Toxicity evaluation of nanocrystalline silver-impregnated coated dressing on the life cycle of worm Caenorhabditis elegans.

In recent times, however, due to the emergence of bacterial strains with resistance to conventional antibiotics, silver has again gained attention as an alternative for developing new efficient bactericides, including the use of silver nanoparticles (AgNPs). However, the improper disposal of these items after use may cause toxicological effects on organisms in the environment. To evaluate the potential environmental hazard of nanosilver-coated dressings, the nematode Caenorhabditis elegans was chosen as a test organism. The assays were conducted in 24-well plates that contain four different sizes of coated dressing to obtain different concentrations. L1 and L4 C. elegans larval stages were exposed to these nanosilver concentrations. Dressing cutouts were arranged between two layers of agar for 3 days and Escherichia coli (OP 50 strain) was added as food source for the worms. After the exposure period, growth, reproduction, fertility, silver concentration in the medium and the concentration of reactive oxygen species (ROS) in the worms were evaluated. Scanning and transmission electron microscopy analyses were performed on the coated dressings, as well as analyses of zeta potential, ionic release and antibacterial power in two bacterial strains (Pseudomonas aeruginosa and Staphylococcus aureus). It was verified the antibacterial power of the coated dressing, in both bacteria strains tested. Characterization of the coated dressing indicated heterogeneous nanoparticles, as well as distinct zeta potentials for the medium in water and saline medium (0.9% NaCl). L1 larval worms exposed to nanosilver-coated dressing showed a high ROS concentration and reductions in growth, fertility and reproduction. Worms exposed to the coated dressing during the L4 stage showed almost no response. Overall, the obtained results indicate the potential environmental hazard of nanosilver-coated dressings.

Neurotoxicity in zebrafish exposed to carbon nanotubes: Effects on neurotransmitters levels and antioxidant system.

Given the increasing use of carbon nanotubes (CNT) in several industries and technological applications, it is essential to perform in vivo toxicological studies with these nanomaterials to evaluate their potential ecotoxicity. Dopamine (DA) and serotonin (5HT) are key neurotransmitters for brain functions and behavioral responses. Determination of DA and 5HT were performed in brain samples from zebrafish Danio rerio exposed i.p. to single-walled CNT (SWCNT), besides analyzing acetylcholinesterase (AChE) and ectonucleotidases activity, lipid peroxidation and total antioxidant capacity. Results showed that treatment with SWCNT increased between 3 and 6-fold the concentration of DA and 5HT (p < 0.05). Similarly, a significant reduction (p < 0.05) in AChE activity was observed in the brains of SWCNT exposed zebrafish when compared to the control groups. Cholinergic, serotonergic, and dopaminergic systems, through AChE activity and serotonin and dopamine levels, respectively were affected by SWCNT in the zebrafish brain. Alterations in these neurotransmitters can potentially affect several physiological and behavioral that they control.

Interference of single walled carbon nanotubes (SWCNT) in the measurement of lipid peroxidation in aquatic organisms through TBARS assay.

Nanomaterials (NM) exhibit unique properties due their size and relative area, but the mechanisms and effects in the living organisms are yet to be unfold in their totality. Potential toxicity mechanisms concerning NM as carbon nanotubes include oxidative stress generation. Several fluorimetric and colorimetric methods have been systematically used to measure NM toxicity, and controversial results have been reported. One of the problems can be related to the interference effects induced by NM, leading to artifacts that can lead to misleading conclusions. In present study, it was performed in vitro assays with two aquatic species: the zebrafish Danio rerio and the polychaete Laeonereis acuta to evaluate the potential interference capacity of single-wall carbon nanotubes (SWCNT) in a fluorometric method (TBARS assay) to measure lipid peroxidation. Obtained results indicated that gills and brain of zebrafish presented a lowered fluorescence only at extremely high concentrations (50 and 500mg/L). Determinations in anterior, middle, and posterior body regions of L. acuta showed a quite different pattern: high fluorescence at low SWCNT concentrations (0.5mg/L) and lowering at the highest (500mg/L). To eliminate matrix effect of biological samples, tests employing the standard for TBARS assay, 1,3,3-tetramethoxipropane, were run and the results showed again higher fluorescence values at low concentrations (0.5-5mg SWCNT/L), a technique artifact that could lead to misleading conclusions since higher fluorescence values implicate higher TBARS concentration, implying oxidative stress. Using the colorimetric FOX assay with cumene hydroperoxide as standard presented remarkable better results since no artifacts were observed in the same SWCNT concentration range that employed with the TBARS technique.

