Fe3+-SELECTIVE ENHANCED FLUORESCENCE PROBE BASED ON A RHODAMINE DERIVATIVE

A novel Fe3+-selective and turn-on fluorescent probe 1 incorporating a rhodamine fluorophore and quinoline subunit was synthesized. Probe 1 displayed high selectivity for Fe3+ in CH3CN–H2O (95:5 v/v) in the presence of other relevant metal cations. Interaction with Fe3+ in 1:1 stoichiometry could trigger a significant fluorescence enhancement due to the formation of the ring-open form. The fluorescent response images were investigated by a novel Euclidean distance method based on red, green, and blue values. A linear relationship was observed between fluorescence intensity changes and Fe3+ concentrations from 7.3 × 10−7 to 3.6 × 10−5 mol L−1.

Dielectric spectroscopy and thermally stimulated discharge current in PEEK film

The complex permittivity of films of polyether ether ketone (PEEK) has been investigated over a wide range of frequency. There is no relaxation peak in the range of 1Hz to 10(5) Hz but in the low-frequency side (10-4 Hz) there is an evidence of a peak that also can be observed by thermally stimulated discharge current measurements. That peak is related with the glass transition temperature (Tg) of the polymer. The activation energy of the relaxation was found to be 0.44 eV, similar to that of several synthetic polymers. Space charges are important in the conduction mechanism as shown by discharging transient.

Lychnophoric acid from Lychnophora pinaster: a complete and unequivocal assignment by NMR spectroscopy

The investigation of the hexane extract from aerial parts of Lychnophora pinaster provided, besides others substances, the E-isomer of lychnophoric acid, a sesquiterpene derivative previously isolated from L. affinis.

Determination of hydrochlorothiazide in pharmaceutical formulations by diffuse reflectance spectroscopy

This paper describes a method for quantitative spot test analysis of hydrochlorothiazide using diffuse reflectance spectroscopy. The reflectance measurements were performed analyzing the colored compound (l = 585 nm) produced from the reaction between hydrochlorothiazide and p-dimethylaminocinnamaldehyde (PDAC) in acid medium. This reaction occurred on filter paper after heating to 80ºC for 8 minutes. Factorial designs allowed varying multiple reaction factors simultaneously in order to obtain the best reaction conditions. These factors included heating temperature, heating time, acid volume and PDAC volume. The linearity was studied in the range of 3.36x10-2 to 1.01x10-1 mol L-1 with a correlation coefficient of 0.998. The limit of detection was estimated to be 1.32x10² mol L-1. Commercial samples were analyzed using the proposed method and the results were favorably compared with those of the United States Pharmacopeia method, showing that quantitative spot test analysis by diffuse reflectance could be successfully used to determine hydrochlorothiazide in medicines.

A fluorescence probe study of gemini surfactants in aqueous solution: a comparison between n-2-n and n-6-n series of the alkanediyl-a,w-bis (dimethylalkylammonium bromides)

Two series of alkanediyl-a,w-bis (dimethylalkylammonium bromide (n-2-n and n-6-n; n=8, 10,12, and 16) have been synthesized and their micelles properties studied in aqueous solution using pyrene, pyrenecarboxaldehyde (PCA) and 1,8 anilinonaphtalene sulfonic acid sodium salt (ANS) as fluorescent probes. The micelles from these surfactants have been characterized on the basis of the information provided by micelle-solubilized fluorescent probes. The obtained results indicated that the surfactant concentration at which a marked decrease in l max parameter of pyrenecarboxaldehyde (PCA) occurs corresponds to the CMC determined by conductimetric measurements. Changes in the emission spectra of ANS and PCA observed in the submicellar range for both surfactants series (n-2-n and n-6-n) were interpreted as formation of pre-aggregates. It was found that the dimeric surfactants with long spacer (s= 6) form more hydrated aggregates when compared with those formed by the n-2-n and CnTAB surfactants series. This was attributed to a more difficult packing of n-6-n surfactant molecules to form micelles.

