Ustav Laboratorni Diagnostiky
Ustav Laboratorni Diagnostiky
Kelbich P.,R.Ø.S.A. |
Kelbich P.,Krajska zdravotni a.s. |
Kelbich P.,Charles University |
Hejcl A.,Masarykova nemocnice v Usti nad Labem |
And 7 more authors.
Klinicka Biochemie a Metabolismus | Year: 2013
Objective: 1. Evaluate the numbers of cells in the cerebrospinal fluid (CSF), glucose concentrations in the CSF, values of the glucose quotient (Q glu.), lactate concentrations in the CSF and values of the coefficient of energy balance (KEB) as indicators of intensity of the inflammatory process in the CSF in groups of patients without CNS impairment, with slight serous inflammation of non-infectious aetiology in the CNS, with serous inflammation of infectious aetiology in the CNS and of patients with purulent inflammation in the CNS with extracellular bacteria in pathogenesis. 2. Compare the information potential of the used parameters of the glucose energy metabolism in the CSF compartments in our group of the investigated patients, i.e. concentrations of glucose in the CSF, values of the Qglu., concentrations of lactate in the CSF and values of the KEB. Design: Retrospective study. Material and Methods: We examined 133 CSF specimens in patients without CNS impairment, 227 CSF specimens in patients with slight serous inflammation with intrathecal synthesis of immunoglobulins of non-infectious aetiology in the CNS, 208 CSF specimens in patients with serous inflammation of infectious aetiology in the CNS and 140 CSF specimens in patients with purulent inflammation in the CNS with extracellular bacteria in pathogenesis. The objects of our interest were numbers of cells in the CSF, concentrations of glucose in the CSF, values of the Qglu., concentrations of lactate in the CSF and values of the KEB. The D'Agostino Omnibus test, the Kruskal-Wallis test with follow-up post hoc analysis using the Dunn's method, the Spearman correlation coefficient and the multinomial logistic regression analysis were used for statistical analysis of the examined parameters. Results: We did not find any changes in the numbers of cells in the CSF and in energy ratios in the CSF compartment of patients without CNS impairment. We found raised numbers of cells in the CSF and slight alterations of the glucose quotients, lactate concentrations in the CSF and the values of the KEB only in some patients with slight serous inflammations of non-infectious aetiology in the CNS. We observed manifestations of conspicuously increased intensity of inflammation in the numbers of cells in the CSF, lactate concentrations in the CSF and the values of the KEB in patients with serous inflammations of infectious aetiology in the CNS. Very high intensity of purulent inflammation in the CNS of bacterial aetiology was well apparent in all the evaluated parameters. Concerning the relationship, either direct or indirect, between the number of cells in the CSF and the other parameters, we found the highest correlation between the number of cells in the CSF and the values of the KEB (ρ = -0.770), followed by the lactate concentrations in the CSF (ρ = 0.734), the Qglu. (ρ = -0.676) and the glucose concentrations in the CSF (ρ = -0.544). We verified the applicability of the parameters mentioned above for prediction of the intensity of inflammation in the CNS via multinomial logistic regression analysis. The number of cells and the KEB, with 71.9 % and 71.6 % respectively, has the highest prediction potential of the correctly classified patients. They were followed by the lactate concentration in the CSF with 64.7 %, the Qglu. with 58.8 % and the glucose concentration with 54.7 % of the correctly classified patients. Conclusion: Our study supports the applicability of the numbers of cells in the CSF, the glucose concentrations in the CSF, the values of the Qglu., the lactate concentrations in the CSF and the values of the KEB for diagnosing CSF impairment and for monitoring the intensity of inflammation in the CNS. Further, the results enabled determination of the information potential of the energy parameters. The values of the KEB were most suitable for evaluation of the intensity of inflammation in the CNS. Less suitable results were achieved in case of the lactate concentrations in the CSF. Even worse results were observed in case of the values of Qglu. and the least suitable results were observed in case of the glucose concentrations in the CSF. © Ceská lékarská spolecnost Jana Evangelisty Purkyne, Praha 2013.
