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Investigation of folate intake and metabolism in women who have had two pregnancies complicated by neural tube defects.

OBJECT: To investigate folate intake and blood levels of folic acid and vitamin C in women with and without a history of two NTD-affected pregnancies and to measure the increase in serum folate following ingestion of orange juice. SUBJECTS: Sixteen women with a history of two NTD-affected infants and 16 controls with no such history, none of whom were either pregnant or taking vitamin supplements. The orange juice loading test was carried out on eight matched pairs. MAIN OUTCOME MEASURES: Dietary intake of folate was assessed by questionnaire and blood levels of folate and vitamin C were measured. RESULTS: There was no evidence of decreased intake of folate in subjects who had had two NTD-affected pregnancies. In controls, both serum and red cell folate showed significant correlation with dietary folate, while in subjects there was no such correlation. Subjects also had smaller increases in serum folate following an oral load than controls, although the differences were not significant. CONCLUSION: Women who have had two NTD-affected pregnancies may have defective folate metabolism. Further investigations on short term utilisation of ingested food folates in these women are required.

Adult↗

Proteinuria and plasma total homocysteine levels in chronic renal disease patients with a normal range serum creatinine: critical impact of true glomerular filtration rate.

Conflicting data have been reported concerning the independent association between proteinuria and plasma total homocysteine (tHcy) levels, particularly among chronic renal disease (CRD) patients with a normal range serum creatinine. Studies of this potential relationship have been limited by failure to assess true GFR, failure to assess proteinuria in a quantitative manner, or arbitrary restriction of the range of proteinuria examined. We examined the potential independent relationship between plasma tHcy levels and a wide range of quantitatively determined proteinuria (i.e., 0.000-8.340 g/day), among 109 CRD patients with a normal range serum creatinine (range; 0.8-1.5 mg/dl; median=1.2 mg/dl). Glomerular filtration rate (GFR) was directly assessed by iohexol clearance, and plasma status of folate, pyridoxal 5'-phosphate, and B12, along with serum albumin, were also determined. Linear modeling with ANCOVA revealed that proteinuria was not independently associated with tHcy levels (partial R=0.127; P=0.201), after adjustment for potential confounding by GFR (partial R=0.408; P<0.001), age, sex, plasma B-vitamin status, and serum albumin. Moreover, descending across quartiles (Q) [from Q4 to Q1] of GFR, ANCOVA-adjusted (i.e., for age, sex, and folate status) geometric mean tHcy levels (micromol/l) were significantly increased: tHcy Q4 GFR=9.6; tHcy Q3 GFR=10.5; tHcy Q2 GFR=11.9; tHcy Q4 GFR=14.5; P<0.001 for overall Q difference. We conclude that across a broad spectrum of quantitatively determined proteinuria, after adjustment for true GFR, in particular, there is no independent relationship between proteinuria and tHcy levels among CRD patients with a normal range serum creatinine.

Adult↗

Homocyst(e)ine metabolism in hemodialysis patients treated with vitamins B6, B12 and folate.

Hyperhomocyst(e)inemia is commonly accepted as an independent atherosclerotic risk factor. In most hemodialysis patients, serum homocyst(e)ine is markedly elevated and may contribute to premature atherosclerosis in these patients. Whereas the beneficial effect of folate supplementation on serum homocyst(e)ine has been extensively studied, there are less detailed studies on the effects of cobalamin and pyridoxal phosphate alone, or in combination with folate. We examined the effect of a four-week course of intravenous treatment with folate (1.1 mg), cobalamin (1.0 mg), and pyridoxal phosphate (5.0 mg), administered once (group 1), twice (group 2) or thrice (group 3) weekly in 33 hemodialysis patients divided in three groups of 11 patients. All patients were followed for a further four weeks after treatment was stopped. Serum homocyst(e)ine, cobalamin, folate and pyridoxal phosphate, as well as the metabolites of homocyst(e)ine, methylmalonate, 2-methylcitrate and cystathionine, were determined before, during and after treatment. Baseline serum homocyst(e)ine correlated significantly with serum folate (P=0.0149), cobalamin (P=0.0047) and pyridoxal phosphate (P=0.0408). Correlations independent from the other metabolites or vitamins were found for methylmalonate (P=0.003) and folate (P=0.029). All regimens increased serum cobalamin significantly (in group 1 from 444 +/- 215 to 17,303 +/- 11,989 pg/ml, P<0.01; in group 2 from 542 +/- 633 to 44,896 +/- 15,772 pg/ml, P<0.001; in group 3 from 548 +/- 394 to 77,961 +/- 31,546 pg/ml, P<0.001), but did not increase any of the other vitamin levels. Serum homocyst(e)ine was lowered significantly by 39.8% +/- 31.9% (P<0.05) with two and by 30.1% +/- 26.9% (P<0.05) with three vitamin dosages weekly, but not with one dosage weekly. Since methylmalonate is known to be a sensitive marker of cobalamin deficiency, the data support an important influence of cobalamin levels on baseline homocyst(e)ine levels. Increasing cobalamin levels and additional treatment with folate and pyridoxal phosphate 156 may decrease serum homocyst(e)ine in the same way as high doses of folate alone.

