Alcohol and Alcoholism Advance Access originally published online on March 6, 2007
Alcohol and Alcoholism 2007 42(5):407-412; doi:10.1093/alcalc/agm005
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Clinical approach to intestinal maturation in neonates prenatally exposed to alcohol
1 Departament de Bioquímica i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Av/ Diagonal 645, E-08028 Barcelona
2 Unitat d'Alcohologia, Institut Clínic de Psicologia i Psiquiatria, Hospital Clínic, Villarroel 170, E-08036 Barcelona
3 Department of Maternal-Fetal Medicine, Hospital Clínic, Universitat de Barcelona, IDIBAPS, Sabino de Arana 1, E-08028 Barcelona, Spain
* Author to whom correspondence should be addressed at: Departament de Bioquiacutemca i Biologia Molecular, Facultat de Biologia, Universitat de Barcelona, Av/ Diagonal 645, E-08028 Barcelona, Spain. Fax: 3-4021559; E-mail: dolopez{at}ub.edu
Received 21 November 2006; first review notified 15 December 2006; in revised form 24 January 2007; accepted 25 January 2007
 |
ABSTRACT |
|---|
 TOP ABSTRACT IntroductionMethodsResultsDiscussionReferences |
|---|
Aim: The need for a non-invasive diagnosis of the effects of ethanol in utero on the development of the intestine in humans led us to look for a serum marker of the structural integrity of the intestine. We propose apolipoprotein A-IV (apoA-IV) as a possible candidate. In humans this protein is synthesized only by intestinal mucosa, it is expressed in the enterocyte of the foetus from 20 weeks of gestation, and it is released to the blood stream after synthesis. Methods: We measured the levels of apoA-IV in the umbilical cord serum of neonates whose mothers had consumed alcohol during pregnancy and neonates born to women who had not (controls).The gestational age at delivery of the cases studied ranged from 36 to 42 weeks. ELISA and Western blot analysis were used. Results: There was no difference in the mean body weight of neonates from either group. Nevertheless, exposure to ethanol in utero significantly reduced (by about 30%) the apoA-IV levels in serum at birth, regardless of body weight. Conclusion: Our findings suggest that circulating apoA-IV levels could be used as a clinical marker of the prenatal effects of ethanol on the structural integrity of the intestine. Neonatal diagnosis of these intestinal effects could improve post-natal outcome.
|
Introduction |
|---|
|
TOPABSTRACTIntroductionMethodsResultsDiscussionReferences |
|---|
Chronic alcoholism in gestation has a negative impact on the pre- and post-natal development of offspring and causes a series of disorders known as foetal alcohol syndrome (FAS) (Jones et al., 1973
). However, cases with less evident symptoms, referred to as ‘Foetus Alcohol Effects’ (FAE), are included in the proportion of neonates affected by alcohol in utero (Larroque, 1992). Many questions regarding the course of the affected individuals during childhood remained unanswered.
A large number of studies describe the effects of ethanol exposure in utero on the organs of the embryo and foetus (Chaudhuri, 2000). Few experimental studies describe the effects of maternal ethanol consumption on the intestinal development of offspring. Previous studies of rat foetus from our laboratory describe functional, morphological, cellular and molecular disorders of the intestinal mucosa caused by ethanol in utero (López et al., 1989; Estrada et al., 1996a,b, 1998; Montes et al., 1996; Camps et al., 1997). High mortality was observed after the first suckling, which may be attributable to general immaturity of the small intestine at birth López et al., 1989). We demonstrated (Estrada et al., 1996a) that maternal ethanol intake during gestation reduces the number of enterocytes, with compensatory cellular proliferation rates in the intestinal mucosa of offspring at the moment of birth. The effects of ethanol observed in the pre-natal intestine persist after birth and affect growth rate and the capacity of the offspring to absorb nutrients during post-natal development in rats (Buts et al., 1992; Camps et al., 1993; Tavares et al., 1998, 1999; Murillo-Fuentes et al., 2001, 2003; Bhalla et al., 2004).
Ethanol crosses the placenta directly (Abel, 1982) but it is also absorbed rapidly by simple diffusion and passes into the foetal blood. Human foetuses swallow amniotic fluid from mid pregnancy (18–20 weeks gestation)–a physiological phenomenon that gradually increases during pregnancy and stimulates the peristaltic movements of the intestine and bathes the gastric-intestinal system of the foetus until birth.
