A fast method for the determination of tropane alkaloids, using a portable CE instrument with a capacitively coupled contactless conductivity detector (CE-C4D) was developed and validated for determination of atropine and scopolamine in seeds from Solanaceae family plants. Separation was obtained within 5 min, using an optimized background electrolyte consisting of 0.5 M acetic acid with 0.25% (w/v) β-CD. The limit of detection and quantification was 0.5 µg/mL and 1.5 µg/mL, respectively, for both atropine and scopolamine. The developed method was validated with the following parameters-precision (CV): 1.07-2.08%, accuracy of the assay (recovery, RE): 101.0-102.7% and matrix effect (ME): 92.99-94.23%. Moreover, the optimized CE-C4D method was applied to the analysis of plant extracts and pharmaceuticals, proving its applicability and accuracy.

• Klíčová slova
• CE-C4D, atropine, capillary electrophoresis, conductivity detection, scopolamine, tropane alkaloids,
• MeSH
• alkaloidy Solanaceí analýza   MeSH
• atropin analýza   MeSH
• elektroforéza kapilární MeSH
• limita detekce MeSH
• skopolamin analýza   MeSH
• Solanaceae chemie   MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• alkaloidy Solanaceí MeSH
• atropin MeSH
• skopolamin MeSH

The role of antibodies in rejection of transplanted kidneys was the subject of debate at the last two Banff meetings and in medical journals. Diffuse C4d positive staining of peritubular capillaries (PTCs) was recognized as a marker of antibody-mediated rejection and this morphological feature was included in the updated Banff schema. At the same time polyomavirus infection of the renal allografts has been reported more frequently and is emerging as an important cause of renal allograft dysfunction and graft loss. At the present time, BK-virus nephropathy (BKN) represents the most common viral disease affecting renal allografts. BKN was identified in 6 patients in 12 biopsies and 2 graft nephrectomy specimens of 1115 biopsies between September 2000 and December 2003. Definite virus identification was done by immunohistochemistry. The reason for graft nephrectomies was graft failure due to BKN in a recipient after kidney-pancreas transplantation with good function of his pancreas graft and the necessity of continuing immunosuppression. Detection of C4d deposits was performed by immunofluorescence or by immunohistochemistry. In graftectomy samples C4d detection was performed by immunohistochemistry and retrospectively in all cases of BKN. Focal C4d positive PTCs and BKN were found simultaneously in 9 of 12 needle biopsies and in both graft nephrectomy samples. Detection of C4d by immunohistochemistry disclosed focal C4d positive staining in kidney tissue but diffuse in the sites where BK-virus inclusions in tubular epithelial cells were found. The complement system is part of the host defense response and is crucial to our natural ability to ward off infection. In cases of BKN, virus likely gains access to the bloodstream through injured tubular walls and via PTCs. Vascular endothelium in the PTCs represents a potential target antigen for alloresponse, and simultaneously possibly represents an imprint of complement activation or complement production in the places with BK-virus infection.

• MeSH
• infekce onkogenními viry imunologie   MeSH
• komplement C4b analýza   MeSH
• ledviny imunologie   MeSH
• lidé MeSH
• nemoci ledvin imunologie   MeSH
• peptidové fragmenty analýza   MeSH
• polyomavirové infekce imunologie   MeSH
• rejekce štěpu etiologie   MeSH
• transplantace ledvin * imunologie   MeSH
• virus BK * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• complement C4d MeSH Prohlížeč
• komplement C4b MeSH
• peptidové fragmenty MeSH

