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Açaí (Euterpe oleracea) has a rich nutritional composition, showing nutraceutical and protective effects in several organs. In this study, the effects of an açaí-enriched diet on motor performance, anxiety-like behavior, and memory retention were deeply investigated.Eight-week male Wistar rats were fed with an Euterpe oleracea (EO) pulp-enriched diet, an olive oil-enriched (OO) diet (polyunsaturated fatty acid [PUFA] fat control diet), or a chow diet for 31 days (28 days pre-treatment and 3 days during behavioral tests). Afterward, animals were submitted to a battery of behavioral tests to evaluate spontaneous motor behavior (open-field test), anxiety-like behavior (elevated plus maze and open-field test), and memory retention (step-down). Oxidative stress in the hippocampus was evaluated by a lipid peroxidation assay.EO-enriched diet did not influence the body weight and food intake but increased the glucose plasmatic level after 31 days under this diet. However, a similar fat-enriched diet stimulated a marked weight gain and reduced the food intake, followed by changes in the plasmatic lipid markers. EO-enriched diet preserved the motor spontaneous performance, increased the exploration in the aversive environment (anxiolytic-like effects), and elevated the latency to step-down (improved memory retention). The EO-enriched diet also reduced the level of lipid peroxidation in the hippocampus. These positive effects of EO-enriched diet can greatly support the usage of this diet as a preventive therapy.Taken together, the current study suggests that Euterpe oleracea-enriched diet promotes anxiolytic-like effects and improves memory consolidation, possibly due to the reduced levels of lipid peroxidation in the hippocampus.

To investigate the magnitude and time course of pseudorandom ffERG during light adaptation. Ten healthy subjects (26 ± 10.1 years) underwent 20 min of dark adaptation, and then the ffERG was evoked by pseudorandom flash sequences (4 ms per flash, 3 cd.s/m2) driven by m-sequences (210–1 stimulus steps) using Veris Science software and a Ganzfeld dome over a constant field of light adaptation (30 cd/m2). The base period of the m-sequence was 50 ms. Each stimulation sequence lasting 40 s was repeated at 0, 5, 10, 15 and 20 min of light adaptation. Relative amplitude and latency (corrected by values found at 0 min) of the three components (N1, P1, and N2) of first-order (K1) and first slice of the second-order (K2.1) kernel at 5 time points were evaluated. An exponential model was fitted to the mean amplitude and latency data as a function of the light adaptation duration to estimate the time course (τ) of the light adaptation for each component. Repeated one-way ANOVA followed by Tukey post-test was applied to the amplitude and latency data, considering significant values of p

Current theories on the role of serotonin (5-HT) in vertebrate defensive behavior suggest that this monoamine increases anxiety but decreases fear, by acting at different levels of the neuroaxis. This paradoxical, dual role of 5-HT suggests that a serotonergic tone inhibits fear responses, while an acute increase in 5-HT would produce anxiety-like behavior. However, so far no evidence for a serotonergic tone has been found. Using zebrafish alarm responses, we investigate the participation of phasic and tonic 5-HT levels in fear-like behavior, as well as in behavior after stimulation. Conspecific alarm substance (CAS) increased bottom-dwelling and erratic swimming, and animals transferred to a novel environment after CAS exposure (post-exposure behavior) showed increased bottom-dwelling and freezing. Clonazepam blocked CAS effects during and after exposure. Acute fluoxetine dose-dependently decreased fear-like behavior, but increased post-exposure freezing. Metergoline had no effect on fear-like behavior, but blocked the effects of CAS on post-exposure behavior; similar effects were observed with para-chlorophenylalanine. Finally, CAS was shown to decrease the activity of monoamine oxidase in the zebrafish brain after exposure. These results suggest that phasic and tonic serotonin encode an aversive expectation value, switching behavior toward cautious exploration/risk assessment/anxiety when the aversive stimulus is no longer present.

