Brain Ascorbic Acid Research Articles

Efficient Electrochemical Microsensor for the Simultaneous Measurement of Hydrogen Peroxide and Ascorbic Acid in Living Brains.

Hydrogen peroxide (H2O2) and ascorbic acid (AA), acting as two significant indicative species, correlate with the oxidative stress status in living brains, which have historically been considered to be involved mainly in neurodegenerative disorders such as Alzheimer's disease, Huntington's disease, and Parkinson's disease (PD). The development of efficient biosensors for the simultaneous measurement of their levels in living brains is vital to understand their roles played in the brain and their interactive relationship in the progress of these diseases. Herein, a robust ratiometric electrochemical microsensor was rationally designed to realize the determination of H2O2 and AA simultaneously. Therefore, a specific probe was designed and synthesized with both recognition units responsible for reacting with H2O2 to produce a detectable signal on the microsensor and linkage units helping the probe modify onto the carbon substrate. A topping ingredient, single-walled carbon nanotubes (SWCNTs) was added on the surface of the electrode, with the purpose of not only facilitating the oxidation of AA but also absorbing methylene blue (MB), prompting to read out the inner reference signal. This proposed electrochemical microsensor exhibited a robust ability to real-time track H2O2 and AA in linear ranges of 0.5-900 and 10-1000 μM with high selectivity and accuracy, respectively. Eventually, the efficient electrochemical microsensor was successfully applied to the simultaneous measurement of H2O2 and AA in the rat brain, followed by microinjection, and in the PD mouse brain.

An electrochemical microsensor based on a specific recognition element for the simultaneous detection of hydrogen peroxide and ascorbic acid in the live rat brain.

A novel electrochemical microsensor was developed for the ratiometric and simultaneous determination of hydrogen peroxide (H2O2) and ascorbic acid (AA) based on the borate-phenol "switch" recognition mechanism and carbon nanotube (CNT) catalytic characteristics. First of all, a carbon fiber microelectrode (CFME) was coated with CNTs. Then, a specific probe, 9-anthraceneboronic acid pinacol ester (9-AP), was screened and decorated on CNTs through π-π stacking for the recognition of H2O2 based on the transformation of boric acid ester into electroactive phenols. CNTs not only served as the amplifiers of current signals, but also as catalysts facilitating AA oxidation. Meanwhile, ferrocenecarboxylic acid (Fc), inert to H2O2 and AA, was modified on another amino-functionalized CNT microelectrode via an amide bond as an internal reference channel for avoiding errors caused by environmental discrepancies. The two-channel ratiometric microsensor enabled the sensitive and accurate detection of H2O2 and AA simultaneously, and the detection limits were estimated to be 0.09 μM and 4.12 μM, respectively. The developed microsensor with remarkable analytical performance was finally applied for the simultaneous detection of H2O2 and AA in the live rat brain.

Neuroprotective effect of and ascorbic acid in brain against high fat diet and stress induced neurotoxicity

Chronic exposure to stress and diet rich in saturated fat is one of the major reasons for the development of dementia and neurodegenerative disorders. The present study aims to examine the neuroprotective potential of and Ascorbic acid against high fat diet and stress induced neurotoxicity in brain. Animals were randomly divided into five groups. Group I received normal diet, Group II received high fat diet along with stress, Group III were treated with 100mg/kg body weight, and Group IV were treated with Ascorbic acid 100mg/kg body weight, Group V were treated with 100mg/kg body weight and Ascorbic acid 100mg/kg body weight. After the treatment all rats were sacrificed and brains were removed. Golgi staining was done and dendritic branching points and dendritic intersections were quantified with the help of cameralucida.There was a significant increase in dendritic length and branching points was observed in brain in rats treated with and Ascorbic acid.Present study concludes that and Ascorbic acid have neuroprotective role against high fat diet and stress induced Wistar rats.

