Distribution of selenium in sheep treated with dipheny diselenide / Distribuição do selênio em ovinos tratados com disseleneto de difenila

The aim of the present study was to report the in vivo distribution of selenium in sheep. For this, animals were allocated into two groups (control group and treated group) and kept in metabolic cages for a period of 37 days. The treated group received a single dose (6µmol/kg) of Diphenyl Diselenide, intravenously. Plasma and erythrocytes samples were collected at different times. Adipose tissue, muscles (latissimusdorsi, semitendinosus, and supra-scapular) heart, liver, lung, kidney, intestine and brain were sampled at 30 days post-treatment, in order to determine the selenium concentration. The results demonstrated that the selenium, from the Diphenyl Diselenide group, was higher in erythrocytes (4.8mg/L, six hours post-treatment) when compared with the control sheep. The deposition of selenium occurred in the liver (7.01µg/g), brain (3.53µg/g) and kidney (2.02µg/g). After 30 days of a single intravenous injection of Diphenyl Diselenide, liver was the main organ of selenium deposition.(AU)

O objetivo do presente estudo foi investigar a distribuição in vivo do selênio em ovinos. Para isso, os animais foram distribuídos em dois grupos (grupo controle e grupo tratado) e mantidos em gaiolas metabólicas por um período de 37 dias. O grupo tratado recebeu uma dose única (6µmol/kg) de disseleneto de difenila, por via intravenosa. As amostras de plasma e de eritrócitos foram recolhidas em momentos diferentes. Tecido adiposo, músculos (latissimus dorsi, semitendinoso e supraescapular) coração, fígado, pulmão, rim, intestino e cérebro foram amostrados aos 30 dias pós-tratamento, a fim de se determinar a concentração de selênio. Os resultados demonstraram que o selênio, do grupo disseleneto de difenila, foi maior em eritrócitos (4,8mg/L, seis horas após o tratamento) quando comparado com o grupo controle. A deposição de selênio ocorreu no fígado (7,01µg/g), cérebro (3,53µg/g) e rim (2,02µg/g). Após 30 dias de uma única injeção intravenosa de disseleneto de difenila, o fígado foi o principal órgão de deposição de selênio.(AU)

Dexmedetomidine decreases the inflammatory response to myocardial surgery under mini-cardiopulmonary bypass

Cardiopulmonary bypass (CPB) with extracorporeal circulation produces changes in the immune system accompanied by an increase in proinflammatory cytokines and a decrease in anti-inflammatory cytokines. We hypothesize that dexmedetomidine (DEX) as an anesthetic adjuvant modulates the inflammatory response after coronary artery bypass graft surgery with mini-CPB. In a prospective, randomized, blind study, 12 patients (4 females and 8 males, age range 42-72) were assigned to DEX group and compared with a conventional total intravenous anesthesia (TIVA) group of 11 patients (4 females and 7 males). The endpoints used to assess inflammatory and biochemical responses to mini-CPB were plasma interleukin (IL)-1, IL-6, IL-10, interferon (INF)-γ, tumor necrosis factor (TNF)-α, C-reactive protein, creatine phosphokinase, creatine phosphokinase-MB, cardiac troponin I, cortisol, and glucose levels. These variables were determined before anesthesia, 90 min after beginning CPB, 5 h after beginning CPB, and 24 h after the end of surgery. Endpoints of oxidative stress, including thiobarbituric acid reactive species and delta-aminolevulinate dehydratase activity in erythrocytes were also determined. DEX+TIVA use was associated with a significant reduction in IL-1, IL-6, TNF-α, and INF-γ (P<0.0001) levels compared with TIVA (two-way ANOVA). In contrast, the surgery-induced increase in thiobarbituric acid reactive species was higher in the DEX+TIVA group than in the TIVA group (P<0.01; two-way ANOVA). Delta-aminolevulinate dehydratase activity was decreased after CPB (P<0.001), but there was no difference between the two groups. DEX as an adjuvant in anesthesia reduced circulating IL-1, IL-6, TNF-α, and INF-γ levels after mini-CPB. These findings indicate an interesting anti-inflammatory effect of DEX, which should be studied in different types of surgical interventions.

