Styrene-maleic acid (SMA) and similar amphiphilic copolymers are known to cut biological membranes into lipid nanoparticles/nanodiscs containing membrane proteins apparently in their relatively native membrane lipid environment. Our previous work demonstrated that membrane raft microdomains resist such disintegration by SMA. The use of SMA in studying membrane proteins is limited by its heterogeneity and the inability to prepare defined derivatives. In the present paper, we demonstrate that some amphiphilic peptides structurally mimicking SMA also similarly disintegrate cell membranes. In contrast to the previously used copolymers, the simple peptides are structurally homogeneous. We found that their membrane-disintegrating activity increases with their length (reaching optimum at 24 amino acids) and requires a basic primary structure, that is, (XXD)n, where X represents a hydrophobic amino acid (optimally phenylalanine), D aspartic acid, and n is the number of repeats of these triplets. These peptides may provide opportunities for various well-defined potentially useful modifications in the study of membrane protein biochemistry. Our present results confirm a specific character of membrane raft microdomains.
- MeSH
- buněčná membrána metabolismus chemie MeSH
- buněčné linie MeSH
- lidé MeSH
- maleáty chemie MeSH
- membránové mikrodomény metabolismus chemie MeSH
- membránové proteiny * chemie metabolismus MeSH
- peptidy * chemie MeSH
- polystyreny chemie MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
An advantageous alternative to the use of detergents in biochemical studies on membrane proteins are the recently developed styrene-maleic acid (SMA) amphipathic copolymers. In our recent study [1] we demonstrated that using this approach, most T cell membrane proteins were fully solubilized (presumably in small nanodiscs), while two types of raft proteins, GPI-anchored proteins and Src family kinases, were mostly present in much larger (>250 nm) membrane fragments markedly enriched in typical raft lipids, cholesterol and lipids containing saturated fatty acid residues. In the present study we demonstrate that disintegration of membranes of several other cell types by means of SMA copolymer follows a similar pattern and we provide a detailed proteomic and lipidomic characterization of these SMA-resistant membrane fragments (SRMs).
Hydrophobic fibrous slippery liquid-infused porous surfaces (SLIPS) were fabricated by electrospinning polydimethylsiloxane (PDMS) and polystyrene (PS) as a carrier polymer on plasma-treated polyethylene (PE) and polyurethane (PU) substrates. Subsequent infusion of blackseed oil (BSO) into the porous structures was applied for the preparation of the SLIPS. SLIPS with infused lubricants can act as a repellency layer and play an important role in the prevention of biofilm formation. The effect of polymer solutions used in the electrospinning process was investigated to obtain well-defined hydrophobic fibrous structures. The surface properties were analyzed through various optical, macroscopic and spectroscopic techniques. A comprehensive investigation of the surface chemistry, surface morphology/topography, and mechanical properties was carried out on selected samples at optimized conditions. The electrospun fibers prepared using a mixture of PDMS/PS in the ratio of 1:1:10 (g/g/mL) using tetrahydrofuran (THF) solvent showed the best results in terms of fiber uniformity. The subsequent infusion of BSO into the fabricated PDMS/PS fiber mats exhibited slippery behavior regarding water droplets. Moreover, prepared SLIPS exhibited antibacterial activity against Staphylococcus aureus and Escherichia coli bacterium strains.
- MeSH
- dimethylpolysiloxany * MeSH
- Escherichia coli MeSH
- polymery chemie MeSH
- polystyreny * MeSH
- poréznost MeSH
- Publikační typ
- časopisecké články MeSH
In this work, a novel cation exchange membrane, PSEBS SU22 was deployed in microbial fuel cells (MFCs) to examine system efficacy in line with membrane characteristics and inoculum source. It turned out that compared to a reference membrane (Nafion), employing PSEBS SU22 resulted in higher current density and electricity generation kinetics, while the electron recoveries were similar (19-28%). These outcomes indicated more beneficial ion transfer features and lower mass transfer-related losses in the PSEBS SU22-MFCs, supported by membrane water uptake, ion exchange capacity, ionic conductivity and permselectivity. By re-activating the membranes after (bio)foulant removal, PSEBS SU22 regained nearly its initial conductivity, highlighting a salient functional stability. Although the particular inoculum showed a clear effect on the microbial composition of the membrane biofouling layers, the dominance of aerobic species was revealed in all cases. Considering all the findings, the PSEBS SU22 seems to be promising for application in MFCs.
