Autologous serum eye drops (ASEDs) are used as a treatment for severe dry eye disease. The concentration and stability of various growth factors in ASEDs is determinative for their efficiency. We therefore assessed the concentrations of transforming growth factor beta 1 (TGF-β1), epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF-1) in ASEDs following storage at 4-8, -20, -80 and -156 °C. Twenty % and 100% sera from eight healthy volunteers were analysed by the sandwich enzyme immunoassay at different time intervals up to seven months. The mean levels of TGF-β1 and EGF in undiluted and 20% serum did not differ significantly from the baseline levels in fresh serum for any storage conditions after 7 days at 4-8 °C, as well as after 4- and 7-month preservation at sub-zero temperatures. In 20% serum, no IGF-1 concentration decrease was found following 7 days of preservation at 4-8 °C. However, a decrease to 78 % and 81 % (P < 0.01) of baseline values was found in 20% serum after 4-month storage at -20 °C and 7-month storage at -156 °C, respectively. A more pronounced decrease in IGF-1 was observed in undiluted serum. All assessed growth factors present in 20% frozen serum remained stable for up to 7 months. The highest stability was achieved at -80 °C. At -20 and -156 °C, some decrease in IGF-1 occurred. Our results indicate that 20% ASEDs can be stored frozen up to 7 months under proper conditions.

• MeSH
• epidermální růstový faktor * MeSH
• insulinu podobný růstový faktor I * metabolismus   MeSH
• lidé MeSH
• oční roztoky MeSH
• sérum metabolismus   MeSH
• teplota MeSH
• transformující růstový faktor beta1 MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The aim of this study was to determine the effect of autologous serum (AS) eye drops on the density of human leucocyte antigen (HLA)-DR-positive epithelial cells and Langerhans cells on the ocular surface of patients with bilateral severe dry eye disease (DED) due to graft-versus-host disease (GvHD) or Sjögren's syndrome (SS). The study was conducted on 24 patients (48 eyes). AS was applied 6-10 times daily for 3 months together with regular artificial tear therapy. HLA-DR-positive cells were detected by direct immunocytochemistry on upper bulbar conjunctiva imprints obtained before and after treatment. The application of AS drops led to a statistically significant increase in the mean density of aberrant HLA-DR-positive conjunctival epithelial cells (p < 0.05) and HLA-DR-positive Langerhans cells (p < 0.05) in the GvHD group. Aberrant HLA-DR-positive epithelial cells in the SS group were decreased non-significantly. All patients reported a significant decrease in the Ocular Surface Disease Index (p < 0.01), which indicates improvement of the patient's subjective feelings after therapy. There was an expected but non-significant decrease of aberrant HLA-DR-positive conjunctival epithelial cells in the SS group only. However, the increased density of HLA-DR-positive cells, indicating slight subclinical inflammation, does not outweigh the positive effect of AS in patients with DED from GvHD.

• MeSH
• dospělí MeSH
• epitel metabolismus   MeSH
• epitelové buňky účinky léků   metabolismus   MeSH
• HLA-DR antigeny metabolismus   MeSH
• imunohistochemie metody   MeSH
• konjunktiva účinky léků   metabolismus   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nemoc štěpu proti hostiteli farmakoterapie   metabolismus   MeSH
• oční roztoky terapeutické užití   MeSH
• počet buněk metody   MeSH
• senioři MeSH
• sérum metabolismus   MeSH
• Sjögrenův syndrom farmakoterapie   metabolismus   MeSH
• syndromy suchého oka farmakoterapie   metabolismus   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

