Evaluación de la toxicidad de cobre en suelos a través de biomarcadores de estrés oxidativo en eisenia foetida

Copper toxicity in soil was evaluated using biomarkers of oxidative stress (catalase enzyme activity, superoxide dismutase and lipid peroxidation) in the earthworm Eisenia foetida. Agricultural topsoils from mining areas of the Aconcagua river basin were collected. Total copper concentrations were in the range of 94-959 mg kg-1, while the exchangeable copper concentrations were in the range of 46-2225 µg kg-1. Earthworms exposed to soil with exchangeable copper concentrations above 32 µg kg-1 showed an increase in catalase activity. Parameters of antioxidant activity were more sensitive than the weight change and thus can be used as appropriate biomarkers in Eisenia foetida.

Callus cell culture of Pothomorphe umbellata (L.) under stress condition leads to high content of peroxidase enzyme

Pothomorphe umbellata (L.) known on Brazil as Caapeba has a number of popular medicinal use, and it has been studied in relation to its pharmacological activity. Peroxidase specific activity (units/mg protein) was evaluated in callus cell culture samples of the P.umbellata, grown in two different MS medium (media 1 and media 2), submitted to 16 hours photoperiod or kept in darkness. Cell growth rate curve showed that the best growth indices were observed when media 2 submitted to the photoperiod regime was used, followed by the same media kept in darkness (stress condition). The results obtained also showed that the cell culture grown under stress conditions (darkness) lead to high content of peroxidase enzyme (an increase of 700% was observed). Kinetic constant values of 3.3 mmol.L-1 and 2,8 sec-1 were obtained for kM and v max,, respectively, using guaiacol as enzyme substrate.

MODELING A COMPUTER APPLICATION FOR MANAGEMENT OF MAINTENANCE ACTIVITIES OF UNPAVED ROADS

ABSTRACTThis study presents a contribution to the modeling of a computer application employing a method of serviceability performance for unpaved roads, aiming the management of maintenance/restoration activities of the primary surface layer. The proposed methodology consisted of field inspections during dry (April to September) and rainy (October to March) periods, during which objective evaluations were performed to survey of defects and their densities and degrees of severity. To aid the functional classification of analyzed road sections and the determination of the defect with major influence on the serviceability of these roads, the method of serviceability performance proposed by Silva (2009)was implemented in the Visual Basic for Applications (VBA) language in Microsoft Excel software. With the use of the computer application proposed it was possible to identify among the defects analyzed in field, through the index of serviceability of the sampling unit per defect type (ISUdef), which one had the greatest influence on determining the relative serviceability index per road section (IST). The results allow us to conclude that the computer application Road achieved satisfactory results, since the objective evaluation criteria applied to road sections denotes consistency regarding their serviceability.

Potential of macrophytes for removing atrazine from aqueous solution

The potential of three macrophytes, Azolla caroliniana, Salvinia minima, and Lemna gibba was assessed in this study to select plants for use in environmental remediation contaminated with atrazine. Experiments were carried out in a greenhouse over six days in pots containing Hoagland 0.25 strength nutritive solution at the following atrazine concentrations: 0; 0.01; 0.1; 1.0; 10.0 mg L-1. Decrease in biomass accumulation was observed in the three macrophytes, as well as toxic effects evidenced by the symptomatology developed by the plants which caused their deaths. The chlorosis and necrosis allowed to observe in the plants the high sensitivity of the three species to the herbicide. Plants presented low potential for removal of atrazine in solution when exposed to low concentrations of the herbicide. However, at the 10.0 mg L-1 atrazine concentration, L. gibba and A. caroliniana showed potential to remove the herbicide from the solution (0.016 and 0.018 mg atrazine per fresh mass gram, respectively). This fact likely resulted from the processes of atrazine adsorption by the dead material. The percentage of atrazine removed from the solution by the plants decreased when the plants were exposed to high concentrations of the pollutant. Azolla caroliniana, S. minima, and L. gibba were not effective in removing the herbicide from solution. The use of these species to remedy aquatic environments was shown to be limited.