Co-administration of nicotine ameliorates cannabis-induced behavioral deficits in normal rats: role of oxidative stress and inflammation

Nicotine (Nic) and cannabis are considered to be the most abused drugs worldwide that are progressively taken concomitantly. The present study aimed to investigate the modulatory effect of Nic on cannabis extract–induced neuro-inflammation, oxidative status, and the associated behavioral/biochemical alterations. Nic (0.25 mg/kg) and/or cannabis extract expressed as ∆9-tetrahydrocannabinol (THC10/20; 10 and 20 mg/kg) were given intraperitoneally for 30 days to Wistar rats. Nic shortened the floating time in forced swimming test, increased locomotion in the open field test, and decreased escape latency in the Morris water maze when co-administered with THC. These effects were associated with the inhibition of THC-mediated elevations in brain interleukin-1 beta, lipid peroxidation, superoxide dismutase, and ascorbic acid. Additionally, Nic increased serum butyrylcholinesterase (BChE) when combined with THC without affecting the serum acetylcholinesterase enzyme. The combinations spiked the brain glucose content above normal. In conclusion, the co-administration of Nic reduced THC-induced depressive-like behavior and memory impairment as well as hypo-locomotion associated with THC20. Such effects could be linked to Nic-mediated inhibition of brain oxidative stress, inflammation, and decreased serum BChE deactivity.

Galvanic Redox Potentiometry for Self-Driven in Vivo Measurement of Neurochemical Dynamics at Open-Circuit Potential.

Understanding the real-time correlation between chemical patterns and neural processes is critical for deciphering brain function. Voltammetry has enabled this task but with a number of challenges for current-based electrolysis in vivo. Herein, we report galvanic redox potentiometry (GRP) potentially as a universal strategy for in vivo monitoring of neurochemicals, with ascorbic acid (AA) as a typical example. The GRP sensor is constructed on a self-driven galvanic cell configuration, where AA is spontaneously oxidized at the indicating single-walled carbon nanotube-modified carbon fiber electrode (SWNT-CFE), while oxygen reduced at the laccase-modified reference CFE (Lac-CFE). At thermodynamic equilibrium, open-circuit potential (OCP) can be a linear indicator of the concentration of AA. The resulting sensor shows a high selectivity to AA dynamics in the presence of coexisting electroactive neurochemicals, which is mainly determined by the driving force for the cell reaction, as suggested by principal investigation. Sensing sensitivity of this OCP-based GRP method is not affected by nonspecific protein adsorption and electrode fouling. Moreover, a micropipette compartment of the reference electrode is designed to suppress mass crossover and prevent disturbance to oxygen reduction through confinement effect. The in vivo application of the GRP sensor is illustrated by measuring the basal level of cortical AA in live rat brain (230 ± 40 μM) and its dynamics during ischemia/reperfusion. The GRP concept is demonstrated as a prominent method for in vivo, real-time, quantitative analysis of brain neurochemistry.

Heterogeneous Nanostructure Design Based on the Epitaxial Growth of Spongy MoS x on 2D Co(OH)2 Nanoflakes for Triple-Enzyme Mimetic Activity: Experimental and Density Functional Theory Studies on the Dramatic Activation Mechanism.

In this study, we present a three-in-one catalytic platform for intrinsic oxidase-, peroxidase-, and catalase-like activity, which is enabled by epitaxial growth of the MoS x nanosponge on 2D Co(OH)2 nanoflakes [2D Co(OH)2 NFs] (CoMo hybrids). First, the 2D Co(OH)2 NFs are stripped from hierarchical three-dimensional Co(OH)2 nanoflowers which are synthesized in an eco-friendly way via one-step surfactant-free chemical route. Next, the porous MoS x nanosponge is decorated on the 2D Co(OH)2 NFs' surface using a solvothermal process forming heterogeneous nanostructured CoMo hybrids. Finally, because of the host-guest interaction, that is, after the epitaxial growth of spongy MoS x on 2D Co(OH)2 NFs, the heterogeneous nanostructure of CoMo hybrids exhibits unpredictable triple-enzyme mimetic activity simultaneously. The mechanisms of the oxidase-like properties are investigated by density functional theory (DFT) calculations, and it is discovered that a simple reaction/dissociation of O2 absorbed on Co-Mo thin films can explain the enhanced oxidase-like activity of the CoMo hybrids. In addition, the CoMo hybrids are also reproducible, stable, and reusable, that is, after 10 cycle uses, >90% mimic enzyme activity of the CoMo hybrids is still maintained. The oxidase-like activity of the CoMo hybrids enables it to oxidize 3,3',5,5'-tetramethylbenzidine (TMB) producing the blue oxTMB, which can selectively oxidize ascorbic acid (AA) and pave a new avenue for colorimetric sensing of AA. The proposed colorimetric strategy has been successfully utilized to measure AA in rat brain during the cerebral calm/ischemia process. Our findings provide in-depth insight into the future research of enzyme-like activities and might help to elucidate the mechanism and understand the role of epitaxial growth on the properties and application of hybrid nanostructures.