Oxidative status of the myocardium in response to different intensities of physical training.

The intensity of exercise determines the metabolic pathway and the energetic substrate that is spent. Our study sought to identify the effects of different intensities of swimming on myocardial oxidative status and the blood lipid profile. Eighty Wistar rats (male and female) submitted to different intensities of a swimming regimen (low, LS; moderate, MS; or high, HS) for 16 weeks. Samples of blood and myocardium from the left ventricle were collected to determine lipid profiles and oxidative status. Reactive oxygen species (ROS) and antioxidant capacity against peroxyl radicals (ACAP), lipid profiles and lipid peroxidation was analyzed. ROS levels and ACAP were higher in male rats than in female rats overall (p<0.05). However, ACAP in the myocardium was significantly elevated in LS female rats compared to the MS and HS female rats, which had a significantly lower ACAP compared to all other groups. LS and MS training in both sexes and HS training (in females) led to significant decreases in the heart's lipid peroxidation. Amelioration of the lipid profile and reduction in oxidative damage contributed to a physiological state that benefits cardiovascular function in exercised animals. The results show that low and moderate intensity exercise promotes beneficial adaptations.

Effects of lipoic acid on growth and biochemical responses of common carp fed with carbohydrate diets.

Lipoic acid (LA) is an antioxidant that also favors glucose uptake in mammals. Until now, there are no studies evaluating the potential effect of this molecule on glycemic control in fish. It was evaluated LA effects on glucose uptake in common carp Cyprinus carpio fed with carbohydrate diets from two carbohydrate sources: glucose (GLU) and starch (STA), and supplemented or not with LA, being the diets: +GLU/-LA (GLU); +GLU/+LA (GLU + LA); +STA/-LA (STA); and +STA/+LA (STA + LA). Carp juveniles (6.5 ± 0.1 g) were fed with each diet ad libitum 4 times a day, during 68 days. Muscle glycogen concentration was higher (p < 0.05) in GLU and GLU + LA than in STA and STA + LA groups. On fish fed with starch, muscle cholesterol and triglyceride concentrations were higher (p < 0.05) in fish fed diets supplemented with LA. Muscle protein levels were higher in fish fed with LA, independent of the diet carbohydrate source. Lipid peroxidation was significantly reduced (p < 0.05) in fish muscle on fish fed the STA + LA diets when compared with the STA diet. Our findings indicate that LA modulates lipid, proteins and carbohydrate metabolism together with the well-known antioxidant effect. Also, LA showed to enhance starch utilization taking into account muscle cholesterol and triglyceride levels.

Uptake, metabolism and sub-lethal effects of BDE-47 in two estuarine invertebrates with different trophic positions.