Acute stress is an important factor in the development of anxiety disorders. Zebrafish are an organism model widely used by studies that aim to describe the events in the brain that control stress-elicited anxiety. The goal of the current study was to evaluate the pattern of cell activation in the telencephalon of adult zebrafish and the role of the GABAergic system on the modulation of anxiety-like behavior evoked by acute restraint stress. Zebrafish that underwent acute restraint stress presented decreased expression of the c-fos protein in their telencephalon as well as a significant decrease in GABA release. The data also supports that decreased GABA levels in zebrafish brains have diminished the activation of GABAA receptors eliciting anxiety-like behavior. Taken together these findings have helped clarify a neurochemical pathway controlling anxiety-like behavior evoked by acute stress in zebrafish while also opening the possibility of new perspective opportunities to use zebrafish as an animal model to test anxyolitic drugs that target the GABAergic system.

The GATs are the membrane proteins responsible for the uptake of GABA in the central nervous system. Alterations in GAT activity are implicated in several neurological diseases, including retinopathies. The present study describes an alternative method to determine GAT activity in tissue preparations of the central nervous system, using high performance liquid chromatography (HPLC) with fluorescence detection. The GABA concentration in the medium was determined using the o-phthaldehyde (OPA)-derivation protocol validated by the Brazilian Health Regulatory Agency (ANVISA) and the United States Food and Drug Administration (US-FDA). The GAT activity in the retinal preparations was determined through the evaluation of the GABA uptake, which was measured by assessing the difference between the initial and final concentrations of GABA in the incubation medium. The evaluation of the GAT kinetics returned values of K

Serotonin (5-HT) receptors have been implicated in responses to aversive stimuli in mammals and fish, but its precise role is still unknown. Moreover, since at least seven families of 5-HT receptors exist in vertebrates, the role of specific receptors is still debated. Aversive stimuli can be classified as indicators of proximal, distal, or potential threat, initiating responses that are appropriate for each of these threat levels. Responses to potential threat usually involve cautious exploration and increased alertness, while responses to distal and proximal threat involve a fight-flight-freeze reaction. We exposed adult zebrafish to a conspecific alarm substance (CAS) and observed behavior during (distal threat) and after (potential threat) exposure, and treated with the 5-HT2C receptor agonists MK-212 or WAY-161503 or with the antagonist RS-102221. The agonists blocked CAS-elicited defensive behavior (distal threat), but not post-exposure increases in defensive behavior (potential threat), suggesting inhibition of responses to distal threat. MK-212 blocked changes in freezing elicited by acute restraint stress, a model of proximal threat, while RS-102221 blocked changes in geotaxis elicited this stressor. We also found that RS-102221, a 5-HT2C receptor antagonist, produced small effect on behavior during and after exposure to CAS.Preprint:https://www.biorxiv.org/content/10.1101/2020.10.04.324202; Data and scripts:https://github.com/lanec-unifesspa/5-HT-CAS/tree/master/data/5HT2C

Glutamate release in response to a hypertonic stimulus is a well described phenomenon in the hypothalamus. Evidence suggests that hypothalamic glial cells release glutamate into the extracellular environment in hypertonic conditions. In the current study, we described autocrine regulation of adenosine on glutamate release induced by Na+hypertonicity in hypothalamic glial cell cultures. We showed that glial cells cultured from the cerebral cortex did not release glutamate or adenosine under hypertonic conditions. The findings suggest that the hypothalamus has specialized glial cells, which are responsive to osmotic variations. Stimulation or inhibition of adenosine A1 receptors modulates extracellular glutamate levels in hypothalamic glial cell cultures under hypertonic stimulation. Our results extend previous observations regarding the role of glial cells in the control of hypothalamic physiology. They further demonstrate for the first time that hypothalamic glial cells regulate Na+-hypertonicity-induced glutamate release by activation of adenosine A1 receptors via adenosine release.

