Genetic linkage analysis for complex diseases offers a major challenge to geneticists. In these complex diseases multiple genetic loci are responsible for the disease and they may vary in the size of their contribution; the effect of any single one of them is likely to be small. In many situations, like in extensive twin registries, trait values have been recorded for a large number of individuals, and preliminary studies have revealed summary measures for those traits, like mean, variance and components of variance, including heritability. Given the small effect size, a random sample of twins will require a prohibitively large sample size. It is well known that selective sampling is far more efficient in terms of genotyping effort. In this paper we derive easy expressions for the information contributed by sib pairs for the detection of linkage to a quantitative trait locus (QTL). We consider random samples as well as samples of sib pairs selected on the basis of their trait values. These expressions can be rapidly computed and do not involve simulation. We extend our results for quantitative traits to dichotomous traits using the concept of a liability threshold model. We present tables with required sample sizes for height, insulin levels and migraine, three of the traits studied in the GenomEUtwin project.

The relative importance of genetic influences (heritability) on five general textural quality characteristics of the human iris was assessed using sex and age limitation models. Colour photographs of irises were available from 100 monozygotic twin pairs, 99 dizygotic twin pairs, and 99 unrelated randomly paired age-matched German subjects. Comparative scales were constructed and two judges who were blind to zygosity independently rated five characteristic of the subjects' left iris. Inter-rater reliabilities were larger than .90 for all five scales. The heritabilities for the five iris characteristics ranged from .51–.90. No sex-specific genetic factors were found for the iris characteristics. Age-group differences in heritability were found for one of the five iris characteristics — “distinction of white dot rings”. Heritability was greater for the older cohort (90%) than the younger (73%). The iris characteristics that showed the next highest additive-genetic effect were “contractional furrows” (78%) and “frequency of crypts” in the main stroma leaf (66%).

We report on a woman who gave birth to dichorionic diamniotic twins with a birthweight discordancy of 30%. Her placenta exhibited characteristic features. The placental villous tissues were completely separated, but there was marked vascular communication between the two placentas. The lighter twin received less blood than the heavier twin via these vascular communications. Although the details are unknown, this abnormal placental structure may have caused the weight discordancy in this case. Clinicians must pay attention to placental vascular communication as one possible cause of twin discordancy in not only monochorionic but also dichorionic twins.

Twin studies typically indicate shared environmental influence for cognitive abilities, especially in early childhood. However, across studies, DZ twin correlations tend to be greater than non-twin sibling correlations, suggesting that twin estimates of shared environment are to some extent specific to twins. We tested this hypothesis in a sample of more than 1800 MZ and 1800 same-sex DZ pairs from the Twins Early Development Study (TEDS), a study of twins born in England and Wales in 1994 and 1995. For this analysis, we obtained comparable data from more than 130 same-sex younger siblings of the twins. Twins and their younger siblings were assessed for language, cognitive abilities and behavior problems by their parents at 2 and 3 years of age. For language and cognitive measures at both 2 and 3 years, but not for behavior problems, estimates of shared environment were more than twice as large for twins as compared to non-twin siblings. We conclude that about half of twin study estimates of shared environment for cognitive abilities in early childhood are specific to twins. Although many possibilities exist for explaining the special shared environment effect for twins, we suggest that cognitive-relevant experiences that are not shared by siblings are shared by twins because they are exactly the same age.

Database infrastructure has become a critical component for competitive life sciences research and discovery. The explosion of data requires that the data are properly loaded, accessed, managed, queried, analyzed, and shared with others. The key purpose of the population-based twin cohorts housed at different institutions in Europe is to gather an extremely large quantity of information from their twin populations, and share it. Longitudinal research over a long period of time, hopefully generations, demands completely new methods and systems to handle the gathering of information and storing. These cohorts bring to the fore problems concerning the need for a standardization of research data and a computer and storage strategy. In the following we describe the preliminary strategy being implemented in the Database Core of GenomEUtwin.

Studies in Sweden and Australia have examined the relationship between number of CAG repeats in the androgen receptor gene and psychological traits — three Masculinity-Femininity (M-F) measures in Australia, and the Karolinska Scales of Personality (KSP) in Sweden. The present study derived M-F scales from the KSP items, and scales corresponding to several KSP scales from the items of the inventories used in Australia, to permit cross-validation of the Australian results in the Swedish sample, and vice versa. The derivation of scales was facilitated by the fact that items from both inventories had been used with a large twin sample in the US. Correlation of the derived scales with androgen receptor gene CAG-repeat scores for women in the Australian and Swedish samples failed to provide clear evidence of replication of either set of original correlations in the other sample, although there were a few hints of consistency. It was concluded that if the number of CAG repeats on this gene is related to psychological traits at all, the relationship is a weak one.

