Twin study
Twin studies are studies conducted on
Twins are a valuable source for observation because they allow the study of environmental influence and varying
Twins are also useful in showing the importance of the unique environment (specific to one twin or the other) when studying trait presentation. Changes in the unique environment can stem from an event or occurrence that has only affected one twin. This could range from a head injury or a birth defect that one twin has sustained while the other remains healthy.
The classical twin design compares the similarity of monozygotic (identical) and dizygotic (fraternal) twins. If identical twins are considerably more similar than fraternal twins (which is found for most traits), this implies that genes play an important role in these traits. By comparing many hundreds of families with twins, researchers can then understand more about the roles of genetic effects, shared environment, and unique environment in shaping behavior.
Modern twin studies have concluded that all studied traits are partly influenced by genetic differences, with some characteristics showing a stronger influence (e.g. height), others an intermediate level (e.g. personality traits) and some more complex heritabilities, with evidence for different genes affecting different aspects of the trait – as in the case of autism.[2]
History
Gustav III, King of Sweden was the first to commission a medical study using identical twins.[citation needed][original research?] Gustav's father, Adolph Frederick had been an opponent of stimulating drinks such as tea and coffee, signing the Misuse and Excesses Tea and Coffee Drinking Edict in 1757.[5] Both Gustav III and his father had read and been strongly influenced by a 1715 treatise from a French physician on the dangers of what would later be identified as caffeine in tea and coffee.[6] After assuming the throne in 1771 the king became strongly motivated to demonstrate to his subjects that coffee and tea had deleterious effects on human health. To this end he offered to commute the death sentences of a pair of twin murderers if they participated in a primitive clinical trial.
Both condemned men agreed and subsequently spent the rest of their lives in prison fulfilling the king's demands: that one twin drink three pots of coffee every day and the other three pots of tea. The tea drinking twin died first at the age of 83, long after Gustav III, who was assassinated in 1792. The age of the coffee-drinking twin at his death is unknown, as both doctors assigned by the king to monitor this study predeceased him. The ban on coffee and tea in Sweden was lifted in 1823.[7][8]
A more recent study is from
Thorndike incorrectly reasoned that his data supported for there being one, not two, twin types. This mistake was repeated by Ronald Fisher (1919), who argued
The preponderance of twins of like sex, does indeed become a new problem, because it has been formerly believed to be due to the proportion of identical twins. So far as I am aware, however, no attempt has been made to show that twins are sufficiently alike to be regarded as identical really exist in sufficient numbers to explain the proportion of twins of like sex.[12]
An early, and perhaps first, study understanding the distinction is from the German geneticist Hermann Werner Siemens in 1924.[13] Chief among Siemens' innovations was the polysymptomatic similarity diagnosis. This allowed him to account for the oversight that had stumped Fisher, and was a staple in twin research prior to the advent of molecular markers.
Wilhelm Weinberg and colleagues in 1910 used the identical-DZ distinction to calculate respective rates from the ratios of same- and opposite-sex twins in a maternity population. They partitioned co-variation amongst relatives into genetic and environmental elements, anticipating the later work of Fisher and Wright, including the effect of dominance on similarity of relatives, and beginning the first classic-twin studies.[14]
A study conducted by Darrick Antell and Eva Taczanowski found that "twins showing the greatest discrepancies in visible aging signs also had the greatest degree of discordance between personal lifestyle choices and habits", and concluded that "the genetic influences on aging may be highly overrated, with lifestyle choices exerting far more important effects on physical aging."[15]
Examples
Examples of prominent twin studies include the following:
- Maudsley Bipolar Twin Study
- Minnesota Twin Family Study
- Twins Early Development Study
- NASA Twins Study
Methods
The power of twin designs arises from the fact that twins may be either identical (
The basic logic of the twin study can be understood with very little mathematical knowledge beyond an understanding of the concepts of variance and thence derived correlation.
Classical twin method
Like all behavior genetic research, the classical twin study begins by assessing the variance of behavior (called a phenotype by geneticists) in a large group, and attempts to estimate how much of this is due to:
- genetic effects (heritability)
- shared environment – events that happen to both twins, affecting them in the same way
- unshared, or unique, or nonshared environment – events that occur to one twin but not the other, or events that affect either twin in a different way.
