DNA damage from micronutrient deficiencies is likely to be a major cause of cancer

Mutation Research 475 (2001) 7–20
Bruce N. Ames
University of California, Berkeley, CA 94720-3202, USA
Received 11 May 2000;
received in revised form 10 August 2000;
accepted 8 November 2000


A deficiency of any of the micronutrients: folic acid, Vitamin B12, Vitamin B6, niacin, Vitamin C, Vitamin E, iron, or zinc, mimics radiation in damaging DNA by causing single- and double-strand breaks, oxidative lesions, or both. For example, the percentage of the US population that has a low intake (<50% of the RDA) for each of these eight micronutrients ranges from 2 to >20%.

A level of folate deficiency causing chromosome breaks was present in approximately 10% of the US population, and in a much higher percentage of the poor. Folate deficiency causes extensive incorporation of uracil into human DNA (4 million/cell), leading to chromosomal breaks. This mechanism is the likely cause of the increased colon cancer risk associated with low folate intake. Some evidence, and mechanistic considerations, suggest that Vitamin B12 (14% US elderly) and B6 (10% of US) deficiencies also cause high uracil and chromosome breaks.

Micronutrient deficiency may explain, in good part, why the quarter of the population that eats the fewest fruits and vegetables (five portions a day is advised) has about double the cancer rate for most types of cancer when compared to the quarter with the highest intake.

For example, 80% of American children and adolescents and 68% of adults do not eat five portions a day. Common micronutrient deficiencies are likely to damage DNA by the same mechanism as radiation and many chemicals, appear to be orders of magnitude more important, and should be compared for perspective. Remedying micronutrient deficiencies should lead to a major improvement in health and an increase in longevity at low cost.

1. Introduction
Approximately 40 micronutrients (the vitamins, essential minerals and other compounds required in small amounts for normal metabolism) are required in the human diet [1]. For each micronutrient, metabolic harmony requires an optimal intake (i.e. to give maximal life span); deficiency distorts metabolism in numerous and complicated ways many of which may lead to DNA damage.

The recommended dietary allowance (RDA) [2–4] of a micronutrient is mainly based on information on acute effects, because the optimum amount for long term health is generally not known. For many micronutrients, a sizable percentage of the population is deficient relative to the current RDA [5]. Remedying these deficiencies, which can be done at low cost, is likely to lead to a major improvement in health and an increase in longevity.

The optimum intake of a micronutrient can vary with age and genetic constitution, state of well being, and be influenced by other aspects of diet. Determining these optima, and remedying deficiencies, and in some cases excesses, will be a major public health project for the coming decades.

Long term health is also influenced by many other aspects of diet. Though this paper uses most examples from the US, the situation seems similar in many other countries. Micronutrient deficiency can mimic radiation (or chemicals) in damaging DNA by causing single-and double-strand breaks, or oxidative lesions, or both.

Chromosomal aberrations such as double strand breaks are a strong predictive factor for human cancer [6]. Those micronutrients whose deficiency mimics radiation are folic acid, B12, B6, niacin, C, E, iron, and zinc, with the laboratory evidence ranging from likely to compelling.

The percentage of the US population, for example, that is deficient (<50% of the RDA) for each of these eight micronutrients ranges from 2 to >20%, and may comprise in toto a considerable percentage of the US population (Table 1).

We have used <50% of the US RDA as a measure of low intake because these numbers are available [5]. However, the level of each micronutrient that minimizes DNA damage remains to be determined. Micronutrient deficiency is a plausible explanation for the strong epidemiological evidence that shows an association between low consumption of fruits and vegetables and cancer at most sites.

2. Dietary fruits and vegetables and cancer prevention
Greater consumption of fruits and vegetables is associated with a lower risk of degenerative diseases including cancer, cardiovascular disease, cataracts, and brain dysfunction [7].

More than 200 studies in the epidemiological literature have been reviewed and show, with great consistency, an association between low consumption of fruits and vegetables and the incidence of cancer [8–10]. The quarter of the population with the lowest dietary intake of fruits and vegetables has roughly twice the cancer rate for most types of cancer (lung, larynx, oral cavity, esophagus, stomach, colon and rectum, bladder, pancreas, cervix, and ovary [8] when compared to the quarter with the highest intake.

In a different survey, the lowest quartile of adults consumed 2.7 portions or less and the highest quartile 5.6 portions or more (Krebs–Smith, personal communication). These observations are consistent with data on the Seventh Day Adventists, who are non-smokers and mostly vegetarians, and have about half the cancer mortality rate and a longer life span, than the average American [11].

About 80% of American children and adolescents [12]: and 68% of adults [13] did not meet the intake recommended by the National Cancer Institute and the National Research Council: five servings of fruits and vegetables per day. Publicity about hundreds of minor hypothetical risks, such as that from pesticide residues in the diet [14], has contributed to a lack of perspective on disease prevention.

Half of Americans do not list fruit and vegetable consumption as a protective factor against cancer [15] and two-thirds think that for good health only two servings per day need to be consumed [16]. Fruit and vegetable consumption is lowest among the poor, for example, African-Americans in the US [13,17].

Many components of fruits and vegetables may be responsible for their protective effect; such as micronutrients, plant phenolics, and fiber. This paper argues that inadequate intake of many micronutrients, such as folic acid, Vitamin C and B6 contributes to DNA damage, cancer, and degenerative disease.

A major part of the protective effect of fruits and vegetables may be due to their micronutrient content. In addition, dietary deficiencies of micronutrients whose sources are not primarily fruits and vegetables, such as zinc, iron, niacin, Vitamin E, and Vitamin B12, also appear to contribute to DNA damage and are also common in the US population. Other micronutrients are likely to be added to this list in the coming years.

3. Folic acid
Folate deficiency, a common vitamin deficiency in people who eat few fruits and vegetables, causes chromosome breaks in human genes [18]. Approximately, 10% of the US population [19,20] are deficient at the level causing chromosome breaks in humans. In two small studies of low income (mainly African-American) elderly [21] and adolescents [22] done nearly 20 years ago about half had a folate deficiency at this level, though the issue should be reexamined.

(B.N. Ames / Mutation Research 475 (2001) 7–20 9, 10 / B.N. Ames / Mutation Research 475 (2001) 7–20)

The mechanism of chromosome breaks has now been shown to be deficient methylation of uracil to thymine, and subsequent incorporation of uracil into human DNA (4 million/cell) [18]. Uracil in DNA is excised by a repair glycosylase with the formation of a transient single-strand break in the DNA; two opposing single-strand breaks cause a double-strand chromosome break, which is difficult to repair. Both high DNA uracil levels and chromosome breaks in humans are reversed by folate administration [18]. Folate supplementation above the RDA minimized chromosome breakage in an Australian study [23].

Folate deficiency has been associated with increased risk of colon cancer [24,25], and the 15 year use of a multivitamin supplement containing folate lowered colon cancer risk by about 75% [26]. Folate and B12 deficiencies are associated with cognitive defects in humans [18] and neurotoxicity in children is caused by methotrexate, which lowers folate pools if folate is not replenished [27]. Chromosome breaks could contribute to the increased risk of cancer, and possibly cognitive defects, associated with folate deficiency in humans [18].

Folate deficiency causes increased homocysteine accumulation, which has been associated with neural tube defects in the fetus and an estimated 10% of US heart disease, both of which could be eliminated by folate supplements, food fortification, or better diets [28–34]. Homocysteine damages endothelial cells in culture and is a risk factor for arterial endothelial dysfunction in humans [35].

A polymorphism (a common, alternate, form of a gene) in the gene for methylene-tetrahydrofolate (THF) reductase, the enzyme responsible for reducing methylene-THF to methyl-THF, results in homozygotes having a decreased activity and a two-fold increase in plasma homocysteine.

Homozygotes, 5–25% of individuals depending on the ethnicity [36,37], have an increased risk of heart disease [31], stroke [29,38], and neural tube defects [37,39]. This polymorphism increases the methylene-THF pool at the expense of the methyl-THF pool, resulting in decreased DNA uracil levels and increased serum homocysteine.

The potential role in human carcinogenesis of uracil misincorporation is supported by two studies which show a two- to four-fold lower risk of colon cancer for individuals who are homozygous for the mutant alleles of methylene-THF reductase compared to controls [33,40]. Acute lymphocytic leukemia has been associated with the polymorphism which suggests folate deficiency as a major cause [41,42].

Folates were measured in seminal plasma from smokers and nonsmokers, and evaluated relationships between seminal plasma folates and both folate status and semen quality measures [43]. Total seminal plasma folate concentrations were higher than blood plasma folate. Total and 5-methyltetrahydrofolate concentrations correlated significantly with blood plasma folate and homocysteine concentrations. Seminal plasma non-methyltetrahydrofolates correlated significantly with sperm density and total sperm count suggesting importance for male reproductive function, and a likely mechanism of DNA damage as uracil incorporation into sperm DNA.

4. Vitamin B12
The main dietary source of B12 is meat. About 4% of the US population consumes below half of the RDA of Vitamin B12 [5]. About 14% of elderly Americans and about 24% of elderly Dutch have mild B12 deficiency, in part accountable by the Americans taking more vitamin supplements [44].

Vitamin B12 would be expected to cause chromosome breaks by the same mechanism as folate deficiency. Both B12 and methyl-THF are required for the methylation of homocysteine to methionine. If either folate or B12 is deficient, then homocysteine, a major risk factor for heart disease [29,30], accumulates.

When B12 is deficient, then tetrahydrofolate is trapped as methyl-THF; the methylene-THF pool, which is required for methylation of dUMP to dTMP, is consequently diminished. Therefore, B12 deficiency, like folate deficiency, should cause uracil to accumulate in DNA, and there is accumulating evidence for this (Ingersoll et al., unpublished; [45]). The two deficiencies may act synergistically.

In a study of healthy Australian elderly men [23], or young adults [46], increased chromosome breakage was associated with either low intakes of folate, or B12, or with elevated levels of homocysteine [47]. The B12 supplementation above the RDA was necessary to minimize chromosome breakage [46,47]. The B12 deficiency is known to cause neuropathy due to demyelination and loss of peripheral neurons (reviewed in [18]).

(B.N. Ames / Mutation Research 475 (2001) 7–20 11)

5. Vitamin B6
About 10% of the US population consumes less than half of the RDA (1.6 mg/day) of Vitamin B6 [5]. Vitamin B6 deficiency causes a decrease in the enzyme activity of serine hydroxymethyl transferase, the only source of the methylene group for methylene-THF [48]. If the methylene-THF pool is decreased in B6-deficiency, then uracil incorporation, with associated chromosome breaks, would be expected, and evidence for this has been found in women at a level of 32 nmol/l of Vitamin B6 in blood (0.5 mg/day intake) that were part of a previous intervention study ([49]; Ingersoll et al., unpublished).

In a case-control study of diet and cancer, Vitamin B6 intake was inversely associated with prostate cancer [50]. Vitamin B6 deficiency appears to contribute to heart disease and supplementation reduces risk [51]; levels above the RDA may be necessary to minimize risk [32].

A level of Vitamin B6 in blood below 23 nmol/l is a risk factor for stroke and atherosclerosis [52]. Diets low in Vitamin B6 are associated with brain dysfunction in children and adults [53]. Good sources of Vitamin B6 are whole grain bread and cereal, liver, bananas and green beans. A major source in the US is fortified breakfast cereal and multivitamins.

6. Vitamin C
About 15% of the population consumes less than half the RDA (60 mg/day) of ascorbate [5] which comes from dietary fruits and vegetables. The new RDAs for Vitamin C (90 mg/day for men, 75 mg/day for women and >35 mg for smokers) will make this percentage even higher.

There is a large literature on supplementation studies with Vitamin C in humans using biomarkers of oxidative damage to DNA, lipids (lipid oxidation releases mutagenic aldehydes), and protein. Though there are positive and negative studies, if the fact that the blood cell saturation occurs at about 100 mg/day [54,55] is taken into consideration, then the evidence suggests that this level minimizes DNA damage [56–59].

Cataracts appear to be due to oxidation of lens protein, and antioxidants, such as Vitamin C and E and carotenoids, appear to protect against cataracts and macular degeneration of the eye in rodents and humans [60–62]. The use of Vitamin C supplements for 10 years or more reduced lens opacities by about 80% [63].

Spontaneous oxidative damage in the DNA of an old rat is about 66,000 adducts per diploid cell [64,65], and unlike uracil misincorporation, is likely to be equally frequent on both strands. Glycosylase repair of oxidative adducts also results in transient single-strand breaks in DNA.

Therefore, increased oxidative damage from low Vitamin C intake, chronic inflammation, smoking, or radiation, together with elevated levels of uracil in DNA, would be expected to lead to more double-strand (chromosome) breaks in individuals who are deficient in both folate and antioxidants. There is some evidence for this synergy [66–68], which may be important because 10–15% of men in the US had serum ascorbate levels close to the scurvy threshold [5,69].

Some studies suggest that Vitamin C protects against cancer, which would be plausible based on the mechanistic data, though other studies show no effect, the variability of tissue saturation again is critical. A significant protective effect was observed for renal cancer in non-smokers, though not in smokers [70].

In a review of nutrition and pancreatic cancer, fruit and vegetable intake and Vitamin C were protective, though it is difficult to rule out that Vitamin C is a surrogate for some other compounds in fruits and vegetables [71].

Both experimental and epidemiological data suggest that Vitamin C protects against stomach cancer [72], a result that is plausible because of the role of oxidative damage from inflammation by Helicobacter pylori infection, which is the main risk factor for stomach cancer. The role of Vitamin C in inhibiting oral cancer has recently been reviewed [73].

Vitamin C improves endothelial dysfunction, an early stage of atherosclerosis, in heavy smokers [74]. Vitamin C supplementation was associated with a reduction in overall mortality and in cardiovascular disease in a follow up of the NHANES I study [75].

The effect of smoking on blood plasma antioxidant status was investigated by measuring ascorbic acid, a-tocopherol, g-tocopherol, b-carotene and lycopene and, subsequently, tested the effect of a 3-month dietary supplementation with a moderate dose vitamin cocktail [76]. Only ascorbic acid was significantly depleted by smoking per se (P < 0:01). Following the 3-month supplementation period, ascorbic acid was efficiently repleted in smokers (P < 0:001). Plasma a-tocopherol and the ratio of a- to g-tocopherol increased significantly in both supplemented groups (P <0:05).

(12 B.N. Ames / Mutation Research 475 (2001) 7–20)

The data suggests that previous reports of lower levels of plasma Vitamin E and carotenoids in smokers compared to non-smokers may primarily have been caused by differences in dietary habits between study groups. Plasma ascorbic acid is thus depleted by smoking and repleted by moderate supplementation.