For zygosity diagnosis in the absence of genotypic data, or in the recruitment phase of a twin study where only single twins from same-sex pairs are being screened, or to provide a test for sample duplication leading to the false identification of a dizygotic pair as monozygotic, the appropriate analysis of respondents' answers to questions about zygosity is critical. Using data from a young adult Australian twin cohort (N = 2094 complete pairs and 519 singleton twins from same-sex pairs with complete responses to all zygosity items), we show that application of latent class analysis (LCA), fitting a 2-class model, yields results that show good concordance with traditional methods of zygosity diagnosis, but with certain important advantages. These include the ability, in many cases, to assign zygosity with specified probability on the basis of responses of a single informant (advantageous when one zygosity type is being oversampled); and the ability to quantify the probability of misassignment of zygosity, allowing prioritization of cases for genotyping as well as identification of cases of probable laboratory error. Out of 242 twins (from 121 like-sex pairs) where genotypic data were available for zygosity confirmation, only a single case was identified of incorrect zygosity assignment by the latent class algorithm. Zygosity assignment for that single case was identified by the LCA as uncertain (probability of being a monozygotic twin only 76%), and the co-twin's responses clearly identified the pair as dizygotic (probability of being dizygotic 100%). In the absence of genotypic data, or as a safeguard against sample duplication, application of LCA for zygosity assignment or confirmation is strongly recommended.

We investigated the etiology of six problem behaviors that might facilitate an understanding of behavioral pathways to substance use and abuse in adolescents. These behavioral measures, classified as Conduct Problems, Hyperactivity, School Problems, Low Self-esteem, Neuroticism, and Social Withdrawal were the result of a previously reported (Siewert et al., 2003) modification of the Drug Use Screening Inventory (DUSI; Tarter, 1990; Tarter & Hegedus, 1991). We developed these measures as interpretable components of risk for substance use and abuse in a community based sample of 633 twin pairs, who were under the legal drinking age of 21 (mean age = 15.0 years). Using multivariate analyses, model comparisons indicated that these six behavioral measures could be thought of as two heritable, and genetically distinct, dimensions of problem behavior. Two closely competing models resulted from our analyses. The best fitting model hypothesized a general genetic factor loading on all 6 behavioral measures with a second genetic factor loading on only the three internalizing behavioral measures with loadings of 0.25–0.59 and 0.26–0.44, respectively. A second model, which fit the data almost as well, hypothesized one genetic factor loading only on the externalizing behavioral measures, and a second genetic factor loading only on the internalizing behavioral measures, with a correlation between the two latent factors of 0.75. Because our analyses show that there are two genetically distinct factors influencing these six problem behaviors, we anticipate that there may be different patterns of relationship of these factors to risk for substance use, abuse, and dependence.

Monoamniotic twinning is a rare event with an incidence of 1% of all monozygotic twins and associated with a high fetal morbidity and mortality. Confident early diagnosis is possible, but optimal management is not yet established. This article presents the experience of a single centre in managing all monoamniotic twins diagnosed during 1994–2000. Seven pairs of monoamniotic twins were identified for analysis. All were managed in accord with a unit protocol that involved early diagnosis, serial ultrasound examination and elective early delivery. In four cases, the detection of monoamnionicity was made during a first trimester nuchal scan. Discordance for structural abnormality was found in three cases where the co-twin was normal. Cord entanglement was detected antenatally in four cases. Two pairs of twins died before 20 weeks. One of these had early onset twin–twin transfusion syndrome. In five cases, the pregnancy continued beyond 20 weeks. A live birth rate of 90% and intact survival of 70% were achieved in this group. We believe that ultrasound is reliable in diagnosing monoamniotic twins and the detection of cord entanglement. Timing of elective delivery is a balance between the risks of preterm birth at a specific gestational age in an individual centre compared with the unquantifiable risks of fetal death if an expectant policy were pursued. The decision to deliver and at which gestational age should combine input from the parents, neonatologist, fetal medicine consultant and the obstetrician.

We compared twins to their gender-matched singleton classmates in peer-assessed behavioral adjustment. Our samples include 1874 11- to 12-year-old Finnish twins (687 monozygotic, MZ; 610 same-sex dizygotic, SSDZ; 577 opposite-sex dizygotic, OSDZ) and their 23,200 non-twin classmates. Data were collected using a 30-item Multidimensional Peer Nomination Inventory containing three factors and their subscales. We found twin-singleton differences: classmates rated twin girls and boys higher than gender-matched singletons in Adaptive Behaviors (constructive, compliant, and socially active behavior), and those effects were particularly evident among OSDZ twins for assessments of social interaction, popularity, and leadership. We found no evidence that individual twins differ from singletons in Externalizing (hyperactivity- impulsivity, inattention, aggression) or Internalizing Problem Behaviors (depressive symptoms, social anxiety). Nor did we find systematic differences between MZ and SSDZ twins. Among both twins and singletons, boys exceeded girls in Externalizing, and girls exceeded boys in Internalizing Problem Behaviors. Results suggest that a twinship forms a positive developmental environment for socioemotional behavior, particularly among OSDZ twins.