Enzymatic and biochemical characterization of Bungarus sindanus snake venom acetylcholinesterase

This study analyses venom from the elapid krait snake Bungarus sindanus, which contains a high level of acetylcholinesterase (AChE) activity. The enzyme showed optimum activity at alkaline pH (8.5) and 45ºC. Krait venom AChE was inhibited by substrate. Inhibition was significantly reduced by using a high ionic strength buffer; low ionic strength buffer (10 mM PO4 pH 7.5) inhibited the enzyme by 1. 5mM AcSCh, while high ionic strength buffer (62 mM PO4 pH 7.5) inhibited it by 1 mM AcSCh. Venom acetylcholinesterase was also found to be thermally stable at 45ºC; it only lost 5% of its activity after incubation at 45ºC for 40 minutes. The Michaelis-Menten constant (Km) for acetylthiocholine iodide hydrolysis was found to be 0.068 mM. Krait venom acetylcholinesterase was also inhibited by ZnCl2, CdCl2, and HgCl2 in a concentrationdependent manner. Due to the elevated levels of AChE with high catalytic activity and because it is more stable than any other sources, Bungarus sindanus venom is highly valuable for biochemical studies of this enzyme.(AU)

Protective effects of organoselenium compounds against methylmercury-induced oxidative stress in mouse brain mitochondrial-enriched fractions

We evaluated the potential neuroprotective effect of 1-100 µM of four organoselenium compounds: diphenyl diselenide, 3’3-ditri-fluoromethyldiphenyl diselenide, p-methoxy-diphenyl diselenide, and p-chloro-diphenyl diselenide, against methylmercury-induced mitochondrial dysfunction and oxidative stress in mitochondrial-enriched fractions from adult Swiss mouse brain. Methylmercury (10-100 µM) significantly decreased mitochondrial activity, assessed by MTT reduction assay, in a dose-dependent manner, which occurred in parallel with increased glutathione oxidation, hydroperoxide formation (xylenol orange assay) and lipid peroxidation end-products (thiobarbituric acid reactive substances, TBARS). The co-incubation with diphenyl diselenide (100 µM) completely prevented the disruption of mitochondrial activity as well as the increase in TBARS levels caused by methylmercury. The compound 3’3-ditrifluoromethyldiphenyl diselenide provided a partial but significant protection against methylmercury-induced mitochondrial dysfunction (45.4 ± 5.8 percent inhibition of the methylmercury effect). Diphenyl diselenide showed a higher thiol peroxidase activity compared to the other three compounds. Catalase blocked methylmercury-induced TBARS, pointing to hydrogen peroxide as a vector during methylmercury toxicity in this model. This result also suggests that thiol peroxidase activity of organoselenium compounds accounts for their protective actions against methylmercury-induced oxidative stress. Our results show that diphenyl diselenide and potentially other organoselenium compounds may represent important molecules in the search for an improved therapy against the deleterious effects of methylmercury as well as other mercury compounds.

Involvement of glutamate and reactive oxygen species in methylmercury neurotoxicity

This review addresses the mechanisms of methylmercury (MeHg)-induced neurotoxicity, specifically examining the role of oxidative stress in mediating neuronal damage. A number of critical findings point to a central role for astrocytes in mediating MeHg-induced neurotoxicity as evidenced by the following observations: a) MeHg preferentially accumulates in astrocytes; b) MeHg specifically inhibits glutamate uptake in astrocytes; c) neuronal dysfunction is secondary to disturbances in astrocytes. The generation of reactive oxygen species (ROS) by MeHg has been observed in various experimental paradigms. For example, MeHg enhances ROS formation both in vivo (rodent cerebellum) and in vitro (isolated rat brain synaptosomes), as well as in neuronal and mixed reaggregating cell cultures. Antioxidants, including selenocompounds, can rescue astrocytes from MeHg-induced cytotoxicity by reducing ROS formation. We emphasize that oxidative stress plays a significant role in mediating MeHg-induced neurotoxic damage with active involvement of the mitochondria in this process. Furthermore, we provide a mechanistic overview on oxidative stress induced by MeHg that is triggered by a series of molecular events such as activation of various kinases, stress proteins and other immediate early genes culminating in cell damage.