The CoOOH-TMB oxidative system for use in colorimetric and test strip based determination of ascorbic acid.

The authors report that cobalt oxyhydroxide (CoOOH) nanoflakes possess intrinsic oxidizing ability to directly oxidize 3,3',5,5'-tetramethylbenzidine (TMB) to form a blue colored product (oxTMB) even in the absence of H2O2 and oxygen. In the presence of ascorbic acid (AA), less of the blue product is formed because AA reduces oxTMB. These findings constitute a new scheme for colorimetric detection of AA. Absorbance, best measured at 652nm, linearly drops in the 10nM to 1μM AA concentration range, and the limit of detection is 5nM (at an S/N ratio of 3). The reaction is complete within <5s and highly selective. A strip test has been designed for fast and on-spot visual detection of AA. The method was applied to the direct estimation of AA in the microdialysate of brain, and also in soft drink samples. The strip test is considered to be a promising tool for the rapid screening of AA in brain and commercial samples. Graphic abstract Schematic of the CoOOH-TMB colorimetric system that exhibits a high selectivity for ascorbic acid (AA). A strip test has been designed for fast and on-spot visual detection of AA.

Anti‐Cancer Effects of Ascorbic Acid and Hyperbaric Oxygen Therapy in vitro

Cancer is the second leading cause of death in the U.S., projected to take 595,690 lives in 2016 and cost the nation over 125 billion dollars. To effectively reduce these detrimental losses, non‐toxic, low‐cost therapies should be further examined to supplement the standard of care. An anti‐carcinogenic, non‐toxic therapy currently under investigation is high dose ascorbic acid (AA). AA can function as a pro‐oxidant at pharmacological levels, delivering H2O2 to tumorous tissue upon oxidation and initiating cell death. At pharmacological concentrations (achieved i.v.), AA has shown significant anticancer effects in vitro, in vivo, and in small‐scale human reports at concentrations nontoxic to normal cells, thus having great potential as an adjuvant to the standard of care. Hyperbaric oxygen therapy (HBOT) is another non‐toxic, pro‐oxidant metabolic therapy that delivers 100% oxygen at elevated barometric pressure, elevating tissue pO2 and oxygenating hypoxic tumor cells, which, coupled with high levels of reactive oxygen and nitrogen species present in cancer cells, can further augment oxidative stress (OxS) and lead to cell death. This study's aims are as follows (1) to examine the anticancer effect of AA in vitro, (2) to evaluate the mechanism of AA‐induced OxS, (3) to investigate the potential synergy between AA and HBOT. We anticipate that this approach will yield significant insight into and further investigate the hypothesis that AA and HBOT can supplement the current standard of care.To characterize the anticancer effects of AA in vitro, we measured cell viability and proliferation following treatment with varying concentrations of AA in mouse brain tumor‐derived VM‐M3 cells. It was found that AA mediates cell death in a concentration‐dependent manner, with all tested concentrations ≥ 0.3mM AA significantly inducing cell death compared to control, specifically 0.3mM (p&lt;0.05) and 0.5– 5mM (p=0.0001). Preliminary results also indicate that concentrations ≥ 0.05mM AA inhibit cell proliferation compared to control and 0.01mM AA.To investigate the role of OxS in AA‐induced cytotoxicity, we measured VM‐M3 cell viability in the presence of AA and antioxidant N‐Acetylcysteine (NAC) and found that treatment with 5mM NAC attenuates the cytotoxic effect of AA (p&lt;0.0001). To evaluate the role of H2O2 in AA‐induced cytotoxicity, we will measure VM‐M3 cell viability in the presence of AA and the antioxidant enzyme catalase. Consistent with the literature, we anticipate that treatment with 300 units/mL of catalase will attenuate the cytotoxic effect of AA in vitro.To determine if HBOT can enhance the therapeutic effect of AA, we measured measure VM‐M3 cell viability following treatment with HBOT and AA. We found that HBOT significantly enhanced the cytotoxic effect of 0.3mM AA (p&lt;0.001). To complete this aim, we will measure VM‐M3 cell proliferation following treatment with HBOT, HBOT pre‐treatment, and AA.This data indicates that AA exhibits anti‐cancer effect in vitro through an OxS mechanism and that HBOT can enhance this therapeutic effect. These non‐toxic, pro‐oxidant metabolic therapies should be further investigated as adjuvants to the current standard of care.Support or Funding InformationWork supported by: USF Foundation (501c3); Metabolic Therapy and Cancer Research Account (#250244): Scivation: Florida High Tech Corridor Funding (#MED052‐0061361)