Typically these three components are called A (additive genetics) C (common environment) and E (unique environment); hence the acronym ACE. It is also possible to examine non-additive genetics effects (often denoted D for dominance (ADE model); see below for more complex twin designs).
The ACE model indicates what proportion of variance in a trait is heritable, versus the proportion due to a shared environment or unshared environment. Research is typically carried out using Structural equation modeling (SEM) programs such as OpenMx capable in principle of handling all sorts of complex pedigrees. However the core logic underlying such programs is the same as the one underlying the twin design described here.
Monozygotic (identical – MZ) twins raised in a family share 100% of their genes, and all of their shared environment. Any differences arising between them in these circumstances are random (i.e. due to environmental effects unique to each twin). The correlation between identical twins provides an estimate of A + C. Dizygotic (DZ) twins also share C, but share, on average only 50% of their genes: so the correlation between fraternal twins is a direct estimate of ½A+C. If we denote with r the correlation, we can define rmz and rdz as the correlations of a trait among identical and fraternal twins, respectively. For any particular trait, then:
- rmz = A + C
- rdz = ½A + C
Stated again, the difference between these two sums then allows us to solve for A and C (and as a consequence, for E). As the difference between the identical and fraternal correlations is due entirely to a halving of the genetic similarity, the additive genetic effect A is twice the difference between the identical and fraternal correlations:
- A = 2 (rmz − rdz)
given the estimate for A, the one for C can be derived, for instance, from the first equation:
- C = rmz − A
Finally, since the trait correlation among identical twins reflects the full contribution of A and C, the residual variation E can be estimated by subtracting this correlation from 1
- E = 1 − rmz.
To summarize therefore, the additive genetic factor A is twice the difference between MZ and DZ twin correlations (this is known as Falconer's formula), C is the MZ twin correlation minus this estimate of A, and the random (unique) factor E is (1 - rmz), i.e. MZ twins differ due to unique environments only (Jinks & Fulker, 1970; Plomin, DeFries, McClearn, & McGuffin, 2001).
Underestimation of the genetical effect
The effect of genes may be much higher than what is predicted by naïve twin models, because they assume that non-identical twins are just half closer to identical twins in genetical similarity than random people. Due to assortative mating, they are even more similar, as parents are genetically more similar to each other than random people. Taking this into account (NTFD method), Wolfram and Morris showed that the number of years in education was just 26% affected by the shared environment, not 43%.[19]
Modern modeling
Beginning in the 1970s, research transitioned to
An example structural model (for the heritability of height among Danish males)[20] is shown:
Model A on the left shows the raw variance in height. This is useful as it preserves the absolute effects of genes and environments, and expresses these in natural units, such as mm of height change. Sometimes it is helpful to standardize the parameters, so each is expressed as percentage of total variance. Because we have decomposed variance into A, C, and E, the total variance is simply A + C + E. We can then scale each of the single parameters as a proportion of this total, i.e., Standardised–A = A/(A + C + E). Heritability is the standardised genetic effect.
Model comparison
A principal benefit of modeling is the ability to explicitly compare models: Rather than simply returning a value for each component, the modeler can compute
Multi-group and multivariate modeling
Multivariate modeling can give answers to questions about the genetic relationship between variables that appear independent. For instance: do IQ and long-term memory share genes? Do they share environmental causes? Additional benefits include the ability to deal with interval, threshold, and continuous data, retaining full information from data with missing values, integrating the latent modeling with measured variables, be they measured environments, or, now, measured molecular genetic markers such as SNPs. In addition, models avoid constraint problems in the crude correlation method: all parameters will lie, as they should, between 0–1 (standardized).
Multivariate, and multiple-time wave studies, with measured environment and repeated measures of potentially causal behaviours are now the norm. Examples of these models include extended twin designs,[21][22] simplex models,[23] and growth-curve models.[24]
SEM programs such as OpenMx[25] and other applications suited to constraints and multiple groups have made the new techniques accessible to reasonably skilled users.
Modeling the environment: MZ discordant designs
As MZ twins share both their genes and their family-level environmental factors, any differences between MZ twins reflect E: the unique environment. Researchers can use this information to understand the environment in powerful ways, allowing
An example of a positive MZ discordant effect is shown below on the left. The twin who scores higher on trait 1 also scores higher on trait 2. This is compatible with a "dose" of trait 1 causing an increase in trait 2. Of course, trait 2 might also be affecting trait 1. Disentangling these two possibilities requires a different design (see below for an example). A null result is incompatible with a causal hypothesis.