Ototoxicity is a collateral effect of prolonged treatment with chloroquine which is a widely utilized as an anti-lupus and anti-malarial drug. Glial cells of inner ear are responsible for maintenance of neuronal cells homeostasis in auditory system. In the current study we have evaluated chloroquine-induced toxicity and protective effect of ascorbic acid treatment on Schwann glial cell cultures of inner ear. Glial cells were cultured from organ of Corti of mice cochlear structure. Purity of Schwann glial cell was confirmed by S100 protein staining. Cell viability was evaluated in control and cultures treated with different concentrations of chloroquine. Glutamate uptake and ROS production were measured by HPLC and DCFH-DA probe fluorescence, respectively. Results have shown that chloroquine treatment evoked concentration and time -dependent toxicity (LC50 = 70 μM) as well as significant decrease on glutamate uptake and high production of ROS in glial cell cultures. Co-treatment with ascorbic acid has prevented both chloroquine-induced ROS production and chloroquine toxicity on glial cell cultures. This pre-clinical study is the first one to demonstrate chloroquine-induced ROS production by glial cells of inner ear as well as the protective effect exerted by ascorbic acid on these cells.

Chronic exposure to mercury chloride (HgCl2) has been shown to promote oxidative stress and cell death in the central nervous system of adult rats displaying motor and cognitive impairments. However, there are no investigations about neurochemical function after this type of exposure in rodents that may be associated with those behavioral changes already reported. Thus, the aim of this study was to analyze glutamatergic and GABAergic dysfunctions in the motor cortex and hippocampus of adult rats, in a model of chronic exposure to HgCl2 in. Twenty rats were exposed to a daily dose of 0.375 mg/kg for 45 days. After this period, they were submitted to motor and cognitive functions tests and euthanized to collect the motor cortex and hippocampus for measurement of mercury (Hg) levels in the parenchyma and neurochemical assays for analysis of glutamatergic and GABAergic functions. It was observed that chronic exposure to HgCl2 promoted increase in total Hg levels in these two brain areas, with changes in glutamatergic transport, but without changes in GABAergic transport. Functionally this model of exposure caused the decrease of the spontaneous motor locomotion and in the process of learning and memory. In this way, our results provide evidences that glutamatergic neurochemical dysfunction can be pointed out as a strong causal factor of motor and cognitive deficits observed in rats exposed to this HgCl2.

Malaria is a life-threatening disease caused by Plasmodium and represents one of the main public health problems in the world. Among alterations associated with the disease, we highlight the hepatic impairment resulting from the generation of oxidative stress. Studies demonstrate that liver injuries caused by Plasmodium infection are associated with unbalance of the antioxidant system in hepatocytes, although little is known about the role of antioxidant molecules such as glutathione and vitamin C in the evolution of the disease and in the liver injury. To evaluate disease complications, murine models emerge as a valuable tool due to their similarities between the infectious species for human and mice. Herein, the aim of this study is to evaluate the effect of antioxidants glutathione and vitamin C on the evolution of murine malaria and in the liver damage caused by Plasmodium berghei ANKA infection. Mice were inoculated with parasitized erythrocytes and treated with glutathione and vitamin C, separately, both at 8 mg/kg during 7 consecutive days. Our data showed that during Plasmodium infection, treatment with glutathione promoted significant decrease in the survival of infected mice, accelerating the disease severity. However, treatment with vitamin C promoted an improvement in the clinical outcomes and prolonged the survival curve of infected animals. We also showed that glutathione promoted increase in the parasitemia rate of Plasmodium-infected animals, although treatment with vitamin C has induced significant decrease in parasitemia rates. Furthermore, histological analysis and enzyme biochemical measurement showed that treatment with glutathione exacerbates liver damage while treatment with vitamin C mitigates the hepatic injury induced by the infection. In summary, the current study provided evidences that antioxidant molecules could differently modulate the outcome of malaria disease; while glutathione aggravated the disease outcome and liver injury, the treatment with vitamin C protects the liver from damage and the evolution of the condition.

Rupture of Achilles tendon is a common accident affecting professional and recreational athletes. Acute and chronic pains are symptoms commonly observed in ruptured patients. Despite that, no studies have described whether Achilles tendon rupture is able to promote disorders in CNS. Based in these finds, the current study aimed to evaluate nociceptive alterations and inflammatory response in L5 lumbar segment of Balb/c mice spinal cord after Achilles tendon rupture. We demonstrated increased algesic response in the paw of ruptured group on the 7th and 14th days post tenotomy when compared with control group. This phenomenon was accompanied by over expression of COX-2 and NOS-2 as well as hyperactivation of astrocytes and microglia in nociceptive areas of L5 spinal cord as evidenced by intense GFAP and IBA-1 immunostaining, respectively. Biochemical studies also demonstrated increased levels of nitrite in the L5 spinal cord of tenotomized animals when compared with control group. Thus, we have demonstrate for the first time that total rupture of the Achilles tendon induces inflammatory response, nitrergic and glial activation in the CNS at L5 spinal cord region.