We previously demonstrated that the ethanol concentration in the amniotic fluid from the stomach of rat foetuses subjected to chronic ethanol exposure is even higher than in the dam's blood (Través et al., 1995). During gestation the foetus does not metabolize ethanol and pregnancy reduces the dam's tolerance (Través and López-Tejero, 1994); consequently, it remains in circulating blood for longer. Therefore, in humans (as in rats) the intestine of neonates whose mothers consume alcohol during gestation may have been exposed directly to ethanol. Studies in human neonates exposed to ethanol during the second half of gestation (Moore et al., 2003) show that meconium (the first fecal matter in neonates) specimens contain fatty acyl ethyl esters (FAEEs), formed by trans-esterification of glycerides with the ethanol. The detection of these ethanol derivatives in meconium indicates the presence of ethanol in the foetal intestine. Moreover, some authors (Brien et al., 2006) have also found increased concentration of FAEEs in the meconium collected from the large intestine of the foetal term guinea pig exposed to ethanol in utero.
There is little information on the effects of ethanol in utero on the human neonate intestine because they are difficult to study prenatally. Only a few clinical cases of evident intestinal disorder in children with Foetal Alcohol Syndrome have been reported: one case was caused by coeliac disease (Hogh and Stenhammar, 1984), another by small-intestine atresia (Tourtet et al., 1997; Vasiliauskas et al., 1997) and in another by abnormal insertion of the umbilical cord and hypertrophic pyloric stenosis (Mangyanda et al., 1998).
However, these effects in rats are well documented. Similar effects to those observed in experimental animals can be expected in human neonates, such as morphological and functional disorders of the intestinal mucosa, which can compromise future enteral feeding, growth and post-natal development. Here we sought a more indirect way to evaluate the cellular damage to the intestinal mucosa of human neonates whose mothers consumed alcohol during gestation. We examined plasma and/or serum levels of apolipoprotein A-IV (apoA-IV), which originates specifically in the intestinal mucosa (Green et al., 1980; Karathanasis and Yunis, 1986), and which could be an indicator of the state of the intestinal mucosa during severe malnutrition in children (El Harim et al., 1993). A clinical condition well characterized by paediatricians is intestinal atrophy in coeliac disease (intolerance to gluten) in children between 2 and 6 years of age (Trier, 1991). We have also found an association between low plasma levels of apoA-IV and this intestinal atrophy (by Western blot in serum samples and immunolocalization in intestinal biopsies, unpublished data).
ApoA-IV is synthesized by enterocytes and is then bound to chylomicra after enteral nutrition and released into the blood stream. However, it is rapidly separated from these lipoproteins and appears in a free form or bound to HDL (Green et al., 1980). ApoA-IV contributes to lipid metabolism (Bisgaier et al., 1985; Stein et al., 1986; Goldberg et al., 1990; Sindelar et al., 1997) and its expression is associated with nutritional changes (Lefevre and Roheim,1984; Sherman and Weinberg, 1988) and lipid intake (Apfelbaum et al., 1987; Kalogeris et al., 1994; Black et al., 1996). In humans this protein is expressed only in the enterocyte of the foetus from week 20 of gestation (Karathanasis and Yunis, 1986), and can therefore be studied in neonates and quantified at birth (Steinmetz et al., 1988; Sánchez-Pozo et al., 1995).
The possible clinical application of Apo A-IV as a marker of the structural integrity of the intestine in the foetus opens up an interesting field in non-invasive diagnosis. Here we measured the serum levels of apoA-IV in arterial cord blood from human neonates whose mothers consumed alcohol during pregnancy (gestational age at delivery ranged from 36 to 42 weeks). To confirm that the neonates were not breastfed immediately after birth, we measured circulating triacylglycerides and total cholesterol. By comparison with neonates born from control mothers, our results indicate that, as in experimental animals, exposure to ethanol in utero also compromises the development of the intestinal mucosa in humans.
