Nile Red Incubation Time Before Reading Fluorescence Greatly Influences the Yeast Neutral Lipids Quantification.

High-throughput screening methodologies to estimate lipid content in oleaginous yeasts use Nile red fluorescence in a given solvent and optimized excitation/emission wavelengths. However, Nile red fluorescence stabilization has been poorly analyzed, and high variability occurs when relative fluorescence is measured immediately or a few minutes after dye addition. The aim of this work was to analyze the fluorescence of Nile red at different incubation times using a variety of solvents and oleaginous/non-oleaginous yeast strains. We showed that fluorescence stabilization occurs between 20 and 30 min, depending on the strain and solvent. Therefore, we suggest that fluorescence measurements should be followed until stabilization, where Relative Fluorescence Units should be considered after stabilization for lipid content estimation.

Influence of dye and nylon fibers on microwave-cured acrylic resin properties / Influência de corante e fibras de nylon nas propriedades de uma resina acrílica polimerizada por micro-ondas

ABSTRACT Objective: The aim of this study is to analyze if both color and nylon fibers have an influence on microwave-cured acrylic resin properties. Methods: Rectangular and disk-shaped specimens were prepared using acrylic resins; medium pink with and without nylon fibers and colorless without nylon fibers. To obtain the rectangular specimens, a stainless steel die was used with the following dimensions: 64 X 10 x 3 mm (± 0.5mm). To obtain disk-shaped specimens, a die 50mm (±0.5mm) in diameter and 0.5mm (±0.05mm) thick was employed. They were randomized to form groups: control (colorless acrylic) and experimental (medium pink, with and without nylon fibers), with each group consisting of ten (10) specimens rectangular in shape and five (5) disk-shaped. They were analyzed in six (6) assays (izod impact strength, n=10; Knoop hardness, n=10; glass transition temperature, n=3; water sorption and solubility, n=5; degree of monomer/polymer conversion, n=1; flexural strength and flexural modulus, n=10). All variables were subjected to the analysis of variance test followed by Tukey's multiple comparison test, at a 5% level of significance. Results: The analysis of the monomer/polymer degree of conversion did not reveal any difference between the three groups of resins (medium pink, with and without nylon fibers and colorless resin); it was approximately 88%. The results did not show significant differences between the groups for each variable (p>0.05). Conclusion: The results showed that color and presence of nylon fibers in acrylic resins did not affect the properties analyzed in this study.

RESUMO Objetivo: Analisar se corantes e fibras de nylon influenciam nas propriedades de uma resina acrílica ativada por energia de micro-ondas. Métodos: Espécimes retangulares e em forma de disco foram confeccionados em resina acrílica rosa médio com e sem fibras de nylon e incolor sem fibras. Para obter os espécimes retangulares, foi utilizada uma matriz de aço inoxidável com as seguintes dimensões: 64 X 10 X 3 (± 0,5mm) e para obter os espécimes em forma de disco, foi utilizada uma matriz com 50mm (± 0,5mm) de diâmetro e 0,5mm (±0,05mm) de espessura. Os mesmos foram randomizados para formar os grupos controle (acrílico incolor sem fibras) e experimentais (rosa médio com e sem fibras), sendo cada grupo composto por dez (10) espécimes retangulares e cinco (5) em forma de disco. Foram os mesmos então submetidos a seis (6) ensaios (resistência ao impacto izod, n=10; dureza knoop, n=10; temperatura de transição vítrea, n=3; sorção e solubilidade, n=5; grau de conversão, n=1; resistência à flexão e módulo de flexão, n=10). Todas as variáveis foram submetidas à análise de variância seguida pelo teste de comparações múltiplas de Tukey, com nível de significância de 5%. Resultados: O grau de conversão foi de 88% para os 3 grupos de resinas e os resultados não mostraram diferença estatisticamente significativa entre os mesmos para cada variável (p>0,05). Conclusão: Os resultados mostraram que o corante e a presença de fibras de nylon em resina acrílica não afetaram as propriedades analisadas nesse estudo.

