A technique for the collection of capillary blood samples and their subsequent analysis is described. The procedure is especially intended for use in children, stomatologic patients and individuals under load test who are to be screened for the level of immunoglobulins. The applicability of this method was verified in a group of 71 patients treated for various chronic liver lesions at the Research Institute of Balneology. The aim was to avoid the distress of repeated venous blood samplings to patients who had to be examined for the dynamic of changes in their immunologic parameters. Totally, nine immunologic parameters were simultaneously determined in the capillary and venous blood of these patients. As confirmed by statistical processing of the data (t-test, F-test, correlation coefficients and regression line equations were calculated) both methods proved equally suitable in terms of the results yielded so that either of them can be safely used for routine human serum analyses. However, the use of the capillary blood technique turned out to cause less discomfort to the patients, and appears to be distinctly of advantage in children and stomatologic patients as well as in repeated blood samplings during load tests and examinations in animal experiments.

• MeSH
• dítě MeSH
• dospělí MeSH
• imunoglobulin A analýza   MeSH
• imunoglobulin G analýza   MeSH
• imunoglobulin M analýza   MeSH
• imunoglobuliny analýza   MeSH
• kapiláry MeSH
• krevní proteiny analýza   MeSH
• lidé MeSH
• odběr vzorku krve * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH
• Názvy látek
• imunoglobulin A MeSH
• imunoglobulin G MeSH
• imunoglobulin M MeSH
• imunoglobuliny MeSH
• krevní proteiny MeSH

INTRODUCTION: The differentiation between extra- and intracellular production of reactive oxygen species (ROS) in whole blood was measured by luminol- and isoluminol-enhanced chemiluminescence (CL). METHODS: Azide (total CL inhibition), azide + horseradish peroxidase (HRP, restoring extracellular CL), superoxide dismutase + catalase (depleting extracellular ROS) and HRP (enhancing extracellular CL) were used to modulate luminol- and isoluminol-enhanced CL (10(-6) -10(-3) m luminophores) of 125× diluted whole blood which was activated by both calcium ionophore A23187 (Ca-I) and opsonized zymosan particles (OZP) separately. RESULTS: Both activators stimulated intra- and extracellular production of ROS. Luminol-enhanced CL of Ca-I-activated samples detected the intracellular ROS, and with the addition of HRP detected the extracellular CL as well. CL enhanced with isoluminol in concentrations of 10(-4) m or less was mostly extracellular. There was a mixture of intra- and extracellular CL in OZP-activated samples, probably because of the ingestion of luminophore molecules. CONCLUSION: Measurement of Ca-I-activated CL enhanced with 10(-4) m luminol is recommended for the detection of intracellular ROS. The addition of HRP leads to the detection of overall ROS production while the OZP-activated system with its addition of HRP can only be used to detect overall ROS production. Ca-I-activated CL enhanced with 10(-4) m isoluminol and with addition of HRP is recommended for the detection of extracellular CL.

• MeSH
• krev MeSH
• krevní buňky MeSH
• lidé MeSH
• luminiscence * MeSH
• luminiscenční měření metody   MeSH
• luminol analogy a deriváty   MeSH
• reaktivní formy kyslíku analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• isoluminol MeSH Prohlížeč
• luminol MeSH
• reaktivní formy kyslíku MeSH

A novel concept for highly versatile automated analyses of dried blood spot (DBS) samples by commercial capillary electrophoresis (CE) is presented. Two interchangeable CE cartridges with different fused-silica capillaries were used for the DBS elutions and the DBS eluate analyses, respectively. The application of one CE cartridge with a wide-bore capillary reduced DBS processing times to a minimum (1-2 min per sample) while fitting the other CE cartridge with a narrow-bore capillary served for highly efficient CE analyses. A comprehensive investigation of major variables affecting liquid handling in CE (capillary length, internal diameter, and temperature) was carried out with the aim of optimizing both procedures and enabling their maximum flexibility. The application of two CE cartridges also enabled the employment of CE detectors with different instrumental set-ups and/or principles as was demonstrated by the optical detection of nonsteroidal anti-inflammatory drugs (NSAIDs) and the conductivity detection of amino acids (AAs). The presented methods were optimized for the automated CE analyses of 36 DBS samples formed by a volumetric collection of 5 μL of capillary blood onto Whatman 903 discs and processed by direct in-vial elution using the CE instrument. The precision of liquid transfers for the automated DBS elutions was better than 0.9% and the precision of CE analyses did not exceed 5.1 and 12.3% for the determination of NSAIDs and AAs, respectively. Both methods were linear (R2 ≥ 0.996) over the therapeutic (NSAIDs) and the endogenous (AAs) concentration ranges, had limits of quantification below the typical analyte concentrations in human blood, and achieved sample throughputs of more than 6 DBSs per hour.

