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The paper explores cognitive neuroscience contributions to the education of adult and young people. Advances in brain understanding make new contributions in the educational field, the will of exploring these supports this text based on the analysis of a participatory research process developed with young and adult people teachers. The study addresses outstanding topics in neuroscience such as learning environment, memory and remembering, motivation and emotions, anxiety and stress, attention system and space organization as they relate to the diversity and complexity the education of adult and young people faces nowadays. The text concludes that advancing in brain understanding requires an interdisciplinary vision which articulates biological sciences with advances from sociology, psychology, and anthropology incorporating as a central perspective the educators’teaching reflection., El artículo indaga en los aportes de la neurociencia cognitiva a la educación de personas jóvenes y adultas. Los avances en la comprensión del cerebro abren contribuciones nuevas en el ámbito educativo, el deseo de explorar en estas contribuciones sustenta este texto basado en el análisis de un proceso de investigación participativa desarrollado con docentes de educación de personas jóvenes y adultas. El estudio aborda temas destacados en neurociencia como son ambiente de aprendizaje, memoria y recuerdo, motivación y emociones, ansiedad y estrés, sistema atencional y organización del espacio relacionándolo con la diversidad y complejidad que enfrenta la educación de personas jóvenes y adultas. El texto concluye que para avanzar en la comprensión del cerebro se requiere una visión interdisciplinaria que articule las ciencias biológicas con los avances desde la sociología, psicología y antropología incorporando como perspectiva central la reflexión pedagógica de las y los educadores.

Nitrofurantoin (NFT), an antimicrobial drug used for the treatment of uncomplicated lower urinary tract infections, produces gastrointestinal adverse effects that compromise treatment compliance. These adverse effects are related to oxidative stress generated by the enzymatic reduction of NFT.

A simple and fast spectrophotometric methodology able to quantify superoxide released by NADPH oxidase from differentiated promyelocytic leukaemia (HL-60) cells using pyrogallol red is described.The latter is based on the known stoichiometry of the reaction between superoxide and pyrogallol red and the inability of pyrogallol red to react with hydrogen peroxide. In addition, we developed a 96-wells microplate-based method able to determine NADPH oxidase activity. Using this method, we determined pharmacological properties of the NADPH oxidase inhibitors VAS2870 and diphenyleneiodonium and the obtained IC50 values were in good agreement with previous reported data. NOX2 is highly expressed in differentiated promyelocytic leukaemia cells, whereas other isoforms are not detected or expressed at low amounts. Likewise, this methodology may be a useful assay for NOX2 inhibitor screening. NADPH oxidases are involved in several physiological and pathological processes, rendering its pharmacological modulation an attractive research target. In this context, this simple assay can be used for NADPH oxidase inhibitor screening as well as aiding in the study of different biological conditions that involve NADPH oxidase activity.

Adverse reactions of acetaminophen have been associated to oxidative stress, which may be elicited by reactive oxygen species (ROS) and/or production of the metabolite NAPQI. Both phenomena would arise through the activity of liver cytochrome P450 (CYP450) system, but their contribution to this oxidative stress is yet to be clarified. A NADPH oxidase activity has been proposed in rat liver microsomes. This activity may be due to the presence of NAD(P)H oxidase (NOX) isoforms in liver endoplasmic reticulum. Both NOX and the CYP450 system activities can catalyze ROS generation using NADPH as a cofactor. Therefore, acetaminophen biotransformation, which requires NADPH, may promote ROS generation through either activity or both. To discriminate between these possibilities, rat liver microsomes were incubated with acetaminophen and NADPH in the presence or absence of specific inhibitors. Incubation with NADPH and acetaminophen elicited lipid peroxidation and decreased thiol content and glutathione-S-transferase (GST) activity. The NOX inhibitors apocynin and plumbagin prevented all these phenomena but the decrease in thiol content. In contrast, this decrease was completely prevented by the specific CYP450 system inhibitor SKF-525A. These data suggest that ROS generation following incubation of microsomes with acetaminophen and NADPH appears to be mainly caused by a NOX activity. In light of these data, toxicity of acetaminophen is discussed.

Chenopodium quinoa Willd (quinoa) has been a source of food for millennia by the Andes region native population. Because of its bitter taste, quinoa seeds are commercialized without their coat for human consumption. Quinoa coats are surfactant sub-products of the quinoa food industry, which have been only characterized to contain triterpene saponins. We postulated that this coat should also contain antioxidant molecules as part of the defense system of the quinoa seed. We found that a quinoa seed coats hydroalcoholic extract, displayed thiol compounds in addition to polyphenols, recognized antioxidants. Accordingly, it inhibited microsomal lipid peroxidation and the loss of microsomal thiol content, both oxidative phenomena promoted by Cu 2+ /ascorbate. Microsomal glutathione S -transferase (GST) is inhibited by reducing agents, which decrease the content of catalytically active disulfide-linked dimers. The effects of this quinoa extract on microsomal GST are consistent with it displaying disulfide reducing properties. The occurrence of thiol compounds in this quinoa extract is discussed in terms of the potential of their antioxidant properties.