Two microcosm types -sediment-biota and biota-biota- were constructed to simulate different pathways of BDE-47 uptake, metabolism and oxidative stress effects in two key estuarine invertebrates (polychaete Laeonereis acuta and crab Cyrtograpsus angulatus). In the sediment-biota experiment, both species were exposed to spiked sediments; an environmentally reported and a high concentration of BDE-47 for 2 weeks. In the biota-biota experiment, crabs were fed with polychaetes pre-exposed to BDE-47 in the sediment-biota experiment. The sediment-biota experiment first revealed that polychaetes significantly accumulated BDE-47 (biota-sediment accumulation factor >2; p < 0.05) to a much greater extent than the crab organs (muscle, hepatopancreas, gills) at both sediment concentrations. For oxidative stress responses, polychaete and crab tissues exposed to spiked sediment showed a significant increase (p < 0.05) of only glutathione S-transferase (GST) activity with respect to controls in both BDE-47 concentrations. No lipid peroxidation (TBARS) or total antioxidant capacity (ACAP) changes were evident in the species or organs exposed to either BDE-47 sediment concentration. The biota-biota experiment showed that feeding crabs with pre-exposed polychaetes caused BDE-47 accumulation in organs as well as significant amounts of BDE-47 eliminated through feces (p < 0.05). Unlike the sediment-biota exposure, crabs fed with pre-exposed BDE-47 polychaetes showed the most conspicuous oxidative stress responses. Significant changes in GST and ACAP in both hepatopancreas and gills, in addition to enhanced TBARS levels in the hepatopancreas with respect to controls (p < 0.05), revealed that BDE-47 assimilated by invertebrates represents a potential source of toxicity to their predators. No methoxylated metabolites (MeO-PBDEs) were detected during BDE-47 metabolism in the invertebrates in either of the two different exposure types. In contrast, hydroxylated metabolites (OH-PBDEs) were detected in polychaetes and crab organs/feces in both experiments. Our results demonstrate that PBDE hydroxylation is one of the main biotransformation routes of BDE-47 in estuarine animals, which could be associated with the oxidative stress responses found.

Age protects from harmful effects produced by chronic intermittent hypoxia.

Obstructive sleep apnoea (OSA) affects an estimated 3­7% of the adult population, the frequency doubling at ages >60­65 years. As it evolves, OSA becomes frequently associated with cardiovascular, metabolic and neuropsychiatric pathologies defining OSA syndrome (OSAS). Exposing experimental animals to chronic intermittent hypoxia (CIH) can be used as a model of the recurrent hypoxic and O2 desaturation patterns observed in OSA patients. CIH is an important OSA event triggering associated pathologies; CIH induces carotid body (CB)-driven exaggerated sympathetic tone and overproduction of reactive oxygen species, related to the pathogenic mechanisms of associated pathologies observed in OSAS. Aiming to discover why OSAS is clinically less conspicuous in aged patients, the present study compares CIH effects in young (3­4 months) and aged (22­24 months) rats. To define potential distinctive patterns of these pathogenic mechanisms, mean arterial blood pressure as the final CIH outcome was measured. In young rats, CIH augmented CB sensory responses to hypoxia, decreased hypoxic ventilation and augmented sympathetic activity (plasma catecholamine levels and renal artery content and synthesis rate). An increased brainstem integration of CB sensory input as a trigger of sympathetic activity is suggested. CIH also caused an oxidative status decreasing aconitase/fumarase ratio and superoxide dismutase activity. In aged animals, CIH minimally affected CB responses, ventilation and sympathetic-related parameters leaving redox status unaltered. In young animals, CIH caused hypertension and in aged animals, whose baseline blood pressure was augmented, CIH did not augment it further. Plausible mechanisms of the differences and potential significance of these findings for the diagnosis and therapy of OSAS are discussed.

Crescimento e metabolismo do nitrogênio em juvenis de Trachinotus marginatus alimentados com diferentes níveis proteicos / Growth and nitrogenous metabolism of juveniles Trachinotus marginatus feeding with differently protein levels