|
Methods |
|---|
|
TOPABSTRACTIntroductionMethodsResultsDiscussionReferences |
|---|
Subjects and clinical samples
In 1991, a prospective study was done in the Department of Obstetrics and Gynaecology of the Hospital Clínic to establish the incidence of alcohol consumption and the abuse of illicit drugs during pregnancy (Martínez Crespo et al., 1994
). Mothers who gave birth in the Hospital Clínic in 1991 were included in the study. A standardized questionnaire was administered to all the participants in the first 48 hours after delivery to assess alcohol consumption during pregnancy. Interviews were conducted by two trained research assistants from the Alcohol Unit of our Hospital. The questionnaire was exhaustive and also specifically designed to identify alcohol intake during pregnancy: 1) non consumption, 2) consumption (occasional or habitual, regardless of the amount). Urine samples were routinely collected at some time before delivery. All the urine specimens were screened anonymously for the measurement of drugs and metabolites. Ethanol in urine was screened by alcohol dehydrogenase with a cutoff of 15 mg/L; this method allows positive detection of ethanol only 8–12 h after alcohol consumption. All mothers with positive presence of ethanol in urine were included in the Alcohol group. Some of those mothers declared in the questionnaire that they had consumed alcohol during pregnancy, regardless of the amount (occasional or habitual). Control group was defined as mothers who stated they had consumed no alcohol during pregnancy and who had negative presence of ethanol in urine. In both groups all premature neonates (less than 36 weeks), stillbirths, maternal illicit drug use or HIV-1 or HCV positive were excluded. Serum samples from cord blood were collected and stored at –30°C within 24 h of extraction. Questionnaires, serum and urine samples were coded, thereby ensuring anonymity. After mothers’ selection by questionnaire and urine results, we included 78 women who reported alcohol consumption (alcohol group). The number of participants with no reported alcohol intake during pregnancy and negative presence of ethanol in urine was 139 (control group). Although all patients with delivery at less than 36 weeks were excluded, we also selected two premature cases (35 weeks) from the two groups for Western blot analysis. The study was approved by the Hospital Research Committee and the mothers gave informed consent to their inclusion in the study.
Quantification of apoA-IV in serum by ELISA sandwich
Plasma concentrations of apoA-IV were measured by an ELISA sandwich developed in our laboratory. The diluted samples were pre-treated with a clearing buffer containing cholesterol esterase to eliminate interference from serum lipids. The serum samples and the internal control (for inter- and intra-assays) were diluted 1/10 in phosphate buffer saline PBS and then incubated at 37°C with clearing buffer used for nephelometry which contained: 87.2 g/L aspartic acid, 17.5 g/L sodium cholate, 1.2% Tween 20, 0.4 g/L cholesterol esterase, and 0.1 M Tris, pH 7.7 in (1:1) (v:v). The reaction was halted with ice after 10 min and then PBS-T-BSA (Phosphate Buffer Saline with 0.05% Tween-20, v/v, and of 0.5% Bovine Serum Albumine, w/v) was added, until the final dilution of the sample (1/150 or 1/300) was attained. Clearing buffer was processed without serum as background.
Plates were immunoabsorbed with antibody captor IgG anti-human apoA-IV (500 ng/well), which we obtained in rabbit. Non-specific binding was performed with a blocking solution (PBS with BSA at 0.5% w/v). The standard of apoA-IV (which we purified from human plasma) was diluted in PBS-T-BSA buffer, using a concentration range of 1 to 2000 ng per well. The clearing buffer was not used in standard solutions.
100 µL of the pre-treated samples, the standard, and the pre-treated internal control were incubated for one hour at 37°C. The same antibody bound to biotin (0.5 mg/mL, 12.4 mol biotin/ mol IgG) was used as a detector (100 µL per well, diluted 1/500 in PBS-T-BSA), and incubated during 2 h at 37°C. Afterwards 100 µL of a solution of avidine bound to peroxidase was incubated for 20 min at room temperature. When all the incubation procedures were complete the plate was washed five times with PBS-T (Phosphate Buffer Saline with 0.05% Tween-20, v/v).
Freshly prepared OPD solution (0.4 g/L O-phenyl- endiamine, 7.3 g/L citric acid, 9.6 g/L Na2HSO4 and 0.2 µL H2O2) was used as substrate of the peroxidase. After 20 min at room temperature the reaction was stopped by adding 2.5 M HCl (50 µL/well).
The standard curve was adjusted through polynomic regression analysis. Two dilutions of each sample were assayed (1/150 and 1/300) and the apoA-IV concentration was calculated from the mean of the two dilutions. The dilutions of the sample and the various concentrations of the standard were assayed in duplicate. In each plate we used an internal control in quadruplicate; we obtained from this control the results of intra- and inter-assays (of 4.2% and 9.1% respectively). The plate was read at 492 nm by mean of a ‘Titertek Multiskan’ ELISA reader.