Bifunctional fluorescent benzimidazo[1,2-α]quinolines for Candida spp. biofilm detection and biocidal activity.

Biofilms provide an ideal environment for protecting the microbial cells from damage caused by humoral and cellular immune system components, promoting resistance, infections and increasing mortality and morbidity of patients in health facilities. In an attempt to provide an innovative solution for preventing contamination in hospital environments, this study evaluated nine structural complementary fluorescent benzimidazo[1,2-α]quinolines as bifunctional agents that both detect and have biocidal activity against yeast biofilms on stainless steel surfaces. The benzimidazoles' staining capability was determined by a fluorescence microscopy study and spraying the substance on yeast biofilm contaminated stainless steel surfaces. Furthermore, their in vitro human leukocyte cytotoxicity was evaluated with trypan blue and their biocidal activity was determined as the minimum inhibitory concentration against Candida tropicalis, C. albicans and C. parapsilosis strains. Moreover, scanning electron micrographs were recorded to study the biocidal activity. This resulted in the identification of 7, which presents all the desired characteristics (such as solubility) and capabilities (staining and biocide activity against all tested biofilm forming yeast strains) at the same time. As such, benzimidazole 7 has the potential to guarantee the use of disinfected medical and surgical instruments in clinical and surgical procedures, consequently, contributing to an increased safety for patients.

The use of epifluorescence microscopy and fluorescent dyes for visualization of Oxyuris equi eggs.

This study presents a new method for visualization of Oxyuris equi eggs collected by means of a clear adhesive tape applied to the perianal region of horses. The obtained results indicate that this protocol permits a quick, easy, clear and selective visualization of Oxyuris equi eggs even in the presence of feces and other material. Another advantage of this method is that it can be used with fluorescent dyes solubilized in water, which will stain biologic material without dissolving or altering the adhesive tape and is also environmentally safe. Other dyes currently used for staining biologic materials use organic solvents, which may be combined with acids or bases in their formulation, preventing their use with the tape method.

A new methodology for the visualization of latent fingermarks on the sticky side of adhesive tapes using novel fluorescent dyes.

Three novel fluorescent dyes were evaluated for the detection of latent fingermarks on different types and colors of adhesive tapes. Compared with the conventional reagents used to reveal latent fingermarks on these surfaces, these new fluorescent dyes have many advantages. They are highly selective to fingermarks, require only a simple procedure, do not need pre- or post-treatment, have high thermal and photochemical stability, are low in cost and use only water as a solvent. In addition, the emitted fluorescence creates a sharp contrast with the fingermark surface, meaning the fingermarks can be clearly visualized and photographed when excited with longwave ultraviolet light (365nm).

Acrylic resin disinfection by peracetic acid and microwave energy / Avaliação da efetividade do ácido peracético e da irradiação por microondas na desinfecção de resina acrílica

Objective: The objective of this study was to evaluate the effectiveness of disinfection methods in microwave and immersion in peracetic acid in heat-cured, self-cured and microwave-cured acrylic resin, contaminated with Candida albicans.Methods: Five specimens were prepared for each type of acrylic resin. All were infected with Candida Albicans, incubated at 37°C for 24 hours. The group which underwent microwave energy was irradiated with a power of 840W for 1 minute and the other group underwent disinfection by soaking of 0.2% peracetic acid for 5 minutes.Results: All samples proved to be contaminated after the incubation period. After the different processes of disinfection, both immersion in 0.2% peracetic acid as microwave irradiation were effective in disinfection of the 3 types of acrylic resins contaminated by Candida Albicans.Conclusion: Concluded that soaking in 0,2% peracetic acid for 5 minutes with microwave irradiation power 840W for 1 minute are effective methods for disinfecting heat-cured acrylic resin, self-cured acrylic resin and microwave-cured acrylic resin, contaminated with Candida Albicans.

