of iron status. The study population comprised pregnant women at term (n = 210). The serum EPO, transferrin receptor (TfR), ferritin, the percentages of hypochromic red blood cells (%HYPOm) or reticulocytes (%HYPOr) and the cellular haemoglobin in mature red blood cells (CHm) or in reticulocytes (CHr) were measured in maternal blood before delivery. The EPO concentration was elevated above the reference limit (>31.5 mIU/mL) in 16 % of all pregnant women, and appeared to correlate in a linear fashion, especially with %HYPOm (r = 0.52, p<0.001), %HYPOr (r = 0.57, p<0.001) and CHr (r = -0.45, p<0.001). The significant (p<0.05) predictors of EPO in the multivariate stepwise regression analysis were %HYPOr, Hb, %HYPOm and MCV. In general, the lower the cellular haemoglobin content, the higher the overall maternal EPO production. In conclusion, elevated %HYPOm and %HYPOr reflecting iron-deficient erythropoiesis are associated with an increase in EPO concentration in maternal blood. This could be explained by subclinical iron deficiency being accompanied by a compensatory EPO response.

Imprecision of cardiac marker analyses among laboratories on the basis of external quality assurance results: Finnish experience.

The purpose of this report was to evaluate the reproducibility and harmonization of cardiac marker tests and to describe the current situation concerning quality of assays for cardiac markers on the basis of the results of the external quality control schemes (EQAS) of Labquality Ltd., Helsinki, Finland in the period 2002 to 2005. Finnish EQAS surveys obtained for proficiency samples at low marker concentration indicated that the overall coefficient of variation (CV) between laboratories for CK-MBmass and troponin I exceeded 10 %, while for cardiac troponin T the CV was 8.6 %. Intra-laboratory reproducibility was investigated in a single laboratory using concomitant testing in the same EDTA plasma samples to establish cut-off limits for one CK-MBmass and three troponin assays. The 10 % imprecision limit obtained from the concomitant testing in the same samples for CK-MBmass was (by Elecsys) 8.5 microg/L, for cardiac troponin T (by Elecsys) 0.023 microg/L and for cardiac troponin I (by AxSYM) and by Immulite 2000) 0.85 microg/L and 0.63 microg/L. At present, it is recommended that laboratories determine the concentration at which the 10 % imprecision for a specific cardiac marker assay is reached, because the assays generally do not reach that imprecision at the level of the 99th percentile value, usually taken as decisional level. However, common efforts of scientific societies and professional diagnostic industry associations internationally are needed if consensus is to be reached on standardization of immunoassays for cardiac markers and uniform results obtained among laboratories.

Best use of the recommended IFCC reference method, material and values in HbA1C analyses.

The results of Finnish HbA(1C) surveys (Labquality Ltd.) during the past 10 years have undergone continuous improvement with smaller overall coefficients of variation for the HbA(1C) mean values of all methods (from 7.5 to 5.4% for normal and from 8.9 to 4.7% for diabetic samples). Most of the HbA(1C) methods are certified for traceability to the Diabetes Control and Complication Trial (DCCT) designated comparison method, which originally was a high-performance liquid chromatography (HPLC) method (Bio-Rex 70, Bio-Rad) but is no longer in routine use. It was therefore important that the International Federation of Clinical Chemistry and Laboratory Medicine (IFCC) had prepared both reference preparations and method for the determination of HbA(1C). However, the very demanding reference method is not realistic for use in clinical laboratories. According to the present study, the mean HbA(1C) values of the Labquality Ltd. showed significant correlations to the HbA(1C) values of The European Reference Laboratory for Glycohemoglobin (r = 0.999) and to the values using the IFCC method (r = 0.999). The reference values of the IFCC method (mainly those of the manufacturer) range from 2.85 to 3.81%, being significantly lower than the present DCCT values (4.0-6.1%). Since it may take some time before consumers are ready to accept the new IFCC reference values for general use, we propose that the IFCC reference materials and method should be used for calibration of the present methods to the well-known DCCT levels.

