Serum bile acids and their conjugates in breast-fed infants with prolonged jaundice

Serum bile acids and their conjugates were analysed in 20 breast-fed infants with prolonged jaundice. The mean total bile acid levels in serum were increased in the breast-fed infants with jaundice, as compared with those in either breastor bottle-fed infants without jaundice. However, there were no significant differences between the groups. All the breast-fed infants examined, regardless of association with jaundice, had a bile acid pattern dominated by taurine conjugates (the ratio of glycine- to taurine-conjugated bile acid, G/T ratio, less than 1.00). In contrast, the bottle-fed infants without jaundice had a pattern dominated by glycine conjugates (G/T ratio, more than 1.00). Among the breast-fed infants with jaundice, the mean G/T ratio in those who had serum bilirubin levels over 10 mg/100 ml was significantly lower than that in those who had serum bilirubin levels of less than 10 mg/100 ml. The altered bile acid metabolism might be associated with the pathology of breast milk jaundice.Abbreviation LP-X lipoprotein-X     

Abnormal results of biochemical liver function tests in breast-fed infants with prolonged indirect hyperbilirubinaemia

To clarify the relationship between hyperbilirubinaemia and abnormal results of biochemical liver function tests in infants with breast milk jaundice (BMJ), 58 breast-fed infants with indirect hyperbilirubinaemia were enrolled in this study. Sera obtained from the above infants were subjected to routine liver function tests. Although serum transaminases were within normal limits in all 58 patients, serum alkaline phosphatase levels were abnormally increased in 13, gamma-glutamyltranspeptidase in 8 and total bile acids in 11 out of all patients examined. A total of 18 (31%) patients had abnormal results in at least one item of the liver function tests. The intrinsic bile acid loading test showed postprandial increases in bile acids in 5 of 16 (31%) patients examined at either 60 or 120 min, while all 13 breast-fed, agematched controls had no abnormal results. The decrease in rate of serum bilirubin levels after the 3-day discontinuation of breast-feeding was significantly less in patients with increased fasting bile acids than in patients with normal fasting levels of serum bile acids. These results may suggest that mild hepatic dysfunction or cholestasis is associated with indirect hyperbilirubinaemia in some infants with BMJ.     

Free fatty acids in the development of breast milk jaundice

The incubation of milk, at 4 degrees C, from mothers of infants with breast milk jaundice (BMJ) is reported to result in significantly higher levels of free fatty acids (FFA) compared with milk from controls. Single milk samples collected under standard conditions were obtained from four mothers of infants with BMJ and 14 control donors matched for stage of lactation. Milk samples were analyzed for the concentrations of FFA, using thin-layer gas chromatographic techniques. In addition, serum total fatty acids were measured in mothers and infants. The concentrations of FFA increased after storage of the milk from both the jaundiced and control groups. No differences were observed in the composition of milk FFA before or after incubation, when respective values were compared between these two groups. Similarly, no differences were detected in serum total fatty acids in either infants or mothers. The observation that increased levels of FFA in milk are associated with BMJ was not confirmed.     

A study of the relationship between bile salts, bile salt-stimulated lipase, and free fatty acids in breast milk: normal infants and those with breast milk jaundice

Breast milk jaundice has been reported to be associated with increased lipase activity and elevated free fatty acid (FFA) concentrations within breast milk. We have previously shown that bile salts are present in small concentrations in breast milk and the aim of this study was to examine the relationship of bile salt-stimulated lipase (BSSL) activity, FFA concentration, and bile salt concentration in milks of normal infants and the milk of infants with breast milk jaundice. Mothers of healthy newborn infants were recruited in the early newborn period and 42 provided breast milk samples at 2 weeks, 30 at 6 weeks, 16 at 10 weeks, and 13 at 14 weeks postnatally. We initially studied the effect of lactation on bile salts and found there was a significant decline in both cholate and chenodeoxycholate levels with duration of lactation (p less than 0.05). There was also a significant fall in BSSL activity with duration of lactation (p less than 0.05), but no correlation was found between BSSL activity and bile salt concentration. FFA concentrations were similar throughout lactation and were not related to either BSSL activity or bile salt concentration. There was a significant increase in the concentration of cholate and the cholate-to-chenodeoxycholate ratio in the milks of 12 infants with breast milk jaundice compared with normal milks, the BSSL activity was similar and contrary to previous reports, the FFA concentration was not increased in the milks of infants with breast milk jaundice.     