Men with low consumption of antioxidants, or who smoke, oxidize the DNA of their sperm as well as their somatic DNA. When the level of dietary Vitamin C is insufficient to maintain seminal fluid Vitamin C, the oxidative lesions in sperm DNA are more than doubled [57,77]. Oxidative lesions in sperm DNA are higher in smokers than non-smokers [78].

Smoking is a severe oxidative stress, and the nitrogen oxides (NOx ) in cigarette smoke depletes antioxidants [76,79]. Thus, smokers must ingest much more Vitamin C than non-smokers to achieve the same level in

blood, but they rarely do. Inadequate Vitamin C levels are more common among the poor and smokers. Smokers also have more chromosomal abnormalities in their sperm than non-smokers [80].

Germ line mutations, and their associated cancer and genetic abnormalities, are predominately of paternal origin [81]. Smoking by fathers, therefore, may plausibly increase the risk of childhood cancer and birth defects, a thesis supported by epidemiological evidence [77,79].

The evidence on smoking fathers’ offspring having an increased rate of childhood cancer is becoming more persuasive [82–85]. A new epidemiological study from China makes the case stronger; acute lymphocytic leukemia, lymphoma, and brain cancer are each increased three- to four-fold in offspring of male smokers [82].

The studies on paternal smoking and childhood cancer did not examine the effect of diet. It seems likely, given the above evidence, that the cancer risk to offspring of male smokers would be higher when dietary antioxidant intake is low. Maternal use of multivitamins lowers the risk of childhood cancer in offspring [86].

In one study, the maternal use of vitamins throughout the pregnancy lowered the risk of brain tumors in the offspring by about half [87]. In a study of children with childhood cancer, serum levels of b-carotene, Vitamin E, and zinc were significantly lower than controls [88]. Thus, a multivitamin supplement (or a better diet) for both parents might markedly lower childhood cancer. In addition, several studies suggest an increased rate of birth defects in offspring of smoking fathers (reviewed in [77,79]).

Diets deficient in fruits and vegetables are commonly low in folate, antioxidants, (e.g. Vitamin C) and many other micronutrients, and it seems plausible that the higher cancer rates associated with consuming de- ficient diets are due, in good part, to increased DNA damage [8,18,89].

7. Vitamin E
Vitamin E, the major fat-soluble antioxidant, is consumed primarily from dietary vegetable oils and nuts. The RDA is 10 mg/day for men and 8 mg/day for women. About 20% of the population consumes less than half of the RDA [5]. Evidence is accumulating that the optimum intake may be higher, as discussed below.

Studies on Vitamin E supplementation have all been done with a-tocopherol, but g-tocopherol, the main form in the US diet, has a different function than a-tocopherol, and the two complement each other [90]. g-Tocopherol is a powerful nucleophile, and thus, traps electrophilic mutagens that reach the membrane.

In the soluble part of the cell, glutathione acts as both an antioxidant and a nucleophile. In the membrane, a-tocopherol is the antioxidant and g-tocopherol (or lycopene) can act as a nucleophile. An important electrophilic mutagen destroyed by g-tocopherol is NOx . g-Tocopherol reacts with NOx to form nitro-g-tocopherol, thus, protecting lipids, DNA, and protein [90–92]. g-Tocopherol is also an anti-inflammatory agent [93].

People taking Vitamin E supplements (200 IU/day) appear to lower their risk for colon cancer [94,95] and evidence suggests a marked protective effect of a supplement (50 IU/day) on prostate cancer [96,97]. Vitamin E appears to protect against brain dysfunction [98,99] and deficiency leads to various neuropathologies [100].

Vitamin E supplements (100–400 IU), also reduced the risk of coronary heart disease by about 40% [101–106] as well as mortality from all causes [103]. The role of oxidants and the protective role of antioxidants in heart disease have recently been reviewed [107,108]. Vitamin E is regenerated by Vitamin C.

In a study of a population with low levels of Vitamin C and E, (B.N. Ames / Mutation Research 475 (2001) 7–20 13) doses of Vitamin E from 70 to 560 IU lowered lipid peroxidation while a very high dose appeared to increase it [109] emphasizing that information on the toxic level, as well as the optimum level, of each micronutrient is desirable.

Both Vitamin E and selenium enhance the immune system in animals [110], and Vitamin E supplementation (200–400 units/day) enhances human immunity [111]. Vitamin E [112] or Vitamin C [113] reduced oxidative stress and malformations in offspring of diabetic rats.

8. Selenium
Selenium is important in enzymatic defenses against oxidants, and deficiency would be expected to lead to oxidative DNA damage [114]. An RDA of 70 mg/day of selenium and an upper limit of 350mg/day has been proposed [115]. The average intake in the US is about 100mg/day, though different areas of the country have different selenium levels in the soil, and the bioavailability depends on the selenium form in foods [114].

A growing body of evidence suggests that selenium plays an important role in the prevention of cancer in a variety of organs and species [116,117]. Prostate cancer incidence was reduced by two/thirds in the selenium supplemented group (200 mg/day) compared to the placebo group in a randomized, double-blind, cancer prevention trial; total cancer mortality, lung and colorectal cancer were also significantly reduced [118,119].

In a cohort study [120], men in the highest selenium quintile of intake had only 1/2 the odds ratio of prostate cancer as men in the lowest quintile. In a nested, case-control prospective study on ovarian cancer, serum selenium was associated with decreased risk [121].

In a study of post-menopausal breast cancer patients, a strong inverse relationship was observed between triiodothyronine (T3) levels and cancer (OR D 0:17; CI (95%/ D 0:08–0.36) between the highest and lowest tertiles [122]. Toenail selenium was positively associated with T3 levels in both cases and controls; the selenoenzyme iodothyronine deiodinase synthesizes T3. Prostate and breast cancer cells were about 25 times more sensitive than normal cells to selenomethionine, a major form of selenium in cells [123].

In a study of selenium intake and colorectal cancer that adjusted for possible confounders, the individuals in the lowest quartile of plasma selenium had four times the risk of colorectal adenomas compared to those in the highest quartile [124]. Selenium and glutathione peroxidase levels were found to be lowered in patients with uterine cervical carcinoma [125].

In a Chinese study, cervical cancer mortality was inversely associated with several factors, including serum selenium levels [126]. Selenoprotein-P level was inversely associated with several types of cancer [127]. Selenium deficiency causes human cells in culture to be more sensitive to two mutagens causing single strand breaks in DNA [128].

Several hypotheses have been proposed to explain the protection against carcinogenesis by supplemental selenium [114]. One of these is its protection against oxidative damage involving selenium as an essential component of the antioxidant enzyme glutathione peroxidase [129], or selenoprotein-P [130–132].

A recent review discusses the 11 selenoproteins and selenium’s role in preventing disease [133]. Excess selenium intake appears to cause oxidative damage and cancer in rodents [134]. The case for selenium supplementation is becoming stronger, though the toxicity of high selenium levels must be taken into account.

9. Niacin
The main dietary sources of niacin include meat and beans. About 2.3% of the US population consumes less than half the RDA of niacin [5]. Tryptophan from protein can also provide niacin equivalents [135]. About 15% of some populations have been reported to be severely deficient [136]. Niacin contributes to the repair of DNA-breaks by maintaining nicotinamide adenine dinucleotide levels for the poly-ADP ribose protective response to DNA damage [137–139]; deficiency compromises repair of DNA nicks and breaks, and thus, is expected to act synergistically with folate and antioxidant deficiencies in causing DNA damage and cancer [140].

10. Iron
A major dietary source of iron is meat. The United Nations Food and Agriculture Organization has estimated that the world has about two billion people 14 B.N. Ames / Mutation Research 475 (2001) 7–20 at risk for iron deficiency, mainly women and children. In the US, about 19% of women, aged 12–50, and about 7% of the population, ingest below 50% of the RDA [5]; about nine million people have been estimated to be clinically deficient [141].

Iron deficiency, or iron excess, leads to oxidative DNA damage [142,143]. Iron deficiency in children is associated with cognitive dysfunction [144,145]. Low iron intake results in anemia, immune dysfunction, and adverse pregnancy outcomes such as prematurity [145]. Excess iron appears to also lead to oxidative DNA damage in rats that is reversed by Vitamin E [146]. Increased risk of human cancer [145,147] and possibly heart disease [148–150] is associated with excess iron.

11. Zinc
Major sources of zinc are meat, eggs, nuts, and whole grains. Zinc deficiency causes a variety of health effects which have been reviewed in depth [151]. About 18% of the US population consumes less than half the RDA for zinc (12 mg women, 15 mg men) [5]. Mean daily intakes reported for poor children (5 mg), middle income children (6.3 mg) and vegetarians (6.4 mg) in the US appear insufficient [151].

Zinc is a component of over 300 proteins, over 100 DNA-binding proteins with zinc fingers, Cu/Zn superoxide dismutase, the estrogen receptor, and synaptic transmission protein [151]. Functioning of p53, a zinc protein which is mutated in half of human tumors, is disrupted on loss of zinc [152]. Mutation is being prevented by p53, which inhibits cell division and induces apoptosis in response to DNA lesions [153].

Chromosome breaks in rats have been reported with a zinc deficient diet [154]. The offspring of zinc deficient rhesus monkeys also have increased chromosome breaks [155]. The chromosome breaks might be due to increased oxidative damage [155,156], perhaps due to loss of activity of Cu/Zn superoxide dismutase or the zinc-containing DNA-repair enzyme, Fapy glycosylase, which repairs oxidized guanine [157].

Zinc deficiency has been suggested as a contributor to esophageal cancer in humans, and has been shown to cause esophageal tumors in rats in conjunction with a single low dose of a nitrosamine [158–160]. Severe zinc deficiency by itself can cause esophageal tumors in rats [160].

Zinc is known to be an essential trace element for testicular development and spermatogenesis [161]. Zinc concentrations in seminal plasma are hundreds of times greater than that in blood plasma, which suggests a specific function for this trace element in spermatogenesis and stability of spermatozoa [151].

Zinc concentrations are correlated positively with sperm cell density, and lower zinc concentrations are found in infertile men compared with fertile men [162]. Zinc deficiency leads to increased oxidative damage to testicular cell DNA (as measured by oxo8dG) and increased protein carbonyl content [163].

A considerable literature in experimental animals and humans suggests that zinc deficiency slows growth and development of the neonate. Severe deficiency in animals is teratogenic [155].

In a pair-matched, double-blind, study in Chile of preschool boys of low socio-economic status, those supplemented with 10 mg zinc/day grew significantly more rapidly than the placebo group [164]. This is consistent with earlier reports in the US and other countries on growth stimulation of poor children supplemented with zinc [151].

Zinc deficiency leads to alterations in brain development and growth [144]. Zinc deficiency in pregnant rats, at a level that does not impair the pregnancy or the growth of the pups, impairs cognitive function in adult offspring [151]. Zinc deficiency in adult rats impairs hippocampal and behavioral functions [151].

Several studies on monkeys show that maternal zinc deficiency leads to learning and behavioral disabilities in offspring [151]. Six studies in humans suggest that zinc deficiency leads to cognitive defects [151]. Several animal and human studies indicate that mild zinc deficiency impairs the immune system [151,165].

The incidence of respiratory infections in a group of institutionalized elderly was decreased by over two-fold (P _ 0:01) when they were given a supplement of zinc (20 mg) plus selenium (100 mg) in a double-blind placebo study; in other studies very high doses of zinc (100–150 mg/day) had an adverse effect on the immune system [166].

12. Conclusion
Optimizing micronutrient intake (through better diets, fortification of foods, or multivitamin-mineral pills [167]) can have a major impact on public health at low cost. (B.N. Ames / Mutation Research 475 (2001) 7–20 15) Other micronutrients are likely to be added to the list of those whose deficiency causes DNA damage in the coming years. Tuning-up human metabolism, which varies with genetic constitution and changes with age, is likely to be a major way to minimize DNA damage, improve health and prolong healthy lifespan.

This work was supported by National Foundation for Cancer Research Grant M2661, National Institutes of Health Grant AG17140, US. Department of Energy Grant DE-FG03-00ER62943, Tobacco-Related Disease Research Program Grant 7RT-0178, Wheeler Fund for the Biological Sciences at the University of California Berkeley, the Ellison Medical Foundation Grant SS-042-99 and National Institute of Environmental Health Sciences Center Grant ESO1896.



