RESUMO
Bioluminescent gold nanoparticles (AuNPs) were synthesized in situ using dithiol-terminated polyethylene glycol (PEG(SH)2) as reducer and stabilizing agents. Hybrid Au/F3O4 nanoparticles were also produced in a variation of synthesis, and both types of nanostructures had the polymer capping replaced by L-cysteine (Cys). The four types of nanoparticles, PEG(SH)2AuNPs, PEG(SH)2Au/F3O4NPs, CysAuNPs, and CysAu/F3O4NPs were associated with purified recombinant Pyrearinus termitilluminans green emitting click beetle luciferase (PyLuc) and Phrixotrix hirtus (RELuc) red-emitting railroad worm luciferase. Enzyme association with PEG(SH)2 was also investigated as a control. Luciferases were chosen because they catalyze bioluminescent reactions used in a wide range of bioanalytical applications, including ATP assays, gene reporting, high-throughput screening, bioluminescence imaging, biosensors and other bioluminescence-based assays. The immobilization of PyLuc and RELuc promoted partial suppression of the enzyme luminescence activity in a functionalization-dependent way. Association of PyLuc and RELuc with AuNPs increased the enzyme operational stability in relation to the free enzyme, as evidenced by the luminescence intensity from 0 to 7 h after substrate addition. The stability of the immobilized enzymes was also functionalization-dependent and the association with CysAuNPs was the condition that combined more sustained luminescent activity with a low degree of luminescence quenching. The higher enzymatic stability and sustained luminescence of luciferases associated with nanoparticles may improve the applicability of bioluminescence for bioimaging and biosensing purposes.
Assuntos
Besouros , Nanopartículas Metálicas , Animais , Ouro , Luciferases/genética , Luminescência , Medições LuminescentesRESUMO
The development of self-disinfectant devices is highly needed to prevent and control infections, mainly caused by virus. In the past years, coronaviruses have been a threat to humanity, causing severe epidemics of respiratory infections such as severe acute respiratory syndrome (SARS), in 2003, and Middle East respiratory syndrome (MERS) in 2012, and presently the SARS-CoV2 is causing the COVID-19 pandemic. Previous studies have demonstrated that surface contamination play a significant role in the spreading of viruses. These studies demonstrated that the production of highly reactive species by copper alloys contributes to rapid elimination of viruses. Nanostructured materials such as semiconductors TiO2, Co3O4 CuO, NiO, and TiO2, and silver nanoparticles can decrease the virus viability on the surfaces when associated with polymers and textiles, especially in conditions of light exposure. In addition, graphene oxide is rising as a promising material for inactivation of viruses due to its capacity of destroying the viral envelope and capsid. The virucidal property of these materials can be enhanced by increasing their functionalization with photosensitizers. The present mini-review brings subsidies for the development of new advanced self-disinfectant materials that can be used in the manufacture of gloves, masks, and a variety of other devices.
Assuntos
Infecções por Coronavirus , Nanopartículas Metálicas , Coronavírus da Síndrome Respiratória do Oriente Médio , Pandemias , Pneumonia Viral , Betacoronavirus , COVID-19 , Humanos , SARS-CoV-2 , PrataRESUMO
In this study, we prepared translucid hydrogels with different concentrations of silk fibroin, extracted from raw silk fibers, and used them as a matrix to incorporate the photosensitizer 5-(4-aminophenyl)-10,15,20-tris-(4-sulphonatophenyl) porphyrin trisodium for application in photodynamic therapy (PDT). The hydrogels obtained were characterized by rheology, spectrophotometry, and scattering techniques to elucidate the factors involved in the formation of the hydrogel, and to characterize the behavior of silk fibroin (SF) after incorporating of the porphyrin to the matrix. The rheology results demonstrated that the SF hydrogels had a shear thinning behavior. In addition, we were able to verify that the structure of the material was able to be recovered over time after shear deformation. The encapsulation of porphyrins in hydrogels leads to the formation of self-assembled peptide nanostructures that prevent porphyrin aggregation, thereby greatly increasing the generation of singlet oxygen. Also, our findings suggest that porphyrin can diffuse out of the hydrogel and permeate the outer skin layers. This evidence suggests that SF hydrogels could be used as porphyrin encapsulation and as a drug carrier for the sustained release of photosensitizers for PDT.