Urinary phenolic acid and alcohol excretion in the newborn.

Mean urinary excretion values of some phenolic acids and alcohols have been measured by gas chromatography in 44 neonates (36 males, 6 females) during the first 2 days and days 3-7 of life, and the effect of prematurity and jaundice assessed. 4-Hydroxy-3-methoxymandelic acid (VMA) output rises immediately after birth in term but not in preterm infants. A similar increase in homovanillic acid (HVA) output was restricted to nonjaundiced term babies; in nonjaundiced preterm babies there was a steady rise during the first week. The ratio of HVA to VMA output was higher in these infants than in adults, suggesting a more rapid turnover of dopamine than adrenaline and noradrenaline. Unlike adult values, both HVA and VMA excretion values were directly related to urine volume, an observation perhaps related to renal immaturity. An unexplained reduction in HVA output in jaundiced as opposed to nonjaundiced infants was observed in the first 2 days of life. The ratio of 4-hydroxy-3-methoxyphenylglycol to VMA was about the same as in the adult. p-Hydroxyphenyl-lactic acid (p-HPLA), because of its superior stability, was measured in preference to p-hydroxyphenylpyruvic acid as an index of tyrosyluria. An output of 1 mg p-HPLA/24 h is proposed as the upper limit of normal. Prematurity was associated with a significant rise in p-HPLA output. A dramatic increase in excretion of this acid was noted in jaundiced, compared with nonjaundiced infants, presumably a manifestation of general enzyme immaturity.     

Effect of milk feeding on intestinal bilirubin absorption in the rat

The hypothesis that the etiologic mechanism of the late-onset, prolonged, unconjugated hyperbilirubinemia of the breast-fed infant, known as the breast milk jaundice syndrome, results from exaggeration of intestinal bilirubin absorption has been investigated in an adult rat model, which permits quantitative measurement of the enterohepatic circulation of bilirubin. After instillation of unconjugated bilirubin in buffer into the duodenum, 25% of the dose was absorbed and appeared in bile. Administration of bilirubin in human milk or cow milk formula resulted in a marked reduction in absorption to 2%. Administration of bilirubin in milk from mothers of infants with breast milk jaundice syndrome not only failed entirely to prevent the absorption of bilirubin, but enhanced late absorption, to produce a total absorption of 60% of the bilirubin dose. Thus, although normal milk significantly retarded intestinal bilirubin absorption and diminished the bilirubin load to the liver, milk from mothers of infants with breast milk jaundice syndrome appeared to enhance the enterohepatic circulation of bilirubin and to increase the total hepatic bilirubin load. This exaggeration of the enterohepatic circulation of bilirubin may be related to the increased concentrations in these milks of long-chain nonesterified fatty acids.     

ANTIBODY PRODUCTION BY THE MAMMARY GLAND IN MOTHERS AFTER ARTIFICIAL ORAL COLONISATION OF THEIR INFANTS WITH A NON-PATHOGENIC STRAIN E. COLI 083

Abstract. Twenty five breast-fed and 25 formula-fed infants were colonised by oral administration of a living suspension of E. coli 083. Twenty breast-fed and 13 formula-fed infants were followed as controls. Specific antibody titres in serum, stool filtrates and milk, and secretory IgA levels in stool filtrates and milk were determined in samples taken fortnightly from birth until 20 weeks of age. The haemagglutinating antibody in serum and milk increased in the colonised groups, but in stool filtrates an inhibitory effect of breast-milk was demonstrated. Secretory IgA levels in stool filtrates were significantly higher in colonised infants and breast-fed controls than in bottle-fed infants during the period of breast feeding. Then levels in the colonised groups remained high, but in breast-fed controls they decreased to values found in bottle-fed controls. Artificial colonisation evoked local antibody and secretory IgA responses in the intestine, as well as an antibody response in the mother's mammary gland. The possible protective effect of those responses is discussed.     

