Ascorbate Levels Research Articles

Effect of varying high temperatures during reproductive growth on reproductive function, oxidative stress and seed yield in chickpea genotypes differing in heat sensitivity

The mechanisms affecting the heat sensitivity of chickpea are largely unknown. Heat-tolerant (ICCV07110, ICCV92944) and heat-sensitive (ICC14183, ICC5912) chickpea genotypes were sown in February in the soil-filled pots. At the time of flowering, these were subjected to varying day/night temperatures of 30/20, 35/25, 40/30 and 45/35°C in the growth chambers (12 h light/12 h dark; light intensity, 250 μmol m−2 s−1, 80% relative humidity). The pollen viability, pollen germination, tube growth, pollen load and stigma receptivity decreased with increases in temperatures to 45/35°C. The heat-tolerant genotypes experienced significantly less damage to pollen and stigma function. Membrane integrity, chlorophyll content, photochemical efficiency and cellular oxidizing ability were inhibited by the increase in temperature, with greater impacts on the sensitive genotypes. Oxidative injury as lipid peroxidation and hydrogen peroxide content was significantly greater in sensitive genotypes at 40/30 and 45/35°C. Enzymatic and non-enzymatic antioxidants showed increased levels at 40/30°C, but decreased considerably at 45/35°C. Heat-tolerant genotypes possessed greater activity of ascorbate peroxidase and glutathione reductase, along with higher levels of ascorbate and reduced glutathione at 40/30 and 45/35°C. Biomass, pod set and yield were not affected significantly at 35/25°C, but began to decrease at 40/30°C and were lowest at 45/35°C. The sensitive genotypes were not able to set any pods at 45/35°C, whereas the tolerant genotypes produced only few fertile pods at this temperature. It was concluded that heat stress leads to loss of pollen as well as stigma function and induces oxidative stress in the leaves that cause failure of fertilization and damage to the leaves, respectively.

Clinically Insignificant Effect of Supplemental Vitamin C on Serum Urate in Patients With Gout: A Pilot Randomized Controlled Trial

Studies in human volunteers have shown that vitamin C reduces serum urate (SU) levels. The aim of this study was to determine the effects of vitamin C on SU levels in patients with gout. Patients with gout and an SU level >0.36 mmoles/liter (6 mg/dl) were recruited. Twenty patients already taking allopurinol were randomized to receive an increase in the dose of allopurinol or to commence taking vitamin C (500 mg/day). Twenty patients who had not been taking allopurinol were randomized to start receiving either allopurinol (up to 100 mg/day) or vitamin C (500 mg/day). Levels of plasma ascorbate, creatinine, and SU were measured on day 0 and week 8. There was no significant difference in the baseline SU level or estimated glomerular filtration rate (eGFR) between those who received vitamin C and those who did not (for SU, mean ± SEM 0.50 ± 0.11 mmoles/liter [8.4 ± 1.8 mg/dl] versus 0.50 ± 0.09 mmoles/liter [8.4 ± 1.5 mg/dl]; for eGFR, mean ± SEM 65.5 ± 3.5 ml/minute/1.73 m(2) versus 67.9 ± 4.6 ml/minute/1.73 m(2) ). Among the randomized patients, 30% in the vitamin C group and 25% in the no vitamin C control group were receiving diuretics. In the patients receiving vitamin C, there was a significant increase between day 0 and week 8 in the plasma ascorbate level. The reduction in SU level over 8 weeks was significantly less in those patients receiving vitamin C compared to those who started or increased the dose of allopurinol (mean reduction 0.014 mmoles/liter [0.23 mg/dl] versus 0.118 mmoles/liter [1.9 mg/dl]; P < 0.001). A modest dosage of vitamin C (500 mg/day) for 8 weeks had no clinically significant urate-lowering effects in patients with gout, despite the fact that plasma ascorbate levels increased. These results differ from previous findings in healthy control subjects with hyperuricemia. The uricosuric effect of modest-dose vitamin C appears to be small in patients with gout, when administered as monotherapy or in combination with allopurinol.

