Objective:To investigate whether vitamin B(6) supplementation has a beneficial effect on immune responses in critically ill patients.Design:A single-blind intervention study.Setting:The study was performed at the Taichung Veterans General Hospital, the central part of Taiwan.Subjects:Fifty-one subjects who stayed over 14 days in the intensive care unit completed the study. Subjects were not treated with any vitamin supplement before the intervention.Interventions:Patients were randomly assigned to one of three groups, control (n=20), a daily injection of 50 mg vitamin B-6 (B(6)-50, n=15), or 100 mg vitamin B-6 (B(6)-100, n=16) for 14 days.Main outcome measures:Plasma pyridoxal 5'-phosphate (PLP), pyridoxal (PL), 4-pyridoxic acid (4-PA), erythrocyte alanine (EALT-AC) and aspartate (EAST-AC) aminotransaminase activity coefficient, and urinary 4-PA were measured. The levels of serum albumin, hemoglobin, hematocrit, high-sensitivity C-reactive protein (hs-CRP) and immune responses (white blood cell, neutrophils, total lymphocytes count (TLC), T-(CD3) and B-(CD19) lymphocytes, T-helper (CD4) and suppressor (CD8) cells) were determined.Results:Plasma PLP, PL, 4-PA and urinary 4-PA concentrations significantly increased in two treated groups. T-lymphocyte and T-helper cell numbers and the percentage of T-suppressor cell significantly increased on day 14 in the B(6)-50 group. Total lymphocyte count, T-helper and T-suppressor cell numbers, the percentage of T-lymphocyte cells and T-suppressors significantly increased in the B(6)-100 group at the 14th day. There were no significant changes with respect to immune responses in the control group over 14 days.Conclusions:A large dose of vitamin B(6) supplementation (50 or 100 mg/day) could compensate for the lack of responsiveness of plasma PLP to vitamin B(6) intake, and further increase immune response of critically ill patients.Sponsorship:This study was supported by the National Science Council, Taiwan, Republic of China (NSC-92-2320-B-040-026).European Journal of Clinical Nutrition advance online publication, 3 May 2006; doi:10.1038/sj.ejcn.1602439.

OBJECTIVE:: To investigate whether vitamin B(6) supplementation had a beneficial effect on lowering fasting plasma homocysteine concentrations in coronary artery disease (CAD) patients. DESIGN:: A single-blind intervention study. SETTING:: The study was performed at the Taichung Veterans General Hospital, the central part of Taiwan. SUBJECTS:: A total of 50 subjects were identified by cardiac catheterization to have at least 70% stenosis of one major coronary artery. In all, 42 patients successfully completed this study. INTERVENTIONS:: Patients were randomly assigned to one of five groups and treated with a daily dose of placebo (n=8), 5 mg vitamin B(6)(n=8), 10 mg vitamin B(6)(n=8), 50 mg vitamin B(6)(n=9), or 5 mg folic acid combined with 0.25 mg vitamin B(12)(n=9) for 12 weeks. MAIN OUTCOME MEASURES:: Nutrient intakes were recorded by using 24-h diet recalls when patients returned to the cardiology clinic before the intervention (week 0) and at week 12. Vitamin B(6) status was assessed by direct measures (plasma pyridoxal 5'-phosphate) and indirect measures (erythrocyte alanine and aspartate aminotransaminase activity coefficient). Fasting plasma homocysteine, serum folic acid, and vitamin B(12) were measured. RESULTS:: Fasting plasma homocysteine concentration did not respond to high or low doses of vitamin B(6) when compared with a placebo treatment after 12 weeks of supplementation. The mean fasting plasma homocysteine concentration, however, decreased significantly after 12 weeks of folic acid combined with vitamin B(12) supplementation (P=0.047). Further, within group, mean fasting plasma homocysteine concentration was nonsignificantly increased by 25.5, 16.2, and 18.3% in placebo, 10 mg/day and 50 mg/day vitamin B(6) supplemented groups, respectively; whereas folic acid combined with vitamin B(12) supplementation significantly reduced fasting plasma homocysteine concentration by 32%(P<0.001). CONCLUSIONS:: Our results indicate that vitamin B(6) supplementation alone is less effective than folic acid combined with vitamin B(12) in lowering plasma homocysteine concentrations in CAD patients. SPONSORSHIP:: This study was supported by the National Science Council, Taiwan, Republic of China (NSC-91-2320-B-040-023).European Journal of Clinical Nutrition (2004) 58, 481-487. doi:10.1038/sj.ejcn.1601834