Objetivo: Avaliar a eficácia dos métodos de desinfecção em microondas e imersão em ácido peracético em resina acrílica termopolimerizável, autopolimerizável e resina acrílica polimerizada por microondas, contaminadas com Candida albicans.Métodos: Cinco amostras foram preparadas para cada tipo de resina acrílica. Todas foram infectadas com Candida Albicans, incubadas a 37 ° C durante 24 h. O grupo submetido a energia de microondas foi irradiada com uma potência de 840W durante 1 minuto e o outro grupo foi submetido a desinfecção por imersão de ácido peracético a 0,2% durante 5 minutos.Resultados: Todas as amostras mostraram estar contaminadas depois do período de incubação. Depois dos diferentes processos de desinfecção, tanto a imersão em 0,2% de ácido peracético, como a irradiação de microondas foram eficazes na desinfecção dos 3 tipos de resinas acrílicas contaminados por Candida Albicans.Conclusão: Concluiu-se que a imersão em ácido peracético 0,2% durante 5 minutos e a irradiação de microondas com potência de 840W durante 1 minuto são métodos eficazes para a desinfecção de resina acrílica termopolimerizável, resina acrílica autopolimerizável e resina acrílica polimerizada por microondas, contaminados com Candida Albicans.

Application of 6-nitrocoumarin as a substrate for the fluorescent detection of nitroreductase activity in Sporothrix schenckii.

INTRODUCTION: Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. METHODS: Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. RESULTS: All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. CONCLUSION: The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection.

APPLICATION OF 6-NITROCOUMARIN AS A SUBSTRATE FOR THE FLUORESCENT DETECTION OF NITROREDUCTASE ACTIVITY IN Sporothrix schenckii / Aplicação de 6-nitrocumarina como substrato fluorescente para detecção de atividade nitroredutásica em Sporothrix schenckii

Introduction Sporothrix schenckii is a thermal dimorphic pathogenic fungus causing a subcutaneous mycosis, sporotrichosis. Nitrocoumarin represents a fluorogenic substrate class where the microbial nitroreductase activity produces several derivatives, already used in several other enzyme assays. The objective of this study was the analysis of 6-nitrocoumarin (6-NC) as a substrate to study the nitroreductase activity in Sporothrix schenckii. Methods Thirty-five samples of S. schenckii were cultivated for seven, 14 and 21 days at 35 °C in a microculture containing 6-nitrocoumarin or 6-aminocoumarin (6-AC) dissolved in dimethyl sulfoxide or dimethyl sulfoxide as a negative control, for posterior examination under an epifluorescence microscope. The organic layer of the seven, 14 and 21-day cultures was analyzed by means of direct illumination with 365 nm UV light and by means of elution on G silica gel plate with hexane:ethyl acetate 1:4 unveiled with UV light. Results All of the strains showed the presence of 6-AC (yellow fluorescence) and 6-hydroxylaminocoumarin (blue fluorescence) in thin layer chromatography, which explains the green fluorescence observed in the fungus structure. Conclusion The nitroreductase activity is widely distributed in the S. schenckii complex and 6-NC is a fluorogenic substrate of easy access and applicability for the nitroreductase activity detection. .

Introdução Sporothrix schenckii é um fungo dimórfico térmico, agente etiológico de micose subcutânea, a esporotricose. Nitrocumarina representa classe de substratos fluorogênicos em que a atividade nitroredutásica microbiana produz vários derivados, já utilizados em vários outros ensaios enzimáticos. O objetivo deste estudo foi analisar 6-nitrocumarina (6-NC) como substrato para estudo da atividade nitroredutásica em Sporothrix schenckii. Métodos Trinta e cinco isolados de S. schenckii foram cultivados por sete, 14 e 21 dias a 35 °C em um microcultivo contendo 6-nitrocumarina ou 6-aminocumarina (6-AC) solubilizada em dimetilsulfóxido ou dimetilsulfóxido como controle negativo, para posterior análise em microscópio de epifluorescência. A fase orgânica da cultura de sete, 14 e 21 dias foi analisada por meio de iluminação direta com luz UV de 365 nm e por eluição em placas de sílica gel G com hexano:acetato de etila 1:4 e revelada com luz UV. Resultados Todos os isolados mostraram a presença de 6-AC (fluorescência amarela) e 6-hidroxilaminocumarina (fluorescência azul) em cromatografia em camada delgada, que explica a fluorescência verde observada na estrutura dos fungos. Conclusão A atividade nitroredutásica é amplamente distribuída no complexo S. schenckii e 6-NC é um substrato fluorogênico de fácil obtenção e aplicabilidade para detecção da atividade nitroredutásica. .

