Vagner Roberto de Souza

Photodynamic effect of light-emitting diode light on cell growth inhibition induced by methylene blue

The aim of this study was to propose the use of red light-emitting diode (LED) as an alternative light source for methylene blue (MB) photosensitizing effect in photodynamic therapy (PDT). Its effectiveness was tested against Staphylococcus aureus (ATCC 26923), Escherichia coli (ATCC 26922), Candida albicans (ATCC 90028) and Artemia salina. The maximum absorption of the LED lamps was at a wavelength of 663 nm, at intensities of 2, 4, 6 and 12 J.cm−2 for 10, 20, 30 and 60 min of exposure, respectively. Assays with and without LED exposure were carried out in plates containing MB at concentrations of 7 to 140.8 μM for microorganisms and 13.35 to 668.5 μM for microorganisms or microcrustaceans. The LED exposure induced more than 93.05%, 93.7% and 93.33% of growth inhibition for concentrations of 42.2 μM for S. aureus (D-value=12.05 min) and 35.2 μM for E. coli (D-value=11.51 min) and C. albicans (D-value=12.18 min), respectively after 20 min of exposure. LED exposure for 1 h increased the cytotoxic effect of MB against A. salina from 27% to 75%. Red LED is a promising light device for PDT that can effectively inhibit bacteria, yeast and microcrustacean growth.

pKa determinations of xanthene derivates in aqueous solutions by multivariate analysis applied to UV-Vis spectrophotometric data.

Xanthenes form to an important class of dyes which are widely used. Most of them present three acid-base groups: two phenolic sites and one carboxylic site. Therefore, the pKa determination and the attribution of each group to the corresponding pKa value is a very important feature. Attempts to obtain reliable pKa through the potentiometry titration and the electronic absorption spectrophotometry using the first and second orders derivative failed. Due to the close pKa values allied to strong UV-Vis spectral overlap, multivariate analysis, a powerful chemometric method, is applied in this work. The determination was performed for eosin Y, erythrosin B, and bengal rose B, and also for other synthesized derivatives such as 2-(3,6-dihydroxy-9-acridinyl) benzoic acid, 2,4,5,7-tetranitrofluorescein, eosin methyl ester, and erythrosin methyl ester in water. These last two compounds (esters) permitted to attribute the pKa of the phenolic group, which is not easily recognizable for some investigated dyes. Besides the pKa determination, the chemometry allowed for estimating the electronic spectrum of some prevalent protolytic species and the substituents effects evaluation.

O pKa de indicadores ácido-base e os efeitos coloidais

An experiment is proposed that can be included in undergraduate courses of chemistry. The subject is the acidity of organic compounds, which are employed as pH indicators, particularly in acid-base titrations. The indicators used are methyl orange, bromophenol blue and bromocresol green in aqueous medium. The influence of colloidal systems on the equilibrium is evaluated by the pKa. The colloids employed are surfactants like sodium dodecyl sulfate, cetyl-trimethylammonium bromide and a polymeric non-ionic F127 (pluronics). The effect of stabilization promoted by the system on the acidic or basic structureof the indicator establishes the action mechanism of the colloid on the pKa values.

Evaluation of the photodynamic activity of Xanthene Dyes on Artemia salina described by chemometric approaches

The development of drugs for photodynamic therapy (PDT) is an important area of research due to their growing use in medical applications. Therefore, it is important to develop new bioassay methods for PDT photosensitizers that are inexpensive, easy to handle and highly sensitive to environmental conditions. Xanthene dyes (fluorescein, rose bengal B, erythrosine B and eosin Y) with LED light sources were investigated using Artemia salina as a bioindicator of photodynamic activity. In this study, three factors were investigated: (i) photosensitizers concentration, (ii) the LED irradiation time and (iii) the waiting time between the addition of the photosensitizers and the beginning of the irradiation. To analyze the photo-killing of A. salina, it was employed a 2³ full factorial design. The death of A. salina was related to dye structure and the interaction between the irradiation time and the photosensitizers concentration. About 60% of crustaceans death was obtained using rose bengal B, which presentes the highest quantum yield of singlet oxygen due to the number of iodide substituents in the xanthenes ring. The proposed bioassay using A. salina, xanthene dyes and LED irradiation was found suitable for quantitative PDT drug evaluation.