Dolecek R.,Ustav Laboratorni Diagnostiky |
Crkvenjas-Nemeckova Z.,Ustav Laboratorni Diagnostiky |
Pleva L.,Ustav Laboratorni Diagnostiky |
Tvrdik J.,Ustav Laboratorni Diagnostiky |
Svagera Z.,Ustav Laboratorni Diagnostiky
Osteologicky Bulletin | Year: 2015
Osteoporosis following burns has been repeatedly reported. The main objectives were to monitor 28 various markers in a group of 73 burnt patients (12 females, 61 males) on 7 occasions throughout a year (Days 1, 7, 14, 28, 56, 183 and 365), and to determine which markers could contribute to the development of osteoporosis, with general catabolism in acute phase response (APR) that may be excessive. So far, there are no clear criteria for such excessiveness although it has been repeatedly shown to exist. For six to twelve months, bone resorption markers such as CTX, NTX, PHP, DPD and, partly, acid phosphatase were significantly increased. For several weeks, the levels of catabolic interleukin-6 were increased; IL-10 was slightly increased for only a week after burns were sustained. Males had a decrease in anabolism resulting from a significant decrease in testosterone, dihydrotestosterone and f-testosterone lasting for several weeks. Both males and females had a temporary significant decrease in DHEA-S. Following burns, the levels of ionized calcium (iCa) and total calcium (tCa) in blood are significantly lower, with the decreased lasting for longer in case of iCa. The concentrations of PTH were relatively normal, with exceptions noted. In those who died after sustaining burns, however, the concentrations were often very high. Following burns, the levels of "anabolism" markers gradually rise such as alkaline phosphatase (ALP), osteocalcin, IGF-1 and des-acyl ghrelin. Initially, burn patients typically have high levels of 17β-estradiol; Cortisol levels are close to the upper limit of the normal range, with some extremely high as well as low levels being present. Urinary free Cortisol was high for several weeks. The mean levels of calcidiol, or 25 (OH)D, were lower than normal, and low in many individuals. There was a significant difference in many markers following burns and polytrauma. After polytrauma, PTH was very high, blocking IL-10 was high, both iCa and tCa were less reduced, and 17β-estradiol was lower. There was a high increase in ALP in polytrauma. Apparently, APR is sometimes excessive which might even cause harm to the burnt patient. In treatment, anabolics should be considered; an attempt to reduce the excessive APR should be made (successful administration of the neuroplegic mixture M2 in the past should be therefore remembered); and vitamin D3 should be administered following burns.
Zeman D.,Ustav Laboratorni Diagnostiky |
Kusnierova P.,Ustav Laboratorni Diagnostiky |
Gottwaldova J.,Oddeleni Klinicke Biochemie |
Kloudova A.,Oddeleni Imunologie A Alergologie |
And 5 more authors.
Klinicka Biochemie a Metabolismus | Year: 2012
Objective: Quantitation of albumin and immunoglobulins in CSF and serum serves the purpose of calculation of intrathecal immunoglobulin synthesis. Quotient, i.e., concentration ratio of particular protein in CSF and serum, is assumed to be method-independent value, provided that paired CSF and serum sample is analyzed in parallel using one (CSF) calibration curve. We have observed that CSF (but not serum) IgM concentrations determined by nephelometry on Immage analyzer and by home-made sandwich ELISA were largely discrepant. This has lead us to perform a larger study comparing various methods of IgM determination in CSF and serum. Design: Comparison of various methods used for IgM quantitation in cerebrospinal fluid. Materials and Methods: Results of IgM determination on nephelometers Immage, Immage 800, BN II, and BN ProSpec, as well as by ELISA method, have been compared. Beside CSF and serum values, CSF/serum quotients and results of calculated intrathecal IgM synthesis according to Reiber's formula have been compared. Results have been evaluated by means of Passing-Bablok regression and Bland and Altman plots, using MedCalc software. Results: We have found significant differences between CSF IgM concentrations determined by rate nephelometry on Immage analyzers and end-point nephelometry on BN analyzers or ELISA, the latter two giving significantly lower values. CSF IgM concentrations determined by ELISA were in good agreement with those determined on BN analyzers. Serum IgM concentrations were slightly higher using ELISA than using either nephelometric system. We have found no significant difference in either serum IgM or CSF/serum IgM quotient between serum analysis on CSF versus serum calibration curve. IgM quotient values were significantly higher using Immage analyzers than using either BN analyzers or ELISA method. As a consequence, intrathecal IgM synthesis, based on the Immage data, was calculated in 13 of 71 samples considered negative if data of BN analyzer or ELISA were used; in 8 of these samples, intrathecal IgM synthesis could be considered as clinically significant (intrathecal fraction > 10 %). Conclusions: CSF/serum IgM quotient is far from to be method-independent; in some cases, discrepant conclusion regarding the presence of intrathecal IgM synthesis can be obtained. Until the cause of this discrepancy is found and removed, the presence of intrathecal IgM synthesis should be judged very carefully and in the context of other CSF findings when using the Immage analyzer for measurement. ELISA method is inexpensive, requires very low amount of the sample, and is the only method able to quantify IgM in almost every CSF sample. Its use in routine practice is, however, limited by higher requirements for manual work. Nevertheless, it should be used at least in experimental studies, i.e. for the purpose of the determination of distribution of CSF IgM concentration within a population.
