Additive,Epistatic, and Environmental Effects Through the Lens of Expression Variability QTL in a Twin Cohort

The expression of a gene can vary across individuals in the general population, as well as between monozygotic twins. This variable expression is assumed to be due to the influence of both genetic and nongenetic factors. Yet little evidence supporting this assumption has been obtained from empirical data. In this study, we used expression data from a large twin cohort to investigate the influences of genetic and nongenetic factors on variable gene expression. We focused on a set of expression variability QTL (evQTL)—i.e., genetic loci associated with the variance, as opposed to the mean, of gene expression. We identified evQTL for 99, 56, and 79 genes in lymphoblastoid cell lines, skin, and fat, respectively. The differences in gene expression, measured by the relative mean difference (RMD), tended to be larger between pairs of dizygotic (DZ) twins than between pairs of monozygotic (MZ) twins, showing that genetic background influenced the expression variability. Furthermore, a more profound RMD was observed between pairs of MZ twins whose genotypes were associated with greater expression variability than the RMD found between pairs of MZ twins whose genotypes were associated with smaller expression variability. This suggests that nongenetic (e.g., environmental) factors contribute to the variable expression. Lastly, we demonstrated that the formation of evQTL is likely due to partial linkages between eQTL SNPs that are additively associated with the mean of gene expression; in most cases, no epistatic effect is involved. Our findings have implications for understanding divergent sources of gene expression variability.     

Steroid metabolism in primates. XVIII. Return to higher levels of 17-ketosteroid excretion in urine after ending a long-term treatment with mestranol/chlormadinone acetate in the baboon]

6 weeks after terminating a long-time application of the ovulation inhibitor Ovosiston (mestranol/chlormadinone acetate) in female baboons, excretion of 17-ketosteroids in urine is still decreased. 6 months after ceasing the preparation, urinary 17-ketosteroid excretion resembles that of the control group.     

Does higher concordance in monozygotic twins than in dizygotic twins suggest a genetic component?

It is widely regarded that twins can be used as a natural experiment to subject hypotheses to empirical testing regarding the contributions of genetic factors to phenotypic variability in human traits, especially behavioral traits. In genetic epidemiology, a higher concordance rate in monozygotic (MZ) twins than in dizygotic (DZ) twins is often taken as prima facie evidence for a genetic component. While twins studies have been used to estimate the contributions of genetic factors to phenotypic variability in human traits, the corresponding methodology that allows the estimation entails several crucial assumptions. The most critical is that MZ and DZ twins are equally similar environmentally. Although MZ twins are genetically more similar than DZ twins, they are often environmentally more similar. This paper demonstrates that, even in the complete absence of any genetic factor and of any biases, the greater environmental similarity alone in MZ twins can result in higher concordance rate in MZ twins than in DZ twins. This is especially true when there are multiple environmental factors, which may have multiple exposure levels and/or interact strongly, although each of them may be of low risk. This may serve as a sobering antidote to the uncritical reliance on twin studies without examining the validity of the underlying assumptions.     

Color perception in twins

The classical twin method was used to examine the genotype--phenotype relationship in color vision. Suprathreshold color differences were assessed by 5 pairs of monozygotic (MZ) and 3 pairs of dizygotic (DZ) twins. The control group included 3 unrelated normal trichromats, a non-twin sibling pair, and a previously diagnosed deuteranomal. Concordance rates were calculated by Spearman's correlation coefficients (rs) and Procrustean distances (gl) between the reconstructed color spaces for each related pair of observers. For 4 pairs of the MZ twins, the rs values were comparable to intraindividual variability in the control normal trichromat; they were significantly higher (0.94-0.97) than those for the DZ twins and siblings (0.72-0.82). The gl values for the MZ twins (0.008-0.029) were lower than for the DZ twins (0.073-0.079) and siblings (0.053). The high concordance between each pair of the MZ twins suggests that their shared photopigment genome constrains a contribution of possible individual variations in nongenetic factors to variability of their color spaces. Lower concordance rates in the DZ twins and siblings can be attributed to differences in the inherited arrays of photopigment genes. Contributions to intrapair variation in color spaces of twins from cognitive factors such as perceptual-cognitive color categorization and decision-process variability are discussed.     