  1. P. Saltman, J. Gurin, I. Mothner, Nutrition Book, The University of California, San Diego, Little Brown & Company, Boston, 1993.
  2. Institute of Medicine, Dietary Reference Intakes: Calcium, Phosphorus, Magnesium, Vitamin D, and Fluoride, Committee on the Scientific Evaluation of Dietary Reference Intakes, Food and Nutrition Board, National Academy of Sciences, National Academy Press, Washington, DC, 1997.
  3. Institute of Medicine, Dietary Reference Intakes for Thiamin, Riboflavin, Niacin, Vitamin B6, Folate, Vitamin B12, Pantothenic Acid, Biotin, and Choline, Committee on the Scientific Evaluation of Dietary Reference Intakes, Food and Nutrition Board, National Academy of Sciences, National Academy Press, Washington, DC, 1998.
  4. National Research Council, Recommended Dietary Allowances, Subcommittee on the Tenth Edition of the RDAs, Food and Nutrition Board, Commission on Life Sciences, National Academy of Sciences, National Academy Press, Washington, DC, 1989.
  5. J.W. Wilson, C.W. Enns, J.D. Goldman, K.S. Tippett, S.J. Mickle, L.E. Cleveland, P.S. Chahil, Data Tables: Combined Results from USDA’s 1994 and 1995 Continuing Survey of Food Intakes By Individuals and 1994 and 1995 Diet and Health Knowledge Survey, USDA/ARS Food Surveys Research Group, Beltsville Human Nutrition Research Center, Riverdale, MD, 1997.
  6. S. Bonassi, L. Hagmar, U. Strömberg, A.H. Montagud, H. Tinnerberg, A. Forni, P. Heikkilä, S. Wanders, P. Wilhardt, I.-L. Hansteen, L.E. Knudsen, H. Norppa, Chromosomal aberrations in lymphocytes predict human cancer independently of exposure to carcinogens. European study group on cytogenetic biomarkers and health, Cancer Res. 60 (2000) 1619–1625.
  7. B.N. Ames, M.K. Shigenaga, T.M. Hagen, Oxidants, antioxidants, and the degenerative diseases of aging, Proc. Natl. Acad. Sci. U.S.A. 90 (1993) 7915–7922.
  8. G. Block, B. Patterson, A. Subar, Fruit, vegetables and cancer prevention: a review of the epidemiologic evidence, Nutr. Cancer 18 (1992) 1–29.
  9. K.A. Steinmetz, J.D. Potter, Vegetables, fruit and cancer prevention: a review, J. Am. Diet Assoc. 96 (1996) 1027–1039.
  10. W.C. Willett, D. Trichopoulos, Nutrition and cancer: a summary of the evidence, Cancer Causes Control 7 (1996) 178–180.
  11. P.K. Mills, W.L. Beeson, R.L. Phillips, G.E. Fraser, Cancer incidence among California Seventh-day Adventists, Am. J. Clin. Nutr. 59 (Suppl.) (1994) 1136S–1142S.
  12. S.M. Krebs-Smith, A. Cook, A.F. Subar, L. Cleveland, J. Friday, L.L. Kahle, Fruit and vegetable intakes of children and adolescents in the United States, Arch. Pediatr. Adolesc. Med. 150 (1996) 81–86.
  13. S.M. Krebs-Smith, A. Cook, A.F. Subar, L. Cleveland, J. Friday, US adults’ fruit and vegetable intakes, 1989–1991: a revised baseline for the healthy people 2000 objective, Am. J. Public Health 85 (1995) 1623–1629.
  14. B.N. Ames, L.S. Gold, Environmental pollution, pesticides, and the prevention of cancer: misconceptions, FASEB J. 11 (1997) 1041–1052.
  15. National Cancer Institute Graphic, Why eat five? J. Natl. Cancer Inst. 88 (1996) 1314.
  16. S. Krebs-Smith, J. Heimendinger, B. Patterson, A. Subar, R. Kessler, E. Pivonka, Psychosocial factors associated with fruit and vegetables consumption, Am. J. Health Promot. 10 (1995) 98–104.
  17. B.M. Popkin, A.M. Siega-Riz, P.S. Haines, Correction and revision of conclusions — dietary trends in the United States, N. Engl. J. Med. 337 (1997) 1846–1848.
  18. B.C. Blount, M.M. Mack, C. Wehr, J. MacGregor, R. Hiatt, G. Wang, S.N. Wickramasinghe, R.B. Everson, B.N. Ames, Folate deficiency causes uracil misincorporation into human DNA and chromosome breakage: implications for cancer and neuronal damage, Proc. Natl. Acad. Sci. U.S.A. 94 (1997) 3290–3295.
  19. F.R. Senti, S.M. Pilch, Analysis of folate data from the second national health and nutrition examination survey (NHANES II), J. Nutr. 115 (1985) 1398–1402.
  20. A.F. Subar, G. Block, L.D. James, Folate intake and food sources in the US population, Am. J. Clin. Nutr. 50 (1989) 508–516.
  21. L.B. Bailey, P.A. Wagner, G.J. Christakis, P.E. Araujo, H. Appledorf, C.G. Davis, J. Masteryanni, J.S. Dinning, Folacin and iron status and hematological findings in predominately black elderly persons from urban low-income households, Am. J. Clin. Nutr. 32 (1979) 2346–2353.
  22. L.B. Bailey, P.A. Wagner, G.J. Christakis, C.G. Davis, H. Appledorf, P.E. Araujo, E. Dorsey, J.S. Dinning, Folacin and iron status and hematological findings in black and Spanish–American adolescents from urban low-income households, Am. J. Clin. Nutr. 35 (1982) 1023–1032. 16 B.N. Ames / Mutation Research 475 (2001) 7–20.
  23. M.F. Fenech, I.E. Dreosti, J.R. Rinaldi, Folate, Vitamin B12, homocysteine status and chromosome damage rate in lymphocytes of older men, Carcinogenesis 18 (1997) 1329–1336.
  24. E. Giovannucci, M.J. Stampfer, G.A. Colditz, E.B. Rimm, D. Trichopoulos, B.A. Rosner, F.E. Speizer, W.C. Willett, Folate, methionine, and alcohol intake and risk of colorectal adenoma, J. Natl. Cancer Inst. 85 (1993) 875–884.
  25. J.B. Mason, Folate and colonic carcinogenesis: searching for a mechanistic understanding, J. Nutr. Biochem. 5 (1994) 170–175.
  26. E. Giovannucci, M.J. Stampfer, G.A. Colditz, D.J. Hunger, C. Fuchs, B.A. Rosner, F.E. Speizer, W.C. Willett, Multivitamin use, folate, and colon cancer in women in the nurses’ health study, Ann. Int. Med. 129 (1998) 517–524.
  27. C.T. Quinn, J.C. Griener, T. Bottiglieri, K. Hyland, A. Farrow, B.A. Kamen, Elevation of homocysteine and excitatory amino acid neurotransmitters in the CSF of children who receive methotrexate for the treatment of cancer, J. Clin. Oncol. 15 (1997) 2800–2806.
  28. S.A.A. Beresford, C.J. Boushey, Homocysteine, folic acid, and cardiovascular disease risk, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 193–224.
  29. C.J. Boushey, S.A. Beresford, G.S. Omenn, A.G. Motulsky, A quantitative assessment of plasma homocysteine as a risk factor for vascular disease. Probable benefits of increasing folic acid intakes, J. Am. Med. Assoc. 274 (1995) 1049–1057.
  30. G.P. Oakley Jr., M.J. Adams, C.M. Dickinson, More folic acid for everyone, now, J. Nutr. 126 (1996) 751S–755S.
  31. H. Refsum, P.M. Ueland, O. Nygård, S.E. Vollset, Homocysteine and cardiovascular disease, Ann. Rev. Med. 49 (1998) 31–62.
  32. E.B. Rimm, W.C. Willett, F.B. Hu, L. Sampson, G.A. Colditz, J.E. Manson, C. Hennekens, M.J. Stampfer, Folate and Vitamin B6 from diet and supplements in relation to risk of coronary heart disease among women, J. Am. Med. Assoc. 279 (1998) 359–364.
  33. K.L. Tucker, B. Mahnken, P.W. Wilson, P. Jacques, J. Selhub, Folic acid fortification of the food supply: potential benefits and risks for the elderly population, J. Am. Med. Assoc. 276 (1996) 1879–1885.
  34. M.J. Stampfer, F.B. Hu, J.E. Manson, E.B. Rimm, W.C. Willett, Primary prevention of coronary heart disease in women through diet and lifestyle, N. Engl. J. Med. 343 (2000) 16–22.
  35. K.S. Woo, P. Chook, Y.I. Lolin, A.S.P. Cheung, L.T. Chan, Y.Y. Sun, J.E. Sanderson, C. Metreweli, D.S. Celermajer, Hyperhomocyst(e)inemia is a risk factor for arterial endothelial dysfunction in humans, Circulation 96 (1997) 2542–2544.
  36. P. Frosst, H.J. Blom, R. Milos, P. Goyette, C.A. Sheppard, R.G. Matthews, G.J. Boers, M. den Heijer, L.A. Kluijtmans, L.P. van den Heuvel, R. Rozen, A candidate genetic risk factor for vascular disease: a common mutation in methylenetetrahydrofolate reductase (letter), Nat. Genet. 10 (1995) 111–113.
  37. A.S. Whitehead, P. Gallagher, J.L. Mills, P.N. Kirke, H. Burke, A.M. Molloy, D.G. Weir, D.C. Shields, J.M. Scott, A genetic defect in 5,10 methylenetetrahydrofolate reductase in neural tube defects, QJM 88 (1995) 763–766.
  38. H.I. Morrison, D. Schaubel, M. Desmeules, D.T. Wigle, Serum folate and risk of fatal coronary heart disease (see comments), J. Am. Med. Assoc. 275 (1996) 1893–1896.
  39. D.L. Posey, M.J. Koury, J. Mulinare, M.J. Adams Jr., C.Y. Ou, Is mutated MTHFR a risk factor for neural tube defects? Lancet 347 (1996) 686–687.
  40. J. Chen, E. Giovannucci, K. Kelsey, E.B. Rimm, M.J. Stampfer, G.A. Colditz, D. Spiegelmen, W.C. Willett, D.J. Hunter, A methylenetetrahydrofolate reductase polymorphism and the risk of colorectal cancer, Cancer Res. 56 (1996) 4862–4864.
  41. C.F. Skibola, M.T. Smith, E. Kane, E. Roman, S. Rollinson, R.A. Cartwright, G. Morgan, Polymorphisms in the methylenetetrahydrofolate reductase gene are associated with susceptibility to acute leukemia in adults, Proc. Natl. Acad. Sci. U.S.A. (1999).
  42. B.N. Ames, Cancer prevention and diet: help from single nucleotide polymorphisms, Proc. Natl. Acad. Sci. U.S.A. 96 (1999) 12216–12218.
  43. L.M. Wallock, T. Tamura, C.A. Mayr, K.E. Johnston, B.N. Ames, R.A. Jacob, Low seminal plasma folate is associated with low sperm density and number in smoking and nonsmoking men, Fertil. Steril. 75 (2001) 252–259.
  44. D.Z. van Asselt, L.C. de Groot, W.A. van Staveren, H.J. Blom, R.A. Wevers, I. Biemond, W.H. Hoefnagels, Role of cobalamin intake and atrophic gastritis in mild cobalamin deficiency in older Dutch subjects, Am. J. Clin. Nutr. 68 (1998) 328–334.
  45. S.N. Wickramasinghe, S. Fida, Bone marrow cells from Vitamin B12- and folate-deficient patients misincorporate uracil into DNA, Blood 83 (1994) 1656–1661.
  46. M. Fenech, C. Aitken, J. Rinaldi, Folate, Vitamin B12, homocysteine status and DNA damage in young Australian adults, Carcinogenesis 19 (1998) 1163–1171.
  47. M. Fenech, Micronucleus frequency in human lymphocytes is related to plasma Vitamin B12 and homocysteine, Mut. Res. 428 (1999) 299–304.
  48. S.P. Stabler, D.A. Sampson, L.P. Wang, R.H. Allen, Elevations of serum cystathionine and total homocysteine in pyridoxine-, folate-, and cobalamin-deficient rats, J. Nutr. Biochem. 8 (1997) 279–289.
  49. Y.C. Huang, W. Chen, M.A. Evans, M.E. Mitchell, T.D. Shultz, Vitamin B6 requirement and status assessment of young women fed a high-protein diet with various levels of Vitamin B6, Am. J. Clin. Nutr. 67 (1998) 208–220.
  50. T.J. Key, P.B. Silcocks, G.K. Davey, P.N. Appleby, D.T. Bishop, A case-control study of diet and prostate cancer, Br. J. Cancer 76 (1997) 678–687.
  51. E. Rimm, W. Willett, J. Manson, F. Speizer, C. Hennekens, M. Stampfer, Folate and Vitamin B6 intake and risk of myocardial infarction among US women (abstract), Am. J. Epidemiol. 143 (Suppl.) (1996) S36. B.N. Ames / Mutation Research 475 (2001) 7–20 17.
  52. K. Robinson, K. Arheart, H. Refsum, L. Brattstrom, G. Boers, P. Ueland, P. Rubba, R. Palma-Reis, R. Meleady, L. Daly, J. Witteman, I. Graham, Low circulating folate and Vitamin B6 concentrations: risk factors for stroke, peripheral vascular disease, and coronary artery disease. European COMAC Group (see comments), Circulation 97 (1998) 437–443.
  53. J.E. Leklem, Vitamin B6, in: E.E. Ziegler, L.J. Filer Jr. (Eds.), Present Knowledge in Nutrition, ILSI Press, Washington, DC, 1996, pp. 174–183.
  54. A. Kallner, D. Hartmann, D. Hornig, Steady-state turnover and body pool of ascorbic acid in man, Am. J. Clin. Nutr. 32 (1979) 530–539.
  55. M. Levine, C. Conry-Cantilena, Y. Wang, R.W. Welch, P.W. Washko, K.R. Dhariwal, J.B. Park, A. Lazarev, J.F. Graumlich, J. King, L.R. Cantilena, Vitamin C pharmacokinetics in healthy volunteers: evidence for a recommended dietary allowance, Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 3704–3709.