Female↗

Homocysteine, cysteine, and B vitamins as predictors of kidney disease progression.

BACKGROUND: Pathological similarities between atherosclerosis and glomerulosclerosis suggest that risk factors for the two processes may be similar. Elevated total homocysteine (tHcy) levels and low B vitamin levels are risk factors for atherosclerosis, but have not been evaluated sufficiently as risk factors for the progression of kidney disease. METHODS: Frozen samples from the Modification of Diet in Renal Disease Study were assayed for serum tHcy, cysteine, pyridoxal 5-phosphate (PLP), folate, and vitamin B12 levels in 804 participants. These factors were evaluated in both continuous and categorical analyses as risk factors for glomerular filtration rate (GFR) decline by using univariate and multivariable analyses. RESULTS: At baseline, mean tHcy levels in study A (GFR, 25 to 55 mL/min/1.73 m2) and study B (GFR, 13 to 24 mL/min/1.73 m2) were 16.9 micromol/L (median, 15.6 micromol/L) and 23.0 micromol/L (median, 20.5 micromol/L), respectively. Mean follow-up was 2.2 years. Mean GFR declines were -4.35 and -3.65 mL/min/y in studies A and B, respectively. There was no significant association between change in GFR with baseline level of tHcy in univariate (-0.26 mL/min/y per 1-SD unit increase in tHcy level; 95% confidence interval [CI], -0.67 to 0.15) or multivariable (-0.18 mL/min/y per 1-SD unit increase in tHcy level; 95% CI, -0.53 to 0.17) analysis in study A or univariate (0.07 mL/min/y per 1-SD unit increase in tHcy level; 95% CI, -0.36 to 0.51) or multivariable (0.24 mL/min/y per 1-SD unit increase in tHcy level; 95% CI, -0.16 to 0.64) analysis in study B. Similarly, higher cysteine levels and lower B vitamin levels were not associated with faster rates of GFR decline in multivariable analysis in either study. CONCLUSION: Higher tHcy or cysteine levels and lower folate, PLP, and vitamin B12 levels are not independent risk factors for progression of nondiabetic kidney disease.

Adolescent↗

Tissue distribution and prediction of total body folate of rats.

To clarify relationships between dietary folic acid intake, blood levels and body stores of folate, rats were fed an amino acid-based diet supplemented with 0, 0.125, 0.5, 1, 2 or 4 mg folic acid/kg diet for 25 d. Folate concentrations of carcass, liver, gastrointestinal (GI) tract, kidney, spleen, testes, heart and lung from rats fed the folate-free diet were 0.06 +/- 0.01, 0.73 +/- 0.08, 0.05 +/- 0.01, 0.39 +/- 0.01, 0.05 +/- 0.01, 0.17 +/- 0.01, 0.02 +/- 0.01 and 0.02 +/- 0.01 micrograms/g, respectively. Serum and erythrocyte concentrations and total body stores were 0.88 +/- 0.16 ng/mL, 0.30 +/- 0.01 micrograms/mL and 13.9 +/- 0.7 micrograms, respectively. Body folate distribution was carcass, 55.6 +/- 1.4%; liver, 26.0 +/- 1.9%; erythrocytes, 7.7 +/- 0.4%; kidney, 4.8 +/- 0.2%; GI tract, 3.0 +/- 0.2%; and testes, 2.5 +/- 0.2%. Carcass content dropped to 38% whereas liver content increased to 44% in rats fed the highest dietary level. Tissue concentrations were correlated with one another and with dietary folate levels. Under these experimental conditions total body folate could be predicted from serum folate, but the general applicability of this relationship requires further study.

