Avaliação in vitro e in vivo da Atividade Antioxidante do Extrato Hidroetanólico de Folhas de Turnera ulmifolia Linn. var. elegans (Turneraceae)

Recently, it has been a increasing interest in the antioxidative role of natural products to aid the endogenous protective biological systems against the deleterious effects of oxygen (ROS) and nitrogen (RNS) reactive species. Many antioxidant compounds, naturally occurring from plant sources. Natural antioxidants can protect and prevent the human body from oxidative stress and retard the progress of many diseases in which free radical are involved. Several plants used in the folk medicine to treat certain disorders that are accompanied by inflammation and other pharmacological properties have been proved their attributed properties, such antioxidant activity. Turnera ulmifolia Linn. var. elegans (Turneraceae), frequently employed by population as a medicinal plant, demonstrated antioxidant activity by in vitro and in vivo assays, using its leaf hydroethanolic extract (10%) he in vitro DPPH radical-scanvenging activity showed a strong antioxidant activity (86.57% ± 0.14), similar to Carduus marianus and catequine effects. For the in vivo assays, adult female Wistar rats (n=48) with carbon tetrachloride hepatic injury induced (2,5mL/kg i.p.) were used, Six groups or rats were uses (n=8) [G1 = control (1,25 mL/kg i.p. vehicle); G2 = CCl4 (2,5 mL/kg i.p.); G3 = CCl4 + extract 7 days (500 mg/kg p.o.); G4 = CCl4 + Legalon® 7 days (50 mg/kg p.o.), G5 = CCl4 + extract 21 days (500 mg/kg p.o.) e G6 = CCl4 + Legalon® 21 days (50 mg/kg p.o.)]. The hepatic oxidative injury was evaluated through biochemical parameters [alanine amino transferase (ALT), aspartate amino transferase (AST)] histopathological study, while thiobarbituric acid reactive products (TBAR), glutathione (GSH), catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) levels were used to evaluate proantioxidant parameters. The plant extract tested was found effective as hepatoprotective as evidenced by a decreasing in the ALT and AST activities (p<0.001) and TBAR (plasma, p<0.001 and liver, p<0.001). Levels of GSH (blood, p<0.001 and liver, p<0.001) and antioxidant enzymes [CAT erythrocyte (p<0.05) and hepatic (p<0.01); SOD erythrocyte (p<0.001) and hepatic (p<0.001); GPx erythrocyte (p<0.001) and hepatic (p<0.001)] were also significantly increased. Histopathological changes induced by CCl4 were significantly reduced by the extract treatment. The data obtained were comparable to that of Legalon®, a reference hepatoprotective drug. The results showed that T. ulmifolia leaf extract protects against CCl4 induced oxidative damage. Therefore, this effect must be associated to its antioxidant activity, attributed to the phenolic compounds, present in these extract, which can act as free radical scavengers

Avaliação da composição química, atividade antioxidante, antibacteriana, antinoceptiva, antiinflamatória e toxicidade do extrato metanólico e frações de folhas de Spondias sp. (Anacardiaceae)

Spondias sp. (Anacardiaceae), popularly known as cajá-umbu, is an endemic plant from Northeastern Brazil, where their leaves are widely used in folk medicine to treat inflammatory processes, while their fruits have a great agro industrial potential. This study was designed to evaluate hepatoprotective, antinociceptive, antioxidant, antimicrobial and anti-inflammatory properties, as well as the acute toxicity and repeated dose 28, using a methanolic extract (MES), a fraction rich in flavonoids (FRF) and a precipitate from Spondias sp.leaves. The antioxidant activity of them was valued to evaluate their free radical scavenger capacity by DPPH test, whereas MES and FRF were used to evaluate while the preventive action on carbon tetrachloride (CCl4)-induced hepatotoxicity. Seven groups (n=5) of female Wistar rats were used as follows: control group, CCl4-intoxicated group treated with EMS (500 mg/kg) for 7 days, three CCl4-intoxicated groups treated with FRF (25, 50 and 75 mg/kg) for 7 days and the CCl4-intoxicated group treated with Legalon ® (silimarina; (phytotherapeutic reference) (50 mg/kg; 7 days). MES and FRF showed a protective action against liver injury induced by CCl4, being observed a significant reduction of serum enzyme activity marker of liver damage (alanine transaminase and aspartate transaminase). On the other hand, the lipid peroxidation (SRAT) decrease, as well as the increase of glutathione content and enzyme activity of antioxidant defense system (SOD, CAT, GPx) toward near normal values indicated the ability of EMS to restore the oxidative imbalance induced by CCl4. The histological analysis confirmed the hepatoprotection, compared to degenerative changes in CCl4-treated group. This hepatoprotetor effect was similar to that shown by Legalon®. The in vitro high antioxidant capacity of extract (93.16 ± 1.00%) showed analogous results to those obtained by Carduus marianus BHT (reference standard). This fact explains the obtained results in vivo. Although no antimicrobial activity was detected, EMS and FRF promoted the antinociceptive effect induced in the second phase by the intraplantar formalin test, evidencing the anti-inflammatory action; confirmed by the carrageenan-induced peritonitis model. The evaluation of the mechanical allodynia (CFA a 80%) demonstrated the involvement of the Spondias sp. chemical composition in the anti-inflammatory activity toward the acute processes. The acute exposure and repeated dose during 28 days did not produce significant changes in the parameters that evaluate toxicity. Together the experimental results reveal, that Spondias sp. leaf extracts have a promising potential in pharmaceutical area, and due to its non-toxic condition present efficiency and security