Dietary pyridoxine promotes growth and cellular metabolic plasticity of C hanos chanos fingerlings exposed to endosulfan induced stress

An attempt was made to assess the effect of dietary pyridoxine supplementation on growth performance and cellular metabolic enzymes of Chanos chanos fingerlings under endosulfan induced stress. Two-hundred and twenty-five fish were randomly distributed into five treatments, each with three replicates and fed four isocaloric and isonitrogenous diets with graded levels of pyridoxine for 45 days. Endosulfan concentration in the water was maintained at sublethal level of 1/40th of 96 h LC50 (0.52 ppb). Dietary pyridoxine supplementation significantly enhanced growth performance viz. weight gain (%), specific growth rate, feed conversion ratio and protein efficiency ratio of fish compared to control diet fed group. The results of this experiment revealed that the activities of digestive and metabolic enzymes were significantly improved after feeding with dietary pyridoxine supplementation in C. chanos. Significantly increased muscle and brain ascorbic acid content was observed in fish fed diets containing pyridoxine. Histological studies on liver of fish fed pyridoxine-supplemented diet showed the normal architecture of hepatocytes compared to non-pyridoxine fed group. The results obtained in this study revealed that the inclusion of pyridoxine in diet has a protective effect against endosulfan induced stress in C. chanos fingerlings through metabolic enzyme activities and vitamin C content. This study has a potential in ameliorating the toxic effect of endosulfan induced stress using dietary supplementation of pyridoxine.

Low brain ascorbic acid increases susceptibility to seizures in mouse models of decreased brain ascorbic acid transport and Alzheimer's disease

Seizures are a known co-occurring symptom of Alzheimer's disease, and they can accelerate cognitive and neuropathological dysfunction. Sub-optimal vitamin C (ascorbic acid) deficiency, that is low levels that do not lead the sufferer to present with clinical signs of scurvy (e.g. lethargy, hemorrhage, hyperkeratosis), are easily obtainable with insufficient dietary intake, and may contribute to the oxidative stress environment of both Alzheimer's disease and epilepsy. The purpose of this study was to test whether mice that have diminished brain ascorbic acid in addition to carrying human Alzheimer's disease mutations in the amyloid precursor protein (APP) and presenilin 1 (PSEN1) genes, had altered electrical activity in the brain (electroencephalography; EEG), and were more susceptible to pharmacologically induced seizures. Brain ascorbic acid was decreased in APP/PSEN1 mice by crossing them with sodium vitamin C transporter 2 (SVCT2) heterozygous knockout mice. These mice have an approximately 30% decrease in brain ascorbic acid due to lower levels of SVCT2 that supplies the brain with ASC. SVCT2+/-APP/PSEN1 mice had decreased ascorbic acid and increased oxidative stress in brain, increased mortality, faster seizure onset latency following treatment with kainic acid (10 mg/kg i.p.), and more ictal events following pentylenetetrazol (50 mg/kg i.p.) treatment. Furthermore, we report the entirely novel phenomenon that ascorbic acid deficiency alone increased the severity of kainic acid- and pentylenetetrazol-induced seizures. These data suggest that avoiding ascorbic acid deficiency may be particularly important in populations at increased risk for epilepsy and seizures, such as Alzheimer's disease.

