Folate nutritional status and the 677C>;T polymorphism of methylene tetrahydrofolate reductase (MTHFR) are known to influence gene expression. The objective of this study was to determine if supplementation with 400μg of folic acid for a period of 12 weeks will change gene expression in individuals with low folate status. We also determined the influence of MTHFR 677C>;T on these outcomes. 20 men homozygous for the variant allele of MTHFR and 20 men without the polymorphism, aged 45–65, whose average plasma folate was 10.53 nmol/L were selected for the study from Milan, Italy. The subjects were randomized to receive placebo or 400μg folic acid tablets. Blood samples were collected before and after 12 weeks of supplementation. When compared to subjects on placebo, those who received folic acid had higher folate concentration (9.78 nmol/L vs 44.56 nmol/L, P<0.0001) and lower homocysteine concentration (12.59 μmol/L vs 9.05 μmol/L, P=0.1) in plasma at the end of 12 weeks. Changes in gene expression in leukocytes were determined by microarray analysis. Expression of multiple genes for pro‐inflammatory chemokines and interleukins decreased with folic acid supplementation, especially in TT genotypes of MTHFR. Folic acid supplementation at current RDA can potentially alter inflammation in individuals with low folate status. Supported by USDA grant 2005–35200‐15296 & USDA cooperative agreement 51520–008‐04S.

Introduction Treatment with B-vitamins and betaine reduces the high risk of thrombosis in patients with homocystinuria, a metabolic syndrome that is characterized by severe hyperhomocysteinemia (HHcy). In contrast, there is no clear demonstration that B-vitamins reduce the risk of thrombosis in patients with mild HHcy: for this reason, many question the clinical utility of measuring total Hcy (tHcy) in patients with thrombosis. However, thrombosis may be the first clinical manifestation of homocystinuria in patients reaching adulthood without signs and symptoms of the syndrome. Aim 1) to measure the prevalence of severe, previously undiagnosed, HHcy among patients with thrombosis 2) to profile these patients on the basis of their characteristics. Methods Six Italian Thrombosis Centers completed a first questionnaire, reporting tHcy levels in patients with thrombosis who underwent thrombophilia screening, and a second questionnaire, reporting the characteristics of patients with severe HHcy (tHcy>100μmol/L). Results Of 19,678 cross-sectionally collected patients with thrombosis who underwent thrombophilia screening in the last 12.5years (median value, range 6-17), 38 had severe HHcy (0.2%). Their median age at diagnosis was 47years (range 19-83) and the median level of tHcy was 130μmol/L (range 101-262). Venous thromboembolism (71%) was more frequent than arterial thromboembolism (26%); recurrent thrombosis occurred in 42% of cases. Conclusions Measurement of tHcy in adult patients with thrombosis may reveal the presence of severe HHcy. Since treatment of patients with severe HHcy decreases the risk of thrombosis, measurement of tHcy in patients with thrombosis may prove clinically useful.

Background: methionine ingestion (100 mg/kg) identifies subjects in whom fasting total homocysteine (tHcy) may be normal but the post-methionine load (PML) tHcy is abnormally high.Methods: In 96 subjects [ 54 M/42 F, 40.4 +/- 12.3 yrs old; 28 with the 68 bp844 ins of the Cystathionine-beta-synthase gene (CBSins+); 20 homozygotes for the C677T mutation of the methylene-tetrahydrofolate reductase gene (MTHFR++); 13 with the combination of the two, and 35 without any of them], we have evaluated in vivo oxidative stress and platelet activation, as reflected by urinary excretions of 8-iso-PGF2 alpha and of 11-dehydro-TXB(2) respectively, before and after a methionine load test (PML). A history of early-onset thrombosis (18 arterial, 32 venous, 2 both) was present in 52/96 of them.Results: Baseline; tHcy was highest in MTHFR++ carriers (p < 0,05); 8-iso-PGF2 alpha and 11-dehydro-TXB(2) levels were independent of sex, MTHFR++ and/or CBSins+(p > 0.05). PML; The similar to 3-fold increase (p < 0.01 vs baseline) in tHcy reached a plateau within 6-8 hrs. Mean PML tHcy was maximal in MTHFR++ carriers (p= 0.000). 8-iso-PGF2 alpha and 11-dehydro-TXB(2) increase reached a maximum within 4 hrs. 11-dehydro-TXB2 increase was highest (p= 0.023 vs baseline) in subjects with a history of thrombosis. Baseline 11-dehydro-TXB(2) and a history of thrombosis independently predicted PML 11-dehydro-TXB2 (beta= 0.287, p= 0.000 and beta= 0.308, p= 0.026, respectively). The PML increase in 8-iso-PGF2 alpha or in 11-dehydro-TXB2 were comparable in the different genotypes (p > 0.05).Conclusion: regardless genotypes associated with moderate hyperhomocysteinemia, following a methionine loading test, in vivo oxidative stress and platelet activation occur, being the latter maximal in subjects with a history of early-onset thrombosis. (C) 2011 Elsevier Ltd. All rights reserved.

