M Levine's research while affiliated with National Institutes of Health and other places

Publications (54)

Article
Full-text available
Ascorbic acid is a widely used and controversial alternative cancer treatment. In millimolar concentrations, it is selectively cytotoxic to many cancer cell lines and has in vivo anticancer activity when administered alone or together with other agents. We carried out a dose-finding phase I and pharmacokinetic study of i.v. ascorbic acid in patient...
Poster
SVCT1 (Slc23a1) knockout mice: Slc23a1 as the vitamin C kidney reabsorptive transporter
Chapter
Vitamin C is an electron donor, and this property accounts for its known and postulated functions. As an antioxidant, or reducing agent, the vitamin sequentially donates two electrons from the C2-C3 double bond. The first intermediate, formed by the loss of one electron, is the unstable free radical semidehydroascorbic acid. This intermediate is re...
Article
Full-text available
Editor—Ascorbate or vitamin C (VC) is an essential reducing agent and antioxidant that participates in a variety of metabolic processes.1 Unlike rodents and other animals, humans depend on dietary intake of VC to meet their daily requirement.1 2 VC intracellular accumulation is mediated through two distinct pathways. In one pathway, vitamin C is tr...
Poster
Dehydroascorbic acid (DHA) is reduced in human neutrophils by the glutathione(GSH)-dependent protein glutaredoxin. The mechanism could occur via an ascorbyl free radical (AFR) intermediate. The present studies investigate the AFR appearance during enzymatic DHA reduction. Electron spin resonance spectroscopy (ESR) was used to determine the ascorbyl...
Presentation
Ascorbate accumulation in human neutrophils is controlled by two mechanisms. The first mechanism is via a sodium-dependent ascorbate transporter. Two isoforms of human ascorbate transporters were cloned and expressed in Xenopus laevis ooctyes. The second mechanism is ascorbate recycling, a three-step process consisting of oxidation of extracellular...
Chapter
The current recommended dietary allowance in the United States (U.S. RDA) for vitamin C (ascorbate) is 60 mg daily [1, 2]. Similar or lower values are used in other countries throughout the world except France, where the value is 80 mg daily. The U.S. RDA is based on prevention of clinical vitamin C deficiency, or scurvy, with a margin of safety. S...
Article
To conduct an inpatient study on the recommended dietary allowance (RDA) for vitamin C, we developed a unique vitamin C-deficient diet using a nutrient database and selective menus. Fourteen different menus were developed offering > 300 items with 0-2.4 mg vitamin C per serving. During the 4-6 mo volunteers were hospitalized, daily dietary vitamin...
Article
Full-text available
Determinants of the recommended dietary allowance (RDA) for vitamin C include the relationship between vitamin C dose and steady-state plasma concentration, bioavailability, urinary excretion, cell concentration, and potential adverse effects. Because current data are inadequate, an in-hospital depletion-repletion study was conducted. Seven healthy...
Article
Full-text available
Although the recommended dietary allowance provides an estimate for vitamin C ingestion in humans, optimal vitamin C requirements are unknown. We define optimal vitamin C requirements operationally based on the following: dose-function relations, availability in the food supply, steady state concentrations in plasma and tissues achieved at each dos...
Article
Ascorbic acid (vitamin C) transport was investigated in human B lymphocytes. The vitamin was transported by two components. The first was a high-affinity activity with an apparent Km of 7-10 microM and Vmax of 0.14 mM/h (3.11 x 10(-4) mumol x h-1 x mg protein-1). The activity was concentration and temperature dependent, saturable, and inhibited by...
Article
Full-text available
The effect of ascorbic acid on glucose-induced insulin release from single pancreatic islets was measured using a new, ultra-sensitive enzyme-linked immunosorbent insulin assay. Within 20 s ascorbic acid inhibited insulin secretion; inhibition was dose dependent and completely reversible. There was a 50% inhibition of the secretory response with 20...