O objetivo deste trabalho foi avaliar o crescimento e alterações no metabolismo do nitrogênio, além da composição do fígado e do músculo de juvenis de pampo Trachinotus marginatus alimentados com 43, 54 ou 64% de proteína bruta (PB43, PB54 e PB64, respectivamente). Grupos de 10 juvenis (1,47±0,14g) foram estocados em nove tanques de 50L, em sistema de recirculação de água marinha, durante 55 dias, alimentados quatro vezes ao dia com as rações pré-definidas. O ganho em peso, a taxa de crescimento específico e a conversão alimentar não apresentaram diferença significativa (P>0,05). Contudo, o acréscimo de proteína consumida aumentou o índice hepatossomático e a taxa de excreção de amônia pós-prandial, bem como diminuiu a retenção proteica. Os peixes alimentados com a dieta PB43 apresentaram menor concentração da transaminase glutâmico oxalacética hepática, além de menor concentração de triglicerídeos no músculo e fígado (P<0,05). A composição da carcaça, o teor de glicogênio e de proteínas totais para músculo e fígado não mostraram diferenças significativas (P>0,05), excetuando o maior teor de lipídios da carcaça no tratamento PB43. Concluímos que a melhor utilização da proteína por juvenis de pampo é observada quando eles são alimentados com uma dieta contendo 43% de proteína.

The experiment aimed to evaluate the growth, the nitrogen metabolism and liver and muscle composition of juvenile pompano Trachinotus marginatus fed with different protein levels (PB43, PB54, PB64). Groups of 10 juveniles (1.47±0.14g) were stocked in nine tanks with 50L in a marine recirculating aquaculture system during 55 days. Fish were fed four times a day. The weight gain, specific growth rate and feed conversion rate did not present significant differences (P>0.05). However, the increase in protein consumption augmented the hepatosomatic index, the pos prandial ammonia excretion rate and reduced the protein efficiency rate. Fish fed with PB43 presented lower hepatic glutamic oxaloacetic transaminase and lower concentration of triglycerides in muscle and liver (P<0.05). Carcass composition, glycogen and muscle and liver protein content did not present significant differences (P>0.05), with the exception of the lipid content in fish carcass fed with PB43. It is concluded that the better protein utilization in juvenile pompano is observed when they are fed with 43% protein.

Antioxidant and toxicological effects elicited by alpha-lipoic acid in aquatic organisms.

Lipoic acid (LA) is a disulfide-containing compound derived from octanoic acid that is synthesized in mitochondria. This molecule acts as a co-factor for mitochondrial enzymes that catalyze oxidative decarboxylation reactions. Several antioxidant properties of LA enable it to be considered as an "ideal antioxidant", having diverse benefits that allow it to deal with environmental or biological stress. Some of the effects induced by LA in aquatic organisms render it suitable for use in aquaculture. However, it is necessary to determine the appropriate dose(s) to be used with different species and even organs to maximize the beneficial antioxidant and detoxifying effects and to minimize the pro-oxidant toxic effects. This review analyzes and compiles existing data from aquatic organisms in which both benefits and drawbacks of LA have been described.

Antioxidant, phase II and III responses induced by lipoic acid in the fish Jenynsia multidentata (Anablapidae) and its influence on endolsulfan accumulation and toxicity.

Antioxidants like lipoic acid (LA) are known to trigger augmented antioxidant and phase II and III responses. This study aimed to evaluate the effect of LA in P-glycoprotein (Pgp) expression, glutathione-S-transferase (GST) activity, total antioxidant competence, levels of lipid peroxides (TBARS) and accumulation of the organochlorine insecticide endosulfan (Endo: α-, ß-isomers and sulfate metabolite) in different organs of the fish Jenynsia multidentata. One hundred and twenty females (1.55±0.07 g) were fed during 8 days with (n=60) or without (n=60) a LA enriched ration (6000 mg/kg). Four experimental groups were defined: -LA/-Endo; +LA/-Endo; -LA/+Endo; and +LA/+Endo. Endo groups were exposed during 24 h to 1.4 µg of insecticide/L. Results showed that only LA induced a significant increment in liver Pgp expression. GST activity was augmented in liver after exposure to LA or Endo. TBARS levels were lowered in liver and gills after LA pre-treatment. Total antioxidant capacity was lowered in liver of Endo exposed fish, a result that was reversed by LA pre-treatment. It is concluded that LA induced the expected effects in terms of Pgp expression, GST activity and reduced TBARS levels although favored α-Endo accumulation in brain. However, the Endo metabolism to the more persistent endosulfan sulfate was not facilitated by LA pre-treatment.

