RESUMO
AIM: The antifungal activity was studied on sessile and persister cells (PCs) of Candida tropicalis biofilms of gold nanoparticles (AuNPs) stabilized with cetyltrimethylammonium bromide (CTAB-AuNPs) and those conjugated with cysteine, in combination with Amphotericin B (AmB). MATERIALS/METHODS: The PC model was used and synergistic activity was tested by the checkerboard assay. Biofilms were studied by crystal violet and scanning electron microscopy. RESULTS/CONCLUSIONS: After the combination of both AuNPs and AmB the biofilm biomass was reduced, with significant differences in architecture being observed with a reduced biofilm matrix. In addition, the CTAB-AuNPs-AmB combination significantly reduced PCs. Understanding how these AuNPs aid in the fight against biofilms and the development of new approaches to eradicate PCs has relevance for chronic infection treatment.
Assuntos
Anfotericina B , Antifúngicos , Biofilmes , Candida tropicalis , Sinergismo Farmacológico , Ouro , Nanopartículas Metálicas , Testes de Sensibilidade Microbiana , Candida tropicalis/efeitos dos fármacos , Ouro/química , Ouro/farmacologia , Biofilmes/efeitos dos fármacos , Anfotericina B/farmacologia , Anfotericina B/química , Nanopartículas Metálicas/química , Antifúngicos/farmacologia , Antifúngicos/química , Cetrimônio/química , Compostos de Cetrimônio/farmacologia , Compostos de Cetrimônio/químicaRESUMO
This study presents the results of the electrochemical degradation of the emulsifier cetrimonium chloride (CTAC) on a boron-doped diamond (BDD) anode under different current densities and flow rates. Higher values of these parameters result in a more rapid removal. Nevertheless, operation at low current reduces the required applied charge and increases the chemical oxygen demand (COD) removal efficiency, as there is less development of ineffective parasitic reactions. On the other hand, high flow rates reduce the required volumetric applied charge and increase the COD removal current efficiency. In order to assist and enrich the study, an economic analysis has been performed. For short expected plant lifespans, operation at low current is advantageous due to the lower investment required, whereas for longer expected lifespans, the operational costs make the lower current densities less costly. High flow rates are always advantageous from a financial point of view.
Assuntos
Compostos de Cetrimônio/química , Diamante , Poluentes Químicos da Água , Boro , Cetrimônio , Eletroquímica , Eletrodos , Oxirredução , Purificação da ÁguaRESUMO
Numerous applications have been described for microperoxidases (MPs) such as in photoreceptors, sensing, drugs, and hydrogen evolution. The last application was obtained by replacing Fe(III), the native central metal, by cobalt ion and inspired part of the present study. Here, the Fe(III) of MP-11 was replaced by Cu(II) that is also a stable redox state in aerated medium, and the structure and activity of both MPs were modulated by the interaction with the positively charged interfaces of lipids. Comparative spectroscopic characterization of Fe(III) and Cu(II)MP-11 in the studied media demonstrated the presence of high and low spin species with axial distortion. The association of the Fe(III)MP-11 with CTAB and Cu(II)MP-11 with DODAB affected the colloidal stability of the surfactants that was recovered by heating. This result is consistent with hydrophobic interactions of MPs with DODAB vesicles and CTAB micelles. The hydrophobic interactions decreased the heme accessibility to substrates and the Fe(III) MP-11catalytic efficiency. Cu(II)MP-11 challenged by peroxides exhibited a cyclic Cu(II)/Cu(I) interconversion mechanism that is suggestive of a mimetic Cu/ZnSOD (superoxide dismutase) activity against peroxides. Hydrogen peroxide-activated Cu(II)MP-11 converted Amplex Red® to dihydroresofurin. This study opens more possibilities for technological applications of MPs.
Assuntos
Cobre/química , Compostos Férricos/química , Peroxidases/química , Catálise , Cetrimônio , Compostos de Cetrimônio/química , Heme/química , Peróxido de Hidrogênio/química , Interações Hidrofóbicas e Hidrofílicas , Lipossomos/química , Micelas , Oxirredução , Ligação Proteica , Conformação Proteica , Compostos de Amônio Quaternário/química , Superóxido Dismutase/química , Propriedades de SuperfícieRESUMO
Electrophoresis under denaturing conditions in the presence of SDS is a standard method for the protein and enzyme scientist. Nevertheless, there are special situations where this method may originate nonoptimal results. SDS may cause protein aggregation or precipitation. Beyond this, depending on the type of protein, some just do not resolve well or migrate abnormally in SDS gels. SDS, an anionic detergent, may be however substituted by a cationic detergent, like CTAB (cetyltrimethylammonium bromide), in order to solubilize and electrophorize proteins. CTAB electrophoresis allows the separation of proteins based on molecular weight and can be carried out at neutral or acidic pH. Here, we describe the development of a CTAB zymography method to analyze aspartic proteases from marine sponges, which present an abnormal high R f value when run in SDS-PAGE. The special feature of using CTAB is that it binds proteins, making them positively charged and thus migrating in the opposite direction compared to SDS-PAGE.