It has been suggested that twinning may influence handedness through the effects of birth order, intra-uterine crowding and mirror imaging. The influence of these effects on handedness (for writing and throwing) was examined in 3657 Monozygotic (MZ) and 3762 Dizygotic (DZ) twin pairs (born 1893–1992). Maximum likelihood analyses revealed no effects of birth order on the incidence of left-handedness. Twins were no more likely to be left-handed than their singleton siblings (n = 1757), and there were no differences between the DZ co-twin and siblingtwin covariances, suggesting that neither intra-uterine crowding nor the experience of being a twin affects handedness. There was no evidence of mirror imaging; the co-twin correlations of monochorionic and dichorionic MZ twins did not differ. Univariate genetic analyses revealed common environmental factors to be the most parsimonious explanation of familial aggregation for the writing-hand measure, while additive genetic influences provided a better interpretation of the throwing hand data.

The relationship between regular smoking behavior and the smoking behavior of parents, siblings and friends was investigated using data from the Netherlands Twin Register. Cross-sectional analyses of data of 3906 twins showed significant associations between smoking behavior of the participant and smoking behavior of co-twin, additional brothers, parents of the same sex as the participant and friends. Those variables, together with age, explained 47% of the variance in smoking behavior. Longitudinal analyses of data from 2397 twins, who, in 1993, reported never to have smoked (regularly), showed that uptake of regular smoking two years later was predicted by having a smoking co-twin, smoking same-sex siblings, smoking mother and smoking friends. Males are, in contrast to females, at a later age still vulnerable to taking up regular smoking. The variables explained 21% of the variance. Sport participation, alcohol use, boredom susceptibility and neuroticism significantly added to the predictive value of this model. Including those additional factors increased the explained variance to 30%, and subsequently adding experimental smoking behavior further increased the explained variance to almost 50%. In summary, having smoking family members and friends, as well as lifestyle and personality factors are important predictors for the uptake of regular smoking. However, the experimental smoking behavior of the participant is equally important.

Dizygotic twinning in humans is influenced by genetic factors suggesting inherited variation affects follicle development and predisposes to double ovulations. In a previous study, we conducted a detailed examination of follicle development and variation in hormone concentrations during the menstrual cycle in mothers of DZ twins (MODZT) compared with an age-matched control group of mothers of singletons. We did not detect differences in FSH concentrations between mothers of twins and mothers of singletons. Serum inhibin concentrations were measured by a radioimmunoassay that did not distinguish between dimeric inhibin A and B forms and free inhibin α subunit. We therefore analyzed the samples from this study with specific assays to determine whether concentrations of inhibin A and B were different between MODZT and controls and therefore contribute to the twinning phenotype. There were no significant differences between MODZT with single ovulations and control women in inhibin A and B concentrations during the cycle, including the critical period for the selection of the dominant follicle. These data suggest that the genetic cause of twinning is not associated with changes in FSH concentrations or recognised feedback mechanisms regulating FSH release.

Multiallelic short tandem repeat polymorphisms, or microsatellites, are useful markers in genome wide scans to identify chromosomal regions containing genes underlying disease loci. The biallelic single nucleotide polymorphism (SNP) can be used to fine map previously identified large candidate regions or to test functional candidate genes by association analysis. In the GenomEUtwin project the population based impact of susceptibility genes for six multifactorial traits will be studied. A genome wide panel of informative human microsatellite markers will be analyzed by fluorescent capillary electrophoresis in well characterized twin and population samples. Contrary to microsatellites, selection of the most informative panels of SNPs is hampered by imperfect data on the allele frequencies and population distribution of SNPs markers in the databases. Therefore, selection of SNPs requires a substantial amount of bioinformatics, and, the SNPs need to be validated experimentally in the relevant populations prior to genotyping large sample sets. In the GenomEUtwin project, large scale genotyping of SNPs will be performed using the SNPstreamUHT and MassARRAY genotyping systems that are based on the primer extension reaction principle combined with fluorescent and mass spectrometric detection, respectively. Production of the genotyping data will be a joint effort by GenomEUtwin partners at the University of Helsinki, the National Public Health Institute in Helsinki, Finland and Uppsala University, Sweden. All genotyping data will be stored in a common database established specifically for the GenomEUtwin project, from where it can be accessed by the twin research centres that provided the samples for genotyping.