Kacirova I.,Ustav klinicke farmakologie |
Kacirova I.,Ustav laboratorni diagnostiky |
Grundmann M.,Ustav klinicke farmakologie
Kardiologicka Revue | Year: 2015
A key strategy in optimizing aminoglycosides and vancomycin therapy is therapeutic drug monitoring. It is a specific method of clinical pharmacology used to monitor the therapy using measurement of drug serum concentrations followed by interpretation by a clinical pharmacologist/pharmacist and good cooperation with the clinician. Therapeutic drug monitoring helps clinicians to quickly optimize aminoglycosides and vancomycin dosing regimens to maximize the clinical effect, minimize the toxicity of the drugs, decrease mortality and morbidity and reduce costs. Aminoglycosides (amikacin and gentamicin) constitute one of the oldest classes of antimicrobials. Despite their relative toxicity, mainly nephrotoxicity and ototoxicity, aminoglycosides are valuable in current clinical practice. They are bactericidal agents used against aerobic gram-negative infections, and in combination with a cell wall active antimicrobial-based regimen (e. g. b-lactams), also against gram-positive cocci. Aminoglycosides have a concentration-dependent bactericidal effect and a long post-antibiotic effect. There is accumulating evidence to show that large, single, daily doses (or more correctly, extended interval dosing) of aminoglycosides are associated with lower nephroand ototoxicity and comparable, if not superior, clinical outcomes than the same total dose administered in small, multiple doses. A general therapeutic range of aminoglycosides does not exist. Every patient has his/her own optimal target concentration based on the microorganism susceptibility, co-administered antibacterials, immune status and co-administration of other nephro-or ototoxic drugs. Minimum serum vancomycin trough concentrations should always be maintained above 10 mg/L to avoid development of resistance, nevertheless, trough concentrations > 20 mg/L are not recommended because of the risk of nephrotoxicity. For serious gram-positive infections vancomycin trough concentrations of 15-20 mg/L are recommended. In non-complicated infections (urinary tract infections or mild-to-moderate skin and soft tissue infections) trough concentrations of 10-15 mg/L should be sufficient. For continuous infusions of vancomycin target steady-state concentration values of 15-25 mg/L is optimal. We demonstrate some case reports of therapeutic monitoring of aminoglycoside antibiotics and vancomycine from our routine practice. © 2015, Ambit Media a.s. All rights reserved.
Grundmann M.,Ustav klinicke farmakologie |
Kacirova I.,Ustav klinicke farmakologie |
Kacirova I.,Ustav laboratorni diagnostiky
Kardiologicka Revue | Year: 2015
Introduction: Digoxin is a positive inotropic drug frequently prescribed in the treatment of chronic congestive cardiac failure. Recent evidence suggests that a lower therapeutic range of 0.5-0.9 ng/mL is associated with reduced mortality. With complex pharmacokinetic profile and narrow therapeutic index, its use in managing patients with heart failure can present a challenge to clinicians. Aim: To show the advantages of therapeutic drug monitoring for dosing of digoxin. Method: Bayesian analysis was used to predict the long-term serum concentration time profiles of digoxin, using the MW-Pharm 3.30 software. The serum levels of digoxin were determined by MEIA. Results: Three case reports are presented showing a prediction of steady-state digoxin level three days after the start of administration, drug-drug interaction between digoxin and spironolactone/carvedilol and an example of long-term patient non-compliance. Conclusion: The- rapeutic drug monitoring is very useful for prediction of serum levels of digoxin alone and in combination with different interacting drugs. It helps to understand compliance-influencing factors and to improve interventional strategies to increase digoxin compliance. © 2015, Ambit Media a.s. All rights reserved.