Utilizing Twins as Controls for Non-Twin Case-Materials in Genome Wide Association Studies

Twin registries around the globe have collected DNA samples from large numbers of monozygotic and dizygotic twins. The twin sample collections are frequently used as controls in disease-specific studies together with non-twins. This approach is unbiased under the hypothesis that twins and singletons are comparable in terms of allele frequencies; i.e. there are no genetic variants associated with being a twin per se. To test this hypothesis we performed a genome-wide association study comparing the allele frequency of 572,352 single nucleotide polymorphisms (SNPs) in 1,413 monozygotic (MZ) and 5,451 dizygotic (DZ) twins with 3,720 healthy singletons. Twins and singletons have been genotyped using the same platform. SNPs showing association with being a twin at P-value < 1 × 10^-5 were selected for replication analysis in 1,492 twins (463 MZ and 1,029 DZ) and 1,880 singletons from Finland. No SNPs reached genome-wide significance (P-value < 5 × 10^-8) in the main analysis combining MZ and DZ twins. In a secondary analysis including only DZ twins two SNPs (rs2033541 close to ADAMTSL1 and rs4149283 close to ABCA1) were genome-wide significant after meta-analysis with the Finnish population. The estimated proportion of variance on the liability scale explained by all SNPs was 0.08 (P-value=0.003) when MZ and DZ were considered together and smaller for MZ (0.06, P-value=0.10) compared to DZ (0.09, P-value=0.003) when analyzed separately. In conclusion, twins and singletons can be used in genetic studies together with general population samples without introducing large bias. Further research is needed to explore genetic variances associated with DZ twinning.     

THE CLINICAL SIGNIFICANCE OF URINARY 17-KETOSTEROID ASSAYS

The urinary 17-ketosteroids are a group of compounds derived from complex steroids produced by the adrenal cortex, testis, or ovary. The method of determining the amount excreted has been simplified so that it is available for routine diagnostic purposes. Usually the amount is increased in diseases in which there is hyperfunction of the adrenal cortex due to tumor or hyperplasia, and decreased in lesions that impair the function of the adrenal cortex.Other conditions such as myxedema, eunuchism, gout, and arthritis may alter the excretion of the 17-ketosteroids. Low levels are also found in the young and in the aged. Case histories are presented to illustrate the findings in the following diseases: Tumors of the adrenal cortex with (a) masculinization, (b) Cushing''s syndrome with virilism and, (c) hirsutism; as well as in gigantism with acromegaly, in gout, eunuchism, Addison''s disease, myxedema, and severe panhypopituitarism.     

Causes of blood methylomic variation for middle-aged women measured by the HumanMethylation450 array

To address the limitations in current classic twin/family research on the genetic and/or environmental causes of human methylomic variation, we measured blood DNA methylation for 479 women (mean age 56 years) including 66 monozygotic (MZ), 66 dizygotic (DZ) twin pairs and 215 sisters of twins, and 11 random technical duplicates using the HumanMethylation450 array. For each methylation site, we estimated the correlation for pairs of duplicates, MZ twins, DZ twins, and siblings, fitted variance component models by assuming the variation is explained by genetic factors, by shared and individual environmental factors, and by independent measurement error, and assessed the best fitting model. We found that the average (standard deviation) correlations for duplicate, MZ, DZ, and sibling pairs were 0.10 (0.35), 0.07 (0.21), -0.01 (0.14) and -0.04 (0.07). At the genome-wide significance level of 10^?7, 93.3% of sites had no familial correlation, and 5.6%, 0.1%, and 0.2% of sites were correlated for MZ, DZ, and sibling pairs. For 86.4%, 6.9%, and 7.1% of sites, the best fitting model included measurement error only, a genetic component, and at least one environmental component. For the 13.6% of sites influenced by genetic and/or environmental factors, the average proportion of variance explained by environmental factors was greater than that explained by genetic factors (0.41 vs. 0.37, P value <10^?15). Our results are consistent with, for middle-aged woman, blood methylomic variation measured by the HumanMethylation450 array being largely explained by measurement error, and more influenced by environmental factors than by genetic factors.     

