Miloš Tichý

Cytokines and adhesion molecules in the course of acute myocardial infarction

The plasma levels of interleukin 1 beta (IL 1beta), interleukin 6 (IL 6), interleukin 8 (IL 8), tumor necrosis factor alpha (TNF-alpha), E-selectin, ICAM 1 and C-reactive protein (CRP) have been studied in 24 patients with acute myocardial infarction in the course of 96 h. The plasma IL 1beta and IL 6 levels were continually elevated during the 96 h study period (the peak of plasma IL 1beta level was 22.2 pg/ml, S.D. 8.6, P < 0.001, normal values of IL 1beta are less than 10 pg/ml, the mean peak plasma concentration of IL 6 was 184.9 pg/ml, S.D. 134.7, vs. normal values of 15.57 pg/ml, S.D. 2.4, P < 0.001). The mean plasma IL 8 level was increased for the duration of the study, the mean plasma IL 8 level was 103.0 pg/ml, S.D. 23.4 (normal value was below 30 pg/l, S.D. 8.0) P < 0.001. The plasma TNF-alpha level was elevated throughout the time of observation without any significant peak. The mean plasma TNF-alpha concentration was 46.8 pg/ml, S.D. 2.13, vs. normal value 4.35 pg/ml, S.D. 1.23, P < 0.001. The plasma E-selectin level reached the mean level of 145.1 ng/ml, S.D. 75.4, vs. normal value 29.1-63.4 ng/ml, P < 0.001 at an interval of 15-42 h after the onset of the symptoms. The plasma ICAM 1 level showed only a slight significant increase during the first 36 h. The plasma CRP concentration increased later than IL 6, and reached a peak at 42 h after the onset of the symptoms (69.2 mg/l, S.D. 29.9, vs. 1.2 mg/l, S.D. 4.7, P < 0.0001). We conclude that cytokines and adhesion molecules can play an important role in the mechanisms of tissue injury in the process of ischemia and reperfusion.

High-dose chemotherapy followed by autologous stem cell transplantation changes prognosis of IgD multiple myeloma

Immunoglobulin D (IgD) multiple myeloma (MM) is a rare plasma cell disorder constituting less than 2% of all MM cases. Survival of patients with IgD MM is generally shorter than that of patients with other types of monoclonal (M-) protein. We have retrospectively analyzed patients with IgD MM participating in clinical trials of the Czech Myeloma Group. Twenty-six IgD MM patients treated between 1996 and 2006 were identified, 14 (54%) men and 12 (46%) women. The median age was 61 years (range: 37–79 years). Ten of 26 patients (39%) were treated with first-line high-dose chemotherapy (HDCT) using melphalan 200 mg/m2 followed by autologous stem cell transplantation (ASCT). Thirteen of 26 patients (50%) received conventional chemotherapy (CHT), mostly melphalan and prednisone or a vincristine/doxorubicin/dexamethasone (VAD) regimen. Treatment responses were evaluable for 23 of 26 (89%) patients. All HDCT patients had treatment responses, including seven patients (70%) with complete responses and three patients (30%) with partial responses. The median progression-free survival was 18 months for HDCT patients and 20 months for CHT patients. The median overall survival (OS) for all patients was 34 months. The median OS for the HDCT group has not yet been reached (70% of the patients are still alive). In contrast, the median OS for CHT patients was only 16 months. The difference in OS between the two groups was statistically significant (P=0.005). In conclusion, the overall response rate for patients with IgD MM aged 65 years or less treated with HDCT and ASCT is similar to that seen in other MM types.

Use of the biochip microarray system in detection of myocardial injury caused by radiofrequency catheter ablation.

Abstract Background: In a prospective study, we measured plasma markers of myocardial damage induced by radiofrequency catheter ablation (RFA) with the protein biochip microarray system. Methods: A total of 32 consecutive patients undergoing RFA for atrioventricular nodal re-entry tachycardia (AVNRT), right atrial flutter (AFL) and atrial fibrillation (AF) were included in the study. Cardiac troponin I (cTnI), creatine kinase isoenzyme MB (CK-MB), heart-type fatty acid binding protein (hFABP) and glycogen phosphorylase BB (GPBB) were measured using biochip array technology at baseline and 24 h after the procedure. Results: Values for all markers increased 24 h after RFA (cTnI: 0.92±0.49 μg/L vs. 0.33±0.06 μg/L, p<0.001; CK-MB: 3.79±2.04 μg/L vs. 1.85±0.55 μg/L, p<0.001; hFABP: 2.82±0.95 μg/L vs. 2.00±0.95 μg/L, p<0.001; GPBB: 9.07±5.83 μg/L vs. 4.70±2.50 μg/L, p<0.001). The correlations between plasma marker levels and RFA time were cTnI: r=0.63, p<0.01; CK-MB: r=0.75, p<0.01; hFABP: r=0.55, p<0.05, GPBB: r=0.51, p<0.05; the correlation between RFA time and number of RF applications was significant (r=0.81, p<0.001). Patients with RFA due to AF or flutter had elevated cTnI, CK-MB and hFABP levels compared to patients with AVNRT (cTnI: 1.14± 0.49 μg/L vs. 0.59±0.25 μg/L, p<0.05; CK-MB: 4.46± 2.07 μg/L vs. 2.81±1.54 μg/L, p<0.05; hFABP: 3.21± 0.98 μg/L vs. 2.25±0.54 μg/L, p<0.01). Conclusions: Myocardial injury induced by RFA can be detected by cTnI, CK-MB, hFABP and GPBB. Plasma cTnI, CK-MB and hFABP levels significantly increased in patients with AFL and AF compared to patients with AVNRT. The increase of cTnI, CK-MB and GPBB levels correlates with the total duration of RFA. Clin Chem Lab Med 2008;46:1726–8.