Biochemical responses and physiological status in the crab Hemigrapsus crenulatus (Crustacea, Varunidae) from high anthropogenically-impacted estuary (Lenga, south-central Chile).

Estuarine environmental assessment by sub-individual responses is important in order to understand contaminant effects and to find suitable estuarine biomonitor species. Our study aimed to analyze oxidative stress responses, including glutathione-S-transferase (GST) activity, total antioxidant capacity (ACAP) and lipid peroxidation levels (TBARS) in estuarine crabs Hemigrapsus crenulatus from a high anthropogenically-impacted estuary (Lenga) compared to low and non-polluted estuaries (Tubul and Raqui), in a seasonal scale (winter-summer), tissue specific (hepatopancreas and gills) and sex related responses. Results showed that hepatopancreas in male crabs better reflected inter-estuary differences. Morpho-condition traits as Cephalothorax hepatopancreas index (CHI) could be used as an indicator of physiological status of estuarine crabs. Discriminant analysis also showed that GST and TBARS levels in summer are more suitable endpoints for establishing differences between polluted and non-polluted sites. These results suggest the importance of seasonality, target tissue, sex and physiological status of brachyuran crabs for estuarine biomonitoring assessment.

Integrated biological responses of zebrafish (Danio rerio) to analyze water quality in regions under anthropogenic influence.

This study analyzed water quality in regions around Patos lagoon (Southern Brazil) that are under anthropogenic pressure. Water samples were collected from five different sites, including one used as a source for human consumption (COR) and others known to be influenced by human activities (IP). Danio rerio (Teleostei, Cyprinidae) organisms were exposed for 24h to these water samples, plus a control group. It was observed that: (1) reactive oxygen species levels were lower in COR and IP than in the control group; (2) glutamate-cysteine ligase (catalytic subunit) expression was higher in COR than in other sites; (3) exposure to all water samples affected long-term memory (LTM) when compared to control group. Thus, some water samples possess the ability to modulate the antioxidant system and to induce a decline in cognitive functions, as measured by LTM. The obtained results indicate that a combination of variables of different organization level (molecular, biochemical and behavioral) can be employed to analyze water quality in impacted regions.

Acetylcholinesterase activity and antioxidant capacity of zebrafish brain is altered by heavy metal exposure.

Pollution is a world problem with immeasurable consequences. Heavy metal compounds are frequently found as components of anthropogenic pollution. Here we evaluated the effects of the treatment with cadmium acetate, lead acetate, mercury chloride, and zinc chloride in acetylcholinesterase activity and gene expression pattern, as well as the effects of these treatments in antioxidant competence in the brain of an aquatic and well-established organism for toxicological analysis, zebrafish (Danio rerio, Cyprinidae). Mercury chloride and lead acetate promoted a significant decrease in acetylcholinesterase activity whereas they did not alter the gene expression pattern. In addition, the antioxidant competence was decreased after exposure to mercury chloride. The data presented here allowed us to hypothesize a signal transmission impairment, through alterations in cholinergic transmission, and also in the antioxidant competence of zebrafish brain tissue as some of the several effects elicited by these pollutants.

Biomonitoring of antioxidant and oxidative stress responses in Perinereis gualpensis (Polychaeta: Nereididae) in Chilean estuarine regions under different anthropogenic pressure.

