Ascorbic acid in rat lung

Fifty percent of the ascorbic acid content of sliced rat lung was removed in 2 min by washing with Krebs-phosphate solution. Although no ascorbic acid was washed from an intact lung by perfusion of the pulmonary circulation, 30% of it was removed by washing out the air spaces. It is concluded that 50% of the ascorbic acid present in rat lung is present in the fluid lining of the air spaces.     

Extracellular ascorbic acid in lung

Fifty percent of the ascorbic acid content of sliced rat lung was released from tissue to the media within a few minutes by either washing or incubating the slices with Kreb-phosphate solution. Measurement of the lactate dehydrogenase and potassium content of the medium after incubating lung slices for 5 min showed that about 20% of the cells were damaged by slicing.Sephadex chromatography of tissue extracts prepared from washed lung slices showed that none of the ascorbic acid in these slices was bound to protein. Also, metabolic poisons were shown to deplete the ascorbic acid content of washed lung slices.Approx. 57% of the lung ascorbic acid of guinea pigs that had been supplemented with ascorbic acid and 78% of the lung ascorbic acid of ascorbic acid-deficient guinea pigs were found in the medium when lung slices from these animals were incubated with Krebs-phosphate solution.These results were taken to indicate the presence of an extracellular pool of ascorbic acid in lung which is maintained even during scurvy.     

Determination of brain gangliosides by determination of ganglioside stearic acid

A new method is described for the determination of brain gangliosides by measuring stearic acid, the chief acid of gangliosides, in an appropriately purified brain extract. The method includes extraction of tissue with chloroform-methanol, extraction of gangliosides from the extract with 0.1 M KCl, evaporation of the aqueous phase, methanolysis, and gas-liquid chromatography of the resultant methyl esters with a double internal standard. The method depends on the simple composition of ganglioside fatty acids (80% stearic acid) and allows determination of less than 0.05 micromole of gangliosides. Interfering lipids are removed from the ganglioside extract by washing with chloroform-methanol-water. The effects of contamination with nonlipid N-acetylneuraminic acid are avoided.     

DNA degradation and repair in human laryngeal carcinoma HEp-2 cells after combined exposure to vitamin B12b and ascorbic acid

The formation and accumulation of DNA fragments containing no more than 23,000 pairs of bases were observed under exposure of human larynx epidermoid carcinoma cells (Hep-2) to "chemical nuclease", oxycobalamin (vitamin B12b) and ascorbic acid (vitamin C). The obtained DNA damages were repaired more slowly than those induced by gamma-irradiation in the dose adequate to the level of DNA damages. DNA reparation was not revealed after washing the cells from vitamin B12b and ascorbic acid, and in the course of cell incubation with ascorbic acid. Vitamin B12b and ascorbic acid separately did not induce degradation of DNA. DNA damages induced by "chemical nuclease" action precede the cell death observed later.     

Ontogenetic characteristics of ascorbic acid binding by nerve tissue proteins

The highest ascorbic acid content has been found in a low-molecular fraction (m. w. less than 6000) of water-soluble proteins from brain tissue homogenates of rats, the lowest one--in a high-molecular fraction (m. w. about 65,000). The content of protein-bound ascorbic acid decreases during postnatal life. This process is accompanied by changes in the ratio of the reduced and oxidated forms of the ascorbic acid in favour of the latter.     

Effect of oxygen on ascorbic acid uptake and concentration in embryonic chick brain

The effects of oxygen on ascorbic acid concentration and transport were studied in chick embryo (Gallus gallus domesticus). During normoxic incubations, plasma ascorbic acid concentration peaked on fetal day 12 and then fell, before increasing again on day 20 when pulmonary respiration began. In contrast, cerebral ascorbic acid concentration rose after day 6, was maintained at a relatively high level during days 8–18, and then fell significantly by day 20. Exposure of day 16 embryos for 48 h to 42% ambient O₂ concentration decreased ascorbic acid concentration by four-fifths in plasma and by one-half in brain, compared to values in normoxic (21% O₂) or hypoxic (15% O₂) controls. Hyperoxic preincubation of embryos also inhibited ascorbic acid transport, as evidenced by decreased initial rates of saturable and Na⁺-dependent [¹⁴C]ascorbic acid uptake into isolated brain cells. It may be concluded that changes in ascorbic acid concentration occur in response to oxidative stress, consistent with a role for the vitamin in the detoxification of oxygen radicals in fetal tissues. However, changing O₂ levels have less effect on ascorbic acid concentration in brain than in plasma, indicating regulation of the vitamin by brain cells. Furthermore, the effect of hyperoxia on cerebral vitamin C may result, in part, from inhibition of cellular ascorbic acid transport.     