The optimal dietary level of ascorbic acid for juvenile Pacific bluefin tuna, Thunnus orientalis

A feeding experiment was conducted to evaluate the efficacy of, and determine the requirements for dietary AsA (l-ascorbic acid) using its popular derivative l-ascorbyl-2-monophosphate-Mg (APM) for juvenile Pacific bluefin tuna (PBT). The five test diets (with APM-0, -400, -800, -1200 and -1600) were prepared by adding 0, 400, 800, 1200 and 1600 mg of APM kg diet−1, respectively. These diets, together with chopped sand lance (the control diet), were administered, in duplicate to 0.27 g PBT at 25 days after hatching (300 juveniles per 15 m3 tank). The PBT were fed to apparent satiety 6 times a day for 2 weeks. While final body weight, feed conversion and survival showed no differences among diets APM 400–1600, PBT fed APM-0 indicated lower growth performance together with anorexia, dark pigmentation and ataxia from 5 days and reached 80 % mortality at 12 days after the start of the feeding trial. In addition, liver and brain ascorbic acid (AsA) concentrations increased with increasing dietary APM levels, the values were similar in PBT fed diets with APM-1200 and APM-1600, and liver and brain concentrations of AsA were significantly higher in these fish than in the PBT fed the other diets (P < 0.05). The PBT responded to increasing dietary APM, and the fish fed APM-1200 had the highest weight gain, protein and lipid content. The data indicate that PBT can utilize APM as an AsA source and suggest that dietary supplementation with APM-1200 (454 mg AsA kg−1 diet) would result in satisfactory growth.

Antioxidant activity and low toxicity of (E)-1-(1-(methylthio)-1-(selenopheny) hept-1-en-2-yl) pyrrolidin-2-one

The aim of the present study was to evaluate the potential pharmacological and toxicological properties of (E)-1-(1-(methylthio)-1-(selenopheny) hept-1-en-2-yl) pyrrolidin-2-one (compound 1), an organoselenium compound. In vitro experiments showed that compound 1 presented a reduction in the lipid peroxidation induced by Fe²⁺ in thiobarbituric acid-reactive species (TBARS) production, and in the generation of reactive species caused by Fe²⁺/malonate in DCFH-DA oxidation. The high dose (500 mg/kg) induced an increase on ALT but not on AST activity. Hepatic, but not cerebral, δ-ALA-D activity from mice treated with 500 mg/kg presented a significant inhibition. Brain catalase activity was significantly inhibited by 100 mg/kg whereas hepatic catalase activity showed a significant increase at all doses. Hepatic lipid peroxidation was diminished only at lowest dose (100 mg/kg) whereas for brain tissue, all doses induced a significant reduction in TBARS levels. Brain and liver ascorbic acid contents were increased only at highest dose of compound 1. Urea and creatinine levels were not significantly altered by treatments. This is a promising compound with antioxidant activity and low toxicity, suggesting the potential beneficial activity of compound 1 against oxidative damage in many parameters studied in rats and mice.

Disruption of non-enzymatic antioxidant defense systems in the brain of rats with water-immersion restraint stress

We examined whether non-enzymatic antioxidant defense systems are disrupted in the brain of rats with water-immersion restraint stress. When rats were exposed to water-immersion restraint stress for 1.5, 3 or 6 h, the brain had decreased ascorbic acid and reduced glutathione contents and increased lipid peroxide and nitric oxide metabolites contents at 3 h and showed further changes in these components with a reduction of vitamin E content at 6 h. Increased serum levels of stress markers were found at 1.5, 3 or 6 h of WIRS. Oral pre-administration of L-ascorbic acid (1.5 mmol/kg) or vitamin E (0.5 mmol/kg) to rats with 6 h of water-immersion restraint stress attenuated the increases in lipid peroxide and nitric oxide metabolites contents and the decrease in vitamin E content in the brain. Pre-administered L-ascorbic acid attenuated the decreases in brain ascorbic acid and reduced glutathione contents at 6 h of water-immersion restraint stress, while pre-administered vitamin E enhanced the decreases in those contents. Pre-administered L-ascorbic acid or vitamin E did not affect the increased serum levels of stress markers in rats with 6 h of water-immersion restraint stress. These results indicate that water-immersion restraint stress causes disruption of non-enzymatic antioxidant defense systems through enhanced lipid peroxidation and nitric oxide generation in the brain of rats with water-immersion restraint stress.