Antibody production by the mammary gland in mothers after artificial oral colonisation of their infants with a non-pathogenic strain E. coli 083.

Twenty five breast-fed and 25 formula-fed infants were colonised by oral administration of a living suspension of E. coli 083. Twenty breast-fed and 13 formula-fed infants were followed as controls. Specific antibody titres in serum, stool filtrates and milk, and secretory IgA levels in stool filtrates and milk were determined in samples taken fortnightly from birth until 20 weeks of age. The haemagglutinating antibody in serum and milk increased in the colonised groups, but in stool filtrates an inhibitory effect of breast-milk was demonstrated. Secretory IgA levels in stool filtrates were significantly higher in colonised infants and breast-fed controls than in bottle-fed infants during the period of breast feeding. Then levels in the colonised groups remained high, but in breast-fed controls they decreased to values found in bottle-fed controls. Artificial colonisation evoked local antibody and secretory IgA responses in the intestine, as well as an antibody response in the mother's mammary gland. The possible protective effect of those responses is discussed.     

Breast milk jaundice; the role of lipoprotein lipase and the free fatty acids

Lipoprotein lipase activity and free fatty acid concentrations were measured in samples of milk collected from mothers of infants without and with prolonged neonatal jaundice. The lipoprotein lipase and free fatty acid values in the milk from mothers of infants without jaundice were found to increase with the duration of breast-feeding until the 12th postpartum day, and then to fall to the original levels. In the group of mothers with jaundiced infants both lipoprotein lipase and free fatty acid values were found within normal limits when measured between 15th and 37th days post-partum.These findings indicate that increased values of lipoprotein lipase and free fatty acids in the milk are not responsible for the development of breast-milk jaundice.     

How accurate are neonatologists in identifying clinical jaundice in newborns?

This study reevaluates the clinical ability to accurately identifyjaundice in neonates. Three hundred seventy-one term infants were clinically asseseed forjaundice, before discharge home on day 2 to 3 of life. Bilirubin levels obtained at the same time were significantly higher in the newborns clinically diagnosed as beingjaundiced. Our neonatologists were able to diagnose jaundice at clinically low levels, and not to misdiagnose significant hyperbilirubinemia in the majority of the infants. The trained human eye can still discriminate between the jaundiced and nonjaundiced newborn, and clinical impression of jaundice remains a reliable primary screening tool for significant neonatal hyperbilirubinemia.     

A new breast milk supplement for preterm infants

Objective : To assess the effect of a new formula (Prenan), which contains n-3 and n-6 long-chain polyunsaturated fatty acids (LC PUFA) on the fatty acid profile of preterm infants.
Methodology : Plasma fatty acids were measured in 61 preterm infants at term by gas liquid chromatography. In 20 of these infants, paired samples were collected and changes in fatty acids with time analysed.
Results : Plasma docosahexaenoic acid (DHA) levels were higher in those who had been fed expressed breast milk (EBM) ±/or Prenan compared with those fed standard formula ± EBM, P <0.05. The plasma arachidonic acid (AA) levels of infants fed Prenan were not different to those fed EBM, both groups achieving higher levels than infants fed standard formula, P <0.05. Further, paired analysis demonstrated that DHA levels increased in infants changed from standard formula to Prenan to levels equal or higher than those of fully breast-fed infants ( P <0.01), whereas DHA levels remained unchanged with time in all other groups.
Conclusions : The fatty acid composition of Prenan enables preterm infants fed formula to have plasma DHA and AA levels similar to those of infants fed breast milk and consequently different to those of infants fed standard formula. Prenan is an appropriate supplement to breast milk for preterm infants in that it provides LC PUFA as well as additional phosphorus and protein without exposing the infant to intact cows milk protein.     