• MeSH
• aminokyseliny * MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární * metody   MeSH
• kapiláry MeSH
• lidé MeSH
• test suché kapky krve metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• aminokyseliny * MeSH

The unique feature of nuclear accidents with neutron exposure is the induced radioactivity in body tissues. For dosimetry purposes, the most important stable isotopes occurring in human body, which can be activated by neutrons, are 23 Na and 32 S. The respective activation reactions are as follows:23Na(n,γ)24Na and32S(n,p)32P. While sodium occurs in human blood, sulfur is present in human hair. In order to verify the practical feasibility of this dosimetry technique in conditions of our laboratory, samples of human blood and hair were irradiated in a channel of a training reactor VR-1.24Na activity was measured by gamma-ray spectrometry.32P activity in hair was measured by means of a proportional counter. Based on neutron-spectrum calculation, relationships between neutron dose and induced activity were derived for both blood and hair.

• MeSH
• dávka záření MeSH
• krev účinky záření   MeSH
• lidé MeSH
• neutrony * MeSH
• radioaktivita MeSH
• radioizotopy fosforu analýza   MeSH
• radioizotopy sodíku MeSH
• radiometrie metody   MeSH
• síra analýza   MeSH
• sodík analýza   MeSH
• únik radioaktivních látek MeSH
• vlasy, chlupy účinky záření   MeSH
• záření gama MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• Názvy látek
• Phosphorus-32 MeSH Prohlížeč
• radioizotopy fosforu MeSH
• radioizotopy sodíku MeSH
• síra MeSH
• sodík MeSH

BACKGROUND: The correct methodology of thrombelastography might be influenced by elapsing time. In our study we investigated kaolin activated citrated samples together with non-activated citrated samples in relation to the elapsed times of 0, 15 and 30 minutes to compare both methods and to find out if there is an impact of time on results of thrombelastography. METHODS: Blood samples obtained from 10 healthy volunteers were analyzed after 0, 15 and 30 minutes from sampling with kaolin activation and without activation. Then the results were analysed and compared between the non-activated and the kaolin-activated method. RESULTS: All blood samples became more hypercoagulable with the time elapsing, both in non-activated and kaolin-activated samples and differences between both groups were found statistically and clinically significant after only 0 minutes. CONCLUSIONS: The non-activated citrated method seems to be reliable and suitable for thrombelastography in non-emergency cases (planned surgical procedures) when we have time to wait 15-30 minutes to get results. In urgent situations a rapid thrombelastography test should be preferred. Although the kaolin-activated method can also be used, results must be interpreted with caution.

• MeSH
• analýza rozptylu MeSH
• časové faktory MeSH
• citráty farmakologie   MeSH
• kaolin farmakologie   MeSH
• koagulancia farmakologie   MeSH
• lidé MeSH
• odběr vzorku krve metody   MeSH
• tromboelastografie metody   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• klinické zkoušky MeSH
• srovnávací studie MeSH
• Názvy látek
• citráty MeSH
• kaolin MeSH
• koagulancia MeSH

A method is described for the determination of total glutathione (TGSH) in dried blood spot (DBS) samples using high-performance liquid chromatography with fluorescence detection. Whole blood and DBS samples were obtained from a group of blood donors. After GSH reduction with dithiothreitol and protein precipitation with ethanol, the samples were derivatized with naphthalene-2,3-dicarboxaldehyde to form a very stable, highly fluorescent derivative. For the separation, a reversed phase HPLC method was used. The mixture of ethanol and deionized water (8 : 92, v/v) was used as a mobile phase. The analytical performance of this method was satisfactory: the intra- and interassay coefficients of variation were below 10%. Quantitative recoveries from spiked DBS samples were between 98.3 and 103.6%. The presented method is inexpensive and suitable for clinical testing purposes.