Determination of atenolol in pharmaceutical formulations by diffuse reflectance spectroscopy

A simple analytical method for quantification of atenolol in pharmaceutical formulations by diffuse reflectance spectroscopy is described. The method is based on the reaction, on the filter paper surface, between the drug and p-chloranil producing a colored compound. The best reaction conditions were obtained with 20 µL of atenolol solution and 20 µL of p-chloranil. All reflectance measurements were carried out at 550 nm and the linear range was from 1.13x10-2 to 7.88x10-2 mol L-1 (r = 0.9992). The limit of detection was 2.80 x 10-3 mol L-1. The proposed method was successfully applied to analysis of different commercial brands of pharmaceutical formulations and the results obtained by the proposed method were in good agreement with those obtained using the British Pharmacopoeia method.

Chlorophyll fluorescence of tropical tree species in a semi-deciduous forest gap

The characterization of different ecological groups in a forest formation/succession is unclear. To better define the different successional classes, we have to consider ecophysiological aspects, such as the capacity to use or dissipate the light energy available. The main objective of this work was to assess the chlorophyll fluorescence emission of tropical tree species growing in a gap of a semi-deciduous forest. Three species of different ecological groups were selected: Croton floribundus Spreng. (pioneer, P), Astronium graveolens Jacq. (early secondary, Si), and Esenbeckia febrifuga A. Juss. (late secondary, St). The potential (Fv/Fm) and effective (deltaF/Fm') quantum efficiency of photosystem II, apparent electron transport rate (ETR), non-photochemical (qN) and photochemical (qP) quenching of fluorescence were evaluated, using a modulated fluorometer, between 7:30 and 11:00 h. Values of Fv/Fm remained constant in St, decreasing in P and Si after 9:30 h, indicating the occurrence of photoinhibition. Concerning the measurements taken under light conditions (deltaF/Fm', ETR, qP and qN), P and Si showed better photochemical performance, i.e., values of deltaF/Fm', ETR and qP were higher than St when light intensity was increased. Values of qN indicated that P and Si had an increasing tendency of dissipating the excess of energy absorbed by the leaf, whereas the opposite was found for St. The principal component analysis (PCA), considering all evaluated parameters, showed a clear distinction between St, P and Si, with P and Si being closer. The PCA results suggest that chlorophyll fluorescence may be a potential tool to differentiate tree species from distinct successional groups.

Chloroplastid pigment contents and chlorophyll a fluorescence in Amazonian tropical three species

Plants react to changes in light and hydrological conditions in terms of quantity and composition of chloroplastidic pigments, which affects the photosynthetic properties and consequently the accumulation of plant biomass. Thus, the chloroplastidic pigment concentration and chlorophyll a fluorescence of three Amazonian species (Bertholletia excelsa, Carapa guianensis e Dipteryx odorata) were investigated in sun and shade leaves form the tree crown collected during two distinct periods of precipitation (dry and rainy seasons). Pigment contents were determined by spectrophotometry and fluorescence variables were determined using a portable fluorometer. The results demonstrated that the species showed high concentrations of Chl a, Chl b e Chl total during the wet season in relation to the dry season, especially in shade leaves. A higher concentration of carotenoids was found in B. excelsa, when compared with leaves of C. guianensis and D. odorata. In leaves of B. excelsa and D. odorata no significant difference was found in relation to the photochemistry of photosystem II (Fv/Fm) between the wet and dry seasons. In conclusion, the three species react differently to variations in the light and precipitation conditions regarding light capture, aspects that might be considered in the management of forest plantations.

White mold detection in common beans through leaf reflectance spectroscopy

ABSTRACT This study aimed to identify wavelengths based on leaf reflectance (400-1050 nm) to estimate white mold severity in common beans at different seasons. Two experiments were carried out, one during fall and another in winter. Partial Least Squares (PLS) regression was used to establish a set of wavelengths that better estimates the disease severity at a specific date. Therefore, observations were previously divided in two sub-groups. The first one (calibration) was used for model building and the second subgroup for model testing. Error measurements and correlation between measured and predicted values of disease severity index were employed to provide the best wavelengths in both seasons. The average indexes of each experiment were of 5.8% and 7.4%, which is considered low. Spectral bands ranged between blue and green, green and red, and red and infrared, being most sensitive for disease estimation. Beyond the transition ranges, other spectral regions also presented wavelengths with potential to determine the disease severity, such as red, green, and near infrared.