Accurate determination of energy expenditure is essential in patients receiving nutritional support to meet metabolic needs. The purpose of this study was to assess and compare the energy expenditure as measured by indirect calorimetry (MEE) and estimated by 5 equations in the mechanically ventilated critically ill patients. Forty-six patients were divided into either enteral nutrition (EN)(n=l2), total parenteral nutrition (TPN)(n=16) or combined (EN plus TPN)(n=l8) groups. Patients' energy expenditure was measured by indirect calorimetry on two occasions. Anthropometric and biochemical measurements, energy expenditure and medical status (APACHE II score) were also assessed in the intensive care unit (ICU) of Taichung Veteran General hospital. No significant difference was found in the MEE among the 3 groups. The type of nutritional support did not affect MEE. Energy expenditure calculated by using Harris- Benedict, Kleiber and Liu equations times the estimated stress factor did not significantly different than the values of MEE in all groups. There were significant correlations (P<0.01) between MEE and patients' sex (r=-0.499), age (r=-0.402), height (r=0.533), knee height (r=0.431), current body weight (r=0.379), usual body weight (r=0.407), ideal body weight (r=0.466) and urinary urea nitrogen (r=0.383) in the pooled group. Results demonstrated that energy expenditure could be estimated in most critically ill patients by using Harris-Benedict, Kleiber and Liu equations if the estimated stress factor is in the reasonable value.

OBJECTIVE: To assess vitamin B6 intake and status of critically ill patients. The relationship between vitamin B6 status indicators and the severity of illness and outcome in these patients was also examined. DESIGN: Prospective clinical study. SETTING: The study was performed at the Taichung Veteran General Hospital, in the central part of Taiwan. SUBJECTS: Ninety-four patients in the intensive care unit (ICU) entered the study and 46 patients successfully completed this study. INTERVENTIONS: No intervention. MAIN OUTCOME MEASURES: Vitamin B6 intake was recorded for 14 days. Vitamin B6 status was assessed by direct measures (plasma pyridoxal 5'-phosphate (PLP), pyridoxal (PL), and urinary 4-pyridoxic acid (4-PA)) and indirect measures (erythrocyte alanine (EALT-AC) and aspartate (EAST-AC) aminotransaminase activity coefficient). The severity of illness (APACHE II score), the length of ventilation dependency, and the length of ICU and hospital stay were recorded. RESULTS: Patients had an adequate mean vitamin B6 intake (16.26+/-19.39 mg) during the 14 day study. Mean vitamin B6 intake was significantly higher on day 14 than on day 1 (P<0.001). However, plasma PLP and PL concentrations significantly decreased at the 14th day after admission (P<0.05). Erythrocyte alanine aminotransaminase activity coefficient and EAST-AC did not change significantly. Urinary 4-PA significantly increased at the 14th day (P<0.001). No significant relationships were found between APACHE II scores and clinical outcomes (the length of ICU and hospital stay, the length of ventilation dependency) of patients, vitamin B6 intake or status indicators. CONCLUSIONS: Critically ill patients received nutritional support in the ICU, and had sufficient mean vitamin B6 intake and adequate vitamin B6 status. Therefore, the severity of illness and the results should not be affected by vitamin B6 status. However, we have noted that plasma PLP and PL concentrations significantly decreased while vitamin B6 intake significantly increased on day 14. Critical clinical conditions and complex metabolism in the critically ill may account for the reduction of plasma PLP and PL. Since vitamin B6 deficiency causes profound effects on immune system function, dietary or supplemented vitamin B6 intake is suggested for hospitalized patients.