Article
Optimal ascorbic acid (vitamin C) requirements in humans are unknown. In situ kinetics is a biochemical approach to determine requirements for vitamin C and other vitamins. In situ kinetics requires that cellular functions of ascorbic acid are characterized. Vitamin-C-dependent cellular reactions are directly related to vitamin C concentrations ins...
Article
Full-text available
As the initial step in the use of fibroblasts as a model system for 'in situ kinetics', ascorbic acid (vitamin C) accumulation in normal human fibroblasts was investigated for the first time. Ascorbic acid was transported into fibroblasts and accumulated against a concentration gradient up to 20-fold, as measured by h.p.l.c. with coulometric electr...
Article
Full-text available
Ascorbic acid (vitamin C) accumulation in activated human neutrophils is increased as much as 10-fold above the mM concentrations present in normal neutrophils. Internal concentrations as high as 14 mM are achieved when external vitamin is at physiologic concentration. The mechanism is by oxidation of external vitamin to dehydroascorbic acid, prefe...
Article
A new method for detection of hydroxyproline has been developed. Hydroxyproline was derivatized using 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole; separation and detection were accomplished using HPLC with coulometric electrochemical detection. Derivatized hydroxyproline was initially detected using a 16 channel coulometric electrochemical array system...
Article
Full-text available
Because of the structural similarity between glucose and ascorbic acid, we investigated the effect of glucose on uptake and accumulation of ascorbic acid in isolated normal human neutrophils. Ascorbic acid accumulation was determined using high-performance liquid chromatography with coulometric electrochemical detection, in conjunction with liquid...
Article
Full-text available
Parkinson's disease has been modeled in humans, lower primates, and to a lesser extent in some other vertebrates by administration of the potent neurotoxin MPTP (1-methyl-4-phenyl-1,2,3,6 tetrahydropyridine). The MPTP model has thus drawn considerable attention as a system to search for anti-Parkinson's disease drugs, although the cost and scarcity...
Article
Biochemical and clinical evidence indicate that in situ kinetics is the first quantitative and achievable approach for determining optimal requirements for ascorbic acid, and for other vitamins.
Article
Full-text available
Ascorbic acid requirements are based on preventing the deficiency disease scurvy and on urinary excretion of vitamin C. We proposed the first quantitative approach to determining optimal requirements for ascorbic acid and other vitamins, called in situ kinetics. In situ kinetics biochemically is based on the application of Michaelis-Menten reaction...
Article
Full-text available
The transport and accumulation of ascorbic acid in normal human skin fibroblasts in culture was investigated by using high-performance liquid chromatographic separation and coulometric electrochemical detection. Results measured as picomole ascorbic acid per microgram cell protein were expressed in molar amounts after determining the volume of skin...
Article
Full-text available
The uptake and distribution of ascorbic acid and the effect of extracellular glucose on ascorbic acid transport were investigated in human neutrophils. Freshly isolated neutrophils contained 1.0-1.4 mmol ascorbic acid/L, at least 94% of which was present unbound in the cytosol. Intracellular ascorbic acid was found only in the reduced form. The pre...
Article
We investigated whether circulating ascorbic acid in humans is protein bound or free and whether ascorbic acid exists in its reduced form alone as ascorbic acid or in its reduced and oxidized forms as ascorbic acid and dehydroascorbic acid, respectively. Ascorbic acid and dehydroascorbic acid were determined by using HPLC with coulometric electroch...
Article
Full-text available
We investigated whether semidehydroascorbic acid was an intermediate in norepinephrine synthesis in chromaffin granules and in electron transfer across the chromaffin granule membrane. Semidehydroascorbic acid was measured in intact granules by electron spin resonance. In the presence of intragranular but not extragranular ascorbic acid, semidehydr...
Article
Full-text available
We have investigated in intact chromaffin secretory vesicles the kinetics, specificity, and mechanism of intragranular ascorbic acid regeneration by extragranular ascorbic acid. The apparent Km of internal ascorbic acid regeneration for external ascorbic acid was 280 microM by Lineweaver-Burk analysis and 287 microM by Eadie-Hofstee analysis. Intra...