Synthesis, characterization, and spectroscopic investigation of benzoxazole conjugated Schiff bases.

Two Schiff bases were synthesized by reaction of 2-(4'-aminophenyl)benzoxazole derivatives with 4-N,N-diethylaminobenzaldehyde. UV-visible (UV-vis) and steady-state fluorescence in solution were applied in order to characterize its photophysical behavior. The Schiff bases present absorption in the UV region with fluorescence emission in the blue-green region, with a large Stokes' shift. The UV-vis data indicates that each dye behaves as two different chromophores in solution in the ground state. The fluorescence emission spectra of the dye 5a show that an intramolecular proton transfer (ESIPT) mechanism takes place in the excited state, whereas a twisted internal charge transfer (TICT) state is observed for the dye 5b. Theoretical calculations were performed in order to study the conformation and polarity of the molecules at their ground and excited electronic states. Using density functional theory (DFT) methods at theoretical levels BLYP/Aug-SV(P) for geometry optimizations and B3LYP/6-311++G(2d,p) for single-point energy evaluations, the calculations indicate that the lowest energy conformations are in all cases nonplanar and that the dipole moments of the excited state relaxed structures are much larger than those of the ground state structures, which corroborates the experimental UV-vis absorption results.

A fast and cost-effective methodology for Fonsecaea pedrosoi ATCC46428 staining using ESIPT fluorescent dyes.

The microscopic morphology of Fonsecaea pedrosoi ATCC46428 was observed using two benzazole derivatives, 2-(2'-hydroxyphenyl)benzoxazole and 2-(5'-amino-2'-hydroxyphenyl)benzoxazole, which emit intense fluorescence by a proton transfer mechanism in the electronically excited state (ESIPT). The cell surface could be successfully stained with fluorescent dye solutions of 10 microM-10 mM using two different fast and cost-effective procedures. At these concentrations, any structure or dye crystallization could be observed. Concerning the external microstructural details, only the amino derivative allowed the differentiation between hyphae and conidia. These dyes presented some advantages comparing to commercial dyes, since the stained cells showed high chemical, thermal and photochemical stability during the experiments and also after several months of storage at room temperature and normal light exposition. Procedure 1 presented the advantage to be used when heating can change the chemical or biochemical cell composition. On the other hand Procedure 2 showed to be useful as a routine methodology for cells staining. The results allowed to propose a simple and highly sensitive assay to study the F. pedrosoi micromorphology by epifluorescence microscopy. This methodology can probably be extended for other fungi of clinical interest.

Lipid-core nanocapsules restrained the indomethacin ethyl ester hydrolysis in the gastrointestinal lumen and wall acting as mucoadhesive reservoirs.

The aim of this work was to investigate if the indomethacin ethyl ester (IndOEt) released from lipid-core nanocapsules (NC) is converted into indomethacin (IndOH) in the intestine lumen, intestine wall or after the particles reach the blood stream. NC-IndOEt had monomodal size distribution (242 nm; PDI 0.2) and zeta potential of -11 mV. The everted rat gut sac model showed IndOEt passage of 0.16 micromol m(-2) through the serosal fluid (30 min). From 15 to 120 min, the IndOEt concentrations in the tissue increased from 6.13 to 27.47 micromol m(-2). No IndOH was formed ex vivo. A fluorescent-NC formulation was used to determine the copolymer bioadhesion (0.012 micromol m(-2)). After NC-IndOEt oral administration to rats, IndOEt and IndOH were detected in the gastrointestinal tract (contents and tissues). In the tissues, the IndOEt concentrations decreased from 459 to 5 microg g(-1) after scrapping, demonstrating the NC mucoadhesion. In plasma (peripheric and portal vein), in spleen and liver, exclusively IndOH was detected. In conclusion, after oral dosing of NC-IndOEt, IndOEt is converted into IndOH in the intestinal lumen and wall before reaching the blood stream. The complexity of a living system was not predicted by the ex vivo gut sac model.