Unconjugated, glycine-conjugated, taurine-conjugated bile acid nonsulfates and sulfates in urine of young infants with cholestasis

A direct assay system for conjugated bile acids using an enzymatic procedure and high-performance liquid chromatography was used for the analysis of urinary bile acid profiles in young infants with intrahepatic cholestasis (idiopathic neonatal hepatitis syndrome) or extra-hepatic biliary atresia. The major urinary bile acids were cholate and chenodeoxycholate conjugates, but a small amount of deoxycholate and 3 beta-hydroxy-5-cholenate conjugates were detected. Although there was no significant difference in total bile acid excretion between patients with intrahepatic cholestasis and extrahepatic biliary atresia, mean ratios of cholate to chenodeoxycholate and sulfated to total urinary bile acids were different between the two groups examined (5.63 +/- 2.83 vs. 2.50 +/- 1.25, p less than 0.05, 15.8 +/- 9.9 vs. 34.5 +/- 9.9%, p less than 0.005). The proportion of taurine-conjugated chenodeoxycholate in the sulfate fraction to the total bile acid was lower in intrahepatic cholestasis, compared with that in biliary atresia (7.7 +/- 7.5 vs 22.7% +/- 7.8%, p less than 0.005). The greater ratio of cholate to chenodeoxycholate and the reduced excretion of sulfated urinary bile acids in intrahepatic cholestasis was due to decreased taurine-conjugated chenodeoxycholate sulfate excretion.     

EXCRETION OF BILE ACIDS IN ERYTHROBLASTOSIS FOETALIS

The excretion pattern of intramuscularly injected cholic acid-24–¹⁴C was studied for 4 days after the injection in 10 cases of erythro-blastosis (EB). Seven patients with EB and raised serum conjugated bilirubin excreted 3643% of the injected isotope in the urine, whereas the amounts of isotope in the faeces varied greatly. In 3 cases without raised serum conjugated bilirubin less isotope was recovered in the urine and always more than 10% of injected isotope was recovered in the faeces. Cholic acid-24–¹⁴C was excreted essentially unchanged in all cases but in conjugated form. In all cases of EB the urine was found to contain bile acids, chiefly cholic acid. The infants with EB associated with cholestasis excreted 4.8–132.3 μmol of these acids per day; the corresponding values in the absence of cholestasis being 0.4–0.9 μmol per day. In the infants with physiological jaundice the excretion ranged from less than 0.01 to 0.7 μmol per day; the correspondign values in the 2 patients with hyperbilirubinaemia were about 0.2 μmol per day. The infants with EB associataed with cholestasis were found to excrete as large amounts of bile acids in the urine as the infants with intrahepatic cholestasis. These findings strongly suggest that increased serum conjugated bilirubin, irrespective of the patho-genesis of the liver damage, is associated with an impaired bile acid excretion to the intestine. EB without increased serum conjugated bilirubin did not seem to alter the bile acid metabolism, since the urinary excretion of cholic acid and chenodeoxycholic acid in these cases was practically the same as in jaundiced newborn infants.     

Fatty acid composition of the milk lipids of Fulani women and the serum phospholipids of their exclusively breast-fed infants

We previously reported that, relative to milk of women elsewhere in the world, the lipid fraction of milk of Fulani women in northern Nigeria contained relatively low proportions of alpha-linolenic acid and docosahexaenoic acid (DHA). This led us to question the essential fatty acid status of Fulani infants and the relation between the proportion of critical n-3 and n-6 fatty acids in the serum phospholipids of the mothers, their milk, and the serum phospholipids of their exclusively breast-fed infants. We were also interested in the effect de novo intermediate chain length-fatty acids (C10-C14) had on the proportions of critical and non-essential fatty acids in milk. Capillary gas-liquid chromatography was used to analyze the fatty acid content of the total milk lipids of 34 Fulani women, as well as the fatty acid content of serum phospholipids of the women and their breast-fed infants during the first 6 months of life. The proportions of critical n-3 and n-6 fatty acids in the milk of the Fulani women were adequate, but the proportions of these same fatty acids were low in their exclusively breast-fed infants. The serum phospholipids of the infants contained 18.8% linoleic acid, 0.13% alpha-linolenic acid, 12.8% arachidonic acid, and 3.40% DHA, whereas, the mean percentages of linoleic, alpha-linolenic, arachidonic and DHA in the serum phospholipids of the Fulani mothers' were 21.4, 0.20, 9.79, and 1.97, respectively. There was a strong positive correlation between fatty acid content of serum phospholipids of Fulani women and the fatty acid content of their milk lipids. As the proportion of C10-C14 fatty acids in the milk lipids increased, the proportions of critical n-3 and n-6 fatty acids in milk remained relatively constant; however, proportions of three non-essential fatty acids decreased dramatically. C10-C14 fatty acids do not appear to displace critical n-3 and n-6 fatty acids in milk.     

