Application of micronucleus test and comet assay to evaluate BTEX biodegradation

The BTEX (benzene, toluene, ethylbenzene and xylene) mixture is an environmental pollutant that has a high potential to contaminate water resources, especially groundwater. The bioremediation process by microorganisms has often been used as a tool for removing BTEX from contaminated sites. The application of biological assays is useful in evaluating the efficiency of bioremediation processes, besides identifying the toxicity of the original contaminants. It also allows identifying the effects of possible metabolites formed during the biodegradation process on test organisms. In this study, we evaluated the genotoxic and mutagenic potential of five different BTEX concentrations in rat hepatoma tissue culture (HTC) cells, using comet and micronucleus assays, before and after biodegradation. A mutagenic effect was observed for the highest concentration tested and for its respective non-biodegraded concentration. Genotoxicity was significant for all non-biodegraded concentrations and not significant for the biodegraded ones. According to our results, we can state that BTEX is mutagenic at concentrations close to its water solubility, and genotoxic even at lower concentrations, differing from some described results reported for the mixture components, when tested individually. Our results suggest a synergistic effect for the mixture and that the biodegradation process is a safe and efficient methodology to be applied at BTEX-contaminated sites. © 2012 Elsevier Ltd.

Analysis of total glucosinolates and chromatographically purified benzylglucosinolate in organic and conventional vegetables

The limited availability of foods that are free of pesticides has led Brazil to search for alternative production methods to meet the desires of consumers. Currently, organic cultivation represents a production system that complies with general expectations of producers and consumers. Organic cultivation is particularly interesting mainly because of its effect on plant secondary metabolite content, which may help plants to naturally combat pests; in humans, these substances can also contribute to the prevention of chronic diseases. We report on the extraction of glucosinolates (both as total glucosinolates and as benzylglucosinolate) with trifluoroacetic acid addition in a 70:30 MeOH:water (v/v). Total glucosinolates, determined by a thioglucosidase coupled assay, were measured in different Brassicaceae species and were similar to values reported in the literature. For broccoli, analyses were carried out separately on inflorescences, leaves and stalks; analyses were also conducted on thermally processed samples to simulate cooking. Furthermore, when the analysis was conducted on conventional and organic products, the highest concentrations of these substances were most often found in organically cultivated Brassicaceae. The benzylglucosinolate concentrations were evaluated on the same samples using HPLC. The concentration of benzylglucosinolate was significantly higher in organically cultivated vegetables, as well. © 2012 Elsevier Ltd.

Pathological and behavioral risk factors for higher serum c-reactive protein concentrations in free-living adults - A Brazilian community-based study

Low-grade chronic systemic inflammation is often associated with chronic non-communicable diseases, and its most frequently used marker, the C-reactive protein (CRP), has become an identifier of such diseases as well as an independent predictor for cardiovascular disorders and mortality. CRP is produced in response to pro-inflammatory signaling and to individual and behavioral factors, leading to pathological states. The aim of this study was to rank the predicting factors of high CRP concentrations in free-living adults from a community-based sample. We evaluated 522 adults (40-84 years old; 381 women) for anthropometric characteristics, dietary intake, clinical and physical tests, and blood analysis. Subjects were assigned to groups, according to CRP concentrations, as normal CRP (G1;<3.0 mg/L; n = 269), high CRP (G2; 3.0-6.0 mg/L; n = 139), and very high CRP (G3; >6.0 mg/dL; n = 116). Statistical comparison between groups used one-way ANOVA or Kruskal-Wallis tests, and prediction of altered values in increasing CRP was evaluated by proportional hazard models (odds ratio). CRP distribution was influenced by gender, body mass index, body and abdominal fatness, blood leukocytes, and neutrophil counts. The higher CRP group was discriminated by the above variables in addition to lower VO2max, serum metabolic syndrome components (triglycerides, glucose, and HDL cholesterol), higher insulin, homeostasis assessment of insulin resistance, uric acid, gamma-GT, and homocysteine. After adjustments, only fatness, blood leukocytes, and hyperglycemia remained as independent predictors for increased serum CRP concentrations. Intervention procedures to treat low-grade chronic inflammation in overweight women would mainly focus on restoring muscle mass and functions in addition to an antioxidant-rich diet. © 2012 Springer Science+Business Media, LLC.