The Eurotransplant (The Eurotransplant International Foundation) acceptable mismatch programme has been shown to be a successful tool to enhance transplantation of highly sensitized patients(HSPs). However, patients with rare HLA phenotypes in relation to the Eurotransplant donor population remain on the waiting list. EUROSTAM is an European Union funded project to explore the feasibility of a Europe-wide acceptable mismatch programme enabling transplantation of HSPs with rare HLA phenotypes within their own organ exchange organization. The present study, which forms part of the EUROSTAM project, assesses the differences in the practices of the laboratories in different countries with respect to their HLA antibody profiling and risk adverseness. In the serum exchange exercises of 18 samples, a high level of variability has been shown in both assays and interpretation of results. In the data exchange exercise when all participants were given the same Luminex raw data for analysis, a high degree of consensus was reached where the median fluorescent intensity values of beads were <500 and >2000 for standard single antigen bead assays, or <500 and >5000 for assignment of acceptable mismatches. The risk adverseness analysis has showed distinct patterns of attitudes towards the perceived risks based on HLA antibody assay results, most probably influenced by the local protocols of the clinical transplant programme of each laboratory. In order to ensure fairness and maintain consistencies of organ exchange among partner transplant centres, a centralized facility will be instrumental for a uniform definition of acceptable mismatches.

• MeSH
• fluorescence MeSH
• imunizace * MeSH
• konsensus MeSH
• lidé MeSH
• sérum metabolismus   MeSH
• testování histokompatibility * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The storage time and storage temperature might affect stability of oxidative stress biomarkers, therefore, they have to be analyzed after long-term storage of serum samples. The stability of three biomarkers reflecting oxidative stress: reactive oxygen metabolites (ROM) for hydroperoxides, total thiol levels (TTL) for the redox status and biological antioxidant potency (BAP) for the antioxidant status, was investigated at several time points during 60 months of storage at -20 and -80 °C. Biomarkers ROM and BAP showed a very good stability during storage for 60 months at both temperatures. In addition, the correlation of the data after 60 months of storage compared with the starting data was very good with correlation coefficients >0.9. The TTL assay showed good results in serum samples stored at -80 °C, but not in samples stored at -20 °C. Serum samples for analysis of the set of oxidative stress biomarkers ROM, BAP and TTL can be stored up to 60 months at -80 °C. ROM and BAP can also be stored at -20 °C during this period. The present results are very important for the biomarker-related epidemiological studies that make use of biobanks with samples stored for many years and for new project planning, including sample storage conditions.

• MeSH
• biologické markery krev   MeSH
• lidé MeSH
• oxidační stres * MeSH
• sérum metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Circulating cell-free DNA (cfDNA) may be involved in immune response regulation. We studied the variations in abundance of telomeric sequences in plasma and serum in young healthy volunteers and the ability of cfDNA contained in these samples to co-activate the TNF-α m RNA expression in monocytes. We performed qPCR to determine relative telomere length (T/S ratios) in plasma, serum and whole blood of 36 volunteers. Using paired samples of plasma and serum and DNase treatment, we analysed the contribution of cfDNA to the co-activation of TNF-α mRNA expression in THP1 monocytic cell line. We found significant differences between paired plasma and serum samples in relative T/S ratios (median 1.38 ± 1.1 vs. 0.86 ± 0.25, respectively) and in total amounts of cfDNA and in estimated total amounts of telomeres which were significantly higher in serum than in plasma. TNF-α mRNA expression in THP1 cells increased significantly after DNase treatment of all samples used for stimulation. The highest TNF-α mRNA expressions were observed after stimulation with DNase treated serum samples. Our results suggest that the different content of telomeric sequences in plasma and serum may contribute to the tuning of immune response. Further studies of this interesting phenomenon are needed.

