Conformational Space of the Translocation Domain of Botulinum Toxin: Atomistic Modeling and Mesoscopic Description of the Coiled-Coil Helix Bundle.

The toxicity of botulinum multi-domain neurotoxins (BoNTs) arises from a sequence of molecular events, in which the translocation of the catalytic domain through the membrane of a neurotransmitter vesicle plays a key role. A recent structural study of the translocation domain of BoNTs suggests that the interaction with the membrane is driven by the transition of an α helical switch towards a ß hairpin. Atomistic simulations in conjunction with the mesoscopic Twister model are used to investigate the consequences of this proposition for the toxin-membrane interaction. The conformational mobilities of the domain, as well as the effect of the membrane, implicitly examined by comparing water and water-ethanol solvents, lead to the conclusion that the transition of the switch modifies the internal dynamics and the effect of membrane hydrophobicity on the whole protein. The central two α helices, helix 1 and helix 2, forming two coiled-coil motifs, are analyzed using the Twister model, in which the initial deformation of the membrane by the protein is caused by the presence of local torques arising from asymmetric positions of hydrophobic residues. Different torque distributions are observed depending on the switch conformations and permit an origin for the mechanism opening the membrane to be proposed.

Optimization of Bioactive Compound Extraction from Saffron Petals Using Ultrasound-Assisted Acidified Ethanol Solvent: Adding Value to Food Waste.

The saffron industry produces large by-products, including petals with potential bioactive compounds, which are cheap and abundant, making them an attractive alternative to expensive stigmas for extracting bioactive components. This study aimed to optimize the extraction conditions of bioactive compounds from vacuum-dried saffron petals using an ultrasound-assisted acidified ethanol solvent. Three factors were considered: ethanol concentration (0-96%), citric acid concentration in the final solvent (0-1%), and ultrasound power (0-400 watt). This study examined the effects of these factors on parameters like maximum antioxidant activity, total anthocyanin content, total phenolic content, and the total flavonoid content of the extraction. This study found that saffron petal extract's antioxidant activity increases with higher ethanol concentration, citric acid dose, and ultrasound power, but that an increased water content leads to non-antioxidant compounds. Increasing the dosage of citric acid improved the extraction of cyanidin-3-glucoside at different ultrasound power levels. The highest extraction was achieved with 400 watts of ultrasound power and 1% citric acid. Ethanol concentration did not affect anthocyanin extraction. Higher ethanol concentration and greater citric acid concentration doses resulted in the maximum extraction of total phenolic content, with a noticeable drop in extraction at higher purity levels. This study found that increasing the proportion of citric acid in the final solvent did not affect flavonoid extraction at high ethanol concentration levels, and the highest efficiency was observed at 200 watts of ultrasound power. The optimum values of the independent parameters for extracting bioactive compounds from saffron petals included 96% ethanol concentration, 0.67% citric acid concentration, and 216 watts of ultrasound power, resulting in a desirability value of 0.82. This ultrasound-assisted acidified ethanolic extract can be used in the food industry as a natural antioxidant and pigment source.

Pulp fibroblast cell apoptosis after application of hema dentine bonding material with ethanol and water solvent

Abstract The most common main materials for dentin bonding for composite resin restoration is 2-hydroxyethyl methacrylate (HEMA). HEMA has beneficial physical and chemical properties, and stable, yet toxic. The addition of ethanol or water, may reduce the toxic effect of HEMA. Ethanol solvent has lower H-bonding capacity compared to water solvent, so it can bind less free radicals from the residual monomer. This study aimed to analyze apoptosis due to dentine bonding application with ethanol and water solvent. Fibroblast culture cells were obtained from extracted third molar, by means of tripsinasion method. The cells were divided into 4 groups as reached confluent: cell culture without treatment as control, cell culture with scaffold chitosan, cell culture with scaffold and polymerized dentin bonding with ethanol or water solvent. Apoptosis observation was conducted using immunohistochemistry method with ethidium bromide acridin orange staining, under fluorescent microscope with 40´ magnification. There was a significant difference among groups (p=0.0001), yet no differences found between different solvent. Apoptosis rate in fibroblast cells culture exposed to HEMA bonding with ethanol solvent was 67%, while the cells exposed to HEMA bonding with water solvent was 44%. The effect of dentin bonding with ethanol solvent and water solvent towards apoptosis rate of pulp fibroblast cells is not different.