High amount of visceral fat mass is associated with multiple metabolic changes in offspring of type 2 diabetic patients.

OBJECTIVE: To investigate the relative contribution of total body fat mass (TFM) and intra-abdominal fat mass (IAFM) to metabolic consequences of obesity in offspring of type 2 diabetic parents. DESIGN: Cross-sectional study of 129 nondiabetic offspring of diabetic parents (59 men, 70 women, age 35.7 +/- 6.3 y, body mass index 26.2 +/- 4.6 kg/m2). Study subjects were grouped according to TFM (assessed with bioelectrical impedance) and IAFM (assessed with CT). Insulin sensitivity was assessed with the euglycemic hyperinsulinemic clamp, insulin secretion with the intravenous glucose tolerance test and energy expenditure with indirect calorimetry. Furthermore, C-reactive protein (CRP) and adiponectin levels were measured. RESULTS: Insulin resistance, low rates of oxidative and nonoxidative glucose disposal, high rates of lipid oxidation and reduced energy expenditure during hyperinsulinemia were associated with high IAFM, independently of TFM. Adiponectin level was reduced and CRP level increased in subjects with high IAFM. CONCLUSIONS: The metabolic changes relating to obesity are largely attributable to high IAFM, and are present even in normal weight subjects with high IAFM.

Biomarkers of alcohol consumption in patients classified according to the degree of liver disease severity.

OBJECTIVE: In the search for optimal biomarkers of excessive drinking, only a few studies have been conducted to compare the relationships between ethanol consumption, liver status, and various laboratory markers of ethanol-induced diseases. MATERIAL AND METHODS: Concentrations of carbohydrate-deficient transferrin (%CDT and CDTect methods), serum sialic acid (SA), gamma-glutamyl transferase (gamma-GT), aspartate aminotransferase (ASAT), mean corpuscular volume (MCV), and a marker of fibrogenesis (PIIINP) were studied in 102 alcoholics with (n=59) or without (n=43) alcoholic liver disease. Controls were 34 healthy volunteers who were either social drinkers or abstainers. RESULTS: Although concentrations of all markers were significantly higher in the alcoholic patients than in the healthy controls, their diagnostic characteristics showed a considerable degree of variation. The %CDT, SA, and MCV showed the strongest correlations with the amount of recent alcohol intake. The presence of liver pathology notably influenced the results of CDTect, GT, ASAT, and PIIINP. In ROC analyses, the highest rates of diagnostic accuracy for detecting hazardous drinking were reached with GT (0.94), CDT (0.86), and SA (0.85), followed by MCV (0.79) and ASAT (0.77). Upon abstinence, the estimated times for normalization varied between 10 days (CDTect) and 25 days (GT). CONCLUSIONS: Our data suggest distinct differences in the clinical characteristics of biological markers of ethanol consumption. While the overall accuracy of CDT and GT appear to be highest in the detection of problem drinking, serum SA and PIIINP measurements are of further value when the effects of liver pathology and ethanol drinking need to be differentiated.

M385T polymorphism in the factor V gene, but not Leiden mutation, is associated with placental abruption in Finnish women.

This study determines whether genetic variability in the gene encoding factor V contributes to differences in susceptibility to placental abruption. Allele and genotype frequencies of three single nucleotide polymorphisms (SNPs) in the factor V gene leading to nonsynonymous changes (M385T in exon 8, and R485K and R506Q [Leiden mutation] in exon 10) were studied in 116 Caucasian women with placental abruption and 112 healthy controls. Single-point analysis was expanded to haplotype analysis and haplotype frequencies were estimated using an expectation-maximisation (EM) algorithm. Comparison of single-point allele and genotype distributions of SNPs in exon 8 and exon 10 of the factor V gene revealed statistically significant differences in M385T allele (P = 0.021) and genotype ( P = 0.013) frequencies between the patients and the control subjects. The C allele of SNP M385T was significantly less frequent among the patients (7%) vs. the control subjects (13%), at an odds ratio of 0.48 (95% CI 0.25-0.91). Allele and genotype differences between the patients and control subjects as regards R485K and Leiden mutation were not significant. In haplotype estimation analysis, there was a significantly lower frequency of haplotype T-R-R encoding the T385-R485-R506 variant in the group with placental abruption vs. the control group (P = 0.038) at an odds ratio of 0.519 (95% CI 0.272-0.987). We conclude that T385 is less frequent among the patient group than in the control group. The M385T variant in the factor V gene other than the Leiden mutation may play a role in disease susceptibility.