Reaction of diphenyl diselenide with hydrogen peroxide and inhibition of delta-aminolevulinate dehydratase from rat liver and cucumber leaves

The interaction of the product of H2O2 and (PhSe)2 with delta-aminolevulinate dehydratase (delta-ALA-D) from mammals and plants was investigated. (PhSe)2 inhibited rat hepatic delta-ALA-D with an IC50 of 10 æM but not the enzyme from cucumber leaves. The reaction of (PhSe)2 with H2O2 for 1 h increased the inhibitory potency of the original compound and the IC50 for animal delta-ALA-D inhibition was decreased from 10 to 2 æM. delta-ALA-D from cucumber leaves was also inhibited by the products of reaction of (PhSe)2 with H2O2 with an IC50 of 4 æM. The major product of reaction of (PhSe)2 with H2O2 was identified as seleninic acid and produced an intermediate with a lambdamax at 265 nm after reaction with t-BuSH. These results suggest that the interaction of (PhSe)2 with mammal delta-ALA-D requires the presence of cysteinyl residues in close proximity. Two cysteine residues in spatial proximity have been recently described for the mammalian enzyme. Analysis of the primary structure of plant delta-ALA-D did not reveal an analogous site. In contrast to (PhSe)2, seleninic acid, as a result of the higher electrophilic nature of its selenium atom, may react with additional cysteinyl residue(s) in mammalian delta-ALA-D and also with cysteinyl residues from cucumber leaves located at a site distinct from that found at the B and A sites in mammals. Although the interaction of organochalcogens with H2O2 may have some antioxidant properties, the formation of seleninic acid as a product of this reaction may increase the toxicity of organic chalcogens such as (PhSe)2

Influence of the test situation on pup retrieval behavior of normal and undernourished lactating rats

Undernutrition of dams and pups disrupts the retrieval efficiency of mothers. However, if the mothers are assessed in their home cages, they spend more time with their litters. In the present study the effect of test conditions on pup retrieval behavior of mothers receiving a 25 percent (well-nourished group) and 8 percent casein diet (undernourished group) was examined. In agreement with previous studies, undernourished mothers spent more time with their litters than well-nourished dams as lactation proceeded. Pup retrieval behavior varied with test conditions. In the first experiment, the maternal behavior of dams was assessed by the standard procedure (pups were separated from their mother and scattered over the floor of the home cage). The mother was then returned and the number of retrieved pups was recorded. From day 3 to 8, the retrieval efficiency of undernourished dams decreased, while the retrieval efficiency of well-nourished mothers did not vary. In the second experiment, mothers were subjected to a single retrieval test (on day 9 of lactation) using the procedure described for experiment 1. No difference between well-nourished and undernourished mothers was observed. In the third experiment, seven-day-old pups were separated from the mothers and returned individually to a clean home cage. Dietary treatment did not affect the retrieval efficiency. However, undernourished dams reconstructed the nest more slowly than did well-nourished dams. Taken together, these results suggest that pup retrieval behavior of the undernourished mother is not impaired by dietary restriction when the maternal environment is disturbed minimally

Effects of methylmercury exposure during the second stage of rapid postnatal brain growth on negative geotaxis and on delta-aminolevulinate dehydratase of suckling rats