Western blot to detect apoA-IV in serum
The Western blot assays to detect apoA-IV used the proteins separated in 10% Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) and transferred to membranes of ‘Immobilon’ (Millipore®). IgG anti-human apoA-IV obtained in our laboratory from rabbit at 1/500 dilution (with blocking solution) was used as primary antibody; anti-IgG from commercial rabbit obtained in pig (labelled with peroxidase, Horse Radish Peroxidase (HRP)) at 1/15,000 dilution (with blocking solution) was used as secondary antibody (Dako, Glostrup, Denmark). Non-specific binding was performed with a blocking solution (PBS with Bovine Serum Albumin, BSA) at 2% (w/v, BSA/PBS). The Enhanced Cheminal Luminiscence (ECL) commercial system was used (Amersham Pharmacia Biotech, Little Chalfont, Buckinghamshire, England) was used to develop the Western blot.
Triacylglycerides and total cholesterol in serum
The commercial kits ‘TRIG’ and ‘CHOL’ (Medical Analysis Systems, California) were used to measure triacylglycerides and total cholesterol.
Statistical analysis
Results are expressed as mean ± SEM. As the data were not normally distributed, the Mann–Whitney U test was applied. Differences were considered statistically significant at p < 0.05. The statistical package for the Social Sciences (SPSS Inc., Chicago, Illinois), version 10.0 for Windows was used.
|
Results |
|---|
|
TOPABSTRACTIntroductionMethodsResultsDiscussionReferences |
|---|
The age at delivery of the two groups was similar (the medium age was of 28 years; the minimum age was a case of 17 years and the maximum was a case of 43 years). Patients included in the study gave birth between 36 and 42 weeks.
FAS (Jones et al., 1973) defined as the presence of a characteristic pattern of facial anomalies in infants born from mothers with suspected high consumption of alcohol during gestation. In this study, neonates born with FAS were not diagnosed in the alcohol group. There was no difference in the mean body weight of neonates in the two groups (Table 1).
|
ApoA-IV serum levels were significantly lower in neonates from the alcohol group (p < 0.001) than those from the control group (around 30%). No differences were observed in levels of triacylglycerides or total cholesterol (Table 1).
Table 2 shows different parameters in the alcohol group (mothers and neonates) in accordance with the questionnaire and urine results. The percentage of mothers in each of the three groups is similar, and corresponds to approximately one third of the whole alcohol group. There was no difference in the mean body weight or apo A-IV serum levels of neonates born from the three different groups of mothers.
|
ApoA-IV serum levels stratified by neonatal weight are shown in Fig. 1. ApoA-IV serum levels were significantly lower in infants exposed in utero to ethanol when birth weight was less than 3.5 kg. Differences were still present among babies weighing less than 2.5 kg, but they were not statistically significant, because the number of individuals was too small (N = 4).
|
Western blots of apoA-IV, from the serum of neonates from the two groups at different times of gestation (35 and 40 weeks), are shown in Fig. 2. The intensity of the label in the serum of neonates whose mothers consumed alcohol was lower than that of the control group at the same gestational age. In one case (at 40 weeks of gestation) apoA-IV was not detected. These results are in accordance with those obtained by the ELISA analysis (Table 1 and Fig. 1).
|
|
Discussion |
|---|
|
TOPABSTRACTIntroductionMethodsResultsDiscussionReferences |
|---|
The results indicate that apoA-IV levels in neonates whose mothers consumed alcohol during pregnancy are lower than in neonates from control mothers. In general, the mothers in the alcohol group were exposed to moderate amounts of alcohol during pregnancy, as reflected in the weight of the neonates, which was similar in the three groups of mothers (classified by questionnaire responses on ethanol consumption) and also in the fact that this parameter is no different to neonates of control group. Nevertheless, differences in apoA-IV serum levels were significant vs control group in babies weighing less than 3.5 kg at birth.
In humans, the expression of apoA-IV is associated with nutritional changes and lipid intake. In this case, the effects cannot be attributed to differences in lipid intake since the neonates had not suckled and it is supported by the observation that levels of serum triacylglycerides were low in both groups. The serum concentration of apoA-IV from control neonates was similar to the concentrations found in our previous studies (international patent application has been filed, application number EP03738136.5 and publication number EP1560024A1, inventors M.Dolores López Tejero and Carmen Través Polo, title ‘Method of diagnosing changes in the intestinal absorptive surface in an individual’), but below those reported by other authors: 5.7 ± 1.9 mg/100 mL, N = 127 (Steinmetz et al., 1988) and 5.0 ± 2.0 mg/100 mL, N = 5 (Sánchez-Pozo et al., 1995). These differences may be attributed to the methodology used to measure this protein and to the number of individuals tested. The findings reported by Steinmetz included many individuals who were evaluated by immunoelectrophoresis, whilst Sánchez-Pozo included fewer patients, but also used an ELISA sandwich test.