C-reactive protein and the development of the metabolic syndrome and diabetes in middle-aged men.

AIMS/HYPOTHESIS: Low-grade inflammation has been implicated in the development of Type 2 diabetes and cardiovascular disease, but its role in the pathogenesis of the metabolic syndrome is unclear. We investigated the association between C-reactive protein (CRP) levels and the development of the metabolic syndrome and diabetes in men. METHODS: Serum CRP concentrations and factors related to insulin resistance were determined in middle-aged Finnish men who participated in a population-based cohort study and were free of diabetes at baseline. RESULTS: At the 11-year follow-up, 143 of 680 men had developed the metabolic syndrome as defined by the National Cholesterol Education Program (NCEP) and 103 of 598 men had developed the metabolic syndrome as defined by the World Health Organization (WHO). Our analyses excluded men with the metabolic syndrome by the respective definition at baseline. In all, 78 of 762 men developed diabetes over the same period. Men with CRP concentrations > or =3 mg/l had a several-fold higher age-adjusted risk of developing the metabolic syndrome (NCEP definition: odds ratio [OR]=3.2, 95% CI 1.9-5.5; WHO definition: OR=3.4, 95% CI 2.0-6.1) or diabetes (OR=4.1, 95% CI 2.1-8.0) than men whose CRP levels were <1.0 mg/l. Even after further adjustment for potentially confounding lifestyle factors and factors related to insulin resistance, the risk of diabetes (OR=2.3, 95% CI 1.0-5.1) was still increased in men with CRP concentrations > or =3 mg/l, but the association with the metabolic syndrome was no longer significant. CONCLUSIONS/INTERPRETATION: Low-grade inflammation may increase the risk of the metabolic syndrome and diabetes in middle-aged men, but some of the risk is mediated through obesity and factors related to insulin resistance.

In patients who have stainable iron in the bone marrow an elevated plasma transferrin receptor value may reflect functional iron deficiency.

Elevated transferrin receptor (TfR) concentration may be either because of iron deficiency or an increased rate of erythropoiesis. We have studied the relationship between elevated TfR and advanced RBC and reticulocyte indices in an unselected population of hospitalized patients. The iron status in 95 consecutive hospitalized patients was assessed using bone marrow aspirate examination and analysis of the RBC and reticulocyte indices was performed using the Advia 120 haematology system. Of the 95 patients, a total of 17 had no stainable iron in the bone marrow and most of them also had an elevated TfR. Of the 78 patients with stainable iron stores, 15 also had an elevated TfR concentration (> or =2.4 mg/l). Six of them also had an elevated %HYPOm (> or =3.4%), and therefore were regarded as having functional iron deficiency. We evaluated the possible causes of elevated TfR concentrations in patients having stainable iron in the bone marrow, and this study suggests that functional iron deficiency explains a considerable proportion of these cases.

Changes in sex hormone-binding globulin and testosterone during weight loss and weight maintenance in abdominally obese men with the metabolic syndrome.