High concentrations of glucose reduce the oxidative metabolism of dog neutrophils in vitro

Background: Dogs are commonly affected by hyperglycemic conditions. Hyperglycemia compromises the immune response and favors bacterial infections; however, reports on the effects of glucose on neutrophil oxidative metabolism and apoptosis are conflicting in humans and rare in dogs. Considering the many complex factors that affect neutrophil oxidative metabolism in vivo, we investigated in vitro the specific effect of high concentrations of glucose on superoxide production and apoptosis rate in neutrophils from healthy dogs.Results: The capacity of the neutrophils to reduce tetrazolium nitroblue decreased significantly in the higher concentration of glucose (15.13 ± 9.73% (8 mmol/L) versus 8.93 ± 5.71% (16 mmol/L)). However, there were no changes in tetrazolium nitroblue reduction at different glucose concentrations when the neutrophils were first activated with phorbol myristate acetate. High concentrations of glucose did not affect the viability and apoptosis rate of canine neutrophils either with or without prior camptothecin stimulation. This study provides the first evidence that high concentrations of glucose inhibit the oxidative metabolism of canine neutrophils in vitro in a manner similar to that which occurs in humans, and that the decrease in superoxide production did not increase the apoptosis rate.Conclusions: A high concentration of glucose reduces the oxidative metabolism of canine neutrophils in vitro. It is likely that glucose at high concentrations rapidly affects membrane receptors responsible for the activation of NADPH oxidase in neutrophils; therefore, the nonspecific immune response can be compromised in dogs with acute and chronic hyperglycemic conditions. © 2013 Bosco et al.; licensee BioMed Central Ltd.

Assessment of the chemopreventive effect of casearin B, a clerodane diterpene extracted from Casearia sylvestris (Salicaceae)

Studies have shown that Casearia sylvestris compounds protect DNA from damage both in vitro and in vivo. Complementarily, the aim of the present study was to assess the chemopreventive effect of casearin B (CASB) against DNA damage using the Ames test, the comet assay and the DCFDA antioxidant assay. The genotoxicity was assessed by the comet assay in HepG2 cells. CASB was genotoxic at concentrations higher than 0.30μM when incubated with the FPG (formamidopyrimidine-DNA glycosylase) enzyme. For the antigenotoxicity comet assay, CASB protected the DNA from damage caused by H2O2 in the HepG2 cell line in concentrations above 0.04μM with post-treatment, and above 0.08μM with pre-treatment. CASB was not mutagenic (Ames test) in TA 98 and TA 102. In the antimutagenicity assays, the compound was a strong inhibitor against aflatoxin B1 (AFB) in TA 98 (>88.8%), whereas it was moderate (42.7-59.4%) inhibitor against mytomicin C (MMC) in TA 102. Additionally, in the antioxidant assay using DCFDA, CASB reduced reactive oxygen species (ROS) generated by H2O2. In conclusion, CASB was genotoxic to HepG2 cells at high concentrations; was protective of DNA at low concentrations, as shown by the Ames test and comet assay; and was also antioxidant. © 2012 Elsevier Ltd.

Physiological Concentrations of Acute-Phase Proteins and Immunoglobulins in Equine Synovial Fluid

Synovial fluid (SF) is capable of reflecting infectious, immunological, or inflammatory joint conditions in horses by altering its composition and appearance. Although plasma and SF compositions are quantitatively different, this latter compartment reflects changes in plasma macromolecules. Therefore, changes in serum immunoglobulin protein concentrations tend also to alter intracapsular levels. Therefore, it is necessary to know the physiological concentrations of proteins present in SF. The aim of this study was to determine the levels of total protein, albumin, transferrin, haptoglobin, α1-acid glycoprotein, ceruloplasmin, and immunoglobulins A and G in SF of six healthy horses. The synovial proteinogram was obtained by sodium dodecyl sulfate polyacrylamide gel electrophoresis. The SF proteins reached a maximum of 25% of serum concentrations, varying inversely with molecular weight of the protein, except for the ceruloplasmin. © 2013 Elsevier Inc.