Elevated oxidative stress and sensorimotor deficits but normal cognition in mice that cannot synthesize ascorbic acid

Oxidative stress is implicated in the cognitive deterioration associated with normal aging as well as neurodegenerative disorders such as Alzheimer's and Parkinson's diseases. We investigated the effect of ascorbic acid (vitamin C) on oxidative stress, cognition, and motor abilities in mice null for gulono-γ-lactone oxidase (Gulo). Gulo−/− mice are unable to synthesize ascorbic acid and depend on dietary ascorbic acid for survival. Gulo−/− mice were given supplements that provided them either with ascorbic acid levels equal to- or slightly higher than wild-type mice (Gulo-sufficient), or lower than physiological levels (Gulo-low) that were just enough to prevent scurvy. Ascorbic acid is a major anti-oxidant in mice and any reduction in ascorbic acid level is therefore likely to result in increased oxidative stress. Ascorbic acid levels in the brain and liver were higher in Gulo-sufficient mice than in Gulo-low mice. F₄-neuroprostanes were elevated in cortex and cerebellum in Gulo-low mice and in the cortex of Gulo-sufficient mice. All Gulo−/− mice were cognitively normal but had a strength and agility deficit that was worse in Gulo-low mice. This suggests that low levels of ascorbic acid and elevated oxidative stress as measured by F₄-neuroprostanes alone are insufficient to impair memory in the knockouts but may be responsible for the exacerbated motor deficits in Gulo-low mice, and ascorbic acid may have a vital role in maintaining motor abilities.     

Vitamin C is a source of oxoaldehyde and glycative stress in age‐related cataract and neurodegenerative diseases

Oxoaldehyde stress has recently emerged as a major source of tissue damage in aging and age‐related diseases. The prevailing mechanism involves methylglyoxal production during glycolysis and modification of arginine residues through the formation of methylglyoxal hydroimidazolones (MG‐H1). We now tested the hypothesis that oxidation of vitamin C (ascorbic acid or ASA) contributes to this damage when the homeostatic redox balance is disrupted especially in ASA‐rich tissues such as the eye lens and brain. MG‐H1 measured by liquid chromatography mass spectrometry is several fold increased in the lens and brain from transgenic mice expressing human vitamin C transporter 2 (hSVCT2). Similarly, MG‐H1 levels are increased two‐ to fourfold in hippocampus extracts from individuals with Alzheimer's disease (AD), and significantly higher levels are present in sarkosyl‐insoluble tissue fractions from AD brain proteins than in the soluble fractions. Moreover, immunostaining with antibodies against methylglyoxal hydroimidazolones reveals similar increase in substantia nigra neurons from individuals with Parkinson's disease. Results from an in vitro incubation experiment suggest that accumulated catalytic metal ions in the hippocampus during aging could readily accelerate ASA oxidation and such acceleration was significantly enhanced in AD. Modeling studies and intraventricular injection of ¹³C‐labeled ASA revealed that ASA backbone carbons 4–6 are incorporated into MG‐H1 both in vitro and in vivo, likely via a glyceraldehyde precursor. We propose that drugs that prevent oxoaldehyde stress or excessive ASA oxidation may protect against age‐related cataract and neurodegenerative diseases.     