In the present study, we examined the effects of exposure to methylmercury (0, 2.3, 4.6, 6.9 and 9.2 mg/kg, daily for 5 consecutive days, sc) during the second stage of rapid postnatal brain development (8 to 12 days of age) on the sulfhydryl-containing enzyme delta-aminolevulinate dehydratase (ALA-D, E.C. 4.2.1.24) from brain, liver and kidney and on motor performance (latency to complete a negative geotaxis response) of rats. ALA-D specific activity of 13-day old rats of both sexes (7-12 per group) was reduced significantly in rats treated with 6.9 mg/kg and 9.2 mg/kg in brain (about 40 per cent , P < 0.05) and in liver (about 25 per cent , P < 0.05). Renal ALA-D specific activity was not affected by methylmercury treatment. The in vitro IC50 for inhibition of brain, liver and renal ALA-D was 79.3, 81.8 and 39.1 microM, respectively. The latency to complete the negative geotaxis response of 12-day old rats was increased by 6.9 (7.9 +/- 0.7 s, mean +/- SEM) and 9.2 mg/kg methylmercury (7.8 +/- 0.5 s) when compared with control rats (5.8 +/- 0.3 s), suggesting an impairment in motor performance of exposed rats. These results demonstrate that exposure to relatively high doses of methylmercury during the second stage of brain development causes a significant reduction in brain and hepatic ALA-D. The absence of inhibition of ALA-D by lower doses may be related to the relatively low in vitro sensitivity of the enzyme to methylmercury. The possible involvement of ALA-D inhibition on the neurotoxicity of methylmercury deserves additional investigation

Undernutrition during suckling has no effect on the rat locomotor activity response to caffeine

It is known that early malnutrition causes hyposensitivity to serotonergic, gabaergic, catecholaminergic and opioid stimulation. In the present study, we determined whether adult rats undernourished during suckling presented an altered response to caffeine admninistration in a locomotor actiovityyy test. Rats were undernourished during suckling by feeding their dams a 7% casein diet. During the same period, well-nourished dams were fed a 28% casein diet. Animals (90-100 days of age) were habituated to the apparatus. Thereaftert, a dose-response curve for caffeine (2.5, 10.0, 20.0, 40.0 and 120.0 umol/kg, ip) was determined. During handling sessions, undernourished rats presented lower activity scores than well-nourished animals (average values 44.2 ñ 16.4 vs 57.9 ñ 15.4). Well-nourished and undernourished rats responded in a similar way to caffeine administration by increasing the locomotor activity in a dose-dependent manner. Although undernourished animals present an altered sensitivity to various neuropharmacological compounds, the present results indicate that their sensitivity to the locomotor-actiivating effect of caffeine is the same as that of rats well-nourished during suckling

Effects of undernutrition during suckling on foot-shock escape behavior and of post-traini9ng beta-endorphin administration on inhibitory avoidance task test behavior of young rats

The hypothalamic beta-endorphin system of young Wistar rats of both sexes (21-day-old) responds in a distinct way to behavioral situations when compared to adult rats (90 to 120-day-old). In the present study we investigated whether the post-training amnestic effect of beta-endorphin previously demonstrated in Wistar adult rats is also observed young (21-day-old) well-nourished and undernourished rats. rats were undernourished since birth by feeding their dams an 8% casein diet, while well-nourished off spring were fed by dams maintained on a 20% caseindiet beta-endorphin was administered after training in a step-down inhibitory avoidance task using a 0.2 or 0.8 m-A footshock. Retention was tested 24 h later. We observed that the dose of beta-endorphin (1 ug/kg, ip) previously reported to have an amnestic effect on adult rats was ineffective in weanling rats of both nutritional groups. At a higher dose (2 ug/kg, ip) and using a 0.2-mA shock, beta-endorphin impaired the retention only of well-nourished rats. Test-to training difference (in s) in step-down latency for well-nourished beta-endorphin-treated rats was 7 vs 25 s for well-nourished rats treated with saline (P<0.05). Undernourished rats were hyperreactive to this shock intensity. Footshock escape latency (in s) for undernourished rats was 3.56 vs 5.80 for well-nourished rats (P<0.05, experiment 1) and 5.01 vs 10.89 (P<0.05 in experiment 2) and showed better retenmtion than did well-nourished rats. Test-totraining step down latency (in s) for saline-treated undernourished rats was abouth 108 vs 28 saline-treated well-nourished rats (P<0.05). At 0.8-m-A, neither beta-endorphin nor undernutrition affected performance. These resultssuggest that well-nourished rats respons in a distinct way to post-training beta-enmdorphin when compared to adult rats of both sexes. The absence of amnesia in weanling undernourished rats may be related to the enhacenced sensitivity of both sexes. The absence of amnesia in weanling undernourished rats may be related to the enhanced sensitivity of these animals to a 0.2-mA footshock

