A new biophysics approach using photoacoustic spectroscopy to study the DNA-ethidium bromide interaction

We have examined the binding processes of ethidium bromide interacting with calf thymus DNA using photoacoustic spectroscopy. These binding processes are generally investigated by a combination of absorption or fluorescence spectroscopies with hydrodynamic techniques. The employment of photoacoustic spectroscopy for the DNA-ethidium bromide system identified two binding manners for the dye. The presence of two isosbestic points (522 and 498 nm) during DNA titration was evidence of these binding modes. Analysis of the photoacoustic amplitude signal data was performed using the McGhee-von Hippel excluded site model. The binding constant obtained was 3.4 x 10(8) M(bp)(-1), and the number of base pairs excluded to another dye molecule by each bound dye molecule (n) was 2. A DNA drug dissociation process was applied using sodium dodecyl sulfate to elucidate the existence of a second and weaker binding mode. The dissociation constant determined was 0.43 mM, whose inverse value was less than the previously obtained binding constant, demonstrating the existence of the weaker binding mode. The calculated binding constant was adjusted by considering the dissociation constant and its new value was 1.2 x 10(9) M(bp)(-1) and the number of excluded sites was 2.6. Using the photoacoustic technique it is also possible to obtain results regarding the dependence of the quantum yield of the dye on its binding mode. While intercalated between two adjacent base pairs the quantum yield found was 0.87 and when associated with an external site it was 0.04. These results reinforce the presence of these two binding processes and show that photoacoustic spectroscopy is more extensive than commonly applied spectroscopies.

The effect of chilling on the photosynthetic activity in coffee (Coffea arabica L.) seedlings. The protective action of chloroplastid pigments

Coffea arabica is considered to be sensitive to low temperatures, being affected throughout its entire life cycle. Injury caused by chilling (low temperatures above zero degree centigrade) is characterized primarily by inhibition of the photosynthetic process. The objective of this work was to evaluate the role of photosynthetic pigments in the tolerance of coffee (C. arabica L.) seedlings to chilling. The evaluation the photosynthetic activity was made by emission of Chl a fluorescence at room temperature (25°C) in vivo and in situ, using a portable fluorometer. The pigment content was obtained by extraction with 80% acetone, while estimation of membrane lipid peroxidation was determined by measuring the MDA content in leaf tissue extracts. The results indicated a generalized reduction in the quantum yield of PSII when the seedlings were maintained in the dark. The reduction occurred in the seedlings submitted to chilling treatment as well as in the control ones. This demonstrates that not only chilling acts to cause an alteration in PSII. It is possible that the tissue storage reserves had been totally exhausted, with the respiratory rate exceeding the photosynthetic rate; the later was nil, since the seedlings were kept in the dark. The efficiency in the capture, transfer and utilization of light energy in PS11 photochemical reactions requires a sequence of photochemical, biochemical and biophysical events which depend on the structural integrity of the photosynthetic apparatus. However, this efficiency was found to be related to the protective action of chloroplastid pigments, rather than to the concentration of these pigments.

Photoacclimation in three species of freshwater red algae

Three freshwater Rhodophyta species (Audouinella eugenea, A. hermannii and Compsopogon coeruleus) were tested as to their responses (photosynthesis, growth and pigment concentration) to two irradiances (low light, LL, 65 μmol m -2 s-1 and high light, HL, 300 μmol m-2 s-1) and two periods (short time, ST, 4 d, and long time, LT, 28 d). Higher growth rates were consistently observed at LL but significant differences were observed only for A. hermannii. Higher values of photoinhibition at LL were found for the three species, which is consistent with the dynamic photoinhibition as a reversible photoprotective mechanism against high irradiance. Light-induced decreases of effective quantum yield (EQY) were observed in the three species consisting of pronounced decreases from LL to HL. Rapid increases of non-photochemical quenching (NPQ) were observed mainly at LL, indicating energy dissipation by reaction centers. Results revealed distinct photoacclimation strategies to deal with high irradiances: the two Audouinella species had only characteristics of shade-adapted algae: acclimation by changes of size of photosy stem units (PSU) under LT and by PSU number under ST; higher values of the photoinhibition parameter (β) and NPQ, and lower values of EQY at LL; higher recovery capacity of potential quantum yield (PQY) at LL and under ST; highly significant positive correlation of electron transport rate (ETR) with NPQ. In addition, C. coeruleus mixed some characteristics of sun-adapted algae: acclimation by changes of PSU number under LT and by PSU size under ST; higher recovery capacity of EQY than the other two species; weak or no correlation of ETR with NPQ. Thus, these characteristics indicate that C. coeruleus cope with high irradiances more efficiently than the Audouinella species.

Fluorescência da clorofila-a e variação da simetria como ferramentas de investigação de plantas sob estresse

Environmental disturbances in various systems and individuals can result in a tendency for fluctuating asymmetry, which consists of a non-directional deviation from symmetry of a bilateral structure in a normally distributed population. Individuals unable to exhibit plastic responses to developmental disorders that occur at random, either genetic or environmental, may exhibit slight deviations from symmetry in organs with bilateral structure such as leaves. Thus analyzing plants by the morphological pattern of symmetry of the leaves has become an effective tool to describe the developmental of plants under stress conditions. This study aimed to analyze if the pattern of symmetry can be used as a tool to investigate the stress level to which a plant is subjected, using ultraviolet-C radiation (UV-C) as source of stress. We determined the leaf symmetry deviations (sd), the average length (AL) and the maximum width of the lamina (MWL) and the fluorescence parameters of chlorophyll-a in leaves of young plants of G. max subjected to 0, 30, 60, 120 or 240 minutes per week of UV-C during four weeks, submitting the values to ANOVA and Tukey test (p <0.05). The results showed significant reductions in AL and MWL, significant increase in sd in leaves irradiated and a drop in potential quantum yield of photosystem II (PSII) and in the photosynthesis performance index (IP). The analysis of leaf symmetry showed high correlation with the stress level to which plants were submitted, being an important analysis to ascertain the fitness of the plant because it is a noninvasive method and easy to perform.