BACKGROUND: Mild hypoandrogenism in men, usually defined by low levels of testosterone, is a peculiar feature of abdominal obesity that independently predicts the development of insulin resistance and diabetes mellitus. Little is known about the short- and long-term effects of weight loss on sex steroids in abdominally obese men, however. OBJECTIVES: We assessed the effect of rapid weight loss and sustained weight maintenance on the plasma concentrations of testosterone and other sex hormones in 58 abdominally obese men (age, 46.3 +/- 7.5 years; body mass index, 36.1 +/- 3.8 kg/m(2); waist girth, 121 +/- 10 cm) with the metabolic syndrome. RESULTS: The men lost on average 16.3 +/- 4.5 kg during a 9-week very low-calorie diet (VLCD) and maintained 14.3 +/- 9.1 kg weight loss after a 12-month maintenance period (vs. baseline, p < 0.001). Sex hormone-binding globulin (SHBG) increased from 27.6 +/- 11.9 to 48.1 +/- 23.5 nmol/l during the VLCD but decreased to 32.6 +/- 12.9 nmol/l during weight maintenance, which was still higher than at baseline (p < 0.001). Free testosterone (fT) increased from 185 +/- 66 to 208 +/- 70 pmol/l (p = 0.002) during the VLCD and remained high after 1 year of weight maintenance (212 +/- 84 pmol/l, p = 0.002). Total testosterone levels followed a pattern intermediate between fT and SHBG. Plasma estradiol and dehydroepiandrosterone sulphate concentrations changed only transiently or not at all. CONCLUSIONS: Rapid weight loss with successful weight maintenance in abdominally obese men with the metabolic syndrome brings about a sustained increase in fT levels. The dramatic increase in SHBG attenuated initially during weight maintenance but remained elevated. These findings may be important with regard to prevention of progressive metabolic decompensation and cardiovascular disease associated with obesity and the metabolic syndrome.

Association of single nucleotide polymorphisms of the bile salt export pump gene with intrahepatic cholestasis of pregnancy.

BACKGROUND: We determined whether genetic variability in the gene encoding the bile salt export pump (BSEP) contributes to individual differences in susceptibility to the development of intrahepatic cholestasis of pregnancy (ICP). METHODS: The study involved 57 affected and 115 healthy control pregnant women who were genotyped for two single nucleotide polymorphisms (SNPs) in the BSEP gene. Chi-square analysis was used to assess genotype and allele frequency differences between the cholestatic and control groups. In addition, single locus analysis was expanded to pair of loci haplotype analysis to examine the estimated haplotype frequencies of the two SNPs, of unknown phase, among the cholestatic and control groups. Estimated haplotype frequencies were assessed using the maximum-likelihood method, employing an expectation-maximization (EM) algorithm. RESULTS: The genotype and allele frequency distribution of the two intragenic SNPs in the ICP and control groups revealed significant evidence of association with the exon 28 SNP (P=0.04 and P=0.02, respectively). In addition, a borderline allele association was noted with the intron 19 SNP (P=0.08). Although the overall distribution of estimated haplotypes of intron 19 and exon 28 SNPs did not differ between the ICP and control groups, the most common haplotype, A-G, was significantly overrepresented in the ICP group (P=0.02), at an odds ratio of 1.73 (95% CI: 1.08-2.74). CONCLUSIONS: The use of two intragenic SNPs in both single locus and haplotype analyses of association suggests that the BSEP gene is a susceptibility gene in intrahepatic cholestasis of pregnancy.

High oxidized LDL and elevated plasma homocysteine contribute to the early reduction of myocardial flow reserve in healthy adults.

