Carpenter syndrome

Alcohol use during pregnancy is a significant public health problem. Although most women who drink before pregnancy substantially reduce their consumption or completely stop drinking once they become pregnant, approximately 14 to 22.5 percent of women report drinking some alcohol during pregnancy (Bearer 2001).

The costs of prenatal alcohol use are high. Risky drinking (defined as more than seven standard drinks (1) per week or five or more standard drinks on a drinking day [CDC 2002]) during pregnancy is a primary risk factor for fetal alcohol syndrome (FAS), the most common preventable cause of mental retardation. (2) Prenatal alcohol exposure also can result in fetal alcohol spectrum disorders (FASD), in which the affected children do not show the classical FAS pattern but nonetheless exhibit mental, developmental, behavioral, and social deficits as well as other birth defects. Some evidence indicates that even low-risk drinking (defined as fewer than seven standard drinks per week or three or fewer standard drinks per drinking day [NIAAA 1995]) during pregnancy can cause adverse fetal effects, but how this damage occurs is not fully understood (Bearer 2001). An estimated 1 percent of all live-born infants show some prenatal alcohol-related damage, contributing to societal costs estimated at between $75 million and $9.7 billion per year (May and Gossage 2001).

Because of alcohol's adverse effects on the fetus, all women should be counseled to refrain from drinking during pregnancy. Tests that could identify women who continue to drink while pregnant--and could detect the effects of alcohol exposure on the developing fetus or newborn--would be invaluable for several reasons. Identifying these women would facilitate interventions that could help them stop using alcohol during pregnancy, thereby minimizing alcohol's effects on fetal brain development. Even discovering prenatal alcohol use later in pregnancy or soon after birth would be important because it would identify infants who are at risk for alcohol-associated birth defects and could make it possible to monitor them for potential problems, facilitate a more stable living environment, and provide special services if needed (Bearer et al. 1999; Stoler and Holmes 1999, 2004). In particular, identifying at-risk children before age 6 reduces the likelihood of secondary problems associated with FAS and FASD, such as mental health problems, school failure, delinquency, inappropriate sexual behavior, and alcohol and other drug problems (Streissguth et al. 1996). In addition, interventions that help new mothers reduce problem drinking could enhance their ability to care for their children and reduce the risk of alcohol problems during their subsequent pregnancies (Russell et al. 1996).

Developing effective biomarkers of prenatal alcohol use also may promote better scientific understanding of alcohol effects that occur with different patterns of maternal alcohol use during pregnancy.

Maternal Self-Report

Currently no laboratory test can identify and quantify prenatal alcohol use that takes place over a protracted period. Because alcohol itself and the main product of its metabolism, acetaldehyde, break down rapidly in the blood, they cannot be used to distinguish between a single drinking episode and chronic, intermittent alcohol use. Testing blood, breath, or urine is useful only for assessing very recent alcohol exposure. Because biological markers currently in use may not be effective in screening for risky alcohol use occurring over the longer term, such as during pregnancy, clinicians most commonly use brief screening measures that rely on maternal self-reports to assess drinking patterns (Chang 2001; Russell et al. 1994, 1996; Savage et al. 2002).

Major disadvantages of such screening measures are that it often is difficult for people to recall the amount and frequency of their alcohol intake, and the stigma and fear of punishment (e.g., incarceration or involuntary commitment) associated with drinking alcohol during pregnancy can make women reluctant to reveal prenatal alcohol use, especially if they drink heavily (Bearer et al. 2003; Chan et al. 2003).

Some screening instruments attempt to circumvent pregnant women's reluctance to disclose prenatal alcohol use by including questions that assess prenatal alcohol use indirectly--for example, by asking women to report the number of drinks they can consume before passing out or falling asleep (Russell et al. 1994, 1996). Some research indicates that such screening instruments can effectively flag heavy drinking. However, they do not provide a long-term, objective measure of the full range of prenatal alcohol use. Supplementing these measures with a biological marker for prenatal alcohol use would allow earlier identification and intervention for exposed infants and would make it easier to recognize women who are at risk for drinking during their next pregnancy (Bearer et al. 2001, 2003).