Astrocyte reactivity in the spinal cord may occur after peripheral neural damage. However, there is no data to report such reactivity after Achilles tendon injury. We investigate whether changes occur in the spinal cord, mechanical sensitivity and gait in two phases of repair after Achilles tendon injury. Wistar rats were divided into groups: control (CTRL, without rupture), 2 days post-injury (RUP2) and 21 days post-injury (RUP21). Functional and mechanical sensitivity tests were performed at 2 and 21 days post-injury (dpi). The spinal cords were processed, cryosectioned and activated astrocytes were immunostained by GFAP at 21 dpi. Astrocyte reactivity was observed in the L5 segment of the spinal cord with predominance in the white matter regions and decrease in the mechanical threshold of the ipsilateral paw only in RUP2. However, there was gait impairment in both RUP2 and RUP21. We conclude that during the acute phase of Achilles tendon repairment, there was astrocyte reactivity in the spinal cord and impairment of mechanical sensitivity and gait, whereas in the chronic phase only gait remains compromised., Achilles tendon, Tendon injury, Astrocyte reactivity, Gait, Mechanical sensitivity.

Sesamol is a phenolic derivative. Its antioxidant activity is low than that of Trolox and depends on benzodioxole moiety. Thus, a molecular modification strategy through alkylation, inspired by natural and synthetic antioxidants, was studied by molecular modeling at the DFT/B3LYP level of theory by comparing the 6-31+G(d,p) and 6-311++G(2d,2p) basis sets. All proposed derivatives were compared to classical related antioxidants such as Trolox, t-butylated hydroxytoluene (BHT) and t-butylated hydroxyanisole (BHA). According to our results, molecular orbitals, single electron or hydrogen-atom transfers, spin density distributions, and alkyl substitutions at the ortho positions related to phenol moiety were found to be more effective than any other positions. The trimethylated derivative was more potent than Trolox. t-Butylated derivatives were stronger than all other alkylated derivatives and may be new alternative forms of modified antioxidants from natural products with applications in the chemical, pharmaceutical, and food industries., PROPESP/UFPA, National Council for Scientific and Technological Development (CNPq)

Edaravone is a heterocyclic pyrazolone compound. It has pronounced effect against free radicals, however renal and hepatic disorders have been reported. Isoxazolones are considered bioisosteric analogues of pyrazolones and may have comparable properties. Thus, we investigated the structural and electronic influences for edaravone, isoxazolone, and their tautomers on antioxidant process. Structure and tautomerism study among edaravone, isoxazolone and their heterocycles derivatives were related to antioxidant mechanisms by using the hybrid DFT method B3LYP with the basis sets 6-31++G(2d,2p). The C-H tautomer was the most stable and energetically favored among them. Intramolecular N-H-N hydrogen bonds and polar medium were responsible for the low energy differences among all possible tautomers. N-H tautomers in both systems proved to be better antioxidant by SET (single electron transfer), while O-H tautomers were better antioxidant on HAT (homolytic hydrogen atom transfer) mechanism. Theoretical calculation showed that edaravone is more potent than phenylisoxazolone, however, both has similar antioxidant scavenging on experimental DPPH. The carbonyliminic system played a very important role in the antioxidant activity for both studied classes.