Take for instance the case of an observed link between depression and exercise (See Figure above on right). People who are depressed also reporting doing little physical activity. One might hypothesise that this is a
Longitudinal discordance designs
As may be seen in the next Figure, this design can be extended to multiple measurements, with consequent increase in the kinds of information that one can learn. This is called a cross-lagged model (multiple traits measured over more than one time).[27]
In the longitudinal discordance model, differences between identical twins can be used to take account of relationships among differences across traits at time one (path A), and then examine the distinct hypotheses that increments in trait1 drive subsequent change in that trait in the future (paths B and E), or, importantly, in other traits (paths C & D). In the example, the hypothesis that the observed correlation where depressed persons often also exercise less than average is causal, can be tested. If exercise is protective against depression, then path D should be significant, with a twin who exercises more showing less depression as a consequence.
Assumptions
It can be seen from the modeling above, the main assumption of the twin study is that of equal family environments, also known as the equal environments assumption.[28][29][30] A special ability to test this assumption occurs where parents believe their twins to be non-identical when in fact they are genetically identical. Studies of a range of psychological traits indicate that these children remain as concordant as MZ twins raised by parents who treated them as identical.[31]
Molecular genetic methods of heritability estimation have tended to produce lower estimates than classical twin studies due to modern SNP arrays not capturing the influence of certain types of variants (e.g., rare variants or repeat polymorphsisms), though some have suggested it is because twin studies overestimate heritability.[32] A 2016 study determined that the assumption that the prenatal environment of twins was equal was largely tenable.[33] Researchers continue to debate whether or not the equal environment assumption is valid.[34][35][36][37][38]
Measured similarity: A direct test of assumptions in twin designs
A particularly powerful technique for testing the twin method was reported by Visscher et al.[39] Instead of using twins, this group took advantage of the fact that while siblings on average share 50% of their genes, the actual gene-sharing for individual sibling pairs varies around this value, essentially creating a continuum of genetic similarity or "twinness" within families. Estimates of heritability based on direct estimates of gene sharing confirm those from the twin method, providing support for the assumptions of the method.
Sex differences
Genetic factors, including both gene expression and the range of gene × environment interactions, may differ between the sexes. Fraternal opposite sex twin pairs are invaluable in explicating these effects.
In an extreme case, a gene may only be expressed in one sex (qualitative sex limitation).[clarification needed] More commonly, the effects of particular alleles may depend on the sex of the individual. A gene might cause a change of 100 g in weight in males, but perhaps 150 g in females – a quantitative gene effect.
Environments may impact on the ability of genes to express themselves and may do this via sex differences. For instance, genes affecting voting behavior would have no effect in females if females are excluded from the vote. More generally, the logic of sex-difference testing can extend to any defined sub-group of individuals. In cases such as these, the correlation for same and opposite sex DZ twins will differ, betraying the effect of the sex difference.
For this reason, it is normal to distinguish three types of fraternal twins. A standard analytic workflow would involve testing for sex-limitation by fitting models to five groups, identical male, identical female, fraternal male, fraternal female, and fraternal opposite sex. Twin modeling thus goes beyond correlation to test causal models involving potential causal variables, such as sex.
Gene × environment interactions
Gene effects may often be dependent on the environment. Such interactions are known as G×E interactions, in which the effects of a gene allele differ across different environments. Simple examples would include situations where a gene multiplies the effect of an environment: perhaps adding 1 inch to height in high nutrient environments, but only half an inch to height in low-nutrient environments. This is seen in different slopes of response to an environment for different genotypes.
Often researchers are interested in changes in
A second effect is G × E correlation, in which certain alleles tend to accompany certain environments. If a gene causes a parent to enjoy reading, then children inheriting this allele are likely to be raised in households with books due to GE correlation: one or both of their parents has the allele and therefore will accumulate a book collection and pass on the book-reading allele. Such effects can be tested by measuring the purported environmental correlate (in this case books in the home) directly.
