Scielo RSS <![CDATA[Brazilian Archives of Biology and Technology]]> vol. 58 num. 4 lang. en <![CDATA[SciELO Logo]]> <![CDATA[Feeding Deterrent and Genotoxicity Analysis of a novel Phytopesticide by using Comet Assay against Helicoverpa armigera (HÜbner) (Lepidoptera: Noctuidae)]]> Newly developed Phytopesticidal formulations from pongam and neem oils were evaluated for their feeding deterrent activity using leaf disc choice and no-choice methods, and genotoxic study using comet assay against Helicoverpa armigera at different concentrations of 5, 10, 15, and 20 ppm. Among various phytopesticidal formulations, neem and pongam oils at 1:1 ratio, called PONNEEM showed significant feeding deterrent activity against H. armigera at 20 ppm concentration and wasgenotoxic to H. armigera (P&gt;0.001). The comet parameters, namely tail moment (arbitrary units), tail length (µm) and tail DNA (%) were observed at all the concentrations of PONNEEM. Statistically significant changes in all the comet parameters of H. armigera were observed at 20 ppm (P&lt;0.001). Feeding deterrent and genotoxicity effect of PONNEEM could be applied as phytopesticide for controlling the lepidopteran insect pests. <![CDATA[Anti-obesity Effects of the Administration of Tournefortia paniculata Cham Extract on Wistar Rats Subjected to a Hypercaloric Diet]]> The aim of this study was to evaluate the therapeutic and toxicologic effects of the administration of the powered vegetable extract and aqueous extract of Tournefortia paniculata leaves on Wistar rats, subjected to a hypercaloric diet for 42 days. The rats were divided into five groups and were given the following treatments by gavage: T0 (control) - 1.0 mL water day-1; T1 - aqueous extract containing 14 mg phenolic compounds kg-1 rat day-1; T2 - 14 mg quercetin kg-1 rat day-1; T3 - 50 mg powered vegetable extract from T. paniculata leaves (PVE) kg-1 rat day-1 and T4 - 100 mg PVE kg-1 rat day-1. The treatments did not significantly alter the weight, but were effective in reducing liver fat, glucose and serum triglycerides. The treatment T1 reduced food consumption and lipid peroxidation. None of the treatments showed genotoxic potential. Results showed that T. paniculata leaves possessed an anti-obesity potential. However, a more detailed study of the medicinal potential and characterization of phytochemicals in this plant would be still necessary for a better understanding of its mechanisms of action, enabling future applications in the treatment of this pathology or for various therapeutic purposes. <![CDATA[Combination vitamin C and vitamin E prevents enteric diabetic neuropathy in the small intestine in rats]]> The present study evaluated the effects of supplementation with a combination of vitamin C and vitamin E on NADH-diaphorase-positive (NADH-d+) and neuronal nitric oxide synthase (nNOS)-immunoreactive myenteric neurons in the duodenum and ileum in diabetic rats. Forty rats were distributed into the following groups: normoglycemic (N), normoglycemic supplemented with vitamin C and vitamin E (NS), diabetic (D), and diabetic supplemented with vitamin C and vitamin E (DS). Vitamin C was added to the drinking water, and vitamin E was incorporated in the diet (1%). After 120 days, the animals were euthanized, and the duodenum and ileum were subjected to NADH-d and nNOS staining. Quantitative and morphometric analyses of myenteric neurons were performed. Diabetes reduced NADH-d+ neurons in the D group. The density of nitrergic neurons was not changed by diabetes or vitamin treatment. Hypertrophy of the cell body area of NADH-d+ and nNOS-immunoreactive neurons was observed in both intestinal segments. Combined supplementation with vitamin C and vitamin E prevented the reduction of the density of NADH-d+ neurons and hypertrophy, demonstratred by both techniques. Supplementation with a combination of vitamin C and vitamin E promoted myenteric neuroprotection in the small intestine in diabetic rats. <![CDATA[Composition and significance of glycosaminoglycans in the uterus and placenta of mammals]]> Glycosaminoglycans (GAGs) are heteropolysaccharides in mammalian tissue and consist of repeated disaccharide units with mono-sulfated or non-sulfated monosaccharides. GAGs are important components