Zeman D.,Katedra biomedicinskych oboru |
Zeman D.,Ustav laboratorni diagnostiky |
Kusnierova P.,Katedra biomedicinskych oboru |
Kusnierova P.,Ustav laboratorni diagnostiky
Klinicka Biochemie a Metabolismus | Year: 2016
Objective: Improvement of the procedure used for cerebrospinal fluid (CSF) and serum oligoclonal IgM detection. Design: Methodical study Material and Methods: Isoelectric focusing (IEF) in agarose gel followed by affinity immunoblotting. After choosing the best variant from the protocols tested, 35 paired CSF and serum samples were examined. Results: Improvement of IgM separation has been achieved by using narrower pH gradient (4-8), prefocusing step and cathodic application of samples. Efforts to separate IgM in native CSF or serum samples have been unsuccessful. Satisfactory results have been obtained using sample pretreatment with dithiothreitol, optionally followed by acrylamide pretreatment. Intrathecal synthesis of oligoclonal IgM was demonstrated in 11 out of 35 patients. Samples of symptomatic controls (n=3) or patients with a proved diagnosis of non-inflammatory neurological disease (n=9) were all negative. Oligoclonal IgM result correlated significantly with intrathecal IgM synthesis estimated either by Reiber's (P=0.007) or Auer's formula (P=0.026). Conclusion: The method described in this article enables sensitive and reproducible detection of oligoclonal IgM bands separated by IEF. Further research should address the optimisation of sample pretreatment as well as experiments aimed at separation of IgM in untreated cerebrospinal fluid and serum samples.
Pika T.,Hemato Onkologicka Klinika |
Lochman P.,Oddeleni Klinicke Biochemie |
Kusnierova P.,Ustav Laboratorni Diagnostiky |
Hermanova Z.,Ustav Imunologie |
And 4 more authors.
Klinicka Biochemie a Metabolismus | Year: 2015
Introduction: Waldenström's macroglobulinemia (WM) is a rare malignant B-lymphoproliferative disorder, characterized by bone marrow infiltration by tumor cells of lymphoplasmocytic lymphoma (LPL) with production of IgM monoclonal immunoglobulin (MIg). The newest test for MIg is the HevyLite system, based on the assessment using the pair of specific antibodies against junction epitopes between the domains of heavy and light chain (HLC) of the constant region of immunoglobulin chains. Aim: The aim of our paper was the comparison of detection methods for serum levels of MIg of IgM isotype in patients with newly diagnosed WM using conventional electrophoresis and with the use of the levels of heavy/light chain pairs of the immunoglobulin. Patients and methods: Our cohort consisted of 15 sera of WM patients with IgM kappa isotype. The samples were assessed using conventional gel and capillary electrophoresis. The assessment of MIg using gel electrophoresis was carried out using Sebia Hydrasys system, for capillary electrophoresis we used Sebia MiniCap system. For turbidimetry analysis of HLC pairs we used HevyLite Human Ig sets, IgM kappa kit for the use on the SPAplus with the use of turbidimetry SPAplus. For nephelometric analysis of HLC pairs we used nephelometer BN II and HevyLite IgMκ. Results: Within the comparison of the levels of MIg using gel (5 - 60 g/l) and capillary (7.5 - 62.5 g/l) electrophoresis we found strong correllation (r = 0.937, p < 0.0001) with median difference 13%. Median total serum protein concentration was 95.3 g/l (73.4 - 131.6 g/l). Comparison of MIg detected using gel electrophoresis and IgMκ levels detected using SPAplus system (9-155 g/l) and BN II (9.3 - 262 g/l) we found positive correlations (r = 0.928, p < 0.0001), and (r = 0.803; p < 0.001) but with median difference 144% and 156% (ICC 0.213). Within the comparison of MIg levels defined by capillary electrophoresis and IgMκ detected by SPAplus system (9 - 155 g/l) and BN II (9.3 - 262 g/l) we found positive correlations (r = 0.959, p < 0.0001) and (r = 0.830; p < 0.001) with median difference 144% and 133% (ICC 0.225). The difference rate was increasing with MIg concentration. Conclusions: The results of our analysis confirm that the assessment of high IgM concentrations using HLC pairs is accompanied with significant overestimation of the values. Despite relatively strong correlations between conventional electrophoretic methods and HLC assessment, the results exceed not only MIg levels but even total protein levels. The assessment of HLC at this time cannot be reliably used for the diagnostics and monitoring of patients with active WM and high levels of MIg.