Resumo Os principais materiais para adesão dentinária em restaurações de resina composta são o 2-hidroxietil metacrilato (HEMA). O HEMA possui propriedades físicas e químicas benéficas e estáveis, ainda que tóxicas. A adição de etanol ou água pode reduzir o efeito tóxico do HEMA. O solvente etanol possui uma menor capacidade de ligação H em comparação com o solvente água, de modo que pode ligar menos radicais livres do monômero residual. Este trabalho teve como objetivo analisar a apoptose pela aplicação de adesivos dentinários com solventes etanol e água. Células de cultura de fibroblastos foram obtidas a partir do terceiro molar extraído, por meio do método de tripsinaion. As células foram divididas em 4 grupos como confluentes: cultura celular sem tratamento como controle, cultura celular com arcabouço de quitosana, cultura celular com arcabouço e adesivo dentinário polimerizado com solvente etanol ou água. A observação da apoptose foi realizada utilizando o método imunohistoquímico com coloração com brometo de etídio e acridina laranja, sob microscópio de fluorescência com aumento de 40´. Houve uma diferença significativa entre os grupos (p = 0,0001), mas não houve diferenças entre os solventes. A taxa de apoptose em cultura de células de fibroblastos expostos à adesão baseada em HEMA com solvente etanol foi de 67%, enquanto as células expostas à adesão baseada em HEMA com solvente de água foi de 44%. O efeito da adesão dentinária com solvente etanol e solvente água sobre a taxa de apoptose de células de fibroblastos de polpa não é diferente.

Stretchable PDMS Embedded Fibrous Membranes Based on an Ethanol Solvent System for Waterproof and Breathable Applications.

Flexible membranes with excellent waterproofness and breathability have been strongly desired in wound dressing applications with the aim of providing both protection and comfort. Despite the advances in protective clothing using waterproof breathable materials, the construction of waterproof breathable membranes suited for wound dressing still faces huge challenges to eliminate the toxic solvent residue-related harm to health and improve the waterproof, breathable, and stretchable performance. In the current work, we developed a facile and versatile approach based on one-step electrospinning and an ethanol solvent system for producing skinlike waterproof and breathable polydimethylsiloxane (PDMS) embedded polyvinyl butyral (PVB) fibrous membranes. Benefiting from the addition of hydrophobic agent PDMS, a reduced maximum pore size and enhanced surface hydrophobicity were achieved, contributing to a maximum hydrostatic pressure of 54.32 kPa, which was about 4.0 times that of the PVB fibrous membrane. In addition, the obtained PVB/PDMS fibrous membranes exhibited a remarkable water vapor transmission rate of 8.98 kg m-2 day-1, and enhanced mechanical strength of 4.95 MPa. The developed fibrous membranes provided human skinlike functions, including blocking liquid water penetration inside, permitting water vapor transmission outside, and allowing for sufficient stretching at joint positions. Taken together, this work could contribute to a better design of health-friendly, waterproof, breathable, and stretchable wound dressing materials.

The insight study of SnO pico size particles in an ethanol-water system followed by its biosensing application.

Pico sized Stannous oxide particles (SnO PPs) were synthesized in an ethanol-water solvent system on the surface of nitrogen doped graphene oxide (GO). The highly conductive support was a combination of dual interactions between 4-aminomethylbenzylamine (AMBA) and GO. The oppositely positioned -NH2 linkers of the AMBA were covalently incorporated into the GO matrix through condensation reaction followed by the strong π - π stacking interactions between aromatic rings of AMBA and GO. The change in the local chemical environment of GO via dual interactions provided a suitable atmosphere for the growth and dispersion of SnO PPs on GO-AMBA surface. The possible mechanism for the formation of SnO in an ethanol-water solvent system was evaluated. Furthermore, a light was shed on the factors responsible for the pico size of SnO particles synthesis along with its phenomenal distribution on the GO-AMBA surface. The catalyst containing SnO PPs was deployed as a biosensor for the detection of ascorbic acid (AA) for the very first time. A very wide linear range of 5.0 × 10-5-7.0 × 10-3 M, limit of detection (LOD) of 1.19 × 10-5 M along with excellent practical feasibility, storage stability, repeatability and selectivity towards AA electrooxidation showed the excellent synergy between nitrogen-rich GO surface and SnO PPs. The sensitivity (885.54 µAmM-1cm-2) of the catalyst was the most attractive feature, as it was obtained in the presence of 5 and 2-fold higher concentration of UA and DA interfering species respectively.