• MeSH
• deoxyribonukleasy metabolismus   MeSH
• homeostáza telomer MeSH
• imunita MeSH
• imunomodulace MeSH
• krevní plazma imunologie   metabolismus   MeSH
• lidé MeSH
• mladý dospělý MeSH
• monocyty fyziologie   MeSH
• sérum imunologie   metabolismus   MeSH
• telomery genetika   MeSH
• THP-1 buňky MeSH
• TNF-alfa genetika   metabolismus   MeSH
• upregulace MeSH
• volné cirkulující nukleové kyseliny genetika   imunologie   metabolismus   MeSH
• zdraví dobrovolníci pro lékařské studie MeSH
• Check Tag
• lidé MeSH
• mladý dospělý MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

In this work, a sensitive capillary electrophoresis method with laser induced fluorescence detection for determination of malondialdehyde in various biological fluids was developed. Malondialdehyde reacts with thiobarbituric acid under optimized conditions of pH=2, reaction time of 60min and temperature of 90°C, yielding an adduct that can be separated in a 50mM sodium borate background electrolyte at pH 9. The separation of the formed adduct was accomplished in less than 6min with limit of detection of 1.1nM due to the use of 532nm laser module, exactly matching the maximum excitation wavelength of the formed adduct. The developed method offers unprecedented sensitivity and was for the first time used for analysis of malondialdehyde in exhaled breath condensate. The method proved to be also applicable to other samples of biological fluids, such as blood plasma and saliva.

• MeSH
• dechové testy metody   MeSH
• elektroforéza kapilární metody   MeSH
• fluorescence MeSH
• lasery * MeSH
• lidé MeSH
• malondialdehyd analýza   MeSH
• sérum metabolismus   MeSH
• sliny metabolismus   MeSH
• vydechnutí * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Akútny svalový kompartment syndróm vzniká náhlym zvýšením tlaku v anatomickom priestore, ktorý je ohraničený málo poddajným obalom. Každá svalová skupina, ohraničená fasciou, prípadne aj kosťou, vytvára svalový kompartment, v ktorom sa môže zvýšiť tlak. Zvyšujúci sa intrakompartmentový tlak spôsobuje klesanie perfúzneho tlaku, svalové bunky trpia ischémiou s poruchou tvorby a utilizácie energetických substrátov. Porucha metabolizmu buniek je spočiatku funkčná a reverzibilná, časom progreduje do ireverzibilných zmien a končí myonekrózou. Obnovením prekrvenia sa síce odstráni lokálna ischémia, ale následný reperfúzny syndróm so sprievodným oxidačným stresom je ďalším zdrojom progresie lokálneho patologického procesu. Obnovením cirkulácie sa ložisko stáva súčasťou vnútorného prostredia, produkty myonekrózy a tiež v mieste lézie aktivované imunokompetentné bunky prúdia krvou do celého organizmu. Lokálne aktivovaný endotel je mohutným zdrojom mediátorov zápalu. Prozápalový stav indukuje rozvoj protizápalových regulačných mechanizmov a od ich vzájomnej inter­akcie závisí výsledný efekt lokálny aj celkový.

Acute compartment syndrome results from a pressure increase within an anatomically defined space delineated by a non-elastic surrounding. Every muscle group, encased by fascia/bone, presents a compartment with a potential to develop increased pressure under certain conditions. Increase in pressure within the compartment causes a decrease in perfusion pressure, leading to ischemia and pathological utilisation of energy substrates on the cellular level. Initially, the malfunction of cellular metabolism is functional and reversible. Later progress of these changes leads into irreversible myonecrosis. Restitution of blood supply reverses the local ischemia, however, the following reperfusion syndrome associated with oxidative stress, leads to further pathological sequelae. Re-established blood circulation carries the end-products of myonecrosis and activated immunocompetent cells from the site of lesion into the whole body. Furthermore, locally activated endothelium becomes a significant source of systemic inflammatory mediators. Pro-inflammatory conditions induce the response of anti-inflammatory regulatory mechanisms and their mutual interaction determines both local and systemic outcome of the disease.