A simple and fast method for the analysis of lactate from a single drop of blood was developed. The finger-prick whole blood sample (10 µL) was diluted (1:20) with a 7% (w/v) solution of [tris(hydroxymethyl)methylamino] propanesulfonic acid and applied to a blood plasma separation device. The device accommodates a membrane sandwich composed of an asymmetric polysulfone membrane and a supporting textile membrane that allows the collection of blood plasma into a narrow glass capillary in less than 20 s. Separated and simultaneously diluted blood plasma was directly injected into a capillary electrophoresis instrument with a contactless conductivity detector (CE-C4D) and analyzed in less than one minute. A separation electrolyte consisted of 10 mmol/L l-histidine, 15 mmol/L dl-glutamic acid, and 30 µmol/L cetyltrimethylammonium bromide. The whole procedure starting from the finger-prick sampling until the CE-C4D analysis was finished, took less than 5 min and was suitable for monitoring lactate increase in blood plasma during incremental cycling exercise. The observed lactate increase during the experiments measured by the developed CE-C4D method correlated well with the results from a hand-held lactate analyzer (R = 0.9882). The advantage of the developed CE method is the speed, significant savings per analysis, and the possibility to analyze other compounds from blood plasma.

• Klíčová slova
• CE-C4D, blood plasma separation, capillary electrophoresis, lactate, training assessment,
• MeSH
• cvičení * MeSH
• elektrická vodivost * MeSH
• elektroforéza kapilární metody   MeSH
• kyselina mléčná krev   MeSH
• lidé MeSH
• limita detekce MeSH
• sportovci psychologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• kyselina mléčná MeSH

UNLABELLED: The aim of our retrospective study was to evaluate the clinical significance of measurement of the soluble CD30 (sCD30) molecule for the prediction of antibody-mediated (humoral) rejection (HR). Sixty-two kidney transplant recipients (thirty-one C4d-positive and thirty-one C4d-negative patients) were included into the study. Soluble CD30 levels were evaluated before transplantation and during periods of graft function deterioration. The median concentrations of the sCD30 molecule were identical in C4d-positive and C4d-negative patients before and after transplantation (65.5 vs. 65.0 and 28.2 vs. 36.0 U/ml, respectively). C4d+ patients who developed DSA de novo had a tendency to have higher sCD30 levels before transplantation (80.7+/-53.6 U/ml, n=8) compared with C4d-negative patients (65.0+/-33.4 U/ml, n=15). Soluble CD30 levels were evaluated as positive and negative (>or=100 U/ml and <100 U/ml respectively) and the sensitivity, specificity and accuracy of sCD30 estimation with regard to finding C4d deposits in peritubular capillaries were determined. The sensitivity of sCD30+ testing was generally below 40%, while the specificity of the test, i.e. the likelihood that if sCD30 testing is negative, C4d deposits would be absent, was 82%. C4d+ patients who developed DSA de novo were evaluated separately; the specificity of sCD30 testing for the incidence of HR in this cohort was 86%. CONCLUSION: We could not confirm in our study that high sCD30 levels (>or=100 U/ml) might be predictive for the incidence of HR. Negative sCD30 values might be however helpful for identifying patients with a low risk for development of DSA and antibody-mediated rejection.

• MeSH
• antigen Ki-1 analýza   MeSH
• dospělí MeSH
• homologní transplantace MeSH
• komplement C4b analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• peptidové fragmenty analýza   MeSH
• protilátky imunologie   MeSH
• rejekce štěpu imunologie   MeSH
• senioři MeSH
• transplantace ledvin imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• antigen Ki-1 MeSH
• complement C4d MeSH Prohlížeč
• komplement C4b MeSH
• peptidové fragmenty MeSH
• protilátky MeSH

Donor-specific antibodies (DSA) cause antibody-mediated rejection (AMR); however, their pathogenic role has not yet been adequately investigated after liver transplantation. The aim of our study was to analyse the clinical significance of DSA and complement-binding DSA for the prediction of AMR after liver transplantation. Our cohort included 120 liver recipients with assessed protocol biopsies one year post-transplant. All patients had defined HLA-specific and complement-binding (C1q + and C3d+) antibodies before and in regular intervals after transplantation. The incidence of DSA was evaluated in relation with clinical and histopathological data in the liver allografts. A higher occurrence of acute AMR was observed in recipients with preformed complement-binding DSA to HLA Class I antigens. Patients who developed chronic AMR had more frequently de novo-produced antibodies against HLA Class II antigens (P = 0.0002). A correlation was also found between de novo-formed C1q + and C3d+-binding antibodies to HLA Class II antigens and the development of chronic AMR (P = 0.043). Our study implies that preformed complement-binding DSA to HLA Class I antigens are related to increased risk of acute antibody-mediated rejection, while chronic AMR is more frequent in patients with de novo-produced antibodies to HLA Class II antigens after liver transplantation.