  56. S.J. Duthie, A. Ma, M.A. Ross, A.R. Collins, Antioxidant supplementation decreases oxidative DNA damage in human lymphocytes, Cancer Res. 56 (1996) 1291–1295.
  57. C.G. Fraga, P.A. Motchnik, M.K. Shigenaga, H.J. Helbock, R.A. Jacob, B.N. Ames, Ascorbic acid protects against endogenous oxidative damage in human sperm, Proc. Natl. Acad. Sci. U.S.A. 88 (1991) 11003–11006.
  58. D. Harats, S. Chevion, M. Nahir, Y. Norman, O. Sagee, E.M. Berry, Citrus fruit supplementation reduces lipoprotein oxidation in young men ingesting a diet high in saturated fat: presumptive evidence for an interaction between Vitamins C and E in vivo, Am. J. Clin. Nutr. 67 (1998) 240–245.
  59. M.R. McCall, B. Frei, Can antioxidant vitamins materially reduce oxidative damage in humans? Free Rad. Biol. Med. 26 (1999) 1034–1053.
  60. S.E. Hankinson, M.J. Stampfer, J.M. Seddon, G.A. Colditz, B. Rosner, F.E. Speizer, W.C. Willett, Nutrient intake and cataract extraction in women: a prospective study, Br. Med. J. 305 (1992) 335–339.
  61. S. Hung, J.M. Seddon, The relationship between nutritional factors and age-related macular degeneration, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 245–265.
  62. A. Taylor, P.F. Jacques, Antioxidant status and risk for cataract, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 267–283.
  63. P.F. Jacques, A. Taylor, S.E. Hankinson, W.C. Willett, B. Mahnken, Y. Lee, K. Vaid, M. Lahav, Long-term Vitamin C supplement use and prevalence of early age-related lens opacities, Am. J. Clin. Nutr. 66 (1997) 911–916.
  64. H.J. Helbock, K.B. Beckman, M.K. Shigenaga, P. Walter, A.A. Woodall, H.C. Yeo, B.N. Ames, DNA oxidation matters: the HPLC-EC assay of 8-oxo-deoxyguanosine and 8-oxo-guanine, Proc. Natl. Acad. Sci. U.S.A. 95 (1998) 288–293.
  65. K.B. Beckman, S. Saljoughi, S. Mashiyama, B.N. Ames, a simpler, more robust method for the analysis of 8-oxoguanine in DNA, Free Rad. Biol. Med. 3–4 (2000) 357–367.
  66. J.T. MacGregor, C.M. Wehr, R.A. Hiatt, B. Peters, J.D. Tucker, R. Langlois, R.A. Jacob, R.H. Jensen, J.W. Yager, M.K. Shigenaga, B. Frei, B.P. Eynon, B.N. Ames, Spontaneous genetic damage in man: evaluation of interindividual variability, relationship among markers of damage, and influence of nutritional status, Mut. Res. 377 (1997) 125–135.
  67. L. Wallock, A. Woodall, R. Jacob, B. Ames, Nutritional status and positive relation of plasma folate to fertility indices in nonsmoking men (abstract), FASEB J. 11 (1997) A184 –1068.
  68. R.F. Branda, D.B. Blickensderfer, Folate deficiency increases genetic damage caused by alkylating agents and gamma-irradiation in Chinese hamster ovary cells, Cancer Res. 53 (1993) 5401–5408.
  69. National Center for Health Statistics, Hematological and Nutritional Biochemistry Reference Data for Persons 6 Months–74 Years of Age: United States 1976–1980., Vital and Health Statistics, US Government Printing Office, Washington, DC, 1982.
  70. A. Wolk, P. Lindblad, H.-O. Adami, Nutrition and renal cell cancer, Cancer Causes Control 7 (1996) 5–18.
  71. G.R. Howe, J.D. Burch, Nutrition and pancreatic cancer, Cancer Causes Control 7 (1996) 69–82.
  72. S. Kono, T. Hirohata, Nutrition and stomach cancer, Cancer Causes Control 7 (1996) 41–55.
  73. S.W.Y. Chan, P.C. Reade, The role of ascorbic acid in oral cancer and carcinogenesis, Oral Dis. 4 (1998) 120–129.
  74. T. Heitzer, H. Just, T. Münzel, Antioxidant Vitamin C improves endothelial dysfunction in chronic smokers, Circulation 94 (1996) 6–9.
  75. J.E. Enstrom, L.E. Kanim, M.A. Klein, Vitamin C intake and mortality among a sample of the United States population, Epidemiology 3 (1992) 194–202.
  76. J. Lykkesfeldt, S. Christen, L.M. Wallock, H.H. Chang, R.A. Jacob, B.N. Ames, Ascorbate is depleted by smoking and repleted by moderate supplementation: a study in male smokers and nonsmokers with matched dietary antioxidant intakes, Am. J. Clin. Nutr. 71 (2000) 530–536.
  77. B.N. Ames, P.A. Motchnik, C.G. Fraga, M.K. Shigenaga, T.M. Hagen, Antioxidant prevention of birth defects and cancer, in: D.R. Mattison, A. Olshan (Eds.), Male-Mediated Developmental Toxicity, Plenum Press, New York, 1994, pp. 243–259.
  78. C.G. Fraga, P.A. Motchnik, A.J. Wyrobek, D.M. Rempel, B.N. Ames, Smoking and low antioxidant levels increase oxidative damage to sperm DNA, Mut. Res. 351 (1996) 199–203.
  79. C.A. Mayr, A.A. Woodall, B.N. Ames, DNA damage to sperm from micronutrient deficiency may increase the risk of birth defects and cancer in offspring, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 2000, pp. 373–386. 18 B.N. Ames / Mutation Research 475 (2001) 7–20.
  80. A.J. Wyrobek, J. Rubes, M. Cassel, D. Moore, S. Perrault, V. Slott, D. Evenson, Z. Zudova, L. Borkovec, S. Selevan, X. Lowe, Smokers produce more aneuploid sperm than non-smokers, Am. J. Hum. Genet. 57 (1995) 737.
  81. J. Crow, How much do we know about spontaneous human mutation rates? Environ. Mol. Mutagen. 21 (1993) 122–129.
  82. B.-T. Ji, X.-O. Shu, M.S. Linet, W. Zheng, S. Wacholder, Y.- T. Gao, D.-M. Ying, F. Jin, Paternal cigarette smoking and the risk of childhood cancer among offspring of non-smoking mothers, J. Natl. Cancer Inst. 89 (1997) 238–244.
  83. T. Sorahan, R.J. Lancashire, P. Prior, I. Peck, A.M. Stewart, Childhood cancer and parental use of alcohol and tobacco, Ann. Epidemiol. 5 (1995) 354–359.
  84. T. Sorahan, R.J. Lancashire, M.A. Hulten, I. Peck, A.M. Stewart, Childhood cancer and parental use of tobacco: deaths from 1953 to 1955, Br. J. Cancer 75 (1997) 134–138.
  85. T. Sorahan, P. Prior, R.J. Lancashire, S.P. Faux, M.A. Hulten, I. Peck, A.M. Stewart, Childhood cancer and parental use of tobacco: deaths from 1971 to 1976, Br. J. Cancer 76 (1997) 1525–1531.
  86. G.R. Bunin, J.M. Cary, Diet and childhood cancer, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 17–32.
  87. S. Preston-Martin, J.M. Pogoda, B.A. Mueller, E.A. Holly, W. Lijinsky, R.L. Davis, Maternal consumption of cured meats and vitamins in relation to pediatric brain tumors, Cancer Epidemiol. Biomarkers Prev. 5 (1996) 599–605.
  88. D.J. Malvy, J. Arnaud, B. Burtschy, D. Sommelet, G. Leverger, L. Dostalova, O. Amedee-Manesme, Antioxidant micronutrients and childhood malignancy during oncological treatment, Med. Pediatr. Oncol. 29 (1997) 213–217.
  89. B.N. Ames, L.S. Gold, W.C. Willett, The causes and prevention of cancer, Proc. Natl. Acad. Sci. U.S.A. 92 (1995) 5258–5265.
  90. S. Christen, A.A. Woodall, M.K. Shigenaga, P.T. Southwell-Keely, M.W. Duncan, B.N. Ames, g-Tocopherol traps mutagenic electrophiles such as NOx and complements a-tocopherol: physiological implications, Proc. Natl. Acad. Sci. U.S.A. 94 (1997) 3217–3222.
  91. R.V. Cooney, P.J. Harwood, A.A. Franke, K. Narala, A.K. Sundstrom, P.-O. Berggren, L.J. Mordan, Products of gamma-tocopherol reaction with NO2 and their formation in rat insulinoma (RINm5F) cells, Free Rad. Biol. Med. 19 (1995) 259–269.
  92. M.K. Shigenaga, H. Lee, B. Blount, S. Christen, E.T. Shigeno, H. Yip, B.N. Ames, Inflammation and NOx – induced nitration: assay for 3-nitrotyrosine by HPLC with electrochemical detection, Proc. Natl. Acad. Sci. U.S.A. 94 (1997) 3211–3216.
  93. Q. Jiang, I. Elson-Schwab, C. Courtemanche, B.N. Ames, g-Tocopherol and its major metabolite, in contrast to a-tocopherol, inhibit cyclooxgenase activity in macrophages and epithelial cells, Proc. Natl. Acad. Sci. U.S.A. 97 (2000) 11494–11499.
  94. E. White, J.S. Shannon, R.E. Patterson, Relationship between vitamin and calcium supplement use and colon cancer, Cancer Epidemiol. Biomarkers Prev. 6 (1997) 769–774.
  95. D. Albanes, N. Malila, P.R. Taylor, J.K. Huttunen, J. Virtamo, B.K. Edwards, M. Rautalahti, A.M. Hartman, M.J. Barrett, P. Pietinen, T.J. Hartman, P. Sipponen, K. Lewin, L. Teerenhovi, P. Hietanen, J.A. Tangrea, M. Virtanen, O.P. Heinonen, Effects of supplemental alpha-tocopherol and beta-carotene on colorectal cancer: results from a controlled trial (Finland), Cancer Causes Control 11 (2000) 197–205.
  96. T.J. Hartman, D. Albanes, P. Pietinen, A.M. Hartman, M. Rautalahti, J.A. Tangrea, P.R. Taylor, The association between baseline Vitamin E, selenium, and prostate cancer in the alpha-tocopherol, beta-carotene cancer prevention study, Cancer Epidemiol. Biomarkers Prev. 7 (1998) 335–340.
  97. O.P. Heinonen, D. Albanes, J. Virtamo, P.R. Taylor, J.K. Huttunen, A.M. Hartman, J. Haapakoski, N. Malila, M. Rautalahti, S. Ripatti, H. Maenpaa, L. Teerenhovi, L. Koss, M. Virolainen, B.K. Edwards, Prostate cancer and supplementation with alpha-tocopherol and beta- carotene: incidence and mortality in a controlled trial (see comments), J. Natl. Cancer Inst. 90 (1998) 440–446.
  98. R. Lethem, M. Orrell, Antioxidants and dementia, Lancet 349 (1997) 1189–1189.
  99. M. Sano, C. Ernesto, R.G. Thomas, M.R. Klauber, K. Schafer, M. Grundman, P. Woodbury, J. Growdon, C.W. Cotman, E. Pfeiffer, L.S. Schneider, L.J. Thal, A controlled trial of selegiline, alpha-tocopherol, or both as treatment for Alzheimer’s disease. The Alzheimer’s disease cooperative study, N. Engl. J. Med. 336 (1997) 1216–1222.
  100. R.J. Sokol, Vitamin E, in: E.E. Ziegler, L.J. Filer Jr. (Eds.), Present Knowledge in Nutrition, ILSI Press, Washington, DC, 1996, pp. 130–136.
  101. J.E. Buring, J.M. Gaziano, Antioxidant vitamins and cardiovascular disease, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 171–180.
  102. L.H. Kushi, A.R. Folsom, R.J. Prineas, P.J. Mink, Y. Wu, R.M. Bostick, Dietary antioxidant vitamins and death from coronary heart disease in postmenopausal women, N. Engl. J. Med. 334 (1996) 1156–1162.
  103. K.G. Losonczy, T.B. Harris, R.J. Havlik, Vitamin E and Vitamin C supplement use and risk of all-cause and coronary heart disease mortality in older persons: the established populations for epidemiologic studies of the elderly, Am. J. Clin. Nutr. 64 (1996) 190–196.
  104. E.B. Rimm, M.J. Stampfer, A. Ascherio, E. Giovannucci, G.A. Colditz, W.C. Willett, Vitamin E consumption and the risk of coronary heart disease in men, N. Engl. J. Med. 328 (1993) 1450–1456.
  105. M.J. Stampfer, E.B. Rimm, Epidemiologic evidence for Vitamin E in prevention of cardiovascular disease, Am. J. Clin. Nutr. 62 (Suppl.) (1995) 1365S–1369S.
  106. N.G. Stephens, A. Parsons, P.M. Schofield, F. Kelly, K. Cheeseman, M.J. Mitchinson, M.J. Brown, Randomised controlled trial of Vitamin E in patients with coronary disease: Cambridge Heart Antioxidant Study (CHAOS), Lancet 347 (1996) 781–786. B.N. Ames / Mutation Research 475 (2001) 7–20 19
  107. M.N. Diaz, B. Frei, J.A. Vita, J.F. Keaney Jr., Antioxidants and atherosclerotic heart disease, N. Engl. J. Med. 337 (1997) 408–416.
  108. A.T. Diplock, Commentary. Will the Good Fairies please prove to us that Vitamin E lessens human degenerative disease? Free Rad. Res. 27 (1997) 511–532.
  109. K.M. Brown, P.C. Morrice, G.G. Duthie, Erythrocyte Vitamin E and plasma ascorbate concentrations in relation to erythrocyte peroxidation in smoker and nonsmokers: dose response to Vitamin E supplementation, Am. J. Clin. Nutr. 65 (1997) 496–502.
  110. J.M. Finch, R.J. Turner, Effects of selenium and Vitamin E on the immune responses of domestic animals, Res. Vet. Sci. 60 (1996) 97–106.
