In this work, we present the synthesis and application of fluorescent rhodamine B hydrazide for the derivatization of simple oligosaccharides and complex glycans using a hydrazone formation chemistry approach. The labeling conditions and the experimental setup of CE/LIF were optimized by analyzing oligosaccharide standards. The CE/LIF separations were performed in polybrene-coated capillaries eliminating the need for the purification step after derivatization. The addition of methanol to the background electrolyte significantly increased the LIF detection sensitivity reaching the limits of detection in the attomole range. The resolution of carbohydrate samples was improved by using long (98 cm) capillaries and polymer additives (polybrene). The developed method was applied for CE/LIF and CE-MS analysis of N-linked glycans released from bovine ribonuclease B and the therapeutic monoclonal antibody of trastuzumab.
- Klíčová slova
- Capillary electrophoresis, Fluorescence, Glycan, Labeling, Mass spectrometry, Rhodamine B hydrazide,
- MeSH
- elektroforéza kapilární * metody MeSH
- fluorescenční barviva * chemie MeSH
- fluorescenční spektrometrie metody MeSH
- hmotnostní spektrometrie metody MeSH
- lasery MeSH
- oligosacharidy * chemie analýza MeSH
- polysacharidy * analýza chemie MeSH
- rhodaminy * chemie MeSH
- skot MeSH
- trastuzumab chemie analýza MeSH
- zvířata MeSH
- Check Tag
- skot MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- fluorescenční barviva * MeSH
- oligosacharidy * MeSH
- polysacharidy * MeSH
- rhodamine B MeSH Prohlížeč
- rhodaminy * MeSH
- trastuzumab MeSH
Polymersomes are synthetic vesicles with potential use in healthcare, chemical transformations in confined environment (nanofactories), and in the construction of artificial cells and organelles. In this framework, one of the most important features of such supramolecular structures is the permeability behavior allowing for selective control of mass exchange between the inner and outer compartments. The use of biological and synthetic nanopores in this regard is the most common strategy to impart permeability nevertheless, this typically requires fairly complex strategies to enable porosity. Yet, investigations concerning the permeability of polymer vesicles to different analytes still requires further exploration and, taking these considerations into account, we have detailed investigated the permeability behavior of a variety of polymersomes with regard to different analytes (water, protons, and rhodamine B) which were selected as models for solvents, ions, and small molecules. Polymersomes based on hydrophilic blocks of poly[N-(2-hydroxypropyl)methacrylamide] (PHPMA) or PEO (poly(ethylene oxide)) linked to the non-responsive blocks poly[N-(4-isopropylphenylacetamide)ethyl methacrylate] (PPPhA) or poly(methyl methacrylate) (PMMA), or to the stimuli pH-responsive block poly[2-(diisopropylamino)ethyl methacrylate] (PDPA) have been investigated. Interestingly, the produced PEO-based vesicles are notably larger than the ones produced using PHPMA-containing block copolymers. The experimental results reveal that all the vesicles are inherently permeable to some extent with permeability behavior following exponential profiles. Nevertheless, polymersomes based on PMMA as the hydrophobic component were demonstrated to be the least permeable to the small molecule rhodamine B as well as to water. The synthetic vesicles based on the pH-responsive PDPA block exhibited restrictive and notably slow proton permeability as attributed to partial chain protonation upon acidification of the medium. The dye permeability was evidenced to be much slower than ion or solvent diffusion, and in the case of pH-responsive assemblies, it was demonstrated to also depend on the ionic strength of the environment. These findings are understood to be highly relevant towards polymer selection for the production of synthetic vesicles with selective and time-dependent permeability, and it may thus contribute in advancing biomimicry and nanomedicine.