• Klíčová slova
• C4d deposits, HLA, complement-binding, donor-specific antibodies, rejection,
• MeSH
• dárci tkání MeSH
• HLA antigeny MeSH
• isoprotilátky MeSH
• komplement C1q MeSH
• lidé MeSH
• přežívání štěpu MeSH
• rejekce štěpu MeSH
• retrospektivní studie MeSH
• transplantace jater * škodlivé účinky   MeSH
• transplantace ledvin * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• HLA antigeny MeSH
• isoprotilátky MeSH
• komplement C1q MeSH

The aim of our study was to evaluate the relevance of complement-binding donor-specific antibodies (DSA) for prediction of antibody-mediated rejection (AMR) after liver transplantation. Sera from 123 liver transplant recipients were retrospectively defined for HLA specificity and complement-fixing activity using the single antigen beads, C1q and C3d techniques. Liver-recipients' sera were tested before transplantation, 3, 6 months and 1 year after transplantation. Patients were followed up for graft survival and rejection incidence for 1 year after transplantation. All patients with pretransplant complement-binding DSA developed severe AMR after transplantation, while three recipients out of four, who produced de novo complement-fixing DSA, developed AMR. Definition of DSA with respect to complement-fixing activity may provide clinically relevant information about the risk of AMR after liver transplantation.

• Klíčová slova
• C4d, HLA, antibodies, complement, crossmatch, liver transplantation, rejection,
• MeSH
• dárci tkání MeSH
• dítě MeSH
• dospělí MeSH
• homologní transplantace MeSH
• isoprotilátky krev   MeSH
• komplement C1q metabolismus   MeSH
• komplement C3d metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• následné studie MeSH
• přežívání štěpu * MeSH
• prognóza MeSH
• rejekce štěpu krev   diagnóza   imunologie   patologie   MeSH
• retrospektivní studie MeSH
• senioři MeSH
• testování histokompatibility metody   MeSH
• transplantace jater * MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mužské pohlaví MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• isoprotilátky MeSH
• komplement C1q MeSH
• komplement C3d MeSH

Contactless conductivity detection (C4D) as a universal detection technique plays an important role in combination with efficient electrophoretic separation carried out in capillaries (CE) or on microchips (ME) in the analysis of clinical samples. C4D is particularly sensitive in the quantification of low molecular weight biogenic substances such as inorganic cations and anions, amino acids, amines, low molecular weight organic acids, saccharides and many drugs such as antibiotics, analgesics, anaesthetics or antiepileptics. Biogenic substances are determined in CE/C4D or ME/C4D directly in their native form without derivatization and sample matrix treatment is often based only on dilution or addition of an organic solvent. The limit of detection for most CE/C4D determinations is at the micromolar concentration level, which is sufficient to monitor physiological or therapeutic levels of most of low molecular weight biogenic substances. Therefore, CE/C4D and ME/C4D are widely used for sequential monitoring of nutrients, metabolites and waste products at the level of individual tissues and organs, low-invasive detection of inborn errors of metabolism and cystic fibrosis, pharmacokinetic monitoring and therapeutic drug monitoring. Innovative trends such as electrophoretic stacking, microdialysis, electromembrane extraction, portable and disposable CE instruments and minimally invasive clinical sampling techniques are mentioned. A critical evaluation of the positives and negatives of this technique is presented, covering the main applications published over the last 10 years.