Level of L-ascorbyl-2-monophosphate-Mg as a Vitamin C Source in Practical Diets for the Asian sea bass, Lates calcarifer

The stable vitamin C derivative, L-ascorbyl-2-monophosphate-Mg (AMP), was used as a supplement in practical diets for Asian sea bass, Lates calcarifer. Optimum growth, liver ascorbic acid (AA) saturation, and wound healing were determined. Sea bass (78.9±0.4 g) were fed a maintenance diet without vitamin C supplement for 25 days, then distributed into fifteen 1500-l oval fiberglass tanks at 30 fish each and fed one of five practical diets containing 0, 50, 100, 200, or 400 AMP mg/kg diet for 14 weeks. Fish fed the AMP-free diet exhibited clinical signs of vitamin C deficiency and significantly poorer final average weight, specific growth rate, protein efficiency ratio, feed conversion ratio, and hematocrit level (p&lt;0.05). The level of AA in the sea bass brain increased as the level of dietary AMP increased. The AA concentration in the liver was similar at all AMP dietary levels. Body calcium of sea bass fed the AMP-free or 50 mg diets was significantly lower (p&lt;0.05) than in fish fed the other diets. AMP enhanced wound healing regardless of supplement level, but fish fed the highest dietary AMP (400 mg/kg diet) exhibited histopathological changes in the liver. The dietary level of 50 mg AMP/kg diet was adequate for optimum growth, liver AA saturation, and prevention of clinical signs of vitamin C deficiency in 80-220 g sea bass. However, 100 mg AMP per kg diet was necessary for optimum body calcium.

Cerebral monoamine metabolism in guinea-pigs with ascorbic acid deficiency.

Guinea-pigs kept on a diet deficient in vitamin C showed, after 3 weeks, a marked decrease of ascorbic acid in brain and blood leucocytes as well as of the activity of alkaline phosphatase in blood plasma. Pair-fed animals did not exhibit these changes. The alpha-methyl-p-tyrosine (alpha MpT)-induced diminution of noradrenaline in the hypothalamus and the rest of the brain was attenuated in pair-fed animals, but restored in guinea-pigs deficient in ascorbic acid. The cerebral noradrenaline content (without administration of alpha MpT) showed a decrease in both pair-fed and ascorbic acid deficient animals. The noradrenaline of the heart exhibited a similar tendency. The alpha MpT-induced dopamine decrease in the striatum of ascorbic acid deficient animals was attenuated and the dopamine content (without alpha MpT administration) decreased. Pair-fed animals showed a similar tendency. The striatal concentration of homovanillic acid (HVA) was diminished in both pair-fed and ascorbic acid deficient guinea-pigs. The cerebral content of 5-hydroxyindoleacetic acid showed a decrease in pair-fed as well as in ascorbic acid deficient animals. It is concluded that ascorbic acid deficiency enhances the turnover of brain noradrenaline, whereas under-nutrition without ascorbic acid deficiency (pair-feeding) diminishes the turnover of cerebral noradrenaline, 5-hydroxytryptamine and striatal dopamine.

