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1.  Serum Lipid Profile in Subjects with Traumatic Spinal Cord Injury 
PLoS ONE  2015;10(2):e0115522.
Background and Aims
Few large studies have examined the relationship between spinal cord injury (SCI) and lipid profile. We studied serum lipid concentrations in subjects with traumatic SCI in relation to the degree of neurological involvement and time since injury, and compared them with values from a reference sample for the Spanish population (DRECE study).
Materials and Methods
A retrospective cohort was built from 177 consecutive cases with traumatic SCI admitted to the SCI unit of the Miguel Servet Hospital in Aragon (Spain). Outcome measures (cholesterol, triglycerides, HDL-c and LDL-c levels) were analyzed according to the ASIA Impairment Scale (AIS), neurological level of injury (involvement of all limbs vs. only lower limbs), and time since injury. All analyses were adjusted for age and sex.
Cases without preserved motor function (AIS A or B) had lower total and HDL cholesterol than the others (-11.4 [-21.5, -1.4] mg/dL total cholesterol and -5.1 [-8.8, -1.4] mg/dL HDL-c), and cases with all-limb involvement had lower total, HDL, and LDL cholesterol than those with only lower-limb involvement (-14.0 [-24.6, -3.4] mg/dL total cholesterol, -4.1 [-8.0, -0.2] mg/dL HDL-c, and -10.0 [-19.7, -0.3] mg/dL LDL-c) (all p<0.05). No association was found between lipid concentrations and time since injury. Concentrations of lipid subfractions and triglycerides in SCI subjects were lower than in sex- and age-stratified values from the reference sample.
A high degree of neurological involvement in SCI (anatomically higher lesions and AIS A or B) is associated with lower total cholesterol and HDL-c.
PMCID: PMC4338197  PMID: 25706982
2.  Association of Global DNA Methylation and Global DNA Hydroxymethylation with Metals and Other Exposures in Human Blood DNA Samples 
Environmental Health Perspectives  2014;122(9):946-954.
Background: The association between human blood DNA global methylation and global hydroxymethylation has not been evaluated in population-based studies. No studies have evaluated environmental determinants of global DNA hydroxymethylation, including exposure to metals.
Objective: We evaluated the association between global DNA methylation and global DNA hydroxymethylation in 48 Strong Heart Study participants for which selected metals had been measured in urine at baseline and DNA was available from 1989–1991 (visit 1) and 1998–1999 (visit 3).
Methods: We measured the percentage of 5-methylcytosine (5-mC) and 5-hydroxymethylcytosine (5-hmC) in samples using capture and detection antibodies followed by colorimetric quantification. We explored the association of participant characteristics (i.e., age, adiposity, smoking, and metal exposure) with both global DNA methylation and global DNA hydroxymethylation.
Results: The Spearman’s correlation coefficient for 5-mC and 5-hmC levels was 0.32 (p = 0.03) at visit 1 and 0.54 (p < 0.001) at visit 3. Trends for both epigenetic modifications were consistent across potential determinants. In cross-sectional analyses, the odds ratios of methylated and hydroxymethylated DNA were 1.56 (95% CI: 0.95, 2.57) and 1.76 (95% CI: 1.07, 2.88), respectively, for the comparison of participants above and below the median percentage of dimethylarsinate. The corresponding odds ratios were 1.64 (95% CI: 1.02, 2.65) and 1.16 (95% CI: 0.70, 1.94), respectively, for the comparison of participants above and below the median cadmium level. Arsenic exposure and metabolism were consistently associated with both epigenetic markers in cross-sectional and prospective analyses. The positive correlation of 5-mC and 5-hmC levels was confirmed in an independent study population.
Conclusions: Our findings support that both epigenetic measures are related at the population level. The consistent trends in the associations between these two epigenetic modifications and the characteristics evaluated, especially arsenic exposure and metabolism, suggest the need for understanding which of the two measures is a better biomarker for environmental epigenetic effects in future large-scale epidemiologic studies.