Often the role of environment seems maximal very early in life, and decreases rapidly after compulsory education begins. This is observed for instance in reading[40] as well as intelligence.[41] This is an example of a G*Age effect and allows an examination of both GE correlations due to parental environments (these are broken up with time), and of G*E correlations caused by individuals actively seeking certain environments.[42]
Norms of reaction
Studies in plants or in
As in other fields such as
While the twin study tells us only how genes and families affect behavior within the observed range of environments, and with the caveat that often genes and environments will covary, this is a considerable advance over the alternative, which is no knowledge of the different roles of genes and environment whatsoever.
Extended twin designs and more complex genetic models
The basic or classical twin-design contains only identical and fraternal twins raised in their biological family. This represents only a sub-set of the possible genetic and environmental relationships. It is fair to say, therefore, that the heritability estimates from twin designs represent a first step in understanding the genetics of behavior.
The variance partitioning of the twin study into additive genetic, shared, and unshared environment is a first approximation to a complete analysis taking into account
An initial limitation of the twin design is that it does not afford an opportunity to consider both Shared Environment and Non-additive genetic effects simultaneously. This limit can be addressed by including additional siblings to the design.
A second limitation is that gene–environment correlation is not detectable as a distinct effect unless it is added to the model. Addressing this limit requires incorporating adoption models, or children-of-twins designs, to assess family influences uncorrelated with shared genetic effects.
Continuous variables and ordinal variables
While concordance studies compare traits either present or absent in each twin, correlational studies compare the agreement in continuously varying traits across twins.
Criticism
The twin method has been subject to criticism from statistical genetics, statistics and psychology, with some researchers, such as Burt & Simons (2014), arguing that conclusions reached via this method are ambiguous or meaningless.[51] Core elements of these criticisms and their rejoinders are listed below.
Criticisms of fundamental assumptions
Critics of twin studies argue that they are based on false or questionable assumptions, including that monozygotic twins share 100% of their genes
Criticisms of statistical methods
Responses to statistical critiques
Before computers, statisticians used methods that were computationally tractable, at the cost of known limitations. Since the 1980s these approximate statistical methods have been discarded. Modern twin methods based on structural equation modeling are not subject to the limitations and heritability estimates such as those noted above are mathematically impossible.[61] Critically, the newer methods allow for explicit testing of the role of different pathways and incorporation and testing of complex effects.[47]
Sampling: Twins as representative members of the population
Results of twin studies cannot be automatically generalized beyond the population they come from. It is therefore important to understand the particular sample studied, and the nature of twins themselves. Twins are not a
For example: Dizygotic (DZ) twin births are affected by many factors. Some women frequently produce more than one
Response to representativeness of twins
However, twins differ very little from non-twin siblings. Measured studies on the personality and intelligence of twins suggest that they have scores on these traits very similar to those of non-twins (for instance Deary et al. 2006).
Separated twin pairs as representative of other twins
Separated twin pairs, identical or fraternal, are generally separated by adoption. This makes their families of origin non-representative of typical twin families in that they give up their children for adoption. The families they are adopted to are also non-representative of typical twin families in that they are all approved for adoption by children's protection authorities and that a disproportionally large fraction of them have no biological children. Those who volunteer to studies are not even representative of separated twins in general since not all separated twins agree to be part of twin studies.[69][70]
Detection problems
There can be some issues of undetected behaviors in the case of behaviors that many people keep secret presently or in their earlier lives. They may not be as willing to reveal behaviors that are discriminated against or stigmatized. If environment played no role in the actual behavior, skewed detection would still make it look like it played a role. For environment to appear to have no role in such cases, there would have to be either a counterproductivity of intolerance in the sense of intolerance causing the behavior it is bigoted against, or a flaw in the study that makes the results scientifically useless. Even if environment does play a role, the numbers would still be skewed.[71][72][73]
Terminology
Pairwise concordance
For a group of twins, pairwise concordance is defined as C/(C+D), where C is the number of concordant pairs and D is the number of discordant pairs.
For example, a group of 10 twins have been pre-selected to have one affected member (of the pair). During the course of the study four other previously non-affected members become affected, giving a pairwise concordance of 4/(4+6) or 4/10 or 40%.
Probandwise concordance
For a group of twins in which at least one member of each pair is affected, probandwise concordance is a measure of the proportion of twins who have the illness who have an affected twin and can be calculated with the formula of 2C/(2C+D), in which C is the number of concordant pairs and D is the number of discordant pairs.