DFT study of alpha-alanine as a function of the medium polarity

The zwitterionic (Z) form, neutral (N) form and transition structure (TS) connecting N to Z, have been studied at the B3LYP/6-31++G** level of calculation by using the SCRF methodology. The intramolecular proton transfer from oxygen to nitrogen atoms of alpha -alanine and vibrational spectrum were analyzed in the different environments employed: acetonitrile, ethanol, carbon tetrachloride and gas phase. The Z species is a stationary point in acetonitrile and ethanol, but not in carbon tetrachloride and gas phase media. The geometry of N, Z and TS was similar in acetonitrile and ethanol. The vibrational spectrum of Z was similar in the two solvents studied. (C) 2001 Elsevier B.V. B.V. All lights reserved.

Uso da palha de carnaúba em revestimento de dutos

In this work, the plant species Copernicia prunifera (Miller) H. E. Moore (carnauba), naturally occurring which prevails in the northeast region of Brazil was the subject of studies aiming its use as external coating of pipelines used in petroleum industry. The part of the plant worked were the leaves, also called straw, which were coated with resinous material. For this purpose, it was necessary to evaluate the effectiveness of the use of acrylic resins in the straw carnauba coating. The properties of the untreated carnauba straw and chemically treated with sodium hydroxide, hexane and carbon tetrachloride were investigated by ATRFTIR, SEM and thermal analysis. The first two techniques showed that treatment with solvents has caused major changes in the straw surface, while the thermal analysis indicated that the sodium hydroxide caused variations in thermal stability of straw constituents. Water absorption measurements showed that treatments have accelerated the absorption process and the reduction of contact angle values for treated samples with solvents indicated higher hidrophilicity of straw. The tensile tests showed lower values of elastic modulus and tensile strength for treated samples. Furthermore, coatings using pure commercial resins A and B as well as the formulations with clay were applied in straw and they were examined once again through thermal analysis, water absorption measurements, contact angle and mechanical tests. To analyze the effect of heat ageing, samples were subjected to tensile tests again in order to assess its resistance. The results showed that the resins/clay formulations increased thermal stability of straw, they promoted a good impermeabilization and caused significant decrease in the values of elastic modulus and tensile strength. Evaluating the ageing effect on the mechanical properties, it has been showed good recovery to the coated straw with the formulations A 60 and A 80% in modulus and tensile strength values and elongation at break values have remained very close. It is thus concluded that the carnauba straw can be used as a coating of pipelines with significant cost savings, since there is no need for pretreatment for its use and shows itself as a viable biotechnology alternative, contributing to the quality of coatings material and environment preservation.

Alternative saline solutions to float foraminiferal tests

Flotation has been widely used in studies of recent foraminifera in order to concentrate tests and save time during picking. In this paper, four flotation agents with different densities were compared: (1) trichloroethylene, TCE (C2HCl3), with a density of 1.46 g mL(-1); (2) sodium nitrate/sodium thiosulfate solution, SNT (NaNO3 + Na2O3S2 center dot 5H(2)O), with a density of 1.46 g mL(-1); (3) zinc chloride Solution, ZC (ZHCl(2)), with a density of 1.70 g mL(-1); and (4) sodium polytungstate solution, SPT (3Na(2)WO(4) center dot 9WO(3) center dot 5H(2)O), with a density of 2.50 g mL(-1). Comparison was carried out by means of qualitative and quantitative data. Results showed that ZC and SPT were the best flotation agents, recovering 91% and 96% of the total tests, respectively, whereas TCE and SNT recovered 59.1% and 72.8%, respectively. Both quantitative and qualitative results significantly improved with a higher density of the flotation liquid. Therefore, substitution of TCE with ZC or SPT solutions is strongly encouraged, because they are, additionally, less harmful to health and the environment. ZC is the most cost-effective, since its results were not significantly different from those of the SPT treatment. Carbon tetrachloride (CCl4) was not considered in this comparative study, because it has been banned in many countries and it is highly harmful to health and the environment.