In adults, the broad intestinal effects of chronic and acute ethanol intake are well documented (Baraona et al., 1972; Hoyumpa, 1980; Persson, 1991; Preedy et al., 1993, 1999, Rajendram and Preedy, 2005). After chronic alcohol intoxication, the large intestine of adult rats shows a greater imbalance between enzymes that produce or oxide acetaldehyde, as a result of which acetaldehyde accumulates (Pronko et al., 2002). This mechanism may be produced by local toxicity of ethanol on the intestine and the mucosal damage may activate hyper-regeneration and possibly cause carcinogenesis. The same compensatory cellular proliferation rates were found in intestinal mucosa of rat neonates (Estrada et al., 1996a).
Experimental studies on intestinal exposure to ethanol in utero are more limited. In rat during prenatal development a particular phenotype of the foetal intestine affected by ethanol in utero we found (López et al., 1989; Estrada et al, 1996a,b, 1998; Montes et al., 1996; Camps et al., 1997). This phenotype is characterized by functional and structural disorders of the enterocytes, which affect the organization of the intestinal epithelium and the biosynthetic capacity of digestive enzymes. This anomaly is associated with impaired post-natal development and reduced capacity of offspring to absorb nutrients (Buts et al., 1992; Camps et al., 1993; Tavares et al., 1998, 1999; Murillo-Fuentes et al., 2001, 2003). The decrease in nutrient intake observed during post-natal development could be a consequence of the loss of epithelium resulting in a decline of the number of transport molecules (Bhalla et al., 2004).
However, we found in rat that plasma levels of apoA-IV were unaltered in term foetuses from chronic alcohol mothers (unpublished data), which may be because this protein is also synthesized by the liver in rat; in humans it is only synthesised in the intestine. This difference could mask alterations of blood levels of apo A-IV in the rat.
In humans few studies have addressed foetal intestinal disorders caused by ethanol in utero. This study examined a group of mothers who consumed alcohol during pregnancy, but the goal was not to assess the exact pattern of ethanol consumption in mothers. It was difficult to obtain a reliable answer, as shown by, the results of percentage of individuals in Table 2. However, the intake of ethanol did not have severe effects on the body development of the foetuses, as neonate weight did not differ between the two clinical groups. This observation could be explained because little ethanol was consumed and because the mothers did not suffer from severe malnutrition. Moreover, other factors which may contribute to the birth- weight effects, as the smoking and the use of other drugs during pregnancy, whose we have been conveniently checked in this study (see exclusion criteria in methods).
Even at a low intake, alcohol can have adverse effects on neonates and lead to problems in childhood (Streissguth et al., 1990; Sood et al., 2001). Many studies have addressed the relationship between ethanol consumption during pregnancy and neonate weight. About half of these report significant correlation between the two, whereas the other half find no relationship (Larroque, 1992). Given that few studies have examined growth related to moderate alcohol consumption during pregnancy, the evidence so far is inconclusive (Forrest et al., 1992).
This study is the first in humans to describe an alteration in the serum levels of a protein that originates in the intestine (apoA-IV), and which may be linked to a disorder or underdevelopment in the intestinal epithelium, resulting in the loss and/or reduction of number of enterocytes. This effects observed in the intestine of foetus exposed to etanol in utero, may be caused by the interaction of this toxic with enterocytes (by a direct and/or indirect way). In vitro studies show apoptosis in human intestinal cells induced by acute and low concentration of ethanol (Asai et al., 2003). The cellular mechanisms of pathogenesis could be related to oxidative stress and reactive aldehyde production (Henderson et al., 1999), with the reduction of free retinoic acid levels in foetus (Zachman and Grummer, 1998) (a potent mediator in embryogenesis and differentiation), or with alteration of growth regulatory factors (Chaudhuri, 2000). Some of the latter, modulators of cell growth and function, may be related with the decreased intestinal motility and pseudo-obstruction observed in some children with FAS, which may be caused by changes in the musculature and neurons in the gastrointestinal tract.
