A novel, easy to use and portable capillary electrophoretic instrument for injection of small volumes of biological fluids equipped with contactless conductivity detection was constructed. The instrument is lightweight (<5 kg), all necessary parts including a tablet computer are accommodated in a plastic briefcase with dimensions 20 cm × 33 cm × 17 cm (w × l × h), allows hydrodynamic injection of small sample volumes and can continuously operate for at least 10 hours. The semi-automated hydrodynamic sample injection is accomplished via a specially designed PMMA interface that is able to repeatedly inject sample aliquots from a sample volume as low as 10 μL, with repeatability of peak areas below 5%. The developed interface and the instrument were optimized for the injection of biological fluids. Practical utility was demonstrated on the determination of formate in blood serum samples from acute methanol intoxication patients and on the analysis of ionic profile (nitrosative stress markers, including nitrite and nitrate) in the exhaled breath condensate from one single exhalation.
- MeSH
- Breath Tests MeSH
- Nitrates analysis MeSH
- Nitrites analysis MeSH
- Electrophoresis, Capillary instrumentation methods MeSH
- Feces chemistry MeSH
- Formates blood MeSH
- Humans MeSH
- Methanol poisoning MeSH
- Body Fluids chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Nitrites represent a serious health problem especially if present in drinking water for infants. On the other hand, they are used in the treatment of some cardiovascular diseases. The article deals with analytical methods used for the determination of nitrites in various samples.
- MeSH
- Chemistry Techniques, Analytical * MeSH
- Chromatography, Ion Exchange methods utilization MeSH
- Child MeSH
- Adult MeSH
- Nitrites * analysis chemistry metabolism adverse effects MeSH
- Electrochemical Techniques methods utilization MeSH
- Carcinogens, Environmental analysis MeSH
- Food Contamination analysis MeSH
- Humans MeSH
- Methemoglobinemia etiology MeSH
- Infant, Newborn MeSH
- Drinking Water analysis MeSH
- Spectrophotometry methods utilization MeSH
- Therapeutic Equivalency MeSH
- Titrimetry methods utilization MeSH
- Environmental Pollution analysis MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Humans MeSH
- Infant, Newborn MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
The analysis of an ionic profile of exhaled breath condensate (EBC) by capillary electrophoresis with contactless conductivity detection and double opposite end injection, is demonstrated. A miniature sampler made from a 2 ml syringe and an aluminium cooling cylinder was used for the fast collection of EBC (under one minute). Analysis of the collected EBC was performed in a 60 mM 2-(N-morpholino)ethanesulfonic acid, 60 mM L-histidine background electrolyte with 30 µM cetyltrimethylammonium bromide and 2 mM 18-crown-6 at pH 6, and excellent repeatability of migration times (RSD <1.3% (n = 7)) and peak areas (RSD < 7% (n = 7)) of 14 ions (inorganic anions, cations and organic acids) was obtained. It is demonstrated that the analysis of EBC samples obtained from patients with various respiratory diseases (chronic obstructive pulmonary disease, asthma, pulmonary fibrosis, sarcoidosis, cystic fibrosis) is possible in less than five minutes and the ionic profile can be compared with the group of healthy individuals. The analysis of the ionic profile of EBC samples provides a set of data in which statistically significant differences among the groups of patients could be observed for several clinically relevant anions (nitrite, nitrate, acetate, lactate). The developed collection system and method provides a highly reproducible and fast way of collecting and analyzing EBC, with future applicability in point-of-care diagnostics.