- Klíčová slova
- Block copolymers, Permeability, Polymersomes, Self-assembly,
- MeSH
- hydrofobní a hydrofilní interakce MeSH
- koncentrace vodíkových iontů MeSH
- permeabilita * MeSH
- polymery * chemie MeSH
- povrchové vlastnosti MeSH
- rhodaminy * chemie MeSH
- umělé buňky chemie MeSH
- velikost částic MeSH
- voda chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- polymery * MeSH
- rhodamine B MeSH Prohlížeč
- rhodaminy * MeSH
- voda MeSH
An in-depth investigation was conducted on a promising composite material (BiVO4/TiO2), focusing on its potential toxicity, photoinduced catalytic properties, as well as its antibiofilm and antimicrobial functionalities. The preparation process involved the synthesis of 2D TiO2 using the lyophilization method, which was subsequently functionalized with sphere-like BiVO4 through wet impregnation. Finally, we developed BiVO4/TiO2 S-scheme heterojunctions which can greatly promote the separation of electron-hole pairs to achieve high photocatalytic performance. The evaluation of concentration- and time-dependent viability inhibition was performed on human lung carcinoma epithelial A549 cells. This assessment included the estimation of glutathione levels and mitochondrial dehydrogenase activity. Significantly, the BiVO4/TiO2 composite demonstrated minimal toxicity towards A549 cells. Impressively, the BiVO4/TiO2 composite exhibited notable photocatalytic performance in the degradation of rhodamine B (k=0.135 min-1) and phenol (k=0.016 min-1). In terms of photoinduced antimicrobial performance, the composite effectively inactivated both gram-negative E. coli and gram-positive E. faecalis bacteria upon 60 minutes of UV-A light exposure, resulting in a significant log 6 (log 10 CFU/mL) reduction in bacterial count. In addition, a 49 % reduction of E. faecalis biofilm was observed. These promising results can be attributed to the unique 2D morphology of TiO2 modified by sphere-like BiVO4, leading to an increased generation of (intracellular) hydroxyl radicals, which plays a crucial role in the treatments of both organic pollutants and bacteria. This research has significant potential for various applications, particularly in addressing environmental contamination and microbial infections.
- Klíčová slova
- 2D TiO2, antibiofilm, antimicrobial, cytotoxicity, photocatalysis, sphere-like BiVO4,
- MeSH
- antibakteriální látky * chemie farmakologie MeSH
- biofilmy účinky léků MeSH
- bismut * chemie MeSH
- buňky A549 MeSH
- Enterococcus faecalis účinky léků MeSH
- Escherichia coli * účinky léků MeSH
- fotochemické procesy * MeSH
- fotolýza MeSH
- katalýza MeSH
- lidé MeSH
- nanostruktury chemie MeSH
- rhodaminy chemie MeSH
- titan * chemie MeSH
- vanadáty * chemie farmakologie MeSH
- viabilita buněk účinky léků MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky * MeSH
- bismut * MeSH
- bismuth vanadium tetraoxide MeSH Prohlížeč
- rhodamine B MeSH Prohlížeč
- rhodaminy MeSH
- titan * MeSH
- titanium dioxide MeSH Prohlížeč
- vanadáty * MeSH
Herein, we introduce versatile molecular tools that enable specific delivery and visualization of photoswitchable lipids at cellular membranes, namely at the plasma membrane and internal membranes. These molecules were prepared by tethering ortho-nitrobenzyl-based fluorescent cages with a signaling lipid bearing an azobenzene photoswitch. They permit two sequential photocontrolled reactions, which are uncaging of a lipid analogue and then its repeated activation and deactivation. We used these molecules to activate GPR40 receptor transiently expressed in HeLa cells and demonstrated downstream modulation of intracellular Ca2+ levels.