We conclude that exposure to ethanol in utero causes alterations of the intestinal mucosa in humans, as in rats. This is reflected by the decrease in serum apoA-IV concentration, a protein synthesized solely by enterocytes. Further studies are needed. If this hypothesis is to be confirmed, these studies should correlate morphological, biochemical and cytological parameters of the foetal intestine with apoA-IV serum or plasma levels. During late pregnancy to perform experiments and to sample foetal or neonate intestines, it is clearly difficult in humans. Here we present a simple, non-invasive method to diagnose intestinal atrophy or disorder at birth. This serum marker is an essential tool, especially in neonatal and paediatric clinics, as intestinal anomalies in the first months of life condition the future development of the baby.
|
ACKNOWLEDGEMENTS |
|---|
This work was partially supported by a grant from the Fondo de Investigaciones Sanitarias from the Spanish Heath Ministery (93/0551). Carmen Través Polo was the recipient of a fellowship from the Spanish Heath Ministery. The authors wish to thank the medical staff (Dr. Aureli Torné and Dr. Josep M. Martinez) and the nursing staff (Esther Rebull and Ana Carrión) of the Institut Clínic de Ginecologia, Obstetrícia i Neonatologia for their very valuable assistance. We also thank the Language Advisory Service at the University of Barcelona for revising the English manuscript.
|
References |
|---|
|
TOPABSTRACTIntroductionMethodsResultsDiscussionReferences |
|---|
Abel E. L. Consumption of alcohol during pregnancy: a review of effects of growth and development of offspring. Human Biology (1982) 54:421–453.[Web of Science][Medline]
Apfelbaum T. F., Davidson N. O., Glickman R. M. Apolipoprotein synthesis in rat intestine: regulation by dietary triglyceride. American Journal of Physiology (1987) 252:G662–G666.[Web of Science][Medline]
Asai K., Buurman W. A., Reutelingsperger C. P. M., et al. Low concentrations of ethanol induce apoptosis in human intestinal cells. Scandinavian Journal of Gastroenterology (2003) 38:1154–1161.[CrossRef][Web of Science][Medline]
Baraona E., Pirola R. C., Lieber C. S. Small intestinal damage and changes in cell population produced by ethanol ingestion in the rat. Gastroenterology (1972) 66:226–234.
Bhalla S., Mahmood S., Mahmood A. Effect of prenatal exposure to ethanol on postnatal development of intestinal transport functions in rats. European Journal of Nutrition (2004) 43:109–115.[CrossRef][Web of Science][Medline]
Bisgaier C. L., Sachdev O. P., Megna L., et al. Distribution of apolipoprotein A-IV in human plasma. Journal of Lipid Research (1985) 26:11–25.[Abstract]
Black D. D., Wang H., Hunter F., et al. Intestinal expression of apolipoprotein A-IV and C-II is coordinately regulated by dietary lipid in newborn swine. Biochemical and Biophysical Research Communications (1996) 221:619–624.[CrossRef][Web of Science][Medline]
Brien J. F., Chan D., Green C. R., et al. Chronic prenatal ethanol exposure and increased concentration of fatty acid ethyl esters in meconium of term fetal guinea pig. Therapeutic Drug Monitoring (2006) 28:345–350.[CrossRef][Web of Science][Medline]
Buts J. P., Sokal E. M., Van Hoof F. Prenatal alcohol exposure to ethanol in rats, effects on postnatal maturation of the small intestine and liver. Pediatric Research (1992) 32:574–579.[Web of Science][Medline]
Camps L., Estrada G., López-Tejero M. D. Desarrollo intestinal en la descendencia de madres alcohólicas. Actualidad Nutricional (Milupa, Madrid) (1993) 16:26–34.
Camps L., Kédinger M., Simon-Assmann P., et al. Effect of prenatal exposure to ethanol on intestinal development of rat fetuses. Journal of Pediatric Gastroenterology and Nutrition (1997) 24:302–311.[CrossRef][Web of Science][Medline]
Chaudhuri J. D. Alcohol and the developing fetus, a review. Medical Science Monitor (2000) 6:1031–1041.[Medline]
El Harim I., Befort J. J., Balafrej A., et al. Lipids and lipoproteins of malnourished children during early renutrition: apolipoprotein A-IV as a potential index of recovery. American Journal of Clinical Nutrition (1993) 58:407–411.