• Klíčová slova
• Capillary electrophoresis, Clinical analysis, Contactless conductivity detection, Electromembrane extraction, Microchip electrophoresis, Microdialysis,
• MeSH
• aminy MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární metody   MeSH
• elektroforéza mikročipová * metody   MeSH
• kapiláry MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• aminy MeSH

CE with capacitively coupled contactless detection (C4D) was used to determine 3-methylhistidine (3-MH) and 1-methylhistidine (1-MH). The C4D response to 3-MH was studied in a BGE consisting of 500 mM acetic acid and ammonia at varying concentration and the results were compared with the theory. Complete separation of a model mixture of 3-MH, 1-MH, and histidine (His) was attained in two optimized BGEs, one containing 500 mM HAc, 20 mM NH4OH, and 0.1 % m/v hydroxyethylcellulose (HEC), pH 3.4 (I) and the other consisting of 100 mM morpholinoethanesulfonic acid (MES), 25 mM LiOH, and 0.1 % m/v HEC, pH 5.5 (II). These optimized BGEs were tested in CE/C4D analyses of urine. Promising results were obtained for separation and determination of 3-MH, 1-MH, and His on a silicon microchip, using aluminum strips as the C4D electrodes; the three analytes were baseline-separated within less than 30 s with a separation channel effective length of 38 mm. The LOD were satisfactory and amounted to 26.4 microM for 3-MH and 18.3 microM for 1-MH.

• MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární metody   MeSH
• elektroforéza mikročipová přístrojové vybavení   metody   MeSH
• lidé MeSH
• methylhistidiny analýza   moč   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Názvy látek
• 1-methylhistidine MeSH Prohlížeč
• 3-methylhistidine MeSH Prohlížeč
• methylhistidiny MeSH

The view on the role of donor-specific antibodies in organ transplantation has been changed during the last several decades. Today, it is considered that the majority of cases of the late renal allograft dysfunction and loss are caused by the presence of donor-specific antibodies to HLA antigens. The real breakthrough in the diagnosis of antibody-mediated rejection was represented by the discovery of C4d, which enabled the determination of the diagnostic criteria of acute and later chronic antibody-mediated rejection. Although detection of C4d has been the cornerstone in the diagnosis of antibody-mediated rejection for over 10 years, it has become clear that some cases with similar morphological and clinical features do not have detectable C4d. Outcomes of key studies concerning presence of donor specific antibodies and morphological features in the graft biopsy samples resulted in the modification of Banff classification of 2013, which includes integrating C4d negative antibody-mediated rejection and also that acute vascular rejection (v1, v2) can be a part of the antibody-mediated rejection.

• Klíčová slova
• kidney allograft - antibody-mediated rejection - T-cell mediated rejection.,
• MeSH
• dárci tkání MeSH
• HLA antigeny MeSH
• komplement C4b imunologie   MeSH
• ledviny imunologie   patologie   MeSH
• lidé MeSH
• rejekce štěpu diagnóza   imunologie   MeSH
• transplantace ledvin škodlivé účinky   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• HLA antigeny MeSH
• komplement C4b MeSH

The paper provides a comprehensive survey of the use of capillary and microchip electrophoresis in combination with contactless conductivity detection (C4D) for the analysis of drinking water, beverages and foodstuffs. The introduction sets forth the fundamentals of conductivity detection anddescribes an axialC4Dversion. There is also a detailed discussion of the determination of inorganic ions, organic acids, fatty acids, amino acids, amines, carbohydrates, foreign substances and poisons from the standpoint of separation conditions, sample treatment and detection limits. Special attention is paid to the analysis of foodstuffs at microchips with emphasis on the employed material and connection of the microchip with the C4D. The review attempts to draw attention to modern trends, such as dual-opposite injection, field-enhanced sample injection, electromembrane extraction and on-line combination of microdialysis with CE. CE/C4D is characterised by high universality, high speed of analysis, simple sample preparation, small consumption of sample and other chemicals.

• Klíčová slova
• Beverages, Capillary electrophoresis, Contactless conductivity detection, Extraction, Food, Microchip electrophoresis,
• MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární MeSH
• elektroforéza mikročipová * MeSH
• ionty MeSH
• nápoje analýza   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• ionty MeSH