Chlorophyll a fluorescence in rice plants exposed of herbicides of group imidazolinone

The objective of this work was to investigate the injuries caused to the photosynthetic apparatus of three types of rice exposed to application of imidazolinone group herbicides. Two experiments were conducted using herbicides Imazethapyr+imazapic and Imazapyr+imazapic, in a split-plot experimental design, and a 3 x 3 factorial, with six replications. The first factor (A) consisted of the herbicide rates 0, 100 e 200 g ha-1 of Imazethapyr+imazapic and 0, 140 e 280 g ha-1 of Imazapyr+imazapic; factor B consisted of type of rice (cv. Puitá Inta CL, sensitive red rice ecotype and red rice ecotype with suspected herbicide tolerance to Imidazolinone). Chlorophyll a fluorescence parameters were evaluated in plants at 30 days after herbicide application, using a portable fluorometer (HandyPEA, Hanstech). The photosynthetic metabolism of cv. Puitá Inta CL was found to tolerate commercial dosages of both herbicides. High sensitivity to the herbicides was observed for the sensitive red rice ecotype, while the photosynthetic apparatus of red rice ecotype with suspected herbicide tolerance showed high tolerance to both herbicides applied at rates higher than the commercial rate. The application of chemical herbicides of the imidazolinone group on rice plants causes changes in the photosynthetic metabolism of plants, detected by evaluating the emission of transient chlorophyll a fluorescence. This method can be useful in helping detect resistance and/or tolerance of red rice plants to herbicides of the imidazolinone group.

Chlorophyll fluorescence in guanandi tree (Calophyllum brasiliense) after herbicide application

Chlorophyll fluorescence is currently used as a rapid diagnostic and nondestructive method to detect and quantify damage on the photosynthetic apparatus of leaves on weeds, crops and ornamental/coniferous trees in response to both environmental stress and herbicides. This study aimed to evaluate chlorophyll fluorescence in guanandi plants (Calophyllum brasiliense) after application of different postemergence herbicides. The experiment was performed in a completely randomized design, with six treatments (control, bentazon, sulfentrazone, isoxaflutole, atrazine and glyphosate) and five replications. The herbicide treatments were applied with a stationary sprayer, and electron transport rate (ETR) was subsequently analyzed with OS5p Multi-Mode Chlorophyll Fluorometer. In the monitored period, guanandi plants subjected to atrazine showed higher sensitivity to chlorophyll fluorescence than the other treatments. Although bentazon is a photosystem II inhibitor, it showed no major changes in electron transport for the studied species and in the monitored period. In summary, ETR is a good parameter to evaluate the effect of some herbicides on Calophyllum brasiliense plants.

Chlorophyll fluorescence parameters in populations of two legume trees: Stryphnodendron adstringens (Mart.) Coville (Mimosoideae) and Cassia ferruginea (Schrad.) Schrad. ex DC. (Caesalpinoideae)

The aim of this study was to investigate the photosynthetic performance in populations of two legume tree species, Stryphnodendron adstringens (Mimosoideae), typical from Cerrado, and Cassia ferruginea (Caesalpinoideae) from the Atlantic Rain Forest. The photosynthetic traits were assessed by measures of chlorophyll fluorescence in progenies of naturally pollinated plants from three populations of S. adstringens and a population of C. ferruginea. Plants of S. adstringens growing under similar conditions of C. ferruginea plants demanded higher light values for photosynthesis saturation, 600 µmol.m-2.s-1 and 350 µmol.m-2.s-1 respectively, and showed higher intrinsic photosynthetic efficiency of photosystem II, Fv/Fm of 0.814 versus 0.783 in C. ferruginea. The highest values of Fv/Fm observed in S. adstringens can explain the highest electron transport rates (ETR) obtained for this species. No significant differences were found among progenies from different C. ferruginea trees nor among populations of S. adstringens, and only in few cases, variation among progenies within populations were found for S. adstringens plants. The fact that fluorescence parameters distinguished species but not populations or most of progenies may be related to low intraspecific genetic variation of these chlorophyll fluorescence traits or due to lack of expression on genetic differences in plants under no stressful conditions.