Iron and copper ions, in their unbound form, may lead to the generation of reactive oxygen species via Haber-Weiss and/or Fenton reactions. In addition, it has been shown that copper ions can irreversibly and non-specifically bind to thiol groups in proteins. This non-specific binding property has not been fully addressed for iron ions. Thus, the present study compares both the pro-oxidant and the non-specific binding properties of Fe(3+) and Cu(2+), using rat liver cytosol and microsomes as biological systems. Our data show that, in the absence of proteins, Cu(2+)/ascorbate elicited more oxygen consumption than Fe(3+)/ascorbate under identical conditions. Presence of cytosolic and microsomal protein, however, differentially altered oxygen consumption patterns. In addition, Cu(2+)/ascorbate increased microsomal lipid peroxidation and decreased cytosolic and microsomal content of thiol groups more efficiently than Fe(3+)/ascorbate. Finally, Fe(3+)/ascorbate and Cu(2+)/ascorbate inhibited in different ways cytosolic and microsomal glutathione S-transferase (GST) activities, which are differentially sensitive to oxidants. Moreover, in the absence of ascorbate, only Cu(2+) decreased the content of cytosolic and microsomal thiol groups and inhibited cytosolic and microsomal GST activities. Catechin partially prevented the damage to thiol groups elicited by Fe(3+)/ascorbate and Cu(2+)/ascorbate but not by Cu(2+) alone. N-Acetylcysteine completely prevented the damage elicited by Cu(2+)/ascorbate, Fe(3+)/ascorbate and Cu(2+) alone. N-Acetylcysteine also completely reversed the damage to thiol groups elicited by Fe(3+)/ascorbate, partially reversed that of Cu(2+)/ascorbate but failed to reverse the damage promoted by Cu(2+) alone. Our data are discussed in terms to the potential damage that the accumulation of iron and copper ions can promote in biological systems.

Objective—To evaluate the antioxidant effects of synovial fluid (SF) pooled from metacarpophalangeal joints of healthy horses or horses with various pathological conditions, and to compare them with the antioxidant effects of hyaluronic acid (HA) and chondroitin sulfate (CS). Sample Population—SF from 1 metacarpophalangeal joint was obtained from 42 horses immediately after humane slaughter. Samples were classified into 3 groups on the basis of origin: healthy joints or joints with chronically damaged cartilage or vascularly congested synovial membranes as detected via macroscopic evaluation. Procedures—Antioxidant effects were evaluated by use of rat liver microsomal fractions treated with Fe3+-ascorbate as a free radical generator system leading to oxidative stress. Amounts of thiobarbituric-reactive substances and glutathione transferase (GSH-T) conjugation activity were measured. Results—SF from healthy and chronically damaged joints inhibited microsomal lipid peroxidation, whereas SF from joints with congested synovial membranes had only a slight effect. Hyaluronic acid and CS did not inhibit microsomal lipid peroxidation. Moreover, GSH-T activity was detected in all SF samples, which had similar activity regardless of disease status. All SF samples as well as HA and CS protected rat microsomal GSH-T activity against oxidative damage. Only SF samples from joints with congested synovial membranes protected microsomal thiols against oxidation, an effect also evident with HA and CS. Conclusions and Clinical Relevance—The antioxidant mechanisms associated with the response to metacarpophalangeal joint damage in horses appeared to act on different targets, depending on whether the damage was acute or chronic.

1,4-Dihydropyridines (DHPs) used in the treatment of cardiovascular diseases, are calcium channel antagonists and also antioxidant agents. These drugs are metabolized through cytochrome P(450) oxidative system, majority localized in the hepatic endoplasmic reticulum. Several lipophilic drugs generate oxidative stress to be metabolized by this cellular system. Thus, DHP antioxidant properties may prevent the oxidative stress associated with hepatic biotransformation of drugs. In this work, we tested the antioxidant capacity of several synthetic nitro-phenyl-DHPs. These compounds (I-IV) inhibited the microsomal lipid peroxidation, UDPGT oxidative activation and microsomal thiols oxidation; all phenomena induced by Fe(3+)/ascorbate, a generator system of oxygen free radicals. As the same manner, these compounds inhibited the oxygen consumption induced by Cu(2+)/ascorbate in the absence of microsomes. Furthermore, compound III (2,6-dimethyl-4-(4-nitrophenyl)-1,4-dihydropyridin-3,5-ethyl-dicarboxylate) and compound V (N-ethyl-2,6-dimethyl-4-(4-nitrophenyl)-1,4-dihydropyridin-3,5-methyl-dicarboxylate) inhibited the microsomal lipid peroxidation induced by Nitrofurantoin and naphthalene in the presence of NADPH. Oxidative stress induced on endoplasmic reticulum may alter the biotransformation of drugs, so, modifying their plasmatic concentrations and therapeutic effects. When drugs which are activated by biotransformation are administered together with antioxidant drugs, such as DHPs, oxidative stress induced in situ may be prevented.