of the Extracellular Matrix (ECM) with several physiological roles, in the recognition, migration, adhesion, proliferation and differentiation processes. They are also important in angiogenesis, blood homeostasis, immune reactions, follicule development and also in the development of pathologies such as infertility, tumors and metastases. It has been shown that the profile of glycosaminoglycans in the uterine and placental tissues is highly variable throughout the reproductive cycle and during pregnancy. It may be directly related to their physiological or pathological functions in the tissue. The latter has recently triggered special clinical interest. Current review collaborates for a deeper knowledge on the profile and importance of GAGs in uterine and placental tissues throughout the reproductive cycle and pregnancy. It also covers information on the involvement of these molecules in pathological processes. <![CDATA[Effect of Fibroblast Growth Factor 2 (FGF2) and Insulin Transferrin Selenium (ITS) on In Vitro Maturation, Fertilization and Embryo Development in Sheep]]> The present study evaluated the effect of fibroblast growth factor 2 (FGF2) and insulin-transferrin-selenium (ITS) to the in vitro maturation and embryo culture media on ovine oocyte maturation, cleavage and embryo development. Oocytes having more than five layers of unexpanded cumulus cells and granular homogenous ooplasm were cultured into 50 μL droplets of eight different culture systems: (i) TCM-199 (Tissue Culture Medium-199); (ii) TCM-199+10 ng/mL FGF2; (iii) TCM-199+20 ng/mL FGF2; (iv) TCM-199+30 ng/mL FGF2; (v) TCM-199+10 ng/mL ITS; (vi) TCM-199+20 ng/mL ITS; (vii) TCM-199+30 ng/mL ITS and (viii) TCM-199+20 ng/mL ITS+20 ng/mL FGF2 in a CO2 incubator at 38.50C for 24 h. All the oocyte culture media were supplemented with 10% FBS, FSH (10 μg/mL) and gentamicin (50 µg/mL). The maturation rate was assessed based on the degree of expansion of cumulus cells and identifying first polar body extrusion into perivitelline space. The matured oocytes were inseminated with 1 to 2 million spermatozoa/mL in Brackett and Oliphant medium and the cleavage rate was checked after 42-48 h post insemination and further cultured for 6-7 days. Maturation and cleavage rates were significantly higher (P&lt;0.05) in the oocytes cultured in TCM-199 +10% FBS+FSH (10 μg/mL) supplemented with both 20 ng/mL ITS and 20 ng/mL FGF2 as compared to the control. It was concluded that the supplementation of ITS and FGF2 in maturation medium was beneficial for improving maturation and cleavage rates of sheep oocytes. The addition of ITS and FGF2 in embryo culture medium did not improve the development of sheep embryos. <![CDATA[Epigenetic Modifications: Therapeutic Potential in Cancer]]> Epigenetic modifications and alterations in chromatin structure and function contribute to the cumulative changes observed as normal cells undergo malignant transformation. These modifications and enzymes (DNA methyltransferases, histone deacetylases, histone methyltransferases, and demethylases) related to them have been deeply studied to develop new drugs, epigenome-targeted therapies and new diagnostic tools. Epigenetic modifiers aim to restore normal epigenetic modification patterns through the inhibition of epigenetic modifier enzymes. Four of them (azacitidine, decitabine, vorinostat and romidepsin) are approved by the U.S. Food and Drug Administration. This article provides an overview about the known functional roles of epigenetic enzymes in cancer development. <![CDATA[Exploring the Anticancer Activity of Grape Seed Extract on Skin Cancer Cell Lines A431]]> In this study, grape seeds were extracted using ethyl acetate and petroleum ether by solvent-solvent extraction method. The phytochemical tests were performed to identify different phytochemical compounds present in the grape seed extract (GSE). Antibacterial activity of the GSE was determined using agar diffusion method against Gram- positive and Gram-negative bacteria. Gas chromatography-mass spectrometry (GC-MS) and Fourier transform infrared spectroscopy (FTIR) analysis was done to identify the presence of bioactive compounds and their functional groups. The GC-MS results revealed a total of four compounds, known to have