buffering ability
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In this work, two mixed-mode columns from a different manufacturers and one marketed as a reversed-phase column were characterized and compared in the terms of their interaction abilities, retentivity, peak symmetry, and applicability for peptide separation. All the tested columns contain octadecyl ligand and positively charged modifier, i.e. pyridyl group for the reversed-phase column XSelect CSH C18, quaternary alkylamine for mixed-mode column Atlantis PREMIER BEH C18 AX, and permanently charged moiety (details not available from the manufacturer) for mixed-mode column Luna Omega PS C18. For detailed characterization and comparison of their interaction potential, several approaches were used. First, a simple Walters test was performed to estimate hydrophobic and silanophilic interactions of the tested columns. The highest values of both parameters were observed for column Atlantis PREMIER BEH C18 AX. To investigate the effect of pH and buffer concentration on retention, mobile phases composed of acetonitrile and buffer (ammonium formate, pH 3.0; ammonium acetate pH 4.7 and pH 6.9) in various concentrations (5mM; 10mM; 15mM and 20mM) were used. The analysis of permanently charged compounds was used to describe the electrostatic interaction abilities of the stationary phases. The most significant contribution of electrostatic interactions to the retention was observed for Atlantis PREMIER BEH C18 AX column in the mobile phase with buffer of pH 3.0. A set of ten dipeptides, three pentapeptides and one octapeptide was used to investigate the effects of pH and buffer concentration on retention and peak symmetry. Each of the tested columns provides the optimal peak shape under different buffer pH and concentration. The gradient separation of the 14 tested peptides was used to verify the application potential of the tested columns for peptide separation. The best separation was achieved within 4 minutes on column Atlantis PREMIER BEH C18 AX.
We present a new theoretical approach for calculating changes in the physico-chemical properties of BGEs for measurements by CZE due to the electrolysis in electrode vials (vessels). Electrolysis is an inevitable phenomenon in any measurement in CZE. Water electrolysis, which occurs in most measurements, can significantly alter the composition of the BGE in electrode vials and in the separation capillary and has a negative influence on the robustness and quality of separations. The ability to predict changes in the composition of the BGE is important for evaluation of the suitability of the BGEs for repeating electrophoretic runs. We compared theoretically calculated changes in the physico-chemical properties (pH, conductivity) with those measured using pH-microelectrode and contactless conductivity detection of the BGE after the electrophoretic run. We confirmed the validity of our theoretical approach with a common BGE composed of acid-base pair, where one constituent is fully dissociated while the second constituent is dissociated by only half, and with Good's buffer. As predicted by theoretical approach, the changes in the physico-chemical properties of the Good's buffer after the electrophoretic run were several times lower than in the case of a common BGE composed of a weak acid - strong base pair.
An improved preparative method based on isoelectric focusing of analytes in a cellulose-based separation medium is described in this study. Cellulose is suspended in an aqueous solution of simple buffers, ethylene glycol, glycerol, nonionic surfactant, and colored pI markers. Water partially evaporates during focusing run and the separation takes place in an in situ generated layer of cellulose, which has a gel-like appearance at the end of analysis. Final positions of analytes are indicated by the positions of zones of focused pI markers. Fractions, segments of the separation medium with analytes, can be simply collected by spatula and analyzed by downstream analytical methods. Good focusing ability of the new method and almost quantitative recovery of model proteins, cytochrome c and bovine serum albumin, was verified by gel electrophoresis and capillary isoelectric focusing of the collected fractions.