Article
Full-text available
Ascorbic acid (vitamin C) was found in isolated human mononuclear leukocytes and their purified components in millimolar concentration. Intracellular ascorbic acid was depleted greater than 96% during cell culture and was rapidly reaccumulated after addition of physiologic concentrations of ascorbic acid to the extracellular medium. Purified cells...
Article
Full-text available
The transport, accumulation, and distribution of ascorbic acid were investigated in isolated human neutrophils utilizing a new ascorbic acid assay, which combined the techniques of high performance liquid chromatography and coulometric electrochemical detection. Freshly isolated human neutrophils contained 1.0–1.4 mM ascorbic acid, which was locali...
Article
Ascorbic acid requirements for norepinephrine biosynthesis were investigated in intact bovine chromaffin granules using the physiologic substrate dopamine and a novel coulometric electrochemical detection high pressure liquid chromatography system for ascorbic acid. 10 mM external dopamine, 1 mM Mg-ATP, and 1 mM ascorbic acid produced maximal norep...
Article
Full-text available
Ascorbic acid requirements for norepinephrine biosynthesis were investigated in intact bovine chromaffin granules using the physiologic substrate dopamine and a novel coulometric electrochemical detection high pressure liquid chromatography system for ascorbic acid. 10 mM external dopamine, 1 mM Mg-ATP, and 1 mM ascorbic acid produced maximal norep...
Article
To the Editor: In the excellent article by Weiss on tissue destruction by neutrophils (Feb. 9 issue),¹ there is an implicit assumption that neutrophils lack endogenous reducing substances that could quench intracellular or extracellular oxidants. We would therefore like to emphasize that ascorbic acid is present in human neutrophils, in concentrati...
Article
Full-text available
Barium ions enter chromaffin cells via voltage-sensitive calcium channels, although the intracellular site of barium action is distinct from that of calcium. The entry of barium primarily through voltage-sensitive channels was indicated by experiments showing inhibition of 133Ba2+ uptake by blockers of voltage-dependent calcium channels. In additio...
Article
Full-text available
Ascorbic acid and Mg-ATP were found to regulate norepinephrine biosynthesis in intact secretory vesicles synergistically and specifically, using the model system of isolated bovine chromaffin granules. Dopamine uptake into chromaffin granules was shown to be unrelated to the presence of Mg-ATP and ascorbic acid at external dopamine concentrations o...
Article
Full-text available
The effect of ascorbic acid on the conversion of dopamine to norepinephrine was investigated in isolated chromaffin granules from bovine adrenal medulla. Ascorbic acid was shown to double the rate of [3H]norepinephrine formation from [3H]dopamine, despite no demonstrable accumulation of ascorbic acid into chromaffin granules. The enhancement of nor...
Article
Full-text available
The subcellular localization of catecholamines and ascorbic acid in cultured bovine adrenal chromaffin cells was studied by permeabilizing the cells with digitonin, a steroid glycoside. Catecholamine release from permeabilized chromaffin cells was dependent on the free calcium concentration and the temperature of the incubation mixture. By contrast...
Article
Full-text available
Ascorbic acid donates electrons to dopamine beta-monooxygenase during the hydroxylation of dopamine to norepinephrine in vitro. However, the possible role of ascorbic acid in norepinephrine biosynthesis in vivo has not been defined. We therefore investigated the effect of newly accumulated ascorbic acid on catecholamine biosynthesis in cultured bov...
Article
Full-text available
Ascorbic acid was found to be secreted from cultured bovine adrenal chromaffin cells coincidentally with catecholamines using a variety of secretagogues, including veratridine, nicotine, acetylcholine, and potassium chloride. Secretion of ascorbic acid was measured by preloading the cells for 3 h with (R)-[14C]ascorbic acid and then quantitating re...