Anatomy and physiology of Cattail as related to different population densities

The objective of this work was to evaluate the effects of the population density of Typha angustifolia plants in the anatomical and physiological characteristics. Plants were collected from populations of high density (over 50% of colonization capacity) and low density (less than 50% of colonization capacity) and cultivated under controlled greenhouse conditions. Plants from both populations were grown in plastic trays containing 4 L of nutritive solution for 60 days. At the end of this period, the relative growth rate, leaf area ratio, net assimilatory rate, root/shoot ratio, leaf anatomy, root anatomy, and catalase and ascorbate peroxidase activities were evaluated. Plants from high density populations showed increased growth rate and root/shoot ratio. Low density populations showed higher values of stomatal index and density in leaves, as well as increased palisade parenchyma thickness. Root epidermis and exodermis thickness as well as the aerenchyma proportion of high density populations were reduced, these plants also showed increased vascular cylinder proportion. Only catalase activity was modified between the high and low density populations, showing increased values in low density populations. Therefore, different Typha angustifolia plants show differences in its anatomy and physiology related to its origins on high and low density conditions. High density population plants shows increased growth capacity related to lower apoplastic barriers in root and this may be related to increased nutrient uptake capacity.

Changes in Photosynthesis and Oxidative Stress in Wheat Plants Submmited to Herbicides Application

The use of herbicides, even in tolerant crops, can cause stress evidenced by increase phytotoxicity affecting growth and development. The objectives of this study were to evaluate herbicides effect from different mechanisms of action in photosynthetic and oxidative stress parameters, as well visual phytotoxicity and wild radish control in wheat crop, cultivar Quartzo. Two trials were conducted where the first one evaluated the photosynthetic parameters on wheat plants in two seasons collection, following the application of herbicides bentazon, clodinafop, iodosulfuron, metribuzin, metsulfuron and 2,4-D; and the second one evaluated wild radish (Raphanus sativus) control, wheat phytotoxicity and yield due to bentazon, iodosulfuron, metribuzin, metsulfuron and 2,4-D herbicides application. Photosynthetic rate, stomatal conductance and transpiration were negatively affected by metribuzin, metsulfuron and 2,4-D herbicides at 24 and 120 HAS (hours after spraying) compared to control. Oxidative stress was similar or lower to control, when herbicide was applied and, in general, there was no difference between application times. Lipid peroxidation, catalase activity and phenols were higher in the first collection time. The application of herbicides iodosulfuron and 2,4-D reduces chlorophylls and carotenoids in wheat. Herbicides bentazon, iodosulfuron, metribuzin, metsulfuron and 2,4-D are selective to wheat, cultivar Quartzo and do not affect wheat yield. 2,4-D, metribuzin and iodosulfuron are more efficient for wild radish control.

Protective role of antioxidant vitamin E and catechin on idarubicin-induced cardiotoxicity in rats

Idarubicin is an anthracycline antibiotic extensively used in acute leukemia. In the present study we investigated whether vitamin E and catechin can reduce the toxic effects of idarubicin. Vitamin E (200 IU kg-1 week-1), catechin (200 mg kg-1 week-1), idarubicin (5 mg kg-1 week-1), idarubicin + vitamin E (200 IU kg-1 week-1), and idarubicin + catechin (200 mg kg-1 week-1) combinations were given to male Sprague-Dawley rats weighing 210 to 230 g (N = 6/group). Idarubicin-treated animals exhibited a decrease in body and heart weight, a decrease in myocardial contractility, and changes in ECG parameters (P<0.01). Catechin + idarubicin- and vitamin E + idarubicin-treated groups exhibited similar alterations, but changes were attenuated in comparison to those in cardiac muscle of idarubicin-treated rats (P<0.05). Superoxide dismutase and catalase activity was reduced in the idarubicin-treated group (P<0.05). Glutathione peroxidase levels were decreased in the idarubicin-treated group (P<0.05) and reached maximum concentrations in the catechin- and catechin + idarubicin-treated groups compared to control (P<0.01). Malondialdehyde activity was decreased in the catechin + idarubicin-treated groups compared to control and increased in the other groups, reaching maximum concentrations in the vitamin E-treated group (P<0.01). In electron microscopy studies, swelling of the mitochondria and dilatation of the sarcoplasmic reticulum of myocytes were observed in the idarubicin-treated groups. In groups that were given idarubicin + vitamin E and idarubicin + catechin, the only morphological change was a weak dilatation of the sarcoplasmic reticulum. We conclude that catechin and vitamin E significantly reduce idarubicin-induced cardiotoxicity in rats.