H 2 O 2 inactivation of particular GST isoforms has been reported, with no information regarding the overall effect of other ROS on cytosolic GST activity. The present work describes the inactivation of total cytosolic GST activity from liver rats by the oxygen radical-generating system Cu 2+ /ascorbate. We have previously shown that this system may change some enzymatic activities of thiol proteins through two mechanisms: ROS-induced oxidation and non-specific Cu 2+ binding to protein thiol groups. In the present study, we show that nanomolar Cu 2+ in the absence of ascorbate did not modify total cytosolic GST activity; the same concentrations of Cu 2+ in the presence of ascorbate, however, inhibited this activity. Micromolar Cu 2+ in either the absence or presence of ascorbate inhibited cytosolic GST activity. Kinetic studies show that GSH but no 1-chloro-2,4-dinitrobenzene prevent the inhibition on cytosolic GST induced by micromolar Cu 2+ either in the absence or presence of ascorbate. On the other hand, NEM and mersalyl acid, both thiol-alkylating agents, inhibited GST activity with differential reactivity in a dose-dependent manner. Taken together, these results suggest that an inhibitory Cu 2+ -binding effect is likely to be negligible on the overall inhibition of cytosolic GST activity observed by the Cu 2+ /ascorbate system. We discuss how modification of GST-thiol groups is related to the inhibition of cytosolic GST activity.

Toxic effects of several nitro-aryl drugs are attributed to the nitro-reduction that may be suffered in vivo, a reaction that may be catalysed by different reductases. One of these enzymes is NADPH–cytochrome P450 reductase, which belongs to the cytochrome P450 oxidative system mainly localized in the endoplasmic reticulum of the hepatic cell. This system is responsible for the biotransformation of oxidative lipophilic compounds, so that oxidative and reductive metabolic pathways of lipophilic nitro-aryl drugs can take place simultaneously. Because of the affinity of nitro-aryl drugs (xenobiotics) for the endoplasmic reticulum, we propose this subcellular organelle as a good biological system for investigating the toxicity induced by the biotransformation of these or another compounds. In this work we used rat liver microsomes to assess the oxidative stress induced by nitro-aryl drug biotransformation. Incubation of microsomes of rat liver with nifurtimox and nitrofurantoin in the presence of NADPH induced lipoperoxidation, UDP-glucuronyltransferase activation and an increase in the basal microsomal oxygen consumption. Nitro-aryl-1,4-dihydropyridines did not elicit these prooxidant effects; furthermore, they inhibited lipoperoxidation and oxygen consumption induced by Fe3+/ascorbate. Nifurtimox and nitrofurantoin modified the maximum absorption of cytochrome P450 oxidase and inhibited p-nitroanisole O-demethylation, an oxidative reaction catalysed by the cytochrome P450 system, signifying that oxidation may proceed in a similar way to that described for nitro-aryl-1,4-dihydropyridines. Thus the balance between lipophilic nitro-aryl drug oxidation and reduction may be involved in the potential oxidative stress induced by biotransformation. Copyright © 2004 John Wiley & Sons, Ltd.

Metallothionein (MT) and reduced glutathione (GSH) play an important role in the intracellular handling of copper by preventing the generation and favouring the removal of copper-derived free radicals. The present study addressed the changes in MT and GSH that follow chronic (2 or 5 weeks) exposure of human hepatoblastoma cells (HepG2) to excess copper. Copper treatment (100 microM, 2 weeks) led to a 28-fold elevation in intracellular copper. Concomitantly, cells exhibited a seven-fold increase in total MT and an increment in its saturation with copper from 45 to 86%. Around 38% of copper in the cytosolic fraction could be accounted for by MT. GSH equivalents were substantially lowered (to 37% of basal levels) in treated cells, with only part of it being accounted for by an increase in GSSG. Copper-treatment induced no changes in catalase or GSH-peroxidase activities but it was associated with a small reduction in SOD (20%) and GSH-reductase (26%) activities. Copper-loaded cells did not differ from controls in their basal oxidative tone; however, when exposed to tert-butylhydroperoxide they exhibited a markedly greater susceptibility to undergo both oxidative stress and cell lysis. It is proposed that chronic exposure of HepG2 cells to excess copper is accompanied by "adaptive changes" in GSH and MT metabolism that would render cells substantially more susceptibility to undergo oxidative stress-related cytotoxicity.