SETTING: A cohort of 78 adolescents was selected for evaluation with culture or histologically proven pulmonary tuberculosis (PTB) from a tertiary paediatric facility in northern Taiwan. OBJECTIVE: To assess the validity of clinical features and radiographic findings for predicting positive smears of acid-fast bacilli (AFB) in adolescents with PTB. DESIGN: A retrospective descriptive study of adolescents with a confirmed diagnosis of PTB. Clinical symptoms and chest radiographs were assessed. Univariate analysis identified risk factors suggestive of a positive AFB smear, and the adjusted odds ratio (aOR) for these features was calculated using logistic regression. RESULTS: Patients who were AFB smear-positive and those who were smear-negative differed significantly on univariate analysis (P < 0.05) with respect to chronic cough, haemoptysis, multilobar or superior segment of lower lobe involvement, cavitations or presence of pleural effusions. Logistic regression analysis revealed that risk factors of positive smear in adolescents with PTB were chronic cough >4 weeks (aOR 13.8, 95%CI 2.3-83.1), lower lobe involvement (aOR 12.6, 95%CI 1.2-134.8) and pulmonary cavitations (aOR 7.7, 95%CI 1.0-57.7). CONCLUSIONS: For adolescents with PTB, those suffering from chronic cough for >4 weeks, with involvement of the superior segment of the lower lobe or with cavitary lesions, have a greater likelihood of transmitting tuberculosis due to smear positivity.

XRCC1 plays a central role in mammalian DNA repair processes. Two polymorphisms of XRCC1, rs1799782 (Arg > Trp at codon 194) and rs25487 (Arg > Gln at codon 399), are common in the Han Chinese population. Our objective was to analyze the relationship between these two functional single-nucleotide polymorphisms (SNPs) and systemic lupus erythematosus (SLE) in the Taiwanese Han Chinese population. Genotyping was performed by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) on 172 SLE patients and 160 normal controls. Our data indicate that the frequency of A/G at codon 399 differed between patients and controls (p = 0.01; odds ratio: 1.80; 95% confidence interval: 1.17-2.75), but the allelic frequency analysis did not reveal significant differences. For the SNP at codon 194, there were no differences in either allelic or genotype frequencies between SLE patients and normal subjects. Clinical association studies of SLE symptoms revealed the involvement of the A/G polymorphism at codon 399 in SLE pathogenesis. Our results indicate that a functional SNP at codon 399 of XRCC1 is associated with the development of SLE.

Clin Microbiol InfectAbstract Levofloxacin susceptibility testing was carried out for a total of 2539 Streptococcus pneumoniae isolates obtained from January 2001 to February 2008 at the National Taiwan University Hospital (NTUH) and a further 228 pneumococcal isolates obtained from January 2004 to December 2006 at three other hospitals in different geographical areas in Taiwan. Levofloxacin non-susceptible S. pneumoniae isolates were subsequently analysed for serotype and molecular epidemiology. Rates of levofloxacin non-susceptibility of S. pneumoniae increased significantly from 1.2% in 2001 to 4.2% in 2007 at NTUH. A total of 30 isolates of levofloxacin non-susceptible S. pneumoniae isolates (MIC >/= 4 mg/L) were available for evaluation of serotype, antimicrobial susceptibility, nucleotide sequence of the quinolone resistance-determining regions of parC, gyrA, parE and gyrB, reserpine effect on quinolone susceptibility and multilocus sequence type. Among these isolates, seven (23.3%) were from children, and two (6.7%; one from a 3- and one from a 93-year-old patient) were from blood. One levofloxacin-resistant isolate (MIC = 8 mg/L) was recovered from a previously healthy child with bacteraemic necrotizing pneumonia complicated by empyema and a haemolytic-uraemic syndrome. All isolates except two had Ser79 and/or Asp83 changes in ParC, and/or Ser81 or Glu85 changes in GyrA. An efflux phenotype concerning levofloxacin was detected in only one (3.3%) isolate. A novel clone (ST3642), genetically related to Spain(9V)-3 and belonging to serotype 11A, was identified. Dissemination of clonal complexes related to Spain(23F)-1, Taiwan(19F)-14, Spain(9V)-3 and Taiwan(23F)-15 has contributed to levofloxacin non-susceptibility among these S. pneumoniae isolates from Taiwan.