Current theories on the role of serotonin (5-HT) in vertebrate defensive behavior suggest that this monoamine increases anxiety but decreases fear, by acting at different levels of the neuroaxis. This paradoxical, dual role of 5-HT suggests that a serotonergic tone inhibits fear responses, while an acute increase in 5-HT would produce anxiety-like behavior. However, so far no evidence for a serotonergic tone has been found. Using zebrafish alarm responses, we investigate the participation of phasic and tonic 5-HT levels in fear-like behavior, as well as in behavior after stimulation. Conspecific alarm substance (CAS) increased bottom-dwelling and erratic swimming, and animals transferred to a novel environment after CAS exposure (post-exposure behavior) showed increased bottom-dwelling and freezing. Clonazepam blocked CAS effects during and after exposure. Acute fluoxetine dose-dependently decreased fear-like behavior, but increased post-exposure freezing. Metergoline had no effect on fear-like behavior, but blocked the effects of CAS on post-exposure behavior; similar effects were observed with pCPA. Finally, CAS was shown to decrease the activity of monoamine oxidase in the zebrafish brain after exposure. These results suggest that phasic and tonic serotonin encode an aversive expectation value, switching behavior towards cautious exploration/risk assessment/anxiety when the aversive stimulus is no longer present.

Panic disorder (PD) is characterized by recurrent and uncontrollable panic attacks associated with behavioral changes and/or persistent anxiety due to the attacks. The development of behavioral models in animals is important for the understanding of the psychobiological and behavioral bases of PD. The present article reviews the main models used in the current literature. Biobehavioral assays used in rats and mice include fear conditioning (which presents moderate predictive, face, and construct validities); the elevated T-maze (which presents good predictive validity, but low face and construct validities); electrical stimulation of the periaqueductal gray (which presents good face validity, but moderate construct validity); predator exposure models (which present good predictive and moderate construct validity); and hypercapnia-induced responses (which present moderate construct validity). These three approaches seek coherence with theories on fear as a way to increase its translational potential; thus, while the elevated T-maze is supported by the Deakin/Graeff theory, the mouse defense test battery relies on the concept of defensive distance, and periaqueductal gray stimulation is based on the functional neuroanatomy of fear. Moreover, to higher or lower degree the three models are supported by an “etho-experimental” approach, with careful observation of animal behavior as a way of discriminating different defensive strategies that model different aspects of anxiety, fear, and panic. These assays can be used, in conjunction with independent variables that attempt to simulate the vulnerabilities and stressors which lead to panic attacks, to produce true models of PD. Finally, an alternative/complementary model is proposed that uses zebrafish alarm reaction to study this disorder.

The cellular origins of slow ERG changes during light adaptation following a dark-adapted state are still unclear. To study light adaptation, six healthy, normal trichromats were dark-adapted for 30 min prior to full-field ERG recordings to sinusoidal stimuli that isolate responses of the L- or M-cones or that stimulate luminance and chromatic mechanisms at 12 or 36 Hz. Recordings were performed for 16 min with 2-min intervals after onset of a constant background. Generally, the responses were sine-wave-like, and the first harmonic (fundamental) component dominated the Fourier spectrum except for the 12-Hz luminance stimulus in which two components, a sine-wave-like component and a transient component, determined the response profiles, leading to large second harmonic components. The amplitude of the first harmonic component (F) increased as a function of the light-adaptation time except for the 12-Hz luminance stimulus at which the F component decreased as a function of the light-adaptation period. The phase of the first harmonic component changed only slightly (less than 30°) during the light-adaptation period for all stimuli conditions. The L/M ratio in luminance reflecting ERGs decreased with increasing adaptation time. Our present data suggest that the light-adaptation process mainly reflects changes in the luminance pathway. The responses to 12-Hz luminance stimuli are determined by two different luminance driven pathways with different adaptation characteristics.