Avaliação das respostas fotossintéticas de macroalgas lóticas sobre diferentes composições espectrais da radiação incidente

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Respostas de curto período das características fotossintéticas da alga vermelha Batrachospermum delicatulum a temperatura e irradiância

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Recuperação das trocas gasosas e fluorescência da clorofila em plantas de Annona emarginata (Schltdl.) H. Rainer submetidas a estrese hídrico

Pós-graduação em Ciências Biológicas (Botânica) - IBB

Efeito da reidratação nos aspectos ecofisiológicos da Copaifera langsdorffii Desf

Pós-graduação em Ciências Biológicas (Botânica) - IBB

New insights into the structural characteristics of irradiated crotamine

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Materiais híbridos de sílica/orgânico dopados com rodamina-B: estudo da luminescência e preparação de filmes finos para aplicações em fotônica

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Surface modified Mg-doped ZnO QDs for biological imaging

Fundação Amparo à Pesquisa Estado de São Paulo (FAPESP)

Intensidade do estresse hídrico modula a expressão de aquaporinas durante a reidratação em Sorghum bicolor

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Excited-state dynamics of meso-tetrakis(sulfonatophenyl) porphyrin J-aggregates

The excited-state dynamics of free-base meso-tetrakis(sulfonatophenyl) porphyrin J-aggregates obtained by the Z-scan technique in femto- and picosecond time scales, along with UV-Vis spectroscopy and flash photolysis is reported. Besides obtaining the S-1 state lifetime, the discrimination between internal conversion and intersystem crossing nonradiative processes from that state was also possible, and their rates and respective quantum yields were found. The aggregates present reverse saturable absorption at 532 nm for both singlet and triplet excited states. The data shown is important for several applications such as optical limiting, photodynamic therapy and others. (C) 2011 Elsevier B.V. All rights reserved.

Revision of Singlet Quantum Yields in the Catalyzed Decomposition of Cyclic Peroxides

The chemiluminescence of cyclic peroxides activated by oxidizable fluorescent dyes is an example of chemically initiated electron exchange luminescence (CIEEL), which has been used also to explain the efficient bioluminescence of fireflies. Diphenoyl peroxide and dimethyl-1,2-dioxetanone were used as model compounds for the development of this CIEEL mechanism. However, the chemiexcitation efficiency of diphenoyl peroxide was found to be much lower than originally described. In this work, we redetermine the chemiexcitation quantum efficiency of dimethyl-1,2-dioxetanone, a more adequate model for firefly bioluminescence, and found a singlet quantum yield (Phi(s)) of 0.1%, a value at least 2 orders of magnitude lower than previously reported. Furthermore, we synthesized two other 1,2-dioxetanone derivatives and confirm the low chemiexcitation efficiency (Phi(s) < 0.1%) of the intermolecular CIEEL-activated decomposition of this class of cyclic. peroxides. These results are compared with other chemiluminescent reactions, supporting the general trend that intermolecular CIEEL systems are much less efficient in generating singlet excited states than analogous intramolecular processes (Phi(s) approximate to 50%), with the notable exception of the peroxyoxalate reaction (Phi(s) approximate to 60%).

Synthesis, spectroscopic characterization, photochemical and photophysical properties and biological activities of ruthenium complexes with mono- and bi-dentate histamine ligand

The monodentate cis-[Ru(phen)(2)(hist)(2)](2+) 1R and the bidentate cis-[Ru(phen)(2)(hist)](2+) 2A complexes were prepared and characterized using spectroscopic (H-1, (H-1-H-1) COSY and (H-1-C-13) HSQC NMR, UV-vis, luminescence) techniques. The complexes presented absorption and emission in the visible region, as well as a tri-exponential emission decay. The complexes are soluble in aqueous and non-aqueous solution with solubility in a buffer solution of pH 7.4 of 1.14 x 10(-3) mol L-1 for (1R + 2A) and 6.43 x 10(-4) mol L-1 for 2A and lipophilicity measured in an aqueous-octanol solution of -1.14 and -0.96, respectively. Photolysis in the visible region in CH3CN converted the starting complexes into cis-[Ru(phen)(2)(CH3CN)(2)](2+). Histamine photorelease was also observed in pure water and in the presence of BSA (1.0 x 10(-6) mol L-1). The bidentate coordination of the histamine to the ruthenium center in relation to the monodentate coordination increased the photosubstitution quantum yield by a factor of 3. Pharmacological studies showed that the complexes present a moderate inhibition of AChE with an IC50 of 21 mu mol L-1 (referred to risvagtini, IC50 181 mu mol L-1 and galantamine IC50 0.006 mu mol L-1) with no appreciable cytotoxicity toward to the HeLa cells (50% cell viability at 925 mu mol L-1). Cell uptake of the complexes into HeLa cells was detected by fluorescence confocal microscopy. Overall, the observation of a luminescent complex that penetrates the cell wall and has low cytotoxicity, but is reactive photochemically, releasing histamine when irradiated with visible light, are interesting features for application of these complexes as phototherapeutic agents.