- MeSH
- azosloučeniny chemie účinky záření MeSH
- fluorescenční barviva chemie účinky záření MeSH
- fluorescenční mikroskopie MeSH
- HeLa buňky MeSH
- konfokální mikroskopie MeSH
- lidé MeSH
- receptory spřažené s G-proteiny metabolismus MeSH
- rhodaminy chemie účinky záření MeSH
- ultrafialové záření MeSH
- vápník metabolismus MeSH
- zelené fluorescenční proteiny metabolismus MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- azosloučeniny MeSH
- FFAR1 protein, human MeSH Prohlížeč
- fluorescenční barviva MeSH
- receptory spřažené s G-proteiny MeSH
- rhodaminy MeSH
- vápník MeSH
- zelené fluorescenční proteiny MeSH
Humic acids are often regarded as substances with a supramolecular structure which plays an important role in Nature. Their addition into hydrogels can affect their behavior and functioning in different applications. This work is focused on the properties of widely-used hydrogel based on agarose after addition of humic acids-the protonated H-form of humic acids and humic acids with methylated carboxylic groups. Hydrogels enriched by humic acids were studied in terms of their viscoelastic and transport properties. Rotational rheometry and methods employing diffusion cells were used in order to describe the influence of humic acids on the properties and behavior of hydrogels. From the point of view of rheology the addition of humic acids mainly affected the loss modulus corresponding to the relaxation of hydrogel connected with its flow. In the case of diffusion experiments, the transport of dyes (methylene blue and rhodamine) and metal ions (copper and nickel) through the hydrogel was affected by interactions between humic acids and the diffusion probes. The time lag in the hydrogel enriched by humic acids was prolonged for copper, methylene blue and rhodamine. In contrast, the presence of humic acids in hydrogel slightly increased the mobility of nickel. The strongest influence of the methylation of humic acids on diffusion was observed for methylene blue.
- Klíčová slova
- diffusion, humic acid, hydrogel, rheology, secondary structure,
- MeSH
- huminové látky * MeSH
- hydrogely chemie MeSH
- měď chemie MeSH
- methylenová modř chemie MeSH
- nikl chemie MeSH
- reologie MeSH
- rhodaminy chemie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- huminové látky * MeSH
- hydrogely MeSH
- měď MeSH
- methylenová modř MeSH
- nikl MeSH
- rhodaminy MeSH
Nanoparticles have become popular in life sciences in the last few years. They have been produced in many variants and have recently been used in both biological experiments and in clinical applications. Due to concerns over nanomaterial risks, there has been a dramatic increase in investigations focused on safety research. The aim of this paper is to present the advanced testing of rhodamine-derived superparamagnetic maghemite nanoparticles (SAMN-R), which are used for their nontoxicity, biocompatibility, biodegradability, and magnetic properties. Recent results were expanded upon from the basic cytotoxic tests to evaluate cell proliferation and migration potential. Two cell types were used for the cell proliferation and tracking study: mouse embryonic fibroblast cells (3T3) and human mesenchymal stem cells (hMSCs). Advanced microscopic methods allowed for the precise quantification of the function of both cell types. This study has demonstrated that a dose of nanoparticles lower than 20 µg·cm-2 per area of the dish does not negatively affect the cells' morphology, migration, cytoskeletal function, proliferation, potential for wound healing, and single-cell migration in comparison to standard CellTracker™ Green CMFDA (5-chloromethylfluorescein diacetate). A higher dose of nanoparticles could be a potential risk for cytoskeletal folding and detachment of the cells from the solid extracellular matrix.
- Klíčová slova
- cytotoxicity, fibroblast cells, magnetic nanoparticles, mesenchymal stem cells, single-cell migration, wound healing assay,
- MeSH
- biologické markery MeSH
- buněčné linie MeSH
- fibroblasty účinky léků metabolismus MeSH
- imunofenotypizace MeSH
- lidé MeSH
- magnetické nanočástice * chemie MeSH
- mezenchymální kmenové buňky účinky léků metabolismus MeSH
- myši MeSH
- pohyb buněk účinky léků MeSH
- proliferace buněk účinky léků MeSH
- průtoková cytometrie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- rhodaminy chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické markery MeSH
- magnetické nanočástice * MeSH
- reaktivní formy kyslíku MeSH
- rhodaminy MeSH
Here, we describe a fluorescent assay developed to study competitive binding of the glycopeptide antibiotics to live bacteria cells. This assay demonstrated that the mechanism of action of the lipoglycopeptide antibiotics strongly depends on the hydrophobicity of the substitutes, with the best antibacterial activity of the glycopeptide antibiotics equally sharing properties of binding to D-Ala-D-Ala residues of the nascent peptidoglycan and to the membrane.