Biological Samples for Detecting Drinking During Pregnancy

Traditionally, samples of neonatal or maternal urine and blood have been used to determine prenatal alcohol use (Halmesmaki et al. 1992; Wisniewski et al. 1983). However, the biomarkers that have been measured in these samples mostly reflect alcohol exposure only in the 2 to 3 days before delivery. Neonatal urine is difficult to collect, and blood collection for a neonate is an invasive procedure. Other samples that could be used after delivery to assess biomarkers of prenatal alcohol consumption include amniotic fluid, cord blood, neonatal hair, placenta, breast milk, the infant's first fecal material (meconium), and the cheese-like material that covers the skin of a fetus (vernix). (The table summarizes information on potential maternal, fetal, and newborn samples.)

The usefulness of different fluid and tissue samples depends not only on their biochemical suitability but also on how readily they can be collected and how large the resulting sample is (Bearer 2001). For example, although amniotic fluid, cord blood, and placenta afford large samples, the window of opportunity for obtaining them is narrow. Likewise, analyses of maternal or neonatal hair can indicate the timing of prenatal alcohol use, but samples may not be available, either because a parent does not consent or, in the case of neonatal hair, because the newborn has none to give.

Meconium, most frequently used to detect prenatal exposure to drugs such as cocaine, holds particular promise for assessing prenatal alcohol use. Several studies have used meconium to measure concentrations of fatty acid ethyl esters (FAEEs), one promising biomarker of prenatal alcohol use (Bearer et al. 1999, 2003; Chan et al. 2003; Niemela et al. 1991). (See "Fatty Acid Ethyl Esters," below.) Meconium is available beginning shortly after birth, and large samples can be collected directly from the infant's diaper. Moreover, meconium begins to form as early as the 13th week of pregnancy and accumulates thereafter. Thus, using meconium sampled after delivery, it may be possible to obtain a detailed history of prenatal alcohol use over a longer period than is the case for biological samples such as neonatal urine or cord blood (Bearer et al. 1999, 2003; Chan et al. 2003).

A disadvantage of meconium as a matrix for detecting biomarkers of prenatal alcohol use is that it is present only for the first 2 or 3 days after birth. In contrast, neonatal hair, if present, can reflect maternal prenatal alcohol use for up to 2 or 3 months after birth; and the mother's hair may be obtained throughout pregnancy to determine prenatal alcohol use during the pregnancy.

Promising Biomarkers

Although some biomarkers may directly detect the effect of alcohol on the fetus or newborn (e.g., Bearer et al. 2000, 2001; Stoler et al. 1998), more progress has been made in developing biomarkers for maternal alcohol use during pregnancy. Research examining such biomarkers typically assesses their ability to correctly identify pregnant women who use alcohol (sensitivity) as well as pregnant women who do not (specificity).

Blood/Urine Marker Batteries. In an effort to develop a definitive biomarker for prenatal alcohol use, investigators have explored several biochemical changes associated with alcohol use, which can be detected by blood and/or urine tests:

* Long-term alcohol use causes the liver to release an enzyme called gamma-glutamyltransferase (GGT) into the bloodstream.

* Blood concentrations of variants of the protein transferrin (carbohydrate-deficient transferrin, or CDT) increase after alcohol consumption.

* An increase in the average size of red blood cells (i.e., mean corpuscular volume, or MCV) can signal excessive alcohol consumption.

* High levels of chemicals known as dolichols are found in the urine of alcoholics and are measurable in the urine of newborns whose mothers drank heavily during pregnancy (Halmesmaki et al. 1992; Wisniewski et al. 1983).

* When alcohol breaks down, its major metabolite, acetaldehyde, binds with hemoglobin in the blood, forming compounds known as hemoglobin--acetaldehyde adducts. These compounds remain in the blood longer than free acetaldehyde, making them a potential marker for long-term alcohol use (Niemela et al. 1991).

Although these biological changes are useful indicators of risky drinking, no single marker is diagnostically sensitive and specific enough to be considered a definitive biomarker for prenatal alcohol use (Bearer 2001; Stoler et al. 1998). Several investigations have used panels of two or more blood markers in an effort to achieve greater sensitivity and specificity (Halmesmaki et al. 1992; Sarkola et al. 2000; Stoler et al. 1998). For example, in a study of 529 women, Stoler and colleagues (1998) found that a combination of CDT, MCV, GGT, and hemoglobin-acetaldehyde adducts was more strongly correlated with prenatal alcohol use (assessed by evaluating newborns for facial features indicating the effects of prenatal alcohol exposure) than was any single biomarker or the maternal self-report of alcohol use during pregnancy.