The aim of this study was to develop novel hydrogel-based beads and characterize their potential to deliver and release a drug exhibiting pH-dependent solubility into distal parts of gastrointestinal (GI) tract. Oxycellulose beads containing diclofenac sodium as a model drug were prepared by the ionotropic external gelation technique using calcium chloride solution as the cross-linking medium. Resulting beads were characterized in terms of particle shape and size, encapsulation efficacy, swelling ability and in vitro drug release. Also, potential drug-polymer interactions were evaluated using Fourier transform infrared spectroscopy. The particle size was found to be 0.92-0.96 mm for inactive (oxycellulose only) and 1.47-1.60 mm for active (oxycellulose-diclofenac sodium) beads, respectively. In all cases, the sphericity factor was between 0.70 and 0.81 with higher values observed for samples containing higher polymer and drug concentrations. The swelling of inactive beads was found to be strongly influenced by the pH and composition (i.e. Na(+) concentration) of the selected media (simulated gastric fluid vs. phosphate buffer pH 6.8). The encapsulation efficiency of the prepared particles ranged from 58% to 65%. Results of dissolution tests showed that the drug loading inside of the particles influenced the rate of its release. In general, prepared particles were able to release the drug within 12-16 h after a lag time of 4 h. Fickian diffusion was found as the predominant drug release mechanism. Thus, this novel particulate system showed a good potential to deliver drugs specifically to the distal parts of the human GI tract.
- MeSH
- antiflogistika nesteroidní chemie MeSH
- chemické modely MeSH
- difuze MeSH
- diklofenak chemie MeSH
- farmaceutická chemie MeSH
- farmaceutická technologie metody MeSH
- kinetika MeSH
- koncentrace vodíkových iontů MeSH
- léky s prodlouženým účinkem MeSH
- methylcelulosa analogy a deriváty chemie MeSH
- nosiče léků MeSH
- příprava léků MeSH
- pufry MeSH
- rozpustnost MeSH
- spektroskopie infračervená s Fourierovou transformací MeSH
- studie proveditelnosti MeSH
- velikost částic MeSH
- žaludeční šťáva chemie MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Thermoregulatory behaviour represents an important component of ectotherm non-genetic adaptive capacity that mitigates the impact of ongoing climate change. The buffering role of behavioural thermoregulation has been attributed solely to the ability to maintain near optimal body temperature for sufficiently extended periods under altered thermal conditions. The widespread occurrence of plastic modification of target temperatures that an ectotherm aims to achieve (preferred body temperatures) has been largely overlooked. I argue that plasticity of target temperatures may significantly contribute to an ectotherm's adaptive capacity. Its contribution to population persistence depends on both the effectiveness of acute thermoregulatory adjustments (reactivity) in buffering selection pressures in a changing thermal environment, and the total costs of thermoregulation (i.e. reactivity and plasticity) in a given environment. The direction and magnitude of plastic shifts in preferred body temperatures can be incorporated into mechanistic models, to improve predictions of the impact of global climate change on ectotherm populations.
- MeSH
- aklimatizace MeSH
- chování zvířat MeSH
- klimatické změny MeSH
- selekce (genetika) MeSH
- teplota MeSH
- termoregulace MeSH
- životní prostředí MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The release of water from storage compartments to the transpiration stream is an important functional mechanism that provides the buffering of sudden fluctuations in water potential. The ability of tissues to release water per change in water potential, referred to as hydraulic capacitance, is assumed to be associated with the anatomy of storage tissues. However, information about how specific anatomical parameters determine capacitance is limited. In this study, we measured sapwood capacitance (C) in terminal branches and roots of five temperate tree species (Fagus sylvatica L., Picea abies L., Quercus robur L., Robinia pseudoacacia L., Tilia cordata Mill.). Capacitance was calculated separately for water released mainly from capillary (CI; open vessels, tracheids, fibres, intercellular spaces and cracks) and elastic storage compartments (CII; living parenchyma cells), corresponding to two distinct phases of the moisture release curve. We found that C was generally higher in roots than branches, with CI being 3-11 times higher than CII Sapwood density and the ratio of dead to living xylem cells were most closely correlated with C In addition, the magnitude of CI was strongly correlated with fibre/tracheid lumen area, whereas CII was highly dependent on the thickness of axial parenchyma cell walls. Our results indicate that water released from capillary compartments predominates over water released from elastic storage in both branches and roots, suggesting the limited importance of parenchyma cells for water storage in juvenile xylem of temperate tree species. Contrary to intact organs, water released from open conduits in our small wood samples significantly increased CI at relatively high water potentials. Linking anatomical parameters with the hydraulic capacitance of a tissue contributes to a better understanding of water release mechanisms and their implications for plant hydraulics.