Tumor cells principally exhibit increased mitochondrial transmembrane potential (ΔΨ(m)) and altered metabolic pathways. The therapeutic targeting and delivery of anticancer drugs to the mitochondria might improve treatment efficacy. Gallic acid exhibits a variety of biological activities, and its ester derivatives can induce mitochondrial dysfunction. Four alkyl gallate triphenylphosphonium lipophilic cations were synthesized, each differing in the size of the linker chain at the cationic moiety. These derivatives were selectively cytotoxic toward tumor cells. The better compound (TPP(+)C10) contained 10 carbon atoms within the linker chain and exhibited an IC50 value of approximately 0.4-1.6 μM for tumor cells and a selectivity index of approximately 17-fold for tumor compared with normal cells. Consequently, its antiproliferative effect was also assessed in vivo. The oxygen consumption rate and NAD(P)H oxidation levels increased in the tumor cell lines (uncoupling effect), resulting in a ΔΨ(m) decrease and a consequent decrease in intracellular ATP levels. Moreover, TPP(+)C10 significantly inhibited the growth of TA3/Ha tumors in mice. According to these results, the antineoplastic activity and safety of TPP(+)C10 warrant further comprehensive evaluation.

Nifurtimox is a drug used against Chagas' disease. Its biotransformation in intact epimastigotes, microsomal membranes and cytosolic fractions of Trypanosoma cruzi Tulahuen strain was examined. Nifurtimox reduction by intact cells occurred at a rate of 23.19 nmol mg protein−1 h−1. The nitroreductase activity was found to be distributed among the microsomal and cytosolic fractions, being mostly NADPH dependent. Also, in the present paper intact cells and membrane-enriched fraction of T. cruzi were used to prove radical species formation. Results from these experiments show that the increase of the NADPH-oxygen consumption induced by the drug, was not dependent on the respiratory chain of the parasite. On the other hand, the nifurtimox-induced H2O2 production was significant, providing an indirect proof of the nitro anion formation.

Nitrofurantoin is used in the antibacterial therapy of the urinary tract. This therapy is associated with various adverse effects whose mechanisms remain unclear. Diverse studies show that the nitro reductive metabolism of nitrofurantoin leads to ROS generation. This reaction can be catalyzed by several reductases, including the cytochrome P450 (CYP450) reductase. Oxidative stress arising from this nitro reductive metabolism has been proposed as the mechanism underlying the adverse effects associated with nitrofurantoin. There is, however, an apparent paradox between these findings and the ability of nitrofurantoin to inhibit lipid peroxidation provoked by NADPH in rat liver microsomes. This work was aimed to show the potential contribution of different enzymatic systems to the metabolism of this drug in rat liver microsomes. Our results show that microsomal lipid peroxidation promoted by NADPH is inhibited by nitrofurantoin in a concentration-dependent manner. This suggests that the consumption of NADPH in microsomes can be competitively promoted by lipid peroxidation and nitrofurantoin metabolism. The incubation of microsomes with NADPH and nitrofurantoin generated 1-aminohidantoin. In addition, the biotransformation of a classical substrate of CYP450 oxidative system was competitively inhibited by nitrofurantoin. These results suggest that nitrofurantoin is metabolized through CYP450 system. Data are discussed in terms of the in vitro redox metabolism of nitrofurantoin.

The electrochemical properties of nifuroxazide have been investigated in aqueous and aqueous-DMF mixed solvents. In aqueous media, a single, irreversible four-electron reduction occurs to give the hydroxylamine derivative. In mixed media, a reversible one-electron reduction to form a nitro radical anion takes place. Cyclic voltammetric studies show that the anion radical product is stable, although the nitro radical anion intermediate shows a tendency to undergo further chemical reactions. A comparison with the voltammetric behaviour of other nitrofurans such as nifurtimox, nitrofurazone and furazolidone is made. The electrochemically-obtained parameters are correlated with the in vivo studies of oxygen consumption on Trypanosoma cruzi cell suspensions.

BHA (2(3)-tert-butyl-4-hydroxyanisole) produced inhibition of both culture growth and oxygen consumption, NAD(P) reduction and cytochrome b oxidation, on intact epimastigotes of Trypanosoma cruzi. BHA inhibited respiration and reduced NAD(P) in intact T. cruzi trypomastigotes. Oxidative phosphorylation of in situ mitochondria of epimastigotes was inhibited by BHA and this effect was liberated by the addition of ascorbate + TMPD. The incorporation of rhodamine-123 to mitochondria of living epimastigotes was diminished by BHA. These results suggest that the basis of the trypanocidal effects of BHA could be due to the blockage of the mitochondrial electron transport chain on the segment previous to cytochrome c. We postulate that the mechanism of action of BHA could be by mimicking coenzyme-Q (ubiquinone).