Glutathione peroxidase induction protects Saccharomyces cerevisiae sod1deltasod2delta double mutants against oxidative damage

Saccharomyces cerevisiae mutants deficient in superoxide dismutase genes (sod1delta, sod2delta and the double mutant) were subjected to H2O2 stress in the stationary phase. The highest sensitivity was observed in the sod2delta mutant, while the sod1deltasod2delta double mutant was not sensitive. Sod mutants had lower catalase activity (44%) than wild-type cells, independent of H2O2 stress. Untreated cells of sod1deltasod2delta double mutants showed increased glutathione peroxidase activity (126%), while sod1delta had lower activity (77%) than the wild type. Glutathione levels in sod1delta were increased (200-260%) after exposure to various H2O2 concentrations. In addition, the highest malondialdehyde levels could be observed without H2O2 treatment in sod1delta (167%) and sod2delta (225%) mutants. In contrast, the level of malondialdehyde in the sod1deltasod2delta double mutant was indistinguishable from that of the wild type. These results suggest that resistance to H2O2 by sod1deltasod2delta cells depends on the induction of glutathione peroxidase and is independent of catalase, and that glutathione is a primary antioxidant in the defense against H2O2 in stationary phase sod1delta mutants.

Attenuation of phosphamidon-induced oxidative stress and immune dysfunction in rats treated with N-acetylcysteine

The effect of N-acetylcysteine, a thiolic antioxidant, on attenuation of phosphamidon-induced oxidative stress and immune dysfunction was evaluated in adult male Wistar rats weighing 200-250 g. Rats were divided into four groups, 8 animals/group, and treated with phosphamidon, N-acetylcysteine or the combination of both for 28 days. Oral administration of phosphamidon (1.74 mg/kg), an organophosphate insecticide, increased serum malondialdehyde (3.83 ± 0.18 vs 2.91 ± 0.24 nmol/mL; P < 0.05) and decreased erythrocyte superoxide dismutase (567.8 ± 24.36 vs 749.16 ± 102.61 U/gHb; P < 0.05), catalase activity (1.86 ± 0.18 vs 2.43 ± 0.08 U/gHb; P < 0.05) and whole blood glutathione levels (1.25 ± 0.21 vs 2.28 ± 0.08 mg/gHb; P < 0.05) showing phosphamidon-induced oxidative stress. Phosphamidon exposure markedly suppressed humoral immune response as assessed by antibody titer to ovalbumin (4.71 ± 0.51 vs 8.00 ± 0.12 -log2; P < 0.05), and cell-mediated immune response as assessed by leukocyte migration inhibition (25.24 ± 1.04 vs 70.8 ± 1.09%; P < 0.05) and macrophage migration inhibition (20.38 ± 0.99 vs 67.16 ± 5.30%; P < 0.05) response. Phosphamidon exposure decreased IFN-у levels (40.7 ± 3.21 vs 55.84 ± 3.02 pg/mL; P < 0.05) suggesting a profound effect of phosphamidon on cell-mediated immune response. A phosphamidon-induced increase in TNF-α level (64.19 ± 6.0 vs 23.16 ± 4.0 pg/mL; P < 0.05) suggests a contributory role of immunocytes in oxidative stress. Co-administration of N-acetylcysteine (3.5 mmol/kg, orally) with phosphamidon attenuated the adverse effects of phosphamidon. These findings suggest that oral N-acetylcysteine treatment exerts protective effect and attenuates free radical injury and immune dysfunction caused by subchronic phosphamidon exposure.