- MeSH
- dřevo anatomie a histologie MeSH
- stromy anatomie a histologie MeSH
- voda metabolismus MeSH
- xylém metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
Úvod: Suplementace ß-alaninem se stává běžnou praxí u vrcholových sportovců a také se stává objektem zkoumání odborníků, především fyziologů a sportovních lékařů na celém světě. Doplněním ß-alaninu dochází k vzestupu koncentrace karnosinu ve svalových buňkách, což vede k oddálení svalové únavy a ke snazší regeneraci při opakovaných sportovních výkonech vysoké intenzity. Karnosin je dipeptid skládající se z beta-alaninu a L-histidinu. ß-alanin má největší účinnost u výkonů, které jsou silně závislé na syntéze ATP z anaerobní glykolýzy. Cíl: Ze zkušenosti víme, že výrobci a distributoři potravních doplňků mají sklon účinky svých produktů silně nadhodnocovat. Cílem práce je shrnout a objektivně zhodnotit efekt suplementace ß-alaninu na sportovní výkon na základě nejnovějších poznatků světového písemnictví. Metodika: Použili jsme čtyři databáze odborných prací Google Scholar, PubMed, Sport- Discus a Scopus. Na základě klíčových slov jsme vyhledávali práce od roku 1990 do současnosti. Vybírali jsme jen dvojitě zaslepené studie s kontrolní skupinou a placebem. Výsledky: Nalezli jsme celkem 114 prací požadované kvality a 8 souborných článků typu review. Významné ergogenní účinky byly zjištěny především u netrénovaných jedinců při zátěžových testech v laboratorních podmínkách. Především došlo k zlepšení krátkodobých vytrvalostních výkonů v době trvání 60–240 s. Hlavní mechanismus účinků je připisován oddálení únavy díky intracelulárním pufrovacím schopnostem karnosinu, které jsou nezávislé na bikarbonátovém systému. U dobře trénovaných sportovců je ergogenní efekt ß-alaninu menší. Vytrvalostní schopnosti se však nezvýšily. Doporučená denní dávka (DDD) ß-alaninu je 1,6–6,4 g denně. Jediným nežádoucím účinkem ß-alaninu mohou být parestesie. Proto se doporučuje rozdělit DDD do více dávek. Závěry: Poslední odborné práce ukazují, že u trénovaných sportovců může ß-alanin zvýšit sportovní výkon především nepřímo: zvýšením anaerobního prahu (AT), zvýšením VO2 na úrovni AT, oddálením svalové únavy, schopností zvýšit tréninkový objem či prodloužením doby do vyčerpání; vytrvalostní výkon se tak může zlepšit bez navýšení maximální aerobní kapacity. ß-alanin zlepšuje při vytrvalostních výkonech spurterské schopnosti. Některé účinky ß-alaninu na sportovní výkon přesto zůstávají i nadále sporné. Příčinu autoři nespatřují v rozdílné metodice, ale ve značné individuální responsibilitě lidského organizmu na suplementaci tímto doplňkem stravy.
Introduction: Supplementation with beta-alanine becomes not only a common practice among elite athletes, but also got into the spotlight of experts, especially physiologists and sports physicians worldwide. Adding of ß-alanine leads to increase in the concentration of carnosine in muscle cells, to delay of muscle fatigue, and to facilitatation of recovery during repeated sports performances of high intensity. ß-alanine is most efficient for the performances, which are heavily dependent on the synthesis of ATP from anaerobic glycolysis. Carnosine is a dipeptide consisting of beta-alanine and L-histidine. Objective: We know from experience that manufacturers and distributors of dietary supplements tend effects of their products greatly overestimate. Our aim is to summarize and objectively evaluate the effect of ß-alanine supplementation on athletic performance based on the latest knowledge of world literature. Methods: We used four databases of scholarly works Google Scholar, Scopus, PubMed, and SPORTDiscus. The key words are searched for work from 1990 to the present. We chose only double-blind studies with a control group and placebo. Results: We found 114 high-quality work and 8 review articles. Significant ergogenic effects were generally found with untrained individuals during load tests under laboratory conditions. Above all, there was an improvement of short-persevere-speed performance for the duration of 60 to 240 s. The main mechanism of the effect is attributed to delay fatigue due to the buffering capabilities of carnosine which is independent off the bicarbonate buffering system. For welltrained athletes is ergogenic effect of carnosine smaller. Maximal aerobic power and VO2max did not increase. The recommended daily allowance (RDA) of beta-alanine is 1.6 to 6.4 g .day-1. The only side effect of carnosine can be paresthesia. Therefore it is recommended to divide the RDD into multiple doses a day. Conclusions: Recent scientific studies have shown that trained athletes can use ß-alanine to enhance athletic performance mainly indirectly: by increasing the anaerobic threshold (AT) or VO2AT, by delaying muscle fatigue, by improvement the ability to increase training volume or to prolong the time to exhaustion (TTE); thus, endurance performance can improve without increasing the maximum aerobic capacity. After endurance performance, carnosine improves the final sprint capabilities. Some effects of carnosine on athletic performance still remain controversial. Scientists do not see the cause of the difference in methodology, but in considerable individual responsibility of human organism to this supplement.