Insulin, Ascorbate, and Glucose Have a Much Greater Influence Than Transferrin and Selenous Acid on theIn VitroGrowth of Engineered Cartilage in Chondrogenic Media

The primary goal of this study was to characterize the response of chondrocyte-seeded agarose constructs to varying concentrations of several key nutrients in a chondrogenic medium, within the overall context of optimizing the key nutrients and the placement of nutrient channels for successful growth of cartilage tissue constructs large enough to be clinically relevant in the treatment of osteoarthritis (OA). To this end, chondrocyte-agarose constructs (ø4×2.34 mm, 30×10(6) cells/mL) were subjected to varying supplementation levels of insulin (0× to 30× relative to standard supplementation), transferrin (0× to 30×), selenous acid (0× to 10×), ascorbate (0× to 30×), and glucose (0× to 3×). The quality of resulting engineered tissue constructs was evaluated by their compressive modulus (E(-Y)), tensile modulus (E(+Y)), hydraulic permeability (k), and content of sulfated glycosaminoglycans (sGAG) and collagen (COL); DNA content was also quantified. Three control groups from two separate castings of constructs (1× concentrations of all medium constituents) were used. After 42 days of culture, values in each of these controls were, respectively, E(-Y)=518±78, 401±113, 236±67 kPa; E(+Y)=1420±430, 1140±490, 1240±280 kPa; k=2.3±0.8×10(-3), 5.4±7.0×10(-3), 3.3±1.3×10(-3) mm(4)/N·s; sGAG=7.8±0.3, 6.3±0.4, 4.1±0.5%/ww; COL=1.3±0.2, 1.1±0.3, 1.4±0.4%/ww; and DNA=11.5±2.2, 12.1±0.6, 5.2±2.8 μg/disk. The presence of insulin and ascorbate was essential, but their concentrations may drop as low as 0.3× without detrimental effects on any of the measured properties; excessive supplementation of ascorbate (up to 30×) was detrimental to E(-Y), and 30× insulin was detrimental to both E(+Y) and E(-Y). The presence of glucose was similarly essential, and matrix elaboration was significantly dependent on its concentration (p<10(-6)), with loss of functional properties, composition, and cellularity observed at ≤0.3×; excessive glucose supplementation (up to 3×) showed no detrimental effects. In contrast, transferrin and selenous acid had no influence on matrix elaboration. These findings suggest that adequate distributions of insulin, ascorbate, and glucose, but not necessarily of transferrin and selenous acid, must be ensured within large engineered cartilage constructs to produce a viable substitute for joint tissue lost due to OA.

Exposure to elevated ozone levels differentially affects the antioxidant capacity and the redox homeostasis of two subtropical Phaseolus vulgaris L. varieties

Ozone (O3) has become one of the most toxic air pollutants to plants worldwide. However, investigations on O3 impacts on crops health and productivity in South America countries are still scarce. The present study analyzed the differences on the enzymatic and non-enzymatic antioxidant system in foliar tissue of two subtropical Phaseolus vulgaris varieties exposed to high O3 concentration. Both varieties were negatively impacted by the pollutant, but the responses between each variety were quite distinct. Results revealed that Irai has higher constitutive levels of reactive oxygen species (ROS) and ascorbate (AsA) concentration, but lower total thiol levels and catalase immunocontent. In this variety catalase protein concentration was increased after O3 exposure, indicating a better cellular capacity to reduce hydrogen peroxide. On the opposite, Fepagro 26-exposed plants increased ROS generation and AsA concentration, but had the levels of total thiol content and catalase protein unchanged. Furthermore, O3 treatment reduced the levels of chlorophylls a and b, and the relationship analysis between the chlorophyll ratio (a/b) and protein concentration were positively correlated indicating that photosynthetic apparatus is compromised, and thus probably is the biomass acquisition on Fepagro 26. Differently, O3 treatment of Irai did not affect chlorophylls a and b content, and loss on the protein content was lower. Altogether, these data suggest that early accumulation of ROS on Fepagro 26 are associated with an insufficient leaf antioxidant capacity, which leads to cell structure disruption and impairs the photosynthesis. Irai seems to be more tolerant to O3 toxic effects than Fepagro 26, and the observed differences on O3 sensitivity between the two varieties are apparently based on constitutive differences involved in the maintenance of intracellular redox homeostasis.

Effect of Cerium Oxide Nanoparticles on Rice: A Study Involving the Antioxidant Defense System and In Vivo Fluorescence Imaging

Previous studies have reported the uptake of cerium oxide nanoparticles (nCeO2) by plants, but their physiological impacts are not yet well understood. This research was aimed to study the impact of nCeO2 on the oxidative stress and antioxidant defense system in germinating rice seeds. The seeds were germinated for 10 days in nCeO2 suspension at 62.5, 125, 250, and 500 mg L(-1) concentrations. The Ce uptake, growth performance, stress levels, membrane damage, and antioxidant responses in seedlings were analyzed. Ce in tissues increased with increased nCeO2 concentrations, but the seedlings showed no visible signs of toxicity. Biochemical assays and in vivo imaging of H2O2 revealed that, relative to the control, the 62.5 and 125 mg nCeO2 L(-1) treatments significantly reduced the H2O2 generation in both shoots and roots. Enhanced electrolyte leakage and lipid peroxidation were found in the shoots of seedlings grown at 500 mg nCeO2 L(-1). Altered enzyme activities and levels of ascorbate and free thiols resulting in enhanced membrane damage and photosynthetic stress in the shoots were observed at 500 mg nCeO2 L(-1). These findings demonstrate a nCeO2 concentration-dependent modification of oxidative stress and antioxidant defense system in rice seedlings.