• MeSH
• dospělí MeSH
• glutathion krev   chemie   MeSH
• lidé středního věku MeSH
• lidé MeSH
• limita detekce MeSH
• lineární modely MeSH
• methanol MeSH
• mladiství MeSH
• mladý dospělý MeSH
• reprodukovatelnost výsledků MeSH
• test suché kapky krve metody   MeSH
• vysokoúčinná kapalinová chromatografie metody   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• glutathion MeSH
• methanol MeSH

Polymer inclusion membranes (PIMs) have several important features, i.e., PIMs are dry and non-porous membranes, which can be prepared ahead of use and stored without noticeable deterioration in extraction performance. In this contribution, in-line coupling of microextractions across PIMs to a separation method for clinical purposes was demonstrated for the first time. Formate (the major metabolite in methanol poisoning) was determined in undiluted human serum and whole blood by capillary zone electrophoresis (CZE) with simultaneous capacitively coupled contactless conductivity detection (C(4)D) and UV-Vis detection. A purpose-made microextraction device with PIM was coupled to a commercial CZE instrument in order to ensure complete automation of the entire analytical procedure, i.e., of formate extraction, injection, CZE separation and quantification. PIMs for formate extractions consisted of 60% (w/w) cellulose triacetate as base polymer and 40% (w/w) Aliquat™ 336 as anion carrier. The method was characterized by good repeatability of peak areas (≤7.0%) and migration times (≤0.8%) and by good linearity of calibration curves (r(2) = 0.993-0.999). Limits of detection in various matrices ranged from 15 to 54 μM for C(4)D and from 200 to 635 μM for UV-Vis detection and were sufficiently low to clearly distinguish between endogenous and toxic levels of formate in healthy and methanol intoxicated individuals. In addition, PIMs proved that they may act as phase interfaces with excellent long-term stability since once prepared, they retained their extractions properties for, at least, two months of storage.

• Klíčová slova
• Blood samples, Capillary electrophoresis, Methanol intoxication, Microextraction, Polymer inclusion membranes,
• MeSH
• celulosa analogy a deriváty   chemie   MeSH
• design vybavení MeSH
• elektroforéza kapilární ekonomika   přístrojové vybavení   metody   MeSH
• formiáty krev   izolace a purifikace   MeSH
• lidé MeSH
• limita detekce MeSH
• membrány umělé * MeSH
• methanol otrava   MeSH
• mikroextrakce na pevné fázi ekonomika   přístrojové vybavení   metody   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Názvy látek
• cellulose triacetate MeSH Prohlížeč
• celulosa MeSH
• formiáty MeSH
• formic acid MeSH Prohlížeč
• membrány umělé * MeSH
• methanol MeSH

There is an increasing need for proper quality control tools in the pre-analytical phase of the molecular diagnostic workflow. The aim of the present study was to identify biomarkers for monitoring pre-analytical mRNA quality variations in two different types of blood collection tubes, K2EDTA (EDTA) tubes and PAXgene Blood RNA Tubes (PAXgene tubes). These tubes are extensively used both in the diagnostic setting as well as for research biobank samples. Blood specimens collected in the two different blood collection tubes were stored for varying times at different temperatures, and microarray analysis was performed on resultant extracted RNA. A large set of potential mRNA quality biomarkers for monitoring post-phlebotomy gene expression changes and mRNA degradation in blood was identified. qPCR assays for the potential biomarkers and a set of relevant reference genes were generated and used to pre-validate a sub-set of the selected biomarkers. The assay precision of the potential qPCR based biomarkers was determined, and a final validation of the selected quality biomarkers using the developed qPCR assays and blood samples from 60 healthy additional subjects was performed. In total, four mRNA quality biomarkers (USP32, LMNA, FOSB, TNRFSF10C) were successfully validated. We suggest here the use of these blood mRNA quality biomarkers for validating an experimental pre-analytical workflow. These biomarkers were further evaluated in the 2nd ring trial of the SPIDIA-RNA Program which demonstrated that these biomarkers can be used as quality control tools for mRNA analyses from blood samples.

• MeSH
• lidé MeSH
• messenger RNA krev   chemie   genetika   izolace a purifikace   MeSH
• odběr vzorku krve MeSH
• polymerázová řetězová reakce MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• stabilita RNA MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• validační studie MeSH
• Názvy látek
• messenger RNA MeSH