BACKGROUND: Impairment of coronary blood flow reserve has been shown to be an early manifestation of atherosclerosis and coronary artery disease (CAD). We studied more closely the contribution of various risk factors on early deterioration of coronary function. MATERIALS AND METHODS: Fifty-one young, apparently healthy adults, with normal or mildly elevated serum cholesterol levels but without other major risk factors for CAD, such as diabetes or hypertension, underwent positron emission tomography (PET) studies. Coronary flow reserve (CFR) was measured using O15-water. In addition to the classical risk factors, the role of several new risk indicators, such as low-density lipoprotein (LDL) oxidation, infection (Chlamydia pneumoniae antibodies), and inflammation parameters (adhesion molecules, ICAM, VCAM, selectin, and C-reactive protein), homocysteine and body iron stores were investigated. RESULTS: Elevated lipid and lipoprotein levels were not associated with reduced coronary reactivity. However, high autoantibody titers against oxidized LDL (oxLDL) were associated with 21% lower CFR than low oxLDL (P < 0.05). Furthermore, high homocysteine levels predicted low CFR (P < 0.05). The other measured parameters, Chlamydia pneumoniae antibody levels, C-reactive protein and adhesion molecule concentrations did not associate with myocardial blood flow. In a stepwise regression model, oxLDL (P = 0.03), homocysteine (P = 0.04) and triglycerides (P = 0.018) were significant predictors of CFR. CONCLUSIONS: The present study suggests an important role for oxidized LDL and plasma homocysteine on early impairment of coronary reactivity in young adults.

Lack of association between C-850T polymorphism of the gene encoding tumor necrosis factor-alpha and polycystic ovary syndrome.

In the present study, we determined whether genetic variability in the gene encoding tumor necrosis factor-alpha (TNF-alpha) contributes to individual differences in susceptibility to the development of polycystic ovary syndrome (PCOS). The study involved 87 Caucasian Finnish women with PCOS and 115 healthy control women who were genotyped for the C-850T polymorphism in the TNF-alpha gene promoter. Analysis by chi(2) was used to assess genotype and allele frequency differences between PCOS women and controls. A similar genotype distribution for the C-850T polymorphism was observed in the two groups, with the frequency of the variant T allele being 8.6% in the PCOS group and 9.6% in the control group (p = 0.862). Accordingly, the profile of genotype frequencies was similar in the groups. The observed profiles of allele and genotype frequencies confirm an equilibrium state between C-850T polymorphism and PCOS and suggest that polymorphism of the TNF-alpha gene is unlikely to contribute to the risk of PCOS.

Autocrine production and synergistic growth-promoting activity of interleukin-6 and oncostatin M in a new human myeloma cell line TU-1.

Human myeloma cell lines are difficult to establish, and they usually originate from patients with extramedullary disease. We describe a new human myeloma cell line, TU-1, which was established from the bone marrow of a patient without extramedullary myeloma. The myeloma cells were initially maintained in a conditioned medium derived from another well-known myeloma cell line U-266. This conditioned medium contained interleukin-6 (IL-6) and oncostatin M (OSM), and possibly other unknown growth factors as well. In 3 months the TU-1 cell line proliferated autonomously and secreted IL-6 and OSM with a synergistic growth response. As we have previously shown the cell line acquired a p53 mutation in vitro, which may be an important factor causing autonomous proliferation. In patients with multiple myeloma OSM is frequently found in the serum and OSM has been associated with serum IL-6 and progressive disease. Our study demonstrates the close relationship of OSM and IL-6 also in vitro.

Serum and plasma as alternative sample types in analysis of cardiac markers in the clinical routine.

There is an increasing demand for the results of cardiac markers (troponin I or T, creatine kinase MB mass and myoglobin) to be made available promptly after sample-taking. In order to shorten the turnaround time, the possibility of using EDTA- or heparin-plasma instead of serum was investigated. The study population comprised 391 patients with acute chest pain. Four different instruments and systems routinely used in Finland giving quantitative results were studied for the assays of creatine kinase isoenzyme MB mass, myoglobin, and troponin I or troponin T. In addition to serum samples, heparin-plasma seems to be useful for all three assays using the Access and Immulite systems, while EDTA-plasma seems to be useful for all three assays with the Access and Elecsys systems. For the AxSYM assays, serum samples seem to be the best alternative. In conclusion, it is possible to use a single EDTA- or heparin-plasma sample for Access, Elecsys and Immulite analysers, and thereby to shorten the turnaround time. In this way the quantitative analyses from plasma can be performed 30 min after taking the sample.