RNAi mediated down regulation of myo-inositol-3-phosphate synthase to generate low phytate rice

BackgroundPhytic acid (InsP6) is considered as the major source of phosphorus and inositol phosphates in cereal grains. Reduction of phytic acid level in cereal grains is desirable in view of its antinutrient properties to maximize mineral bioavailability and minimize the load of phosphorus waste management. We report here RNAi mediated seed-specific silencing of myo-inositol-3-phosphate synthase (MIPS) gene catalyzing the first step of phytic acid biosynthesis in rice. Moreover, we also studied the possible implications of MIPS silencing on myo-inositol and related metabolism, since, first step of phytic acid biosynthesis is also the rate limiting step of myo-inositol synthesis, catalyzed by MIPS.ResultsThe resulting transgenic rice plants (T3) showed a 4.59 fold down regulation in MIPS gene expression, which corresponds to a significant decrease in phytate levels and a simultaneous increment in the amount of inorganic phosphate in the seeds. A diminution in the myo-inositol content of transgenic plants was also observed due to disruption of the first step of phytic acid biosynthetic pathway, which further reduced the level of ascorbate and altered abscisic acid (ABA) sensitivity of the transgenic plants. In addition, our results shows that in the transgenic plants, the lower phytate levels has led to an increment of divalent cations, of which a 1.6 fold increase in the iron concentration in milled rice seeds was noteworthy. This increase could be attributed to reduced chelation of divalent metal (iron) cations, which may correlate to higher iron bioavailability in the endosperm of rice grains.ConclusionThe present study evidently suggests that seed-specific silencing of MIPS in transgenic rice plants can yield substantial reduction in levels of phytic acid along with an increase in inorganic phosphate content. However, it was also demonstrated that the low phytate seeds had an undesirable diminution in levels of myo-inositol and ascorbate, which probably led to sensitiveness of seeds to abscisic acid during germination. Therefore, it is suggested that though MIPS is the prime target for generation of low phytate transgenic plants, down-regulation of MIPS can have detrimental effect on myo-inositol synthesis and related pathways which are involved in key plant metabolism.Electronic supplementary materialThe online version of this article (doi:10.1186/1939-8433-6-12) contains supplementary material, which is available to authorized users.

The Mutualistic Fungus Piriformospora indica Protects Barley Roots from a Loss of Antioxidant Capacity Caused by the Necrotrophic Pathogen Fusarium culmorum

Fusarium culmorum causes root rot in barley (Hordeum vulgare), resulting in severely reduced plant growth and yield. Pretreatment of roots with chlamydospores of the mutualistic root-colonizing basidiomycete Piriformospora indica (subdivision Agaricomycotina) prevented necrotization of root tissues and plant growth retardation commonly associated with Fusarium root rot. Quantification of Fusarium infections with a real-time polymerase chain reaction assay revealed a correlation between root rot symptoms and the relative amount of fungal DNA. Fusarium-infected roots showed reduced levels of ascorbate and glutathione (GSH), along with reduced activities of antioxidant enzymes such as superoxide dismutase, ascorbate peroxidase, GSH reductase, dehydroascorbate reductase, and monodehydroascorbate reductase. Consistent with this, Fusarium-infected roots showed elevated levels of lipid hydroperoxides and decreased ratios of reduced to oxidized forms of ascorbate and GSH. In clear contrast, roots treated with P. indica prior to inoculation with F. culmorum showed levels of ascorbate and GSH that were similar to controls. Likewise, lipid peroxidation and the overall reduction in antioxidant enzyme activities were largely attenuated by P. indica in roots challenged by F. culmorum. These results suggest that P. indica protects roots from necrotrophic pathogens, at least partly, through activating the plant's antioxidant capacity.