Diseases related to a disrupted skin barrier are accompanied by lower levels of ceramides in the stratum corneum (SC) lipid matrix. Delivering ceramides directly into damaged skin is a viable alternative to conventional corticosteroids, but is hindered by their low skin bioavailability and limited nanoformulation ability. Here, we developed stable liposomal systems containing ceramides and other SC lipids, and tested their effectiveness in skin barrier repair. Lipid film hydration and high-pressure homogenization were used to prepare different types of liposomes. To determine the stability, the particle size and polydispersity index were measured. The optimal systems were found to include ceramide 3 and 6, cholesterol and stearic acid, with 10% urea in phosphate-buffered saline as the aqueous phase. The ability of the system to repair chemically-damaged porcine skin was tested. While treatment by a standard lipid suspension reduced the passage of a model permeant only to a limited extent, drug flux through the liposomally-treated skin was much closer to permeation through intact skin. The non-homogenized liposomes were more effective than their homogenized version. These findings were also confirmed by FTIR measurements. This suggests that our approach to liposomal development has considerable potential for the repair of a disrupted skin barrier.
The surface plasmon resonance (SPR) biosensor system with dispersionless microfluidics for the direct and label-free detection of a soluble vascular endothelial growth factor receptor (sVEGFR-1) is described. The detection approach takes advantage of an affinity interaction between sVEGFR-1 and its ligand, vascular endothelial growth factor (VEGF-A), which is covalently immobilized on the surface of the SPR sensor. The ability of the immobilized VEGF-A to specifically bind the sVEGFR-1 receptor is demonstrated in a buffer. The detection of sVEGFR-1 in 2% human blood plasma is carried out by using the sequential injection approach. The detection limit of 25 ng/mL is achieved. In addition, we demonstrate that the functional surface of the sensor can be regenerated for repeated use.
- MeSH
- biologické markery krev MeSH
- biosenzitivní techniky metody MeSH
- lidé MeSH
- myelodysplastické syndromy krev diagnóza metabolismus MeSH
- povrchová plasmonová rezonance metody MeSH
- receptor 1 pro vaskulární endoteliální růstový faktor krev metabolismus MeSH
- vaskulární endoteliální růstový faktor A krev metabolismus MeSH
- vazba proteinů MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
Sarcosine has been identified as a potential prostate cancer marker. To provide determination of this compound, a number of methods are developing. In this study, we optimized a method for its separation by hydrophilic interaction LC with electrochemical detection (ED). Due to the fact that mobile phases commonly used for this type of separation altered the LODs measured by electrochemical detectors, we applied postcolumn dosing of buffer suitable for ED. The optimized conditions were mobile phase A acetonitrile, mobile phase B water in the ratio A/B 70:30, with postcolumn addition of mobile phase C (200 mM phosphate buffer pH 9). The optimal mixing ratio was A + B/C 1:1 with a flow rate of 0.80 mL/min (0.40 + 0.40 mL/min) and detection potential of 1000 mV. Due to the optimization of the parameters for effective separation, which had to meet the optimal parameters of ED, we reached a good resolution for separation also with a good LOD (100 nM). In addition, we successfully carried out sarcosine analysis bound on our modified paramagnetic microparticles with the ability to preconcentrate sarcosine isolated from artificial urine.
