The study aim was to compare molecular-level effects (blood-dialyzer interactions) of heparin and citrate anticoagulation using proteome-wide analysis of biofilm adsorbed to dialysis membrane. Ten patients receiving maintenance hemodialysis were examined in a crossover design under three different anticoagulation regimens, namely citrate, heparin, and anticoagulation-free (control). Following a regular hemodialysis session (4 hours, polysulfone membrane), dialyzers were flushed and the surface biofilm eluted by acetic acid. Protein composition of the eluates was determined by 2-dimensional gel electrophoresis and resulting patterns compared between regimens. Proteins responsible for the difference were identified by mass spectrometry. Citrate anticoagulation was associated with significantly less protein adsorption to the membrane than heparin (2.2 [1.1-2.9] mg vs. 6.5 [2.9-11.6] mg, P = 0.009). Among the proteins identified as major discriminators between citrate and the other regimens, fibrin α-chain fragments of molecular weight below 40 kDa prevailed. In these fragments, an analysis of the amino acid sequence has been performed by comparison with the UniProt database. It showed missing α-chain cross-links. On the contrary, heparin prevented adsorption and cleavage of several heparin-binding proteins; especially complement factor H-related protein 3, insulin-like growth factor binding proteins (2, 4, and 5), and chemerin. Compared to heparin, citrate is associated with less protein adsorption and imperfectly crosslinked fibrin clot formation. Membrane adsorptive properties are significantly modified by the anticoagulation regimen.
- MeSH
- adsorpce MeSH
- antikoagulancia farmakologie MeSH
- dialýza ledvin přístrojové vybavení metody MeSH
- hemokoagulace účinky léků MeSH
- heparin farmakologie MeSH
- krevní proteiny analýza MeSH
- kyselina citronová farmakologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- membrány umělé MeSH
- polymery chemie MeSH
- proteomika MeSH
- senioři MeSH
- sulfony chemie MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
Indoxyl sulfate has been identified as a major factor in the dysregulation of several genes. It is classified as a poorly dialyzable uremic toxin and thus a leading cause in the poor survival rate of dialysis patients. A monocentric, prospective, open cohort study was performed in 43 male patients undergoing chronic renal replacement therapy in a single hemodialysis center. The aim of the study was to determine the influence of acetate- versus citrate-buffered dialysis fluids in hemodialysis (HD) and postdilution hemodiafiltration (HDF) settings on the elimination of indoxyl sulfate. Also, additional factors potentially influencing the serum concentration of indoxyl sulfate were evaluated. For this purpose, the predialysis and postdialysis concentration ratio of indoxyl sulfate and total protein was determined. The difference was of 1.15 (0.61; 2.10), 0.89 (0.53; 1.66), 0.32 (0.07; 0.63), and 0.44 (0.27; 0.77) μmol/g in acetate HD and HDF and citrate HD and HDF, respectively. Acetate HD and HDF were superior when concerning IS elimination when compared to citrate HD and HDF. Moreover, residual diuresis was determined as the only predictor of lower indoxyl sulfate concentration, suggesting that it should be preserved as long as possible. This trial is registered with EU PAS Register of Studies EUPAS23714.
- MeSH
- acetáty farmakologie MeSH
- dialýza ledvin metody MeSH
- dialyzační roztoky chemie farmakologie MeSH
- hemodiafiltrace metody MeSH
- hydrogenuhličitany MeSH
- indican krev farmakokinetika MeSH
- kyselina citronová krev farmakologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- nemoci ledvin terapie MeSH
- prospektivní studie MeSH
- senioři MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- Publikační typ
- časopisecké články MeSH
- klinické zkoušky MeSH
- Klíčová slova
- Picoprep,
- MeSH
- fixní kombinace léků MeSH
- klinické zkoušky, fáze III jako téma MeSH
- kyselina citronová farmakologie terapeutické užití MeSH
- lidé MeSH
- oxid hořečnatý farmakologie terapeutické užití MeSH
- předoperační péče metody MeSH
- projímadla * farmakologie terapeutické užití MeSH
- randomizované kontrolované studie jako téma MeSH
- spokojenost pacientů MeSH
- Check Tag
- lidé MeSH
- MeSH
- akutní poškození ledvin * diagnóza etiologie terapie MeSH
- časové faktory MeSH
- dialýza ledvin metody MeSH
- hemodialyzační roztoky MeSH
- kyselina citronová farmakologie terapeutické užití MeSH
- lidé MeSH
- péče o pacienty v kritickém stavu metody MeSH
- vápníkové chelátory terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- novinové články MeSH
Brominated furanone and epigallocatechin gallate (EGCG) are naturally occurring polyphenolic compounds that can be derived from sources such as Delisea pulchra algae and green tea, respectively. These compounds may have potential health benefits and antimicrobial properties. Biofilm formation and bacterial motility are virulence factors that seem to be involved in the autoinducer 2 (AI-2)-mediated quorum sensing (QS) response of Campylobacter. In this study, the anti-QS activities of 2(5H)-furanone, EGCG, and a citric-based disinfectant were tested against Campylobacter jejuni. The minimal bactericidal concentration (MBC) was determined by a microdilution method, and the AI-2 activity was measured by bioluminescence. For motility tests, subinhibitory concentrations of each compound were mixed with semisolid Muller Hinton agar. Biofilm formation was quantified in broth-containing microplates after staining with safranin. The MBC of tested compounds ranged from 0.3 to 310 μg/mL. Subinhibitory concentrations of all of the antimicrobial compounds significantly decreased (19 to 62 %) the bacterial motility and reduced biofilm formation. After treatment with EGCG, furanone, and the disinfectant, AI-2 activity was decreased by 60 to 99 % compared to control. In conclusion, 2(5H)-furanone, EGCG, and the disinfectant exert bactericidal effects against C. jejuni and disturb QS activity and reduce motility and biofilm formation. These compounds may be naturally occurring alternatives to control C. jejuni.