Intellectual disability (ID) is a common developmental disability observed in one to three percent of the human population. A possible role for the Angiotensin II type 2 receptor (AGTR2) in brain function, affecting learning, memory, and behavior, has been suggested in humans and rodents. Mice lacking the Agtr2 gene (Agtr2(-/y)) showed significant impairment in their spatial memory and exhibited abnormal dendritic spine morphology. To identify Agtr2 influenced genes and pathways, we performed whole genome microarray analysis on RNA isolated from brains of Agtr2(-/y) and control male mice at embryonic day 15 (E15) and postnatal day one (P1). The gene expression profiles of the Agtr2(-/y) brain samples were significantly different when compared to profiles of the age-matched control brains. We identified 62 differently expressed genes (p</=0.005) at E15 and in P1 brains of the Agtr2(-/y) mice. We verified the differential expression of several of these genes in brain samples using quantitative RT-PCR. Differentially expressed genes encode molecules involved in multiple cellular processes including microtubule functions associated with dendritic spine morphology. This study provides insight into Agtr2 influenced candidate genes and suggests that expression dysregulation of these genes may modulate Agtr2 actions in the brain that influences learning and memory.

Clin Microbiol InfectAbstract Accurate molecular surveillance is important in monitoring the dynamics of Streptococcus pneumoniae. A prospective study was conducted to collect invasive isolates of S. pneumoniae from children for genetic analysis from January 2004 to December 2006 in Taiwan. PCRs were performed to detect the zmpC and zmpD genes, both encoding a metalloprotease virulence factor in pneumococci, among these invasive isolates. During the study period, 68 invasive isolates of S. pneumoniae were obtained for analysis. Serotype 14 was the most common type isolated from children with invasive disease and was significantly associated with pneumonia (OR 3.1; 95% CI] 1.1-8.8; p 0.035). Serotype 23F was significantly associated with bacteraemia (OR 7.5; 95% CI 1.8-31.3; p 0.006). The seven-valent conjugate vaccine covered 83.8% of invasive isolates, but non-vaccine serotypes were more frequently isolated from patients with underlying diseases than from patients without underlying diseases (p 0.007 by Fisher's exact test). Clonal complexes related to international clones Spain23F ST81, Spain6B ST95, England14 ST9, Taiwan19F ST236, Taiwan23F ST242 and Colombia23F ST338 accounted for 52.9% of invasive isolates. Dissemination of the penicillin-resistant clones ST876, ST46, ST76 and ST2889, which were first identified in Taiwan, was also found; 1.5% of these invasive isolates carried the zmpC gene, and 47.1% of these invasive isolates carried the zmpD gene. In conclusion, the spread of certain international clones and some domestic antibiotic-resistant clones resulted in invasive diseases among Taiwanese children.

BACKGROUND/AIMS: 118 elderly participants (65-90 years) were assessed for any relationship between folate, related genes and hypertension. METHODS: Six B-vitamin-related SNPs were genotyped in 80 normotensive and 38 hypertensive subjects. RESULTS: Of six polymorphisms (677C>T-MTHFR, 1298A>C-MTHFR, 80G>A-RFC, 2756A>G-MS, 66A>G- MSR, 19bpDHFR and 1561C>T-GCPII), only 677C>T-MTHFR was a significant risk for hypertension: OR 1.89; 95% CI 1.07-3.32 (chi2 p = 0.038). Additionally, hypertensive subjects had a significantly lower intake of dietary folate than normotensive individuals (p = 0.0221), although this did not markedly alter blood metabolite levels. Several significant linear associations between dietary folate and related blood metabolites were found in normotensive subjects (p < 0.001 for Hcy, red cell and serum folate) and were as predicted on an a priori basis -- generally weaker associations existed in hypertensive subjects (p < 0.05 for serum folate). This was true for data examined collectively or by genotype. Multiple-regression analysis for diastolic or systolic blood pressure showed significant interaction for gender and folate intake (p = 0.014 and 0.019, respectively). In both cases this interaction occurred only in females, with higher folate intake associated with decreased blood pressure. Regressing diastolic blood pressure and 677C>T-MTHFR genotype showed significance (males; p = 0.032) and borderline significance (all subjects). CONCLUSION: Dietary folate and 677C>T-MTHFR genotype may modify blood pressure.