Leucine and protein metabolism in rats with chronic renal insufficiency

The aim of this study was to evaluate the effect of chronic uremia induced by 5/6 nephrectomy (5/6NX) on changes in protein and branched-chain amino acid (BCAA; valine, leucine and isoleucine) metabolism. The control group consisted of sham operated rats. Twenty eight weeks after surgery the parameters of protein and amino acid metabolism were evaluated using a primed constant intravenous infusion of L-[1-(14)C]leucine. A drop in BCAA levels and a significant increase in urea, creatinine and cholesterol were observed in plasma of all 5/6NX rats. However, severe uremia with acidosis developed only in one third of rats with 5/6NX. In 5/6NX rats with acidosis significant increases in proteolysis, leucine oxidation, leucine oxidized fraction, and leucine clearance were observed in comparison with the control group and rats with 5/6NX without acidosis. In addition, in 5/6NX rats with acidosis a significant decrease in valine concentration in gastrocnemius muscle was found. We conclude that marked activation of proteolysis occurs in severe chronic renal failure and is probably caused by metabolic changes related to acidosis development.

PLASMA INTERLEUKIN-6 LEVEL IS ASSOCIATED WITH NT-PROBNP LEVEL AND PREDICTS SHORT- AND LONG TERM MORTALITY IN PATIENTS WITH ACUTE HEART FAILURE

OBJECTIVES Interleukin 6 plays an important role in chronic heart failure (HF), but little is known about its involvement in acute decompensated heart failure (ADHF). The aim of our study is to evaluate the prognostic role of interleukin 6 (IL-6) in the patients with ADHF. METHODS Plasma levels of interleukin IL-6, N-terminal pro brain natriuretic peptide levels, and clinical covariates were measured in 92 patients with ADHF. Survival was followed up to 12 months, and prognostic factors were evaluated. RESULTS Elevated plasma IL-6 levels were increased in nonsurvivors and were associated with 1-year mortality (p < 0.01). Plasma IL-6 levels were associated with plasma NT-proBNP levels. In multivariate analysis, increased plasma IL-6 and NT-proBNP levels remained strong independent predictors of 1-year mortality. CONCLUSIONS Plasma IL-6 levels provide important prognostic information in the patients with ADHF. Measurement combining plasma IL-6 and NT-proBNP should serve as a powerful prognostic tool of multimarker strategy in patients with acute decompensated heart failure.

Two-dimensional gel electrophoresis of four serum samples from patients with IgD myeloma

Sera containing IgD paraprotein present problems in identifying patients with monoclonal gammopathies because only a small or even no spike may be present on standard serum protein electrophoresis. We have detected heavy chains of IgD monoclonal protein by means of high resolution two-dimensional gel electrophoresis. Besides clearly identifying delta heavy chains in maps of serum proteins, we also found size and charge heterogeneity of monoclonal immunoglobulins. The results demonstrate the usefulness of two-dimensional gel electrophoresis in the analysis of selected cases of immunoglobulin malignancies.

Interlaboratory study of free monoclonal immunoglobulin light chain quantification

Abstract Background: Quantification of monoclonal immunoglobulin free light chains (FLCs) in serum is used increasingly in clinical practice for the diagnosis, prognostic assessment, and treatment monitoring of monoclonal gammopathies. It is used as an adjunct to standard serum protein electrophoresis and immunofixation. However, methods for FLC quantification need further standardization and validation. Methods: The Czech Myeloma Group and the Czech Society of Clinical Biochemistry have initiated an interlaboratory study where six laboratories collaborating with the primary myeloma treatment centres measured FLC concentrations in 12 serum samples from patients with monoclonal gammopathies. Results: Repeatability of the measurements in five laboratories was calculated based on differences between the results of duplicate measurements. We found that repeatability depended more on the laboratory than on the device used for measurement. Conclusions: The study revealed several weak points in the methodology, including the need for a uniform sample dilution procedure. Interlaboratory reproducibility was comparable with values achieved in the NEQAS programme. Because the κ/λ ratio cannot be measured with high precision, κ and λ FLC concentrations should be used where possible. Due to its impact on the clinical management of patients with gammopathy, FLC quantification needs to become a part of the regular quality control cycle in myeloma centres.

Plasma glycogen phosphorylase BB is associated with pulmonary artery wedge pressure and left ventricle mass index in patients with hypertrophic cardiomyopathy.

Radek Pudil*, Martina Vašatová, Juraj Lenčo, Miloš Tichý, Vı́t Řeháček, Alena Fučı́ková, Jan M. Horáček, Jan Vojáček, Miloslav Pleskot, Jiřı́ Stulı́k and Vladimı́r Palička 1 Faculty of Medicine Hradec Kralove, First Department of Medicine, Charles University Prague, Hradec Kralove, Czech Republic 2 Faculty of Medicine Hradec Kralove, Institute of Clinical Biochemistry and Diagnostics, Charles University Prague, Hradec Kralove, Czech Republic 3 Faculty of Military Health Sciences, Department of Molecular Pathology, University of Defence, Hradec Kralove, Czech Republic

The properties of IgM-L pyroglobulin

Abstract The finding of IgM-L paraprotein with pyroprecipitability was made in the serum of a 65-year-old healthy man. The serum was tested by electrophoresis, immunoelectrophoresis, quantitative radial immunodiffusion, gel filtration and sedimentation analysis. It was demonstrated that the heat precipitability is bound on intact IgM-L paraprotein.

Transient IgM-lambda paraprotein in a 15-month-old child.

Abstract IgM-lambda paraprotein was repeatedly demonstrated in the serum of a 15-month-old girl suffering from an acute inflammatory disease; it disappeared within 4 months.