potent activity against cancer cells, viz, squalene, the most potent compound found in ethyl acetate extract and diethyl phthalate, ethyl-9- cis -11- trans octadecadienoate and (R)-(-)-14,-methyl-8-Hexadecyn-1-ol in petroleum ether extract. Cytotoxic activity of the GSE was observed against skin cancer cell lines A4321 using 3-(4, 5-dimethylthiazol-2-yl)-2-5-diphenyl tetrazolium bromide) MTT assay. The IC50 value of the GSE against A431 skin cancer cell line was 480 µg/mL. This is first such report against A4321 cell lines. The study gives the overall perception about importance of GSE in medicine and nutraceuticals purposes. <![CDATA[Heat Shock Protein Hspa5 Interacts with and Protects Tyrosinase Activity]]> In this study, heat shock protein, Hspa5 was cloned, expressed and purified subsequently confirmed that it interacted with the tyrosinase (TYR) in vitro. Then, using the crystal structure of the homologous protein from the bacteria as a template, a homology model of human TYR was constructed. This model was further applied to investigate the molecular docking with Hspa5. The model showed that the interaction between the TYR and Hspa5 was mainly maintained by some hydrogen bonds in a quite low energy state. The results indicated that TYR was protected in different denaturation conditions by Hspa5. It was concluded that Hspa5 served as a molecular chaperone of TYR, which could help to better understand the molecule regulation mechanism of TRY in many kinds of diseases. <![CDATA[Involvement of transforming growth factor beta-1 (TGFβ1) cytokine and FOXP3 transcription factor genetic polymorphisms in hematological malignancies]]> Hematological malignancies (HM) are a group of neoplastic diseases that arise from hematologic cell lineages. Transforming growth factor beta 1 (TGFβ1) is shown to negatively regulate normal and malignant hematopoiesis and, in immunological context, to promote T cell exhaustion and generation of regulatory T cells, which are shown to be deleterious in cancer, by the induction of transcription factor FOXP3 expression. The present study aimed to evaluate TGFB1 exon-1 rs1800470 and FOXP3 intron-1 rs2232365 polymorphisms in relation to HM susceptibility. DNA was extracted from blood samples of 43 HM patients and 142 neoplasia-free individuals and polymorphisms were analyzed by allelic-specific PCR. Association analysis was assessed by the Odds Ratio (OR) with significance level of 5%. Regarding FOXP3 polymorphism, no significant differences were observed in genotype or allele distribution among the patients and controls. However, there was a positive association between TGFB1 TT genotype and HM susceptibility (OR = 4.07; CI95% = 1.94 - 8.52). In the combined analysis, a positive association was also observed for TGFB1 TT and FOXP3 GG genotypes (OR = 4.00; CI95% = 1.54 - 10.41) in relation to HM susceptibility. Our results indicated promising new markers to be further investigated in hematological malignancies. <![CDATA[Potency Evaluation of Recombinant Human Erythropoietin in Brazil: Assessment of Reproducibility Using a Practical Approach]]> In this study, we compared the results of potency determination of recombinant human erythropoietin (rhEPO) obtained between 2010 and 2012 by the National Institute of Quality Control in Health (INCQS/Fiocruz), i.e., the National Control Laboratory (NCL), and by a manufacturer of rhEPO. In total, 47 different batches of commercially prepared rhEPO (alpha isoform) were analyzed. All results, including those of the control and warning limits, remained within the limits recommended by European Pharmacopoeia (Ph. Eur.). All relative error (RE) values were less than ± 30%, wh ereas most were approximately ± 20%. Applying the Bland-Altman plot, only two of 47 values remained outside the limits of agreement (LA). In addition, agreement of potency determination between INCQS and the manufacturer coefficient of variation of reproducibility (% CVR) was considered satisfactory. Taken together, our results demonstrate (i.) the potency assay of rhEPO performed at INCQS, is standardized and controlled, (ii.) the comparison of our results with those of the manufacturer, revealed an adequate inter-laboratory variation, and (iii.) the critical appraisal proposed