Animals↗

Effects of short-term supplementation with ascorbate, folate, and vitamins B6 and B12 on inflammatory factors and estrogen levels in obese postmenopausal women.

Little is known about the effects of commonly used vitamins on serum inflammatory markers and the hormonal balance in obese postmenopausal women. We studied the effects of an 8-week open-label supplementation with vitamins C (500 mg), B6 (25 mg), B12 (1 mg), and folate (5 mg) on C-reactive protein, interleukin-6, and estradiol levels in 20 obese (body mass index > or = 30) postmenopausal women. Outcomes were assessed in a blinded fashion. Folate and vitamin B12 levels rose significantly, suggesting that the supplement was well absorbed and that participants adhered to the protocol. Weight, blood pressure, and serum lipids remained stable. C-reactive protein, interleukin-6, and leptin levels remained unchanged. Estradiol levels rose from a median of 22.0 pg/mL (IQR = 15.9-25.8) at baseline to a median of 27.8 pg/mL (IQR = 23.1-33.9) at follow-up (p = 0.003). Increments in serum estradiol caused by vitamin supplementation in postmenopausal women have not been previously described and probably merit further investigation.

Ascorbic Acid↗

Comparing biological measurements of vitamin C, folate, alpha-tocopherol and carotene with 24-hour dietary recall information in nonhispanic blacks and whites.

PURPOSE: To assess intake of several vitamins in preparation for a large cohort study investigating the effect of diet on risk of colon and prostate cancer. METHODS: The dietary intake of several vitamins were assessed using eight different 24-hour recalls and a 200-item food frequency questionnaire (FFQ) from each subject. Participants also attended a clinic where blood was drawn and body composition, weight, height, and blood pressure were measured. A total of 97 black and 96 nonhispanic white subjects participated. The levels of alpha-tocopherol, carotene, folate, and vitamin C in the blood were correlated with the dietary intakes as measured by both 24-hour recalls and FFQ. RESULTS: Correlations between blood levels and energy-adjusted dietary intake assessed by 24-hour recalls (with supplements) were as follows: carotene (adjusted for serum cholesterol): 0.47 and 0.55 in black and white subjects, respectively; alpha-tocopherol (adjusted for serum cholesterol): 0.61 (blacks) and 0.50 (whites); vitamin C: 0.22 (blacks) and 0.17 (whites); folate: 0.54 (blacks) and 0.55 (whites). Correlations between blood levels and FFQ indices were smaller in magnitude: 0.34 and 0.28 for carotene in black and white subjects, respectively, 0.37 and 0.56 for alpha-tocopherol (adjusted for serum cholesterol), 0.20 and 0.03 for vitamin C and 0.24 and 0.32 for folate. CONCLUSIONS: The correlations observed were generally of modest to moderate size and were similar to or larger than those reported by others. This is despite variations in absorption, metabolism, and excretion of the vitamins and suggests that both the 24-hour recalls and the FFQ contain valid information.

Black or African American↗

Oral contraceptives can cause falsely low vitamin B(12) levels.

Serum vitamin B(12) radioimmunoassays may give falsely low results in patients with folate deficiency, multiple myeloma, megadose of vitamin C and following radioisotope organ scan. We evaluated 10 consecutive healthy women on oral contraceptives (OC) who had falsely low vitamin B(12) levels, as reflected by normal urine methylmalonic acid and plasma homocysteine. After 1-month cessation of OCs, vitamin B(12) returned to the normal range in all women. Transcobalamin I (TCI) blood level was decreased in 60% of patients. OCs may cause temporary low vitamin B(12) blood levels of no clinical significance that can be associated with low TCI levels

Adult↗

Supplementation with mixed fruit and vegetable juice concentrates increased serum antioxidants and folate in healthy adults.

