Chemoprotective activity of the isoflavones, genistein and daidzein on mutagenicity induced by direct and indirect mutagens in cultured HTC cells

Isoflavones are phenolic compounds widely distributed in plants and found in a high percentage in soybeans. They have important biological properties and are regarded as potential chemopreventive agents. The aim of this study was to verify the preventive effect of two soy isoflavones (genistein and daidzein) by a micronucleus assay, analysis of GST activity, and real-time RT-PCR analysis of GSTa2 gene expression. Mutagens of direct (doxorubicin) and indirect (2-aminoanthracene) DNA damage were used. Hepatoma cells (HTC) were treated with genistein or daidzein for 26 h at noncytotoxic concentrations; 10 μM when alone, and 0.1, 1.0 and 10 μM when combined with genotoxic agents. The micronucleus test demonstrated that both isoflavones alone had no genotoxic effect. Genistein showed antimutagenic effects at 10 μM with both direct and indirect DNA damage agents. On phase II enzyme regulation, the current study indicated an increase in total cytoplasmic GST activity in response to genistein and daidzein at 10 μM supplementation. However, the mRNA levels of GSTa2 isozymes were not differentially modulated by genistein or daidzein. The results point to an in vitro antimutagenic activity of genistein against direct and indirect DNA damage-induced mutagenicity. © 2012 Springer Science+Business Media B.V.

Quantification of uncertainties associated with space-time estimates of short-term soil CO2 emissions in a sugar cane area

The characterization of soil CO2 emissions (FCO2) is important for the study of the global carbon cycle. This phenomenon presents great variability in space and time, a characteristic that makes attempts at modeling and forecasting FCO2 challenging. Although spatial estimates have been performed in several studies, the association of these estimates with the uncertainties inherent in the estimation procedures is not considered. This study aimed to evaluate the local, spatial, local-temporal and spatial-temporal uncertainties of short-term FCO2 after harvest period in a sugar cane area. The FCO2 was featured in a sampling grid of 60m×60m containing 127 points with minimum separation distances from 0.5 to 10m between points. The FCO2 was evaluated 7 times within a total period of 10 days. The variability of FCO2 was described by descriptive statistics and variogram modeling. To calculate the uncertainties, 300 realizations made by sequential Gaussian simulation were considered. Local uncertainties were evaluated using the probability values exceeding certain critical thresholds, while the spatial uncertainties considering the probability of regions with high probability values together exceed the adopted limits. Using the daily uncertainties, the local-spatial and spatial-temporal uncertainty (Ftemp) was obtained. The daily and mean emissions showed a variability structure that was described by spherical and Gaussian models. The differences between the daily maps were related to variations in the magnitude of FCO2, covering mean values ranging from 1.28±0.11μmolm-2s-1 (F197) to 1.82±0.07μmolm-2s-1 (F195). The Ftemp showed low spatial uncertainty coupled with high local uncertainty estimates. The average emission showed great spatial uncertainty of the simulated values. The evaluation of uncertainties associated with the knowledge of temporal and spatial variability is an important tool for understanding many phenomena over time, such as the quantification of greenhouse gases or the identification of areas with high crop productivity. © 2013 Elsevier B.V.

Effect of fluoride-treated enamel on indirect cytotoxicity of a16% carbamide peroxide bleaching gel to pulp cells

The aim of this study was to evaluate the possibility of fluoride solutions applied to enamel to protect pulp cells against the trans-enamel and transdentinal cytotoxicity of a 16% carbamide peroxide (CP) bleaching gel. The CP gel was applied to enamel/ dentin discs adapted to artificial pulp chambers (8 h/day) during 1, 7 or 14 days, followed by fluoride (0.05% or 0.2%) application for 1 min. The extracts (culture medium in contact with dentin) were applied to MDPC-23 cells for 1 h, and cell metabolism (MTT assay), alkaline phosphatase (ALP) activity and cell membrane damage (flow cytometry) were analyzed. Knoop microhardness of enamel was also evaluated. Data were analyzed statistically by ANOVA and Kruskal-Wallis tests (a=0.05). For the MTT assay and ALP activity, significant reductions between the control and the bleached groups were observed (p<0.05). No statistically significant difference occurred among bleached groups (p>0.05), regardless of fluoride application or treatment days. Flow cytometry analysis demonstrated 30% of cell membrane damage in all bleached groups. After 14 days of treatment, the fluoride-treated enamel presented significantly higher microhardness values than the bleached-only group (p<0.05). It was concluded that, regardless of the increase in enamel hardness due to the application of fluoride solutions, the treated enamel surface did not prevent the toxic effects caused by the 16% CP gel to odontoblast-like cells.

