RESUMO
S-Nitroso-N-acetylpenicillamine (SNAP) is among the most common nitric oxide (NO)-donor molecules and its solid-state photolytic decomposition has potential for inhaled nitric oxide (iNO) therapy. The photochemical NO release kinetics and mechanism were investigated by exposing solid-state SNAP to a narrow-band LED as a function of nominal wavelength and intensity of incident light. The photolytic efficiency, decomposition products, and the photolytic pathways of the SNAP were examined. The maximum light penetration depth through the solid layer of SNAP was determined by an optical microscope and found to be within 100-200 µm, depending on the wavelength of light. The photolysis of solid-state SNAP to generate NO along with the stable thiyl (RS·) radical was confirmed using Electron Spin Resonance (ESR) spectroscopy. The fate of the RS· radical in the solid phase was studied both in the presence and absence of O2 using NMR, IR, ESR, and UPLC-MS. The changes in the morphology of SNAP due to its photolysis were examined using PXRD and SEM. The stable thiyl radical formed from the photolysis of solid SNAP was found to be reactive with another adjacent thiyl radical to form a disulfide (RSSR) or with oxygen to form various sulfonyl and sulfonyl peroxyl radicals {RS(O)xO·, x = 0 to 7}. However, the thiyl radical did not recombine with NO to reform the SNAP. From the PXRD data, it was found that the SNAP loses its crystallinity by generating the NO after photolysis. The initial release of NO during photolysis was increased with increased intensity of light, whereas the maximum light penetration depth was unaffected by light intensity. The knowledge gained about the photochemical reactions of SNAP may provide important insight in designing portable photoinduced NO-releasing devices for iNO therapy.
Assuntos
Óxido Nítrico , Espectrometria de Massas em Tandem , S-Nitroso-N-Acetilpenicilamina/farmacologia , Óxido Nítrico/metabolismo , Fotólise , Cromatografia Líquida , Doadores de Óxido Nítrico/química , OxigênioRESUMO
We have designed and synthesized a multifunctional dendritic molecular probe that selectively detects Cu2+ ions via potentiometric and fluorometric techniques with low detection limits (3.5 µM in potentiometry, 15 nM in fluorometry). The selective and reversible binding of the molecule with the Cu2+ ion was used to make a solid-state microsensor (diameter of 25 µm) by incorporating the molecular probe into the carbon-based membrane as an ionophore for Cu(II). The Cu(II) microelectrode has a broad linear range of 10 µM to 1 mM with a near Nernstian slope of 30 mV/log [aCu2+] and detection limit of 3.5 µM. The Cu(II) microsensor has a fast response time (1.5 s), and it has a broad working pH range from 3.5 to 6.0. The incorporation of the hydrophobic dendritic moiety makes the ionophore less prone to leaching in an aqueous matrix for potentiometric measurement. The cinnamaldehyde component of the molecule helps detection of Cu2+ ions fluorometrically, as indicated by a change in fluorescence upon selective and reversible binding of the molecular probe to the Cu2+ ions. The strategic design of the molecular probe allows us to detect Cu2+ ions in drinking water by using this novel dendritic fluoroionophore and solid-state Cu2+ - ion-selective microelectrode.