This study aimed to analyze oxidative stress parameters, including levels of the antioxidant glutathione (GSH), activity of glutamate-cysteine ligase (GCL) and glutathione-S-transferase (GST), total antioxidant capacity and protein oxidation, in the polychaete Perinereis gualpensis (Nereididae) collected from the Biobío, Itata, Valdivia and Lingue estuaries in Chile, which present different degrees of anthropogenic pressure. Sampling sites were characterized considering a geographic information system and the physicochemical characteristics of water and sediment. Significant differences (p<0.05) were observed between the sampling sites for most of the responses (GSH, GCL, GST and antioxidant capacity), mainly related to human activities such as agriculture, industry, among others. Multivariate correlation analysis indicates a certain relationship of antioxidant responses with human activities, salinity, and worm weight, this last employed to standardize GST and antioxidant capacity. These results clearly indicate biomarker responses in P. gualpensis in Biobío and Valdivia estuaries, the more affected by human activities.

Oxidative stress generation by microcystins in aquatic animals: why and how.

Microcystins (MICs) are potent toxins produced worldwide by cyanobacteria during bloom events. Phosphatases inhibition is a well recognized effect of this kind of toxins as well as oxidative stress. However, it is not fully understood why and how MICs exposure can lead to an excessive formation of reactive oxygen species (ROS) that culminate in oxidative damage. Some evidences suggest a close connection between cellular hyperphosphorylation state and oxidative stress generation induced by MICs exposure. It is shown, based on literature data, that MICs incorporation per se can be the first event that triggers glutathione depletion and the consequent increase in ROS concentration. Also, literature data suggest that hyperphosphorylated cellular environment induced by MICs exposure can modulate antioxidant enzymes, contributing to the generation of oxidative damage. This review summarizes information on MICs toxicity in aquatic animals, focusing on mechanistic aspects, and rise questions that in our opinion needs to be further investigated.

Effect of silymarin on biochemical parameters of oxidative stress in aged and young rat brain.

Silymarin (SM), the active complex of milk thistle, is a lipophilic fruit extract and is composed of several isomer flavonolignans. Flavonoids are antioxidants found molecules capable of intercepting reactive oxygen species (ROS). The oxidative stress (OS) is caused by imbalance between antioxidant defenses and production of ROS causing oxidative damage to macromolecules. Brain is susceptible to oxidative stress and it is associated with age-related brain dysfunction. This study evaluated the effect of SM on biochemical parameters that evaluate OS in aged and young rat brain. For measures of OS were used measures of total oxyradical scavenging capacity (ACAP) through the concentration of ROS by fluorescence, lipid peroxidation (LPO), via FOX and TBARS, proteins oxidation by Western blot (WB). Rats were treated with SM at doses of 200 and 400mg/kg/day (SM200 and SM400). The LPO analyzed through FOX was increased in the hippocampus of aged animals treated with SM400, but in the cortex of young and aged, the highest dose of SM decreased LPO analyzed through TBARS. Both doses have seemed most effective in the reduction of oxidized proteins in aged brain. These results suggest that SM may contribute to the prevention of aged-related and pathological degenerative processes in the brain.

Seasonal and organ variations in antioxidant capacity, detoxifying competence and oxidative damage in freshwater and estuarine fishes from Southern Brazil.

This study analyzed biochemical biomarkers of freshwater and estuarine fish species from Southern Brazil. It analyzed three organs (muscle, liver and gills), in four fish species (Micropogonias furnieri, Pimelodus pintado, Loricariichthys anus and Parapimelodus nigribarbis) in order to perform an environmental diagnosis. Obtained results showed that liver of L. anus and gills of M. furnieri presented higher total antioxidant capacity against peroxyl radicals during fall, whereas a clear seasonality was found for gill reduced glutathione (GSH) levels of all studied species, with higher concentration during spring. In terms of oxidative damage (TBARS), liver of M. furnieri and gills of P. nigribarbis showed higher TBARS levels during fall, whereas P. pintado showed the lowest TBARS value. Finally, a conspicuous seasonal effect was observed for purified and non-purified glutathione-S-transferase (GST), where minimum values were registered during fall, pointing to this season as one where fish species are less competent to perform detoxifying reactions.

Microcystin-induced oxidative stress in Laeonereis acuta (Polychaeta, Nereididae).