Se determino la presencia de isoformas de sintasa del oxido nitrico (NOS) en membrana sinovial de articulacion metacarpofalangica equina. Se considero un grupo normal NN (n=13) sin dano observable en las estructuras articulares; grupo NA (n = 8) con la membrana sinovial congestiva; grupo AN (n=10) con el cartilago danado y membrana sinovial normal; AA (n = 8) con el cartilago y membrana sinovial alterados. La deteccion de las isoformas de NOS se realizo mediante “Western Blot” utilizando anticuerpos monoclonales. La quimioluminiscencia generada fue medida y expresada en unidades densitometricas relativas (UDR) que corresponde a la razon entre los pixeles de cada muestra y de un patron interno. La NOS I de 145 kDa se encontro en todas las muestras analizadas. La NOS III de 140 kDa se detecto solo en un porcentaje de las muestras (NN = 77%; NA = 63%; AN = 30%; AA = 100%) y no se encontro una diferencia significativa entre los grupos. No se detecto la NOS II en ninguna de las muestras analizadas. Se concluye que la NOS I y NOS III son constitutivas de la membrana sinovial, mientras que la NOS II no se detecta, posiblemente debido a que no existe una inflamacion aguda en los grupos analizados.

The bleaching of the pyrogallol red (PGR) dye mediated by superoxide anion radicals ( O 2 - ) generated from the xanthine/xanthine oxidase system (X/XO) was studied by UV-visible spectrophotometry. The absorption band (at 540 nm) of PGR quickly decreased in the presence of X/XO, implying an efficient reaction of O 2 - with PGR. The process was unaffected by catalase (CAT), but completely abolished by superoxide dismutase (SOD). A mechanism of the reaction involving the consumption of one PGR molecule by two O 2 - to generate one molecule of H2O2 is proposed. PGR was used as a probe to estimate the rate of O 2 - generation in redox cycling reactions of a series of nitro compounds mediated by rat liver microsomes. The consumption of PGR induced by the redox cycling of nitrofurantoin was totally eliminated by the addition of SOD but unaffected by CAT. The initial rate of consumption of PGR mediated by the redox cycling of others nitro derivatives follows the order: furazolidindione > nitrofurantoin > nifurtimox > benznidazole > chloramphenicol. We concluded that PGR can be used as a probe to estimate the release of O 2 - from enzymatic systems or from the redox cycling of nitro compounds.

Copper toxicity has been associated to the capacity of free copper ions to catalyze the production of superoxide anion and hydroxyl radical, reactive species that modify the structure and/or function of biomolecules. In addition, nonspecific Cu2+-binding to thiol enzymes, which modifies their catalytic activities, has been reported. Cytochrome P450 (CYP450) monooxygenase is a thiol protein that binds substrates in the first and limiting step of CYP450 system catalytic cycle, necessary for the metabolism of lipophilic xenobiotics. Therefore, copper ions have the potential to oxidize and bind to cysteinyl residues of this monooxygenase, altering the CYP450 system activity. To test this postulate, we studied the effect of Cu2+ alone and Cu2+/ascorbate in rat liver microsomes, to independently evaluate its nonspecific binding and its pro-oxidant effects, respectively. We assessed these effects on the absorbance spectrum of the monooxygenase, as a measure of structural damage, and p-nitroanisole O-demethylating activity of CYP450 system, as a marker of functional impairment. Data obtained indicate that Cu2+ could both oxidize and bind to some amino acid residues of the CYP450 monooxygenase but not to its heme group. The differences observed between the effects of Cu2+ and Cu2+/ascorbate show that both mechanisms are involved in the catalytic activity inhibition of CYP450 system by copper ions. The significance of these findings on the pharmacokinetics and pharmacodynamics of drugs is discussed. Copyright © 2009 John Wiley & Sons, Ltd.

Glutathione S-transferases (GSTs) are isoenzymes occurring in the cytoplasm and as integral membrane proteins. In addition to their role in drug metabolism by conjugating electrophilic and lipophilic compounds with glutathione (GSH), these enzymes display multiple functions in cells, including antioxidant action. It has been generalized that reactive oxygen species (ROS) inhibit cytosolic GSTs and activate microsomal GSTs; some evidence shows, however, that different ROS-generating systems can inhibit microsomal GST activity. We therefore tested the effect of Fe3+/ascorbate, another ROS-generating system, on cytosolic and microsomal GST activities from rat liver cytosol and microsomes, respectively, and compared it to that of hydrogen peroxide (H2O2). We found that, while both agents displayed similar inhibitory effects on cytosolic GST activity, they promoted opposite effects on microsomal GST activity. Using specific antioxidant enzymes, we corroborated that the effect of Fe3+/ascorbate involves generation of \( {{\text{O}}_2}^{ \bullet - } \) without dismutation into H2O2. Since these ROS have physicochemical properties and redox potentials that are very distinct, their reactivity is different, and their oxidative action is likely to have different targets. We discuss how these properties are related with the oxidative potency of ROS, especially those of \( {{\text{O}}_2}^{ \bullet - } \) and H2O2.