Human Genetic Variation Influences Vitamin C Homeostasis by Altering Vitamin C Transport and Antioxidant Enzyme Function

New evidence for the regulation of vitamin C homeostasis has emerged from several studies of human genetic variation. Polymorphisms in the genes encoding sodium-dependent vitamin C transport proteins are strongly associated with plasma ascorbate levels and likely impact tissue cellular vitamin C status. Furthermore, genetic variants of proteins that suppress oxidative stress or detoxify oxidatively damaged biomolecules, i.e., haptoglobin, glutathione-S-transferases, and possibly manganese superoxide dismutase, affect ascorbate levels in the human body. There also is limited evidence for a role of glucose transport proteins. In this review, we examine the extent of the variation in these genes, their impact on vitamin C status, and their potential role in altering chronic disease risk. We conclude that future epidemiological studies should take into account genetic variation in order to successfully determine the role of vitamin C nutriture or supplementation in human vitamin C status and chronic disease risk.

Drought and air warming affect the species-specific levels of stress-related foliar metabolites of three oak species on acidic and calcareous soil

Climate change as projected for Central Europe will lead to prolonged periods of summer drought and enhanced air temperature. Thus, forest management practices are required to take into account how species performance is adapted to cope with these climate changes. Oak trees may play a major role in future forests because of their relative drought-tolerance compared with other species like beech. Therefore, this study investigated the stress responses (i.e., anti-oxidants, free amino acids) in the leaves of three widely distributed oak species in Central Europe (i.e., Quercus robur L., Q. petraea [Matt.] Libel., Q. pubescens Willd.) to drought, air warming and the combination of drought plus air warming under controlled conditions after periods of spring drought, a short rewetting and summer drought. We quantified foliar levels of thiols, ascorbate, and free amino compounds in Q robur, Q. petraea and Q. pubescens. Our study showed that oak saplings had increased levels of γ-glutamylcysteine and total glutathione and proline with drought and air warming. Foliar ascorbate, glutathione disulfide and dehydroascorbic acid levels were not affected. The comparison of stress responses to drought and/or air warming between the three species showed higher foliar thiol levels in Q. robur and Q. pubescens compared with Q. petraea. For total and reduced ascorbic acid and γ-aminobutyric acid, the highest levels were found in Q. robur. In conclusion, our study showed that foliar anti-oxidant and free amino acid levels were significantly affected by drought plus air warming; however, this effect was species-dependent with the drought-tolerant species of Q. pubescens having the highest reactive oxygen species scavenging capacity among three tested oak species. Furthermore, stress responses as shown by increased levels of foliar anti-oxidants and free amino acids differ between calcareous and acidic soil indicating that the capacities of anti-oxidative defense and osmotic stress adjustment developed better on calcareous compared with acidic soil; however, this effect was metabolite- as well as species-specific.

Neurochemical Changes in the Rat Occipital Cortex and Hippocampus after Repetitive and Profound Hypoglycemia During the Neonatal Period: An Ex Vivo 1H Magnetic Resonance Spectroscopy Study

The brain of a human neonate is more vulnerable to hypoglycemia than that of pediatric and adult patients. Repetitive and profound hypoglycemia during the neonatal period (RPHN) causes brain damage and leads to severe neurologic sequelae. Ex vivo high-resolution (1)H nuclear magnetic resonance (NMR) spectroscopy was carried out in the present study to detect metabolite alterations in newborn and adolescent rats and investigate the effects of RPHN on their occipital cortex and hippocampus. Results showed that RPHN induces significant changes in a number of cerebral metabolites, and such changes are region-specific. Among the 16 metabolites detected by ex vivo (1)H NMR, RPHN significantly increased the levels of creatine, glutamate, glutamine, γ-aminobutyric acid, and aspartate, as well as other metabolites, including succine, taurine, and myo-inositol, in the occipital cortex of neonatal rats compared with the control. By contrast, changes in these neurochemicals were not significant in the hippocampus of neonatal rats. When the rats had developed into adolescence, the changes above were maintained and the levels of other metabolites, including lactate, N-acetyl aspartate, alanine, choline, glycine, acetate, and ascorbate, increased in the occipital cortex. By contrast, most of these metabolites were reduced in the hippocampus. These metabolic changes suggest that complementary mechanisms exist between these two brain areas. RPHN appears to affect occipital cortex and hippocampal activities, neurotransmitter transition, energy metabolism, and other metabolic equilibria in newborn rats; these effects are further aggravated when the newborn rats develop into adolescence. Changes in the metabolism of neurotransmitter system may be an adaptive measure of the central nervous system in response to RPHN.