Estrada G., Del Río J. A., García-Valero J., et al. Ethanol in utero induces epithelial cell damage and altered kinetics in the developing rat intestine. Teratology (1996a) 54:245–254.[CrossRef][Web of Science][Medline]
Estrada G., Krasinski S. D., Rings E. H. H. M., et al. Prenatal ethanol exposure alters the expression of intestinal hydrolases mRNAs in newborn rats. Alcoholism-Clinical and Experimental Research (1996b) 20:1662–1668.[CrossRef]
Estrada G., Krasinski S. D., Grand R. J., et al. Defective intracellular processing of lactase-phorizin hydrolase protein in rats prenatally exposed to ethanol. Alcoholism-Clinical and Experimental Research (1998) 22:1177–1183.
Forrest F., DuFlorey C. V., Taylor D. Maternal Alcohol Consumption and child development. International Journal of Epidemiology (1992) 21:S17–S23.[Abstract]
Goldberg I. J., Scheraldi C. A., Yacoub L. K., et al. Lipoprotein Apo C-II activation of lipoprotein lipase. Journal of Biological Chemistry (1990) 265:4266–2477.
Green P. H. R., Glickman R. M., Riley J. W. Human apolipoprotein A-IV. Intestinal origin and distribution in plasma. Journal of Clinical Investigation (1980) 65:911–919.[Web of Science][Medline]
Henderson G. I., Chen J. J., Schenker S. Ethanol, oxidative stress, reactive aldehydes, and the fetus. Frontiers in Bioscience (1999) 4:541–550.[CrossRef]
Hogh B., Stenhammar L. Coeliac disease coexistent with fetal alcohol syndrome. European Journal of Pediatrics (1984) 143:74–75.[Web of Science][Medline]
Hoyumpa A. M. Mechanisms of thiamine deficiency in chronic alcoholism. American Journal of Clinical Nutrition (1980) 33:2750–2761.
Jones K. L., Smith D. W., Ulleland C. N., et al. Pattern of malformation in offspring of chronic alcoholic mothers. Lancet (1973) 9:1267–1271.
Kalogeris T., Fukagawa K., Tso P. Synthesis and lymphatic transport of intestinal apolipoprotein A-IV in response to graded doses of triglyceride. Journal of Lipid Research (1994) 35:1141–1151.[Abstract]
Karathanasis S. K., Yunis I. Structure, evolution, and tissue-specific synthesis of human apolipoprotein A-IV. Biochemistry (1986) 25:3962–3970.[CrossRef][Medline]
Larroque B. Alcohol and the fetus. International Journal of Epidemiology (1992) 21(Suppl. 1):S8–S16.[Abstract]
Lefevre M., Roheim P. S. Metabolism of apolipoprotein A-IV. Journal of Lipid Research (1984) 25:1603–1610.[Web of Science][Medline]
López D., Arilla E., Colás B., et al. Low intestinal lactase activity in offspring from ethanol-treated mothers. Biology of the Neonate (1989) 55:204–213.[Web of Science][Medline]
Mangyanda M. L., Mbuila C., Geniez L., et al. Fetal alcohol syndrome and hypertrophic pyloric stenosis in tow brothers. Archives de Pediatrie (1998) 5:695–696.[CrossRef][Web of Science][Medline]
Martínez Crespo J. M., Antolín E., Comas C., et al. The prevalence of cocaine abuse during pregnancy in Barcelona. European Journal of Obstetrics Gynecology and Reproductive Biology (1994) 56:165–167.[CrossRef][Web of Science][Medline]
Montes J. F., Estrada G., López-Tejero M. D., et al. Changes in the enterocyte cytoskeleton in newborn rats exposed to ethanol in utero. Gut (1996) 38:846–852.
Moore C., Jones J., Lewis D., et al. Prevalence of fatty acid ethyl esters in meconium specimens. Clinical Chemistry (2003) 49:133–136.
Murillo-Fuentes M. L., Artillo R., Carreras O., et al. Effects of maternal chronic alcohol administration in the rat: lactation performance and pup's grown. European Journal of Nutrition (2001) 4:147–154.
Murillo-Fuentes M. L., Murillo M. L., Carreras O. Effects of maternal etanol consumption during pregnancy or lactation on folic acid intestinal absorption in suckling rats. Life Sciences (2003) l73:219–2209.