Investigation of serum bile acids; seven patients with Alagille syndrome

To clarify whether an abnormal bile acid pattern has a role in the pathogenesis of Alagille syndrome, we compared serum bile acid patterns in seven with Alagille syndrome with those of patients with congenital biliary atresia (CBA), neonatal hepatitis (NH) and normal infants.Of the seven patients with Alagille syndrome, four patients were younger and three were older than 1 year. The mean total serum bile acid level in the infants was higher than in older subjects. There was a dissociation between the levels of serum total bile acid and bilirubin in three of the seven cases. The mean total bile acid levels in serum were in the following decreasing order: CBA, Alagille syndrome, NH and controls.The ratio of cholate to chenodeoxycholate in the younger patients with Alagille syndrome was significantly higher than CBA (P<0.001). However, no specific bile acid pattern was found in Alagille syndrome by high-performance liquid chromatography (HPLC).Abbreviations TBA total bile acids - FBA free bile acids - conj-BA conjugated bile acids - C/CDC ratio of cholate to chenodeoxycholate - G/T ratio of glycine conjugates to taurine conjugates - GPT glutamic pyruvic transaminase - CBA congenital biliary atresia - NH neonatal hepatitis - HPLC high performance liquid chromatography - GCA glycocholate - TCA taurocholate - GCDCA glycochenodeoxycholate - TCDCA taurochenodeoxycholate     

Unconjugated, Glycine-conjugated, Taurine-conjugated Bile Acid Nonsulfates and Sulfates in Urine of Young Infants with Cholestasis

ABSTRACT. A direct assay system for conjugated bile acids using an enzymatic procedure and high-performance liquid chromatography was used for the analysis of urinary bile acid profiles in young infants with intrahepatic cholestasis (idiopathic neonatal hepatitis syndrome) or extra-hepatic biliary atresia. The major urinary bile acids were cholate and chenodeoxycholate conjugates, but a small amount of deoxycholate and 3β-hydroxy-5-cholenate conjugates were detected. Although there was no significant difference in total bile acid excretion between patients with intrahepatic cholestasis and extrahepatic biliary atresia, mean ratios of cholate to chenodeoxycholate and sulfated to total urinary bile acids were different between the two groups examined (5.63±2.83 vs. 2.50±1.25, p <0.05, 15.8±9.9 vs. 34.5±9.9%, p < 0.005). The proportion of taurine-conjugated chenodeoxycholate in the sulfate fraction to the total bile acid was lower in intrahepatic cholestasis, compared with that in biliary atresia (7.7±7.5 vs. 22.7±7.8 %, p < 0.005). The greater ratio of cholate to chenodeoxycholate and the reduced excretion of sulfated urinary bile acids in intrahepatic cholestasis was due to decreased taurine-conjugated chenodeoxycholate sulfate excretion.     

Age and diet effects on fecal bile acids in infants

Fecal bile acid patterns and concentrations have been determined for 28 infants who were followed from average ages of 3-11 months. Half were solely breast-fed and half were solely formula-fed at the beginning of the study. Breast-fed infants were found to have significantly (p less than 0.05) lower concentrations of cholic acid than the formula-fed group, up to an average age of 5 months. Concentrations of deoxycholic and lithocholic acids were directionally lower in breast-fed infants at all ages. Concentrations of chenodeoxycholic acid were similar for both groups throughout the study. At the end of the study, breast-fed infants were excreting 17% of their total bile acids in the form of secondary acids, compared to 33% for formula-fed infants. This pattern persisted long after the infants began weaning. Formula-fed infants were found to have lithocholic acid in their stools at a significantly (p less than 0.05) earlier age than breast-fed infants. Appearance of deoxycholic acid was at similar ages for both groups. Both of these secondary acids were found to occur at much younger ages (approximately 2 months) than has been previously reported. These observed differences are attributed to the distinct intestinal microbial populations encouraged by the different diets.     