Another study of pregnant women who drank eight or more drinks per week found that hemoglobin-acetaldehyde adducts and CDT were not associated with the reported level of drinking (Sarkola et al. 2000). However, MCV and GGT were significantly higher in women drinking at least eight drinks per week compared with those drinking fewer than eight drinks per week. The specificity and sensitivity of these markers were not reported. Other studies have similarly suggested that using multiple biomarkers for prenatal alcohol use may be effective, but the sensitivity and specificity of combined blood markers, and thus their clinical usefulness, remain unclear.

Fatty Acid Ethyl Esters (FAEEs). Fatty acid ethyl esters are metabolic products that result from the interaction between alcohol and fatty acids. FAEEs have been detected in the cord blood, meconium, and hair of infants; in other organs in adults; and in the placenta in mice (Bearer et al. 1999). Compared with alcohol and acetaldehyde, FAEEs have a prolonged half-life and show promise as biomarkers for prenatal alcohol use (Bearer et al. 1999, 2003; Chan et al. 2003; Klein et al. 1999). In a recent study, the presence of one FAEE (ethyl oleate) in meconium was strongly associated with self-reported drinking during the second and third trimesters of pregnancy, with very high sensitivity and specificity (see the figure) (Bearer et al. 2003). Further, the study found that FAEE concentration in meconium was more strongly related to the mother's self-reported alcohol consumption per occasion than to the overall average she consumed per week. This finding may prove especially useful, as recent research has suggested that the number of drinks per occasion may be the best indicator of risk for alcohol-related impairment (CDC 2002).

[GRAPHIC OMITTED]

Recently, a study found elevated FAEE concentrations in the hair of a newborn whose mother was alcoholic, as well as in the mother's hair (Klein et al. 2002). Although preliminary, this finding indicates that both maternal and neonatal hair may be a useful matrix for examining FAEE concentrations.

The correlation between FAEEs in meconium and prenatal alcohol use is not perfect (Bearer et al. 2003; Chan et al. 2003). There probably are several reasons for this: (1) FAEEs may accumulate unevenly in meconium over time, so that samples do not appear to reflect reported drinking; (2) genetically determined variations in alcohol metabolism may influence the synthesis of FAEEs; and (3) illness or the use of some medications and food additives may affect FAEE concentrations. In order to set appropriate cutoff concentrations of FAEEs for analyses, a better understanding of these issues is needed.

Proteomics

A number of proteins are known to be affected, either directly or indirectly, by alcohol. The rapidly advancing field of proteomics (3) offers promise for developing sophisticated biomarkers that can detect very subtle biological changes associated specifically with alcohol use (Anni and Israel 2002), or can distinguish between currently drinking women who will continue to drink during pregnancy and those who will stop (Neuhold et al. 2004).

So far, few investigators have focused attention on proteomic analyses designed to establish potential biomarkers for prenatal alcohol exposure. Robinson and colleagues (1995), in a preliminary study, searched for serum protein variations associated with FAS. These researchers studied 12 children diagnosed with FAS (indicating that prenatal alcohol exposure had to have occurred) and 8 control subjects. Their analysis found eight proteins whose concentrations differed significantly between the two groups of children. No single protein distinguished children with FAS from children in the control group, but analyses revealed clusters of proteins that collectively distinguished between the two groups. This study demonstrates the power of looking at patterns of response to alcohol, and the approach it describes can be used as a strategy not only to identify children with prenatal alcohol damage but also women who, because of their biological response to drinking, may be at risk for drinking during their pregnancies.

Robinson and colleagues' results, although not conclusive, illustrate the exciting potential of proteomic analysis to uncover novel biomarkers of both alcohol use and risk for drinking during pregnancy. The development of such proteomic biomarkers would be a major step toward primary prevention of the disorders associated with prenatal alcohol use.