P65: Nitrite after cardiac arrest has antioxidant effects reducing reperfusion cerebral hyperemia and subsequent caspase activation

We previously demonstrated nitrite mediated neuroprotection after asphyxial cardiac arrest (ACA). Reperfusion injury after successful return of spontaneous circulation (ROSC) has been linked to reactive oxygen species generation resulting in loss of cerebral autoregulation with early hyperemia, activation of caspases and delayed neuronal death. Having previously demonstrated nitrite’s cardiac antioxidant effects after cardiac arrest, we hypothesized that a similar mechanism may prevent loss of autoregulation and subsequent caspase activation resulting in neuroprotection. Adult male Sprague–Dawley rats were anesthetized, intubated and catheterized (arterial and venous). An ultrasonic flow probe was placed around the ascending aorta to continuously measure cardiac output (CO) and cerebral cortex blood flow (CBF) was measured using laser speckle analysis through a 5 mm craniotomy. ACA was induced with paralysis and cessation of mechanical ventilation for 8 min prior to manual cardiopulmonary resuscitation with epinephrine and sodium bicarbonate. Animals were randomized 5 min post-ROSC to a 10 min 500 μl infusion of 8 μM nitrite or plasmalyte placebo. Temperature was maintained at 37 °C while heart rate (HR), blood pressure (BP), CO and CBF were measured for one hour and results are reported normalized to baseline values. At 1 h animals were perfused with saline, decapitated and brains homogenized for ascorbate and caspase measurements. In separate experiments without CO/CBF monitoring, animals were survived 24 h before brain homogenization for caspase analysis. Caspase 3/7 activity was measured using a commercial luminescent assay and ascorbate measured using a fluorescent assay with acridine TEMPO. All measurements were normalized to protein content of samples. Comparisons are by repeated measures ANOVA, 1 way ANOVA and Mann–Whitney U tests and mean (physiology, caspase) or median (ascorbate) values are reported below. Animals in both treatment arms had baseline HR, MAP, CO and CBF similar to each other and non-arrested shams and consistent with reported normal values ( Fig. 1 ). Sham physiologic values did not change significantly over 1 h of observation under isoflurane anesthesia. ACA resulted in complete loss of CO and CBF. Ten min after ROSC (5 min after treatment was initiated), CO was diminished (80%) in both treatment groups yet BP was significantly (138%) increased. In placebo animals, this resulted in a significant increase in CBF (187%) compared to nitrite treated animals (136%; p < 0.01). This difference persisted until 20 min after ROSC at which point groups converged. Nitrite treated animals had significantly higher levels of ascorbate (33.09 nmol/mg protein) compared to placebo treated animals (26.17; p = 0.027) 1 h after ROSC. At 24 h, ACA resulted in significant( p < 0.01) increases in caspase-3/7 activity compared to shams (1381 RLU/mg protein) with higher levels in placebo (5383) compared to nitrite (4340) treated animals. Nitrite acts as antioxidant when administered after ROSC blunting subsequent loss of autoregulation and hyperemia, preserving antioxidant reserve and reducing subsequent caspase-3/7 activation. This provides a mechanistic insight into nitrite mediated neuroprotection. Ongoing studies are examining potential subcellular targets for nitrite therapy which may explain these effects. Supported by NINDS K08.

Oxidative stress and chronic obstructive pulmonary disease: The impact of oral antioxidants on skeletal muscle fatigue

Oxidative stress may contribute to exercise intolerance in patients with COPD. This study sought to determine the effect of an acute oral antioxidant cocktail (AOC: vitamins C, E, and alpha‐lipoic acid) on skeletal muscle function in patients with COPD. Ten patients with COPD (FEV1/FVC &lt; 0.7, FEV1 ≤ 80% predicted) performed knee extensor exercise to exhaustion as well as isotime trials following either the AOC or placebo. Changes in quadriceps maximal voluntary contractions (MVCs) and potentiated twitch forces (Qtw,pot) quantified quadriceps fatigue. Despite increasing plasma ascorbate levels (10.1±2.2 to 24.1±3.8 ug/ml, p&lt;0.05) and attenuating the electron paramagnetic resonance (EPR) spectroscopy free radical signal (AUC: 11.6±3.7 to 4.8±2.2 AU, p&lt;0.05), AOC consumption did not alter endurance time or the development of quadriceps fatigue. Revaluation of the EPR spectroscopy data revealed that the AOC was most efficacious in patients with high (n=5, AUC: 19.7±5.8 to 5.8±4.5 AU) compared to low (n=4, AUC: 1.6±0.5 to 3.4±1.1 AU) basal free radical levels. This baseline index of free radicals was inversely correlated with FEV1 (r=−0.54, p&lt;0.05) and MVC force (r=−0.56, p&lt;0.05). Thus, while a potentially beneficial role for the AOC in COPD can be inferred, an acute reduction in free radicals does not appear to influence exercise performance in COPD. Supported by the National Institutes of Health grant PO1 HL 09830.