  111. S.N. Meydani, M. Meydani, J.B. Blumberg, L.S. Leka, G. Siber, R. Loszewski, C. Thompson, M.C. Pedrosa, R.D. Diamond, B.D. Stollar, Vitamin E supplementation and in vivo immune response in healthy elderly subjects: a randomized controlled trial, J. Am. Med. Assoc. 277 (1997) 1380–1386.
  112. C.M. Simán, U.J. Eriksson, Vitamin E decreases the occurrence of malformations in the offspring of diabetic rats, Diabetes 46 (1997) 1054–1061.
  113. C.M. Simán, U.J. Eriksson, Vitamin C supplementation of the maternal diet reduces the rate of malformation in the offspring of diabetic rats, Diabetologia 40 (1997) 1416–1424.
  114. O.A. Levander, R.F. Burk, Selenium, in: E.E. Ziegler, L.J. Filer (Eds.), Present Knowledge in Nutrition, ILSI Press, Washington, DC, 1996, pp. 320–328.
  115. O.A. Levander, P.D. Whanger, Deliberations and evaluations of the approaches, endpoints and paradigms for selenium and iodine dietary recommendations, J. Nutr. 126 (1996) 2427S–2434S.
  116. E. Giovannucci, Selenium and risk of prostate cancer, Lancet 352 (1998) 755–756.
  117. P.R. Harrison, J. Lanfear, L. Wu, J. Fleming, L. McGarry, L. Blower, Chemopreventive and growth inhibitory effects of selenium, Biomed. Environ. Sci. 10 (1997) 235–245.
  118. L.C. Clark, G.F. Combs Jr., B.W. Turnbull, E.H. Slate, D.K. Chalker, J. Chow, L.S. Davis, R.A. Glover, G.F. Graham, E.G. Gross, Effects of selenium supplementation for cancer prevention in patients with carcinoma of the skin: a randomized controlled trial, J. Am. Med. Assoc. 276 (1996) 1957–1963.
  119. L.C. Clark, B. Dalkin, A. Krongrad, G.F. Combs Jr., B.W. Turnbull, E.H. Slate, R. Witherington, J.H. Herlong, E. Janosko, D. Carpenter, C. Borosso, S. Falk, J. Rounder, Decreased incidence of prostate cancer with selenium supplementation: results of a double-blind cancer prevention trial, Br. J. Urol. 81 (1998) 730–734.
  120. K. Yoshizawa, W.C. Willett, S.J. Morris, M.J. Stampfer, D. Spiegelman, E.B. Rimm, E. Giovannucci, Study of prediagnostic selenium level in toenails and the risk of advanced prostate cancer, J. Natl. Cancer Inst. 90 (1998) 1219–1224.
  121. K.J. Helzlsouer, A.J. Alberg, E.P. Norkus, J.S. Morris, S.C. Hoffman, G.W. Comstock, Prospective study of serum micronutrients and ovarian cancer, J. Natl. Cancer Inst. 88 (1996) 32–37.
  122. J.J. Strain, E. Bokje, P. van’t Veer, J. Coulter, C. Stewart, H. Logan, W. Odling-Smee, R.A. Spence, K. Steele, Thyroid hormones and selenium status in breast cancer, Nutr. Cancer 27 (1997) 48–52.
  123. C. Redman, J.A. Scott, A.T. Baines, J.L. Basye, L.C. Clark, C. Calley, D. Roe, C.M. Payne, M.A. Nelson, Inhibitory effect of selenomethionine on the growth of three selected human tumor cell lines, Cancer Lett. 125 (1998) 103–110.
  124. M.W. Russo, S.C. Murray, J.I. Wurzelmann, J.T. Woosley, R.S. Sandler, Plasma selenium levels and the risk of colorectal adenomas, Nutr. Cancer 28 (1997) 125–129.
  125. V. Bhuvarahamurthy, N. Balasubramanian, S. Govindasamy, Effect of radiotherapy and chemoradiotherapy on circulating antioxidant system of human uterine cervical carcinoma, Mol. Cell Biochem. 158 (1996) 17–23.
  126. W.D. Guo, A.W. Hsing, J.Y. Li, J.S. Chen, W.H. Chow, W.J. Blot, Correlation of cervical cancer mortality with reproductive and dietary factors, and serum markers in China, Int. J. Epidemiol. 23 (1994) 1127–1132.
  127. M.E. Persson-Moschos, L. Stavenow, B. Akesson, F. Lindgarde, Selenoprotein P in plasma in relation to cancer morbidity in middle-aged Swedish men, Nutr. Cancer 36 (2000) 19–26.
  128. A.C. Huang, M.Y. Yeh, B.N. Ames, Increased sensitivity of two selenium deficient primary human cell lines to hydrogen peroxide and to N-methyl-N0-nitro-N-nitrosoguanidine but not to UV radiation (abstract), Selenium 2000, in: Proceedings of the 7th Symposium on Selenium in Biology and Medicine, 1–5 October 2000, Venice, Italy, 2000.
  129. J.T. Rotruck, A.L. Pope, H.E. Ganther, A.B. Swanson, D.G. Hafeman, W.G. Hoekstra, Selenium: biochemical role as a component of glutathione peroxidase, Science 179 (1973) 588–590.
  130. R.F. Burk, R.A. Lawrence, J.M. Lane, Liver necrosis and lipid peroxidation in the rat as the result of paraquat and diquat administration. Effect of selenium deficiency, J. Clin. Invest. 65 (1980) 1024–1031.
  131. K.E. Hill, R.F. Burk, Selenoprotein P: recent studies in rats and in humans, Biomed. Environ. Sci. 10 (1997) 198–208.
  132. J.G. Yang, K.E. Hill, R.F. Burk, Dietary selenium intake controls rat plasma selenoprotein P concentration, J. Nutr. 119 (1989) 1010–1012.
  133. D.H. Holben, A.M. Smith, The diverse role of selenium within selenoproteins: a review, J. Am. Diet Assoc. 99 (1999) 836–843.
  134. L.S. Gold, T.H. Slone, B.N. Ames, Overview and update analyses of the carcinogenic potency database, in: L.S. Gold, E. Zeiger (Eds.), Handbook of Carcinogenic Potency and Genotoxicity Databases, CRC Press, Boca Raton, FL, 1997, pp. 661–685.
  135. G.M. McCreanor, D.A. Bender, The metabolism of high intakes of tryptophan, icotinamide and nicotinic acid in the rat, Br. J. Nutr. 5 (1986) 577–586.
  136. E.L. Jacobson, Niacin deficiency and cancer in women, J. Am. Coll. Nutr. 12 (1993) 412–416. 20 B.N. Ames / Mutation Research 475 (2001) 7–20
  137. E.L. Jacobson, W.M. Shieh, A.C. Huang, Mapping the role of NAD metabolism in prevention and treatment of carcinogenesis, Mol. Cell. Biochem. 193 (1999) 69–74.
  138. J.M. Rawling, T.M. Jackson, E.R. Driscoll, J.B. Kirkland, Dietary niacin deficiency lowers tissue poly(ADP-ribose) and NADC concentrations in Fischer-344 rats, J. Nutr. 124 (1994) 1597–1603.
  139. J.Z. Zhang, S.M. Henning, M.E. Swendseid, Poly(ADPribose) polymerase activity and DNA strand breaks are affected in tissues of niacin-deficient rats, J. Nutr. 123 (1993) 1349–1355.
  140. S.M. Henning, M.E. Swendseid, W.F. Coulson, Male rats fed methyl- and folate-deficient diets with or without niacin develop hepatic carcinomas associated with decreased tissue NAD concentrations and altered poly(ADP-ribose) polymerase activity, J. Nutr. 127 (1997) 30–36.
  141. A.C. Looker, P.R. Dallman, M.D. Carroll, E.W. Gunter, C.L. Johnson, Prevalence of iron deficiency in the United States, J. Am. Med. Assoc. 277 (1997) 973–976.
  142. M.D. Knutson, P.B. Walter, B.N. Ames, F.E. Viteri, Both iron deficiency and daily iron supplements increase lipid peroxidation in rats, J. Nutr. 130 (2000) 621–628.
  143. P.B. Walter, M.D. Knutson, A. Paler-Martinez, S. Lee, Y. Xu, F.E. Viteri, B.N. Ames, Both iron deficiency and excess supplementation damage mitochondria and mitochondrial DNA, in preparation.
  144. J. Beard, Nutrient status and central nervous system function, in: E.E. Ziegler, L.J. Filer Jr. (Eds.), Present Knowledge in Nutrition, ILSI Press, Washington, DC, 1996, pp. 612–622.
  145. R. Yip, P.R. Dallman, Iron, in: E.E. Ziegler, L.J. Filer Jr. (Eds.), Present Knowledge in Nutrition, ILSI Press, Washington, DC, 1996, pp. 277–292.
  146. D. Zhang, S. Okada, Y.Y. Yu, P. Zheng, R. Yamaguchi, H. Kasai, Vitamin E inhibits apoptosis, DNA modification, and cancer incidence induced by iron-mediated peroxidation in Wistar rat kidney, Cancer Res. 57 (1997) 2410–2414.
  147. S. Toyokuni, Iron-induced carcinogenesis: the role of redox regulation, Free Rad. Biol. Med. 20 (1996) 553–566.
  148. C.T. Sempos, R.F. Gillum, A.C. Looker, Iron and heart disease, in: A. Bendich, R.J. Deckelbaum (Eds.), Preventive Nutrition: The Comprehensive Guide for Health Professionals, Humanae Press, Totowa, NJ, 1997, pp. 181–192.
  149. T.-P. Tuomainen, K. Punnonen, K. Nyysslönen, J.T. Salonen, Association between body iron stores and the risk of acute myocardial infarction in men, Circulation 97 (1998) 1461–1466.
  150. D.G. Meyers, The iron hypothesis — does iron cause atherosclerosis? Clin. Cardiol. 19 (1996) 925–929.
  151. C.T. Walsh, H.H. Sandstead, A.S. Prasad, P.M. Newberne, P.J. Fraker, Zinc: health effects and research priorities for the 1990s, Environ. Health Perspect. 102 (Suppl. 2) (1994) 5–46.
  152. N.P. Pavletich, K.A. Chambers, C.O. Pabo, The DNA-binding domain of p53 contains the four conserved regions and the major mutation hot spots, Genes Dev. 7 (1993) 2556–2564.
  153. B. Sarkar, Metal replacement in DNA-binding zinc finger proteins and its relevance to mutagenicity and carcinogenicity through free radical generation, Nutrition 11 (1995) 646–649.
  154. C.E. Castro, L.C. Kaspin, S.-S. Chen, S.G. Nolker, Zinc deficiency increases the frequency of single-strand DNA breaks in rat liver, Nutr. Res. 12 (1992) 721–736.
  155. K.L. Olin, M.K. Shigenaga, B.N. Ames, M.S. Golub, M.E. Gershwin, A.G. Hendrickx, C.L. Keen, Maternal dietary zinc influences DNA strand break and 8-hydroxy- 20-deoxyguanosine levels in infant rhesus monkey liver, Proc. Soc. Exp. Biol. Med. 203 (1993) 461–466.
  156. P.L. Oteiza, K.L. Olin, C.E. Fraga, C.L. Keen, Oxidant defense systems in testes from zinc-deficient rats (44040), Proc. Soc. Exp. Biol. Med. 213 (1996) 85–91.
  157. T.R. O’Connor, R.J. Graves, G. de Murcia, B. Castaing, J. Laval, Fpg protein of Escherichia coli is a zinc finger protein whose cysteine residues have a structural and/or functional role, J. Biol. Chem. 268 (1993) 9063–9070.
  158. L.Y.Y. Fong, J. Li, J.L. Farber, P.N. Magee, Cell proliferation and esophageal carcinogenesis in the zinc-deficient rat, Carcinogenesis 17 (1996) 1841–1848.
  159. L.Y.Y. Fong, K.-M. Lau, K. Huebner, P.N. Magee, Induction of esophageal tumors in zinc-deficient rats by single low doses of N-nitrosomethylbenzylamine (NMBA): analysis of cell proliferation, and mutations in H-ras and p53 genes, Carcinogenesis 18 (1997) 1477–1484.
  160. P.M. Newberne, S. Broitman, T.F. Schrager, Esophageal carcinogenesis in the rat: zinc deficiency, DNA methylation and alkyltransferase activity, Pathobiology 65 (1997) 253–263.
  161. M.B. Anderson, K. Lepak, V. Farinas, W.J. Geroge, Protective action of zinc against cobalt-induced testicular damage in the mouse, Reprod. Toxicol. 7 (1993) 49–54.
  162. B. Xu, S.E. Chia, C.H. Ong, Concentrations of cadmium, lead, selenium and zinc in human blood and seminal plasma, Biol. Trace Element Res. 40 (1994) 49–57.
  163. P.I. Oteiza, K.L. Olin, C.G. Fraga, C.L. Keen, Zinc deficiency causes oxidative damage to proteins, lipids and DNA in rat testes, J. Nutr. 125 (1995) 823–829.
  164. M. Ruz, C. Castillo-Duran, X. Lara, J. Codoceo, A. Rebolledo, E. Atalah, A 14-mo zinc-supplementation trial in apparently healthy Chilean preschool children, Am. J. Clin. Nutr. 66 (1997) 1406–1413.
  165. N.W. Solomons, Mild human zinc deficiency produces an imbalance between cell-mediated and humoral immunity, Nutr. Rev. 56 (1998) 27–28.
  166. M.A. Johnson, K.H. Porter, Micronutrient supplementation and infection in institutionalized elders, Nutr. Rev. 55 (1997) 400–404.
  167. G.P. Oakley Jr., Eat right and take a multivitamin (editorial; comment) (see comments), N. Engl. J. Med. 338 (1998) 1060–1061.
  168. B.N. Ames, Micronutrients prevent cancer and delay aging, Toxicol. Lett. 102/103 (1998) 5–18.
  169. B.N. Ames, Micronutrient deficiencies: a major cause of DNA damage, in: H.L. Bradlow, J. Fishman, M.P. Osborne (Eds.), Cancer Prevention: Novel Nutrient and Pharmaceutical Developments, Annals of The New York Academy of Sciences, New York, 1999, pp. 87–106.