Assuntos
Água Potável , Sondas Moleculares , Fluorometria , Íons , PotenciometriaRESUMO
We have developed a carbon-based, fast-response potentiometric pH microsensor for use as a scanning electrochemical microscopy (SECM) chemical probe to quantitatively map the microbial metabolic exchange between two bacterial species, commensal Streptococcus gordonii and pathogenic Streptococcus mutans. The 25 µm diameter H+ ion-selective microelectrode or pH microprobe showed a Nernstian slope of 59 mV/pH and high selectivity against major ions such Na+, K+, Ca2+, and Mg2+. In addition, the unique conductive membrane composition aided us in performing an amperometric approach curve to position the probe and obtain a high-resolution pH map of the microenvironment produced by the lactate-producing S. mutans biofilm. The x-directional pH scan over S. mutans also showed the influence of the pH profile on the metabolic activity of another species, H2O2-producing S. gordonii. When these bacterial species were placed in close spatial proximity, we observed an initial increase in the local H2O2 concentration of approximately 12 ± 5 µM above S. gordonii, followed by a gradual decrease in H2O2 concentration (>30 min) to almost zero as lactate was produced, and a subsequent decrease in pH with a more pronounced metabolic output of S. mutans. These results were supported by gene expression and confocal fluorescence microscopic studies. Our findings illustrate that H2O2-producing S. gordonii is dominant while the buffering capacity of saliva is valid (â¼pH 6.0) but is gradually taken over by S. mutans as the latter species slowly starts decreasing the local pH to 5.0 or less by producing lactic acid. Our observations demonstrate the unique capability of our SECM chemical probes for studying real-time metabolic interactions between two bacterial species, which would not otherwise be achievable in traditional assays.
Assuntos
Carbono/química , Peróxido de Hidrogênio/metabolismo , Microscopia Eletroquímica de Varredura/métodos , Streptococcus gordonii/metabolismo , Streptococcus mutans/metabolismo , Alginatos/química , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Técnicas Eletroquímicas , Peróxido de Hidrogênio/análise , Concentração de Íons de Hidrogênio , Ácido Láctico/metabolismo , Microeletrodos , Potássio/química , Sódio/químicaRESUMO
This study presents the utilization of a novel, highly lipophilic nitric oxide (NO) donor molecule, S-nitroso-1-adamantanethiol (SNAT), for developing an NO-emitting polymer surface aimed at preventing thrombus formation and bacterial infection in extracorporeal circuits (ECCs). S-nitroso-1-adamantanethiol, a tertiary nitrosothiol-bearing adamantane species, was synthesized, characterized, and used to impregnate polyvinyl chloride (PVC) tubing for subsequent in vivo evaluation. The impregnation process with SNAT preserved the original mechanical strength of the PVC. In vitro assessments revealed sustained NO release from the SNAT-impregnated PVC tubing (iSNAT), surpassing or matching endothelial NO release levels for up to 42 days. The initial NO release remained stable even after 1 year of storage at -20°C. The compatibility of iSNAT with various sterilization techniques (OPA Plus, hydrogen peroxide, EtO) was tested. Acute in vivo experiments in a rabbit model demonstrated significantly reduced thrombus formation in iSNAT ECCs compared with controls, indicating the feasibility of iSNAT to mitigate coagulation system activation and potentially eliminate the need for systemic anticoagulation. Moreover, iSNAT showed substantial inhibition of microbial biofilm formation, highlighting its dual functionality. These findings underscore the promising utility of iSNAT for long-term ECC applications, offering a multifaceted approach to enhancing biocompatibility and minimizing complications.
RESUMO
A glucose-modified dendritic hydrogel is used as a bioink for bacterial encapsulation. This biocompatible hydrogel is a potentially suitable alternative to conventional alginate hydrogel for bacterial encapsulation, as it readily forms gel in the presence of Na+ or K+ ions without any additional stimuli such as pH, temperature, sonication, or the presence of divalent metal ions. We created a bacterial microhabitat by adding the gelator to phosphate-buffered saline containing live bacteria at physiological pH and using an additive three-dimensional (3D) printing technique. The bacteria remained viable and metabolically active within the 3D printed bacterial microhabitat, as shown with confocal laser scanning microscopy (CLSM) and scanning electrochemical microscopy (SECM).
RESUMO
We designed poly(aryl ether) dendron based transparent hydrogels containing glucose moiety, which undergoes in situ transition from nanofibers to spherical aggregates, upon pH variation. The process is reversible and the assembled structures have been characterized by DLS and SEM. More importantly, efficient dispersion of graphene oxide results in lower CGC (0.08 wt%) value and higher mechanical strength, compared to the native gel.