Citation: Tellez-Plaza M, Tang WY, Shang Y, Umans JG, Francesconi KA, Goessler W, Ledesma M, Leon M, Laclaustra M, Pollak J, Guallar E, Cole SA, Fallin MD, Navas-Acien A. 2014. Association of global DNA methylation and global DNA hydroxymethylation with metals and other exposures in human blood DNA samples. Environ Health Perspect 122:946–954;
PMCID: PMC4154208  PMID: 24769358
3.  Anopheles gambiae Croquemort SCRBQ2, expression profile in the mosquito and its potential interaction with the malaria parasite Plasmodium berghei 
The scavenger receptor family comprises transmembrane proteins involved in the recognition of polyanionic ligands. Several studies have established that members of this family are involved both in immunity and in developmental processes. In Drosophila melanogaster, one of the best characterized scavenger receptors is Croquemort, which participates in the recognition of apoptotic cells in the embryo. Although comparative genomic studies have revealed the presence of four orthologs of this receptor in the malaria vector Anopheles gambiae, little is known about their function. We have investigated the expression pattern of the four Croquemort orthologs during the mosquito life cycle. Croquemort transcripts SCRBQ2 and SCRBQ4 are expressed at all the developmental stages, while expression of Croquemort transcripts SCRBQ1 and SCRBQ3 is more restricted. We have also investigated the expression of the four Croquemort orthologs in the different organs of the adult female. Croquemort transcript SCRBQ2 is highly expressed in the A. gambiae female midgut. SCRBQ2 midgut gene expression was up-regulated after a non-infected or a Plasmodium berghei-infected blood meal, compared to its expression in midguts of sugar-fed females. Interestingly, knockdown of SCRBQ2 expression by dsRNA injection resulted in a 62.5% inhibition of oocyst formation, suggesting that SCRBQ2 plays a role in Plasmodium–mosquito interactions.
PMCID: PMC4138513  PMID: 19366631
Croquemort; Scavenger receptor; Anopheles; CD36; RNAi
4.  Selenium Status and Blood Lipids: The Cardiovascular Risk in Young Finns Study 
Journal of internal medicine  2011;270(5):469-477.
Concern has been recently raised about possible adverse cardio-metabolic effects of high selenium status, such as increased risks of diabetes and hyperlipidemia. However, most of the evidence comes from selenium-replete populations such as the US.
To examine cross-sectional and longitudinal associations of serum selenium with cardiovascular risk factors in Finland where selenium levels were among the lowest in the world until the early 1980s before the implementation of a nationwide selenium fertilization program.
Serum selenium was measured in 1,235 young Finns aged 3-18 years at baseline in 1980 (pre-fertilization), and in a subgroup (N=262) at the 6-year follow-up (1986, post-fertilization). During the 27-year follow-up, serum lipids, blood pressure, BMI, and smoking were assessed five times (1980, 1983, 1986, 2001, and 2007).
Mean (±SD) serum selenium concentrations were 74.3±14.0 ng/mL in 1980 and 106.6±12.5 ng/mL in 1986 (average increase 32.3 ng/mL; 95% CI: 30.3 to 34.3, p<0.0001). In univariate and multivariable cross-sectional models in 1980 and 1986, increased serum selenium levels were consistently associated with increased total, HDL- and LDL-cholesterol. However, the average longitudinal changes in lipids were −0.20 mmol/L (95% CI: −0.30 to −0.10, p<0.0001) for total cholesterol, 0.06 mmol/L (95% CI: 0.03 to 0.10, p<0.0001) for HDL-cholesterol, and −0.23 mmol/L (95% CI: −0.31 to −0.14, p<0.0001) for LDL-cholesterol. Selenium measured in 1986 was not associated with lipids assessed in 2001 and 2007.
Cross-sectional findings from the Young Finns study corroborate positive associations of selenium status with serum lipids. However, longitudinal evidence does not support the causality of this link.
PMCID: PMC3172343  PMID: 21554435
cardiovascular; cross-sectional; follow-up; lipids; risk factors; selenium
5.  Aragon workers’ health study – design and cohort description 
Spain, a Mediterranean country with relatively low rates of coronary heart disease, has a high prevalence of traditional cardiovascular risk factors and is experiencing a severe epidemic of overweight/obesity. We designed the Aragon Workers’ Health Study (AWHS) to characterize the factors associated with metabolic abnormalities and subclinical atherosclerosis in a middle aged population in Spain free of clinical cardiovascular disease. The objective of this paper is to describe the study design, aims and baseline characteristics of participants in the AWHS.
Longitudinal cohort study based on the annual health exams of 5,400 workers of a car assembly plant in Figueruelas (Zaragoza, Spain). Study participants were recruited during a standardized clinical exam in 2009–2010 (participation rate 95.6%). Study participants will undergo annual clinical exams and laboratory assays, and baseline and triennial collection of biological materials for biobanking and cardiovascular imaging exams (carotid, femoral and abdominal ultrasonography, coronary calcium score, and ankle-arm blood pressure index). Participants will be followed-up for 10 years.
The average (SD) age, body mass index, and waist circumference were 49.3 (8.7) years, 27.7 (3.6) kg/m2 and 97.2 (9.9) cm, respectively, among males (N = 5,048), and 40.8 (11.6) years, 24.4 (3.8) kg/m2, and 81.9 (9.9) cm, among females (N = 351). The prevalence of overweight, obesity, current smoking, hypertension, hypercholesterolemia, and diabetes were 55.0, 23.1, 37.1, 40.3, 75.0, and 7.4%, respectively, among males, and 23.7, 8.3, 45.0, 12.1, 59.5, and 0.6%, respectively, among females. In the initial 587 study participants who completed all imaging exams (94.5% male), the prevalence of carotid plaque, femoral plaque, coronary calcium score >1 to 100, and coronary calcium score >100 was 30.3, 56.9, 27.0, and 8.8%, respectively. 67.7% of study participants had at least one plaque in the carotid or femoral arteries.
Baseline data from the AWHS show a high prevalence of cardiovascular risk factors and of sublinical atherosclerosis. Follow-up of this cohort will allow the assessment of subclinical atherosclerosis progression and the link of disease progression to traditional and emergent risk factors.
PMCID: PMC3439398  PMID: 22712826
6.  Serum selenium and serum lipids in US adults: National Health and Nutrition Examination Survey (NHANES) 2003–2004 
Atherosclerosis  2010;210(2):643-648.
High selenium has been recently associated with several cardiovascular and metabolic risk factors including diabetes, blood pressure and lipid levels. We evaluated the association of serum selenium with fasting serum lipid levels in the National Health and Nutrition Examination Survey (NHANES) 2003–2004, the most recently available representative sample of the US population that measured selenium levels.
Cross-sectional analysis of 1159 adults ≥40 years old from NHANES 2003–2004. Serum selenium was measured by inductively coupled plasma-dynamic reaction cell-mass spectrometry. Fasting serum total-cholesterol, triglycerides, and HDL cholesterol were measured enzymatically and LDL cholesterol was calculated.
Mean serum selenium was 136.7 µg/L. The multivariable adjusted average differences (95% confidence interval) comparing the highest (≥147 µg/L) to the lowest (<124 µg/L) selenium quartiles were 18.9 (9.9, 28.0) mg/dL for total cholesterol, 12.7 (3.3, 22.2) mg/dL for LDL cholesterol, 3.9 (0.4, 7.5) mg/dL for HDL cholesterol, and 11.5 (−7.6, 30.7) mg/dL for triglycerides. In spline regression models, total and LDL cholesterol levels increased progressively with increasing selenium concentrations. HDL cholesterol increased with selenium but reached a plateau above 120 µg/L of serum selenium (20th percentile). The triglyceride-selenium relationship was U-shaped.
In US adults, high serum selenium concentrations were associated with increased serum concentrations of total and LDL cholesterol. Selenium was associated with increasing HDL cholesterol only at low selenium levels. Given increasing trends in dietary selenium intake and supplementation, the causal mechanisms underlying these associations need to be fully characterized.
PMCID: PMC2878899  PMID: 20102763
Selenium; Serum Lipids; NHANES
7.  A prospective study of dietary selenium intake and risk of type 2 diabetes 
BMC Public Health  2010;10:564.
Growing evidence raises concern about possible associations of high selenium exposure with diabetes in selenium-replete populations such as the US. In countries with lower selenium status, such as Italy, there is little epidemiological evidence on the association between selenium and diabetes. This study examined the prospective association between dietary selenium intake and risk of type 2 diabetes.
The ORDET cohort study comprised a large sample of women from Northern Italy (n = 7,182). Incident type 2 diabetes was defined as a self-report of a physician diagnosis, use of antidiabetic medication, or a hospitalization discharge. Dietary selenium intake was measured by a semi-quantitative food-frequency questionnaire at the baseline examination (1987-1992). Participants were divided in quintiles based on their baseline dietary selenium intake.
Average selenium intake at baseline was 55.7 μg/day. After a median follow-up of 16 years, 253 women developed diabetes. In multivariate logistic regression analyses, the odds ratio for diabetes comparing the highest to the lowest quintile of selenium intake was 2.39, (95% CI: 1.32, 4.32; P for linear trend = 0.005). The odds ratio for diabetes associated with a 10 μg/d increase in selenium intake was 1.29 (95% CI: 1.10, 1.52).
In this population, increased dietary selenium intake was associated with an increased risk of type 2 diabetes. These findings raise additional concerns about the association of selenium intake above the Recommended Dietary Allowance (55 μg/day) with diabetes risk.
PMCID: PMC2949772  PMID: 20858268
8.  Serum selenium concentrations and hypertension in the US population 
Selenium is an antioxidant micronutrient with potential interest for cardiovascular disease prevention. Few studies have evaluated the association between selenium and hypertension, with inconsistent findings. We explored the relationship of serum selenium concentrations with blood pressure and hypertension in a representative sample of the US population.
Methods and Results
Cross-sectional analysis of 2,638 adults ≥40 year old who participated in the National Health and Nutrition Examination Survey (NHANES) 2003–2004. Serum selenium was measured by inductively coupled plasma-dynamic reaction cell-mass spectrometry. Hypertension was defined as blood pressure ≥140/90 mmHg or current use of antihypertensive medication. Mean serum selenium was 137.1 µg/L. The multivariable adjusted differences (95% confidence interval) in blood pressure levels comparing the highest (≥150 µg/L) to the lowest (<122 µg/L) quintile of serum selenium were 4.3 (1.3, 7.4), 1.6 (−0.5, 3.7) and 2.8 (0.8, 4.7) mmHg for systolic, diastolic, and pulse pressure, respectively. The corresponding odds ratio (95% CI) for hypertension was 1.73 (1.18, 2.53). In spline regression models, blood pressure levels and the prevalence of hypertension increased with increasing selenium concentrations up to 160 µg/L.
High serum selenium concentrations were associated with higher prevalence of hypertension. These findings call for a thorough evaluation of the risks and benefits associated with high selenium status in the US.
PMCID: PMC2773506  PMID: 20031863
Selenium; blood pressure; hypertension; National Health and Nutrition Examination Survey; NHANES
9.  Serum Selenium and Peripheral Arterial Disease: Results From the National Health and Nutrition Examination Survey, 2003–2004 
American Journal of Epidemiology  2009;169(8):996-1003.
The authors conducted a cross-sectional study of the association of serum selenium with the prevalence of peripheral arterial disease among 2,062 US men and women 40 years of age or older participating in the National Health and Nutrition Examination Survey, 2003–2004. Serum selenium was measured by using inductively coupled plasma-dynamic reaction cell-mass spectrometry. Peripheral arterial disease was defined as an ankle-brachial blood pressure index <0.90. The age-, sex-, and race-adjusted prevalence of peripheral arterial disease decreased with increasing serum selenium (P for linear trend = 0.02), but there was an indication of an upturn in risk in the highest quartile of serum selenium. The fully adjusted odds ratios for peripheral arterial disease comparing selenium quartiles 2, 3, and 4 with the lowest quartile were 0.75 (95% confidence interval: 0.37, 1.52), 0.58 (95% confidence interval: 0.28, 1.19), and 0.67 (95% confidence interval: 0.34, 1.31), respectively. In spline regression models, peripheral arterial disease prevalence decreased with increasing serum selenium levels up to 150–160 ng/mL, followed by a gradual increase at higher selenium levels. The association between serum selenium levels and the prevalence of peripheral arterial disease was not statistically significant, although a U-shaped relation was suggested.
PMCID: PMC2727225  PMID: 19221120
antioxidants; cardiovascular diseases; cross-sectional studies; nutrition surveys; peripheral vascular diseases; selenium
10.  Physical inactivity interacts with an endothelial lipase polymorphism to modulate high density lipoprotein cholesterol in the GOLDN study 
Atherosclerosis  2009;206(2):500-504.
Plasma high density lipoprotein (HDL) cholesterol (HDL-C) concentration is highly heritable but is also modifiable by environmental factors including physical activity. HDL-C response to exercise varies among individuals, and this variability may be associated with genetic polymorphisms in the key regulators of HDL metabolism including endothelial lipase (LIPG).
We examined associations between variants LIPG T111I (rs2000813) and LIPG i24582 (rs6507931), HDL and television viewing/computer use (“screen time”) as a marker for physical inactivity in a population with high prevalence of metabolic syndrome. Subjects consisted of 539 White men and 584 women (mean ± S.D., 49 ± 16 years) participating in the GOLDN study.
We did not observe an association with either LIPG SNP or HDL independently of screen time. In multi-adjusted linear regression models, HDL interacted significantly with screen time as a continuous variable in LIPG i24582 subjects with TT genotype (P < 0.05). By dichotomizing screen time into high and low levels, we found significant genotype-associated differences in HDL in women but not men. When screen time was ≥2.6 h/day, the concentrations of total HDL-C, large HDL, large low density lipoprotein (LDL) were lower, the concentration of small LDL was higher and HDL and LDL particle sizes were smaller in subjects with LIPG i24582 TT compared to CT and CC subjects (P < 0.05).
We found a significant gene-physical inactivity interaction for HDL and some LDL measures for the LIPG i24582 polymorphism. Higher levels of physical activity may be protective for HDL-C concentrations and low activity detrimental in LIPG i24582 TT individuals, especially in women.
PMCID: PMC2801595  PMID: 19380136
LIPG; Endothelial lipase; HDL; Television; Physical activity
11.  Serum Selenium Concentrations and Diabetes in U.S. Adults: National Health and Nutrition Examination Survey (NHANES) 2003–2004 
Environmental Health Perspectives  2009;117(9):1409-1413.
Increasing evidence suggests that high selenium levels are associated with diabetes and other cardiometabolic risk factors.
We evaluated the association of serum selenium concentrations with fasting plasma glucose, glycosylated hemoglobin levels, and diabetes in the most recently available representative sample of the U.S. population.
We used a cross-sectional analysis of 917 adults ≥ 40 years of age who had a fasting morning blood sample in the National Health and Nutrition Examination Survey 2003–2004. We evaluated the association of serum selenium, measured by inductively coupled plasma-dynamic reaction cell-mass spectrometry, and diabetes, defined as a self-report of current use of hypoglycemic agents or insulin or as fasting plasma glucose ≥ 126 mg/dL.
Mean serum selenium was 137.1 μg/L. The multivariable adjusted odds ratio [95% confidence interval (CI)] for diabetes comparing the highest quartile of serum selenium (≥ 147 μg/L) with the lowest (< 124 μg/L) was 7.64 (3.34–17.46). The corresponding average differences (95% CI) in fasting plasma glucose and glycosylated hemoglobin were 9.5 mg/dL (3.4–15.6 mg/dL) and 0.30% (0.14–0.46%), respectively. In spline regression models, the prevalence of diabetes as well as glucose and glycosylated hemoglobin levels increased with increasing selenium concentrations up to 160 μg/L.
In U.S. adults, high serum selenium concentrations were associated with higher prevalence of diabetes and higher fasting plasma glucose and glycosylated hemoglobin levels. Given high selenium intake in the U.S. population, further research is needed to determine the role of excess selenium levels in the development or the progression of diabetes.
PMCID: PMC2737018  PMID: 19750106
diabetes; glycosylated hemoglobin; National Health and Nutrition Examination Survey; NHANES; selenium

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