In vitro antifungal activity of 2-(2'-hydroxy-5'-aminophenyl)benzoxazole in Candida spp. strains.

The development of azole antifungals has allowed for the treatment of several fungal infections. However, the use of these compounds is restricted because of their hepatotoxicity or because they need to be administered together with other drugs in order to prevent resistance to monotherapy. Benzoxazole derivatives are among the most thriving molecular prototypes for the development of antifungal agents. 2-(2'-hydroxyphenyl) benzoxazoles are versatile molecules that emit fluorescence and have antibacterial, antiviral and antifungal properties. 2-(2'-hydroxy-5'-aminophenyl) benzoxazole (HAMBO) was tested against Candida yeast. The inhibition provided by HAMBO was lower than that of fluconazole, showing low antifungal activity against Candida spp., but equivalent to that of benzoxazoles tested in similar studies. HAMBO showed fungistatic activity against all analysed strains. This class of novel benzoxazole compounds may be used as template to produce better antifungal drugs.

Genotoxicity of aminohydroxynaphthoquinones in bacteria, yeast, and Chinese hamster lung fibroblast cells.

There are few studies on the biological activity of aminohydroxy derivates of 1,4-naphthoquinone (1,4-NQ) on prokaryotic and eukaryotic cells. We determined the mutagenic activity of 5-amino-8-hydroxy-1,4-naphthoquinone (ANQ) and 5-amino-2,8-dihydroxy-1,4-naphthoquinone (ANQ-OH) as compared to the unsubstituted 1,4-NQ in Salmonella/microsome assay. Potential mutagenic and recombinogenic effects and cytotoxicity were analyzed in haploid and diploid cultures of the yeast Saccharomyces cerevisiae. In Salmonella/microsome assay, 1,4-NQ was not mutagenic, whereas aminohydroxynaphthoquinones were weakly mutagenic in TA98 and TA102 strains. In haploid yeast in stationary growth phase (STAT), mutagenic response was only observed for the hom3 locus at the highest dose. In diploid yeast, aminohydroxynaphthoquinones did not induce any recombinogenic events, but 1,4-NQ was shown to be a recombinogenic agent. These results suggest that aminohydroxynaphthoquinones are weak mutagenic agents only in prokaryotic cells. The cytotoxicity of 1,4-NQ in yeast stationary cells was more significant in diploid cells as compared to that observed in haploid cells. However, ANQ and ANQOH were slightly cytotoxic in all treatments. Genotoxicity of these naphthoquinone compounds was also determined in V79 Chinese hamster lung fibroblast cells using standard Comet, as well as modified Comet assay with the bacterial enzymes formamidopyrimidine DNA-glycosylase (FPG) and endonuclease III (ENDOIII). Both 1,4-NQ and ANQ induced pronounced DNA damage in the standard Comet assay. The genotoxic effect of ANQ-OH was observed only at the highest dose. In presence of metabolic activation all substances showed genotoxic effects on V79 cells. Post-treatment of V79 cells with ENDOIII and FPG proteins did not have a significant effect on ANQ-OH-induced oxidative DNA damage as compared to standard alkaline Comet assay. However, all naphthoquinones were genotoxic in V79 cells in the presence of metabolic activation and post-treatment with enzymes, indicating that all compounds induced oxidative DNA damage in V79 cells. Our data suggest that aminohydroxynaphthoquinone pro-oxidant activity, together with their capability of DNA intercalation, have an important role in mutagenic and genotoxic activities.