Influences of experimental parameters on the stability of a benzoporphyrin drug in water/ethanol mixtures: a statistical approach investigation

The benzoporphyrin derivative, named B3B, is a promising new drug for application in photodynamic therapy. In aqueous organic solvent mixtures, B3B, can be used in topical application for superficial diseases, like for skin cancer. In this paper, the self-aggregation phenomena of porphyrin molecules in aqueous media was investigated using a chemometric tool to evaluate the effects of experimental variables on the stability of the B3B monomer in water/ethanol mixtures. A 23 factorial design was employed which permitted determination of the magnitude of experimental parameters which influence aggregation of the drug at different pHs using the minimum number of experiments. At first, the pKa values of the B3B in water/ethanol mixtures were determined by spectrophotometric measurements, resulting in pKa1 = 3.3 and pKa2 = 5.7, corresponding to the imino nitrogens and carboxylic groups, respectively. The experimental variables (B3B concentration, time and water/ethanol composition) were evaluated in three protonation states of the drug, at pH 7.0, 4.1 and 2.6. The two levels (low and high) were set up at: time 5 (-) and 360 minutes (+); [B3B] 4.4 × 10-6(-) and 26.5 × 10-6M (+); and water content in ethanol 40 (-) and 60% (+). The main parameter that determines the magnitude of the effects seems to be the charge component of B3B, which can be modulated by the pH. The results from the 23 fractional design indicated that the main variable inducing B3B aggregation is the water percentage in ethanol at each pH selected. The magnitude of this effect was pronounced at pH 4.1 where the carboxylic and nitrogen free base forms of B3B are maintained. The factors of time and [B3B], despite being small, are not insignificant. Finally, the effect of water content was investigated in a wide range of water percentage in these three pHs.

Spectroscopic and electrochemical properties of iron(II) complexes of polydentate Schiff bases containing pyrazine, pyridine and imidazole groups.

We report the synthesis and spectroscopic/electrochemical properties of iron(II) complexes of polydentate Schiff bases generated from 2-acetylpyridine and 1,3-diaminopropane, acetylpyrazine and 1,3-diaminopropane, and from 2-acetylpyridine and L-histidine. The complexes exhibit bis(diimine)-iron(II) chromophores in association with pyrazine, pyridine or imidazole groups displaying contrasting pi-acceptor properties. In spite of their open geometry, their properties are much closer to those of macrocyclic tetraimineiron(II) complexes. An electrochemical/spectroscopic correlation between E degrees (FeIII/II) and the energies of the lowest MLCT band has been observed, reflecting the stabilization of the HOMO levels as a consequence of the increasing backbonding effects in the series of compounds. Mössbauer data have also confirmed the similarities in their electronic structure, as deduced from the spectroscopic and theoretical data.

Self-aggregation processes of 1,6-diphenyl-1,3,5-hexatriene in water/ethanol mixtures with high water percentages.

This work describes the behavior of 1,6-diphenyl-1,3,5-hexatriene (DPH) in ethanol/water mixtures. The dependence of DPH photophysical properties (absorption and fluorescence emission) on the water percentage in ethanol indicates that DPH undergoes self-aggregation processes in solvent conditions above a critical water content. Evidence such as an additional absorption band, Beers law deviation, kinetic behavior, and other experimental results obtained from temperature variation and surfactant addition demonstrated the presence of several types of DPH aggregates. Resonance light scattering measurements proved that the aggregate grew in water-rich media by a self-catalyzed process.

INVESTIGAÇÃO DA SUPRESSÃO DE FLUORESCÊNCIA DE SORO ALBUMINA BOVINA E HUMANA POR COMPLEXO DE RUTÊNIO

The binding of [RuCl2(L)] (L = N,N-bis(7-methyl-2-pyridylmethylene)-1,3-diiminopropane) to bovine and human serum albumin was investigated by the fluorescence quenching technique. The comparison of the quenching effect of serum albumin fluorescence by ruthenium complex allowed the estimation of subdomain IB in BSA and subdomain IIA in HSA as the binding sites for this complex. The results of fluorescence titration revealed that ruthenium complex quenches the intrinsic fluorescence of BSA through a dynamic quenching mechanism, while HSA has a static quenching mechanism. The thermodynamic parameters indicated that hydrophobic forces played a major role in the binding of ruthenium complex to proteins. The process of binding was a spontaneous process in which Gibbs free energy change was negative.

Um aparato para monitorar reações rápidas: um "stopped-flow" artesanal de baixo custo

The modern stopped-flow reaction analyzer has shown high efficiency and flexibility, which provides outstanding sample economy with a dead-time of less than 1 ms. However the cost of the equipment imposes a serious restriction to many Brazilian scientists and teachers. In this work we describe the construction of a low-cost stopped-flow system coupled to a UV-Vis spectrophotometer. The performance of the system was checked by monitoring the kinetics of two reactions: the fading of phenolphthalein in aqueous alkaline solution and the chlorophyll a demetallation in acid medium. The apparatus showed reasonable efficiency with a dead-time of 0.3 to 0.5 s. The very good results obtained in these two illustrative processes show that the system is satisfactory for determining rate constants with mean reaction times ranging from seconds to minutes.