ISOLATION AND DETERMINATION OF NON-DIFFUSIBLE SIALOFUCOHEXOSAMINOGLYCANS DERIVED FROM BRAIN GLYCOPROTEINS AND THEIR ANATOMICAL DISTRIBUTION IN BOVINE BRAIN

Abstract— Glycoproteins in brain tissue were assayed by determining the amount of N-acetylneuraminic acid (NANA), hexosamine, hexose, and fucose present in glycopeptides released by the proteolytic action of papain on the defatted protein residue that remains after treatment of the sample with chloroform-methanol (2:1 and 1:2, v/v). Diffusible and non-diffusible glycopeptides (sialofucohexosaminoglycans) were released by proteolysis. The procedure demonstrated that successive treatment of brain tissue with chloroform-methanol (2:1, v/v) and chloroform-methanol (1:2, v/v) removed all of the gangliosides present in the tissue. A 1 hr autolysis of rat brain tissue had no effect on the amount of glycopeptides recovered from the tissue. The carbohydrate composition of the non-diffusible sialofucohexosaminoglycans was also unaffected. Areas of the brain that are enriched in neuronal cell bodies contained a higher concentration of gangliosides and glycoproteins than areas that consist largely of myelinated fibre tracts. On the other hand, there was a greater concentration of glycoprotein relative to that of gangliosides in areas that consist predominately of myelinated fibre tracts and glia than in areas enriched in neuronal cell bodies. The concentration of non-diffusible sialofucohexosaminoglycans in whole bovine brain was less than that in whole rat brain. The non-diffusible sialofucohexosaminoglycans from whole bovine brain contained less fucose and NANA per mole of hexosamine and hexose than non-diffusible sialofucohexosaminoglycans isolated from whole rat brain. The non-diffusible sialofucohexosaminoglycans isolated from bovine cerebral white matter were lower in fucose and NANA content per mole of hexose and hexosamine than those isolated from other brain areas. It is suggested that the fucose and NANA content of the non-diffusible sialofucohexosaminoglycans associated with myelinated axons and (or) glia is less than that of the non-diffusible sialofucohexosaminoglycans associated with the nerve cell body.     

Ascorbic acid status of mullet, Mugil cephalus Linn., exposed to cadmium

The effects of acute and chronic cadmium exposures on ascorbic acid concentrations and cadmium accumulation in the tissues of juvenile mullet, Mugil cephalus , are described. Hepatic ascorbic acid reserves were maintained in untreated mullet fed a diet containing ascorbate, but decreased 60% in individuals exposed to 10 mg Cd 1^-1 for six weeks. Ascorbic acid depletion was accompanied by a massive accumulation of cadmium in the liver. Similar changes were found in gill tissue. In contrast, ascorbic acid concentrations were largely unaffected in kidney tissue which had a limited capacity to accumulate the metal. There were marked fluctuations of ascorbic acid levels in the brain. Only small amounts of cadmium were accumulated by this tissue. These results indicate that in mullet liver, gill and brain ascorbic acid stores may be depleted during chronic cadmium poisoning.     

Capacitive detection of glucose using molecularly imprinted polymers

A novel glucose biosensor based on capacitive detection has been developed using molecularly imprinted polymers. The sensitive layer was prepared by electropolymerization of o-phenylenediamine on a gold electrode in the presence of the template (glucose). Cyclic voltammetry and capacitive measurements monitored the process of electropolymerization. Surface uncovered areas were plugged with 1-dodecanethiol to make the layer dense, and the insulating properties of the layer were studied in the presence of redox couples. The template molecules and the nonbound thiol were removed from the modified electrode surface by washing with distilled water. A capacitance decrease could be obtained after injection of glucose. The electrode constructed similarly but with ascorbic acid or fructose only showed a small response compared with glucose. The stability and reproducibility of the biosensor were also investigated.     

Comparative tissue ascorbic acid studies in fishes

Comparative tissue ascorbic acid levels in four species of major carp viz., Labeo rohila, L. calbasu, Cirrhina tnrigala and Catla catla , were investigated. The ascorbic acid level was found to be the highest in the spleen in the four species studied (range 430–380 μg/g) followed by the anterior (adrenal) kidney, gonads, liver, renal kidney, brain and/or eye. Heart and blood had the lowest levels (range 26–18 μg/ml) amongst the tissues studied. Overall tissue ascorbic acid levels were the highest in L. rohita and the lowest in C. mrigala . Investigation on seasonal variations in blood and kidney ascorbic acid levels of Notopterus notopterus revealed peak levels in spring (February-April) and the lowest levels in the postspawning period (August-September).     

