Debridement je jedním z nejdůležitějších kroků k přípravě rány před chirurgickým úzávěrem, který může být proveden v různých formách a různými nástroji. Cíl: Cílem prospektivní případové studie bylo porovnat vliv dvou různých typů debridementu (nekrektomie) na parametry oxidativního stresu. Soubor a metodika: Do studie bylo zařazeno celkem pět pacientů s hlubokými dekubity v různých lokalizacích. Dekubitus byl rozdělen na dvě poloviny; jedna polovina rány byla ošetřena ostrou nekrektomií a druhá Versajet® hydrosystémem. Vzorky tkání, krve a moči byly odebrány v nultý a sedmý den po debridementu. Byla provedena histopatologická analýza a vyšetření parametrů oxidativního stresu v plazmě a moči. Výsledky: Byly nalezeny rozdíly v kvalitě a kvantitě granulační tkáně mezi dvěma různými typy provedeného debridementu. Zjištěno bylo nesignifikantní snížení markerů oxidativního stresu v plazmě a moči sedmý den po debridementu. Závěr: Předběžné a pilotní výsledky naznačují, že proces hojení ran je úzce spojen s markery oxidativního stresu, které jsou měřitelné v krevní plazmě i v moči. Tyto by mohly poukazovat na průběh hojení. U všech parametrů oxidativního stresu bylo pozorováno nesignifikantní snížení sedmý den po zákroku. Na pilotní studii naváže podrobná molekulárně-biologická analýza vzorků tkáně.
Wound debridement is one of the crucial steps in wound bed preparation for surgical closure. Various debridement forms and tools can be employed. Aim: The prospective case series study aimed to compare the impact of two different types of debridement on oxidative stress parameters. Material and methods: This study included five patients with pressure ulcers of deep category various localisation. The wound was divided into halves. In one, the sharp debridement was performed, in the second Hydrosurgery Versajet® debridement was accomplished. Tissue, blood and urine samples were collected on days 0 and 7 after the surgery. Histopathological evaluation in tissue samples was performed. Oxidative stress parameters in plasma and urine were evaluated. Results: Differences in quality and quantity of granulation tissue between two types of debridement were found. An insignificant decrease of oxidative stress markers in blood plasma and urine 7 days after surgery were observed. Conclusions: Preliminary and pilot results suggest that the wound healing process is closely associated with markers of oxidative stress that are measurable in blood plasma and urine. These could be indicative of the healing process. A nonsignificant decrease was observed for all oxidative stress parameters on day 7 after the surgery. The pilot study will be followed by a detailed molecular biological analysis of tissue samples.
- MeSH
- debridement MeSH
- dekubity * patofyziologie terapie MeSH
- klinická studie jako téma MeSH
- lidé MeSH
- oxidační stres MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- práce podpořená grantem MeSH
- MeSH
- amputace MeSH
- chirurgické laloky * MeSH
- dolní končetina patologie MeSH
- hojení ran MeSH
- infekce v ráně * terapie MeSH
- lidé středního věku MeSH
- lidé MeSH
- výsledek terapie MeSH
- zákroky plastické chirurgie * metody MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Heparinem indukovaná trombocytopenie (HIT) je imunologicky zprostředkovaná komplikace navazující zpravidla na expozici heparinu, méně často se rozvíjí po expozici jiným léčivům anebo spontánně. Mezi rizikové faktory rozvoje HIT patří typ použitého heparinu, jeho dávka a způsob aplikace i doba expozice, velké trauma či operace a obezita. Pravděpodobnost HIT dobře koreluje s tzv. 4T‑skóre. Laboratorní testy k potvrzení či vyloučení HIT se mají provádět u pacientů se střední až vysokou pravděpodobností HIT (4T‑skóre více než 3 body). Screeningové testy jsou založeny na průkazu protilátek proti komplexům heparinu a destičkového faktoru 4, cílem konfirmačních testů je průkaz aktivace trombocytů. Léčba HIT spočívá v okamžitém ukončení aplikace heparinu a v důsledné antitrombotické léčbě alternativními modalitami. Autoři popisují případ HIT u interně polymorbidní obézní pacientky s rozsáhlou infikovanou popáleninou projevující se extrémní urtikariální reakcí v místě aplikace nadroparinu a trombotizací žil podkoží. Díky včasné diagnostice a léčbě fondaparinuxem nedošlo k rozvoji závažnějších tromboembolických příhod.