OBJECTIVE: The purpose of this study was to investigate the association of genetic polymorphism of cyclooxygenase-2 and prostacyclin synthase with myocardial infarction (MI) in Uigur population in Xinjiang. METHODS: 178 patients with MI and 175 healthy control subjects were detected on the genetic polymorphism of cyclooxygenase-2 and prostacyclin synthase by polymerase chain reaction-based restriction fragment length polymorphism. Other serum 6-keto-PGF1alpha concentration and biochemical indicators were detected in all the subjects. RESULTS:(1) The genotype distributions of the control group and MI group were in the Hardy-Weinberg equilibrium. The frequencies of CC, CA and AA genotype of prostacyclin synthase were 75.84%, 17.42% and 6.74% in MI group while they were 64.57%, 28.29% and 9.14% in controls respectively. There was significant difference in frequencies of CC genotype and C allele as well as CA and AA genotypes between controls and MI cases.(2) The frequencies of -765GG,-765GC and -765CC genotype of cyclooxygenase-2 were 78.65%, 19.66% and 1.69% in MI group while they were 55.43%, 34.86% and 9.71% in controls respectively. There was significant difference in frequencies of three genotypes and alleles between the two groups (P < 0.05 or P < 0.01).(3) In combined genotype analysis, the genotype of PGIS CC + COX-2 -765GG was significantly higher in patients with MI than in control subjects (P < 0.05). The odds ratio estimated through combined analysis of the PGIS CC and COX-2 -765GG genotypes (OR = 3.87) markedly increased when compared with that estimated separately from the PGIS CC (OR = 1.72) or COX-2 -765GG (OR = 2.94) genotype.(4) There was a significant difference in serum 6-keto-PGF1alpha level between MI group and control group (P < 0.05), but there were no differences found in every genotype of PGIS and COX-2 gene (P > 0.05). In the cases with both COX-2 -765GG and PGIS CC genotypes, the serum 6-keto-PGF1alpha levels was lower than that of others (P < 0.05). CONCLUSION: The CC genotype and C allele of prostacyclin synthase,-765GG genotype and G allele of COX-2 might serve as risk factors of MI of Uigur population in Xinjiang. Populations with both COX-2 -765GG and PGIS CC genotypes were more at risk with MI than others which might be resulted from the decreased serum 6-keto-PGF1alpha concentration. The -765CC genotype and C allele of COX-2 gene might have protective functions on MI among Uigur population in Xinjiang.

OBJECTIVE: To determine whether genetic variations in methylenetetrahydrofolate reductase (MTHFR) are associated with the risk of laryngeal squamous cell carcinoma (LSCC) in a Chinese population. METHODS: Two hundred and seven cases with LSCC and 400 matched health controls were genotyped for the MTHFR 677C > T and 1298A > C polymorphisms by PCR-restriction fragment length polymorphism (PCR-RFLP) methods. The relation between these genotypes and risk of LSCC and gene-environment interaction were analyed. The adjusted odds ratios (OR) and 95% confidence intervals (CI) were calculated by using unconditional Logistic regression model. RESULTS: The individuals with 677CT and 677TT genotype had a 1.66-fold (95% CI 1.08-2.52) and 3.35-fold (95% CI 2.07-5.54) increased risk of developing LSCC comared with those who had 677CC genotype. The individuals with MTHFR 1298A > C genotype was not significantly different between the two groups. Furthermore, cigarette smoking was also found to interact with MTHFR 677C > T polymorphism in increasing the risk to LCSS further demonstrating the role of gene-environment interaction in development of LSCC. CONCLUSIONS: These findings suggested that the MTHFR 677C > T polymorphism may contribute to the risk of developing LSCC among Chinese population.

The 677C > T polymorphism of the MTHFR gene, resulting in hyperhomocysteinemia, has been shown to be implicated in the aetiology of schizophrenia. Previous studies showed that A1298 C polymorphism seems not to be related to schizophrenia. AIM OF THE STUDY: To analyze two genetic polymorphisms of the MTHFR gene, 677C > T and A1298 C in 44 patients with schizophrenia and evaluate its relationship with the risk of schizophrenia and with some clinical aspects. MATERIAL AND METHOD: We determined the presence of the 677C > T and A1298 C mutations of the MTHFR gene in 44 inpatients with schizophrenia and in 35 normal controls. The patients were assessed by psychiatric examination and scalar evaluation. RESULTS: 28 (66,7%) of the patient group had the T allele of the 677C > T genetic polymorphism, compared to 11 (34,3%) subjects of the control group. The intensity of the positive, negative and general symptoms was slightly higher in the patients presenting the T allele. The A1298C missense mutation was more frequent between control subjects (57,5%) compared to the patient group (39%). The intensity of the positive symptoms was slightly increased in the patients with the missense mutation in the position 1298, but the intensity of the negative and general symptoms did not differ. CONCLUSIONS: Our study confirms the role of the 677C > T genetic polymorphism in the susceptibility for schizophrenia. The relationship between A1298C genetic polymorphism and schizophrenia was not demonstrated in our study.