here appears to be a feasible tool to assess the reproducibility of biological activity, providing additional information regarding monitoring and production consistency to manufacturers and NCLs. <![CDATA[Protective Effect of Carvacrol Against Oxidative Stress and Heart Injury in Cyclophosphamide-Induced Cardiotoxicity in Rat]]> Possible protective effects of carvacrol (Car) against cyclophosphamide (CP)-induced cardiotoxicity was examined in this study. Experimental groups of the rats were randomly divided into 13 groups,each including seven animals: Group 1 (control) treated with saline; groups 2, 3, and 4 treated with 50, 100, or 150 mg/kg of CP, respectively; group 5 treated with 0.5 mL olive oil; groups 6 and 7 treated with 5.0 and 10 mg/kg of Car, respectively; groups 8, 9, or 10 treated with respective CP plus 5.0 mg/kg of Car; and groups 11, 12, or 13 treated with respective CP plus 10 mg/kg of Car. Serum alanine transaminase (ALT),aspartat transaminase (AST), lactate dehydrogenase (LDH), malondialdehyde (MDA),creatine kinase-MB (CK-MB), total oxidant state (TOS), oxidative stress index (OSI), and levels were high only in the CP groups. There was a dose-dependence on the CP-induced cardiotoxicity. Hemorrhage, inflammatory cell infiltration and the separation of the muscle fibers in the heart tissue supported the biochemical data. With 5.0 and 10 mg/kg Car, there was an important decrease in the CP toxicity and this was related to the oxidative and nitrosative stress in the CP-induced cardiotoxicity. Reduced inflammation and lipid peroxidation in the heart tissue and increase of serum glutathione (GSH) and total antioxidant capacity (TAS) levels were found when carvacrol was applied. Based on these findings, it could be proposed that Car was a strong candidate in preventing the CP-induced cardiotoxicity but further clinical studies should be done in order to verify its application on humans. <![CDATA[Role of Carnosine and Melatonin in Ameliorating Cardiotoxicity of Titanium Dioxide Nanoparticles in the Rats]]> The aim of this work was to study the possible cardiotoxicity of two different doses of 50 nm nano titanium dioxide (n-TiO2) and the possible modulating effects of the use of two natural antioxidants carnosine and melatonin. The results showed that TiO2- NPs produced deleterious effects on rat cardiac tissue as confirmed by the increased levels of serum myoglobin, troponin-T and CK-MB. Increased levels of serum Inflammatory markers represented by the tumor necrosis factor alpha (TNF-α) and Interleukin-6 (IL-6) was also noticed. Caspase3 and IGg were elevated compared to the control group in a dose dependant manner. treatment of the rats with Carnosine or melatonin. along with TiO2- NPs administration significantly improved most of the elevated biochemical markers. It was concluded that the use of Carnosine or melatonin could play a beneficial role against deleterious effects of TiO2- NPs <![CDATA[Vitamin E and Sodium Selenite Against Mercuric Chloride-Induced Lung Toxicity in the Rats]]> The aim of the present study was to elucidate the possible protective role of vitamin E and / or sodium selenite on mercuric chloride-induced oxidative stress and histopathological changes in the lung tissue of the rats. Adult male albino Wistar rats were exposed to mercuric chloride (1.0 mg/kg day) for four weeks. Treatment with mercuric chloride led to oxidative stress by enhancing MDA level and also decreasing superoxide dismutase (SOD), catalase (CAT) glutathione peroxidase (GPx) and glutathione S transferaz (GST) activities. However, mercuric chloride exposure resulted in histopathological changes in the lung tissue in the rats. MDA level and SOD, CAT GPx and GST activities and histopathological changes modulated in concomitantly supplementation of vitamin E (100 mg/kg day) and /or sodium selenite (0.25 mg/kg day) to mercuric chloride-treated groups. <![CDATA[Bioefficacy of flindersine against Helicoverpa armigera Hübner, Spodoptera litura Fabricius, Anopheles stephensis Liston. and Culex quinquefasciatus Say.]]