Effect of Brazilian propolis on experimental hepatotoxicity in rats

Propolis (bee glue) is one of the major hive products of bees and is rich in flavonoids, which are known for their antioxidant activities. The aim of this study was to evaluate the hepatoprotective effects of the ethanolic extract of propolis (EEP) against experimental carbon tetrachloride (CCl 4)-induced liver toxicity in rats by means of biochemical indices. The animals were divided into 4 groups: GI= received mineral oil; GII= CCl 4(4mL/kg; Lp., single dose) treated; GIII= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Tetragonisca angustula, daily for 3 days and GIV= CCl4 (4mL/kg; i.p., single dose) treatment followed by ethanolic extract of propolis (100mg/kg) for gavage from the species Nannotrigonea testaceicornes, daily, for 3 days. The serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol and triacylglycerols were estimated after 3 days. CCl 4 caused a maximum increase (p<0,01) above biochemical parameters. As compared to CCl4 group (GII), the EEP (GIII and GIV) showed reduction in cholesterol, triacylglycerol, ALT, AST and alkaline phosphatase activity in the serum. In conclusion, these data indicate that EEP improved the dyslipidaemia, moreover, significantly attenuated increases in serum ALT and AST activities in rats with liver damage induced by carbon tetrachloride.

Tratamento hepatoprotetor favorece a resposta leucocitária de ratos wistar intoxicados por CCL4

The aims of this investigation was to evaluate the effect of hepatoprotective treatments with a compound prepared by the association of N-Acetyl DL-Methionine (5%) + Choline chloride (2%) + Caffeine (1%) + Thiamine hydrochloride (1%) + Nicotinamida (0,5%)+ Pyridoxine hydrochloride (0.04%), administered through intramuscular (IM) route, at doses of 0.2, 0.6 and 1.0 mL/kg of BW, through the study of leukocytes responses in rats submitted to acute intoxication with CCl4. 147 females were randomized into 21 groups, performing five different treatments, which were evaluated seven animals in four periods: two, four, six and eight days after CCl4-induced intoxication. In this study, it was observed absolute eosinophilia and monocytosis in animals untreated and treated with the lowest dose of 0.2 mL. These responses were significantly better in animals treated with 0.6 and 1.0 mL/ kg BW. The untreated animals showed thrombocytopenia, when compared to treated animals. Absolute neutropenia and lymphocytosis was observed in all rats intoxicated with CCl4, there is no difference among treatments. The analysis of white blood cells demonstrated that the hepatoprotective treatments favored the leukocyte response, by act beneficially on the population of these cells, supporting the hypothesis that these events may reduce the deleterious effects in liver tissue after intoxication by CCl4.

Atividade antioxidante da bixina sobre os danos causados pelo tetracloreto de carbono no fígado de ratos

Pós-graduação em Ciência Animal - FMVA

Efeito da suplementação de vitamina D no desenvolvimento de lesões pré-neoplásicas quimicamente induzidas em ratos Wistar macho

Pós-graduação em Biologia Geral e Aplicada - IBB

Prevention of rat liver fibrosis and carcinogenesis by coffee and caffeine

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

Aplicação do método Phitoscreening para determinação de hidrocarbonetos clorados no município de Porto Feliz - SP

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Investigation of I -compounds (newly discovered indigenous DNA modifications) by different approaches

I-compounds are newly discovered covalent DNA modifications detected by the $\sp{32}$P-postlabeling assay. They are age-dependent, tissue-specific and sex-different. The origin(s), chemistry and function(s) of I-compounds are unknown. The total level of I-compounds in 8-10 month old rat liver is 1 adduct in 10$\sp7$ nucleotides, which is not neglectable. It is proposed that I-compounds may play a role in spontaneous tumorigenesis and aging.^ In the present project, I-compounds were investigated by several different approaches. (1) Dietary modulation of I-compounds. (2) Comparison of I-compounds with persistent carcinogen DNA adducts and 5-methylcytosine. (3) Strain differences of I-compounds in relation to organ site spontaneous tumorigenesis. (4) Effects of nongenotoxic hepatocarcinogenes on I-compounds.^ It was demonstrated that the formation of I-compounds is diet-related. Rats fed natural ingredient diet exhibited more complex I-spot patterns and much higher levels than rats fed purified diet. Variation of major nutrients (carbohydrate, protein and fat) in the diet, produced quantitative differences in I-compounds of rat liver and kidney DNAs. Physiological level of vitamin E in the diet reduced intensity of one I-spot compared with vitamin E deficient diet. However, extremely high level of vitamin E in the diet gave extra spot and enhanced the intensities of some I-spots.^ In regenerating rat liver, I-compounds levels were reduced, as carcinogen DNA adducts, but not 5-methylcytosine, i.e. a normal DNA modification.^ Animals with higher incidences of spontaneous tumor or degenerative diseases tended to have a lower level of I-compounds.^ Choline devoid diet induced a drastic reduction of I-compound level in rat liver compared with choline supplemented diet. I-compound levels were reduced after multi-doses of carbon tetrachloride (CCl$\sb4$) exposure in rats and single dose exposure in mice. An inverse relationship was observed between I-compound level and DNA replication rate. CCl$\sb4$-related DNA adduct was detected in mice liver and intensities of some I-spots were enhanced 24 h after a single dose exposure.^ The mechanisms and explanations of these observations will be discussed. I-compounds are potentially useful indicators in carcinogenesis studies. ^