Development and characterization of orally-disintegrating films for propolis delivery

This study aimed at evaluating the effect of different concentrations of hydrolyzed collagen (HC) on the properties of an orally disintegrating film containing propolis ethanol extract (PEE) as an active component. The films were evaluated in terms of total phenols, mechanical properties, solubility, contact angle, disintegration time, and microstructure. The films were prepared by casting with 2 g of protein mass (gelatin and HC), 30 g of sorbitol/100 g of protein mass, and 100 g of PEE/100 g of protein mass. HC was incorporated at concentrations of 0, 10, 20, and 30 g/100 g of protein mass. It was found that increased concentrations of HC reduced tensile strength and increased elongation; however, all films showed plastic behavior. An increase in solubility at 25 ºC, a reduction in the contact angle, and disintegration time were also observed. Thus, higher concentrations of collagen led to more hydrophilic and more soluble polymeric matrices that showed shorter dissolution time, favoring the use of these materials as carriers for active compounds to be delivered in the oral cavity.

A 28-fold increase in secretory protein synthesis is associated with DNA puff activity in the salivary gland of Bradysia hygida (Diptera, Sciaridae)

When the first group of DNA puffs is active in the salivary gland regions S1 and S3 of Bradysia hygida larvae, there is a large increase in the production and secretion of new salivary proteins demonstrable by [3H]-Leu incorporation. The present study shows that protein separation by SDS-PAGE and detection by fluorography demonstrated that these polypeptides range in molecular mass from about 23 to 100 kDa. Furthermore, these proteins were synthesized mainly in the S1 and S3 salivary gland regions where the DNA puffs C7, C5, C4 and B10 are conspicuous, while in the S2 region protein synthesis was very low. Others have shown that the extent of amplification for DNA sequences that code for mRNA in the DNA puffs C4 and B10 was about 22 and 10 times, respectively. The present data for this group of DNA puffs are consistent with the proposition that gene amplification is necessary to provide some cells with additional gene copies for the production of massive amounts of proteins within a short period of time (Spradling AC and Mahowald AP (1980) Proceedings of the National Academy of Sciences, USA, 77: 1096-1100).

Erythrocyte glucose-6-phosphate dehydrogenase from Brazilian opossum Didelphis marsupialis

In a comparative study of erythrocyte metabolism of vertebrates, the specific activity of glucose-6-phosphate dehydrogenase (G6PD) of the Brazilian opossum Didelphis marsupialis in a hemolysate was shown to be high, 207 ± 38 IU g-1 Hb-1 min-1 at 37ºC, compared to the human erythrocyte activity of 12 ± 2 IU g-1 Hb-1 min-1 at 37ºC. The apparent high specific activity of the mixture led us to investigate the physicochemical properties of the opossum enzyme. We report that reduced glutathione (GSH) in the erythrocytes was only 50% higher than in human erythrocytes, a value lower than expected from the high G6PD activity since GSH is maintained in a reduced state by G6PD activity. The molecular mass, determined by G-200 Sephadex column chromatography at pH 8.0, was 265 kDa, which is essentially the same as that of human G6PD (260 kDa). The Michaelis-Menten constants (Km: 55 µM) for glucose-6-phosphate and nicotinamide adenine dinucleotide phosphate (Km: 3.3 µM) were similar to those of the human enzyme (Km: 50-70 and Km: 2.9-4.4, respectively). A 450-fold purification of the opossum enzyme was achieved and the specific activity of the purified enzyme, 90 IU/mg protein, was actually lower than the 150 IU/mg protein observed for human G6PD. We conclude that G6PD after purification from the hemolysate of D. marsupialis does not have a high specific activity. Thus, it is quite probable that the red cell hyperactivity reported may be explained by increased synthesis of G6PD molecules per unit of hemoglobin or to reduced inactivation in the RBC hemolysate.

Reduced pancreatic β-cell mass is associated with decreased FoxO1 and Erk1/2 protein phosphorylation in low-protein malnourished rats