Assuntos
Ácido Aspártico Endopeptidases/análise , Compostos de Cetrimônio/química , Detergentes/química , Eletroforese em Gel de Poliacrilamida/métodos , Poríferos/enzimologia , Animais , Cetrimônio , Ensaios Enzimáticos/métodos , Concentração de Íons de Hidrogênio , Poríferos/químicaRESUMO
The properties and photochemical and photophysical behavior of the mycosporine-like amino acids (MAAs) shinorine and porphyra-334 were experimentally evaluated in solutions of direct ionic micelles as simple biomimicking environments. The preferential partition of the natural molecules in the aqueous phase of sodium dodecyl sulfate (SDS) or cetyltrimethylammonium chloride (CTAC) micellar systems is confirmed. Although the proton dissociation of the carboxylic groups in the MAAs is slightly inhibited in CTAC solutions, the molecules are predicted to be in the form of zwitterions in all the explored media around physiological pH. The increase in the fluorescence quantum yield, emission lifetime and stationary anisotropy in the presence of CTAC micelles suggest electrostatic attractions of the MAAs with the surface of the cationic micelles. Consistently, the triplet-triplet absorption spectra in CTAC solutions reveal the typical environmental features of the micellar interface, while in the presence of SDS they are similar to those determined in neat water. Finally, the photostability of the MAAs increases in the micellar systems, more noticeably in the case of CTAC. It is concluded that the ability of the two MAAs to act as UV screens is susceptible to the influence of electrostatic interactions with organized microheterogeneous environments.
Assuntos
Compostos de Cetrimônio/química , Cicloexanonas/química , Cicloexilaminas/química , Glicina/análogos & derivados , Micelas , Dodecilsulfato de Sódio/química , Cetrimônio , Fluorescência , Glicina/química , Processos Fotoquímicos , Soluções , Raios UltravioletaRESUMO
Flavonoids are polyphenols that help the maintenance of health, aiding the prevention of diseases. In this work, CdTe QDs coated with 3-mercaptopropionic acid (3MPA), with an average size of 2.7nm, were used as photoluminescence probe for flavonoids in different conditions. The interaction between 14 flavonoids and QDs was evaluated in aqueous dispersions in the absence and in the presence of cetyltrimethylammonium bromide (CTAB). To establish a relationship between photoluminescence quenching and the concentration of flavonoids, the Stern-Volmer model was used. In the absence of CTAB, the linear ranges for quercetin, morin and rutin were from 5.0×10-6molL-1 to 6.0×10-5molL-1 and from 1.0×10-5molL-1 to 6.0×10-4molL-1 for kaempferol. The sensibility of the Stern-Volmer curves (Ks) indicated that quercetin interacts more strongly with the probe: Ks quercetin>Ks kaempferol>Ks rutin>Ks morin. The conjugation extension in the 3 rings, and the acidic hydroxyl groups (positions 3'and 4') in the B-ring enhanced the interaction with 3MPA-CdTe QDs. The other flavonoids do not interact with the probe at 10-5molL-1 level. In CTAB organized dispersions, Ks 3-hydroxyflavone>Ks 7-hydroxyflavone>Ks flavona>Ks rutin in the range from 1.0×10-6molL-1 to 1.2×10-5molL-1 for flavones and of 1.0×10-6molL-1 to 1.0×10-5molL-1 for rutin. Dynamic light scattering, conductometric measurements and microenvironment polarity studies were employed to elucidate the QDs-flavonoids interaction in systems containing CTAB. The quenching can be attributed to the preferential solubility of hydrophobic flavonoid in the palisade layer of the CTAB aggregates adsorbed on the surface of the 3MPA CdTe QDs.
Assuntos
Ácido 3-Mercaptopropiônico/química , Compostos de Cádmio/química , Compostos de Cetrimônio/química , Flavonoides/química , Pontos Quânticos/química , Telúrio/química , Água/química , Cetrimônio , Luminescência , Micelas , Tamanho da Partícula , Pirenos/química , Soluções , Tensoativos/químicaRESUMO
Gold nanorods (AuNRs) are suitable for constructing self-assembled structures for the development of biosensing devices and are usually obtained in the presence of cetyltrimethylammonium bromide (CTAB). Here, a sulfated chitosan (ChiS) and gum arabic (GA) were employed to encapsulate CTAB/AuNRs with the purpose of studying the interactions of the polysaccharides with CTAB, which is cytotoxic and is responsible for the instability of nanoparticles in buffer solutions. The presence of a variety of functional groups such as the sulfate groups in ChiS and the carboxylic groups in GA, led to efficient interactions with CTAB/AuNRs as evidenced through UV-vis and FTIR spectroscopies. Electron microscopies (HR-SEM and TEM) revealed that nanoparticle clusters were formed in the GA-AuNRs sample, whereas individual AuNRs, surrounded by a dense layer of polysaccharides, were observed in the ChiS-AuNRs sample. Therefore, the presented work contributes to the understanding of the driving forces that control the surface interactions of the studied materials, providing useful information in the building-up of gold self-assembled nanostructures.