Estimation of the metabolic equivalent (MET) of an exercise protocol based on indirect calorimetry

Objective: This study aimed to determine the energy expenditure (EE) in terms of caloric cost and metabolic equivalents (METs) of two sessions of an exercise protocol. Methods: Fifteen subjects (51.0 ± 5.5years) performed the exercise sessions (80min), which were composed by (warming, walking and flexibility exercises; Session A) and (warming, walking and local muscular endurance exercises; Session B). Heart hate (HR) was measured during each part of the sessions. In laboratory environment, maximal oxygen consumption (VO2max) and oxygen uptake in rest and exercise conditions (using mean HR obtained in classes) were measured on different days, using indirect calorimetry. Exercise METs were obtained by dividing VO2 in exercise (mL.kg-1.min-1) by VO2 in rest (mL.kg-1.min-1). The EE of the exercises was calculated by the formula: MET x Weight(kg) x Time(min)/60. The results were analyzed by ANOVA with Tuckey post hoc test (p < 0.05). Results: One MET for this group was 2.7 ± 0.1mL.kg-1.min-1. The mean METs of exercises were 4,7 ± 0,8 (warming), 5,8 ± 0,9 (walking) and 3,6 ± 0,7 (flexibility) on session A, and 4,6 ± 1,2 (warming), 5,6 ± 1,0 (walking) and 4.8 ± 1,0 (local muscular endurance exercises) on Session B. The training sessions showed similar energy cost (A: 398 ± 86.72 kcal and B: 404 ± 38.85 kcal; p > 0,05). None of activities were classified into vigorous intensity (> 7 METs). There were no differences on VO2 between walking (15,6 ± 2,8 or 15,4 ± 2,6 mL.kg-1.min-1) and local muscular endurance exercises (13,2 ± 2,9 mL.kg-1.min-1), although both were higher (p > 0.05) than flexibility exercises (10.1 ± 2.2 mL.kg-1.min-1). Conclusion: The proposed protocol achieves the physical activity needed by healthy adults to improve and maintain health, by their structure, moderate intensity, duration, frequency and caloric expenditure.

Indirect measurement of photosynthetic pigments in the leaves of Jatropha curcas

The aim of this work was to generate mathematical models capable of identifying photosynthetic pigments and soluble proteins from the leaves of Jatropha curcas using the relationship between classical readings performed by spectrophotometry and the chlorophyll meter, ClorofiLOG ® 1030. The work was conducted at Embrapa Cotton, in the city of Campina Grande, state of Paraíba, Brazil. For indirect analysis, portable equipment was used to read leaf discs at different stages of development. The chlorophyll in these discs was then determined using a classical method, while the Bradford method was used to determine soluble proteins. The data were subjected to analysis of variance and regression analyses, in which the readings obtained using the portable chlorophyll meter were the dependent variables and the photosynthetic pigments and soluble protein determined by the classical method the independents variables. The results indicated that with the exception of chlorophyll b and soluble protein, the mathematical models obtained with the portable chlorophyll ClorofiLOG ® 1030 can be used to estimate the concentration of photosynthetic pigments with high precision, thus saving time and the chemical reagents required for conventional procedures.

Development of an analytical method for the quantification of pfaffic acid in Brazilian ginseng (Hebanthe eriantha)

Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. © 2013 Elsevier B.V.

Validation of an alternative analytical method for the quantification of antioxidant activity in plant extracts

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Use of algorithms for semi-automatic quantification of pulmonary fibrosis and emphysema

Obtaining a semi-automatic quantification of pathologies found in the lung, through images of high resolution computed tomography (HRCT), is of great importance to aid in medical diagnosis. Paraccocidioidomycosis (PCM) is a systemic disease that affects the lung and even after effective treatment leaves sequels such as pulmonary fibrosis and emphysema. It is very important to the area of tropical diseases that the lung injury be quantified more accurately. In this stud, we propose the development of algorithms in computational environment Matlab® able to objectively quantify lung diseases such as fibrosis and emphysema. The program consists in selecting the region of interest (ROI), and through the use of density masks and filters, obtaining the lesion area quantification in relation to the healthy area of the lung. The proposed method was tested on 15 exams of HRCT of patients with confirmed PCM. To prove the validity and effectiveness of the method, we used a virtual phantom, also developed in this research. © 2013 Springer-Verlag.