Photodynamic inactivation (PDI) is a promising approach for the efficient killing of pathogenic microbes. In this study, the photodynamic effect of sulfonated polystyrene nanoparticles with encapsulated hydrophobic 5,10,15,20-tetraphenylporphyrin (TPP-NP) photosensitizers on Gram-positive (including multi-resistant) and Gram-negative bacterial strains was investigated. The cell viability was determined by the colony forming unit method. The results showed no dark cytotoxicity but high phototoxicity within the tested conditions. Gram-positive bacteria were more sensitive to TPP-NPs than Gram-negative bacteria. Atomic force microscopy was used to detect changes in the morphological properties of bacteria before and after the PDI treatment.
- MeSH
- Bacteria účinky léků účinky záření MeSH
- fotochemické procesy * MeSH
- fotochemoterapie metody MeSH
- mikroskopie atomárních sil MeSH
- nanočástice * chemie MeSH
- polystyreny * chemie MeSH
- porfyriny aplikace a dávkování chemie MeSH
- příprava léků * MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
The aim of our research was to study the behaviour of adipose tissue-derived stem cells (ADSCs) and vascular smooth muscle cells (VSMCs) on variously modified poly(L-lactide) (PLLA) foils, namely on pristine PLLA, plasma-treated PLLA, PLLA grafted with polyethylene glycol (PEG), PLLA grafted with dextran (Dex), and the tissue culture polystyrene (PS) control. On these materials, the ADSCs were biochemically differentiated towards VSMCs by a medium supplemented with TGFβ1, BMP4 and ascorbic acid (i.e. differentiation medium). ADSCs cultured in a non-differentiation medium were used as a negative control. Mature VSMCs cultured in both types of medium were used as a positive control. The impact of the variously modified PLLA foils and/or differences in the composition of the medium were studied with reference to cell adhesion, growth and differentiation. We observed similar adhesion and growth of ADSCs on all PLLA samples when they were cultured in the non-differentiation medium. The differentiation medium supported the expression of specific early, mid-term and/or late markers of differentiation (i.e. type I collagen, αSMA, calponin, smoothelin, and smooth muscle myosin heavy chain) in ADSCs on all tested samples. Moreover, ADSCs cultured in the differentiation medium revealed significant differences in cell growth among the samples that were similar to the differences observed in the cultures of VSMCs. The round morphology of the VSMCs indicated worse adhesion to pristine PLLA, and this sample was also characterized by the lowest cell proliferation. Culturing VSMCs in the differentiation medium inhibited their metabolic activity and reduced the cell numbers. Both cell types formed the most stable monolayer on plasma-treated PLLA and on the PS control. The behaviour of ADSCs and VSMCs on the tested PLLA foils differed according to the specific cell type and culture conditions. The suitable biocompatibility of both cell types on the tested PLLA foils seems to be favourable for vascular tissue engineering purposes.
- MeSH
- aorta metabolismus MeSH
- biokompatibilní materiály MeSH
- biopolymery chemie MeSH
- buněčná adheze MeSH
- buněčná diferenciace účinky léků MeSH
- kmenové buňky cytologie MeSH
- mikroskopie atomárních sil MeSH
- myocyty hladké svaloviny cytologie MeSH
- oxaziny chemie MeSH
- polyestery chemie MeSH
- polymery chemie MeSH
- polysacharidy chemie MeSH
- polystyreny chemie MeSH
- povrchové vlastnosti MeSH
- prasata MeSH
- proliferace buněk MeSH
- svaly hladké cévní cytologie MeSH
- testování materiálů MeSH
- tkáňové inženýrství metody MeSH
- tuková tkáň metabolismus MeSH
- xantheny chemie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Currently, widely available three-dimensional (3D) printers are very popular with the public. Previous research has shown that these printers can emit ultrafine particles (UFPs) and volatile organic compounds (VOCs). Several studies have examined the emissivity of filaments from 3D printing, except glycol modified polyethylene terephthalate (PETG) and styrene free co-polyester (NGEN) filaments. The aim of this study was to evaluate UFP and VOC emissions when printing using a commonly available 3D printer (ORIGINAL PRUSA i3 MK2 printer) using PETG and NGEN. The concentrations of UFPs were determined via measurements of particle number concentration and size distribution. A thermal analysis was carried out to ascertain whether signs of fiber decomposition would occur at printing temperatures. The total amount of VOCs was determined using a photoionization detector, and qualitatively analyzed via gas chromatography-mass spectrometry. The total particle concentrations were 3.88 × 1010 particles for PETG and 6.01 × 109 particles for NGEN. VOCs at very low concentrations were detected in both filaments, namely ethylbenzene, toluene, and xylene. In addition, styrene was identified in PETG. On the basis of our results, we recommend conducting additional measurements, to more accurately quantify personal exposure to both UFPs and VOCs, focusing on longer exposure as it can be a source of potential cancer risk.