Persson J. Alcohol and the small intestine. Scandinavian Journal of Gastroenterology (1991) 26:3–15.[Web of Science][Medline]
Preedy V. R., Marway J. S., Siddiq T., et al. Gastrointestinal protein turnover and alcohol misuse. Drug and Alcohol Dependence (1993) 34:1–10.[Medline]
Preedy V. R., Reilly M. E., Patel V. B., et al. Protein metabolism in alcoholism: effects on specific tissues and whole body. Nutrition (1999) 15:604–608.[CrossRef][Web of Science][Medline]
Pronko P., Bardina L., Satanovskaya V., et al. Effect of chronic alcohol consumption on the ethanol- and acetaldehyde-metabolising systems in the rat gastrointestinal tract. Alcohol and Alcoholism (2002) 37:229–235.
Rajendram R., Preedy V. R. Effect of alcohol consumption in the gut. Digestive Disease (2005) 23:214–221.[CrossRef]
Sánchez-Pozo A., Ramírez M., Gil A., et al. Dietary nucleotides enhance plasma lecithin cholesterolacyl transferase activity and apolipoprotein A-IV concentrations in pre-term newborn infants. Pediatric Research (1995) 37:328–333.[Web of Science][Medline]
Sherman J. R., Weinberg R. B. Serum apolipoprotein A-IV and lipoprotein cholesterol in patients undergoing total parenteral nutrition. Gastroenterology (1988) 95:394–401.[Web of Science][Medline]
Sindelar P. J., Chonjnacki T., Valtersson C. Role of apolipoprotein A-IV in Hepatic Lipase-catalized dolichol acylation and phospolypid hydrolysis. Biochemistry (1997) 36:1807–1813.[CrossRef][Medline]
Sood B., Delaney-Black V., Covington C., et al. Prenatal alcohol exposure and childhood behavior at age 6 to 7 years: I. Dose-response effect. Pediatrics (2001) 108:1–9.
Stein O., Stein Y., Lefevre M., et al. The role of apolipoprotein A-IV in reverse cholesterol transport studied with cultured cells and liposomes derived from an esther along of phosphatidylcholine. Biochimica et Biophysica Acta (1986) 878:7–13.[Medline]
Steinmetz A., Czekelius P., Thiemann E., et al. Changes of apolipoprotein A-IV in the human neonate: evidence for different inductions of apolipoproteins A-IV and A-I in the postpartum period. Atherosclerosis (1988) 69:21–27.[CrossRef][Web of Science][Medline]
Streissguth A. P., Barr H. M., Sampson P. D. Moderate prenatal alcohol exposure: on child IQ and learning problems at age 7 1/2 years. Alcoholism-Clinical and Experimental Research (1990) 14:662–669.[CrossRef]
Tavares E., Carreras O., Gomez-Tubio A., et al. Zinc intestinal absorption in newborn rats at 21 day postpartum: effects of maternal ethanol consumption. Life Sciences (1998) 62:787–797.[CrossRef][Web of Science][Medline]
Tavares E., Gómez-Tubio A., Murillo M. L., et al. Folic acid intestinal absorption in newborn rats at 21 day postpartum: effects of maternal ethanol consumption. Life Sciences (1999) 22:2001–2010.
Tourtet S., Michaud L., Gottrand F., et al. Small intestine atresia and abnormal insertion of the umbilicus in a child with fetal alcohol syndrome. Archives de Pediatrie (1997) 4:650–652.[CrossRef][Web of Science][Medline]
Través C., López-Tejero M. D. Ethanol elimination in alcohol-treated pregnant rats. Alcohol and Alcoholism (1994) 29:385–395.
Través C., Camps L., López-Tejero M. D. Liver alcohol deshydrogenase activity and ethanol levels during chronic ethanol intake in pregnant rats and their offspring. Pharmacology Biochemistry and Behavior (1995) 52:93–99.[CrossRef][Web of Science][Medline]
Trier J. S. Celiac sprue. New England Journal of Medicine (1991) 325:1709–1719.[Web of Science][Medline]
Vasiliauskas E., Piccolo D. A., Flores A. F., et al. Chronic intestinal pseudoobstruction associated with fetal alcohol syndrome. Digestive Diseases and Sciences (1997) 42:1163–1167.[CrossRef][Web of Science][Medline]
Zachman R. D., Grummer M. A. The interaction of ethanol and vitamin A as a potential mechanism for the pathogenesis of Fetal Alcohol Syndrome. Alcoholism-Clinical and Experimental Research (1998) 22:1544–1556.
CiteULike 
Connotea 
Del.icio.us What's this?
| ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||