Assuntos
Fibroínas/química , Hidrogéis/química , Dicroísmo Circular , Portadores de Fármacos/química , Luz , Fotoquimioterapia , Fármacos Fotossensibilizantes/química , Fármacos Fotossensibilizantes/metabolismo , Porfirinas/química , Porfirinas/metabolismo , Reologia , Oxigênio Singlete/metabolismoRESUMO
Bromelain (Bro) is a multiprotein complex extracted from the pineapple plant Ananas comosus, composed of at least eight cysteine proteases. Bro has a wide range of applications in medicine and industry, where the stability of its active proteases is always a major concern. The present study describes the improvement of stability and gain of specific activity in the enzymatic content of Bro immobilized on gold nanoparticles (GNPs). GNPs were synthesized in situ using Bro as the reducing and stabilizing agents and characterized by surface plasmon resonance and transmission electron microscopy. Consistent with the structural changes observed by circular dichroism analysis, the association with GNPs affected enzyme activity. The active Bro immobilized on GNPs (NanoBro) remained stable under storage and gained thermal stability consistent with a thermophilic enzyme. Proteolytic assays were performed on type I collagen membranes using fluorescence spectroscopy of O-phthaldialdehyde (OPA), changes in the membrane superficial structure, and topography by scanning electron microscopy, FTIR, and scanning laser confocal microscopy. Another characteristic of the NanoBro observed was the significant increase in susceptibility to the inhibitory effect of E-64, indicating a gain in cysteine protease activity. The higher stability and specific activity of NanoBro contributed to the broadening and improvement of Bro applications.
RESUMO
Manganese porphyrins are well-known protectors against the deleterious effects of pro-oxidant species such as superoxide ions and hydrogen peroxide. The present study investigated the antioxidant cytochrome c-like activities of Mn(III)TMPyP [meso-tetrakis (4-N-methyl pyridinium) porphyrin] against superoxide ion and hydrogen peroxide that remained unexplored for this porphyrin. The association of TMPyP with a model of the inner mitochondrial membrane, cardiolipin (CL)-containing liposomes, shifted +30 mV vs. NHE (normal hydrogen electrode) redox potential of the Mn(II)/Mn(III) redox couple. In CL-containing liposomes, Mn(III)TMPyP was reduced by superoxide ions and recycled by Fe(III)cytochrome c to the oxidized form. Similarly, isolated rat liver mitoplasts added to a sample of Mn(II)TMPyP promoted immediate porphyrin reoxidation by electron transfer to the respiratory chain. These results show that Mn(III)TMPyP can act as an additional pool of Fe(III)cytochrome c capable of transferring electrons that escape from the IV complex back into the respiratory chain. Unlike Fe(II)cytochrome c, Mn(II)TMPyP was not efficient for hydrogen peroxide clearance. Therefore, by reducing cytochrome c, Mn(II)TMPyP can indirectly contribute to hydrogen peroxide elimination.
Assuntos
Antioxidantes/química , Citocromos c/química , Mitocôndrias Hepáticas/enzimologia , Membranas Mitocondriais/enzimologia , Porfirinas/química , Animais , Ratos , Ratos WistarRESUMO
The use of ultraviolet (UV) and blue irradiation to sterilize surfaces is well established, but commercial applications would be enhanced if the light source is replaced with ambient light. In this paper, it is shown that nanofibers can be explored as an alternative methodology to UV and blue irradiation for bacterial inactivation. It is demonstrated that this is indeed possible using spun nanofibers of poly[lactic-co-(glycolic acid)] (PLGA). This work shows that PLGA spun scaffolds can promote photoinactivation of Staphylococcus aureus and Escherichia coli bacteria with ambient light or with laser irradiation at 630 nm. With the optimized scaffold composition of PLGA85:15 nanofibers, the minimum intensity required to kill the bacteria is much lower than in antimicrobial blue light applications. The enhanced effect introduced by PLGA scaffolds is due to their nanofiber structures since PLGA spun nanofibers were able to inactivate both S. aureus and E. coli bacteria, but cast films had no effect. These findings pave the way for an entirely different method to sterilize surfaces, which is less costly and environmentally friendly than current procedures. In addition, the scaffolds could also be used in cancer treatment with fewer side effects since photosensitizers are not required.