Gender difference in the Oatp1-mediated tubular reabsorption of estradiol 17beta-D-glucuronide in rats

The gender difference in the urinary excretion of estradiol-17beta-glucuronide (E(2)-17betaG) was examined in rats. The urinary clearance of E(2)-17betaG was >250 times lower in male than in female rats. No such major gender difference was observed in its biliary excretion or metabolism in kidney homogenate. Both plasma protein binding and inulin clearance were comparable in male and female rats, suggesting that this gender difference cannot be explained by glomerular filtration. The urinary clearance with respect to the plasma unbound E(2)-17betaG in male rats was <1% of the glomerular filtration rate, indicating its potential reabsorption by the kidney, and this increased to a level comparable with that found in female rats when dibromosulfophthalein was coinfused. A marked increase in E(2)-17betaG urinary excretion was also observed in male rats that had undergone orchidectomy. Testosterone injections given to female rats reduced the urinary excretion to a level comparable with that of control male rats. The concomitant change in the expression of the gene product for organic anion-transporting polypeptide Oatp1, of which E(2)-17betaG is a typical substrate, was found in the kidney membrane fractions after these treatments. These results suggest that urinary E(2)-17betaG excretion is subject to hormonal regulation and that the large gender difference can be explained by regulation in Oatp1-mediated reabsorption.     

Genetic and environmental influences on bilateral dental asymmetry in Northwest Indian twins

The role of genetic and environmental factors on dental asymmetry (in maximum crown dimensions) was examined using 58 pairs of twins (23 MZ and 35 DZ) from Chandigarh, India. The t'-test for equality of means by zygosity showed only one variable significantly different among 56: this is ascribable to Type 1 error. Heterogeneity of MZ-DZ total variance was observed in 42.9% of traits of the two types (fluctuating and directional) of bilateral asymmetry. In general, MZ twins showed higher total variance than DZ pairs. MZ twins also showed stronger environmental covariance for a majority of the traits. Dental asymmetry measures thus yielded consistently low genetic variance ratios and indicated predominantly complex environmental determinism. Since fluctuating asymmetry is widely believed to be an environmental stress indicator, this data set allows confirmation of methods for detecting unequal environmental influences on the zygosities which bias estimates of genetic variance and heritability.     

Role of genetic and environmental influences on heart rate variability in middle-aged men

Our aim was to estimate causal relationships of genetic factors and different specific environmental factors in determination of the level of cardiac autonomic modulation, i.e., heart rate variability (HRV), in healthy male twins and male twins with chronic diseases. The subjects were 208 monozygotic (MZ, 104 healthy) and 296 dizygotic (DZ, 173 healthy) male twins. A structured interview was used to obtain data on lifetime exposures of occupational loading, regularly performed leisure-time sport activities, coffee consumption, smoking history, and chronic diseases from 12 yr of age through the present. A 5-min ECG at supine rest was recorded for the HRV analyses. In univariate statistical analyses based on genetic models with additive genetic, dominance genetic, and unique environmental effects, genetic effects accounted for 31-57% of HRV variance. In multivariate statistical analysis, body mass index, percent body fat, coffee consumption, smoking, medication, and chronic diseases were associated with different HRV variables, accounting for 1-11% of their variance. Occupational physical loading and leisure-time sport activities did not account for variation in any HRV variable. However, in the subgroup analysis of healthy and diseased twins, occupational loading explained 4% of the variability in heart periods. Otherwise, the interaction between health status and genetic effects was significant for only two HRV variables. In conclusion, genetic factors accounted for a major portion of the interindividual differences in HRV, with no remarkable effect of health status. No single behavioral determinant appeared to have a major influence on HRV. The effects of medication and diseases may mask the minimal effect of occupational loading on HRV.     

Covariability factors affecting urinary vanilmandelic acid excretion and height in various human populations

The urinary excretion of vanilmandelic acid (VMA), a final product of catecholamine metabolism, has been studied in relation to height in various human groups. A positive correlation between these two variables has been demonstrated. The VMA-height relationship between groups is of a different nature than the relationship between individuals of each group. The influence of socio-professional and nutritional factors has also been taken into account. The observations are discussed in the light of Schreider's work on height, mental characteristics and socio-professional status. A hypothesis is proposed to explain how these variables and the catecholamine metabolism might share some variability factors.     