Serotonin (5-HT) has been recognized as a neurotransmitter in the vertebrate retina, restricted mainly to amacrine and bipolar cells. It is involved with synaptic processing and possibly as a mitogenic factor. We confirm that chick retina amacrine and bipolar cells are, respectively, heavily and faintly immunolabeled for 5-HT. Amacrine serotonergic cells also co-express tyrosine hydroxylase (TH), a marker of dopaminergic cells in the retina. Previous reports demonstrated that serotonin transport can be modulated by neurotransmitter receptor activation. As 5-HT is diffusely released as a neuromodulator and co-localized with other transmitters, we evaluated if 5-HT uptake or release is modulated by several mediators in the avian retina. The role of different glutamate receptors on serotonin transport and release in vitro and in vivo was also studied. We show that l-glutamate induces an inhibitory effect on [3H]5-HT uptake and this effect was specific to kainate receptor activation. Kainate-induced decrease in [3H]5-HT uptake was blocked by CNQX, an AMPA/kainate receptor antagonist, but not by MK-801, a NMDA receptor antagonist. [3H]5-HT uptake was not observed in the presence of AMPA, thus suggesting that the decrease in serotonin uptake is mediated by kainate. 5-HT (10–50 μM) had no intrinsic activity in raising intracellular Ca2+, but addition of 10 μM 5-HT decreased Ca2+ shifts induced by KCl in retinal neurons. Moreover, kainate decreased the number of bipolar and amacrine cells labeled to serotonin in chick retina. In conclusion, our data suggest a highly selective effect of kainate receptors in the regulation of serotonin functions in the retinal cells.

Anxiety is a common symptom associated with high caffeine intake. Although the neurochemical mechanisms of caffeine-induced anxiety remain unclear, there are some evidences suggesting participation of oxidative stress. Based on these evidences, the current study is aimed at evaluating the possible protective effect of alpha-tocopherol (TPH) against anxiety-like behavior induced by caffeine (CAF) in zebrafish. Adult animals were treated with CAF (100 mg/kg) or TPH (1 mg/kg)+CAF before behavioral and biochemical evaluations. Oxidative stress in the zebrafish brain was evaluated by a lipid peroxidation assay, and anxiety-like behavior was monitored using light/dark preference and novel tank diving test. Caffeine treatment evoked significant elevation of brain MDA levels in the zebrafish brain, and TPH treatment prevented this increase. Caffeine treatment also induced anxiety-like behavior, while this effect was not observed in the TPH+CAF group. Taken together, the current study suggests that TPH treatment is able to inhibit oxidative stress and anxiety-like behavior evoked by caffeine.

Time-dependent sensitization (TDS)—the delayed increase in neurobehavioral responses to heterotypic stressors after exposure to an intense, inescapable stressor—has been proposed as an animal model for post-traumatic stress disorder (PTSD). Translationally relevant stressors used in TDS are capable of affecting more than one behavioral domain and produce interindividual variability in responsiveness. Here, conspecific alarm substance (CAS) is shown to induce TDS in zebrafish in inter- and intra-population-specific way. Exposure to CAS, an ecologically relevant stimulus which produces fear-like responses acutely, increased anxiety and arousal in zebrafish from the blue shortfin (BSF) phenotype 24 h after stimulus delivery. Anxiety-like responses were differently affected immediately and 24 h after stimulus delivery. Anxiety-like responses were more sensitized in zebrafish from the longfin (LOF) than in the BSF phenotype, an effect which is reminiscent of “basal” differences in anxiety-like behavior. After application of behavioral cutoff criteria, CAS was shown to produce intense TDS in ∼25% of LOF animals, while ∼20% of exposed animals showed little evidence of TDS. Overall, these results suggest that CAS induces TDS in zebrafish after a 24 h “incubation” period, with inter- and intra-population variability that underlines its face and ecological validity.

Glutamate release in response to a hypertonic stimulus is a well described phenomenon in the hypothalamus. Evidence suggests that hypothalamic glial cells release glutamate into the extracellular environment in hypertonic conditions. In the current study, we described autocrine regulation of adenosine on glutamate release induced by Na

Vertebrate retina has been shown to be an important target for mercury toxicity and very studies have shown the effect of mercury on the retinal ontogenesis. The nitrergic system plays an important role in the retinal development. The current work studied the effects of methylmercury (MeHg) exposure on the NO-synthase positive neurons (NADPH-diaphorase neurons or NADPH-d+) of the chick retinal ganglion cell layer at embryonic E15 and postnatal P1 days. Retinal flat mounts were stained for NADPH-diaphorase histochemistry and mosaic properties of NADPH-d + were studied by plotting isodensity maps and employing density recovery profile technique. It was also evaluated the protective effect of alpha-tocopherol treatment on retinal tissues exposed to MeHg. MeHg exposure decreased the density of NADPH-d + neurons and altered cell mosaic properties at E15 but had very little or no effect at P1 retinas. Alpha-tocopherol has a protective effect against MeHg exposure at E15. MeHg alterations and alpha-tocopherol protective effect in embryonic retinas were demonstrated to be at work in experimental conditions. MeHg effect in the early phases of visual system development in natural conditions might use the nitrergic pathway and supplementary diet could have a protective effect. At later stages, this mechanism seems to be naturally protected.