The following phenolic antioxidant food additives were evaluated against Trypanosoma cruzi epimastigotes: BHT, BHA, gallic acid and its methyl, propyl, octyl, and lauryl esters, 2,4-di-tert-butyl-6-(4-methoxybenzyl)-phenol, 4,4′-isopropilidenediphenol, and protocatechuic acid and its ethyl ester. The inhibition of the respiration; the changes in motility, shape, and lysis of the parasites; and the human blood hemolysis caused by these chemicals were studied. Human blood samples experimentally contaminated with 2000 or 150,000 trypomastigotes per milliliter were freed of parasites after treatment for 24 hr at 4 °C with 5 or 10 mM BHT (2,6-di-tert-butyl-4-hydroxytoluene), respectively. Consequently, BHT and other phenolic compounds deserve further study to determine their role in preventing the transmission of Chagas' disease by blood transfusion.

Numerous investigations exist about the alterations that oxygen free radicals can provoke on biomolecules; these modifications can be prevented and/or reversed by different antioxidants agents. On the other hand, 2,2-diphenyl-1-picrylhydrazyl radical (DPPH), a stable nitrogen synthetic radical, is used to evaluate the antioxidant capacity of medicinal herbal products; however, the structural changes that this radical provoke on the herbal active principles are not clear yet. In this work, we compared the redox reactivity of oxygen free radicals and DPPH radical on phospholipids and protein thiol groups present in rat liver microsomes. Cu2+/ascorbate was used as generator system of oxygen free radical and as antioxidant, an extract of Buddleja globosa's leaves. Cu2+/ascorbate provoked microsomal lipid peroxidation, microsomal thiols oxidation and oxygen consumption; all of these phenomena were inhibited by B. globosa extract. On the other hand, DPPH was bleached in different extension by the herbal extract and phosphatidyl choline; beside, DPPH decreased microsomal thiols content, but this phenomenon were not prevented by the herbal extract. Furthermore, DPPH did not induce oxygen consumption and neither modified the oxygen consumption induced by Cu2+/ascorbate. Distinct redox mechanisms may explain the differences between the reactivity of DPPH and oxygen free radicals on biomolecules, which is discussed.

Treatment of hepatic microsomes with Fe(3+)/ascorbate activates UDP-glucuronyltransferase (UGT), a phenomenon totally prevented and reversed by reducing agents. At microM concentrations, iron and copper ions catalyze the formation of ROS through Fenton and/or Haber-Weiss reactions. Unlike iron ions, indiscriminate binding of copper ions to thiol groups of proteins different from the specialized copper-binding proteins may occur. Thus, we hypothesize that incubation of hepatic microsomes with the Cu(2+)/ascorbate system will lead to both UGT oxidative activation and Cu(2+)-binding induced inhibition, simultaneously. We studied the effects of Cu(2+) alone and in the presence of ascorbate on rat liver microsomal UGT activity. Our results show that the effects of both copper alone and in the presence of ascorbate were copper ion concentration- and incubation time-dependent. At very low Cu(2+) (25nM), this ion did not modify UGT activity. In the presence of ascorbate, however, UGT activity was increased. At higher copper concentrations (10 and 50microM), this ion led to UGT activity inhibition. In the presence of ascorbate, 10microM Cu(2+) activated UGT at short incubation periods but inhibited this enzyme at longer incubation times; 50microM Cu(2+) only inhibited UGT activity. Thiol reducing agent 2,4-dithiothreitol prevented and reversed UGT activation while EDTA prevented both, UGT activation and inhibition. Our results are consistent with a model in which Cu(2+)-induced oxidation of UGT leads to the activation of the enzyme, while Cu(2+)-binding leads to its inhibition. We discuss physiological and pathological implications of these findings.

Leaf extracts of Ugni molinae Turcz. are used in the Chilean cosmetic industry on the assumption that they have decongestant, regenerative, and anti-aging properties. A bioassay-guided fractionation of this plant material showed that some extracts have potent anti-inflammatory activities. Further fractionation led to the isolation and identification of betulinic acid, a mixture of ursolic and oleanolic acids, and the 2alpha-hydroxy derivatives alphitolic, asiatic, and corosolic acids. The latter three were evaluated in vivo in the mouse ear assay for their topical anti-inflammatory activity, inducing inflammation with either arachidonic acid (AA) or 12-O-tetradecanoylphorbol-13 acetate (TPA). Only corosolic acid was active in the AA assay, with similar potency to nimesulide, but all three triterpene acids inhibited TPA-induced inflammation with potencies comparable to that of indomethacin.