Vitamin B6 participates as a coenzyme to >100 reactions and is required for the degradation of homocysteine, a strong predictor of atherothrombotic events. Low circulating vitamin B6, however, predisposes to arterial and venous thrombosis, independently of hyperhomocysteinaemia [ [1] Vanuzzo D. Pilotto L. Lombardi R. Lazzerini G. Carluccio M. Diviacco S. et al. Both vitamin B6 and total homocysteine plasma levels predict long-term atherothrombotic events in healthy subjects. Eur Heart J. 2007; 28: 484-491 Crossref PubMed Scopus (31) Google Scholar ]. Populations studies suggest an intriguing link between low vitamin B6 and systemic chronic inflammation, presumably because vitamin B6 is consumed during inflammation [ [2] Morris M.S. Sakakeeny L. Jacques P.F. Picciano M.F. Selhub J. Vitamin B6 intake is inversely related to, and the requirement is affected by, inflammatory status. J Nutr. 2010; 140: 103-110 Crossref PubMed Scopus (87) Google Scholar ]. Chronic Obstructive Pulmonary Disease (COPD) is a long-lasting systemic inflammatory illness and confers an independent higher risk of atherothrombotic events [ [3] Johnston A.K. Mannino D.M. Hagan G.W. Davis K.J. Kiri V.A. Relationship between lung function impairment and incidence or recurrence of cardiovascular events in a middle-aged cohort. Thorax. 2008; 63: 599-605 Crossref PubMed Scopus (127) Google Scholar ]. While investigating hyperhomocysteinaemia in COPD, we have described for the first time lower vitamin B6 plasma concentrations in COPD patients than in matched controls [ [4] Fimognari F.L. Loffredo L. Di Simone S. Sampietro F. Pastorelli R. Monaldo M. et al. Hyperhomocysteinaemia and poor vitamin B status in chronic obstructive pulmonary disease. Nutr Metab Cardiovasc Dis. 2009; 19: 654-659 Abstract Full Text Full Text PDF PubMed Scopus (30) Google Scholar ]. We have now re-analyzed our study sample to determine the mechanisms accounting for this COPD-related poor vitamin B6 status.

Human chromosomes are capped by telomeres, which consist of tandem repeats of DNA and associated proteins. The length of the telomeres is reduced with increasing cell divisions except when the enzyme telomerase is active, as in stem cells and germ cells. Telomere dysfunction has been associated with development of age-related pathologies, including cancer, cardiovascular disease, Alzheimer's disease, and Parkinson's disease. DNA damage in the telomeric region causes attrition of telomeres. Because folate provides precursors for nucleotide synthesis and thus affects the integrity of DNA, including that of the telomeric region, folate status has the potential to influence telomere length. Telomere length is epigenetically regulated by DNA methylation, which in turn could be modulated by folate status. In this study, we determined whether folate status and the 677C > T polymorphism of the methylene tetrahydrofolate reductase (MTHFR) gene are associated with the telomere length of peripheral blood mononuclear cells in healthy men. The results of our study showed that plasma concentration of folate was associated with telomere length of peripheral blood mononuclear cells in a nonlinear manner. When plasma folate concentration was above the median, there was a positive relationship between folate and telomere length. In contrast, there was an inverse relationship between folate and telomere length when plasma folate concentration was below the median. The MTHFR 677C > T polymorphism was weakly associated (P = 0.065) with increased telomere length at below-median folate status. We propose that folate status influences telomere length by affecting DNA integrity and the epigenetic regulation of telomere length through DNA methylation.

PURPOSE:To evaluate the prevalence of moderate fasting hyperhomocysteinemia (HHcy) and postmethionine load (PML) HHcy among patients with early-onset central retinal vein occlusion (CRVO). METHODS:The prevalence of fasting HHcy and that of PML HHcy were evaluated in a consecutive series of 58 patients with CRVO who were younger than 56 years of age (mean age, 40.3 years) and in 103 controls (mean age, 39.6 years). Plasma folate, vitamin B12, and pyridoxal-5'-phosphate (PLP) levels were measured in 42 patients and 67 controls. RESULTS:Mantel-Haenszel odds ratios for CRVO patients were 3.00 (95% confidence interval [CI], 0.83-10.8) for fasting HHcy, 3.50 (95% CI, 1.07-11.4) for PML HHcy, and 3.00 (1.18-7.6) for fasting HHcy and PML HHcy in subjects with normal fasting total homocysteine (tHcy) levels. Moderate HHcy was associated with reduced plasma levels of folate and PLP (P < or = 0.04). There was no significant dependence of fasting and PML tHcy levels on any traditional risk factor evaluated. CONCLUSIONS:Moderate HHcy is an independent risk factor for early-onset CRVO.