Chemical communication of predation risk has evolved multiple times in fish species, with conspecific alarm substance (CAS) being the most well understood mechanism. CAS is released after epithelial damage, usually when prey fish are captured by a predator and elicits neurobehavioural adjustments in conspecifics which increase the probability of avoiding predation. As such, CAS is a partial predator stimulus, eliciting risk assessment-like and avoidance behaviours and disrupting the predation sequence. The present paper reviews the distribution and putative composition of CAS in fish and presents a model for the neural processing of these structures by the olfactory and the brain aversive systems. Applications of CAS in the behavioural neurosciences and neuropharmacology are also presented, exploiting the potential of model fish [e.g., zebrafish Danio rerio, guppies Poecilia reticulata, minnows Phoxinus phoxinus) in neurobehavioural research.

The structure–antioxidant relationship of mangiferin was elucidated by means of quantum chemistry calculations. The density functional theory level of theory was used for antioxidant pharmacophore selection. According to the obtained geometry by using the DFT/B3LYP/6-31+G(d,p) values, HOMO, ionization potential, bond dissociation energy, stabilization energy, and spin density distribution, the catechol moiety is more important for the antioxidant capacity than the resorcinol moiety. However, the sugar is a strong electron-withdrawing group due to its hydrogen bond formation with the resorcinol moiety of xanthone ring. The hydrogen transfer is more important for the antioxidant activity than the electron transfer. The hydrogen abstractions in the ortho positions are more favored than the meta positions. Thus, our results show a synergistic effect between xanthone and sugar rings.

Background Cerebral malaria (CM) is a severe complication resulting from Plasmodium falciparum infection. This condition has usually been associated with cognitive, behavioural and motor dysfunctions, being the retinopathy the most serious consequence resulting from the disease. The pathophysiological mechanisms underlying this complication remain incompletely understood. Several experimental models of CM have already been developed in order to clarify those mechanisms related to this syndrome. In this context, the present work has been performed to investigate which possible electrophysiological and neurochemistry alterations could be involved in the CM pathology. Methods Experimental CM was induced in Plasmodium berghei-infected male and female C57Bl/6 mice. The survival and neurological symptoms of CM were registered. Brains and retina were assayed for TNF levels and NOS2 expression. Electroretinography measurements were recorded to assessed a- and b-wave amplitudes and neurochemicals changes were evaluated by determination of glutamate and glutathione levels by HPLC. Results Susceptible C57Bl/6 mice infected with ≈ 106 parasitized red blood cells (P. berghei ANKA strain), showed a low parasitaemia, with evident clinical signs as: respiratory failure, ataxia, hemiplegia, and coma followed by animal death. In parallel to the clinical characterization of CM, the retinal electrophysiological analysis showed an intense decrease of a- and-b-wave amplitude associated to cone photoreceptor response only at the 7 days post-infection. Neurochemical results demonstrated that the disease led to a decrease in the glutathione levels with 2 days post inoculation. It was also demonstrated that the increase in the glutathione levels during the infection was followed by the increase in the 3H-glutamate uptake rate (4 and 7 days post-infection), suggesting that CM condition causes an up-regulation of the transporters systems. Furthermore, these findings also highlighted that the electrophysiological and neurochemical alterations occurs in a manner independent on the establishment of an inflammatory response, once tumour necrosis factor levels and inducible nitric oxide synthase expression were altered only in the cerebral tissue but not in the retina. Conclusions In summary, these findings indicate for the first time that CM induces neurochemical and electrophysiological impairment in the mice retinal tissue, in a TNF-independent manner.