For example, consider a group of 10 twins that have been pre-selected to have one affected member. During the course of the study, four other previously non-affected members become affected, giving a probandwise concordance of 8/(8+6) or 8/14 or 57%.
See also
- Behavioral genetics
- "Burt Affair"
- Gene-environment interaction
- Gene-environment correlation
- Genome-wide complex trait analysis
- Heritability
- Heritability of IQ
- Human nature
- Identical Strangers: A Memoir of Twins Separated and Reunited
- Kaiser Wilhelm Institute of Anthropology, Human Heredity, and Eugenics
- Michigan State University Twin Registry
- Minnesota Twin Family Study
- Nature versus nurture
- Otmar Freiherr von Verschuer
- The Parent Trap
- Quantitative genetics
- Differential susceptibility
- Three Identical Strangers
- Twin registry
- TwinsUk
References
- ^ Plomin, R.; DeFries, J. C.; Knopik, V. S.; Neiderhiser, J. M. (Ed.). (2014). Behavioral Genetics (6th ed.). New York, NY: Worth Publishers
- S2CID 205349969.
- ISBN 9780801840906.
- ^ Cicero, De Divinatione, (On Divination), ii. 42
- PMID 28348964.
- ^ "Linné on line – Coffee – rat poison or miracle medicine?". www2.linnaeus.uu.se. Retrieved 2022-10-04.
- PMID 28348964.
- S2CID 244415520.
- S2CID 22666939.
- JSTOR 2011451. Retrieved 2019-03-11.
- ^ Thorndike, Edward Lee (1905). Measurements of Twins. Science Press.
- PMID 17245935.
- OCLC 18362377.
- PMID 10388804.
- PMID 10597816.
- ISBN 978-0-471-94174-3.
- ISBN 978-0-7923-1874-3.
- ^ S2CID 2028886.
- ^ Tobias Wolfram & Damien Morris (July 2023). "Conventional twin studies overestimate the environmental differences between families relevant to educational attainment". npj Science of Learning. 8 (24).
- S2CID 2235255.
- PMID 20013306.
- PMID 15989749.[permanent dead link]
- PMID 15607015. Archived from the original(PDF) on 2010-06-24. Retrieved 2010-11-05.
- PMID 11035490.
- PMID 23258944.
- PMID 18678794.
- PMID 19899929.
- PMID 16238870.
- S2CID 30732913.
- ^ Winerman, Lea (2004-04-01). "Behavioral Genetics--A second look at twin studies". Monitor on Psychology. Retrieved 2017-08-23.
- S2CID 30324607.
- PMID 24267761.
...estimates of cumulative genetic influence using molecular-level data have tended to be substantially lower than the corresponding estimates from twin studies.
- PMID 26410687..
- S2CID 2083213.
- ^ Fosse, Roar, Jay Joseph, and Ken Richardson. "A critical assessment of the equal-environment assumption of the twin method for schizophrenia." Frontiers in psychiatry 6 (2015): 62.
- .
- ^ Joseph, Jay. The trouble with twin studies: A reassessment of twin research in the social and behavioral sciences. Routledge, 2014.
- PMID 24267761.
- PMID 16565746.
- S2CID 144069119. Archived from the original(PDF) on 2016-03-04. Retrieved 2015-08-28.
- PMID 16721405.
- PMID 21807642.
- S2CID 23266944.
- PMID 6732417.
- ISBN 978-0-14-022605-8.
- PMID 17821364.
- ^ a b c M. C. Neale and H. H. Maes. (1996). Methodology for genetics studies of twins and families. Journal.
- .
- PMID 20644632.
- PMID 17766918.
- ^ .
- PMID 23865113.
- S2CID 7229563.
- PMID 27906529.
- S2CID 143865688.
- PMID 24267761.
- JSTOR 2826060.
- S2CID 37292855.
- ^ Schönemann, Peter H. (1995). Totems of the IQ Myth: General Ability (g) and its Heritabilities (h2, HR). 1995 Meetings of the American Association for the Advancement of Sciences.
- S2CID 34798197.
- ^ M. C. Neale, S. M. Boker, G. Xie and H. H. Maes. (2002). Mx: Statistical Modelling. Journal.
- S2CID 30109683.
- OCLC 12445470.
- PMID 2924931.
- S2CID 919014.
- S2CID 27421288.