OBJECTIVE: Epidemiological studies have shown that low plasma levels of antioxidant micronutrients, which are commonly found in fruit and vegetables, are associated with increased risk for diseases such as heart disease, cancer, metabolic disorders and the like. The aim of this study was to monitor the dietary habits of a group of healthy, middle-aged, men and women and to assess the effect of supplementation with a natural phytonutrient preparation from fruits and vegetables, on plasma levels of various antioxidant micronutrients and oxidative stress assessed by measuring 8-oxodGuo (8-oxo-7,8-dihydro-2'-deoxyguanosine) in urine. METHODS: The study followed a double-blind randomized cross-over design involving 59 healthy men and women (40-60 years of age). The supplement or a placebo was given to two groups for a total period of 14 weeks (crossover week 7). Blood levels of beta-carotene, vitamin C, vitamin E, selenium and folate were measured at 0, 7 and 14 weeks. Fruit and vegetable consumption was monitored by means of a retrospective food frequency questionnaire at week 0, 7 and 14. Urinary 8-oxodGuo was also determined at these time points. RESULTS: Significant increases in blood nutrient levels after active supplementation were observed for beta-carotene, vitamin C, vitamin E, selenium and folate. Ranges measured, after supplementation, often fell into those associated with a reduced risk for disease. Our data suggests that, although generally health conscious, participants still fell short of the recommended five portions of fruit and vegetables per day. No significant group changes were noted for 8-oxodGuo concentration in urine. CONCLUSION: Supplementation with mixed fruit and vegetable juice concentrates effectively increased plasma levels of important antioxidant nutrients and folate.

8-Hydroxy-2'-Deoxyguanosine↗

Extensive laboratory assessment of nutritional status in fit, health-conscious, elderly people living in the Paris area. Research Group on Aging.

OBJECTIVE: Nutritional status-related biological indexes were measured in fit, health-conscious elderly subjects in order to establish reference values for people over 70 years. SUBJECTS: 103 men and 90 women aged 70-89 years living freely in the Paris area volunteered to participate. METHODS: Nutritional status was assessed by anthropometric and biochemical methods. RESULTS: Serum protein and amino acid status was similar to that of young adults, with only 5.2% of the elderly subjects showing transthyretin concentrations < 0.20 g/L, as well as decreased essential amino acid levels. Iron status, assessed in terms of serum and erythrocyte ferritin levels, total iron binding capacity and erythrocyte protoporphyrin tended to be satisfactory, but iron depletion was detected in 8.8% of the subjects. Serum ferritin levels were elevated in 19.7% of the subjects. Folate and vitamin B12 status was satisfactory, while hypovitaminosis D was observed in 48.2% of cases. CONCLUSION: Our findings suggest that, in aging uncomplicated by disease, nutritional status is similar to that in younger adults, although the range of values tended to be wider, with a higher risk of certain nutrient deficiencies.

Aged↗

Purification of folate binding factor in normal umbilical cord serum.

Human umbilical cord serum was found to contain both free folate and folate complexed to a high-molecular weight factor. The complexed folate was bound to a very high affinity binder and was present in concentrations equivalent to as much as 60 ng of 5-methyltetrahydrofolic acid per ml of serum. Acidification of the serum caused disassociation of the folate-binder complex. Released folates were separated from binder by Sephadex gel filtration, zonal centrifugation through sucrose gradients, or adsorption onto activated charcoal. The separated binding factor, either saturated or unsaturated with folate, had a molecular weight of about 40,000 on Sephadex G-200 chromatography. Binding of [3H]pteroylglutamic acid was rapid and, as in the original endogenous folate-binder complex, was essentially irreversible at neutral pH. The affinity and specificity of the binder were examined by competition experiments using [3H]pteroylglutamic acid and nonradioactive folate derivatives. Oxidized folates were bound in preference to reduced derivatives, but only three to four times more unlabeled 5-methyltetrahydrofolic acid than pteroylglutamic acid was required to produce an equal level of competition. The strong affinity for 5-methyltetrahydrofolic acid, the main serum folate, suggests that the binder could be part of the mechanism by which the fetus concentrates maternally supplied folate for its growth and development.

Binding Sites↗

Investigation of factors influencing folate status in women who have had a neural tube defect-affected infant.

OBJECTIVE: To investigate folate intake and blood levels of vitamins in women with and without a history of neural tube defect (NTD)-affected infant and to explore the relation between red cell and serum folate in those women. SUBJECTS: Twenty-nine women with a history of NTD-affected infant, who had taken periconceptional vitamin supplements one year or more prior to the study and twenty-nine controls with no such history. MAIN OUTCOME MEASURES: Dietary intake of folate was assessed by questionnaire and blood levels of folate, vitamin B12 and vitamin C were measured. RESULTS: Neither dietary intake of folate nor any of the blood vitamins measured were lower in the women with a history of NTD infant. The majority of women who had received periconceptional vitamin supplementation subsequently had adequate folate intakes and red blood cell folates greater than 160 micrograms/l. Smokers had lower plasma Vitamin C levels than nonsmokers. Multiple regression analysis suggested that the relation between red blood cell (RBC) and serum folate may differ between women with and without a history of NTD. CONCLUSIONS: Women with a history of NTD-affected infants may have different folate metabolism from those who have not.