Heparin‑induced thrombocytopenia (HIT) is an immunologically‑mediated complication, which usually follows heparin exposition, less frequently exposition to other drugs or even occurs spontaneously. The type of heparin, its dose and mode of application as well as the exposition time, major trauma or operation, and obesity represent the main risk factors for HIT. The probability of HIT correlates with so‑called 4T‑score. A confirmatory laboratory diagnostic should be exclusively reserved for patients with a medium to a high probability of HIT development (more than 3 points in 4T‑score). The screening method is based on serological detection of antibodies against heparin‑platelet factor‑4 complexes; confirmation tests aim to identify the activation of platelets. The treatment of HIT requires an immediate interruption of heparin application and rigorous antithrombotic treatment with an alternative agent. Herein authors describe a clinical case of HIT manifested as an extreme urticarial reaction in the location of nadroparin application as well as thrombosis of deep subcutaneous veins in a polymorbid obese patient with an extensive and infected burn. Due to timely diagnosis and fondaparinux treatment, no more severe thrombotic events occurred in this patient.
- Klíčová slova
- 4 T‑skóre,
- MeSH
- antikoagulancia terapeutické užití MeSH
- fondaparinux terapeutické užití MeSH
- heparin * škodlivé účinky MeSH
- lidé středního věku MeSH
- lidé MeSH
- trombocytopenie * chemicky indukované MeSH
- trombóza farmakoterapie MeSH
- výsledek terapie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
Pancreatic α-amylase plays an important role in dietary starch hydrolysis in the small intestine and participates in enhanced glucose concentration after meals. It seems to be a problem for diabetic patients, who suffer from longer postprandial hyperglycemia after meal consumption than healthy people. There are commercially available drugs that inhibit α-amylase and thus reduce the postprandial hyperglycemia effect. However, these drugs may cause severe side effects. Conversely, some naturally occurring flavonoids were suggested to have an α-amylase-inhibiting effect without any side effects. There had been no rapid, undemanding method in terms of sample and reagent preparation that would enable screening of many potential inhibitors. Therefore, we developed an online capillary electrophoresis method to monitor α-amylase activity in the presence of an inhibitor. Each reaction constituent was introduced separately, directly into a capillary where the reagents were mixed by diffusion, which resulted in a 5-min analysis including conditioning of the capillary. We applied the method to test the inhibitory effect of flavonoid standards and their mixture and we investigated the inhibitory effect of ethanolic extract from Betula pendula bark. The developed method presents a faster and less expensive alternative to previously described offline methods. Graphical abstract Online CE screening of α-amylase inhibitors.
Dasatinib is a novel oral prescription drug proposed for treating adult patients with chronic myeloid leukemia. Three analytical methods, namely ultra high performance liquid chromatography, capillary zone electrophoresis, and sequential injection analysis, were developed, validated, and compared for determination of the drug in the tablet dosage form. The total analysis time of optimized ultra high performance liquid chromatography and capillary zone electrophoresis methods was 2.0 and 2.2 min, respectively. Direct ultraviolet detection with detection wavelength of 322 nm was employed in both cases. The optimized sequential injection analysis method was based on spectrophotometric detection of dasatinib after a simple colorimetric reaction with folin ciocalteau reagent forming a blue-colored complex with an absorbance maximum at 745 nm. The total analysis time was 2.5 min. The ultra high performance liquid chromatography method provided the lowest detection and quantitation limits and the most precise and accurate results. All three newly developed methods were demonstrated to be specific, linear, sensitive, precise, and accurate, providing results satisfactorily meeting the requirements of the pharmaceutical industry, and can be employed for the routine determination of the active pharmaceutical ingredient in the tablet dosage form.