Growth and Feed Efficiency in Mangrove Red Snapper, (Lutjanus argentimaculatus Forsskal 1775) Fed Practical Diets Supplemented with L-ascorbyl-2-monophosphate-Mg

Growth and feed efficiency were determined in red snapper, Lutjanus argentimaculatus (Forsskal 1775), fed diets containing L-ascorbyl-2-monophosphate-Mg (AMP). Fish (13.39±0.08 g) were fed a practical diet without vitamin C supplement for four weeks then stocked in twelve 650-l tanks at 30 fish/tank and fed one of four practical diets containing AMP at 0, 60, 180, or 540 mg/kg dry diet for 17 weeks. Survival rates in all treatments were similar (88.9-98.9%). Fish fed the 0 or 540 ppm diets had inferior final average weights, protein efficiency ratios, and feed conversion ratios than fish fed the 60 or 80 ppm diets (p&lt;0.05). Growth of fish fed the 0 or 540 ppm diets slowed down on day 60 and fish fed the AMP-free diet exhibited clinical signs of vitamin C deficiency with a soft body and a significantly high (p&lt;0.05) hepatosomatic index. Ascorbic acid in brain and liver tissues rose with the level of dietary AMP. Fish fed the 540 ppm diet had significantly lower hematocrit (p&lt;0.05) than fish fed the 60 or 180 ppm diets. Histological analysis of the liver and kidney of fish fed the 180 and 540 ppm diets showed changes indicative of possible toxic effects. Based on growth, feed efficiency, tissue histology, and hematocrit level, AMP at 540 ppm is toxic to snapper. Thus, supplementation of 60 ppm AMP or its equivalent 26 ppm ascorbic acid in practical diets for red snapper promotes optimum growth and feed efficiency and prevents vitamin C deficiency symptoms.

Effect of a polyphenol-rich wild blueberry extract on cognitive performance of mice, brain antioxidant markers and acetylcholinesterase activity

The aim of this study was to examine the effect of a polyphenol-rich extract (PrB) of Vaccinium angustifolium (wild blueberries) introduced intraperitonealy (i.p.) at 30 (PrB30) and 60 (PrB60) mg/kg body weight for 7 days, on cognitive performance, brain oxidative status and acetylcholinesterase activity in adult, male, 3–4-month-old Balb-c mice. Evaluation of rodent learning and memory was assessed by a step-through test on day 6 after a double training and an initial acquisition trial on day 5. Antioxidant status was determined by ferric reducing antioxidant power (FRAP), ascorbic acid concentration (FRASC), malondialdehyde and reduced glutathione levels in whole brain homogenates. Acetylcholinesterase (AChE) activity was determined by Ellman's colorimetric method. Results showed that the PrB60-treated mice exhibited a significant improvement in learning and memory (step-through latency time of 228 ± 38 s compared to 101 ± 32 s of the control group). PrB extract administration also resulted in reduced lipid peroxidation products (38 and 79%) and higher brain ascorbic acid levels (21 and 64%) in both PrB30 and PrB60-treated groups, respectively, and higher glutathione levels (28%) in the PrB60-treated group. Furthermore, salt- and detergent soluble AChE activity significantly decreased in both PrB-treated groups. Thus, the significant cognitive enhancement observed in adult mice after short-term i.p. supplementation with the blueberry extract concentrated in polyphenols, is closely related to higher brain antioxidant properties and inhibition of AChE activity. These findings stress the critical impact of wild blueberry bioactive components on brain function.

Carbon Nanotube-Modified Carbon Fiber Microelectrodes for In Vivo Voltammetric Measurement of Ascorbic Acid in Rat Brain

This study demonstrates a new electrochemical method for in vivo measurements of ascorbic acid (AA) in rat brain with multiwalled carbon nanotube (MWNT)-modified carbon fiber microelectrodes (CFMEs) based on the electrochemical property of MWNTs for facilitating AA oxidation. Cyclic voltammetry results indicate that the prepared MWNT-modified CFMEs possess a marked electrocatalytic activity toward AA oxidation and can be used for its selective measurement in the presence of other kinds of electroactive species coexisting in rat brain, such as 3,4-dihydroxyphenylacetic acid, uric acid, and 5-hydroxytryptamine. The selectivity of the MWNT-modified CFMEs toward AA measurement is further studied in vivo by exogenously infusing ascorbate oxidase into the brain, and the results confirm that the prepared electrodes are selective and can thus be used for reliable in vivo measurements of AA in rat brain, combined with their good stability during in vivo measurements. The basal level of striatum AA is determined to be 0.20 +/- 0.05 mM (n = 3). The application of the voltammetric method with the MWNT-modified CFMEs is preliminarily demonstrated for in vivo observation of homeostatic regulation of striatum AA with exogenous infusion of AA into the brain.