Oxidative stress induced by microcystins was evaluated in an estuarine worm, Laeonereis acuta (Nereididae). Ten organisms were exposed to lyophilized cells of a toxic Microcystisaeruginosa strain RST9501 ( approximately 2 microg/mL microcystins, MC); 10 were exposed to lyophilized cells of a nontoxic Aphanotece sp. strain RSMan92 and 10 were maintained without cyanobacterial cells. Exposure time was 48 h. The enzymatic antioxidant defenses, as well as the oxidative damage, were analyzed. Toxic and nontoxic cyanobacteria lowered catalase activity with no changes in glutathione reductase and glutathione-S-transferase activities. This may have led to toxin intracellular accumulation, which should favor oxidative stress generation, observed by the high lipid peroxide and DNA-protein crosslink levels in the group exposed to MC.

Estradiol valerate and tibolone: effects upon brain oxidative stress and blood biochemistry during aging in female rats.

Estrogen compounds have been described as important brain protectors. This study investigated the effects of estradiol valerate (EV--0.3 mg/kg) and two concentrations of tibolone (TB1=0.5 mg/kg and TB2=1 mg/kg) on brain oxidative stress parameters and blood biochemistry in ovariectomized female rats, of three different age groups (young--2 months, adult--8 months, and old--20 months). In the brain cortex, young and old TB2-treated and old no-hormone-replacement (NR) females showed lower lipid hydroperoxide (LPO) levels compared to young Sham and adult TB1 animals (P<0.05). Also in the cortex, both tibolone doses produced higher (P<0.05) total antioxidant capacity (TOSC) levels compared to EV-treated adult females. Ovariectomized adult females (NR, EV, TB1 and TB2) showed lower (P<0.05) TOSC levels in the hippocampus compared to the Sham control. Reactive oxygen species (ROS) were higher (P<0.05) in old females compared to all younger ones. TB2-treated adults showed higher plasma glucose (P<0.05) levels compared to old animals. Regardless of age, TB2 treatment increased female (P<0.05) LDL levels compared to Sham and EV-treated animals. In old females, TB2 significantly increased HDL levels compared to Sham controls, and decreased triglyceride levels were shown in EV, TB1 and TB2 compared to Sham old females. The Atherogenic Index of Plasma was higher (P<0.05) in adult tibolone-treated females compared to both young and old TB2-treated females. These results suggest that the effects of gonad steroid on brain and blood physiology change significantly with aging, and that evaluating hormonal treatment types and doses could be the key factor in the potential use of a specific hormone therapy.

Antioxidant responses of Laeonereis acuta (Polychaeta) after exposure to hydrogen peroxide.

The effects of H2O2 were evaluated in the estuarine worm Laeonereis acuta (Polychaeta, Nereididae) collected at the Patos Lagoon estuary (Southern Brazil) and maintained in the laboratory under controlled salinity (10 psu diluted seawater) and temperature (20 degrees C). The worms were exposed to H2O2 (10 and 50 microM) for 4, 7, and 10 days and the following variables were determined: oxygen consumption, catalase (CAT) and glutathione peroxidase activity in both the supernatant and pellet fractions of whole body homogenates. The concentrations of non-protein sulfhydryl and lipid peroxides (LPO) were also measured. The oxygen consumption response was biphasic, decreasing after 4 days and increasing after 7 and 10 days of exposure to 50 microM H2O2 (P < 0.05). At the same H2O2 concentration, CAT activity was lower (P < 0.05) in the pellet fraction of worms exposed for 10 days compared to control. Non-protein sulfhydryl concentration and glutathione peroxidase activity were not affected by H2O2 exposure. After 10 days, LPO levels were higher (P < 0.05) in worms exposed to 50 microM H2O2 compared to control. The reduction in the antioxidant defense was paralleled by oxidative stress as indicated by higher LPO values (441% compared to control). The reduction of CAT activity in the pellet fraction may be related to protein oxidation. These results, taken together with previous findings, suggest that the worms were not able to cope with this H2O2 concentration.