Oil slicks and hydrocarbons in sea water

Surface and upper-layer pollution of seas and oceans by crude oil and refinery products is under study by investigators in many countries. The Intergovernmental Oceanographic Commission (IOC) and World Meteorological Organization (WMO) have prepared an international experimental project that is to be carried out within the framework of the Integrated Global Oceanic Station System (IGOSS). The purpose of the project is to prepare a picture of distribution and dynamics of oil pollution. Parameters to be observed include: oil patches (slicks), floating lumps of tar on the surface, and hydrocarbons emulsified and dissolved in water. Cruise 22 of R/V Akademik Kurchatov took the ship through regions being the most suitable for pollution studies. They were conducted from March through June 1976. On the cruise, oil slicks were observed visually by a procedure recommended by the international program. Areas of the slicks were determined from speed of the ship and time required to cross them. Surface samples were taken along the path of the ship for determination of concentrations of dissolved and emulsified hydrocarbons in water. In addition, samples were taken from deep water by a 7-liter vinyl water bottle at 17 stations. Hydrocarbons present in the samples were extracted immediately with carbon tetrachloride. Final determination of hydrocarbons was made by infrared spectrophotometry. This method is currently accepted in the Soviet Union in an arbitration capacity for determination of petroleum products dissolved and emulsified in sea water. Infrared spectrophotometry is used to determine hydrocarbons containing methyl and methylene groups, but they are not identified as to origin.

Strain-specific differences in mouse hepatic wound healing are mediated by divergent T helper cytokine responses

Hepatic fibrosis represents the generalized response of the liver to injury and is characterized by excessive deposition of extracellular matrix. The cellular basis of this process is complex and involves interplay of many factors, of which cytokines are prominent. We have identified divergent fibrosing responses to injury among mouse strains and taken advantage of these differences to examine and contrast T helper (Th)-derived cytokines during fibrogenesis. Liver injury was induced with carbon tetrachloride, fibrosis was quantitated, and Th1/Th2 cytokine mRNAs measured. Liver injury in BALB/c mice resulted in severe fibrosis, whereas C57BL/6 mice developed comparatively minimal fibrosis. Fibrogenesis was significantly modified in T and B cell-deficient BALB/c and C57BL/6 severe combined immunodeficient (SCID) mice compared with wild-type counterparts, suggesting a role of Th subsets. Fibrogenic BALB/c mice exhibited a Th2 response during the wounding response, whereas C57BL/6 mice displayed a Th1 response, suggesting that hepatic fibrosis is influenced by different T helper subsets. Moreover, mice lacking interferon γ, which default to the Th2 cytokine pathway, exhibited more pronounced fibrotic lesions than did wild-type animals. Finally, shifting of the Th2 response toward a Th1 response by treatment with neutralizing anti-interleukin 4 or with interferon γ itself ameliorated fibrosis in BALB/c mice. These data support a role for immune modulation of hepatic fibrosis and suggest that Th cytokine subsets can modulate the fibrotic response to injury.

Plasminogen deficiency leads to impaired remodeling after a toxic injury to the liver

Cellular proliferation and tissue remodeling are central to the regenerative response after a toxic injury to the liver. To explore the role of plasminogen in hepatic tissue remodeling and regeneration, we used carbon tetrachloride to induce an acute liver injury in plasminogen-deficient (Plgo) mice and nontransgenic littermates (Plg+). On day 2 after CCl4, livers of Plg+ and Plgo mice had a similar diseased pale/lacy appearance, followed by restoration of normal appearance in Plg+ livers by day 7. In contrast, Plgo livers remained diseased for as long as 2.5 months, with a diffuse pale/lacy appearance and persistent damage to centrilobular hepatocytes. The persistent centrilobular lesions were not a consequence of impaired proliferative response in Plgo mice. Notably, fibrin deposition was a prominent feature in diseased centrilobular areas in Plgo livers for at least 30 days after injury. Nonetheless, the genetically superimposed loss of the Aα fibrinogen chain (Plgo/Fibo mice) did not correct the abnormal phenotype. These data show that plasminogen deficiency impedes the clearance of necrotic tissue from a diseased hepatic microenvironment and the subsequent reconstitution of normal liver architecture in a fashion that is unrelated to circulating fibrinogen.