Adult↗

The use of biomarkers in multicentric studies with particular consideration of iodine, sodium, iron, folate and vitamin D.

OBJECTIVE: To determine which biomarkers should be used in a European monitoring system to assess intake of iodine, sodium, iron, folate and vitamin D. These nutrients are relevant for human health but difficult to assess in the diet in a comparable way among countries. RESULTS: Iodine and sodium are mainly excreted in urine. Mean population intakes of iodine and sodium can be estimated by casual urine samples. However, habitual individual intake assessments require multiple 24 h urine collections. Serum ferritin is a sensitive test of iron status (mobilizable storage iron) and continues to be the leading single determination for individual iron status, but with serum transferrin receptor as a promising alternative because it is more specific than ferritin. Erythrocyte folate and homocysteine in blood are markers of folate status which are able to discriminate between normal status and levels of insufficiency in an individual. Serum 25-hydroxyvitamin D is a good marker for vitamin D deficiency in the individual, and can distinguish between overt deficiency and marginal deficiency, especially if combined with serum parathyroid hormone. CONCLUSIONS: Specific and sensitive markers for the dietary intake of iodine, sodium, iron, folate and vitamin D are available, which have the advantage of being more accurate than intake estimations derived from dietary surveys. However, the use of biomarkers also has disadvantages such as extra burden in terms of logistics, budget and ethical approval, and can moreover reduce the participation rate. More basic data for the validity of cross-country comparisons are needed and ideas in terms of study design and sampling frame.

Biomarkers↗

Assessment of nutritional folate status and selected vitamin status of women of childbearing age.

OBJECTIVE: The aim of the study was to investigate folate and other selected vitamin status (ascorbic acid, tocopherol, retinol, vitamin B12), haematological indices and total homocysteine concentration of serum in women of childbearing age. DESIGN: A cross-sectional study. SETTING: Warsaw. SUBJECTS: Healthy women aged 18-30 y (n=78) not pregnant presently or previously, and not taking drugs. RESULTS: Haemoglobin and haematocrit values according to WHO criteria for 18 to 30-y-old women were normal. The optimal levels of serum tocopherol, >1.29 mg/dl (>29.9 micromol/l) to preventing civilization diseases, were found in 5.5% and serum retinol >71.6 mcg/dl (>2.5 micromol/l) in 6.4% of all studied persons. The analysis of serum folate concentration showed high-risk deficiency,<3 ng/ml (<6.8 nmol/l), in 6.4%, moderate and low risk together (7.0-14.9 nmol/l) in 61.6% and optimal folate levels (>14.9 nmol/l) in 32.0% of the studied group. Folate body stores were insufficient in almost all women. There was no high or moderate deficiency risk of vitamin B12 or ascorbic acid. None of the women under study had serum total homocysteine (tHcy) concentration >15 micromol/l, indicating hyperhomocysteinaemia. Serum total homocysteine concentrations in the range of 5-15 micromol/l were found in 71.8%, and serum tHcy >10 micromol/l in 7.7% of the studied group of women. SPONSORSHIP: The study was sponsored by the Polish Committee for Scientific Research.

Adolescent↗

Alcohol, one-carbon metabolism, and colorectal cancer: recent insights from molecular studies.

A growing body of evidence implicates alcohol intake as a risk factor for colorectal cancer. Until recently, most of the data were based on epidemiologic data that examined alcohol intake in relation to risk of colorectal neoplasia, but the evidence has now been broadened by recent molecular studies on mechanisms. In particular, a number of observations strongly support a role for alcohol acting through disruptions in one-carbon metabolism. Excessive alcohol intake is a fairly consistent risk factor for colorectal neoplasia in most studies, and studies show much higher risks of colorectal neoplasia in those with high alcohol and low folate than with high alcohol or low folate individually. Interactions between high alcohol-low folate and the MTHFR677TT genotype with risk of colorectal neoplasia suggest that alcohol is acting through its effects on one-carbon metabolism because the combination of high alcohol-low folate and the MTHFR677TT genotype are related to markedly elevated serum levels of homocysteine and to DNA hypomethylation. In addition, in Japanese studies, consumers of alcohol possessing the ALDH2*2 allele, who have very elevated levels of acetaldehyde, are at high risk for colorectal cancer. The relatively high prevalence of the ALDH2*2 allele may thus account for the stronger association between alcohol and colorectal neoplasia that is frequently observed in studies in Japanese populations. Further research integrating studies with more detailed data on alcohol intake levels and patterns, folate and other related nutrient status, and relevant genotypes may help clarify unresolved questions regarding the health consequences of moderate to high alcohol drinking.