Oxidative Responses Induced by Pharmacologic Vitreolysis and/or Long-term Hyperoxia Treatment in Rat Lenses

ABSTRACTPurpose: The aim of the study was to investigate the protective effects of intact vitreous gel on the lens after pharmacologic vitreolysis and hyperoxia exposure in rats in vivo.Methods: Eyes of Sprague-Dawley rats were induced to posterior vitreous detachment (PVD) by pharmacologic vitreolysis, and the rats with and without PVD were treated with hyperoxia 3 h per day for 5 months. Lens transparency was monitored by a slit-lamp biomicroscope. A series of biochemical measurements were made in extracts of the lens cortex and nucleus. Ascorbate levels were measured in the aqueous and vitreous humors.Results: No significant differences in lens transparency or morphology were observed in all groups, and no significant biochemical changes were observed in the cortex or nucleus of lenses of the PVD group. In the lens nucleus, the values of water-soluble protein concentration in PVD + hyperoxia group were lower than that of the PVD group. The levels of water-soluble proteins, glutathione (GSH) and ascorbate decreased in the hyperoxia group with an intact vitreous body. Vitreolysis enhanced the effect of hyperoxia, decreasing soluble protein, GSH and ascorbate below the levels seen in eyes with vitreolysis alone. The levels of antioxidants and soluble proteins were lower in the lens nucleus, and the effects of vitreolysis plus hyperoxia were more significant in the nucleus. Hyperoxia and hyperoxia plus vitreolysis reduced catalase activity and increased oxidized GSH to a greater extent in the lens cortex, although these treatments increased protein-GSH mixed disulfides in both regions. Long-term hyperoxia also lowered ascorbate levels in the vitreous and aqueous humors, an effect that was enhanced by vitreolysis.Conclusions: Exposure to excess molecular oxygen produces significant oxidative damage to the lens, especially the lens nucleus. These effects were enhanced by pharmacologic vitreolysis, indicating that intact vitreous gel protects the lens from oxidative damage.

1H NMR metabolomics investigation of an Alzheimer’s disease (AD) mouse model pinpoints important biochemical disturbances in brain and plasma

In this study data generated by 1H NMR were combined with chemometrics to analyse brain and plasma samples from APP/PS1 and wild type mice with the aim of developing a statistical model capable of predicting the features of Alzheimer’s disease (AD) displayed by this animal model. APP/PS1 is a well characterised double transgenic mouse model of AD and the results here demonstrate the potential of NMR technology as a platform for the detecting this disease. Using partial least squares discriminant analysis a model was built using both brain extracts (R2 = 0.99; Q2 = 0.66) and a high throughput method of plasma analysis (R2 = 0.98; Q2 = 0.75) capable of predicting AD in APP/PS1 mice. Analysis of brain extracts led to the elucidation of 20 metabolites and 16 of these were quantifiable. Relative brain levels of ascorbate, creatine, γ-aminobutyric acid and N-acetyl aspartic acid were significantly altered in APP/PS1 mice (p < 0.05). Analysis of plasma identified 14 metabolites and the levels of acetate, citrate, glutamate, glutamine, methionine, and an unknown signal were significantly altered in APP/PS1 mice (p < 0.05). Combining 1H NMR spectral data with chemometrics has been previously used to study biochemical disturbances in various disease states. This study further indicates the translational potential of this technology for identifying AD in people attending the memory clinic.

Effect of Hyperoxia on Resuscitation of Experimental Combined Traumatic Brain Injury and Hemorrhagic Shock in Mice

Hypotension and hypoxemia worsen traumatic brain injury outcomes. Hyperoxic resuscitation is controversial. The authors proposed that hyperoxia would improve hemodynamics and neuronal survival by augmenting oxygen delivery despite increased oxidative stress and neuroinflammation in experimental combined controlled cortical impact plus hemorrhagic shock in mice. Adult C57BL6 mice received controlled cortical impact followed by 35 min of hemorrhagic shock (mean arterial pressure, 25-27 mmHg). The resuscitation phase consisted of lactated Ringer's boluses titrated to mean arterial pressure greater than 70 mmHg. Definitive care included returning shed blood. Either oxygen or room air was administered during the resuscitation phases. Brain tissue levels of oxidative stress and inflammatory markers were measured at 24 h and hippocampal neuronal survival was quantified at 7 days. Hyperoxia markedly increased brain tissue oxygen tension approximately four- to fivefold (n = 8) and reduced resuscitation fluid requirements approximately 15% (n = 53; both P < 0.05). Systemic and cerebral physiologic variables were not significantly affected by hyperoxia. Hippocampal neuron survival was approximately 40% greater with oxygen versus room air (n = 18, P = 0.03). However, ascorbate depletion doubled with oxygen versus room air (n = 11, P < 0.05). Brain tissue cytokines and chemokines were increased approximately 2- to 20-fold (n = 10) after combined controlled cortical impact injury plus hemorrhagic shock, whereas hyperoxia shifted cytokines toward a proinflammatory profile. Hyperoxic resuscitation of cortical impact plus hemorrhagic shock reduced fluid requirements and increased brain tissue oxygen tension and hippocampal neuronal survival but exacerbated ascorbate depletion and neuroinflammation. The benefits of enhanced oxygen delivery during resuscitation of traumatic brain injury may outweigh detrimental increases in oxidative stress and neuroinflammation.