Fluoropyrimidine such as 5-fluorouracil(5-FU)exerts its antitumor activities via anabolism by several enzymes. Genetic polymorphisms of these enzymes related to sensitivity and toxicity of fluoropyrimidines are reviewed. Expression of thymidylate synthase(TS), a target enzyme of 5-FU, is regulated by variable number of a 28 bp tandem repeat in the enhancer region. The double tandem repeat is associated with low TS expression, and consequently, patients with double tandem repeat demonstrated higher sensitivity to 5-FU than those with triple tandem repeat. Single nucleotide polymorphism within the second tandem repeat and loss of heterozygosity are also reported to be related to fluoropyrimidine sensitivity. In addition, patients having a 6 bp deletion in 3'-UTR region showed remarkably high antitumor activity by 5-FU based chemotherapy. Genetic variations in 5-FU catabolic enzymes can also have a profound effect on 5-FU toxicity. So far 39 mutations/ polymorphisms have been identified in dihydropyrimidine dehydrogenase(DPD)gene, a major catabolic enzyme of 5- FU. Among them, IVS14+1G>A is reported to be highly associated with severe toxicity caused by chemotherapy with fluoropyrimidine. A polymorphism that may influence the efficacy of 5-FU by influencing folate pools is that of the methylenetetrahydrofolate reductase(MTHFR)gene. C677T mutation was associated with a higher response rate on 5-FU/folinic acid chemotherapy. Prospective clinical trials to confirm the predictability of genetic polymorphism for sensitivity and toxicity of 5-FU should be performed.

BACKGROUND: Hyperhomocysteinemia is known as an independent-risk factor for coronary-artery disease (CAD). However, the effect of homocystein metabolic enzymes polymorphisms on CAD is still controversed. We investigated the relation between homocystein metabolic key enzymes polymorphisms, homocystenemia and coronary stenosis in a Tunisian population. METHODS: Samples were collected from 251 CAD patients documented by angiography. Genotyping were performed for C677T methylene-tetrahydrofolate reductase (MTHFR), A2756G methionine-synthase (MS) and 844ins 68 cystathionine-beta-synthase (CBS). We measured fasting plasma tHcy, folate and vitamin B12. RESULTS: There was significant increase in homocysteinemia for homozygous genotypes of C677T MTHFR (p<0.001) and A2756G MS (p=0.01), but not for 844ins68 CBS (p=0.105). Potential confounders adjusted odds-ratios for significant coronary stenosis, associated with MTHFR TT, MS GG and CBS insertion, were respectively 1.78 (p=0.041); 2.33 (p=0.036) and 0.87 (p=0.823). The effect of mutated MTHFR genotype was more pronounced on homocysteinemia (21.4+/-9.1 micromol/L; p<0.001) and coronary stenosis (OR=2.73; p=0.033) at low folatemia (< or =6.1 ng/mL). CONCLUSION: MTHFR TT and MS GG genotypes increase tHcy concentration and coronary stenosis risk, especially with low folatemia.