In this biomonitoring study, we evaluated the concentrations of 8 polychlorinated biphenyls (PCBs), 11 organochlorinated pesticides (OCPs), 33 brominated flame retardants (BFRs), 7 novel brominated and chlorinated flame retardants (novel FRs) and 30 per- and polyfluoroalkylated substances (PFAS) in human serum samples (n = 274). A total of 89 persistent organic pollutants (POPs) were measured in blood serum samples of city policemen living in three large cities and their adjacent areas (Ostrava, Prague, and Ceske Budejovice) in the Czech Republic. All samples were collected during the year 2019 in two sampling periods (spring and autumn). The identification/quantification of PCBs, OCPs, BFRs, novel FRs and PFAS was performed by means of gas chromatography coupled to (tandem) mass spectrometry (GC-MS/(MS)) and ultra-high performance liquid chromatography coupled to triple quadrupole tandem mass spectrometry (UHPLC-MS/MS). The most frequently detected pollutants were perfluorooctanoic acid (PFOA), perfluorononanoic acid (PFNA), perfluorodecanoic acid (PFDA), perfluorooctanesulfonate (PFOS), perfluorohexanesulfonate (PFHxS), 2,2',3,4,4',5'-hexachlorobiphenyl (CB 138), 2,2',4,4',5,5'-hexachlorobiphenyl (CB 153), 2,2',3,3',4,4',5-heptachlorobiphenyl (CB 170), 2,2',3,4,4',5,5'-heptachlorobiphenyl (CB 180), hexachlorobenzene (HCB), and p,p'-dichlorodiphenyldichloroethylene (p,p'-DDE) quantified in 100% of serum samples. In the serum samples, the concentrations of determined POPs were in the range of 0.108-900 ng g-1 lipid weight (lw) for PCBs, 0.106-1016 ng g-1 lw for OCPs, <0.1-618 ng g-1 lw for FRs and <0.01-18.3 ng mL-1 for PFAS, respectively. Locality, sampling season, and age were significantly associated with several POP concentrations. One of the important conclusions was that within the spring sampling period, statistically significant higher concentrations of CB 170 and CB 180 were observed in the samples from Ostrava (industrial area) compared to Prague and Ceske Budejovice. Older policemen had higher concentrations of five PCBs and two OCPs in blood serum.

• Klíčová slova
• City policemen, Czech Republic, Human biomonitoring, POPs, Serum,
• MeSH
• biologický monitoring MeSH
• chlorované uhlovodíky * analýza   MeSH
• látky znečišťující životní prostředí * analýza   MeSH
• lidé MeSH
• monitorování životního prostředí MeSH
• pesticidy * analýza   MeSH
• plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
• polychlorované bifenyly * analýza   MeSH
• sérum chemie   MeSH
• tandemová hmotnostní spektrometrie MeSH
• velkoměsta MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Česká republika MeSH
• velkoměsta MeSH
• Názvy látek
• chlorované uhlovodíky * MeSH
• látky znečišťující životní prostředí * MeSH
• pesticidy * MeSH
• polychlorované bifenyly * MeSH

The pathogenesis of prostate cancer (CaP) involves alterations in a gene structure of the androgen receptor (AR). The single nucleotide polymorphism AR-E211 G > A localized in exon 1 of the AR gene (G1733A) was detected using direct polymerase chain reaction and restriction digestion (PCR-RFLP) method on blood and tissue samples without prior DNA isolation. We used blood samples of patients with a diagnosis of benign prostatic hyperplasia (BPH) or CaP. From monitored group of CaP patients were selected specimen in formalin-fixed paraffin-embedded tissue blocks with morphology of BPH and CaP. The main objective of our study was to develop a method based the direct PCR-RFLP analysis from blood and tissue without prior DNA isolation for faster genotyping analysis of a large number of samples. We found no statistically significant differences in allelic % of the AR-E211 G > A polymorphism between BPH and CaP patients (p ≤ 0.8462). Genotyping of the AR-E211 G > A variant in blood was not identical with tumor tissue genotyping analysis. Significant agreement between blood and tissue AR-E211 G > A polymorphism only in non-tumor tissue focus was confirmed. Although we analyzed a limited number of the tissue samples, we suppose that a presence of the minor allele A may be associated with cancer transformation-induced changes of the modified AR gene.

• MeSH
• alely MeSH
• androgenní receptory krev   genetika   metabolismus   MeSH
• biochemická analýza krve metody   MeSH
• DNA analýza   genetika   MeSH
• dospělí MeSH
• genotyp MeSH
• hyperplazie prostaty krev   dietoterapie   genetika   metabolismus   MeSH
• jednonukleotidový polymorfismus genetika   MeSH
• krev MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory prostaty krev   diagnóza   genetika   metabolismus   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• androgenní receptory MeSH
• AR protein, human MeSH Prohlížeč
• DNA MeSH