> Flindersine, an alkaloid isolated from Toddalia asiatica, was evaluated for their antifeedant, larvicidal and growth inhibitory activities against Helicoverpa armigera, Spodoptera litura and larvicidal activity against vector mosquitoes Anopheles stephensi and Culex quinquefasciatus. For this, leaf disc no choice method was used for agricultural pests; larvicidal activity was tested on second and fourth instar larvae for mosquitoes at different concentrations. Flindersine showed antifeedant, larvicidal and growth inhibitory activities against H. armigera and S. litura and larvicidal activity against vector mosquitoes An. stephensi and Cx. quinquefasciatus. It showed high regression (R2) values of 0.91 and 0.87 against H. armigera and S. litura, respectively for antifeedant activity. Flindersine exhibited more than 65% larvicidal activity against both the pests with LC50 values of 443.04 and 568.88 ppm and R2 values of 0.87 and 0.90 against H. armigera and S. litura, respectively. The larval and pupal duration of tested insects increased to more than five days at 1000 ppm when compared with the control. The adult emergence was reduced when the concentration of flindersine was increased. At 1000 ppm, no adult emergence was observed in both the pests. Flindersine exhibited 100% larvicidal activity against both the tested mosquitoes at 20 ppm concentration, which showed LC50 values of 2.90, 4.19, 1.68 and 2.71 ppm for 2nd and 4th instar larvae of Cx. quinquefasciatus and An. Stephensi, respectively. High regression values were observed for antifeedant, larvicidal and growth inhibitory activities. Flindersine could be used to develop an ecofriendly pesticide formulation to control the agricultural pests and vector mosquitoes. <![CDATA[Enzymatic Activity in the Gastrointestinal Tract of Pimelodus maculatus (Teleostei, Siluriformes) in Two Neotropical Reservoirs with Different Trophic Conditions]]> Enzymatic activities for digestion of proteins and carbohydrates were compared among three organs of the digestive system of Pimelodus maculatus in two reservoirs with different trophic conditions during the winter of 2006. The aim was to test the hypothesis that enzymatic activity for the digestion of proteins and carbohydrates differed among organs and that such activities differ between the trophic state of the environment. Enzymatic activities were determined through the assays of specificity for trypsin, chymotrypsin and β-glucosidase enzymes. The intestine had higher trypsin-like enzymatic activities compared to the stomach and liver. The highest β-glucosidase activity was found in the liver compared to the stomach and intestine in the oligotrophic reservoir only. Overall, enzymatic activity did not differ between the eutrophic and oligotrophic reservoirs, although the intestinal chymotrypsin was comparatively higher in the eutrophic reservoir and the hepatic β-glucosidase was higher in the oligotrophic reservoir. These findings indicated that most digestive activity occurred in the intestine for P. maculatus, which was probably related to its omnivorous/carnivorous feeding habits. The highest proteolytic activity in the intestine was expected for most fishes, but the high hepatic β-glucosidase in the oligotrophic reservoir was unexpected. The hepatic β-glucosidase as well as the intestinal chymotrypsin-like activity could be considered as the candidates for biomarkers of environmental quality. <![CDATA[Lake Baikal Endemic Sculpins (Cottoidei): A Promising Model to Study Adaptive Plasticity of Blood Cholesterol Metabolism]]> We analyzed the blood lipid spectra in four closely related sculpin (Cottoidei) species endemic to Lake Baikal. These data characterize the Baikal sculpins as a set of model organisms for studying the adaptive plasticity of cholesterol metabolism and also mechanisms of resistance to the development of dyslipidemia and atherosclerosis. <![CDATA[Sperm and Egg Jelly Coat from Sea Urchin Lytechinus variegatus Collected in Rio de Janeiro Contain Distinct Sialic Acid-Rich Polysaccharides]]> This work found the occurrence of a distinct sialic acid-rich polysaccharide in the sperm surface of the sea urchin Lytechinus variegatus, which differed significantly from a similar molecule found in the egg jelly. The sperm polysaccharide extracted by protease digestion was purified using anion exchange chromatography and characterized using agarose gel electrophoresis, gas chromatography/mass spectrometry