Assuntos
Eletricidade , Escherichia coli/fisiologia , Viabilidade Microbiana/efeitos dos fármacos , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/farmacologia , Staphylococcus aureus/fisiologia , Raios Ultravioleta , Escherichia coli/efeitos dos fármacos , Escherichia coli/efeitos da radiação , Viabilidade Microbiana/efeitos da radiação , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/efeitos da radiaçãoRESUMO
Solid-state ionic conductor is an essential and critical part of electrochemical devices such as batteries and sensors. Nano-sized silver iodide (AgI) is the most promising ionic conductor due to its superionic conductivity at room temperature. In recent years, proteins have been used as organic templates to obtain high-performance solid-state ionic conductors as well as to extend their applications in a biosensor. Here, we report the unprecedented ultrafast synthesis of thermally stable protein-coated AgI nanoparticles (NPs) through the photo-irradiation method for solid-state electrolyte. The synthesis was performed using a hyperthermostable bacterial ß-glucosidase. The protein-coated AgI NPs with an approximate diameter of 13 nm showed that the controllable transition from the α- to ß-/γ-phase was drastically suppressed down to 41 °C in the cooling process. After drying, the product represents a thermally stable organic-inorganic hybrid system with superionic conductivity. It is noteworthy that the superionic conductivity (σ Ë 0.14 S/cm at 170 °C) of thermally stable protein-coated AgI NPs is maintained during several thermal cycles (25-170 °C). To our knowledge, this is the first report showing the diffusion of mobile Ag+ ions on the surface of the AgI NPs through a protein matrix. The facile synthesis method and high performance of the protein-coated AgI NPs may provide a latent application in the mass production of nanobatteries and other technological applications.
Assuntos
Iodetos/química , Nanopartículas/química , Compostos de Prata/química , Temperatura , beta-Glucosidase/química , Condutividade Elétrica , Bactérias Anaeróbias Gram-Negativas/enzimologia , Microscopia Eletrônica de Transmissão , Tamanho da Partícula , Propriedades de Superfície , beta-Glucosidase/metabolismoRESUMO
Ca2+-overload contributes to the oxidation of mitochondrial membrane lipids and associated events such as the permeability transition pore (MPTP) opening. Numerous experimental studies about the Ca2+/cardiolipin (CL) interaction are reported in the literature, but there are few studies in conjunction with theoretical approaches based on ab initio calculations. In the present study, the lipid fraction of the inner mitochondrial membrane was modeled as POPC/CL large unilamellar vesicles (LUVs). POPC/CL and, comparatively, POPC, and CL LUVs were challenged by singlet molecular oxygen using the anionic porphyrin TPPS4 as a photosensitizer and by free radicals produced by Fe2+-citrate. Calcium ion favored both types of lipid oxidation in a lipid composition-dependent manner. In membranes containing predominantly or exclusively POPC, Ca2+ increased the oxidation at later reaction times while the oxidation of CL membranes was exacerbated at the early times of reaction. Considering that Ca2+ interaction affects the lipid structure and packing, density functional theory (DFT) calculations were applied to the Ca2+ association with totally and partially protonated and deprotonated CL, in the presence of water. The interaction of totally and partially protonated CL head groups with Ca2+ decreased the intramolecular P-P distance and increased the hydrophobic volume of the acyl chains. Consistently with the theoretically predicted effect of Ca2+ on CL, in the absence of pro-oxidants, giant unilamellar vesicles (GUVs) challenged by Ca2+ formed buds and many internal vesicles. Therefore, Ca2+ induces changes in CL packing and increases the susceptibility of CL to the oxidation promoted by free radicals and excited species.