1. Lipid peroxidation in rat brain slices was induced by Fe+3/ascorbate. 2. Brain lipid peroxidation, as measured by malondialdehyde formation, was inhibited by all the tested nitro aryl 1,4-dihydropyridine derivatives over a wide range of concentrations. The time-course antioxidant effects of the most representative agents were assessed. On the basis of both time-course and IC50 experiments the tentative order of antioxidant activity on rat brain slices could be: nicardipine>nisoldipine> (R,S/S,R)-furnidipine > (R,R/S,S)-furnidipine>nitrendipine>nimodipine> nifedipine. 3. 1,4-Dihydropyridine derivatives that lack of a nitro group in the molecule (isradipine, amlodipine) also inhibited lipid peroxidation in rat brain slices but at higher concentrations than that of nitro-substituted derivatives. 4. All the tested nitroso aryl derivatives [2,6-dimethyl-4-(2-nitrosophenyl)-3,5-pyridinedicar. boxylic acid dimethyl ester (NTP), nitrosotoluene, nitrosobenzene] were more potent inhibitors of lipid peroxidation than were the parent nitro compounds. In conclusion, on the basis of the IC50 values determined, the rank order of antioxidant potency for these derivatives can be established as: ortho-nitrosotoluene>NTP>nitrosobenzene.

1. Isradipine and lacidipine, two new drugs that are members of the nitro-aryl-1,4-dihydropyridine family, produced inhibition of both growth cultures and oxygen consumption on epimastigotes of Trypanosoma cruzi Tulahuen strain, at micromolar concentrations. 2. Isradipine was found to be the most potent derivative in both, in growth cultures (I50 = 20.8 microM) and in vivo oxygen uptake (I50 = 31.1 microM). 3. Diltiazem and verapamil, two well-known calcium channel antagonists, lacked inhibitory activity, even at a 100 microM concentration. 4. The present findings indicate that the trypanocide effects exerted by isradipine and lacidipine are not related with a disruption of the calcium homeostasis of the parasite.

A series of nitro aryl 1,4-dihydropyridine derivatives produced inhibition of both cell growth and oxygen consumption on Tulahuen and LQ strains, and clone Dm 28c of epimastigotes of Trypanosoma cruzi. Nicardipine was found to be the most potent derivative in both growth cell (I50 = 70 microM) and oxygen uptake (I50 = 26 microM in intact parasites, I50 = 325 microM in situ mitochondria). A correlation between the inhibitory effects on the growth cell and the apparent first order kinetic for the uptake of the 1,4-dihypyridine derivatives by T. cruzi epimastigotes was found. Thus, nicardipine, the most potent derivative, exhibited the highest apparent rate constant, ku, (0.043 min-1). On the other hand, no susceptibility differences by strains and clone Dm 28c to the action of these drugs were found.

1. 1. Carboxylesterase activity in Trypanosoma cruzi was found mainly in the microsomal (40%) and the cytosolic fraction (26%). 2. 2. The Vmax for p-nitrophenyl acetate was 28.50 and 17.60 nmol per min and mg of protein for the microsomal and the cytosolic fractions, respectively. 3. 3. The Km was 0.78 mM for the microsomal activity and 0.55 mM for the cytosolic activity. 4. 4. The inhibition rate constant with N-ethylmaleimide were 38.10 M−1 min−1 and 2.56 M−1 min−1 for the cytosolic and the microsomal enzymes, respectively. 5. 5. The rate constants with Paraoxon were 8,360M−1 × min−1 and 32,600 M−1 × min−1. 6. 6. Polyacrylamide gel electrophoresis under nondenaturing conditions showed three bands of microsomal activity with M.W. of 63, 136 and 153 kDA. 7. 7. Similarly, three bands of cytosolic activity with M.W. of 126, 294 and 479 kDA were identified.

1. 1. Comparative studies of the actions of morphine in different mammals have shown that Octodon degus presents an unusual tolerance to this compound. 2. 2. Morphine glucuronidation and N-demethylation in microsomal fractions of Octodon degus were 10.9 and 50.0 nmol of product formed/min/g of wet liver, respectively. In Wistar rat these activities were 10.5 and 12.5, respectively. 3. 3. Microsomal protein and cytochrome P -450 contents were two and four times higher in Octodon degus than in the Wistar rat, respectively. 4. 4. These results may explain the high tolerance to morphine presented by Octodon degus .

1. 1. Microsomes prepared from Triatoma infestans fifth-instar larvae metabolize DDT to a compound similar to Kelthane (metabolite No. 3), and to a derivative of apparently a phenolic nature (metabolite No. 2). 2. 2. The production of both metabolites requires either NADPH or NADH, but only the formation of metabolite No. 3 is nicotinamide—and magnesium— dependent. 3. 3. Optimum pH for the formation of metabolite No. 3 is 8·5, while it is 9·0 for metabolite No. 2. The K m values for each activity were also slightly different. 4. 4. The activity for each metabolite was increased by pretreatment with phenobarbital, but the induction kinetics was different in each case. 5. 5. These observations suggest that two rather than one microsomal enzyme are involved in DDT metabolism in T. infestans .

t-Butyl-4-hydroxyanisole, an antioxidant food additive, inhibited the growth of Trypanosoma cruzi by almost 100% at 0.5 mM concentration. This compound inhibited 70% of oxygen consumption of epimastigotes. The redox level of NAD(P) was shifted to a more reduced state and inversely the redox level ofcytochrome b changed to a more oxidized state. This hydroxyanisole thus is a new electron transport chain inhibitor. This compound and related ones, or the respiratory chain of T. cruzi, may be important in the design of antichagasic drugs.