Effects of undernutrition during suckling on ATP and ADP hydrolysis by synaptosomes from the cerebral cortex of adult rats

Early undernutrition can cause permanent functional changes in the nervous system. Alterations in enzymes involved in neurotransmiter metabolism have been reported to result from early undernutrition. In a previous study, we demonstrated that undernutrition during suckling decreaseATP and ADP hydrolysis by synaptosomes from cerebral cortex by abouth 20% of the value found in 20-day-old well-nourished rats (j.B.T. Rocha, C.F. Melo, J.J.F.Sarkis and R.D. Dias, British Journal of Nutrition, 63:273-283, 1990). In the present study, we investigated whether this deficit persists in synaptosomes from cerebral cortex of nutritionally rehabilitated adult rats. rats were undernourished from birth to 25 days of life by feeding their dams a 7% casein (w/w) diet, while well-nourished offspring were fed by mothers maintained on a 28% casein diet. In contrast to the results previously obtained in young rats, the synaptosomes obtained from the cerebral cortex of early undernourished adult rats hydrolyzed ATP and ADP more efficiently than did those obtained from well-nourished rats. Specific activity (nmol min-1 mg protein-1, mean ñ SD) was 114.9ñ9.5 for undernourished rats (N=8) for ATP, and 50.4ñ6.1 (N=8) vs 38.8ñ4.5 (N=8) for ADP. These results suggest that the deficits found in young rats disappear in rehabilitation adult rats

Long-term treatment with 2,5-hexanedione has no effect on the specific activity of some brain and liver glycolytic enzymes of adult rats

Prolonged exposure to hexacarbon compounds is neurotoxic to humans and animals. As various hexacarbon compounds inhibit glycolytic enzymes in vitro, it has been suggested that this may underlie their neurotoxic effects in vivo. in the present investigation we examined whether long-term treatment with 2,5-hexanedione (200 mg/kg, sc) for 40 days affects the specific activity of brain and liver enolase, lactic dehydrogenase and malate dehydrogenase in female Wistar rats (150-170 g). Glycemia and liver glycogen levels were also determined. The specific activity of all enzymes tested, liver glycogen content and glycemia were not affected by chronic treatment with 2,5-hexanedione. Rats treated with 2,5-hexanedione weighed significantly less than control rats starting on day 18 of treatment (183 ñ 3.4G for the vehicle groups vs 171 ñ 3.2G for the 2,5-hexanedione group). 2,5-hexanedione also increased water intake (46 por cento when compared to vehicle-treated rats). prolonged treatmentof rats with the non-neurotoxic hexacarbon 1,6-hexanediol (207 mg/kg, sc) significantly increased liver glycogen content (5.9 ñ 0.6g/100g for the vehicle group vs 9.0 ñ 1.1g/100 g for the 1.6-hexanediol group) as well as food intake (44.0 ñ 1.5g 100g-1 6 days-1 for thge 1,6-hexanediol group). These results indicate that long-term treatment with 2,5-hexanedione did not alter the brain and liver glycolytic enzymes studied, liver glycogen content or glycemia but did reduce weight gain and increased water intake, whereas the administration of the reportedly non-neurotoxic hexacarbon 1,6-hexanediol has demonstrable metabolic effects

Ontogeny of ATP and ADP hydrolysis by cerebral cortex synaptosomes from rats

In the present study, we examined the ontogeny of ATP and ADP hydrolysis by cerebral cortex symptosomes from rats of various ages (0-, 7-, 14-, 21- and 60 to 90-day-old rats) in order to learn whether hydrolytic activity increases during the period of intense brain grwth, as has been reported for other enzymes involved in neurotransmitter metabolism. the results demonstrate that ATP and ADP hydrolyzing activities increase in parallel from birth until the second postnatal week (about 4-fold), followed by a slight and statistically insignificant increase until the animal reaches adulthood. The maximum increase in nucleotide hidrolysis coincided with mximum brain growth, which may indicate a role for the enzyme in neurotransmission. Furthermore, the parallel development of both activities (ATPase and ADPase) strongly suggest that a single enzyme, an ATP diphosphohydrolase, is involved in ATP and ADP hydrolisis by the synaptosomal fraction