This study presents a proof of principle concept for a two-dimensional bioprinted glucose sensor on Petri dishes that allows for glucose measurements in cell culture medium. To improve bioink adhesion, the polystyrene surfaces of standard Petri dishes are activated with argon plasma, which increases roughness and hydrophilicity. The bioink containing the sensor chemistry - namely fluorescently labeled ConA/Dextran embedded in alginate microbeads - was printed on the activated Petri dishes with an extrusion-based bioprinter. The printed sensor showed good stability and adhesive properties on polystyrene. The glucose concentration was examined using a standard fluorescence microscope with filters adapted to the emission wavelength of the donor and reference dyes. The printed glucose sensor showed high sensitivity and good linearity in a physiologically relevant range of glucose concentrations.
An emerging alternative to the use of detergents in biochemical studies on membrane proteins is apparently the use styrene-maleic acid (SMA) amphipathic copolymers. These cut the membrane into nanodiscs (SMA-lipid particles, SMALPs), which contain membrane proteins possibly surrounded by their native lipid environment. We examined this approach for studies on several types of T cell membrane proteins, previously defined as raft or non-raft associated, to see whether the properties of the raft derived SMALPs differ from non-raft SMALPs. Our results indicate that two types of raft proteins, GPI-anchored proteins and two Src family kinases, are markedly present in membrane fragments much larger (>250 nm) than those containing non-raft proteins (<20 nm). Lipid probes sensitive to membrane fluidity (membrane order) indicate that the lipid environment in the large SMALPs is less fluid (more ordered) than in the small ones which may indicate the presence of a more ordered lipid Lo phase which is characteristic of membrane rafts. Also the lipid composition of the small vs. large SMALPs is markedly different - the large ones are enriched in cholesterol and lipids containing saturated fatty acids. In addition, we confirm that T cell membrane proteins present in SMALPs can be readily immunoisolated. Our results support the use of SMA as a potentially better (less artifact prone) alternative to detergents for studies on membrane proteins and their complexes, including membrane rafts.
- MeSH
- anizotropie MeSH
- buněčná membrána chemie MeSH
- cholesterol chemie MeSH
- detergenty chemie MeSH
- gelová chromatografie MeSH
- Jurkat buňky MeSH
- lidé MeSH
- lipidové dvojvrstvy chemie MeSH
- lipidy chemie MeSH
- maleáty chemie MeSH
- mastné kyseliny chemie MeSH
- membránové mikrodomény chemie MeSH
- membránové proteiny chemie MeSH
- membrány umělé MeSH
- myši inbrední C57BL MeSH
- myši MeSH
- polymery chemie MeSH
- radiační rozptyl MeSH
- rozpustnost MeSH
- styren chemie MeSH
- světlo MeSH
- T-lymfocyty cytologie MeSH
- ultracentrifugace MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Bark beetles kill apparently vigorous conifers during epidemics by means of pheromone-mediated aggregation. During non-endemic conditions the beetles are limited to use trees with poor defense, like wind-thrown. To find olfactory cues that help beetles to distinguish between trees with strong or weak defense, we collected volatiles from the bark surface of healthy felled or standing Picea abies trees. Furthermore, living trees were treated with methyl jasmonate in order to induce defense responses. Volatiles were analyzed by combined gas chromatography and electroantennographic detection (GC-EAD) on Ips typographus antennae. Compounds eliciting antennal responses were characterized by single sensillum recording for identification of specific olfactory sensory neurons (OSN). Release of monoterpene hydrocarbons decreased, while oxygenated compounds increased, from spring to early summer in felled trees. In both beetle sexes particular strong EAD activity was elicited by trace amounts of terpene alcohols and ketones. 4-Thujanol gave a very strong response and the absolute configuration of the tested natural product was assigned to be (+)-trans-(1R,4S,5S)-thujanol by stereoselective synthesis and enantioselective gas chromatography. One type of OSN responded to all ketones and five other OSN were characterized by the type of compounds that elicited responses. Three new OSN classes were found. Of the eight EAD-active compounds found in methyl jasmonate-treated bark, the known anti-attractant 1,8-cineole was the one most strongly induced. Our data support the hypothesis that highly active oxygenated host volatiles could serve as positive or negative cues for host selection in I. typographus and in other bark beetles.
- MeSH
- acetáty farmakologie MeSH
- brouci fyziologie MeSH
- cyklopentany farmakologie MeSH
- elektrofyziologické jevy účinky léků MeSH
- kůra rostlin chemie účinky léků metabolismus MeSH
- monoterpeny chemická syntéza chemie farmakologie MeSH
- oxylipiny farmakologie MeSH
- plynová chromatografie s hmotnostně spektrometrickou detekcí MeSH
- smrk chemie metabolismus MeSH
- stereoizomerie MeSH
- styren chemie farmakologie MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