This paper was adapted, in part, from the work by B.N. Ames [168] and [169].
Present address: Children’s Hosp ORI, 5700 M.L.K. Way Jr., Oakland, CA 94609, USA. Tel.: C1-510-450-7625; fax: C1-510-597-7128.
E-mail address: bnames(at)uclink4.berkeley.edu (B.N. Ames).
0027-5107/01/ – see front matter © 2001 Elsevier Science B.V. PII: S0027-5107(01)00070-7 8 B.N. Ames / Mutation Research 475 (2001) 7–20 © 2001 Elsevier Science B.V. All rights reserved.

Nutrient Research References

A collection of good nutrient references from before 1999.

Links to the different categories:

Antioxidants in general, Research references

January 1999

1. Abbott RA, Cox M, Markus H, Tomkins A. Diet, body size and micronutrient status in Parkinson’s disease. Eur J Clin Nutr 46:879-84, 1992.
2. Ames BN, Shigenaga MK, Hagen TM. Oxidants, antioxidants and the degenerative diseases of aging. Proc Natl Acad Sci USA 1993; 90: 7915-22.
3. Ames BN, Gold LS, Willett WC. The causes and prevention of cancer. Proc Natl Acad Sci. USA 1995; 92: 5258-65.
4. Azen SP, Qian D, Mack W, et al. Effect of supplementary antioxidant vitamin intake on carotid arterial wall intima-media thickness in a controlled clinical trial of cholesterol lowering. Circulation 94;10:2369-72, 1996.
5. Bhat KS. Nutritional status of thiamine, riboflavin and pyridoxine in cataract patients. Nutr Rep Int 363:685-92, 1987.
6. Boers GH. Hyperhomocysteinaemia: A newly recognized risk factor for vascular disease. Neth J Med 45:1:34-41, 1994.
7. Boniton-Kopp C, Coudray C, Berr C, et al. Combined effects of lipid peroxidation and antioxidant status on carotid atherosclerosis in a population aged 59-71 y: The Eva study. Am J Clin Nutr 65:121-7, 1997.
8. Brattström L. Vitamins as homocysteine-lowering agents: A mini review. Presentation at the American Institute of Nutrition Colloquium, April 13, 1995, Atlanta, Georgia; Brattström L. Vitamins as homocysteine-lowering agents. J Nutr 126:4 Suppl:1276S-1280S, 1996.
9. Calzada C, Bruckdorfer K, Rice-Evans C. The influence of antioxidant nutrients on platelet function in healthy volunteers. Atherosclerosis 128;1: 97-105, 1997.
10. Cerhan JR, Wallace RB, Folsom AR. Antioxidant intake and risk of Parkinson’s Disease in older women. Am J Epidemiol 139;11:S65, 1994.
11. De Lorgeril , Boissonnat P, Salen P, et al. The beneficial effect of dietary antioxidant supplementation on platelet aggregation and cyclosporine treatment in heart transplant recipients. Transplantation 58:193-4, 1994.
12. De Rijk MC et al. Dietary antioxidants and Parkinson disease: The Rotterdam study. Arch Neurology 54:762-5, 1997.
13. Ely J. Crary, Smyrna, Georgia, USA – quoted in Zarrow S. Keep your eyes young and sharp. Prevention March, 1985, pp. 74-80; Crary E. Antioxidant treatment of macular degeneration of the aging and macularedema in diabetic retinopathy. South Med J 80 no. 9, supple 3:38, 1987.
14. Fahn S. An open trial of high-dosage antioxidants in early Parkinson’s disease. Am J Clin Nutr 53:380S-1S, 1991.
15. Gartside PS, Glueck CJ. Relationship of dietary intake to hospital admission for coronary heart and vascular disease: The NHANES II National Probability Study. J Am Coll Nutr 12:6:5676-84, 1993.
16. Ghosh S, et al. Dietary intake and plasma levels of antioxidant vitamins in health and disease: A hospital-based case-control study. J Nutr Environ Med 5:235-42, 1995.
17. Grimes JD et al. Prevention of progression of Parkinson’s disease with antioxidative therapy. Prog Neuropsychopharmacol Biol Psychiatry 122-3:165-72, 1988.
18. Hankinson S, Stampfer M, Seddon J, et al. Nutrient intake and cataract extraction in women: A prospective study. BMJ 305: 335-9, 1992.
19. Heliovaara M, Knekt P, Aho K, et al. Serum antioxidants and risk of rheumatoid arthritis. Ann Rheum Dis 53: 51-3, 1994.
20. Hodis HN, Mack WJ, LaBree L, et al. Serial coronary angiographic evidence that antioxidant vitamin intake reduces progression of coronary artery atherosclerosis. JAMA 273;23:1849-54, 1995.
21. Ince S. Vitamin Supplements may help delay onset of AIDS. Medical Tribune September 9, p.18, 1993.
22. Jacques PF, Chylack LT Jr, McGandy RB, Hartz SC. Antioxidant status in persons with and without senile cataract. Arch Ophthalmol 106:3:337-40, 1988.
23. Jacques PF, Chylack LT Jr. Epidemiologic evidence of a role for the antioxidant vitamins and carotenoids in cataract prevention. Am J Clin Nutr 53: 352S-5S, 1991.
24. Jacques PH et al. Vitamin intake and senile cataract. J Am Coll Nutr 6:5:435, 1987.
25. Jain VK, Chandra RK. Does nutritional deficiency predispose to Acquired Immune Deficiency Syndrome (AIDS)? Nutr Res 4:537-43, 1984.
26. Jariwalla RJ. Micro-nutrient imbalance in HIV infection and AIDS: Relevance to pathogenesis and therapy. J Nutr Environ Med 5:297-306, 1995.
27. Knekt P, Reunanen A, Jarvinen R, et al. Antioxidant vitamin intake and coronary mortality in an longitudinal population study. Am JEpidemiol 139:1180-9, 1994.
28. Knekt P, Heliovaara M, Rissanan A, et al. Serum antioxidant vitamins and risk of cataract. BMJ 305:1392-4, 1992.
29. Kritchevsky SB, Shimakawa T, Tell GS, et al. Dietary antioxidants and carotid artery wall thickness: The ARIC study. Circulation 92:8:2142-50, 1995.
30. Manson JE, Stampfer MJ, Willett WC, et al. A prospective study of antioxidant vitamins and incidence of coronary heart disease in women. Abstract. J Am Coll Nutr 11:5:633, 1992.
31. Mares-Perlman J, Klein R, Klein B, et al. Relationship between age-related maculopathy and intake of vitamin and mineral supplements. Invest Ophthalmol Vis Sci 34:1133, 1993.
32. McAlindon TE, Jacques P, Zhang Y, et al. Do antioxidant micronutrients protect against the development and progression of knee osteoarthritis? Arthritis Rheum 39(4): 648-56,1996.
33. Mastroiacovo P et al. Antioxidant Vitamins and Immunodeficiency. Int J Vitam Nutr Res 66:141-5, 1996.
34. Mooradian AD et al. Selected Vitamin and Mineral in Diabetes. Diabetes Care 17; 464-79, 1994.
35. Newsome D et al. The trace element and antioxidant economy of the human macula: Can dietary supplementation influence the course of macular degeneration? J Am Coll Nutr 10:5:536, 1991.
36. Olson RJ. Supplemental dietary antioxidant vitamins and minerals in patients with macular degeneration. J Am Coll Nutr 10:5:550, 1991.
37. Olszewski AJ et al. Reduction of plasma lipid and homocysteine levels by pyridoxine, folate, cobalamin, choline, riboflavin, and troxerutin in atherosclerosis. Atherosclerosis 75:1:1-6, 1989.
38. Pories WJ, Henzel JH, Hennessen JA. Proc U of Missouri First Annual Conference on Trace Substances in the Environment and Health 1968:114, 1967.
39. Rath M, Pauling L. A unified theory of human cardiovascular disease leading the way to the abolition of this disease as a cause for human mortality. J Orthomol Med 7:1:5-15, 1992.
40. Singh R, Niaz M, Bishnol J, et al. Diet, antioxidant vitamins, oxidative stress, and risk of coronary artery disease. Acta Cardiol 49:5:453-67, 1994.
41. Singh RB, Ghosh S, Niaz MA, et al. Dietary intake, plasma levels of antioxidant vitamins and oxidative stress in relation to coronary artery disease in elderly subjects. Am J Cardiol 76:1233-8, 1995.
42. Singh RB, Niaz MA, Ghosh S, et al. Dietary intake and plasma levels of antioxidant vitamins in health and disease: A hospital-based case-control study. J Nutr Environ Med 5:235-42, 1995.
43. Singal PK, Kapur N, Dhillon KS et al. Role of free radicals in catecholamine induced cardiomyopathy. Can J Physiol Pharmacol 60:1390, 1982.
44. Snodderly DM. Evidence for protection against age-related macular degeneration by carotenoids and antioxidant vitamins. Am J Clin Nutr 62:suppl:1448S-61S, 1995.
45. Steinberg D.Antioxidants in the prevention of human Artherosclerosis. Circulation 85; 6; 2338-2344, 1992.
46. Tang A, Graham N, Kirby A, et al. Dietary micronutrient intake and the risk of progression to Acquired Immunodeficiency Syndrome (AIDS) in Human Immunodeficiency Virus Type 1 (HIV-1) infected homosexual men. Am J Epidemiol 138: 937-51, 1993.
47. Tang AM, Graham NM, Saah AJ. Effects of Micronutrient Intake on Survival in Human immunodeficiency virus type 1 Infection. Am J Epidemiol 143:1244-56, 1996.
48. Tavani A et al. Food and nutrient intake and risk of cataract. Ann Epidemiol 6:41-6, 1996.
49. Tolonen, Matti: Fria radikaler och antioxidantia i biologi och medicin. Vitaminers og spormineralers betydning for velfærdssygdomme. (Significance of vitamins and traceminerals in disease), Rigshospitalets Symposium, Pharma Nord Research, 1988 (in Danish/Swedish).
50. Vitale S, West S, Hallfrish H, et al. Plasma antioxidants and risk of cortical and nuclear cataract. Epidemiology 4:195-203, 1993.
Ward RJ et al. Reduced antioxidant status in patients with chronic alcoholic myopathy. Biochem Soc Trans 16:581, 1988.
51. Ward RJ, Peters TJ. The antioxidant status of patients with either alcohol-induced liver damage or myopathy. Alcohol Alcohol 27;4:359-65, 1992.
52. Weisberger JH. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and carotenoids. Am J Clin Nutr 1995; 53: 226s.
53. West S, Vitale S, Hallfrisch J, et al. Are antioxidants or supplements protective for age-related macular degeneration? Arch Ophthalmol 112:222-7, 1994.
54. Woodside J et al. The effects of vitamin supplementation on cardiovascular risk. J Inherit Metabol Dis 19:Suppl 1:26/P51, 1996.

– See also betacarotene, Vitamin B6, Vitamin C, Vitamin E, Coenzyme Q10, Selenium and Zinc.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin A, Betacarotene, Research references

January 1999

1. Band P.R. et al. Treatment of Benign Breast Disease with Vitamin A. Prev Med 549-54, 1984.
2. Bendich A. Symposium conclusions: Biological actions of carotenoids. J Nutr 119: 112-5, 1989.
3. Bichler KH et al. Influence of vitamin A deficiency on the excretion of uromucoid and other substances in the urine of rats. Clin Nephrol 20:32-9, 1983.
4. Branowitz SA, Starrett B, Brookner AR. Carotene deficiency in HIV patients. AIDS 10; (1):115, 1996.
5. Burton GW, Ingold KU. Beta-carotene. An unusual type of lipid antioxidant. Scicnce 224: 569-573, 1984.
6. Cheraskin E, Ringsdorf WM, Medford FH. The ‘ideal’ daily vitamin A intake. Int J Vit Nutr Res 46: 11-13, 1976.
7. Chole Q. Vitamin A in the cochlea. Arch Otorhinolaryngol 124:379-82, 1978.
8. Comstock GW et al. Serum concentrations of alpha-tocopherol, beta-carotene, and retinol preceding the diagnosis of rheumatoid arthritis and systemic lupus erythematosus. Ann Rheum Dis 56: 323-5, 1997.
9. Connett JE, Kuller KH, Kjelsberg MO et al. Relationship between carotenoids and cancer. Cancer 64: 126-134, 1989.
10. Coutsoudis A, Bobat R, Coovadia H. The effects of vitamin-A supplementation on the morbidity of children born to HIV-infected women. Am J Public Health 85; (8):1076-81, 1995.
11. Delmas-Beauvieux M-C, Peuchant E, Couchouron A, et al. The enzymatic antioxidant system in blood and glutathione status in human immunodeficiency virus (HIV)-infected patients: Effects of supplementation with selenium or carotene. Am J Clin Nutr 64: 101-7, 1996.
12. DiMascio P, Murphy ME, Sies H. Antioxidant defense systems the role of carotenoids, tocopherols and thiols. Am J Clin Nutr 53: 194S-200S, 1991.
13. Eldred GE. Vitamins A and E in RPE lipofuscin formation and implications for age-related macular degeneration. Prog Clin Biol Res 314: 113-29, 1989.
14. Ferreira R et al. Antioxidant action of vitamins A and E in patients submitted to coronary bypass surgery. Vasc Surg 25: 191-195, 1991.
15. Gaby SK, Singh VN. Vitamin intake and health: A scientific review. New York: Marcel Dekker. p 29-57, 1991.
16. Gershoff SN, McGandy RB. The effects of vitamin A-deficient diets containing lactose in producing bladder calculi and tumors in rats. Am J Clin Nutr 34: 483, 1981.
17. Goodman DS. Vitamin A and retinoids in heath and disease. N Eng J Med 310: 1023-1031, 1984.
18. Hayes KC. Retinal degeneration in monkeys induced by deficiencies of vitamin E or A. Invest Ophthalmol Vis Sci 13:7:499-510, 1974.
19. Honkanen V et al. Vitamins A and E, retinol binding protein and zinc in rheumatoid arthritis. Clin Exp Rheumatol 7: 465-9, 1989.
20. Honkanen VEA et al. Serum cholesterol and vitamins A and E in juvenile chronic arthritis. Clin Exp Rheumatol 8: 187-91, 1990.
21. Liao CH, Erdman JW, Johnston PV. Dietary vitamin A deficiency and the immune system in a murine model of systemic lupus erythematosus. Nutr Res 16: 279-92, 1996.
22. Lohle E. The influence of chronic vitamin A deficiency on human and animal ears. Arch Otorhinolaryngol 234:167-73, 1982.
23. Mckeown LA. Beta carotene lifts CD4 counts. Study reported in Medical Tribune Feb. 25, p. 1. 1993.
24. Mobarhan S, Bowen P, Andersen B et al. Effects of beta-carotene repletion on beta-carotene absorption, lipid peroxidation, and neutrophil superoxide formation in young men. Nutr Cancer 14: 195-206, 1990.
25. Newbold PCH. Beta-carotene in the treatment of discoid lupus erythematosus. Br J Dermatol 95: 100-1, 1976.
26. Olson JA. Vitamin A. In: Present knowledge in nutrition. 7th edn. Washington DC: Intemational Life Sciences Press. p 109-119, 1996.
27. Oson JA. Provitamin A function of carotenoids. Thc conversion of B-carotene to Vitamin A. J Nutr 119: 105-108, 1989.
28. Paganini-Hill A, Chao A, Ross RK et al. Vitamin A, beta carotene and the risk of cancer. A prospective study. J Natl Cancer Inst 79: 443-448, 1987.
29. Palgi A. Association between dietary changes and morality rates; Israel 1949 to 1977; a trend-free regression model. Am J Clin Nutr 34: 1569-1583, 1981.
30. Prince MR, Frisoli JK. Beta-carotene accumulation in serum and skin. Am J Clin Nutr 1993; 57: 175-181, 1993.
31. Pryor WA. The antioxidant nutrients and disease prevention what do we know and what do we need to find out? Am J Clin Nutr 53: 391S-393S, 1991.
32. Riemersma RA, Wood DA, MacIntyre CCH, et al. Risk of angina pectoris and plasma concentrations of vitamins A, C and E and carotene. Lancet 337:1-5, 1991.
33. Romeo G. The therapeutic effect of vitamins A and E in neurosensory hearing loss. Acta Vitaminol Enzymol 7 Suppl:85-92, 1985.
34. Sahyoun NR et al. Carotenoids, vitamins C and E, and mortality in an elderly population. Am J Epidemiol 144:5:501-11, 1996.
35. Sappey C et al. Vitamin, Trace element and Peroxide status in HIV seropositive patients: Asymptomatic patients present a severe Carotene Deficiency. Clin Clim Acta 230:35-42, 1994.
36. Semba R, Graham N, Caiaffa W, et al. Increased Mortality associated with Vitamin A Deficiency during Human Immunodeficiency Virus type 1 infection. Arch Intern Med 153:2149-54, 1993.
37. Semba RD, Park S, Royal W, Griffin DE. Vitamin A deficiency and T-cell subpopulation in HIV-infected adults. Nutr Res 16:915-23, 1996.
38. Sommer A., West, KP. The duration of the effect of vitamin A supplementation. Am J Public Health 1997; 87: 467, 1997.
39. Salonen JT. Risk of cancer in relation to serum concentrations of selenium and vitamins A and E: matched case-control analysis of prospective data. Br Mcd J 1985, 290: 417-420, 1985.
40. Schauss AG. Beta-carotene and the incidence of lung cancer in Finnish male smokers. A critique. Q Rev Natural Med 191-195, 1994.
41. Seddon JM, Ajani UA, Sperduto RD, et al. Dietary carotenoids, vitamins A, C, and E, and advanced age-related macular degeneration. JAMA 272:1413-20, 1994.
42. Stahelin HB, Gey KB, Eichholzer M et al. Beta-carotene and cancer prevention. The Basel Study. Am J Clin Nutr 53: 265S-269S, 1991.
43. Street DA et al. Serum antioxidants and myocardial infarction: Are low levels of carotenoids and alpha-tocopherol risk factors for myocardial infarction? Circulation 90;3:1154-61, 1994.
44. Tang AM et al. Association between Serum Vitamin A and E levels and HIV-1 disease progression. AIDS 11:613-20, 1997.
45. Underwood BA. Was the ‘anti-infective’ vitamin misnamed? Nutr Rev 52: 140-143, 1994.
46. Vitale S et al. Plasma vitamin C, E and beta carotene levels and risk of cataract. Invest Ophthalmol Vis Sci 32:723, 1991.
47. Weisburger J. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and carotenoids. Am J Clin Nutr 53: S226-237, 1991.
48. Weisberger JH. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and carotenoids. Am J Clin Nutr 1995; 53: 226s.
49. Werler MA, Lammer EJ, Mitchell AA. Teratogenicity of high vitamin A intake. Letter. N Eng J Med 334: 1195, 1995.
50. White WS, Kim Cl, Kalkwarf HJ et al. Ultraviolet light-induced reduction in plasma carotenoid levels. Am J Clin Nutr 47: 879-883, 1988.
51. Willette W. Nutritional epidemiology. New York: Oxford University Press. p 292-310, 1990.
52. Ziegler RG. Vegetables, fruits and carotenoids and the risk of cancer. Am J Clin Nutr 53: 251S-259S, 1991.

Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B1, Research references

January 1999

1. Allison JR. The relation of hydrochloric acid and vitamin B complex deficiency in certain skin diseases. South Med J 38: 235-241, 1945.
2. Bendich A, Cohen M. Nutrition and Immunology. New York: Alan R. Liss. p 101-123, 1988.
3. Butterworth RF et al. Thiamin deficiency in AIDS. Lancet 338:1086, 1991.
4. Cheraskin E, Ringsdorf WM Jr, Setyaadmadja AT et al. Thiamin consumption and cardiovascular complaints. J Am Geriatrics Soc 15: 1074-1079, 1967.
5. Cheraskin E, Ringsdorf WM Jr, Setyaadmadja AT et al. Carbohydrate consumption and cardiovascular complaints. Angiology 18: 224-230, 1967.
6. Cheraskin E, Ringsdorf WM Jr, Medford FH et al. The ‘ideal’ daily vitamin B, intake. J Oral Med 33: 77-79, 1978.
7. Cheraskin E, Ringsdorf WM Jr. How much refined carbohydrate should we eat? Am Lab 6: 31-35, 1974.
8. Cummings F, Briggs M, Briggs, M. Vitamins in human biology and medicine. Boca Raton, FL: CRC Press. 1981.
9. Iber FL, Blass JP, Brin M et al. Thiamin in the elderly-relation to alcoholism and to neurological degenerative disease. Am J Clin Nutr 6: 1067-1082, 1982.
10. Lonsdale DA. Nutritionist’s guide to the clinical use of vitamin B1. Tacoma, WA: Life Sciences Press. p 1-209, 1987.
11. Lonsdale D. Red cell transketolase studies in a private practice specializing in nutritional correction. J Am Coll Nutr 7: 61-68, 1988.
12. Rindi, G. Thiamin. In: Present knowledge in nutrition. 7th edn. Washington DC: International Life Sciences Press. p 160-166, 1996.
13. Ropert R. Utilisation de la Vitamine B1 fortes doses dans le traitement de divers syndromes douloureux facio-céphaliques. Sem Hôp 4:508-9, 1958 (in French).
14. Shils ME, Young VR. Modern nutrition in health and disease. 7th edn. Philadelphia: Lea and Febiger. p 358, 1988.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B2, research references

January 1999

1. Belko AZ, Obarzanek E, Kalkwarf HJ et al. Effects of exercise on riboflavin requirements of young women. Am J Clin Nutr 37: 509-517, 1983.
2. Belko AZ, Obazanek E, Roach R et al. Effects of aerobic exercise and weight loss on riboflavin requirements of moderately obese, marginally deficient young women. Am J Clin Nutr 40: 553-561, 1984.
3. Belko AZ, Meredith MP, Kalkwarf HJ et al. Effects of exercise on riboflavin requirements. Biological validation in weight reducing women. Am J Clin Nutr 41: 270-277, 1985.
4. Bendich A, Cohen M. Nutrition and Immunology. New York: Alan R. Liss. p. 114, 1988.
5. Beutler E. Glutathione reductase. Stimulation in normal subjects by riboflavin supplementation. Science 165: 614-615, 1969.
6. Goodwin JS, Garry PJ. Relationship between megadose vitamin supplementation and immunological function in a healthy elderly population. Clin Exp Immunol 51: 647-653, 1983.
7. Joint, FAO/WHO Expert Group. Riboflavin. WHO Technical Report Series No. 362. p. 86, 1967.
8. Prchal JT, Conrad ME, Skalka HW. Association of presenile cataracts with heterozygosity for glactosaemic states and riboflavin deficiency. Lancet i: 12-143, 1978.
9. Rivlin RS. Riboflavin. In: Ziegler EE, Filer LJ Jr, eds. Present knowledge in nutrition. 7th edn. Washington DC: International Life Sciences Press. p 167-173, 1996.
10. Salim-Hanna M, Edwards AM, Silva E. A photo-induced adduct between a vitamin and an essential amino acid: binding of riboflavin to tryptophan. Int J Vit Nutr Res 57: 155-159, 1987.
11. Schoenen J, Jacquay J, Lenaerts M. Effectiveness of high-dose riboflavin in migraine prophylaxis: a randomized controlled trial. Neurology 50: 466-470, 1998.
12. Schoenen J, Lenaerts M, Bastings E. High-dosc riboflavin as a prophylactic treatment of migraine: results of an open pilot study. Cephalalgia 14: 328-329, 1994.
13. Skalka HW, Prchal JT. Cataracts and riboflavin deficiency. Am J Clin Nutr 34: 861-863, 1981.
14. Webster RP, Gawde MD, Bhattacharya RK. Modulation of carcinogen-induced DNA damage and repair enzyme activity by dietary riboflavin. Cancer Lett 98: 129-135, 1996.
15. Weisburger J. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and cartenoids. Am J Clin Nutr 53: S226-227, 1991.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B3, Research references

January 1999

1. Alhadeff L, Gualtieri GT, Lipton M. Toxic effects of water-soluble vitamins. Nutr Rev 42: 33-40, 1984.
2. Bendich A, Cohen M. Nutrition and immunology. New York: Alan R. Liss. p 114-115, 1988.
3. Carlson LA, Hamsten A, and Asplund A. Pronounced Lowering of Serum Levels of Lipoprotein Lp(a) in Hyperlipidaemic Subjects Treated with Nicotinic Acid, J Intern Med 226, 271-6, 1989.
4. Cheraskin E, Ringsdorf WM Jr, Medford FH. The ‘ideal’ daily niacin intake. Int J Vit Nutr Res 46: 58-60, 1976.
5. Cleary JP. Vitamin B3 in the Treatment of Diabetes Mellitus: Case Reports and Review of the Literature. J Nutr Med 1: 217-25, 1990.
6. DiPalma JR and Thayer WS, Use of Niacin as a Drug, Ann Rev Nutr 11; 169-87, 1991.
7. Einstein N, Baker A, Galper J et al. Jaundice due to nicotinic acid therapy. Am J Digest Dis 20: 282-286, 1975.
8. El-Enein AMA et al., The Role of Nicotinic Acid and Inositol Hexaniacinate as Anticholesterolemic and Antilipemic Agents, Nutr Rep Intl 28, 899-911, 1983.
9. Figge HL, Figge J, Souney PF et al. Nicotinic acid. A review of its clinical use in the treatment of lipid disorders. Pharmacotherapy 8: 287-294, 1988.
10. Gaby SK. Vitamin intake and health: A scientific review. New York: Marcel Dekker. p 189-192, 1991.
11. Goldsmith GA, Miller ON, Unglaub WG. Efficiency of tryptophan as a niacin precursor in man. J Nutr 73: 172-176, 1961.
12. Grundy SM. Drug therapy in dyslipidemia. Scand J Clin Lab Invest 50: 63-72, 1990.
13. Illingworth DR et al., Comparative Effects of Lovastatin and Niacin in Primary Hypercholesterolemia, Arch Intern Med 154, 1586-95, 1994.
14. Jacob, RA, Swendseid, ME. Niacin. In: Present knowledge in nutrition. 7th edn. Washington DC: International Life Sciences Press. p 184-190, 1996.
15. Jonas WB et al. The effect of niacinamide on osteoarthritis: a pilot study. Inflamm Res 45: 330-4, 1996.
16. Lefavi R. Lipid-lowering effects of a dietary nicotinic acid-chromium III complex in male athletes. FASEB J 5;6:A1645, 1991.
17. Lefavi R et al. Lipid-lowering effect of a dietary chromium III-nicotinic acid complex in male athletes. Nutr Res 13:239-49, 1993.
18. Luria MH. Effect of low-dose niacin on high-density lipoprotein cholesterol and total cholesterol/high density lipoprotein cholesterol ratio. Arch Intern Med 148: 2493-5, 1988.
19. Mandrup Paulsen T. Nicitinamide in the Prevention of Insulin Dependent Diabetes Mellitus. Diabetes Metabol Rev 9: 295-309, 1993.
20. McKenney JM et al., A Comparison of the Efficacy and Toxic Effects of Sustained- vs Immediate-Release Niacin in Hypercholesterolemic Patients, JAMA 271, 672-7, 1994.
21. O’Hara J, Jolly PN, Nicol CG. The therapeutic efficacy of inositol nicotinate (I lexopal) in intermittent claudication of a controlled trial. Br J Clin Practice 42: 377-383, 1988.
22. Patterson JI, Brown RR, Lindswiler H et al. Exertion of tryptophan-niacin metabolites by young men. Effects of tryptophan, leucine, and vitamin B6 intakes. Am J Clin Nutr 33: 2157-2167, 1980.
23. Pocoit F, Reimers JI and Andersen HU. Nicotinamide: Biological Actions and Therapeutic Potential in Diabetes Prevention. Diabetologia 36: 574-76, 1993.
24. Pozzilli P et al. The Potential Role of Nicotinamide in the Secondary Prevention of IDDM. Diabetes Metabol Rev 9: 219-30, 1993.
25. Salem and Abdel M. The Role of Nicotinic Acid and Inositol Hexaniacinate as Anticholesterolemic and Antilipemic Agents. Nutr Rep Intl 28: 899-911, 1983.
26. Urberg M and Zemel MB. Evidence for Synergism Between Chromium and Nicotinic Acid in the Control of Glucose Tolerance in Elderly Humans. Metabolism 36: 896-99, 1987.
27. Urberg M et al. Hypocholesterolemic effect of nicotinic acid and chromium supplementation. J Fam Pract 27;6:603-6, 1988.
28. Vega GL and Crundy SM. Lipoprotein Responses to Treatment with Lovastatin, Gemfibrozil, and Nicotinic Acid in Normolipidemic Patients with Hypoalphalipoproteinemia, Arch Intern Med 154: 73-82, 1994.
29. Welsh AL and Ede M, Inositol Hexanicotinate for Improved Nicotinic Acid Therapy, Int Record Med 174: 9-15, 1961.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B5, Research references

January 1999

1. Barton-Wright EC, Elliott WA. The Pantothenic Acid Metabolism of Rheumatoid Arthritis. Lancet ii: 862-3, 1963.
2. Fry PC et al. Metabolic response to a pantothenic acid deficient diet in humans. J Nutr Sci Vitaminol 22: 339-46, 1976.
3. Welsh AL. Lupus erythematosus: Treatment by combined use of massive amounts of pantothenic acid and vitamin E. Arch Dermatol Syphilol 70: 181-98, 1954.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B6, Research references