Conclusion

Currently, no laboratory test can definitively detect and quantify prenatal alcohol use. Developing effective biomarkers is an important step toward identifying at-risk pregnancies, preventing alcohol-related birth defects, and diagnosing and intervening with infants who may be at risk for later problems because of prenatal alcohol exposure. Several lines of research are promising, but much research is still needed to validate potentially useful biomarkers and identify further areas of possibility.

(1) The National Institute on Alcohol Abuse and Alcoholism (NIAAA) defines a standard drink as 11-14 g of alcohol, which corresponds to approximately one 1.5-oz shot of 80-proof alcohol (about 14 g alcohol), one 5-oz glass of table wine (11 g), or one 12-oz beer or wine cooler (12.8 g).

(2) FAS is identified by its characteristic facial dysmorphology, growth restriction, and central nervous system/neurodevelopmental abnormalities (Sokol et al. 2003).

(3) Proteomics is the study of how populations of proteins are coded and synthesized in the body, their interactions, their quantification, and their alteration in the presence of illness or other pathology.

References

ANNI, H., AND ISRAEI, Y. Proteomics in alcohol research. Alcohol Research & Health 26(3):219-232, 2002.

BEARER, C.F. Markers to detect drinking during pregnancy. Alcohol Research & Health 25(3):210-218, 2001.

BEARER, C.F.; LEE, S.; SALVATOR, A.E.; ET AL. Ethyl linoleate in meconium: A biomarker for prenatal ethanol exposure. Alcoholism: Clinical and Experimental Research 23:487-493, 1999.

BEARER, C.F.; SALVATOR, A.E.; LEE, S.; ET AL. FAEE in meconium predicts outcome. Alcoholism: Clinical and Experimental Research 24:102A, 2000.

BEARER, C.F.; SALVATOR, A.E.; LEE, S.; ET AL. A comparison of FAEE in meconium to maternal self-reported drinking in predicting neurodevelopment. Pediatric Research 49:183A, 2001.

BEARER, C.F.; JACOBSON, J.L.; JACOBSON, S.W.; ET AL. Validation of a new biomarker of fetal exposure to alcohol. Journal of Pediatrics 143:463-469, 2003.

Centers for Disease Control and Prevention (CDC). Alcohol use among women of childbearing age--United States, 1991-1999. Morbidity and Mortality Weekly Report 51:433-435, 2002.

CHAN, D.; BAR-OZ, B.; PELLERIN, B.; ET AL. Population baseline of meconium fatty acid ethyl esters among infants of nondrinking women in Jerusalem and Toronto. Therapeutic Drug Monitoring 25:271-278, 2003.

CHANG, G. Alcohol-screening instruments for pregnant women. Alcohol Research & Health 25(3):204-209, 2001.

HALMESMAKI, E.; ROINE, R.; AND SALASPURO, M. Gamma-glutamyl transferase, aspartate and alanine aminotransferases and their ratio, mean cell volume and urinary dolichol in pregnant alcohol abusers. British Journal of Obstetrics and Gynaecology 99:278-291, 1992.

KLEIN, J.; KARASKOV, T.; AND KOREN, G. Fatty acid ethyl esters: A novel biologic marker for heavy in utero ethanol exposure: A case report. Therapeutic Drug Monitoring 21:644-646, 1999.

KLEIN, J.; CHAN, D.; AND KOREN, G. Neonatal hair analysis as a biomarker for in utero alcohol exposure. New England Journal of Medicine 347:2086, 2002.

MAY, P.A., AND GOSSAGE, J.P. Estimating the prevalence of fetal alcohol syndrome: A summary. Alcohol Research & Health 25(3):159-167, 2001.

National Institute on Alcohol Abuse and Alcoholism (NIAAA). The Physicians' Guide to Helping Patients With Alcohol Problems. NIH Pub. No. 95-3796. Bethesda, MD: NIAAA, 1995.

NEUHOLD, L.A.; GUO, Q.M.; ALPER, J.; ET AL. High-throughput proteomics for alcohol research. Alcoholism: Clinical and Experimental Research 28:203-210, 2004.

NIEMELA, O.; HALMESMAKI, E.; AND YLIKORKALA, O. Hemoglobin-acetaldehyde adducts are elevated in women carrying alcohol-damaged fetuses. Alcoholism: Clinical and Experimental Research 15:1007-1010, 1991.