Kynurenine metabolism in rats: some hormonal factors affecting enzyme activities.

Six male subjects (19–23 years old) underwent a 7-day control period with respect to diet, temperature (22C), and sleep (7.5 hrs), followed by a 2-day exposure to 15C and a 2-day recovery period (22C). Urine collections were made every 8 hours commencing at 2300 hours; MHPG and VMA were assayed using gas-liquid chromatography. During the control period a diurnal rhythmicity was demonstrated for MHPG and VMA with maxima at 0700–1500 hours. The mean excretory rates for MHPG and VMA were 0.71 ± 0.04 μg and 2.6 ± 0.2 μg per milligram creatinine (± S.E.), respectively. Cold exposure abolished the rhythms for MHPG and VMA and caused an 18% increase in MHPG excretion. In contrast, VMA excretion was not altered. Significant correlations were obtained with MHPG excretion and both urinary cortisol and rectal temperature. The data suggest that MHPG excretion may be indicative of changes in norephinephrine metabolism in the central nervous system, although alterations in peripheral degradative pathways cannot be ruled out. Careful interpretation of changes in MHPG excretion in clinical studies is emphasized due to the relative ease of altering MHPG metabolism.     

Effect of allyl isothiocyanate on plasma and urinary concentrations of some biochemical entities in the rat.

Allyl isothiocyanate (AITC) is a constituent of several plants of the family Cruciferae that are commonly used as food. This study investigated the effect of feeding AITC to male Sprague-Dawley rats on their plasma glucose and uric acid levels as well as on the urinary concentrations of glucose, 17-ketosteroids (17-KS), creatinine, and uric acid. Other test compounds included were thyroxine (T4) and thiouracil (TU). AITC caused a highly significant (P smaller than or equal to 0.01) depression in the plasma glucose and uric acid levels compared with the control. TU caused a significant depression only of the plasma glucose. T4, on the other hand, significantly increased the levels of both glucose and uric acid. The AITC-treated rats voided twice as much urine as the controls or those receiving TU or injected with T4. The 24-h excretion of glucose, uric acid, and creatinine was significantly (P smaller than or equal to 0.01) higher in animals fed AITC than in those consuming the control diet, while the excretion of 17-KS was significantly lower. Results on an equal urine volume basis showed that differences in the excretion of glucose and creatinine were related to differences in the urine volume. TU significantly depressed excretion of all the compounds but glucose. The effect of T4 on the excretion of 17-KS and uric acid resembled that of AITC and TU, thus showing that these compounds depressed the androgenic function of the animal.     

Cooperation and Competition as a Function of Zygosity in 7- to 9-Year-Old Twins

A study of 31 monozygotic (MZ) and 24 dizygotic (DZ) Singapore twin pairs (aged 7.6 to 9.7 years) is reported. MZ and DZ twins differed in cooperative and competitive behaviors measured by within-pair reactions to payoff structures designed to elicit simultaneous cooperation (SCO) or reciprocal altruism (RA) on a marble-pull task. Reciprocation entailed turn-taking in rewards. DZ twins showed greater competition than MZ twins when competing for marbles in the RA condition, as predicted. Unexpectedly, however, MZ twins were markedly more competitive than DZ twins under the SCO condition, when success on the task always rewarded both twins. The competition arose over which end of the apparatus was to be used on a given trial. This interaction between condition and zygosity could not be related to differences in parents’ reported treatment of their children or zygosity beliefs, nor did it appear to be related to differences in factors such as time spent together and shared activities. An explanation in terms of uncertain or unstable dominance in MZ twinships is suggested.     

Parent ratings of temperament in twins: explaining the 'too low' DZ correlations.

Twin studies of child temperament using objective measures consistently suggest moderate heritability for most dimensions. However, parent rating measures produce unusual patterns of results. Intraclass correlations for identical (MZ) twins are typically high, whereas fraternal (DZ) twin intraclass correlations are much lower than would be predicted from an additive genetic model. The 'too low' DZ correlations can be explained by parent-rating biases that either exaggerate the differences between DZ twins (contrast effects) or that inflate the similarity of MZ twins (assimilation effects), or by the presence of non-additive genetic variance. To evaluate the three possible explanations, we used model-fitting procedures applied to parent-rating data averaged across 14, 20, 24, and 36 months of age in a sample of 196 twin pairs participating in the MacArthur Longitudinal Twin Study. The data were best described by a model that included contrast effects. Implications for non-twin research are discussed.     