Ontogenetic changes in vitamin C in selected rice varieties

Vitamin C (L-ascorbic acid) is a key antioxidant for both plants and animals. In plants, ascorbate is involved in several key physiological processes including photosynthesis, cell expansion and division, growth, flowering, and senescence. In addition, ascorbate is an enzyme cofactor and a regulator of gene expression. During exposure to abiotic stresses, ascorbate counteracts excessive reactive oxygen species within the cell and protects key molecules, including lipids, proteins, and nucleic acids, from irreversible damage. In this study we focus on understanding how ascorbate levels are controlled in rice (Oryza sativa) during plant development and in response to light intensity and photoperiod. Our results indicate that in rice ascorbate metabolism follows a different pattern compared to other species. In the rice accessions we analyzed, total foliar ascorbate content increases during development and peaks at the vegetative 2-4 and the reproductive 4 stages, whereas other research has shown that in Arabidopsis thaliana and other dicots, ascorbate content declines with plant age. The pattern in rice does not seem to change when plants were grown under increasing light intensity: 150, 400 or 1200-1500 μmol m(-2) s(-1). We observed little diurnal variation in AsA content in rice and did not see a steady decline during the dark period as has been reported in other species such as Arabidopsis and tomato. The total foliar ascorbate content of twenty-three rice accessions from four major rice subgroups was compared. These genotypes differed as much as eight-fold in ascorbate content at the V2 stage indicating the potential to enhance vitamin C levels in genotypes of global interest via breeding approaches.

The Inter-Relationship of Ascorbate Transport, Metabolism and Mitochondrial, Plastidic Respiration

Ascorbate, this multifaceted small molecular weight carbohydrate derivative, plays important roles in a range of cellular processes in plant cells, from the regulation of cell cycle, through cell expansion and senescence. Beyond these physiological functions, ascorbate has a critical role in responses to abiotic stresses, such as high light, high salinity, or drought. The biosynthesis, recycling, and intracellular transport are important elements of the balancing of ascorbate level to the always-changing conditions and demands. A bidirectional tight relationship was described between ascorbate biosynthesis and the mitochondrial electron transfer chain (mETC), since L-galactono-1,4-lactone dehydrogenase (GLDH), the enzyme catalyzing the ultimate step of ascorbate biosynthesis, uses oxidized cytochrome c as the only electron acceptor and has a role in the assembly of Complex I. A similar bidirectional relationship was revealed between the photosynthetic apparatus and ascorbate biosynthesis since the electron flux through the photosynthetic ETC affects the biosynthesis of ascorbate and the level of ascorbate could affect photosynthesis. The details of this regulatory network of photosynthetic electron transfer, respiratory electron transfer, and ascorbate biosynthesis are still not clear, as are the potential regulatory role and the regulation of intracellular ascorbate transport and fluxes. The elucidation of the role of ascorbate as an important element of the network of photosynthetic, respiratory ETC and tricarboxylic acid cycle will contribute to understanding plant cell responses to different stress conditions.

Cross Protection by Cold-shock to Salinity and Drought Stress-induced Oxidative Stress in Mustard (&lt;i&gt;Brassica campestris&lt;/i&gt; L.) Seedlings