Neonatal Jaundice and Fatty Acid Composition of the Maternal Diet

ABSTRACT. The role of serum fatty acid composition in neonatal jaundice was studied by comparing the incidence of jaundice among 332 newborn infants receiving breast milk from mothers on a diet with either a low (0.1, n =145) or a high (1.5, n =187) polyunsaturated to saturated fatty acid (P/S) ratio. The diet was started immediately after delivery. The composition of fatty acids in the breast milk and sera of the mothers and in the sera of the newborns was evaluated from a random sample of 15 mother-newborn pairs on the control diet (low P/S ratio) and 19 pairs on the experimental diet. Five days after delivery the relative amounts of fatty acids, especially that of linoleate, in the sera of the mothers differed significantly depending on the diet. Differences were also observed in breast milk samples taken three, four or five days after delivery and in the sera of the newborns sampled at the age of four or five days. Nine of the 145 newborn infants (6.2 %) in the control group had to be treated with light therapy compared with 12 out of 187 (6.4 %) of the newborn infants in the experimental group (high P/S ratio). Serum bilirubin concentrations were 142.5 μmol/l (SD 65.8) and 140.7 μmol/l (SD 73.5) in the experimental and control groups, respectively, at the age of five days. It appears that the changes in the composition of serum fatty acids reached in this study had no effect on the neonatal jaundice.     

Neonatal jaundice and fatty acid composition of the maternal diet

The role of serum fatty acid composition in neonatal jaundice was studied by comparing the incidence of jaundice among 332 newborn infants receiving breast milk from mothers on a diet with either a low (0.1, n = 145) or a high (1.5, n = 187) polyunsaturated to saturated fatty acid (P/S) ratio. The diet was started immediately after delivery. The composition of fatty acids in the breast milk and sera of the mothers and in the sera of the newborns was evaluated from a random sample of 15 mother-newborn pairs on the control diet (low P/S ratio) and 19 pairs on the experimental diet. Five days after delivery the relative amounts of fatty acids, especially that of linoleate, in the sera of the mothers differed significantly depending on the diet. Differences were also observed in breast milk samples taken three, four or five days after delivery and in the sera of the newborns sampled at the age of four or five days. Nine of the 145 newborn infants (6.2%) in the control group had to be treated with light therapy compared with 12 out of 187 (6.4%) of the newborn infants in the experimental group (high P/S ratio). Serum bilirubin concentrations were 142.5 mumol/l (SD 65.8) and 140.7 mumol/l (SD 73.5) in the experimental and control groups, respectively, at the age of five days. It appears that the changes in the composition of serum fatty acids reached in this study had no effect on the neonatal jaundice.     

The natural history of neonatal jaundice

The relationship between infant feeding type and the occurrence and natural history of neonatal jaundice in term newborn infants has been studied. A retrospective chart review of 124 records confirmed earlier reports indicating that jaundice is recognized more often in breast-fed than in formula-fed infants. A prospective cohort study of 140 term newborn infants was conducted using the Minolta Air-Shields transcutaneous jaundice meter. For 3 weeks, 115 white infants and 25 black infants were followed at predetermined intervals. The peak jaundice meter readings were higher and the elevated levels lasted longer in breast-fed than in formula-fed infants. Formula-fed infants' readings returned to base-line levels in eight days whereas the readings were still elevated in breast-fed infants when the study ended on the 21st day. Black infants had higher transcutaneous readings than white infants due to their deeper skin pigmentation, but otherwise they followed a course identical with that of the white babies. The distribution of jaundice in the white infants was bimodal; in approximately one fourth of the breast-fed infants, the jaundice meter readings reached levels corresponding to bilirubin values greater than 13 mg/dL whereas the remaining three fourths followed a pattern similar to that of the formula-fed infants. It can be concluded that human milk feeding is associated with more prolonged hyperbilirubinemia than formula-feeding in normal term infants.