Article
Full-text available
Catecholamines are transported into chromaffin granules by a Mg2+/ATP-driven process under conditions in which the substrate exists primarily as a positively charged or neutral species. In order to distinguish between these two states, we studied the transport properties of a permanently charged quaternary analogue of epinephrine, (R,S)-dimethylepi...

Citations

... It accumulates in skin cells, stabilizes collagen and enhances wound healing [17]. In skin cells as well as immune cells such as neutrophils and macrophages, vitamin C is taken up by sodium-dependent vitamin C transporter 1/2 (SVCT1/2), suggesting an important role of vitamin C in those cells [18,19]. It is known that vitamin C promotes leukocyte chemotaxis and reactive oxygen species (ROS) generation during phagocytosis and enhances apoptosis [17]. ...
... Vitamin C is a potent anti-oxidant which protects the body against oxidative stress (Padayatty et al., 2003). Stress, trauma or injury can use up large quantities of vitamin C (Long, 2003). ...
... In airway diseases such as CF and COPD, poorly controlled inflammation can result in a severe protease-antiprotease imbalance, largely due to the increased activities of NE. NE is a potent enzyme capable of degrading components of connective tissue, including elastin [49,50]. HS and heparin, and to a lesser extent HA and CS, may provide protection to the structural integrity of the lungs by strongly inhibiting leukocyte elastase activity and preventing breakdown of elastin in the lung [51]. ...
... 90 mg 2000 mg (Hemilä and Chalker, 2020;Institute of Medicine et al., 2002;Levine et al., 1998) Vitamin D Oily fish, liver, eggs, fortified foods (spreads and some breakfast cereals) ...
... tamin vizeletbe történő ürítésének mértékét [5]. Ezzel párhuzamosan a C-vitamin vérszintje 50–70%-kal csökkent homozigóta mutánsok esetén a vad típusú társakhoz képest. ...
... This data information could recommend the cherry tomato lines which could be consumed to fulfill and overcome sufficiency vitamine A and C. Necessity of vitamine C for daily is around 60 mg [30], but there is a variation of each individual. Some cases of phisical stress such as burns, infection, heavy metal poisoning, smoking, constantly drugs, etc can increase the body's necessity for vitamine C. Furthermore the smookers need vitamin C around 100 mg per daily [25]. In the Table 2 could be informed that the preferred cherry tomato lines by 50 respondents were IJR 154, IJR 151, IJR 1158, IJR 1150, local genotype, IJR 2170 respectivelly. ...
... The enzymatic and non-enzymatic functions of vitamin C were reviewed by Levine. Its role as an antioxidant has been extensively examined, in addition many vitamin C effects appear to be due to its role as a coenzyme in many biochemical reactions [9]. ...
... • The remaining three have the following functions: o dopamine beta hydroxylase participates in the biosynthesis of norepinephrine from dopamine (Levine, 1992;Kaufman, 1974). ...
... [37] Usually, in lower vertebrates, MPTP could not elicit PD-like movement disorder as the structure similar to SN in the teleost midbrain is absent. The nervous system of the goldfish is simple and has a highly lipophilic blood-brain barrier that helps in the uptake of majority neurotrophic and neuroprotective compounds; accessible for the study of PD. [38] Studies performed by Brodie et al. and Nagatsu et al. have reported the existence of serotonin, noradrenaline, and tyrosine hydroxylase in the forebrain and midbrain regions of goldfish. Further, Youdim and his coworkers have reported the occurrence of monoamino oxidases (MAO) A and B, DA, and noradrenaline (NA) in the fore, midbrain, and vagal lobes of goldfish. ...
... The pyruvate concentration in human plasma is around 100 μM, and this concentration increases after glucose uptake because pyruvate is a major product of glycolysis [35,58,59]. Furthermore, glucose was shown to inhibit intestinal vitamin C transport ex vivo [60], and blood glucose levels may interfere with the uptake of ascorbate by human neutrophils [61]. These observations suggest that considering adequate dietary patterns is critical when delivering AA supplements orally. ...