Replacement time for alveolar lipid removed by pulmonary lavage: effects of multiple lavage on lung lipids.

Daily washing in vivo of the lung with 0.15 M saline did not deplete the Beagle dog lung of surfactant lipids, but rather increased the quantity of surfactant lipid in the tissue. Replacement time for the lung lipids removed by the lavage was approximately 5 hours. This rate is one indication of the time required for movement of surfactant lipid from storage areas to the surface of the alveoli. The increase in tissue surfactant lipid following multiple lavage suggests that the rate of surfactant lipid synthesis is controlled in part by the level of surfactant lipid in the alveoli.     

Redox Changes in Perfusates Following Intracerebral Penetration of Microdialysis Probes

Microdialysis probe insertion into rat cerebral cortex significantly affects the levels of redox-active substances in brain extracellular fluid. Ascorbic acid levels are high immediately after probe insertion, decline rapidly, and then rise as the rat recovers from anesthesia 5–8 hours after surgery. Uric acid is at a low level for 5 hours and then rapidly increases in parallel with ascorbic acid. High ascorbic acid levels immediately after probe insertion are likely due to a shift from intracellular to extracellular fluids, whereas the delayed increase in uric acid may be due to increased enzymatic formation. After removal from the brain, hydrogen peroxide (H₂O₂) in microdialysis samples produces catalase-sensitive oxidative chemiluminescence. Microdialysis samples also produce high level catalase-resistant chemiluminescence associated with ascorbic acid levels after penetration injury. Although ascorbic acid is likely an antioxidant at concentrations estimated to be in brain extracellular fluid, it may have prooxidant effects when complexed with transition metals released into the neuronal microenvironment during traumatic brain injury.     

Ascorbic acid, cognitive function, and Alzheimer's disease: a current review and future direction

This narrative review appraises the human and animal studies implicating ascorbic acid (AA) in normal cognitive function and Alzheimer's disease. A research framework for how nutrition affects brain aging is proposed with emphasis on AA intake, status, metabolism, and transport into brain tissue. A final synopsis highlights areas for future research regarding AA nourishment and healthy brain aging.     

Ascorbic acid and hydroxyproline levels in the serum and granulation tissue of rats with aseptic and infected wounds

Hydroxyproline and ascorbic acid were measured in wound tissues and ascorbic acid was determined in the blood sera of 230 mature male Wistar rats with aseptic and infected surface wounds on days 1-10 (daily), 12, and 15. The principal morphologic characteristics of the granulation tissue were assessed by semiquantitative methods. An infected wound is characterized by increased hydroxyproline levels in the granulation tissue and elevated ascorbic acid concentration in the blood serum. The granulation tissue ascorbic acid level augments during the first nine days in case of an aseptic wound and during twelve days in case of an infected wound. Morphologic and biochemical correlations are indicative of a relationship between the ascorbic acid level on the one hand and granulation tissue vascularization and fibroblast proliferation on the other.     

Synergistic inhibitory effect of ascorbic acid and acetylsalicylic acid on prostaglandin E2 release in primary rat microglia

Ascorbic acid (vitamin C) has been suggested to protect cerebral tissue in a variety of pathophysiological situations such as head trauma, ischemia or Alzheimer's disease. Most of these protective actions have been attributed to the antioxidative capacity of ascorbic acid. Besides the presence of elevated levels of oxygen radicals, prostaglandins produced by neurones and microglial cells seem to play an important role in prolonged tissue damage. We investigated whether ascorbic acid alone inhibits prostaglandin E2 (PGE2) synthesis and may augment the inhibitory effect of acetylsalicylic acid on prostaglandin synthesis. Ascorbic acid dose-dependently inhibited PGE2 synthesis in lipopolysaccharide-treated primary rat microglial cells (IC50 = 3.70 micro m). In combination with acetylsalicylic acid (IC50 = 1.85 micro m), ascorbic acid augmented the inhibitory effect of acetylsalicylic acid on PGE2 synthesis (IC50 = 0.25 micro m in combination with 100 micro m ascorbic acid). Ascorbic acid alone or in combination with acetylsalicylic acid did not inhibit cyclooxygenase-2 (COX-2) protein synthesis but inhibited COX-2 enzyme activity. Our results show that ascorbic acid and acetylsalicylic acid act synergistically in inhibiting PGE2 synthesis, which may help to explain a possible protective effect of ascorbic acid in various brain diseases.     