- MeSH
- antibakteriální látky metabolismus MeSH
- barvení a značení MeSH
- buněčná stěna mikrobiologie MeSH
- Enterococcus faecium metabolismus MeSH
- enterokoky rezistentní vůči vankomycinu metabolismus MeSH
- fluorescence MeSH
- glykopeptidy metabolismus MeSH
- lipoglykopeptidy chemie metabolismus MeSH
- mikrobiální testy citlivosti MeSH
- peptidoglykan metabolismus MeSH
- rhodaminy chemie MeSH
- Staphylococcus aureus metabolismus MeSH
- teikoplanin analogy a deriváty chemie metabolismus MeSH
- vankomycin chemie metabolismus MeSH
- vazba proteinů fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antibakteriální látky MeSH
- dalbavancin MeSH Prohlížeč
- glykopeptidy MeSH
- lipoglykopeptidy MeSH
- oritavancin MeSH Prohlížeč
- peptidoglykan MeSH
- rhodamine isothiocyanate MeSH Prohlížeč
- rhodaminy MeSH
- teikoplanin MeSH
- vankomycin MeSH
A synthetic three-fluorophore system with two enzymatically cleavable linkers has been developed for the simultaneous detection of two proteases in a mixture. The probe was designed to afford single excitation/triple emission ratiometric detection through a fluorescence change during the cleavage of a peptide linker. The developed assays were verified for trypsin and chymotrypsin as the model enzymes.
- MeSH
- aminokumariny chemická syntéza chemie účinky záření MeSH
- chymotrypsin analýza MeSH
- enzymatické testy MeSH
- fenylalanin analogy a deriváty chemie MeSH
- fluoresceiny chemická syntéza chemie účinky záření MeSH
- fluorescence MeSH
- fluorescenční barviva chemická syntéza chemie účinky záření MeSH
- hydrolýza MeSH
- lysin analogy a deriváty chemie MeSH
- rezonanční přenos fluorescenční energie MeSH
- rhodaminy chemická syntéza chemie účinky záření MeSH
- stabilita léku MeSH
- trypsin analýza MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- aminokumariny MeSH
- chymotrypsin MeSH
- fenylalanin MeSH
- fluoresceiny MeSH
- fluorescenční barviva MeSH
- lysin MeSH
- rhodaminy MeSH
- trypsin MeSH
Isoprenoid cytokinins play a number of crucial roles in the regulation of plant growth and development. To study cytokinin receptor properties in plants, we designed and prepared fluorescent derivatives of 6-[(3-methylbut-2-en-1-yl)amino]purine (N6-isopentenyladenine, iP) with several fluorescent labels attached to the C2 or N9 atom of the purine moiety via a 2- or 6-carbon linker. The fluorescent labels included dansyl (DS), fluorescein (FC), 7-nitrobenzofurazan (NBD), rhodamine B (RhoB), coumarin (Cou), 7-(diethylamino)coumarin (DEAC) and cyanine 5 dye (Cy5). All prepared compounds were screened for affinity for the Arabidopsis thaliana cytokinin receptor (CRE1/AHK4). Although the attachment of the fluorescent labels to iP via the linkers mostly disrupted binding to the receptor, several fluorescent derivatives interacted well. For this reason, three derivatives, two rhodamine B and one 4-chloro-7-nitrobenzofurazan labeled iP were tested for their interaction with CRE1/AHK4 and Zea mays cytokinin receptors in detail. We further showed that the three derivatives were able to activate transcription of cytokinin response regulator ARR5 in Arabidopsis seedlings. The activity of fluorescently labeled cytokinins was compared with corresponding 6-dimethylaminopurine fluorescently labeled negative controls. Selected rhodamine B C2-labeled compounds 17, 18 and 4-chloro-7-nitrobenzofurazan N9-labeled compound 28 and their respective negative controls (19, 20 and 29, respectively) were used for in planta staining experiments in Arabidopsis thaliana cell suspension culture using live cell confocal microscopy.