BACKGROUND AND OBJECTIVE: Studies on different populations have shown a great variability of the frequencies of different polymorphisms in genes acting in the folate cycle. The present study was aimed to analyze the frequency in the Spanish population of each genotype combination of four polymorphisms, one of them -1561C-T of the glutamate carboxypeptidase II (GCPII) gene- being the first time that is studied in Spain. The study included a meta-analysis of the published data. SUBJECTS AND METHOD: Using the Spanish Collaborative Study of Congenital Malformations (ECEMC) Network, blood samples of 190 mother-child couples with newborns without any congenital defect, were obtained from 15 Spanish autonomous regions. The study polymorphisms were the 677C-T and 1298A-C polymorphisms of the methylenetetrahydrofolate reductase (MTHFR), the 66A-G of the methionine synthase reductase (MTRR), and the 1561C-T polymorphism of the GCPII gene. To estimate the range for the population frequencies, 99% confidence intervals were calculated. RESULTS: The frequencies observed in our country were significantly different from others, being similar to those obtained in countries of the Mediterranean European area. The 1561C-T polymorphism of the GCPII gene has a frequency in Spain of 5.11%, which is also similar to the values observed in France (5%) and in Italy (6%). On the other hand, the frequency of the genotypes CTCC, TTAC is quite few, while the genotype TTCC was not observed in any mother or infants. A meta-analysis was performed for a big sample (23,612 individuals) and the results showed that with a 99% of probability the values for the genotype combinations CTCC, TTAC, and TTCC were within 0.10-0.24; 0.20-0.36; and 0.003-0.05, respectively. CONCLUSIONS: Our results are important to further analyze the relationship with some health problems and individual susceptibilities. Indeed, considering the published observations of the structure and function of the MTHFR enzyme, it is understandable that those genotype combinations that are quite little frequent, may be related to the embryo-fetal viability, and to the life style of each population.

The aim of this work was to determine vitamin contents in spinach produced by different cultivation type (organic and traditional), harvesting period and after cooking in water. The determination was carried out by High Performance Liquid Chromatography. There was no significant difference in folate contents between spinach cultivated by traditional and organic method and there was also no significant difference between the values obtained at different periods of the year. Folate levels determined as 5-methylTHF and 5-formylTHF varied from 226 to 527 microg/100 g and 4.6 to 10 microg/100 g, respectively. Cooking in water resulted in approximately 74% of losses of 5-methylTHF and 56% of 5-formylTHF. The mainly losses occurred by leaching.

The objective of this study was to evaluate if there is any difference in the proportion of methylenetetrahydrofolate reductase (MTHFR) C677T polymorphisms and the homocysteine levels in a group of women with recurrent pregnancy loss (RPL) and a control group. Ninety-three patients with diagnosis of three or more gestational losses and 206 healthy women with two or more children, were included. After acceptance of informed consent, samples of peripheral blood were taken to determine the genetic polymorphisms of MTHFR C677T and the plasmatic levels of homocysteine. The carriers of the homozygous mutation TT of MTHFR 677T polymorphism were 12.9%(12 of 93) in the group of patients and 14.6%(30 of 206) in the control group; 46.2%(43 of 93) and 40%(83 of 206) in the group of patients and controls respectively, were heterozygous CT for MTHFR gene. The levels of homocysteine were 7.2 micromol/ml in the group of patients and 7.7 mmol/l in controls. There was no relationship between MTHFR gene polymorphisms and the increase of homocysteine levels, nor of these one with RPL. From the nutrigenetics perspective we suggest that studies related to MTHFR polymorphisms and the risk of disease include the levels of folate and B6 and B12 vitamins participating in the tetrahydrofolate cycle for trying to establish a direct relation among the genotype, the level of metabolite and the clinical manifestations. In this regard, we recommend the administration of folic acid in women in search of pregnancy due to the high frequency of heterozygous and homozygous for MTHFR C677T mutation in our population.

INTRODUCTION: Serum vitamin B12 concentration levels in children are essential to establish values in order to compare different regions or countries, and for considering e the possibility of supplementing diets with group B vitamins as a secondary prevention against cardiovascular diseases. MATERIAL AND METHODS: A cross-sectional epidemiological study was carried out to asses serum vitamin B12 levels in school children, 13-15 years of age, in Madrid. Folate and vitamin B12 vitamin determinations were performed on fasting blood samples. Genotype C677T of methylentetrahydrofolate reductase (MTHFR) enzyme was determined by PCR. RESULTS: The mean vitamin B12 level obtained in our study was 503 pmol/l; CI 95 % CI (478-528 pmol/l). The median was 471 pmol/l; interquartile range (IR)(337-632 pmol/l). No statistically significant differences were found by age or C677T genotype for MTHFR. Serum vitamin B12 concentrations were significantly higher in females. Prevalence of vitamin B12 deficiency (< 224 pmol/l) was 6 % in males and 4 % in females. CONCLUSIONS: Reference values for serum vitamin B12 concentrations in an adolescent population are presented. Prevalence of vitamin B12 deficiencies is higher in males.