- MeSH
- Acetates analysis metabolism MeSH
- Ammonium Compounds analysis metabolism MeSH
- Asthma metabolism MeSH
- Butyrates analysis metabolism MeSH
- Chlorides analysis metabolism MeSH
- Pulmonary Disease, Chronic Obstructive metabolism MeSH
- Crown Ethers MeSH
- Cystic Fibrosis metabolism MeSH
- Breath Tests instrumentation methods MeSH
- Adult MeSH
- Potassium analysis metabolism MeSH
- Nitrates analysis metabolism MeSH
- Nitrites analysis metabolism MeSH
- Electrophoresis, Capillary MeSH
- Magnesium analysis metabolism MeSH
- Ions MeSH
- Hydrogen-Ion Concentration MeSH
- Lactic Acid analysis metabolism MeSH
- Middle Aged MeSH
- Humans MeSH
- Pulmonary Fibrosis metabolism MeSH
- Sarcoidosis, Pulmonary metabolism MeSH
- Propionates analysis metabolism MeSH
- Aged MeSH
- Sulfates analysis metabolism MeSH
- Sodium analysis metabolism MeSH
- Calcium analysis metabolism MeSH
- Check Tag
- Adult MeSH
- Middle Aged MeSH
- Humans MeSH
- Male MeSH
- Aged MeSH
- Female MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
Cílem naší studie bylo zjistit, zda léčba tiotropiem ovlivní nitrační stres u nemocných s CHOPN. Jedná se o otevřenou studii. Koncentrace dusitanů a dusičnanů byly měřeny v kondenzátu vydechovaného vzduchu (KVV). Odběry KVV byly prováděny před zahájením léčby, po 4 a po 12ti týdnech léčby. Koncentrace dusitanů se po 4 ani po 12ti týdenní léčbě v porovnání s koncentrací před léčbou signifikantně nezměnily. Koncentrace dusičnanů byla po 12ti týdnech léčby tiotropiem signifikantně menší než před léčbou. Podávání tiotropia po dobu 12ti týdnů nemocným s CHOPN dokáže zmírnit nitrační stres (p=0,010).
The study aimed at determining whether tiotropium therapy influences nitrosative stress in patients with COPD. This is an open study. Nitrite and nitrate concentrations were measured in exhaled breath concentrate (EBC). EBC was collected before therapy initiation and after 4 and 12 weeks of treatment. Nitrite concentrations did not significantly change after 4 or 12 weeks of treatment when compared with concentrations prior to the therapy. After 12 weeks of tiotropium therapy, nitrate concentrations were significantly lower than those before the therapy. Twelve-week administration of tiotropium to COPD patients is able to decrease nitrosative stress (p = 0.010),
- MeSH
- Pulmonary Disease, Chronic Obstructive drug therapy MeSH
- Breath Tests methods MeSH
- Nitrates analysis diagnostic use MeSH
- Nitrites analysis diagnostic use MeSH
- Middle Aged MeSH
- Humans MeSH
- Oxidative Stress drug effects MeSH
- Nitrogen Oxides metabolism adverse effects MeSH
- Aged MeSH
- Scopolamine Derivatives administration & dosage pharmacology MeSH
- Check Tag
- Middle Aged MeSH
- Humans MeSH
- Aged MeSH
Byla popsána nová skupina tzv. Toll-like receptors (TLR), celkem 12 typů. Tyto receptory jsou schopny identifikovat nukleové kyseliny, především různé formy bakteriálních lipopolysacharidů uvolněné z mikrobů po fagocytaci gramnegativních baktérií. V experimentu se ukázalo, že TLR neexprimuje v oblasti dutého systému ledvin. Tubulární buňky nejsou schopné exprese TLR4 a neprodukují žádné CC-chemokiny. TLR4 je zodpovědný za výskyt pyurie a tvorbu abscesů v ledvině. Aplikace těchto experimentů do klinické medicíny není zatím příliš jasná.