Concentrations of Cu, Fe, Mn, and Zn in tropical soils amended with sewage sludge and composted sewage sludge

Sewage sludge may be used as an agricultural fertilizer, but the practice has been criticized because sludge may contain trace elements and pathogens. The aim of this study was to compare the effectiveness of total and pseudototal extractants of Cu, Fe, Mn, and Zn, and to compare the results with the bioavailable concentrations of these elements to maize and sugarcane in a soil that was amended with sewage sludge for 13 consecutive years and in a separate soil that was amended a single time with sewage sludge and composted sewage sludge. The 13-year amendment experiment involved 3 rates of sludge (5, 10, and 20 t ha-1). The one-time amendment experiment involved treatments reflecting 50, 100, and 200 % of values stipulated by current legislation. The metal concentrations extracted by aqua regia (AR) were more similar to those obtained by Environmental Protection Agency (EPA) 3052 than to those obtained by EPA3051, and the strongest correlation was observed between pseudo(total) concentrations extracted by AR and EPA3052 and bioavailable concentrations obtained by Mehlich III. An effect of sewage sludge amendment on the concentrations of heavy metals was only observed in samples from the 13-year experiment. © 2012 Springer Science+Business Media B.V.

Mercury concentrations in south atlantic swordfish, Xiphias gladius, Caught off the Coast of Brazil

This study evaluated the concentrations of mercury in fillets (anterior, middle, and end regions) from the swordfish, Xiphias gladius, and the relationships between mercury concentration and fish weight, as well as the region of collection. Of a total of 697 swordfish analyzed, 11 had mercury concentrations above 1 mg/kg, 421 were between 0.5 and 1.0 mg/kg, and 265 were below 0.5 mg/kg. The anterior and posterior regions had greater concentrations of mercury than the middle region, and fish caught off the northern coast of Brazil had a higher concentration than those caught off the southern coast. © 2013 Springer Science+Business Media New York.

Effects of bovine somatotropin injection on serum concentrations of progesterone in non-lactating dairy cows

The objective of this experiment was to evaluate the effects of bovine somatotropin administration on serum concentrations of glucose, insulin, NEFA, IGF-I, and progesterone (P4) in ovariectomized non-lactating dairy cows receiving exogenous P4, as a model to estimate treatment effects on hepatic P4 degradation. Ten non-lactating, non-pregnant, and ovariectomized Gir×Holstein cows were assigned to the experiment (d -14 to 27). On d 0, cows were ranked by BW and BCS, and randomly assigned to one of two treatments: (1) bovine somatotropin (BST; n=5) or (2) saline control (control; n=5). Cows assigned to the BST treatment were administered s.c. injections containing 500. mg of sometribove zinc on d 0, 9, and 18 of the experiment, whereas control cows concurrently received a 10-mL s.c. injection of 0.9% saline. On d -2, cows were inserted with an intravaginal releasing device containing 1.9. g of P4, which remained in the cows until the end the experiment (d 27). Cow BW and BCS were assessed on d -14, 0, and 27. Blood samples were collected daily from d 0 to d 27, at 0 (immediately before), 1, and 2. h relative to concentrate feeding for determination of serum glucose, insulin, NEFA, P4, and IGF-I concentrations. Concentrations of glucose, NEFA, and insulin obtained prior to feeding (0. h) were used to determine pre-prandial revised quantitative insulin sensitivity check index (RQUICKI). No treatment effects were detected for BW (P=0.72) and BCS change (P=0.79) during the experiment. Beginning on d 2 of the experiment, BST cows had greater (P≤0.01) serum IGF-I concentrations compared with control cohorts (treatment×day interaction; P<0.01). Cows receiving BST had greater (P≤0.05) insulin concentrations compared with control cohorts from d 8 to d 11, d 16 and 17, as well as from d 19 to d 21 of the experiment (treatment×day interaction; P<0.01). Cows receiving BST had greater (P≤0.01) mean glucose and NEFA concentrations, as well as reduced (P<0.01) mean RQUICKI during the experiment compared with control cohorts. No treatment effects, however, were detected (P=0.73) for serum P4 concentrations. In conclusion, results from this experiment indicate that hepatic P4 catabolism is not directly regulated by circulating IGF-I, whereas BST administration decreases insulin sensitivity in non-lactating dairy cows in adequate nutritional status. © 2013 Elsevier B.V.