ROBINSON, M.K.; MYRICK, J.E.; HENDERSON, L.O.; ET AL. Two-dimensional protein electrophoresis and multiple hypothesis testing to detect potential serum protein biomarkers in children with fetal alcohol syndrome. Electrophoresis 16:1176-1183, 1995.

RUSSELL, M.; MARTIER, S.S.; SOKOL, R.J.; ET AL. Screening for pregnancy risk-drinking. Alcoholism: Clinical and Experimental Research 18:1156-1161, 1994.

RUSSELL, M.; MARTIER, S.S.; SOKOL, R.J.; ET AL. Detecting risk drinking during pregnancy: A comparison of four screening questionnaires. American Journal of Public Health 86:1435-1439, 1996.

SARKOLA, T.; ERKISSON, C.J.; NIEMELA, O.; ET AL. Mean cell volume and gamma-glutamyl transferase are superior to carbohydrate-deficient transferrin and hemoglobin-acetaldehyde adducts in the follow-up of pregnant women with alcohol abuse. Acta Obstetricia et Gynecologica Scandinavica 79:359-366, 2000.

SAVAGE, C.L.; WRAY, J.N.; RITCHEY, P.N.; ET AL. Measurement of maternal alcohol consumption in a pregnant population. Substance Abuse 23:211-214, 2002.

SOKOL, R.J.; DELANEY-BLACK, V.; AND NORDSTROM, B. Fetal alcohol spectrum disorder. JAMA: Journal of the American Medical Association 290:2996-2999, 2003.

STOLER, J.M., AND HOLMES, L.B. Under-recognition of prenatal alcohol effects in infants of known alcohol abusing women. Journal of Pediatrics 135:430-436, 1999.

STOLER, J.M., AND HOLMES, L.B. Recognition of facial features of fetal alcohol syndrome in the newborn. American Journal of Medical Genetics 127C:21-27, 2004.

STOLER, J.M.; HUNTINGTON, K.S.; PETERSON, C.M.; ET AL. The prenatal detection of significant alcohol exposure with maternal blood markers. Journal of Pediatrics 133:346-352, 1998.

STREISSGUTH, A.P.; BARR, H.M.; KOGAN, J.; ET AL. Understanding the Occurrence of Secondary Disabilities in Clients with Fetal Alcohol Syndrome (FAS) and Fetal Alcohol Effects (FAE): Final Report to the Centers for Disease Control and Prevention (CDC). Technical Report No. 96-06. Seattle, WA: University of Washington Publication Services, 1996.

WISNIEWSKI, K.E.; PULLARKAT, R.K.; HARIN, A.; ET AL. Increased urinary dolichol in newborns whose mothers were heavy alcohol users. Annals of Neurology 14:382, 1983.

CYNTHIA F. BEARER, M.D., PH.D., JOAN M. STOLER, M.D., JANINE D. COOK, PH.D., WITH SIRI J. CARPENTER, PH.D.

CYNTHIA F. BEARER, M.D., PH.D., is an associate professor in the Departments of Pediatrics and Neurosciences at Case Western Reserve University, Rainbow Babies and Children's Hospital, University Hospitals of Cleveland, Cleveland, Ohio.

JOAN M. STOLER, M.D., is an assistant professor of pediatrics, Harvard Medical School and Massachusetts General Hospital, Boston, Massachusetts.

JANINE D. COOK, PH.D., is an assistant professor in the Department of Medical and Research Technology, School of Medicine, University of Maryland, Baltimore, Maryland.

SIRI J. CARPENTER, PH.D., is a science editor for Alcohol Research & Health.

This work was supported by grants to Cynthia F. Bearer from the National Institute on Alcohol Abuse and Alcoholism (R01-AA-11839), the National Institute on Environmental Health Science/NIH/EPA (P01-ES-11261), AAMC/CDC/ATSDR Cooperative Agreement (MM-0122-02/02), the Arc of the United States, and the Mary Ann Swetland Center for Environmental Health.

COPYRIGHT 2004 U.S. Government Printing Office
COPYRIGHT 2005 Gale Group