- ISBN 978-0-7100-9888-7.
- ^ Capron, Christiane; Vetta, Adrian R.; Duyme, Michel; Vetta, Atam (1999). "Misconceptions of biometrical IQists". Cahiers de Psychologie Cognitive/Current Psychology of Cognition. 18 (2): 115–160.
- ^ Fatal Flaws in the Twin Study Paradigm: A Reply to Hatemi and Verhulst, Doron Shultziner 2013
- ^ Twin Studies of Political Behavior: Untenable Assumptions?, Jon Beckwith and Corey A. Morris 2008
- ^ Critical Analysis: A Comparison of Critical Thinking Changes in Psychology and Philosophy Classes, Teaching of Psychology 2014 41: 28
- ^ Association for Psychological Science: Why Science Is Not Necessarily Self-Correcting, John P. A. Ioannidis 2012
- ^ How Black African and White British Women Perceive Depression and Help-Seeking: a Pilot Vignette Study, International Journal of Social Psychiatry March 2010
Further reading
- Free courseware, textbook, software, and example scripts for twin research
- Jang, K.L.; McCrae, R.R.; Angleitner, A. Riemann; Livesley, W.J. (1998). PMID 9654759.
- R. Plomin, J. C. DeFries, V. S. Knopik and J. M. Neiderhiser. (2012). Behavioral Genetics. Worth Publishers. London
- Nancy L. Segal (2005) Indivisible by Two: Lives of Extraordinary Twins. New York, Harvard University Press.
- Segal, Nancy L. (2012). Born Together—Reared Apart. Cambridge, MA: Harvard University Press. ISBN 978-0-674-05546-9.
- Bryan Caplan (June 20, 2012). "O Brother, Who Art Thou?". Wall Street Journal.
- Am J Med Genet C Semin Med Genet. 2009 May 15;151C(2):136-41. Not really identical: epigenetic differences in monozygotic twins and implications for twin studies in psychiatry. Haque FN, Gottesman II, Wong AH.
Critical accounts
- S2CID 36598943. Archived from the original(PDF) on 2008-02-28.
- Schönemann, Peter; Schönemann, Roberta D. (1994). "Environmental versus genetic models for Osborne's personality data on identical and fraternal twins" (PDF). CPC. 13 (2): 141–167. Archived from the original (PDF) on 2012-07-22. Retrieved 2013-07-06.
- Kamin, L. J. (1974). The Science and Politics of I.Q. Potomac, MD: Lawrence Erlbaum Associates.
- Kempthorne, O (1997). "Heritability: uses and abuses". Genetica. 99 (2–3): 109–112. S2CID 23266944.
- Joseph, J. (2003). The Gene Illusion: Genetic Research in Psychiatry and Psychology Under the Microscope. PCCS Books.
- This book has been critically reviewed for the American Psychological Association. Hanson, D. R. (2005). 'The Gene Illusion Confusion: A review of The Gene Illusion: Genetic Research in Psychiatry and Psychology Under the Microscope by Jay Joseph' [Electronic Version]. PsycCritiques, 50, e14.
- Capron, Christiane; Vetta, Adrian R.; Duyme, Michel; Vetta, Atam (1999). "Misconceptions of biometrical IQists". Cahiers de Psychologie Cognitive/Current Psychology of Cognition. 18 (2): 115–160.
- Horwitz, AV; Videon, TM; Schmitz, MF; Davis, D (Jun 2003). "Rethinking twins and environments: possible social sources for assumed genetic influences in twin research". J Health Soc Behav. 44 (2): 111–129. PMID 12866384.
- And in reply to this article see:
- Freese, J; Powell, B (Jun 2003). "Tilting at Twindmills: rethinking sociological responses to behavioral genetics". J Health Soc Behav. 44 (2): 130–135. PMID 12866385.
- Freese, J; Powell, B (Jun 2003). "Tilting at Twindmills: rethinking sociological responses to behavioral genetics". J Health Soc Behav. 44 (2): 130–135.
External links
Academic bodies
Several academic bodies exist to support behavior genetic research, including the Behavior Genetics Association, the International Society for Twin Studies, and the International Behavioural and Neural Genetics Society. Behavior genetic work also features prominently in several more general societies, for instance the International Society of Psychiatric Genetics.
Journals
Prominent specialist journals in the field include