Assuntos
Compostos de Cetrimônio/química , Quitosana/química , Ouro/química , Goma Arábica/química , Nanotubos/química , Cetrimônio , Nanotubos/ultraestrutura , Espectrofotometria Ultravioleta , Espectroscopia de Infravermelho com Transformada de FourierRESUMO
Bentonite clays exhibit high adsorptive capacity for contaminants, including aflatoxin B1 (AFB1), a mycotoxin responsible for causing severe toxicity in several species including pigs, poultry and man. Organophilic treatments is known to increase the adsorption capacity of bentonites, and the primary aim of this study was to evaluate the ability of Brazilian bentonite and two organic salts - benzalkonium chloride (BAC) and cetyltrimethylammonium bromide (CTAB) to adsorb AFB1. For this end, 2(2) factorial designs were used in order to analyze if BAC or CTAB was able to increase AFB1 adsorption when submitted in different temperature and concentration. Both BAC and CTAB treatment (at 30°C and 2% of salt concentration) were found to increase the adsorption of AFB1 significantly compared with untreated bentonite. After organophilic bentonite treatments with BAC or CTAB, a vibration of CH stretch (2850 and 2920cm(-1)) were detected. A frequency of the SiO stretch (1020 and 1090cm(-1)) was changed by intercalation of organic cation. Furthermore, the interlayer spacing of bentonite increases to 1.23nm (d001 reflection at 2θ=7.16) and 1.22 (d001 reflection at 2θ=7.22) after the addition of BAC and CTAB, respectively. Another aim of the study was to observe the effects of these two bentonite salts in neural crest stem cell cultures. The two materials that were created by organophilic treatments were not found to be toxic to stem cells. Furthermore the results indicate that the two materials tested may protect the neural crest stem cells against damage caused by AFB1.
Assuntos
Aflatoxina B1/toxicidade , Bentonita/farmacologia , Citoproteção/efeitos dos fármacos , Compostos Orgânicos/química , Células-Tronco/citologia , Adsorção , Análise de Variância , Animais , Compostos de Benzalcônio/química , Cátions , Cetrimônio , Compostos de Cetrimônio/química , Codorniz , Espectroscopia de Infravermelho com Transformada de Fourier , Células-Tronco/efeitos dos fármacos , Temperatura , Vibração , Difração de Raios XRESUMO
Genomic DNA extraction is crucial for molecular research, including diagnostic and genome characterization of different organisms. The aim of this study was to comparatively analyze protocols of DNA extraction based on cell lysis by sarcosyl, cetyltrimethylammonium bromide, and sodium dodecyl sulfate, and to determine the most efficient method applicable to soybean caterpillars. DNA was extracted from specimens of Chrysodeixis includens and Spodoptera eridania using the aforementioned three methods. DNA quantification was performed using spectrophotometry and high molecular weight DNA ladders. The purity of the extracted DNA was determined by calculating the A260/A280 ratio. Cost and time for each DNA extraction method were estimated and analyzed statistically. The amount of DNA extracted by these three methods was sufficient for PCR amplification. The sarcosyl method yielded DNA of higher purity, because it generated a clearer pellet without viscosity, and yielded high quality amplification products of the COI gene I. The sarcosyl method showed lower cost per extraction and did not differ from the other methods with respect to preparation times. Cell lysis by sarcosyl represents the best method for DNA extraction in terms of yield, quality, and cost effectiveness.
Assuntos
Fracionamento Celular/métodos , DNA/química , Spodoptera/química , Animais , Cetrimônio , Compostos de Cetrimônio/química , DNA/genética , Genes de Insetos , Reação em Cadeia da Polimerase/métodos , Dodecilsulfato de Sódio/química , Spodoptera/genéticaRESUMO
In this study, we report a modified CTAB-PVP method combined with silicon dioxide (silica) treatment for the extraction of high quality genomic DNA from a single larva or pupa. This method efficiently obtains DNA from small specimens, which is difficult and challenging because of the small amount of starting tissue. Maceration with liquid nitrogen, phenol treatment, and the ethanol precipitation step are eliminated using this methodology. The A260/A280 absorbance ratios of the isolated DNA were approximately 1.8, suggesting that the DNA is pure and can be used for further molecular analysis. The quality of the isolated DNA permits molecular applications and represents a fast, cheap, and effective alternative method for laboratories with low budgets.