Ascorbate- and dehydroascorbic acid-mediated reduction of free radicals in the human erythrocyte

Nitroxides were used as models of persistent free radicals to study the antioxidant function of ascorbic acid in the human erythrocyte. It was concluded that: 1) ascorbate and other reductant(s) derived from dehydroascorbic acid (DHA) in the presence of thiols are the only significant reducing agents for nitroxides, 2) glutathione and DHA reduce nitroxides by a process that cannot be inhibited by ascorbic acid oxidase, 3) erythrocytes can be depleted of ascorbic acid by exhaustive washing in the presence of membrane-permeable cationic nitroxides such as N,N-dimethylamino-Tempo, 4) ascorbate-depleted cells do not reduce nitroxides; however, nitroxide reduction is restored when the cells are incubated with DHA, 5) reduction of nitroxides in ascorbate-depleted, DHA-treated cells is significantly faster than in buffered solutions of DHA and glutathione, 6) several equivalents of nitroxide are reduced relative to the intracellular ascorbate pool, 7) sustained nitroxide reduction is observed even when most of the intracellular ascorbate is oxidized, 8) spin trapping of oxyradicals in tert-butyl hydroperoxide-treated cells is accelerated with ascorbate depletion and inhibited with ascorbate loading, 9) ascorbate can be quantified within intact cells by analyzing the initial reduction rates of membrane-permeable cationic nitroxides, and 10) DHA-stimulated reduction of cationic nitroxides is slower and less extensive in erythrocytes deficient in glucose-6-phosphate dehydrogenase than in normal erythrocytes.     

Formation of a persistent inhibitory state of brain adenylate cyclase by GTP analog: Regional differences

Inhibition of adenylate cyclase by pretreatment with guanyl-5′-ylimidodiphosphate (10 μM) at 0°C was examined in particulates obtained from 8 non-neural tissues and from various areas of the CNS of the guinea pig. The excess of the GTP analog used for the pretreatment was removed by washing prior to the enzyme assay. Under these conditions, adenylate cyclase from all the non-neural tissues was activated or not altered while that from the brain was persistently inhibited with kinetics properties similar to those reported previously (Yamamoto and Shimizu, 1983). In the CNS, in general, the inhibition was more marked in those areas containing high densities of neuronal plasma membrane (cerebral cortex, cerebellum, hippocampus) than in areas consisting predominantly of myelin and glial cells (pons, medulla, spinal cord). The formation of a persistently inhibitory state presented herein represents one of the properties specific to brain adenylate cyclase.     

Seed Removal Rates Under Isolated Trees and Continuous Vegetation in Semiarid Thornscrub

Clearing native vegetation to increase the amount of land available for agriculture in northeastern Mexico has left remnants ranging in size from fragments of continuous vegetation to isolated individual trees. These provide valuable opportunities for restoring larger areas of native vegetation. We explored whether fragmentation of Tamaulipan thornscrub affects the removal of seeds from 12 woody species that encompass a range of sizes and dispersal mechanisms. We tested whether (1) seed removal rates under isolated trees were higher than under continuous vegetation; (2) dispersal structures, such as fleshy pericarps, made some seeds more attractive to seed removers; and (3) microenvironmental variation affected seed removal rates. Seeds were placed under canopies of Texas ebony (Ebenopsis ebano) and Mesquite trees (Prosopis laevigata). Seed removal trials were conducted three times, each trial lasting 30 days. Most seeds were removed in all trials by the end of one month. Seed removal rate was slower under isolated trees. In general, fleshy fruits were removed faster than other fruits; whole fruits and fleshy tissue were removed faster than depulped seeds. In species with fleshy pericarps, acid washing of seeds, to simulate seeds processed in the digestive tract of dispersers, reduced the seed removal rates, suggesting that it would be a good pre‐treatment for restoration efforts.