Alcohol Drinking↗

Zinc status is not adversely affected by folic acid supplementation and zinc intake does not impair folate utilization in human subjects.

Changes in zinc status in response to folic acid supplementation and the effect of zinc intake on folate utilization were evaluated in 12 men (20-34 y old) consuming a diet containing 3.5 or 14.5 mg zinc/d for two 25-d intervals. Deuterium-labeled folic acid (800 micrograms/d) or a placebo was administered orally during each phase. No differences in plasma zinc, erythrocyte zinc, urinary zinc, erythrocyte metallothionein or serum alkaline phosphatase, due to supplemental folic acid, were detected at either level of zinc intake. Differences in the response to folic acid supplementation, due to the level of zinc intake, were not detected for serum, erythrocyte or urinary (labeled and unlabeled) folate. Within the constraints of this short-term folic acid supplementation study, adverse effects on zinc status were not observed and our data suggest that folic acid utilization was not influenced by level of zinc intake.

Administration, Oral↗

Changes in folate status as determined by reduction in total plasma homocysteine levels during leucovorin modulation of 5-fluorouracil therapy in cancer patients.

We measured plasma total homocysteine (tHcy) in 14 patients (13 patients with colorectal cancer and 1 patient with breast cancer) during their first treatment with 5-fluorouracil (5-FU) plus leucovorin [LV (5-FULV)]. Eight of these patients were investigated a second time after 3-10 cycles (median, 4 cycles) with 5-FULV. Each cycle consisted of two administrations of 5-FU (500 mg/m2) and LV (60 mg/m2) given 24 h apart. The first administration of 5-FULV on day 1 of the first cycle induced a rapid reduction of the tHcy level from 12.5 micromol/liter (10.4-15.1 micromol/liter; geometric mean with 95% confidence interval of the mean) to 9.1 micromol/liter (7.5-11.1 micromol/liter) in 24 h. tHcy remained stable at this level after the second administration of 5-FULV. In addition, the 5-FULV regimen caused a concurrent 4-fold increase in both serum and erythrocyte folate. The fifth cycle with 5-FULV had only marginal effects on the tHcy level. 5-FU without LV modulation had no effect on the plasma tHcy or folate status in eight breast cancer patients. Our data establish the reduction of tHcy as a responsive indicator of LV pharmacodynamics.

Aged↗

Homocysteine and folate levels as risk factors for recurrent early pregnancy loss.

OBJECTIVE: To estimate the relative risk of recurrent early pregnancy loss for different total plasma homocysteine and serum folate concentrations. METHODS: In a case-control study, we measured homocysteine (fasting and afterload), folate (serum and red cells), pyridoxal 5'-phosphate, and cobalamin concentrations in 123 women who had at least two consecutive spontaneous early pregnancy losses each and compared concentrations with those of 104 healthy controls. RESULTS: Women with recurrent early pregnancy losses had significantly lower serum folate concentrations than controls, whereas the other measurements were similar to those of controls. Elevated homocysteine, fasting greater than 18.3 micromol/L and afterload greater than 61.5 micromol/L, was a risk factor for recurrent early pregnancy loss, with odds ratios (ORs) and 95% confidence intervals (95% CIs) of 3.6 (1.2, 12.7) and 2.7 (0.9, 8.8) in the group with recurrent miscarriages: 6.4 (1.9, 24.3) and 4.3 (1. 2, 17.3) in primary aborters, and 4.2 (1.3, 15.4) and 3.4 (1.0, 12. 8) in those with three or more miscarriages. The ORs (95% CIs) in the same study populations for serum folate concentrations less than 8.4 nmol/L were 2.1 (0.9, 4.8), 2.7 (1.0, 7.8), and 3.2 (1.3, 8.1), respectively. A significant dose-response relationship between serum folate concentrations and risk of recurrent early pregnancy loss suggested a protective effect by high serum folate concentrations. CONCLUSION: Elevated homocysteine and reduced serum folate concentrations were risk factors for recurrent spontaneous early pregnancy losses. Folic acid supplementation might be beneficial in women with histories of early pregnancy loss.

Abortion, Habitual↗