and NMR spectroscopy. This polysaccharide was highly sulfated and composed almost exclusively of N-acetylneuraminic acid. In contrast, the sialic acid-rich polysaccharide from the egg jelly was composed of N-glycolylneuraminic acid and contains several other hexoses in its structure. This new molecule could help to characterize in further detail the mechanism of fertilization in the sea urchin model system. Sulfated polysaccharides from the jelly coat of sea urchins showed species-specificity in inducing the sperm acrosome reaction, providing an example of a signal transduction event regulated by the sulfated polysaccharide. The new sialic acid-rich polysaccharide found in the sperm head could represent a new molecule involved in the biology of the sea urchin fertilization. <![CDATA[Gas Chromatography - Mass Spectrometry Analysis and Antibacterial Activity of Bluish-Green Pigment from Pseudomonas sp. JJTBVK (KF836502)]]> The present study was conducted for the isolation of potential bacteria from the desert soil, their molecular identification and prediction of restriction sites of the potential isolate using the bioinformatics tools. Production of the metabolites was done by inoculating in nutrient broth of pH 8.6. Metabolite was bluish-green in color; it was extracted and dried by using methanol and used for partial characterization by using GC-MS spectroscopy. Antibacterial activity was performed with the clinical human pathogenic isolates. The bacterium was identified as Pseudomonas sp. JJTBVK on the basis of 16S rRNA sequencing. The sequence was analyzed for the restriction cleavage sites, which showed that the sequence had various restriction sites for different enzymes. Antibacterial activity (MIC) of methanol extract of the bacterial culture broth showed antibacterial activity (MIC), which was 29, 30, 30 and 29 mm for Pseudomonas aeruginosa, Staphylococcus aureus, Bacillus subtilis and Salmonella typhi, respectively. GC-MS analysis of the methanol extract showed the presence of naphth [2,3-B] azet-2 (1H) -one, 1-phenyl-, which was the characteristic compound showing the antibacterial activity. <![CDATA[Production of Inulinase by Free and Immobilized Cells of Penicillium funiculosum p.36]]> The aim of this work was to optimize the growth conditions and continuous production of the enzyme using free and immobilized cells of inulinase by Penicillium funiculosum. The highest yield of enzyme (163.5U/mL) was obtained when the culture was incubated at 27oC and 200 rpm for 96h in a fermentation medium containing both inulin and peptone as sole carbon and nitrogen source, respectively. When the cells of the P. funiculosum were immobilized on different carriers, especially linen fibers, their production ability was successfully maintained for seven successive batches. When the fermentation was carried out using inulin juice prepared from Jerusalem artichoke tubers (in place of pure inulin), inulinase production could be sustained till the second cultivation batch of the P. funiculosum immobilized on linen fibers, yielding 122 U/mL enzyme. Results proved the feasibility of using crude inulin juice as a simple and economic carbon source for the production of inulinase. <![CDATA[Production of Polyhydroxybutyrate by Bacillus axaraqunsis BIPC01 using Petrochemical Wastewater as Carbon Source]]> The aim of this study was to use petrochemical wastewater as the source of carbon for the production of polyhydroxyalkanoates (PHA) in an effort to decrease its cost of production. For this purpose, PHA producing bacteria were isolated from the petrochemical wastewater of Bandar Imam, Iran. The purified colonies were screened for PHA by Sudan Black B and Nile Blue A staining. Among positively stained bacteria, the best PHA producer was selected on the basis of cell growth, PHA content and the monomer composition of PHA. The phenotypic and genotypic identification this isolate showed it to be Bacillus axaraqunsis. The PHA was produced at a cell density of about 9.46 g/l of maximum concentration of 6.33g/l l, corresponding to 66% of cell dry weight. These results showed that B. axaraqunsis BIPC01 could be a potent PHA producer using wastewater for industrial purpose and simultaneously reducing the environmental pollution.