NMDA receptors have been implicated in the acute response to stress, possibly mediated the nitric oxide pathway; serotonin has also been implicated in these responses, and has recently been shown to modulate the nitric oxide pathway via 5-HT1 and 5-HT2 receptors. In this work, we compare the effects of NMDA and a 5-HT1A receptor ligands on light/dark preference in adult zebrafish, and investigate whether nitric oxide mediates the effects of such drugs. The noncompetitive NMDA receptor antagonist MK-801 decreased dark preference (scototaxis), while NMDA increased it; the effects of NMDA were completely blocked by pretreatment with the nitric oxide synthase (NOS) antagonist L-NAME. SNP, a nitric oxide donor, produced a bell-shaped dose–response profile on scototaxis. Treatment with 5-HTP increased scototaxis, an effect which was potentiated by pre-treatment with NMDA, but not MK-801, and partially blocked by L-NAME. The 5-HT1A receptor antagonist WAY 100,635 decreased scototaxis, an effect which was completely blocked by L-NAME. These results suggest that tonic NOS inhibition is an important downstream effector of 5-HT1A receptors in the regulation of dark preference behavior in zebrafish, and that NOS is also under phasic independent control by NMDA receptors.

Muscular atrophy is a progressive degeneration characterized by muscular proteolysis, loss of mass and decrease in fiber area. Tendon rupture induces muscular atrophy due to an intrinsic functional connection. Local inhibition of nitric oxide synthase (NOS) by No-nitro-L-arginine methyl ester (L-NAME) accelerates tendon histological recovery and induces functional improvement. Here we evaluate the effects of such local nitrergic inhibition on the pattern of soleus muscle regeneration after tenotomy. Adult male Wistar rats (240 to 280 g) were divided into four experimental groups: control (n=4), tenotomized (n=6), vehicle (n=6), and L-NAME (n=6). Muscular atrophy was induced by calcaneal tendon rupture in rats. Changes in muscle wet weight and total protein levels were determined by the Bradford method, and muscle fiber area and central core lesion (CCL) occurrence were evaluated by histochemical assays. Compared to tenotomized (69.3±22%) and vehicle groups (68.1%±17%), L-NAME treatment induced an increase in total protein level (108.3±21%) after 21 days post-injury. A reduction in fiber areas was observed in tenotomized (56.3±1.3%) and vehicle groups (53.9±3.9%). However, L-NAME treatment caused an increase in this parameter (69.3±1.6%). Such events were preceded by a remarkable reduction in the number of fibers with CCL in L-NAME-treated animals (12±2%), but not in tenotomized (21±2.5%) and vehicle groups (19.6±2.8%). Altogether, our data reveal that inhibition of tendon NOS contributed to the attenuation of atrophy and acceleration of muscle regeneration.

Muscular atrophy is a progressive degeneration characterized by muscular proteolysis, loss of mass and decrease in fiber area. Tendon rupture induces muscular atrophy due to an intrinsic functional connection. Local inhibition of nitric oxide synthase (NOS) by Nω-nitro-L-arginine methyl ester (L-NAME) accelerates tendon histological recovery and induces functional improvement. Here we evaluate the effects of such local nitrergic inhibition on the pattern of soleus muscle regeneration after tenotomy. Adult male Wistar rats (240 to 280 g) were divided into four experimental groups: control (n=4), tenotomized (n=6), vehicle (n=6), and L-NAME (n=6). Muscular atrophy was induced by calcaneal tendon rupture in rats. Changes in muscle wet weight and total protein levels were determined by the Bradford method, and muscle fiber area and central core lesion (CCL) occurrence were evaluated by histochemical assays. Compared to tenotomized (69.3±22%) and vehicle groups (68.1%±17%), L-NAME treatment induced an increase in total protein level (108.3±21%) after 21 days post-injury. A reduction in fiber areas was observed in tenotomized (56.3±1.3%) and vehicle groups (53.9±3.9%). However, L-NAME treatment caused an increase in this parameter (69.3±1.6%). Such events were preceded by a remarkable reduction in the number of fibers with CCL in L-NAME-treated animals (12±2%), but not in tenotomized (21±2.5%) and vehicle groups (19.6±2.8%). Altogether, our data reveal that inhibition of tendon NOS contributed to the attenuation of atrophy and acceleration of muscle regeneration.