• MeSH
• biochemická analýza krve MeSH
• buněčná membrána metabolismus   MeSH
• crush syndrom * metabolismus   patofyziologie   MeSH
• hyperfosfatemie MeSH
• hyperkalemie MeSH
• hyperurikemie MeSH
• hypokalcemie MeSH
• intracelulární prostor MeSH
• ischemie MeSH
• kompartment syndrom * patologie   MeSH
• lidé MeSH
• myoglobin MeSH
• nemoci svalů krev   metabolismus   patofyziologie   MeSH
• renální insuficience MeSH
• rhabdomyolýza etiologie   krev   metabolismus   patofyziologie   MeSH
• sérum metabolismus   MeSH
• svaly * anatomie a histologie   fyziologie   metabolismus   patofyziologie   MeSH
• zánět MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

• MeSH
• financování organizované MeSH
• insulinu podobný růstový faktor I analýza   metabolismus   MeSH
• kardiovaskulární nemoci krev   metabolismus   mortalita   MeSH
• lidé středního věku MeSH
• lidé MeSH
• nádory krev   metabolismus   mortalita   MeSH
• následné studie MeSH
• populace MeSH
• příčina smrti MeSH
• registrace MeSH
• senioři nad 80 let MeSH
• sérum metabolismus   MeSH
• Check Tag
• lidé středního věku MeSH
• lidé MeSH
• senioři nad 80 let MeSH
• ženské pohlaví MeSH
• Publikační typ
• souhrny MeSH
• Geografické názvy
• Nizozemsko MeSH

• MeSH
• adiponektin analýza   krev   MeSH
• dyslipidemie farmakoterapie   komplikace   MeSH
• experimenty na zvířatech MeSH
• financování organizované MeSH
• hypolipidemika škodlivé účinky   terapeutické užití   MeSH
• inzulinová rezistence MeSH
• krysa rodu rattus MeSH
• lidé MeSH
• mastné kyseliny analýza   metabolismus   MeSH
• metabolismus lipidů účinky léků   MeSH
• metabolismus sacharidů účinky léků   MeSH
• niacin aplikace a dávkování   farmakologie   škodlivé účinky   MeSH
• oxidace-redukce účinky léků   MeSH
• sérum metabolismus   účinky léků   MeSH
• statistika jako téma MeSH
• svaly účinky léků   MeSH
• zvířata MeSH
• Check Tag
• krysa rodu rattus MeSH
• lidé MeSH
• zvířata MeSH

Micro-electrodialysis (μED) and CE were combined for rapid pretreatment and subsequent determination of inorganic cations in biological samples. Combination of μED with CE greatly improved the analytical performance of the latter as the adsorption of high molecular weight compounds present in real samples on the inner capillary wall was eliminated. Fifty microliter of 80-fold diluted human body fluids such as plasma, serum and whole blood was used in the donor compartment of the μED system requiring less than 1 μL of the original body fluid per analysis. Inorganic cations that migrated through a cellulose acetate dialysis membrane with molecular weight cut-off value of 500 Da were collected in the acceptor solution and were then analyzed using CE-C⁴D. Baseline separation of inorganic cations was achieved in a BGE solution consisting of 12.5 mM maleic acid, 15 mM L-arginine and 3 mM 18-crown-6 at pH 5.5. Repeatability of the CE-C⁴D method was better than 0.5% and 2.5% for migration times and peak areas, respectively; limits of detection of all inorganic cations in the presence of 2 mM excess of Na(+) were around 1 μM and calibration curves were linear with correlation coefficients better than 0.998. Repeatability of the sample pretreatment procedure was calculated for six independent electrodialysis runs of artificial and real samples and was better than 11.8%. Recovery values between 96.3 and 110% were achieved for optimized electrodialysis conditions of standard solutions and real samples; lifetime of the dialysis membranes for pretreatment of real samples was estimated to 100 runs.

• MeSH
• elektrická vodivost MeSH
• elektroforéza kapilární metody   MeSH
• elektrolyty chemie   MeSH
• kationty analýza   krev   MeSH
• krev metabolismus   MeSH
• krevní plazma metabolismus   MeSH
• lidé MeSH
• mikrodialýza přístrojové vybavení   MeSH
• reprodukovatelnost výsledků MeSH
• roztoky chemie   MeSH
• sérum metabolismus   MeSH
• tělesné tekutiny chemie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH