Circulating microRNA in cardiovascular disease.

Acute myocardial infarction (AMI) and heart failure (HF) are two major causes of cardiovascular mortality and morbidity. Early diagnosis of these conditions is essential to instigate immediate treatment that may result in improved outcomes. Traditional biomarkers of AMI include cardiac troponins and other proteins released from the injured myocardium but there are a number of limitations with these biomarkers especially with regard to specificity. In the past few years circulating nucleic acids, notably microRNA that are small non-coding RNAs that regulate various cellular processes, have been investigated as biomarkers of disease offering improved sensitivity and specificity in the diagnosis and prognostication of various conditions. In this review, the role of microRNAs as biomarkers used in the diagnosis of AMI and HF is discussed, their advantage over traditional biomarkers is outlined and the potential for their implementation in clinical practice is critically assessed.

Plasma insulin-like factor 3 (INSL3) in male patients with osteoporosis and Klinefelter's syndrome.

Insulin-like factor 3 (INSL3) is a peptide hormone produced in leydig cells of the testes. Its role in the adult male is unknown but INSL3 and its receptor RXFP2 have been linked to bone cell differentiation. It is speculated that low levels of INSL3 could be responsible for low bone mineral density in patients with primary osteoporosis and Klinefelter's Syndrome. The aim of this study was to assess plasma INSL3 in patients with osteoporosis and Klinefelter's Syndrome compared to healthy males. Fourteen healthy males, 21 males with osteoporosis (4 primary and 17 secondary) and 4 patients with Klinefelter's Syndrome were studied. Plasma INSL3, testosterone, LH, FSH and Sex hormone-binding globulin were evaluated. Plasma INSL3 concentrations were similar in osteoporosis patients compared to healthy controls (0.72 vs. 0.69 ng/mL, p = 0.26). INSL3 was significantly higher in patients with primary osteoporosis (n = 4) compared to age-matched healthy controls (n = 8) (0.845 vs. 0.665 ng/mL, p = 0.021). INSL3 levels in Klinefelter's Syndrome patients were significantly lower compared to healthy controls (0.39 vs. 0.69 ng/mL, p = 0.01). Plasma INSL3 levels were lower in Klinefelter's Syndrome reflecting testicular failure. INSL3 levels were not lower in men with osteoporosis. The relationship between INSL3, its receptor and bone metabolism requires further study.

Changes in the concentrations of creatinine, cystatin C and NGAL in patients with acute paraquat self-poisoning.

An increase in creatinine > 3 µmol/L/h has been suggested to predict death in patients with paraquat self-poisoning and the value of other plasma biomarkers of acute kidney injury has not been assessed. The aim of this study was to validate the predictive value of serial creatinine concentrations and to study the utility of cystatin C and neutrophil gelatinase-associated lipocalin (NGAL) as predictors of outcome in patients with acute paraquat poisoning. The rate of change of creatinine (dCr/dt) and cystatin C (dCyC/dt) concentrations were compared between survivors and deaths. Receiver-operating characteristic (ROC) curves were constructed to determine the best threshold for predicting death. Paraquat was detected in 20 patients and 7 of these died between 18 h and 20 days post-ingestion. The dCr/dt ROC curve had an area of 0.93 and the cut-off was > 4.3 µmol/L/h (sensitivity 100%, specificity 85%, likelihood ratio 7). The dCyC/dt ROC curve had an area of 0.97 and the cutoff was > 0.009 mg/L/h (sensitivity 100%, specificity 91%, likelihood ratio 11). NGAL did not separate survivors from deaths. Death due to acute paraquat poisoning is associated with changes in creatinine and cystatin concentrations. Further validation of these measurements is needed before they can be adopted in guiding intensive treatments.

Pigmentation and vitamin D metabolism in Caucasians: low vitamin D serum levels in fair skin types in the UK.

BACKGROUND: Vitamin D may play a protective role in many diseases. Public health messages are advocating sun avoidance to reduce skin cancer risk but the potential deleterious effects of these recommendations for vitamin D metabolism have been poorly investigated. METHODOLOGY/PRINCIPAL FINDINGS: We investigated the association between 25-hydroxy-vitamin D (25(OH)D), skin type and ultraviolet exposure in 1414 Caucasian females in the UK. Mean age of the cohort was 47 years (18-79) and mean 25(OH)D levels were 77 nmol/L (6-289). 25(OH)D levels were strongly associated with season of sampling with higher levels in the spring and summer months (p<0.0001). Light skin types (skin type 1 and 2) have lower levels of 25(OH)D (mean 71 nmol/L) compared to darker skin types (skin type 3 and 4) (mean 82 nmol/L) after adjusting for multiple confounders (p<0.0001). The trend for increasing risk of low vitamin D with fairer skin types was highly significant despite adjustment for all confounders (p = 0.001). CONCLUSIONS/SIGNIFICANCE: Contrary to previous studies across different ethnic backgrounds, this study within Caucasian UK females shows that fair skin types have lower levels of 25(OH)D compared to darker skin types with potential detrimental health effects. Public health campaigns advocating sun avoidance in fair skinned individuals may need to be revised in view of their risk of vitamin D deficiency.

An obesogenic postnatal environment is more important than the fetal environment for the development of adult adiposity: a study of female twins.

BACKGROUND: A relation between birth weight and adult body composition has been reported in singleton populations, especially when more accurate measures of body composition, such as dual-energy X-ray absorptiometry (DXA) were used. It remains uncertain whether this is mediated by a direct effect of fetal nutrition, through factors in the shared environment, or through genetic factors. OBJECTIVE: The objective was to investigate the relation between birth weight and body composition with the use of a co-twin design. DESIGN: DXA measurements and birth weights were available for 2228 dizygotic and 842 monozygotic female twins aged between 18 and 80 y. Multivariate regression models were used to identify both individual specific relations and those mediated through the shared environment. RESULTS: Significant relations were found between birth weight and DXA measures for individuals. A 1-kg increase in birth weight was associated with a 1.72-kg increase in lean mass, a 0.25-kg increase in fat mass, and a 0.05-unit increase in the lean:fat mass ratio. Within twin pairs, the analysis showed that associations between birth weight and absolute levels of lean and fat mass were mediated through individual-specific effects, whereas the relation between birth weight and the proportion of lean to fat mass was mediated purely through factors common to twin pairs. CONCLUSIONS: A higher birth weight is associated with a higher proportion of lean to fat mass as adults. However, these analyses suggest that this association is not determined by individual specific factors in utero (eg, fetal nutrition) but through factors in the shared common environment of the twins.

Epidemiology and genetic epidemiology of the liver function test proteins.

BACKGROUND: The liver function test (LFT) is among the most commonly used clinical investigations to assess hepatic function, severity of liver diseases and the effect of therapies, as well as to detect drug-induced liver injury (DILI). AIMS: To determine the relative contribution of genetic and environmental factors as well as test and quantify the effects of sex, age, BMI and alcohol consumption to variation in liver function test proteins--including alanine amino transaminase (ALT), Albumin, gamma glutamyl transpeptidase (GGT), total bilirubin, total protein, total globulin, aspartate transaminase (AST), and alkaline phosphotase (ALP)--using the classical twin model. METHODS: Blood samples were collected from a total of 5380 twin pairs from the TwinsUK registry. We measured the expression levels of major proteins associated with the LFT, calculated BMI from measured weight and height and questionnaires were completed for alcohol consumption by the twins. The relative contribution of genetic and environmental factors to variation in the LFT proteins was assessed and quantified using a variance components model fitting approach. RESULTS: Our results show that (1) variation in all the LFTs has a significant heritable basis (h(2) ranging from 20% to 77%); (2) other than GGT, the LFTs are all affected to some extent by common environmental factors (c(2) ranging from 24% to 54%); and (3) a small but significant proportion of the variation in the LFTs was due to confounding effects of age, sex, BMI, and alcohol use. CONCLUSIONS: Variation in the LFT proteins is under significant genetic and common environmental control although sex, alcohol use, age and BMI also contribute significantly to inter-individual variation in the LFT proteins. Understanding the underlying genetic contribution of liver function tests may help the interpretation of their results and explain wide variation among individuals.

CD36 gene promoter polymorphisms are associated with low density lipoprotein-cholesterol in normal twins and after a low-calorie diet in obese subjects.

Common polymorphisms of the CD36 fatty acid transporter gene have been associated with lipid metabolism and cardiovascular disease. Association of a CD36 promoter single nucleotide polymorphism genotype with anthropometry and serum lipids was investigated in normal subjects, and in obese subjects during an 8-week low calorie diet and 6-month weight-maintenance period. 2728 normal female Twins UK subjects (mean body mass index 24.8 +/- 4.4 kg/m2; age 47.3 +/- 12.5 y) and 183 obese male and female Spanish subjects (mean body mass index 30.6 +/- 3.0 kg/m2; age 35.0 +/- 5.0 y) were genotyped for the CD36-22674 T/C (rs2151916) promoter single nucleotide polymorphism. In the Twins UK full cohort, the C-allele was associated with lower low density lipoprotein-cholesterol (p = .02, N = 2396). No associations were found in the obese Spanish subjects at baseline, but 6 months after the end of the low-calorie diet, the C-allele was associated with lower total- (p = .03) and low density lipoprotein-cholesterol (p = .01) and higher high density lipoprotein-cholesterol (p = .01). Intake of saturated fatty acids was lower in carriers of the C-allele at baseline, but not significantly so (p = .11). However, 6 months after the end of the low-calorie diet, elements of the lipid profile were correlated with saturated fatty acid intake: total cholesterol r = .21, p = .060; low density lipoprotein-cholesterol: r = .25, p = .043; high density lipoprotein-cholesterol: r = -.26, p = .007. CD36 promoter SNP allele -22674C is therefore associated with lower serum low-density lipoprotein-cholesterol in normal female twins and with improved lipid profile during weight loss and maintenance in obese subjects.

Adiponectin gene ADIPOQ SNP associations with serum adiponectin in two female populations and effects of SNPs on promoter activity.

Adiponectin is an insulin sensitiser in muscle and liver, and low serum levels characterise obesity and insulin resistance. Eight tagging single nucleotide polymorphisms (tSNPs) in the ADIPOQ gene and promoter were selected, and association with serum adiponectin was tested, in two independent samples of Caucasian women: the Chingford Study (n = 808, mean age 62.8 +/- 5.9 years) and Twins UK (n = 2,718, mean age 47.4 +/- 12.6 years). In the Chingford cohort, -11391 G/A, -10066 G/A (rs182052), -7734 C/A (rs16861209), +276 G/T (rs1501299) and +3228 C/T (rs1063537) were significantly associated with fasting serum adiponectin (Ps = 1.00 x 10(-4) to 1.40 x 10(-2)). Associations with all except +3228 C/T were replicated in the Twins UK cohort (Ps = 3.19 x 10(-9) to 6.00 x 10(-3)). In Chingford subjects, the 12 most common 8-SNP haplotypes (frequency 1.90%) explained 2.85% (p = 5.00 x 10(-2)) and in Twins UK subjects, the four most common 5-SNP haplotypes (frequency > 5.00%) explained 1.66% of the variance (p = 5.83 x 10(-7)). To investigate effects of -11391 G/A (rs17300539) and -11377 C/G (rs266729) on promoter activity, 1.2 kb of the ADIPOQ promoter region was cloned in a luciferase reporter plasmid, and the four haplotypes were transfected in differentiated 3T3-L1 adipocytes. No significant allelic effects on promoter activity were found.

Association of common JAK2 variants with body fat, insulin sensitivity and lipid profile.

The leptin signal is transduced via the JAK2-STAT3 (Janus kinase 2-signal transducer and activator of transcription-3) pathway at the leptin receptor. JAK2 also phosphorylates insulin receptor substrate, integral to insulin and leptin action and is required for optimum adenosine triphosphate-binding cassette transporter A1 (ABCA1)-dependent transport of lipids from cells to apolipoprotein A-1 (apoA-I). We hypothesized that common variation in the JAK2 gene may be associated with body fat, insulin sensitivity, and modulation of the serum lipid profile in the general population. Ten tagging single-nucleotide polymorphisms (SNPs) spanning the gene were genotyped in 2,760 white female twin subjects (mean age 47.3 +/- 12.6 years) from the St Thomas' UK Adult Twin Registry. Minor allele frequencies were between 0.170 and 0.464. The major allele of rs7849191 was associated with higher central fat (P = 0.030), percentage of central fat (P = 0.014) and waist circumference (P = 0.027) the major allele of rs3780378 with higher serum apoA (P = 0.026), total cholesterol (P = 0.014), low-density lipoprotein (LDL) cholesterol (P = 0.012) and lower triglyceride (P = 0.023). However, no associations were significant at a level which took account of multiple testing. Although JAK2 is a critical element in leptin and insulin signaling and has a role in cellular cholesterol transport, we failed to establish associations of common SNPs with relevant phenotypes in this human study.

Quantitative linkage analysis for pancreatic B-cell function and insulin resistance in a large twin cohort.

OBJECTIVE: Insulin resistance and disturbed glucose homeostasis are key characteristics of metabolic syndrome, diabetes, and cardiovascular disease. The recent nonlinear computer version of homeostasis model assessment (HOMA)2 provides an appropriate and convenient assessment of glucose metabolism, enabling gene-mapping studies in large population samples. RESEARCH DESIGN AND METHODS: Fasting insulin and glucose concentration were measured in 758 dizygous and 305 monozygous nondiabetic female pairs from the St. Thomas' U.K. adult twin registry (TwinsUK). Insulin resistance (IR) and pancreatic beta-cell function (BCF) were estimated from this data using the HOMA2 model. RESULTS: Genome-wide variance component linkage analysis using 2,231 genetic markers identified a highly significant quantitative trait locus for BCF on chromosome 10p15 (logarithm of odds [LOD] 6.2, P = 0.0001), a region recently shown to contain a functional variant for type 1 diabetes. Both BCF and IR suggested a pleiotropic effect on 17q25 (univariate LOD 3.2, P = 0.0012, and 2.38, P = 0.0087; bivariate LOD 2.66), and one additional region showed linkage for IR on chromosome 22q11 (LOD 3.2, P = 0.0016), providing replication and refining previous findings for diabetes and associated traits. CONCLUSIONS: To our best knowledge, this is the first genome-wide linkage screen for HOMA2 indexes in a large, healthy female sample. These results suggest that loci involved in control of normal glucose homeostasis among nondiabetic individuals might overlap with those involved in the development of diabetes. Linkage replications in independent studies and across populations provide information on important regions of common but potentially heterogeneous variability that can now be used for targeted positional candidate studies.

Relation of birth weight, body mass index, and change in size from birth to adulthood to insulin resistance in a female twin cohort.

CONTEXT AND OBJECTIVES: Because an adverse intrauterine environment is thought to induce insulin resistance, our objective was to investigate the relationships between birth weight, BMI, and change in body size over the life course and insulin resistance. SETTING, DESIGN, AND PARTICIPANTS: We conducted a cross-sectional study in a cohort of 1194 female twins aged 18-74 yr. The relationship between birth weight and insulin resistance was analyzed using a regression method allowing for a simultaneous estimation of within- and between-pair influences. The approach allows the influence of individual fetal nutrition on adult insulin resistance to be distinguished from effects that are mediated by confounding factors in the maternal environment. MAIN OUTCOME MEASURES: Insulin resistance was measured by the homeostasis model assessment. RESULTS: Individual level regression analyses showed no significant relationship between birth weight and insulin resistance. There was a significant positive relationship between insulin resistance and current body mass index (BMI) (a 26% increase in insulin resistance per sd increase in BMI; confidence interval, 22.6-29.5%). This significant relationship was accounted for in equal parts by individual-specific effects and by confounding factors in the shared environment of the twins. The relationship with birth weight became significant only after adjustment for BMI and was mediated only through between-pair differences. CONCLUSIONS: These results suggest that insulin resistance is influenced more by current body size than birth weight and that postnatal growth is potentially more important than fetal growth in the subsequent development of insulin resistance.

Intra-individual variation in creatinine and cystatin C.

BACKGROUND: Serum creatinine and cystatin C are measured in the assessment of glomerular filtration rate. Biological variation of an analyte is an important determinant of its usefulness. Intra-individual variation in serum creatinine and cystatin C in healthy subjects was determined in this study. METHODS: Weekly blood samples were taken from 10 healthy subjects and analysed for creatinine (kinetic Jaffe method) and for cystatin C (by nephelometry). RESULTS: Intra-individual variations for serum creatinine and cystatin C were 6.1% and 4.55%, respectively. Index of individuality values were similar for the analytes (0.35 for both). CONCLUSION: Intra-individual variations for serum creatinine and cystatin C were similar in healthy subjects.

Intrauterine, environmental, and genetic influences in the relationship between birth weight and lipids in a female twin cohort.

OBJECTIVE: To investigate the association between birth weight and lipid levels in a cohort of UK female twins. METHODS AND RESULTS: Birth weights and fasting blood lipid levels were available for 2900 women aged 18 to 80 years. Individual level regressions indicated that a 1-kg increase in birth weight was associated with a 0.08-mmol/L decrease in total cholesterol (95% confidence interval [CI], -0.12, -0.04) and a 0.06-mmol/L decrease in low-density lipoprotein (-0.10, -0.03). Using a regression model that includes both mean twin pair birth weight and individual twin's difference from the pair mean, we found that these significant relationships were between twin pairs only and not within pairs. We found no significant relationships for high-density lipoprotein. When monozygotic and dizygotic twins were analyzed separately we found similar effect sizes. Restricting the analysis to postmenopausal women we found stronger relationships between birth weight and lipid levels, which was attenuated after adjustment for body mass index (BMI). CONCLUSIONS: These novel results suggest that significant relationships between birth weight and lipids are mediated through shared influences on the maternal environment and do not support the hypothesis that fetal malnutrition is an important determinant of adult lipid levels. Adjustment for BMI also indicates that postnatal growth may be more important than prenatal growth.

Identification of QTLs for serum lipid levels in a female sib-pair cohort: a novel application to improve the power of two-locus linkage analysis.

Using a novel approach for a two-locus model that provides a greatly increased power to detect multiple quantitative trait loci (QTLs) in simulated data, we identified in a sample of 961 female sib-pairs, three genome-wide significant QTLs for apolipoprotein A1 on chromosomes 8p21.1-q13.1 (LOD score 3.71), 9q21.32-33.1 (LOD score 3.28) and 10p15.1-p13 (LOD score 5.51), two for lipoprotein (a) on chromosomes 6q25.2-q27 (LOD score 10.18) and 21q21.1-q21.3 (LOD score 4.57) and two for triglycerides on chromosomes 4q28.3-32.1 (LOD score 3.71) and 5q23.1-q32 (LOD score 3.60). The two-locus ordered-subset analysis has led to the confirmation of known and likely identification of novel regions linked to serum lipid levels that would have otherwise been missed and deserves wider application in linkage analyses of quantitative traits. Given the relative lack of power for the sample sizes commonly used in human genetics linkage studies, minor QTL effects often go undetected and those that are detected will be upwardly biased. We show through simulation that the discrepancy between the real and estimated QTL-effects is often likely to generate an unpredictable source of false-negative errors, using multi-locus models, reducing the power to detect multiple QTLs through oligogenic linkage analysis. The successful simultaneous modelling of the identified QTLs in a multi-locus context helps to eliminate false positives and increases the power to detect linkages, adding compelling evidence that they are likely to be reliable QTLs for these lipid traits.

Genetic and environmental influences on lipids, lipoproteins, and apolipoproteins: effects of menopause.

OBJECTIVE: Levels of lipids and (apo)lipoproteins are known to increase after menopause, but it is unknown whether the genetic and environmental variability alters or whether lipids and (apo)lipoproteins are influenced by different genes before and after menopause. METHODS AND RESULTS: We studied 453 monozygotic and 1280 dizygotic pairs of female white twins recruited from the St. Thomas' UK Adult Twin Registry and measured total cholesterol, low density lipoprotein (LDL), high density lipoprotein (HDL), triglycerides, lipoprotein(a) [Lp(a)], apolipoprotein A1 (apoA1), and apolipoprotein B (apoB). Variance components software was used to estimate genetic and environmental influences on serum lipid levels in premenopausal and postmenopausal women. Total variance was higher for triglycerides, HDL, and apoB after menopause. Postmenopausal women showed larger genetic variance for most lipids, apart from apoB and Lp(a). In premenopausal females, total cholesterol, LDL, HDL, apoA1, and apoB all showed an influence of the shared environment (22% to 34%), which, after menopause, decreased in HDL and completely disappeared in total cholesterol, LDL, and apoA1. Only for Lp(a), with a high heritability of 87%, did the same model fit premenopausal and postmenopausal women. Generally, there was no indication that different genes influence lipids before and after menopause. CONCLUSIONS: These findings imply that genetic studies of lipids can pool results from premenopausal and postmenopausal women and that family-based interventions, such as changes in diet, are more likely to succeed in younger women, in whom the environmental influences are greater.

Defective nitric oxide production and functional renal reserve in patients with type 2 diabetes who have microalbuminuria of African and Asian compared with white origin.

Diabetic nephropathy is a leading cause of end-stage renal failure. Its incidence is higher and is increasing in persons of Indo-Asian and African-Caribbean (African-Asian) compared with those of white origin. Nitric oxide deficiency is associated with progressive renal disease. It was hypothesized that differences in the capacity to increase glomerular filtration (functional renal reserve) would exist between these racial groups in relation to nitric oxide availability. Patients with type 2 diabetes of African-Asian (n = 9) and white (n = 9) origin with microalbuminuria were studied under euglycemic conditions. Glomerular filtration, renal plasma flow, and clearance of the stable metabolites of nitric oxide, nitrite, and nitrate were measured before and after a renal vasodilatory stimulus of a mixed amino acid intravenous infusion. There were no significant differences in age, duration of diabetes, and baseline glomerular filtration (57.1 [14.1] versus 55.8 [10.1] yr; P = 0.82, 14.5 [10.2] versus 9.1 [7.0] yr; P = 0.19 and 125.9 [30.9] versus 127.2 [44.6] ml/min per 1.73 m(2); P = 0.94) between the African-Asian and white groups. Functional renal reserve, change in renal plasma flow, and percentage change in nitrate and nitrite clearance was significantly higher in the white compared with the African-Asian group (21.9 [45.7] versus -2.5 [28.2] ml/min per 1.73 m(2); P = 0.043, 155.8 [205.9] versus -90.1 [146.0]; P = 0.03 ml/min per 1.73 m(2) and 26.7 [85.1] versus -44.7 [16.9] %; P = 0.013, respectively). The differences in functional reserve were not confounded after adjustment for diabetes duration (P = 0.034). The data suggest that these patients with type 2 diabetes of African and Asian origin lose functional renal reserve earlier in the evolution of nephropathy than whites. The differences appear to be due to defective nitric oxide production or bioavailability and might explain some of the propensity to develop end-stage renal disease.