In the present study, cold-shock (6 ℃ , 5.5 h) induced salinity and drought tolerance and involvement of antioxidative and glyoxalase systems were investigated in mustard ( Brassica campestris L.) seedlings. Seven-day-old seedlings were subjected to salt (150 mmol/L NaCl, 48 h) and drought stress (induced by 20% PEG, 48 h) with or without cold pre-treatment. The results showed that both salt and drought stresses abruptly increased the hydrogen peroxide (H 2 O 2 ) and lipid peroxidation (malondialdehyde, MDA) levels. Ascorbate (AsA), reduced glutathione (GSH) and oxidized glutathione (GSSG) contents, GSH/GSSG ratio and the activities of ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), glutathione reductase (GR), glutathione S -transferase (GST), glutathione peroxidase (GPX), catalase (CAT), glyoxalase I (Gly I), and glyoxalase II (Gly II) showed both homogeneity and discrepancies in the responses of mustard seedlings to salinity and drought stresses. Drought stress treatment resulted in a significant increases in AsA content. The GSH and GSSG content increased in response to both salt and drought stresses, however, the GSH/GSSG ratio decreased significantly in response to drought stress. Salt stress treatment resulted in a significant increase of APX, MDHAR, GR, GST and Gly I activities, whereas, CAT and Gly II activities decreased. In contrast, drought stress treatment resulted in a significant increase in MDHAR, DHAR, GPX and Gly I activities; whereas, APX, CAT and Gly II activities decreased. Importantly, cold pre-treated salt and drought-stressed seedlings maintained higher level of AsA, GSH contents and GSH/GSSG ratio, higher activities of APX, DHAR, GR, GST, GPX, CAT, Gly I and Gly II, and lower the levels of GSSG, H 2 O 2 and MDA as compared to the control as well as in most cases seedlings subjected to salt and drought stress without cold pre-treatment. Our findings showed that a retention of the imprint of previous stress exposure (short-term cold-shock), induces salt and drought-induced oxidative stress tolerance by modulating antioxidative and glyoxalase systems.

Clinical experience with intravenous administration of ascorbic acid: achievable levels in blood for different states of inflammation and disease in cancer patients.

BackgroundAscorbic acid (vitamin C, ascorbate) is a key water soluble antioxidant that, when administered in doses well above its recommended dietary allowance, may have preventative and therapeutic value against a number of pathologies. The intravenous administration of high dose ascorbate (IVC) has increased in popularity among complementary and alternative medicine practitioners: thousands of patients received IVC, at an average dose of 0.5 g/kg, without significant side effects. While IVC may have a variety of possible applications, it has generated the most interest for its potential use in treating cancer.MethodsMedical records of patients with cancer treated with IVC at the Riordan Clinic were retrospectively reviewed. Cancer patients, for whom plasma ascorbate concentration data before and after treatment were available, along with C-reactive protein (CRP) measurements, were chosen for analysis.ResultsThe results of the analysis can be summarized as follows. IVC produces peak plasma ascorbate concentrations on the order of ten millimolars with lower peak plasma concentrations obtained in cancer patients as compared to healthy subjects. Cancer patients who are deficient in vitamin C prior to therapy tend to achieve lower plasma levels post infusion. High inflammation or tumor burdens, as measured by CRP or tumor antigen levels, tend to lower peak plasma ascorbate levels after IVC. When compared to patients with localized tumors, patients with metastatic tumors tend to achieve lower post infusion plasma ascorbate concentrations.ConclusionsThe data indicate that, while potentially therapeutic plasma ascorbate concentrations can be achieved with IVC, levels attained will vary based on tumor burden and degree of inflammation (among other factors). Evidence suggests that IVC may be able to modulate inflammation, which in turn might improve outcomes for cancer patients. IVC may serve as a safe, adjunctive therapy in clinical cancer care.

Antisense-mediated depletion of tomato GDP-l-galactose phosphorylase increases susceptibility to chilling stress

The GDP-l-galactose phosphorylase (GGP), which converts GDP-l-galactose to l-Gal-1-phosphate, is generally considered to be a key enzyme of the major ascorbate biosynthesis pathways in higher plants, but experimental evidence for its role in tomato is lacking. In the present study, the GGP gene was isolated from tomato (Solanum lycopersicum) and transient expression of SlGGP-GFP (green fluorescent protein) fusion protein in onion cells revealed the cytoplasmic and nucleus localization of the protein. Antisense transgenic tomato lines with only 50–75% ascorbate level of the wild type (WT) were obtained. Chilling treatment induced lower increase in AsA levels and redox ratio of ascorbate in antisense transgenic plants compared with WT plants. Under chilling stress, transgenic plants accumulated more malendialdehyde (MDA) and more O2−, leaked more electrolytes and showed lower maximal photochemical efficiency of PSII (Fv/Fm), net photosynthetic rate (Pn), and oxidizable P700 compared with WT plants. Furthermore, the antisense transgenic plants exhibited significantly higher H2O2 level and lower ascorbate peroxidase (APX) activity. Our results suggested that GGP plays an important role in protecting plants against chilling stress by maintaining ascorbate pool and ascorbate redox state.