Neuroprotective effect of Azadirachta indica on cerebral post-ischemic reperfusion and hypoperfusion in rats

We assessed the effect of Azadirachta indica ( A. indica), a plant that has been reported to possess antioxidant, anti-inflammatory and anxiolytic properties, on cerebral reperfusion injury and long term cerebral hypoperfusion. When blood flow to brain region that has undergone critical period of ischemia is re-established, additional injury is to be expected from the reperfusion. In the present study, bilateral common carotid artery (BCCA) occlusion for 30 min followed by 45 min reperfusion resulted in increase in lipid peroxidation, superoxide dismutase (SOD) activity and fall in total tissue sulfhydryl (T-SH) groups. A. indica pretreatment (500 mg/kg/day × 7 days) attenuated the reperfusion induced enhanced lipid peroxidation, SOD activity and prevented fall in T-SH groups. Moreover, A.indica per se increased brain ascorbic acid level, which was unchanged during reperfusion insult. Long-term cerebral hypoperfusion induced by permanent BCCA occlusion has been reported to cause behavioral and histopathological abnormalities. In the present study, as tested by open field paradigm and Morris' water maze, a propensity towards anxiety and disturbances of learning/memory were observed in animals subjected to hypoperfusion for 2 weeks. A. indica (500 mg/kg/day × 15 days) significantly reduced these hypoperfusion induced functional disturbances. Reactive changes in brain histology like gliosis, perivascular lymphocytic infiltration, recruitment of macrophages and cellular edema following long term hypoperfusion were also attenuated effectively by A. indica. We conclude that our study provides an experimental evidence for possible neuroprotective potentiality of A. indica.

Oxidative stress following traumatic brain injury in rats

Free radicals may be involved in the pathophysiology of traumatic brain injury (TBI) through oxidative damage of neurovascular structures. Endogenous antioxidants, such as ascorbate and alpha-tocopherol, may play a critical role in combating these oxidative reactions and their oxidized products can serve as an important index of oxidative stress. We used electron spin resonance (ESR) spectroscopy and in vivo spin trapping (reaction of an organic compound with free radical species) to detect the possible generation of free radicals after TBI. Injury was inflicted by a weight drop technique over the head (5.7 kg-cm). Rats were intravenously infused with either 1 mL, 0.1 M of the spin trap, alpha-phenyl-N-tert-butyl nitrone (PBN), or an equivalent volume of saline immediately before TBI or sham-injury. Animals were divided into four groups: (1) Group I: PBN-infused sham-injured, (2) Group II: PBN-infused injured, (3) Group III: saline-infused sham-injured, and (4) Group IV: saline-infused injured. Additional groups of saline-infused uninjured, saline-infused, and PBN-infused injured animals were used for histopathology. Sixty minutes after TBI or sham-injury, rats were again anesthetized and decapitated. The brains were removed within 1 minute, homogenized, and extracted for lipids. The extracts were analyzed by ESR spectroscopy. Brain ascorbic acid (AA) concentration was determined spectrophotometrically, using the ascorbate oxidase assay. No PBN spin adduct signals (indicating trapped free radical species) were visible 60 minutes after TBI. All groups of rats showed an ascorbyl free radical signal. The ascorbyl signal intensity (AI) was, however, significantly higher in the injured rats, while the brain (AA) was significantly reduced. In addition, the ratio of AI/AA, which eliminates the effect of variable ascorbate concentrations in the brain, was also significantly higher in the injured animals. We conclude that 60 minutes following TBI there was a significantly increased level of oxidative stress in the brain. This may reflect formation of free radical species with subsequent interaction with ascorbate (antioxidant) during the 60 minute period. The lack of PBN spin adduct signals 1 hour after TBI may indicate that free radical generation is time dependent and might be detectable earlier or later than the 60 minute period.