January 1999

1. Annand JC. Pyridoxine and magnesium in the treatment of shock. Lancet ii: 340-1, 1957.
2. Antopol W, Schotland CE. The use of vitamin B6 in pseudohypertrophic muscular dystrophy. JAMA, March 23, pp. 1058-9, 1940.
3. Barr W, Pyridoxine Supplements in the Premenstrual Syndrome, Practitioner 228: 425-7, 1984.
4. Bassler KH. Megavitamin therapy with pyridoxine. Int J Vit Nutr Res 58: 105-118, 1988.
5. Berman MK, et al. Vitamin B6 in Premenstrual Syndrom, J Am Diet Assoc 90 : 859-61, 1990.
6. Brattström L et al. Impaired homocysteine metabolism in early-onset cerebral and peripheral occlusive arterial disease. Effects of pyridoxine and folic acid treatment. Atherosclerosis 81:1:51-60, 1990.
7. Brattström L, Stavenow L, Galvard H, et al. Pyridoxine reduces cholesterol and low-density-lipoprotein and increases antithrombin III activity in 80- year-old men with low plasma pyridoxal 5-phosphate. Scand J Clin Lab Invest 50:8:873-7, 1990.
8. Bum MK et al. Association of Vitamin B6 Status with Parameters of Immune Function in Early HIV-1 Infection. J AIDS 4:122-32, 1991.
9. Caby SK. Vitamin intake and health: A scientific review. New York: Marcel Dekker. p 163- 74, 1991.
10. Cohen M, Bendich A. Safety of pyridoxine – a review of human and animal studies. Toxicol Letters 34: 129-139, 1986.
11. Curhan GC, Willett WC, Rimm EB, Stampfer MJ. A prospective study of the intake of vitamins C and B6, and the risk of kidney stones in men. J Urol 155:61847-51, 1996.
12. Dalery K et al. Homocysteine and coronary artery disease in French Canadian subjects: Relation with vitamins B12, B6, pyridoxal phosphate and folate. Am J Cardiol 75:1107-11, 1995.
13. Driskell JA, Wesley RL, Hess IE. Effectiveness of pyridoxine hydrochloride treatment on carpal tunnel syndrome patients. Nutr Rep Int 1986; 34: 103-1040, 1986.
14. Ellis JM. Vitamin B6 deficiency and rheumatism. Anabolism Winter 1985.
15. Folkers K, Morita M, McRee Jr. J. The activities of coenzyme Q10 and vitamin B6 for immune response. Biochem Biophys Res Commun 193:88-92, 1993.
16. Gaby AR. The safe use of vitamin B6 J Nutr Med 1: 153-157, 1990.
17. Gershoff SN, Prien EL. Excretion of urinary metabolites in calcium oxalate urolithiasis: Effect of tryptophan and vitamin B6 administration. Am J Clin Nutr 8:812, 1960.
18. Gershoff S, Prien EL. Effect of daily MgO and vitamin B6 administration to patients with recurring calcium oxalate kidney stones. Am J Clin Nutr 20:393-399, 1967.
19. Gibbs DA, Watts RWE. The action of pyridoxine in primary hyperoxaluria. Clin.Sci 38:277-86, 1970.
20. Gvozdova LG, Paramonova EG, Goriachenkova EV et al. The content of pyridoxal coenzymes in the blood plasma of patients with coronary atherosclerosis on a background of therapeutic diet and after supplemental intake of vitamin B6. Vop Pitan 25: 40-44, 1966.
21. Hallert C, Astrom J, Walan A. Reversal of psychopathology in adult coeliac disease with the aid of pyridoxine (vitamin B6). Scand J Gastroenterol 18; (2):299-304, 1983.
22. Harrison AR et al. Hyperoxaluria and recurrent stone formation apparently cured by short course of pyridoxine. Br Med J 282: 2097-8, 1981.
23. Jaeger P. et al. Idiopathic hyperoxaluria cured by a short course of pyridoxine B6: Further evidence for B6-deficiency. Abstract. Clin Res 34, 2:546A, 1962.
24. Jones CL et al. Pyridoxine Deficiency: A New Factor in Diabetic Neuropathy. J Am Pod Assoc 68: 646-53, 1978.
25. Kasidas GP et al. Mild but clinically significant metabolic hyperoxaluria and its response to pyridoxine, in H von G Gasser, Wahlensieck W, Eds. Pathogenese und Klinik Harnstein XI. Darmstadt, Steinkopff Verlag, 394-9, 1985.
26. Kok FJ et al. Low vitamin B6 status in patients with acute myocardial infarction. Am J Cardiol 63:513-16, 1989.
27. Kuzuya F. Arteriosclerosis in pyridoxine deficient-monkeys. Primates 2:99, 1959.
28. Kuzuya F F. Experiment on arteriosclerosis and arteriolosclerosis induced in pyridoxine-deficient monkeys and their recovery. Primates 3:77, 1962.
29. Kuzuya F. Reversibility of arteriosclerosis in pyridoxine-deficient monkeys, in G Schettler, Y Goto, Y Hata, G Klose, Eds. Atherosclerosis IV. Berlin, Springer-Verlag, 275-8, 1977.
30. Kuzuya F. Vitamin B6 and arteriosclerosis. Nagoya J Med Sci 55:1-4:1-9, 1993.
31. Lam SC et al. Investigation of possible mechanisms of pyridoxal-5-phosphate inhibition of platelet reactions. Thrombosis Res 20:633-45, 1980.
32. Leklem, JE. Vitamin B-6. In: Present knowledge in nutrition.7th edn. Washington DC: International Life Sciences Press. p 174-83, 1996.
33. Levene CI, Murray JC. The aetiological role of maternal B6 deficiency in the development of atherosclerosis. Lancet i:628, 1977.
34. Lewy A, Fox N. Clinical notes; New instruments and technics: Pyridoxine (B6) used in the treatment of vertigo. Arch Otolaryngol pp.681-3, Nov. 1947.
35. Lyon E, Borden T, Ellis J, Vermeulen C. Calcium oxalate lithiasis produced by pyridoxine deficiency and inhibition with high magnesium diets. Invest Urol 4: 133-142, 1966.
36. McCully KS. Vascular pathology of homocysteinemia: Implications for the pathogenesis of arteriosclerosis. Am J Pathol 56: 111-128, 1969.
37. Milliner D, Eickholt J, Berstrahl E, et al. Results of long-term treatment with orthophosphate and pyridoxine in patients with primary hyperoxaluria. N Engl J Med 331; 23:1553-8, 1994.
38. Mitwalli A et al. Control of hyperoxaluria with large doses of pyridoxine in patients with kidney stones. Int Urol Nephrol 20; 4:353-9, 1988.
39. Murthy MS, Farooqui S, Talwar HS, et al. Effect of pyridoxine supplementation on recurrent stone formers. Int J Clin Pharmacol Ther Toxicol 20; 9: 434-7, 1982.
40. Mushett CW, Emerson G. Arteriosclerosis in pyridoxine-deficient monkeys and dogs. Fed Proc 4:526-7, 1956.
41. Parry GJ, Bredesen DE. Sensory neuropathy with low dose pyridoxine. Neurology 1985; 35: 1466- 1468.
42. Prien EL, Gershoff S. Magnesium oxide-pyridoxine therapy for recurring calcium oxalate urinary calculi. J Urol 112:509-512, 1974.
43. Rattan VF, Sidhu H, Vaidyanathan S, et al. Effect of combined supplementation of magnesium oxide and pyridoxine in calcium-oxalate stone formers. Urol Res 22; 3:161-5, 1994.
44. Reinken L, Zieglauer H. Vitamin B6 absorption in children with acute celiac disease and in control subjects. J Nutr 108:1562, 1978.
45. Revusova V et al. The significance of oxaluria reduction by pyridoxine in prevention of calcium oxalate nephrolithiasis. Cas Lek Ces 121; 6:163-6, 1982.
46. Rinehart JF, Greenberg LD. Arteriosclerotic lesions in pyridoxine deficient monkeys. Am J Pathol 25:481-96, 1949.
47. Rinehart JF, Greenberg LD. Pathogenesis of experimental arteriosclerosis in pyridoxine deficiency with notes on similarities to human arteriosclerosis. Arch Pathol 51:12-29, 1951.
48. Rinehart JF, Greenberg LD. Vitamin B6 deficiency in the Rhesus monkey. Am J Clin Nutr 4:318-325, 1956.
49. Rhinehart JF, Greenberg LD. Vitamin B6 deficiency in the rhesus monkey, with particular reference to the occurrence of atherosclerosis, dental caries and hepatic cirrhosis. Am J Clin Nutr 4:318, 1956.
50. Roubenoff R et al. Abnormal vitamin B6 status in rheumatoid cachexia. Arthritis Rheum 1: 105-9, 1995.
51. Serofontein WJ, Ubbink JB, De Villiers LS, et al. Plasma pyridoxal-5-phosphate level as risk index for coronary artery disease. Atherosclerosis 55:3:357-61, 1985.
52. Serofontein WJ, Ubbink JB, De Villers LS, Becker PJ. Depressed plasma pyridoxal-5′-phosphate levels in tobacco-smoking men. Atherosclerosis. 59: 341-346; 1986.
53. Shor-Posner G, Feaster D, Blaney N, et al. Impact of Vitamin B6 Status on Psychological Distress in a longitudinal Study of HIV-1 infection. Int J Psychol Med 24; (3):209-22, 1994.
54. Shultz TD, Santamaria AG, Gridley DS et al. Effect of pyridoxine and pyridoxal on the in vitro growth of human malignant melanoma. Nutr Res 8: 201-207, 1988.
55. Solomon LR et al. Erythrocyte O2 Transport and Metabolism and Effects of Vitamin B6 Therapy in Type II Diabetes mellitus. Diabetes 38: 881-86, 1989.
56. Stone S. Pyridoxine and thiamine therapy in disorders of the nervous system. Dis Nerv Sys 11;5:131-8, 1950.
57. Subbarao K et al. Pyridoxal 5’-phosphate – a new physiological inhibitor of blood coagulation and platelet function. Biochem Pharmacol 28:531-4, 1979.
58. Swift ME, Shultz TD. Relationship of vitamins B6 and Bl2 to homocysteine levels: Risk for coronary heart disease. Nutr Rep Int 34: 1-14, 1986.
59. Talbott MC, Miller LT, Kerkvliet NI. Pyridoxine supplementation effect on Lymphocyte responses in elderly persons. Am J Clin Nutr 46: 659-664, 1987.
60. Thind SK et al. Role of vitamin B6 in oxalate metabolism in urolithiasis. Abstract. Am J Clin Nutr 32; 6:20, 1979.
61. Ubbink JB, Vermaak WJH, van der Merwe A, Becker PJ. Vitamin B-12, vitamin B-6, and folate nutritional status in men with hyperhomocysteinemia. Am J Clin Nutr 57:47-53, 1993.
62. Verhoef P, Stampfer MJ, Buring JE, et al. Homocysteine metabolism and risk of myocardial infarction: Relation with vitamin B6, B12, and folate. Am J Epidemiol 143:9:845-59, 1996.
63. Verrmaak WJ, Barnard HC, Potgieter GM et al. Plasma pyridoxal 5′-phosphate levels in myocardial infarction. S Afr Med J 70: 195-196, 1986.
64. Vermaak WJH et al. Vitamin B6 and coronary heart disease. Epidemiological observations and case studies. Atherosclerosis 63:235, 1987.
65. Vijayammal PL, Kurup PA. Pyridoxine and atherosclerosis: Role of pyridoxine in the metabolism of lipids and glycosaminoglycans in rats fed normal and high fat, high cholesterol diets containing 16% casein. Austral J Biol Sci 31:1:7-20, 1978.
66. Weisburger J. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidant, and carotenoids. Am J Clin Nutr 53: S226-S237, 1991.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.

Vitamin B9, Folic acid, Folate, Research references

January 1999

1. Alpert JE, Fava, M. Nutrition and depression. The role of folate. Nutr Rev 55: 145-149, 1997.
2. Bailey LB, Wagner PA, Davis CG, Dinning JS. Food frequency related to folacin status in adolescents. J Am Diet Assoc 84: 801-804, 1984.
3. Baker H, Jaslow SP, Frank O. Severe impairment of dietary folate utilization in the elderly. J Am Geriatr Soc 26: 218-221, 1978.
4. Bates CJ, Fleming M, Paul AA et al. Folate status and its relation to vitamin C in healthy elderly men and women. Age Aging 9: 241-248, 1980.
5. Bendich A, Cohen M. Nutrition and immunology. New York: Alan R. Liss. p 101-123, 1988.
6. Botez MI, Botez T, Léveillé J, et al. Neuropsychological correlates of folic acid deficiency: Facts and hypotheses, in MI Botez, EH Reynolds, Eds. Folic Acid in Neurology, Psychiatry, and Internal Medicine. New York, Raven Press, 435-461, 1979.
7. Brattstrom LE, Hultberg BL, Hardebo JE. Folic acid responsive postmenopausal homocysteinemia. Metabolism 34: 1073-1077, 1987.
8. Brattstrom LE, Israelsson B, Jeppsson JO, Hultberg BL. Folic acid – an innocuous means to reduce plasma homocysteine. Scand J Clin Lab Invest 48: 3; 215-221, 1988.
9. Brattström L et al. Impaired homocysteine metabolism in early-onset cerebral and peripheral occlusive arterial disease. Effects of pyridoxine and folic acid treatment. Atherosclerosis 81:1:51-60, 1990.
10. Briggs RM. Vitamin supplementation as a possible factor in the incidence of cleft lip/palate deformities in humans. Clin Plast Surg 1976; 3: 647-652, 1976.
11. Butterworth CE, Tamura T. Folic acid safety and toxicity. A brief review. Am J Clin Nutr 50: 353-358, 1989.
12. Center for Disease Control. Knowledge about folic acid and use of multivitamins containing folic acid among reproductive-aged women. Morbidity Mortality Weekly Report 45: 793-795, 1996.
13. Center for Disease Control. Recommendations for the use of folic acid to reduce the number of cases of spina bifida and other neural tube defects. Morbidity Mortality Weekly Report 41: RR-14, 1996.
14. Clark AJ, Mossholder S, Gates R. Folacin status in adolescent females. Am J Clin Nutr 1987; 46: 302-306.
15. Dalery K et al. Homocysteine and coronary artery disease in French Canadian subjects: Relation with vitamins B12, B6, pyridoxal phosphate and folate. Am J Cardiol 75:1107-11, 1995.
16. Deller DJ et al. Folic acid deficiency in rheumatoid arthritis. Relation of levels of serum folic acid activities to treatment with phenylbutazone. Br Med J i: 765-7, 1966.
17. Flynn MA, Irvin W, Krause G. The effect of folate and cobalamin on osteoarthritic hands. J Am Coll Nutr 13; 4: 351-6, 1994.
18. Gaby SK, Bendich A. Vitamin intake and health: A scientific review. New York: Marcel Dekker. p 175-188, 1991.
19. Gough KR et al. Folic acid deficiency in rheumatoid arthritis. Br Med J i: 212-17, 1964.
20. Halsted CH, Reisenauer AM, Romero JJ, et al. Jejunal perfusion of simple and conjugated folates in celiac sprue. J Clin Invest 59: 933-40, 1977.
21. Hjelt K, Krasilnikoff PA. The impact of gluten on haematolgical status, dietary intakes of haemopoietic nutrients and vitamin B12 and folic acid absorption in children with coeliac disease. Acta Paediatr Scand 79; (10):911-19, 1990.
22. Huber AM, Wallins LL, DeRusso P. Folate nutriture in pregnancy. J Am Diet Assoc 88: 791-814, 1988.
23. Jacques P, Bostom A, Williams R, et al. Relation between folate status, a common mutation in methylene tetrahydrofolate reductase, and plasma homocysteine concentrations. Circulation 93; 1: 7-9, 1996.
24. Kang SS, Wong PWK, Norusis M. Homocysteinemia due to folate deficiency. Metabolism 36: 458-462, 1987.
25. Laurence KM, James N, Miller MH et al. Double-blind randomized controlled trial of folate treatment before conception to prevent recurrence of neural-tube defects. Br Med J 282: 1509-1511, 1981.
26. Milunsky A, Jick H, Jick SS, Bruell CL, MacLaughlin DS, Rothman KJ, Willette W. Multivitamin/folic acid supplementation in early pregnancy reduces the pr-reviewence of neural tube defects. J Am Med Assoc 262: 2847-2852, 1989.
27. Modesto C, Castro L. Folinic acid supplementation in patients with juvenile rheumatoid arthritis treated with methotrexate. J Rheumatol 23; (2): 403-4, 1996.
28. Morgan SI, Alarcon GS, Krumdieck CL. Folic acid supplementation during methotrexate therapy: It makes sense. Editorial. J Rheumatol 20; (6): 929-30, 1993.
29. Morgan SL, Baggott JE, Vaughn WH, et al. Supplementation with folic acid during methotrexate therapy for rheumatoid arthritis: a double-blind, placebo-controlled trial. Ann Intern Med 121: 833-41, 1994.
30. Murray, MT. reviewuating the many benefits of folic acid. Am J Natural Med 3: 8-11, 1996.
31. Omer A, Mowat AG. Nature of anaemia in rheumatoid arthritis. IX. Folate metabolism in patients with rheumatoid arthritis. Ann Rheum Dis 27: 414-24, 1968.
32. Preuss HG. CRC Handbook series in nutrition and food. Section F: Nutntional disorders, vol. 1. Boca Raton, FL: CRC Press. p 61-62, 1978.
33. Selhub, J, Rosenberg, IH. Folic acid. In: Present knowledge in nutrition. 7th edn. Washington DC: International Life Sciences Press. p 206-219, 1996.
34. Smithells RW, Nevin NC, Seller MJ et al. Further experience of vitamin supplementation for prevention of neural tube defect recurrences. Lancet 1: 1027-1031, 1983.
35. Ubbink JB, Vermaak WJH, van der Merwe A, Becker PJ. Vitamin B-12, vitamin B-6, and folate nutritional status in men with hyperhomocysteinemia. Am J Clin Nutr 57:47-53, 1993.
36. Ventura A, bouquet F, Sartorelli C, et al. Coeliac disease, folic acid deficiency and epilepsy with cerebral calcifications. Acta Paeditr Scand 80; (5):559-62, 1991.
37. Verhoef P, Stampfer MJ, Buring JE, et al. Homocysteine metabolism and risk of myocardial infarction: Relation with vitamin B6, B12, and folate. Am J Epidemiol 143:9:845-59, 1996.
38. Weisburger JH. Nutritional approach to cancer prevention with emphasis on vitamins, antioxidants, and carotenoids. Am J Clin Nutr 53: S226-S237, 1991.


Joseph E. Pizzorno Jr., Michael T. Murrey & Melvyn R. Werbach.