- MeSH
- chemie farmaceutická metody normy MeSH
- dasatinib analýza MeSH
- elektroforéza kapilární * MeSH
- lidé MeSH
- limita detekce MeSH
- reprodukovatelnost výsledků MeSH
- spektrofotometrie * MeSH
- tablety chemie normy MeSH
- vysokoúčinná kapalinová chromatografie * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
High anion gap metabolic acidosis frequently complicates acute paracetamol overdose and is generally attributed to lactic acidosis or compromised hepatic function. However, metabolic acidosis can also be caused by organic acid 5-oxoproline (pyroglutamic acid). Paracetamol's toxic intermediate, N-acetyl-p-benzoquinoneimine irreversibly binds to glutathione and its depletion leads to subsequent disruption of the gamma glutamyl cycle and an excessive 5-oxoproline generation. This is undoubtedly an underdiagnosed condition because measurement of serum 5-oxoproline level is not readily available. A simple, cost effective, and fast capillary electrophoresis method with diode array detection (DAD) for simultaneous measurement of both paracetamol (acetaminophen) and 5-oxoproline in serum was developed and validated. This method is highly suitable for clinical toxicology laboratory diagnostic, allowing rapid quantification of acidosis inducing organic acid 5-oxoproline present in cases of paracetamol overdose. The calibration dependence of the method was proved to be linear in the range of 1.3-250μgmL-1, with adequate accuracy (96.4-107.8%) and precision (12.3%). LOQ equaled 1.3μgmL-1for paracetamol and 4.9μgmL-1for 5-oxoproline.
- MeSH
- acidóza MeSH
- elektroforéza kapilární * MeSH
- kyselina pyrrolidonkarboxylová MeSH
- lidé MeSH
- neopioidní analgetika MeSH
- paracetamol MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
In this study, we optimized a method for the determination of free amino acids in Nicotiana tabacum leaves. Capillary electrophoresis with contactless conductivity detector was used for the separation of 20 proteinogenic amino acids in acidic background electrolyte. Subsequently, the conditions of extraction with HCl were optimized for the highest extraction yield of the amino acids because sample treatment of plant materials brings some specific challenges. Central composite face-centered design with fractional factorial design was used in order to evaluate the significance of selected factors (HCl volume, HCl concentration, sonication, shaking) on the extraction process. In addition, the composite design helped us to find the optimal values for each factor using the response surface method. The limits of detection and limits of quantification for the 20 proteinogenic amino acids were found to be in the order of 10-5and 10-4 mol l-1, respectively. Addition of acetonitrile to the sample was tested as a method commonly used to decrease limits of detection. Ambiguous results of this experiment pointed out some features of plant extract samples, which often required specific approaches. Suitability of the method for metabolomic studies was tested by analysis of a real sample, in which all amino acids, except for L-methionine and L-cysteine, were successfully detected. The optimized extraction process together with the capillary electrophoresis method can be used for the determination of proteinogenic amino acids in plant materials. The resulting inexpensive, simple, and robust method is well suited for various metabolomic studies in plants. As such, the method represents a valuable tool for research and practical application in the fields of biology, biochemistry, and agriculture.
Fluorides and fluoroaluminates decrease mouse sperm fertilizing potential by modifying the process of sperm preparation for fertilization, so-called capacitation, followed by acrosome reaction (AR). Capacitation was monitored by protein tyrosine phosphorylation (pTyr), and AR was induced consequently. The aim of this study was to apply kinetic analysis to the previously obtained dependences of pTyr and AR at capacitation times, and propose a mathematical theory for a mechanism when sperm maturation ability is amended by external stimuli. The experimental input data, previously obtained, are consistent with the proposed theory and the results of kinetic analysis show that sperm capacitation runs as two subsequent first-order steps. Firstly, an unstable intermediate is formed and then gradually decomposes. The time corresponding to the maximal production of the unstable intermediate is probably most suitable for sperm obtaining the ability to fertilize the egg. The presented calculations indicate that the application of kinetic analysis can serve as a tool to predict or confirm a course of biological events that are modified by external factors, and therefore the proposed theory shall be of interest to a broad scientific audience.
- MeSH
- akrozomální reakce * MeSH
- fluor škodlivé účinky farmakologie MeSH
- fluoridy škodlivé účinky farmakologie MeSH
- hliník škodlivé účinky farmakologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- spermie cytologie účinky léků fyziologie MeSH
- zrání spermie MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