Questionnaire-based zygosity assessment in twins has generally been found to be valid. In this report we evaluate sex- and age-differences in the validity of such questionnairebased classification when using the four questions that have been the basis of zygosity assessment in The Danish Twin Registry for half a century. Three hundred and forty-two male and 531 female twin pairs were zygosity diagnosed using genetic markers and the results compared with the original questionnaire based classification. We found significant differences in the accuracy of questionnaire based zygosity diagnosis when stratifying the data for sex as well as age: males and monozygotic having the highest misclassification. However, even in the group with the highest misclassification rate the frequency was less than 8%. The overall misclassification rate was only 4%, with a clear tendency towards a higher proportion of misclassified monozygotic than dizygotic twins. The results demonstrate that questionnaire based zygosity diagnosis can still be regarded as a valid and valuable classification method for most purposes.

Amajor component of variation in body height is due to genetic differences, but environmental factors have a substantial contributory effect. In this study we aimed to analyse whether the genetic architecture of body height varies between affluent western societies. We analysed twin data from eight countries comprising 30,111 complete twin pairs by using the univariate genetic model of the Mx statistical package. Body height and zygosity were self-reported in seven populations and measured directly in one population. We found that there was substantial variation in mean body height between countries; body height was least in Italy (177 cm in men and 163 cm in women) and greatest in the Netherlands (184 cm and 171 cm, respectively). In men there was no corresponding variation in heritability of body height, heritability estimates ranging from 0.87 to 0.93 in populations under an additive genes/unique environment (AE) model. Among women the heritability estimates were generally lower than among men with greater variation between countries, ranging from 0.68 to 0.84 when an additive genes/shared environment/unique environment (ACE) model was used. In four populations where an AE model fit equally well or better, heritability ranged from 0.89 to 0.93. This difference between the sexes was mainly due to the effect of the shared environmental component of variance, which appears to be more important among women than among men in our study populations. Our results indicate that, in general, there are only minor differences in the genetic architecture of height between affluent Caucasian populations, especially among men.

Increasing numbers of women wish to breastfeed their multiple birth children. Breastfeeding of preterm and fullterm multiple birth infants is complex and demanding for the families and presents distinct challenges for health professionals. Families require sustained assistance from health care providers who are encouraging, knowledgeable, skilled, and committed to the breastfeeding of multiple birth children. Seven breastfeeding rights of multiple birth families are presented for the continuum of pregnancy to early childhood and are in accordance with the Declaration of Rights and Statement of Needs of Twins and Higher Order Multiples (Council of Multiple Birth Organizations of the International Society for Twin Studies, 1995). Guidelines for each of the rights have been developed to assist health professionals provide “best practices” in community and hospital settings. The guidelines are based on the existing body of breastfeeding of multiples' research, empirical findings, and consultations with parents and care providers with experience and/or expertise in breastfeeding multiples. The rights and guidelines suggest direction for providing assistance, implementing programs and services, conducting research, and evaluating the effectiveness of multiples-specific breastfeeding care during the prenatal, infancy, and toddlerhood periods.

Research on unusually long birth intervals between twin and triplet deliveries is reviewed. A recent multicentre study reported gestational and survival data for 35 multiple birth sets from 12 centers, and evaluated conservative treatment strategies for mothers delivering one twin fetus. Next, findings from three recent twin studies are reviewed: effects of close inter-twin communication on longevity; case report of dizygous twins with a single chorion; and a case of discordance for hemihypertrophy in an MZ female twin pair. The article concludes with summaries of three interesting and informative anecdotal accounts of twins.

Twins are prone to developmental delay due to prematurity and low birthweight. However it is unknown if twinning is an independent risk factor for developmental delay. The objective of this study was to compare the attainment of a set of gross motor milestones in a cohort of twins and singletons in The Gambia. Eighty-four pairs of twins and 72 singletons were enrolled at birth and followed up until 18 months of age. The mean age at achieving milestones was higher in twins for each development outcome and the difference between twins and singletons was significant after adjustment for confounders for maintaining head, sitting without support and walking. In twins, we found a highly significant correlation within pairs for most milestones. When monozygotic and dizygotic twins were compared, a significant heritability was observed for crawling, sitting, standing and walking, with over 90% of population variance observed due to genetic factors rather than environmental factors. There was little evidence for a genetic contribution towards very early milestones. In conclusion, our data suggest that twinning is an independent risk factor for developmental delay in early life in The Gambia, and that genetic factors contribute strongly to certain motor development outcomes.