- MeSH
- Anti-Bacterial Agents therapeutic use MeSH
- Early Diagnosis MeSH
- Diagnostic Techniques, Urological classification utilization MeSH
- Child MeSH
- Adult MeSH
- Nitrites analysis diagnostic use urine MeSH
- Drug Therapy methods MeSH
- Urinary Tract Infections diagnosis complications therapy MeSH
- Interleukin-8 analysis diagnostic use urine MeSH
- Infant MeSH
- Creatinine administration & dosage diagnostic use MeSH
- Humans MeSH
- Adolescent MeSH
- Reagent Strips administration & dosage therapeutic use MeSH
- Toll-Like Receptor 4 analysis genetics MeSH
- Toll-Like Receptors analysis MeSH
- Check Tag
- Child MeSH
- Adult MeSH
- Infant MeSH
- Humans MeSH
- Adolescent MeSH
- Male MeSH
- Female MeSH
In this study, we realized the continual and long-term electrochemical detection of NO production by stimulated macrophages using modified porphyrinic microsensor. The NO release from RAW 264.7 cells stimulated by lipopolysaccharide started 5 h after the lipopolysaccharide administration. After reaching its maximum at the sixth hour, the stable level of NO production was observed between the seventh and 12th hour of the experiment. This phase was followed by a gradual decline in NO production. A close correlation between the NO signal detected with microelectrode and nitrite accumulation, which had been determined in supernatants removed from stimulated cells, was observed. This finding was utilized for the calibration of the electrochemical experiment. The presence of iNOS enzyme, which constitutes a main requirement for NO production by stimulated macrophages, was confirmed by Western blot analysis of iNOS protein expression at key time points of the corresponding electrochemical experiment. The capability of our microsensor to instantaneously monitor the changes in the NO production by stimulated RAW 264.7 cells was demonstrated by the immediate decrease in the signal due to NO as a response to the addition of iNOS inhibitor into the cell culture medium.
- MeSH
- Cell Line MeSH
- Time Factors MeSH
- Nitrites analysis chemistry metabolism MeSH
- Electrochemical Techniques methods instrumentation MeSH
- Lipopolysaccharides pharmacology MeSH
- Macrophages chemistry metabolism secretion drug effects MeSH
- Mice MeSH
- Nitric Oxide analysis biosynthesis chemistry secretion MeSH
- Nitric Oxide Synthase Type II metabolism MeSH
- Thiazines MeSH
- Animals MeSH
- Check Tag
- Mice MeSH
- Animals MeSH
- Publication type
- Research Support, Non-U.S. Gov't MeSH
Zajímalo nás, zda i u závažných exacerbací vyžadujících hospitalizaci dojde po jejich odeznění ke zmírnění oxidačního stresu. K hodnocení jsme vybrali tři látky podílející se na oxidačním stresu – nitrity (NO2-), nitráty (NO3-), což jsou látky vznikající přeměnou oxidu dusnatého (NO), a malondialdehyd, který je produktem peroxidace lipidů. Koncentrace NO2- i NO3- byly měřeny v kondenzátu vydechovaného vzduchu (KVV), který byl získán na přístroji EcoScreen (VIASYS HealthCare GmbH, Hoechberg, SRN). Soubor tvoří celkem 29 osob, z toho 18 mužů (62,1 %) a 11 (37,9 %) žen hospitalizovaných pro exacerbaci CHOPN. Medián koncentrace NO2- (umol/l) při přijetí byl 5,11; při propuštění 6,58; medián NO3- (umol/l) byl 42,3 resp. 53,0; medián koncentrace malondialdehydu (nmol/l) byl při přijetí 30,5; při propuštění 17,7. U žádného ze sledovaných parametrů nedošlo k signifikantnímu poklesu. Výsledky naší práce dokládají, že u nemocných hospitalizovaných pro exacerbaci CHOPN přes signifikantní zmírnění bronchiální obstrukce nedojde k poklesu parametrů oxidačního stresu (NO2-, NO3- a malondialdehydu).
The aim was to find out whether oxidative stress is reduced after resolution of even severe exacerbations requiring hospitalization. For the assessment, three substances pairticipating in oxidative stress were selected - nitrites (N02-) and nitrates (NO3-), i.e. substances produced by conversion from nitric oxide (NO) and malondialdehyde, a product of lipid peroxidation. Concentrations of NO2- and NO3- were measured in exhaled breath condensate (EBC) collected using the EcoScreen device (VIASYS HealthCare GmbH, Hoechberg, Germany). The group comprised 29 persons, 18 (62.1 %) males and 11 (37.9 %) females hospitalized for a COPD exacerbation. The median concentrations on admission and at discharge were as follows: 5.11 and 6.58 for NO2- (μmol/11, 42.3 and 53.0 for NO3- (μmol/l) and 30.5 and 17.7 for malondialdehyde (nmol/1), respectively. In none of the studied parameters, a significant decrease was observed. The results suggest that despite significeint resolution of bronchial obstruction in patients hospitalized for a COPD exacerbation, oxidative stress parameters (NO2- , N03- and malondialdehyde) are not reduced.