- Klíčová slova
- 6-[(3-methylbut-2-en-1-yl)amino]purine, ARR5:GUS, Competitive receptor bioassay, Cytokinin, Fluorescent label, Fluorescent probe, Isoprenoid, Linker, Live cell confocal microscopy, N(6)-isopentenyladenine,
- MeSH
- 4-chlor-7-nitrobenzofurazan farmakologie MeSH
- adenin analogy a deriváty chemie MeSH
- Arabidopsis metabolismus MeSH
- barvicí látky chemie MeSH
- cytokininy chemie farmakologie MeSH
- fluorescenční barviva chemie MeSH
- isopentenyladenosin chemická syntéza chemie farmakologie MeSH
- karbocyaniny chemie MeSH
- konfokální mikroskopie MeSH
- kukuřice setá metabolismus MeSH
- molekulární struktura MeSH
- proteiny huseníčku metabolismus MeSH
- puriny chemie MeSH
- receptory cytokinové antagonisté a inhibitory chemie MeSH
- regulace genové exprese u rostlin MeSH
- regulátory růstu rostlin metabolismus MeSH
- rhodaminy chemie MeSH
- semenáček metabolismus MeSH
- terpeny metabolismus MeSH
- vývoj rostlin MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 4-chlor-7-nitrobenzofurazan MeSH
- 6-((3-methylbut-2-en-1-yl)amino)purine MeSH Prohlížeč
- adenin MeSH
- barvicí látky MeSH
- cyanine dye 5 MeSH Prohlížeč
- cytokininy MeSH
- fluorescenční barviva MeSH
- isopentenyladenosin MeSH
- karbocyaniny MeSH
- N(6)-(delta(2)-isopentenyl)adenine MeSH Prohlížeč
- N(6),N(6)-dimethyladenine MeSH Prohlížeč
- proteiny huseníčku MeSH
- purine MeSH Prohlížeč
- puriny MeSH
- receptory cytokinové MeSH
- regulátory růstu rostlin MeSH
- rhodamine B MeSH Prohlížeč
- rhodaminy MeSH
- terpeny MeSH
In the present work, we developed a novel needleless emulsion electrospinning technique that improves the production rate of the core/shell production process. The nanofibres are based on poly-ε-caprolactone (PCL) as a continuous phase combined with a droplet phase based on Pluronic F-68 (PF-68). The PCL-PF-68 nanofibres show a time-regulated release of active molecules. Needleless emulsion electrospinning was used to encapsulate a diverse set of compounds to the core phase [i.e. 5-(4,6-dichlorotriazinyl) aminofluorescein -PF-68, horseradish peroxidase, Tetramethylrhodamine-dextran, insulin growth factor-I, transforming growth factor-β and basic fibroblast growth factor]. In addition, the PF-68 facilitates the preservation of the bioactivity of delivered proteins. The system's potential was highlighted by an improvement in the metabolic activity and proliferation of mesenchymal stem cells. The developed system has the potential to deliver susceptible molecules in tissue-engineering applications.
- Klíčová slova
- Pluronic F-68, emulsion electrospinning, growth factor, needleless core/shell electrospinning, time-regulated release,
- MeSH
- biokompatibilní materiály farmakologie MeSH
- dextrany chemie MeSH
- emulze chemie MeSH
- jehly MeSH
- kolagen typ II metabolismus MeSH
- křenová peroxidasa metabolismus MeSH
- mezenchymální kmenové buňky cytologie účinky léků metabolismus MeSH
- mezibuněčné signální peptidy a proteiny farmakologie MeSH
- miniaturní prasata MeSH
- nanovlákna chemie ultrastruktura MeSH
- poloxamer chemie MeSH
- polyestery chemie MeSH
- prasata MeSH
- proteiny aplikace a dávkování MeSH
- rhodaminy chemie MeSH
- tkáňové inženýrství metody MeSH
- tkáňové podpůrné struktury chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- biokompatibilní materiály MeSH
- dextrany MeSH
- emulze MeSH
- kolagen typ II MeSH
- křenová peroxidasa MeSH
- mezibuněčné signální peptidy a proteiny MeSH
- poloxamer MeSH
- polycaprolactone MeSH Prohlížeč
- polyestery MeSH
- proteiny MeSH
- rhodaminy MeSH
- tetramethylrhodamine isothiocyanate MeSH Prohlížeč