- MeSH
- biofilmy účinky léků MeSH
- Campylobacter jejuni účinky léků genetika fyziologie MeSH
- dezinficiencia farmakologie MeSH
- gama-butyrolakton analogy a deriváty farmakologie MeSH
- katechin analogy a deriváty farmakologie MeSH
- kyselina citronová farmakologie MeSH
- mikrobiální testy citlivosti MeSH
- quorum sensing účinky léků MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
BACKGROUND: Flow cytometry represents an attractive approach for developing currently unavailable screening tests for prion diseases. Several studies have reported significant differences in the binding of antibodies directed against cellular prion protein (PrP(C)) to blood cells of prion-infected subjects compared with healthy controls. However, flow cytometry data usually show large individual variations in detected PrP(C) levels in both infected and control groups, rendering the interpretation of individual patient data difficult. OBJECTIVES: To determine how pre-analytical variables, such as the choice of anticoagulant, whether or not the blood was stored, and the storage temperature, affect the detection of PrP(C) in blood cells. METHODS: Blood from healthy donors was collected in EDTA or citrate anticoagulant and processed either immediately or after storage overnight at room temperature or at 4°C. The expression of PrP(C) by T cells, B cells, NK cells, monocytes and circulating dendritic cells was evaluated using quantitative flow cytometry with the PrP(C) monoclonal antibodies AG4 and AH6. RESULTS: The anticoagulation of blood with citrate resulted in decreased levels of PrP(C) on monocytes but not the other cell types. The storage of blood prior to analysis led to a significant decrease in the levels of PrP(C) on the cells studied, although there were substantial differences between the cell populations. This decrease was more pronounced when using mAb AG4, which targets the N-terminal portion of the PrP(C) molecule, or following storage at room temperature. Moreover, we identified platelet satellitism on leukocytes, especially on monocytes and granulocytes, as an additional factor contributing to the heterogeneity of PrP(C) detection in stored blood. CONCLUSIONS: Our study demonstrates that the storage of blood prior to analysis greatly affects the detection of PrP(C) by flow cytometry. To limit the inclusion of storage-generated artifacts, we recommend the processing of blood samples immediately after their collection.
- MeSH
- antikoagulancia farmakologie MeSH
- dendritické buňky chemie MeSH
- dospělí MeSH
- EDTA farmakologie MeSH
- kyselina citronová farmakologie MeSH
- leukocyty chemie MeSH
- lidé středního věku MeSH
- lidé MeSH
- odběr vzorku krve metody MeSH
- priony analýza MeSH
- průtoková cytometrie metody MeSH
- trombocyty MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Cíl: Zjistit vliv pomocných látek edetanu disodného dihydrátu a monohydrátu kyseliny citronové na rychlost rozpouštění antibiotického prášku, na velikost částic po naředění a na chemickou stabilitu přípravku po naředění vodou na injekci a 5% roztokem glukózy. Materiál a metody:Porovnával se přípravek obsahující pomocné látky (Tazocin) s generickými přípravky, které pomocné látky neobsahují. Rychlost rozpouštění se stanovovala organolepticky a spektrofotometricky, velikost částic se hodnotila mikroskopicky po naředění a po 4 hodinách stání při laboratorní teplotě a chemická stabilita metodou HPLC. Výsledky: Tazocin se v obou zkoumaných rozpouštědlech výrazně rychleji rozpouštěl ve srovnání s přípravky bez pomocných látek (3–4 minuty oproti 10–13 minutám). Limit pro maximální počet částic větších než 25 mm v naředěném přípravku nebyl překročen u žádného přípravku ani po naředění vodou na injekci ani po naředění 5% roztokem glukózy, a to ani hned po naředění, ani po 4 hodinách stání při laboratorní teplotě. Nicméně pouze u Tazocinu se počet nežádoucích velkých částic s časem stání naředěného přípravku zmenšoval v obou rozpouštědlech. Edetan disodný dihydrát a monohydrát kyseliny citronové stabilizovaly rozpuštěné částice léčivé látky a zabraňovaly přechodu léčivé látky na nerozpustnou formu, což se projevovalo snižujícím se počtem nežádoucích velkých částic. Tazocin byl stabilnější, stabilita však nebyla statisticky významně lepší v porovnání s jeho generiky. Závěr: Edetan disodný dihydrát a monohydrát kyseliny citronové (případně i použitá technologie lyofilizace) tak mají pozitivní vliv na přípravu a vlastnosti antibiotického injekčního roztoku.