Effect of light curing modes on mechanical properties of direct and indirect composites

Objective. To evaluate the degree of conversion (DC), flexural strength (FS) and Knoop microhardness (KHN) of direct and indirect composite resins polymerized with different curing systems. Materials and methods. Specimens of direct (Z250, 3M/Espe) and indirect (Sinfony, 3M/Espe) restorative materials were made and polymerized using two light curing units: XL2500 (3M/Espe) and Visio system (3M/Espe). Absorption spectra of both composites were obtained on a FTIR spectrometer in order to calculate the DC. FS was evaluated in a universal testing machine and surface microhardness was performed in a microhardness tester (50gf/15s). DC, FS and KHN data were submitted to two-way ANOVA and Tukey's test (α = 0.05). Results. Z250 showed higher DC, FS and KHN compared with Sinfony when the polymerization was carried out with XL2500 (p < 0.05). However, there is no statistical difference in DC between the materials when Visio was used (p > 0.05). Visio showed higher DC and KHN for Z250 and Sinfony than the values obtained using XL2500 light curing (p < 0.05). For FS, no significant difference between curing units was found (p > 0.05). Conclusion. Even though the Visio system could increase DC and KHN for some direct and indirect composites, compared with the conventional halogen curing unit, a high number of monomers did not undergo conversion during the polymerization. © 2013 Informa Healthcare.

Yeast proteins originated from the production of brazilian bioethanol quantification and content

The purpose of this work was to determine the levels of protein and the amino acid distribution in the cell mass of yeast strains (Saccharomyces sensu stricto) originated from Brazilian bioethanol industries. The protein was analyzed with the Kjeldahl method and the amino acids, by using high-performance liquid chromatography (HPLC). The percentages of the protein found ranged from 39 to 49%. The results show that in spite of some variation in numbers between the different yeast strains, all of them presented an amino acid profile similar to the one in the literature for S. cerevisae. The amino acids that have occurred in the largest amounts were: aspartic, glutamic acids and lysine, and those in the lowest amounts were: cysteine and methionine. Although the characteristics of the feedstock used and the process conditions are determinant of the protein values obtained in dry mass, this work elucidates that the intrinsic properties of the yeast strain influence these values.

Quantification of methomyl levels in cabbage, tomato, and soya milk using a renewable amperometric biosensor

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

Sensitive voltammetric sensor for fast detection of mycolic acids present in mycobacteria

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

Composition and quantification of the anthropogenic and natural fractions of wastes collected from the stormwater drainage system for discussions about the waste management and people behavior

A study was made of the composition of wastes collected from the pipes of the stormwater drainage system of Sorocaba, SP, Brazil (600 thousand inhabitants). A total of 10 samples weighing at least 100 kg each were sorted into 19 items to determine the fraction that can be considered natural (earth/sand, stones, organic matter, and water, the latter determined after oven-drying the samples) and the anthropogenic fraction (the remaining 15 items, especially construction and demolition wastes and packaging). Soil/sand was found to be the main item collected (52.5 % dry weight), followed by the water soaked into the waste (24.3 %), which meant that all the other wastes were saturated in mud, whose contents varied from 6.4 % (glass) to 87.2 % (metalized plastics packaging). In general, 83 % of the collected wastes can be classified as natural, but the remaining 17 % represent 2,000 kg of the most varied types of wastes discarded improperly every day on the streets of the city. This is an alarming amount of wastes that may clog parts of the drainage systems, causing troubles for all the population (like flooding) and must be strongly considered in municipal solid wastes management and in environmental education programs. © 2013 Springer Science+Business Media Dordrecht.

Indirect georeferencing of terrestrial laser scanning data using control lines

An indirect method for the georeferencing of 3D point clouds obtained with terrestrial laser scanning (TLS) data using control lines is presented. This technique could be used for rapid data acquisition where resources do not permit the use of expensive navigation sensors or the placement of pre-signalised targets. The most important characteristic is the development of a mathematical model based on the principle that the direction vector of the TLS straight line is coplanar with the plane defined by the origin of the TLS system, one endpoint of a control line and the direction vector of the control line in the ground reference coordinate system. The transformation parameters are estimated by minimising the distance between the control lines and their corresponding TLS straight lines. The proposed method was tested using both simulated and real data, and the advantages of this new approach are compared with conventional surveying methods. © 2013 This article is a U.S. Government work and is in the public domain in the USA.

An HPLC-UV method for the quantification of zidovudine in rat plasma

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