Physico-chemical characterization of nanocapsule polymeric wall using fluorescent benzazole probes.

Fluorescent polymers were used to prepare innovative formulations with the objective of verifying the chemical composition of the particle/water interface of nanocapsules at a molecular level. The benzazole dyes distinguish between apolar and polar/protic environments. Comparing the fluorescent behavior of benzoxazole-loaded nanocapsules (entrapped dye) with that of fluorescent-polymeric nanocapsules (chemically bound dye), the results indicated that the latter was exposed to a different environment than that to which the entrapped dye was exposed. The polymer in the nanocapsule suspensions is actually at the oil/water interface, interacting with both inner and outer pseudo-phases at the same time. The polymer is restricted at the particle/water interface forming a wall in nanocapsules. The physico-chemical stability of nanocapsules was studied by fluorescence, light scattering, zeta-potential and potentiometry. After 15, 30, 45 and 60 days of preparation different fluorescent behaviors were observed for the benzimidazole physically entrapped in nanocapsules compared to the benzimidazole chemically bound to the polymer wall. This spectrum presented an isoemissive point indicating that only two species were in equilibrium in the medium. The study showed that the water is increasingly interacting with the polymer in the nanocapsule suspensions.

ESIPT-exhibiting protein probes: a sensitive method for rice proteins detection during starch extraction.

The 2-(4'-isothiocyanate-2'-hydroxyphenyl)benzoxazole dye was successfully applied as label of rice proteins during the alkaline extraction of starch. Direct fluorescence measurements were used to observe the presence of proteins labelled in different steps of rice starch extraction. The results were compared to those obtained with the well-known biuret colorimetric test. Whereas the colorimetric test indicates the absence of protein after the third extraction step, the fluorescence emission of the conjugate could be observed in all extraction steps. The separation of different rice proteins could also be observed.

Influência de um ciclo de aquecimento por microondas na microdureza e temperatura de transição vítrea de resinas acrílicas / Influence of a microwave cycle on microhardness and glass transition temperature of some acrylic denture base resin

A proposta deste estudo foi avaliar a influência de um ciclo de aquecimento em forno de microondas, indicado para desinfecção, sobre a microdureza Knoop e a temperatura de transição vitrea (Tg) de três tipos de resinas acrílicas: termopolimerizável (Classico, Art. Classico, jet, Art, Classico, São Paulo, SP) e polimerizada por energia de microondas (Onda-Cryl, Art, Classico, São Paulo, SP). Para o ensaio de microdureza, foram confeccionados quatro corpos-de-prova de cada tipo de resina acrílica , com 2,2 cm de lado e 2 mm de espessura. Após a obtenção das médias de microdureza, os corpos-de-prova foram submetidos ao ciclo de aquecimento em forno de microondas por um minuto a 850W, sendo então novamente avaliados quanto à microdureza. Para a obtenção da Tg (C), determinada por meio da calorimetria exploratória diferencial (DSC), foram confeccionados seis corpos-de-prova para cada tipo de resina acrílica, nas mesmas dimensões anteriores, sendo três utilizados como controle e três submetidos ao ciclo de aquecimento em forno de microondas durante um minuto a 850W. Os dados foram submetidos ao teste t de Student pareado e os Tg, ao teste t de Student não pareado. Os resultados mostraram um aumento significativo na microdureza para a resina polimerizada por energia de microondas e uma diminuição significativa da Tg para a resina acrílica termopolimerizável. É possivel concluir que um ciclo de aquecimento em forno de microondas aumentou a microdureza da resina de microondas testada e diminuiu a Tg da resina termopolimerizável, servindo de alerta para avaliar com maior cuidado a relação custo/benefício da técnica proposta

Synthesis and spectroscopic characterisation of new ESIPT fluorescent protein probes.