Objective: To evaluate the effect of excipients disodium-edetate dihydrate and citric acid monohydrate on rate of antibiotic powder dissolution, particle size after dissolution and chemical stability of the product after reconstitution with water for injection and 5% glucose solution. Material and methods: The product containing excipients (Tazocin) was compared with generic products without excipients. Rate of dissolution was evaluated organoleptically and spectrophotometrically, particle size was determined microscopically after reconstitution and after 4 hours of standing at laboratory temperature and chemical stability was assessed by HPLC. Results: Tazocin dissolved significantly faster in both solvents compared to products without excipients (3–4 min versus 10–13 min). The limit for maximal number of particles larger than 25 mm in the reconstituted solution was not exceeded in any of the products either after using water for injection or after using 5% glucose, both after reconstitution and after 4 hours of standing at laboratory temperature. Nevertheless, only for Tazocin the number of unwanted larger particles decreased in time in both solvents. Disodium-edetate dihydrate and citric acid stabilized the dissolved particles of the active substance and prevented its transformation into the insoluble form, which results in lower number of unwanted large particles. Tazocin was more stable, but the stability was not significantly better than its generic products. Conclusions: Disodium-edetate dihydrate and citric acid monohydrate (and possibly the used method of lyophilisation) therefore have a positive effect on the preparation and properties of antibiotic solution for intravenous application.
- Klíčová slova
- chemická stabilita, rychlost rozpouštění, Tazocin,
- MeSH
- antibakteriální látky aplikace a dávkování chemie MeSH
- EDTA farmakologie MeSH
- injekce intravenózní MeSH
- intravenózní infuze MeSH
- kyselina citronová farmakologie MeSH
- kyselina penicilanová analogy a deriváty aplikace a dávkování chemie MeSH
- piperacilin MeSH
- pomocné látky farmakologie MeSH
- rozpustnost MeSH
- velikost částic MeSH
- Publikační typ
- srovnávací studie MeSH
Swellable-matrix tablets based on hydrophilic polymers are simple dosage forms with controlled release, which are currently used in pharmacotherapy of many diseases. The most commonly used polymer is (hydroxypropyl)methylcellulose (hypromelose). The drug release from these systems through a gel layer can be modified using additives changing the drug dissolution profile. These excipients can be divided into several groups such as indifferent fillers (soluble or insoluble), substances increasing drug solubility (ß-cyclodextrins), excipients forming interactive products (polymers, ionexchange resins, surfactants) and substances influencing pH in matrix microenvironment (acid and basic modifiers).
- MeSH
- aplikace orální MeSH
- cyklodextriny farmakokinetika farmakologie terapeutické užití MeSH
- farmaceutické pomocné látky farmakokinetika farmakologie terapeutické užití MeSH
- fumaráty farmakokinetika farmakologie terapeutické užití MeSH
- gely farmakokinetika farmakologie terapeutické užití MeSH
- kyselina askorbová farmakokinetika farmakologie terapeutické užití MeSH
- kyselina citronová farmakokinetika farmakologie terapeutické užití MeSH
- lékové formy MeSH
- lékové interakce MeSH
- léky s prodlouženým účinkem farmakokinetika farmakologie MeSH
- lidé MeSH
- methylcelulosa farmakokinetika farmakologie terapeutické užití MeSH
- nosiče léků farmakokinetika farmakologie MeSH
- Check Tag
- lidé MeSH
- MeSH
- antitusika farmakokinetika farmakologie MeSH
- aplikace orální MeSH
- experimenty na zvířatech MeSH
- financování organizované MeSH
- kodein farmakologie MeSH
- kyselina citronová farmakologie MeSH
- léčivé rostliny účinky léků MeSH
- Meliaceae chemie MeSH
- morčata MeSH
- nemoci dýchací soustavy farmakoterapie MeSH
- pletysmografie metody využití MeSH
- polysacharidy farmakokinetika farmakologie MeSH
- zvířata MeSH
- Check Tag
- morčata MeSH
- zvířata MeSH
- Publikační typ
- abstrakty MeSH