Interphase cytogenetics using fluorescence in situ hybridization: an overview of its application to diffuse and solid tissue

Interphase cytogenetics, utilizing fluorescence in situ hybridization (FISH) techniques, has been successfully applied to diffuse and solid tissue specimens. Most studies have been performed on isolated cells, such as blood or bone marrow cells; a few have been performed on cells from body fluids, such as amniotic fluid, urine, sperm, and sputum. Mechanically or chemically disaggregated cells from solid tissues have also been used as single cell suspensions for FISH. Additionally, intact organized tissue samples represented by touch preparations or thin tissue sections have been used, especially in cancer studies. Advantages and pitfalls of application of FISH methodology to each type of specimen and some significant biological findings achieved are illustrated in this overview.

A simple HPLC-fluorescence method for the measurement of R,S-sotalol in the plasma of patients with life-threatening cardiac arrhythmias

R,S-sotalol, a ß-blocker drug with class III antiarrhythmic properties, is prescribed to patients with ventricular, atrial and supraventricular arrhythmias. A simple and sensitive method based on HPLC-fluorescence is described for the quantification of R,S-sotalol racemate in 500 µl of plasma. R,S-sotalol and its internal standard (atenolol) were eluted after 5.9 and 8.5 min, respectively, from a 4-micron C18 reverse-phase column using a mobile phase consisting of 80 mM KH2PO4, pH 4.6, and acetonitrile (95:5, v/v) at a flow rate of 0.5 ml/min with detection at lex = 235 nm and lem = 310 nm, respectively. This method, validated on the basis of R,S-sotalol measurements in spiked blank plasma, presented 20 ng/ml sensitivity, 20-10,000 ng/ml linearity, and 2.9 and 4.8% intra- and interassay precision, respectively. Plasma sotalol concentrations were determined by applying this method to investigate five high-risk patients with atrial fibrillation admitted to the Emergency Service of the Medical School Hospital, who received sotalol, 160 mg po, as loading dose. Blood samples were collected from a peripheral vein at zero, 0.5, 1.0, 1.5, 2.0, 3.0, 4.0, 6.0, 8.0, 12.0 and 24.0 h after drug administration. A two-compartment open model was applied. Data obtained, expressed as mean, were: CMAX = 1230 ng/ml, TMAX = 1.8 h, AUCT = 10645 ng h-1 ml-1, Kab = 1.23 h-1, a = 0.95 h-1, ß = 0.09 h-1, t(1/2)ß = 7.8 h, ClT/F = 3.94 ml min-1 kg-1, and Vd/F = 2.53 l/kg. A good systemic availability and a fast absorption were obtained. Drug distribution was reduced to the same extent in terms of total body clearance when patients and healthy volunteers were compared, and consequently elimination half-life remained unchanged. Thus, the method described in the present study is useful for therapeutic drug monitoring purposes, pharmacokinetic investigation and pharmacokinetic-pharmacodynamic sotalol studies in patients with tachyarrhythmias.

Studies on ATP-diphosphohydrolase nucleotide-binding sites by intrinsic fluorescence

Potato apyrase, a soluble ATP-diphosphohydrolase, was purified to homogeneity from several clonal varieties of Solanum tuberosum. Depending on the source of the enzyme, differences in kinetic and physicochemical properties have been described, which cannot be explained by the amino acid residues present in the active site. In order to understand the different kinetic behavior of the Pimpernel (ATPase/ADPase = 10) and Desirée (ATPase/ADPase = 1) isoenzymes, the nucleotide-binding site of these apyrases was explored using the intrinsic fluorescence of tryptophan. The intrinsic fluorescence of the two apyrases was slightly different. The maximum emission wavelengths of the Desirée and Pimpernel enzymes were 336 and 340 nm, respectively, suggesting small differences in the microenvironment of Trp residues. The Pimpernel enzyme emitted more fluorescence than the Desirée apyrase at the same concentration although both enzymes have the same number of Trp residues. The binding of the nonhydrolyzable substrate analogs decreased the fluorescence emission of both apyrases, indicating the presence of conformational changes in the neighborhood of Trp residues. Experiments with quenchers of different polarities, such as acrylamide, Cs+ and I- indicated the existence of differences in the nucleotide-binding site, as further shown by quenching experiments in the presence of nonhydrolyzable substrate analogs. Differences in the nucleotide-binding site may explain, at least in part, the kinetic differences of the Pimpernel and Desirée isoapyrases.