Major components of energy drinks (caffeine, taurine, and guarana) exert cytotoxic effects on human neuronal SH-SY5Y cells by decreasing reactive oxygen species production

Scope. To elucidate the morphological and biochemical in vitro effects exerted by caffeine, taurine, and guarana, alone or in combination, since they are major components in energy drinks (EDs). Methods and Results. On human neuronal SH-SY5Y cells, caffeine (0.125-2 mg/mL), taurine (1-16 mg/mL), and guarana (3.125-50 mg/mL) showed concentration-dependent nonenzymatic antioxidant potential, decreased the basal levels of free radical generation, and reduced both superoxide dismutase (SOD) and catalase (CAT) activities, especially when combined together. However, guarana-treated cells developed signs of neurite degeneration in the form of swellings at various segments in a beaded or pearl chain-like appearance and fragmentation of such neurites at concentrations ranging from 12.5 to 50 mg/mL. Swellings, but not neuritic fragmentation, were detected when cells were treated with 0.5 mg/mL (or higher doses) of caffeine, concentrations that are present in EDs. Cells treated with guarana also showed qualitative signs of apoptosis, including membrane blebbing, cell shrinkage, and cleaved caspase-3 positivity. Flow cytometric analysis confirmed that cells treated with 12.5-50 mg/mL of guarana and its combinations with caffeine and/or taurine underwent apoptosis. Conclusion. Excessive removal of intracellular reactive oxygen species, to nonphysiological levels (or antioxidative stress), could be a cause of in vitro toxicity induced by these drugs. © 2013 Fares Zeidán-Chuliá et al.

Insulin concentrations in cerebellum and body balance in diabetic male rats: Aerobic training effects

Brain insulin has had widespread metabolic, neurotrophic, and neuromodulatory functions and has been involved in the central regulation of food intake and body weight, learning and memory, neuronal development, and neuronal apoptosis. Purpose: The present study investigated the role of swimming training on cerebral metabolism on insulin concentrations in cerebellum and the body balance performance of diabetic rats. Methods: Forty Male Wistar rats were divided in four groups: sedentary control (SC), trained control (TC), sedentary diabetic (SD), and trained diabetic (TD). Diabetes was induced by alloxan (32 mg kg b.w.), single dose injection. The mean blood glucose of diabetic groups was 367 ± 40 mg/dl. Training program consisted in swimming 5 days/week, 1 h/day, 8 weeks, supporting a workload corresponding to 90% of maximal lactate steady state (MLSS). For the body balance testing rats were trained to traverse for 5 min daily for 5-7 days. All dependent variables were analyzed by one-way analysis of variance (ANOVA) and a significance level of p < 0.05 was used for all comparisons. Results: The body balance testing scores were different between groups. Insulin concentrations in cerebellum were not different between groups. Conclusion: It was concluded that in diabetic rats, aerobic training does not induce alterations on cerebellum insulin but induces important metabolic, hormonal and behavioral alterations which are associated with an improvement in glucose homeostasis, serum insulin concentrations and body balance. © 2013 Elsevier Inc.

Diabetes mellitus triggers oxidative stress in the liver of alloxan-treated rats: A mechanism for diabetic chronic liver disease

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Nitrogen and potassium concentrations in the nutrients solution for melon plants growing in coconut fiber without drainage

With the objective of evaluating the effects of N and K concentrations for melon plants, an experiment was carried out from July 1, 2011 to January 3, 2012 in Muzambinho city, Minas Gerais State, Brazil. The Bonus no. 2 was cultivated at the spacing of 1.1 × 0.4. The experimental design was a randomized complete block with three replications in a 4 × 4 factorial scheme with four N concentrations (8, 12, 16, and 20 mmol L-1) and four K concentrations (4, 6, 8, and 10 mmol L-1). The experimental plot constituted of eight plants. It was observed that the leaf levels of N and K, of N-NO3 and of K, and the electrical conductivity (CE) of the substrate increased with the increment of N and K in the nutrients' solution. Substratum pH, in general, was reduced with increments in N concentration and increased with increasing K concentrations in the nutrients' solution. Leaf area increased with increments in N concentration in the nutrients solution. Fertigation with solutions stronger in N (20 mmol L-1) and K (10 mmol L-1) resulted in higher masses for the first (968 g) and the second (951 g) fruits and crop yield (4,425 gm-2). © 2013 Luiz Augusto Gratieri et al.

Effects of dietary clinoptilolite and calcium levels on uric acid and calcium blood concentrations and bone quality of commercial layers

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

Anthropogenic influences on Cd, Cr,Cu, Ni, Pb and Zn concentrations in soils and sediments in a watershed with sugar cane crops at São Paulo State, Brazil

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