1. Glutathione (G-SH) concentration, gamma-glutamyltranspeptidase and glutathione S-transferase activities were studied in several strains of T. cruzi epimastigotes. GSH varied from 1.04 mM for the LQ strain to 0.61 mM for the Tulahuen strain. 2. Cultures of the LQ strain presented more resistance to drugs than those of the Tulahuen. It was necessary a concentration of nifurtimox 4 times higher and one of benznidazole 10 times higher in order to inhibit approximately to 50% the growth of LQ strain cultures when compared with the Tulahuen strain. 3. Buthionine sulfoximine decreased the concentration of glutathione to about 50% in the LQ and Tulahuen strains and potentiated the toxicity of nifurtimox and benznidazole in T. cruzi epimastigote cultures. These results suggest that glutathione is an important factor in the resistance of T. cruzi to nifurtimox and benznidazole.

1.Octodon degus shows greater tolerance to pentobarbital as compared with the Wistar rat. 2. Mixed function oxidase activities in liver microsomes were higher in Octodon degus than in the Wistar rat. The reactions assayed were: aminopyrine N-demethylation, aniline and naphthalene hydroxylation and p-nitroanisole O-demethylation. These higher activities seem to be due mainly to the greater cytochrome P-450 content of liver microsomes of Octodon degus. 3. Glutathione S-transferase activity towards 1-chloro-2,4-dinitrobenzene was 30 times higher in Octodon degus than in the Wistar rat. 4. These results may explain the tolerance of Octodon degus to pentobarbital and other drugs.

1. 1. Phosphatase activity in intact Trypanosoma cruzi epimastigotes has been demonstrated. 2. 2. After subcellular fractionation three activities were characterized: (a) a membrane-bound microsomal acid activity with an optimum pH of 4.0 and a K m of 1.2 mM, strongly inhibited by tartrate and fluoride; (b) a soluble cytosolic acid activity with an optimum pH of 5.5 and a K m of 0.95 mM, strongly inhibited by p -hydroxymercuribenzoate, EDTA and copper ions and activated by cyanide, manganese and magnesium ions; and (c) a soluble cytosolic alkaline activity with an optimum pH of 8.0 and a K m of 3.8 mM,inhibited by p -hydroxymercuribenzoate, fluoride, EDTA, and copper, calcium and zinc ions. This activity was increased by magnesium and manganese ions.

T. cruzi epimastigotes have a lysosomal acid phosphatase (pH 4.0) and acid and alkaline phosphatases (pH 5.5 and 8.0) localized in the cytosolic fraction. The levels of the lysosomal acid phosphatase increase with the age of the cultures, but the cytosolic phosphatases decline after the logarithmic phase of growth. The lysosomal phosphatase preferentially hydrolyses low mol. wt phosphate esters; whereas, the cytosolic alkaline phosphatases primarily act on phosphorylated proteins, and both the cytosolic acid and alkaline phosphatases on uridine nucleotide derivatives. The parasite also contains a microsomal glucose 6-phosphatase, and ATPases (Mg2+ and Ca2+-activated) derived from plasma membranes and mitochondria.

1. 1. Trypanosoma cruzi epimastigotes show γ-glutamyltranspeptidase activity which has characteristics significantly different than the mammalian enzyme. 2. 2. The protozoan enzyme is localized in the cytosolic fraction, it has a Km of 1.6 mM and a Vmax of 17.4 nmol/min/mg protein with l-γ-glutamyl-p-nitroanilide as γ-glutamyl donor, and an optimun pH range from 7.5 to 8.0. 3. 3. The best amino acid acceptors were l-histidine, l-asparagine, l-aspartate, l-glutamate and l-proline, but l-glutamine was a very poor acceptor. 4. 4. The enzyme was very sensitive to inhibition by 6-diazo-5-oxo-l-norleucine (k2 = 4.0 × 105M per min) and O-diazo-acetyl-l-serine (k2 = 1.1 × 104M per min). Phenobarbital (k2 = 8.38M per min) and l-serine borate (Ki = 34 mM) were poor inhibitors. 5. 5. The activity of the enzyme was not correlated with the logarithmic phase of growth of the parasites and steadily decreases with the age of the cultures.

Piperonyl Butoxide, Sesoxane, WARF, and DMC were screened as synergists of DDT against Triatoma infestans nymphs. The most effective synergists were found to be the methylene dioxy phenyl compounds, Piperonyl Butoxide, and Sesoxane, which increased the toxicity of DDT by a factor in excess of 20. All the synergists were found to inhibit the oxidative detoxication of 14C-DDT to more polar metabolites. Curiously neither WARF nor DMC inhibited the conversation of DDT to DDE. It is suggested that more effective synergism of DDT will be obtained when suitable inhibitors of T. infestans DDT-dehydrochlorinase are discovered.