The effects of chorion type on variation in IQ in the NCPP twin population.

The 7-year IQ scores (WISC) of 116 white and 143 black nonmalformed twins of known zygosity and placental type were ascertained from the NINCDS Collaborative Perinatal Project (NCPP). The type of chorion and zygosity had no significant effect on the mean IQ or among-pair variation. In white monozygotic twins, however, analysis of variance revealed a significantly greater within-pair mean square for dichorionic twins than monochorionic twins. On the other hand, the white dichorionic monozygotic (MZ) and dizygotic (DZ) within-pair mean squares were quite similar. These findings were not evident in blacks for either of the within-pair comparisons. In addition, estimates of genetic variance were dependent upon MZ chorion type in both races. These data suggest to us that (1) in white twin pairs dichorionic placentas are of greater influence than the similarity or dissimilarity of genomes with regard to intrapair IQ development, and (2) failure to consider chorion type may introduce a serious bias in the interpretation of genetic variance estimates of IQ variability.     

Liquid chromatography–electrospray tandem mass spectrometry of acidic monoamine metabolites

A new method based on liquid chromatography–tandem mass spectrometry has been developed for the determination of monoamine metabolites, i.e., homovanillic acid (HVA), vanilmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC) and 5-hydroxyindoleacetic acid (5-HIAA) in human urine. Analytes were separated on a C₁₆ amide (5 cm, 5 μm) column and ionized by negative ion electrospray. Operating in the selected-reaction monitoring mode, linearity was established over three-orders of magnitude and limits of detection were in the range 30–70 μg/l. Precision calculated as RSD was within 0.8–5.2% for all intra- and inter-day determinations. The method was applied to the quantitative analysis of monoamine metabolites in 700 urine samples from occupationally (adults) and environmentally (both children and adults) exposed people living in areas with different soil contamination from lead. The urinary excretion of monoamine metabolites was significantly higher (P<0.001) in the subgroup of children living in polluted areas as compared to the control group (HVA, 6.03 vs. 4.57 mg/g creatinine; VMA, 5.33 vs. 4.37 mg/g creatinine; 5-HIAA 3.24 vs. 2.45 mg/g creatinine). In adults belonging to both groups of subjects occupationally and environmentally exposed, no differences were detected in the urinary concentration of monoamine metabolites. However, adults showed lower values of HVA (2.57 mg/g creatinine), VMA (2.17 mg/g creatinine) and 5-HIAA (2.09 mg/g creatinine) as compared to children groups.     

Quantitative hematological characteristics in MZ and DZ twins

The level of several biochemical hematological characteristics was determined--using colorimetric methods--in a group of 111 twins, aged 16-22 years. Significant variance differences between MZ and DZ twin pairs were observed in metabolites and serum enzyme levels. The intrapair differences of the variance of ions and protein levels were comparable in MZ and DZ twins. The validity of quantitative determination of heterability of biochemical characteristics is discussed.     

Inheritance of finger pattern types in MZ and DZ twins

Digital patterns of a sample on twins were analyzed to estimate the resemblance between monozygotic (MZ) and dizygotic (DZ) twins and to evaluate the mode of inheritance by the use of maximum likelihood based variance decomposition analysis. MZ twin resemblance of finger pattern types appears to be more pronounced than in DZ twins, which suggests the presence of genetic factors in the forming of fingertip patterns. The most parsimonious model shows twin resemblance in count of all three basic finger patterns on 10 fingers. It has significant dominant genetic variance component across all fingers. In the general model, the dominant genetic variance component proportion is similar for all fingertips (about 60%) and the sibling environmental variance is significantly nonzero, but the proportion between additive and dominant variance components was different. Application of genetic model fitting technique of segregation analyses clearly shows mode of inheritance. A dominant genetic variance component or a specific genetic system modifies the phenotypic expression of the fingertip patterns. The present study provided evidence of strong genetic component in finger pattern types and seems more informative compared to the earlier traditional method of correlation analysis.