A low-protein diet leads to functional and structural pancreatic islet alterations, including islet hypotrophy. Insulin-signaling pathways are involved in several adaptive responses by pancreatic islets. We determined the levels of some insulin-signaling proteins related to pancreatic islet function and growth in malnourished rats. Adult male Wistar rats (N = 20 per group) were fed a 17% protein (normal-protein diet; NP) or 6% protein (low-protein diet; LP), for 8 weeks. At the end of this period, blood glucose and serum insulin and albumin levels were measured. The morphometric parameters of the endocrine pancreas and the content of some proteins in islet lysates were determined. The β-cell mass was significantly reduced (≅65%) in normoglycemic but hypoinsulinemic LP rats compared to NP rats. Associated with these alterations, a significant 30% reduction in insulin receptor substrate-1 and a 70% increase in insulin receptor substrate-2 protein content were observed in LP islets compared to NP islets. The phosphorylated serine-threonine protein kinase (pAkt)/Akt protein ratio was similar in LP and NP islets. The phosphorylated forkhead-O1 (pFoxO1)/FoxO1 protein ratio was decreased by 43% in LP islets compared to NP islets (P < 0.05). Finally, the ratio of phosphorylated-extracellular signal-related kinase 1/2 (pErk1/2) to total Erk1/2 protein levels was decreased by 71% in LP islets compared to NP islets (P < 0.05). Therefore, the reduced β-cell mass observed in LP rats is associated with the reduction of phosphorylation in mitogenic-related signals, FoxO1 and Erk proteins. The cause/effect basis of this association remains to be determined.

Growth and plant mass of radish fertirrigated with different doses nitrogen

ABSTRACT The objective of the present study was to evaluate the effect of nitrogen doses applied via fertigation and associated with different types of crop establishment fertilization on growth and biomass of radish. The experiment was conducted in a greenhouse of the Academic Unit of Agricultural Engineering, Federal University of Campina Grande, from April to May 2014. Treatments consisted of five doses of nitrogen fertilizer applied by fertigation (0, 0.7, 1.4, 2.1 and 2.8g per pot) and three types of crop establishment fertilization (humus 2:2; NPK and control), arranged in a 5 x 3 factor design with four repetitions. The 15 treatments were arranged in 60 plots. The nitrogen source used in the study was urea, divided in three applications: the first application was carried out eight days after transplanting, the second, on day 15, and the third, on day 22. The crop establishment fertilization significantly influenced the growth variables and plant mass of the radish on day 35 after transplanting. The highest values of the variables (number of leaves, plant height, bulb diameter, leaf area, fresh mass of the aerial part, dry mass of the aerial part and root/aerial part were observed in the treatment with humus on day 35 after transplanting. The dose of 2.8g nitrogen per pot corresponding to 6.22g of urea per plant provided the highest yield for the variable number of leafs, leaf area and root length on day 35 after transplanting.

Effects of irregular-shift work and physical activity on cardiovascular risk factors in truck drivers

OBJECTIVE: To analyze the putative effect of type of shift and its interaction with leisure-time physical activity on cardiovascular risk factors in truck drivers.METHODS: A cross-sectional study was undertaken on 57 male truck drivers working at a transportation company, of whom 31 worked irregular shifts and 26 worked on the day-shift. Participants recorded their physical activity using the International Physical Activity Questionnaire along with measurements of blood pressure, body mass index and waist-hip ratio. Participants also provided a fasting blood sample for analysis of lipid-related outcomes. Data were analyzed using a factorial model which was covariate-controlled for age, smoking, work demand, control at work and social support.RESULTS: Most of the irregular-shift and day-shift workers worked more than 8 hours per day (67.7% and 73.1%, respectively). The mean duration of experience working the irregular schedule was 15.7 years. Day-shift workers had never engaged in irregular-shift work and had been working as a truck driver for 10.8 years on average. The irregular-shift drivers had lower work demand but less control compared to day-shift drivers (p < 0.05). Moderately-active irregular-shift workers had higher systolic and diastolic arterial pressures (143.7 and 93.2 mmHg, respectively) than moderately-active day-shift workers (116 and 73.3 mmHg, respectively) (p < 0.05) as well as higher total cholesterol concentrations (232.1 and 145 mg/dl, respectively) (p = 0.01). Irrespective of their physical activity, irregular-shift drivers had higher total cholesterol and LDL-cholesterol concentrations (211.8 and 135.7 mg/dl, respectively) than day-shift workers (161.9 and 96.7 mg/dl, respectively (ANCOVA, p < 0.05).CONCLUSIONS: Truck drivers are exposed to cardiovascular risk factors due to the characteristics of the job, such as high work demand, long working hours and time in this profession, regardless of shift type or leisure-time physical activity.