BACKGROUND AND OBJECTIVES High lipoprotein a [Lp(a)] and fibrinogen levels are suggested risk factors for coronary heart disease (CHD) and stroke morbidity and mortality. Experimental data strongly suggest that the mechanisms of atherothrombosis include an interaction between fibrinogen and Lp(a), but little clinical evidence of a synergism between these two parameters has been reported. DESIGN AND METHODS Within the frame of a prospective population study conducted in the area of Cremona (Lombardy, Italy), 343 women and 216 men aged > or =65 years were evaluated for clinical and biochemical cardiovascular risk factors. Lp(a) levels > or =30 mg/dL were observed in 22.7% and 23.9% of men and women, respectively. Fibrinogen levels were higher in women (p<0.0001). After a median follow-up of 6.3 years 107 deaths were recorded, of which 33 were due to CHD or ischemic stroke. RESULTS The combined incidence rate of CHD and stroke mortality increased from 10.8 (per 1000 person-years) for subjects with either Lp(a) > or =30 mg/dL or fibrinogen within the 5th quintile of the gender-specific distribution to 38.4 for subjects with both Lp(a) > or =30 mg/dL and fibrinogen within the 5th quintile. Age (p<0.0001), insulin (p<0.0002) and the combination of high Lp(a) and fibrinogen (hazard ratio=3.11, p=0.014), but not fibrinogen or Lp(a) levels in isolation, were independent predictors of CHD and stroke mortality. In a subgroup of 447 subjects in whom C-reactive protein (CRP) was measured, CRP levels were not predictive of combined CHD and stroke mortality. INTERPRETATIONS AND CONCLUSIONS Based on these results obtained in a relatively small population of elderly subjects, the association of high Lp(a) and fibrinogen levels appears to carry an increased risk of pooled CHD and stroke mortality.

BACKGROUND AND OBJECTIVES:Atrial fibrillation is complicated by a high rate of ischemic stroke. Previous studies have shown that an increased level of circulating total plasma homocysteine (tHcy) is an independent predictor of stroke, but it is unclear whether it is also predictive of stroke in patients with atrial fibrillation. The objective of this study was to evaluate whether increased tHcy is an independent predictor of cardio-embolic stroke in patients with non-valvular atrial fibrillation. DESIGN AND METHODS:We studied 163 consecutive patients (77 males and 86 females; mean age 72.3+/-8.8 years) with permanent (n=118) or paroxysmal (n=45) atrial fibrillation of non-valvular origin hospitalized for cardiac reasons. Ischemic stroke, documented by nuclear magnetic resonance or computerized tomography imaging, had occurred at an average of 2 years before hospitalization in 40 patients (16 males and 24 females, mean age 74.8+/-8.8 years). Fasting tHcy levels were determined by high performance liquid chromatography. RESULTS:Multivariate analysis adjusting for traditional cardiovascular risk factors, thromboembolic risk factors and predictors of tHcy (glomerular filtration rate, uric acid, gender) and fibrinogen levels (age, alcohol intake) showed that total homocysteine (OR: 1.056; for each 1 micromol/L increase, 95% C.I.: 1.00-1.12; p=0.042) and fibrinogen (OR: 1.008 for each 1 mg/dL increase; 95% C.I.: 1.00-1.014; p=0.016) were independently associated with ischemic stroke. With respect to patients in the first quartile of the tHcy distribution (4.6-7.5 micromol/L), patients in the fourth quartile of the tHcy distribution (18.7-67.1 micromol/L) had a 2.73-fold increased probability of ischemic stroke INTERPRETATION AND CONCLUSIONS:In patients with non-valvular atrial fibrillation hospitalized for cardiac reasons, increased fasting tHcy levels are independently associated with a history of ischemic stroke.

The authors demonstrate in vitro antiviral activity of a methanolic extract of Combretum micranthum leaves against HSV-1 and HSV-2. This activity is present only in the extract dissolved 7 days before the assay, but not in the freshly prepared extract, thus indicating the presence of inactive precursors which undergo spontaneous transformations into active compounds. The alkaline autooxidation of the methanolic extract promotes this rapid transformation. The precursors have been identified as condensed catechinic tannins, which, under alkaline conditions, suffer rapid cleavage, intramolecular rearrangement to catechinic acid and autooxidation. The alkaline autooxidation products of the methanolic extract of C. micranthum and those of the synthetic catechinic acid show similar I.R. and U.V. absorption curves, as well as similar anti-HSV-1 and -HSV-2 activities. EC50s of catechinic acid autooxidation products against HSV-1 and HSV-2 replication were 2 μg/ml and 4 μg/ml, respectively, when cell cultures were treated with the compound during virus infection.