Effects of chronic treatment with high doses of chlorpromazine on ATP and ADP hydrolysis by synaptosomal fractions from the rat caudate nucleus

Several studies have indicated that chlorpromazine and its metabolites affect ATP hydrolysis by brain and liver plasma membranes in vitro. The present report examines whether chronic treatment (12 days) with high doses of chlorpromazine (10 and 40 mg/kg) could affect ATP and ADP hydrolysis by synaptosomal fractions from the rate caudate nucleus. Both doses of chlorpromazine caused significant and paralled decreases (23 to 31%) in the ATP and ADP hydrolysis. The parallelism between the effects of chlorpromazine on ATP and ADP hydrolys suggests the participation of a single enzyme (ATP diphosphohydrolase) in nucleotide hydrolysis

Effect of caffeine adminsitration on latent learning ability of male rats in a simple maze task

Animals can acquire information about the environment in the absence of conventional rewards. This latent or incidental learning depends on their interaction with the environment. The present report examines whether caffeine (120 micron mol/kg) could enhance latent learning of a simple maze task by increasing exploratory activity of rats in the maze. Though caffeine increased the activity of rats in the maze under a nonappetitive condition, methylxanthine did not improve performance in rats tested under an appetitive condition in the maze. The results suggest that caffeine (120 micron mol/kg) does not facilitate latent learning of a simple maze task. An interesting and unexpected finding of the present study was that caffeine caused greater stimulation of exploratory activity in the open-field than in the maze under nonappetitive conditions. This may be due to differences in the complexity of the two paradigms

Undernutrition during suckling does not change the specific or total activity of hypothalamic proline endopeptidase in adult rats

Undernutrition during suckling causes a decrease in hypothalamic beta-endorphin-like immunoreactivity in rats. Since proline endopeptidase (E.C. 3.4.21.26) has been proposed to play a role in the processing of Beta-endorphin, we examined the effects of undernutrition during suckling on the enzyme activity. Rats were undernourished by feeding their dams an 8% casein diet from the day of birth until weaning (21 days). Dams of well-nourished rats were fed a 25% casein diet during the same period. After weaning, all rats received a 20% protein diet until 90 to 120 days of age when they were killed for the enzyme assay. The specific and total activity of hypothalamic proline endopeptidase was not altered by undernutrition followed by nutritional rehabilitation(2.37 + or - 0.24 nmol sulphamethoxazole min-1 mg-1 for well-nourished rats vs 2.68 + or - 0.24 nmol sulphamethoxazole min-1 mg-1 for undernourished rats). This lack of correlation suggests that proline endopeptidase is probably not responsible for the low levels of hypothalamic Beta-endorphin found in adult rats submitted to undernutrition during suckling

Effects of undernutrition during suckling and of post-training ß-endorphin administration on avoidance performances of adult rats

1. The effects of undernutrition during suckling and of post-training ß-endorphin administration on avoidance task were invstigated in adult rats. 2. young rats were undernourished from delivery until weaning (21 days) by feeding their mothers a diet conatining 8% protein (w/w). Mothers of well-nourished rats were fed a 20% protein diet. After weaning, both groups of rats were fed a 20% protein diet until 90-120 days if age, when they were subjected to behavioral sessions. 3. Acquistion was measured in training sessions and retention in test sessions 24 h after training. Beta-endorphin or salina (control) was injected ip immdiately after training. Rats were subjected to shuttle and step-down inhibitory avoidance sessions using footshock of 0.2 or 0.8 mA intensity. 4. Undernutrition during suckling caused hyperreactivity to 0.2 mA footshocks. Beta-endorphin caused amnesia to shuttle avoidance task only in normal rats trained with 0.8 mA. Foor-shocks. In the step-down inhibitory avoidance task, ß-endorphin was amnesic only for normal rats and only for 0.2-mA footshocks. Beta-endorphin was not amnesic in undernourished rats