Three new benzazole isothiocyanate fluorescent dyes, 2-(4'-isothiocyanate-2'-hydroxyphenyl)benzoxazole, 2-(4'-isothiocyanate-2'-hydroxyphenyl)benzothiazole and 2-(4'-isothiocyanate-2'-hydroxyphenyl)benzimidazole were synthesised, purified until optical purity grade and characterised by spectroscopic techniques. UV/VIS and steady-state fluorescence were also applied to characterise the photophysical behaviour of the dyes. These dyes exhibit an intense fluorescence emission with a large Stokes shift, inherent to the class of benzazoles which relax by the excited state intramolecular proton transfer (ESIPT) mechanism. The dyes were studied for labeling bovine serum albumin (BSA), resulting conjugates BSA-dye with a remarkable photostability under UV/VIS radiation in relation to classical protein labels. The resulting conjugates presented fluorescence in the blue-green region. Direct fluorescence detection of protein-labeled with those dyes after polyacrylamide gel electrophoresis indicates their potential use as fluorescent probes for proteins.

Antimicrobial activity of 1, 4-naphthoquinones by metal complexation

The effect of metal complexation on the antimicrobial activity of 1,4-naphthoquinones was investigated. Nickel-, chromium-, iron-, copper-, and cobalt-containing metal chelates of 5-amino-8- hydroxy-1,4-naphtoquinone (2) and its acyl-derivatives (3-8) were synthesized and characterized, and their antimicrobial activity was evaluated. Data from infrared spectroscopy indicate that naphthoquinones coordinate through oxygen and nitrogen atoms for 2, and through oxygen atoms when ligands were acyl derivatives 3-8. Susceptibility tests for antimicrobial activity showed that 2 and its acyl derivatives were effective on inhibiting the growth of pathogenic bacteria such as Staphylococcus aureus, Streptococcus uberis and Bacillus cereus, but not Gram-negative bacteria. The metal complexation often caused decrease of biological activity.Nickel complex of 2 was the most effective against Gram-positive bacteria, showing MIC values ranging from 375 to 1400 mg/ml. Metal chelates may be useful tools for the understanding of the antimicrobial mechanism of 1,4-naphthoquinones on these bacteria

A new totally flat N(sp(2))C(sp(2))N(sp(2)) pincer palladacycle: synthesis and photoluminescent properties.

The Sonogashira coupling of 2-bromopyridine with 8-quinolinyl-acetylene affords 2-pyridinyl-8-quinolinyl-acetylene (1) in high yields. The chloropalladation of 1 with Li(2)PdCl(4) in methanol at room temperature affords the pincer palladacycle (C(5)H(4)N-2-C=C(Cl)-8-C(9)H(6)kappaN,kappaC,kappaN)PdCl (2) in 63% yield. The X-ray molecular structure of (2) shows that it is totally flat and that it is associated in pairs though pi-stacking between alternate pyridine-quinoline moieties (3.448 A). The pairs are also connected by pi-stacking with an interpair distance of 3.452 A between quinoline-quinoline moieties. A very low fluorescence emission was also revealed by the pincer palladacycle 2 in both solution and the solid state, which has been ascribed to an excimeric emission due to the particular structure (rigid and totally flat) of 2 in the solid state.

Use of 1,4-naphthoquinones for control of Erwinia carotovora.

The antimicrobial effect of 5 naphthoquinones was tested against the phytopathogenic bacteria Erwinia carotovora. Disk diffusion tests and determination of minimal inhibitory concentrations (MIC) indicate that the compound naphthazarin (NTZ) has the best antibacterial activity among the naphthoquinones tested. Studies on the mode of action indicate the effect of NTZ was bactericidal at 10 microg/mL. When cultivation was done in the presence of sodium ascorbate, the restoration of E. carotovora growth was observed with 3 microg/mL NTZ, but not when a 10 microg/mL dose was used. The incubation of NTZ with bacterial suspension of E. carotovora resulted in important changes in the absorption spectra of this naphthoquinone, indicating that a redox reaction takes place. These results may suggest that NTZ induces an increase of reactive oxygen species that are toxic to the cell. The compound NTZ was also effective in preventing E. carotovora growth on potato tubers, inhibiting the soft rot development at a concentration of 2 mg/mL.