Sialyloligosaccharides in human and bovine milk and in infant formulas: variations with the progression of lactation.

Several lines of research support a role for human milk oligosaccharides in the defense of breast-fed infants against pathogens. Some ofthese oligosaccharides contain at least one moiety of sialic acid and are, thus, termed sialyloligosaccharides. These constitute a significant component (>1 g/L) of human milk. It is well established that milk composition varies among species, and previous reports have indicated that one ofthe differences between human and bovine milk is precisely their contents of sialyloligosaccharides. Because most infant formulas are manufactured with bovine milk components, it follows that formula-fed and breast-fed infants ingest dissimilar quantities of these carbohydrate structures. To ascertain these differences and their impact along lactation, the contents of oligosaccharide-bound sialic acids and major sialyloligosaccharides in samples of human and bovine milk (obtained at different lactation stages) were determined. In addition, infant formulas were assayed for their sialyloligosaccharide contents. Seven sialyloligosaccharides were identified in human milk; namely, 3'-sialyl-3-fucosyllactose and sialyllacto-N-tetraoses (a and b+c), the predominant structures at all lactation stages. Five sialyloligosaccharides were identified in bovine milk, of which 6'-sialyllactosamine and 3'-sialyllactose were the most abundant. In addition, sialyloligosaccharides in human and bovine milk decreased along lactation, and infant formulas did not contain significant amounts of sialyloligosaccharides. The results point to the general conclusion that regarding both qualitative and quantitative aspects, milk from humans and cows and infant formulas have different oligosaccharide contents. In this sense, bottle-fed infants are subject to reduced sialyloligosaccharide intake as compared to breast-fed infants.

Distribution of bovine milk sialoglycoconjugates during lactation.

The sialoglycoconjugate content of human milk has been extensively studied. However, little attention has been paid to the changes occurring in these compounds in bovine milk during lactation. Since sialoglycoconjugates are very abundant in milk from the early stages of lactation, they have been suggested to be important for the nutrition of the newborn during the first months of life. The distribution of sialoglycoconjugates (expressed as glycoconjugate-bound sialic acid) from four different stages of lactation (colostrum, transitional, mature, and late-lactation milks) was investigated in four Spanish-Brown cows. All the fractions studied (total sialic acids, glycoproteins, oligosaccharides, casein, and gangliosides) showed a similar trend. We found the highest values in the colostrum, these decreasing in transitional and mature milks and increasing again in late-lactation milk. We also found a selective change in the relative contents of glycoprotein- and oligosaccharide-bound sialic acids. In mature milk, the latter increased up to 80% (59% in colostrum) and the former decreased to 3.9% (35.3% in colostrum). It would appear that the decrease in oligosaccharide-bound sialic acid is compensated by the increase in glycoprotein-bound sialic acid. From these results, it is deduced that newborn infants or calves fed with infant formulas or milk replacers, respectively, should be supplemented with sialoglycoconjugates to approximate the composition of human and cow milk as far as is practicable.

Developmental changes in UDP-N-acetylglucosamine 2-epimerase activity of rat and guinea-pig liver.

The activity of UDP-N-acetylglucosamine 2-epimerase was determined in the liver of rats and guinea-pigs of different ages. The activity of this enzyme in rats was low at birth, increased to a maximum value on day 15, and fell gradually until day 30. Thereafter, it increased up to the 60th day. The activity profile of the enzyme from guinea-pig liver was very similar. However, guinea-pig activity was 2-5 times lower than in rats. Both rats and guinea-pigs displayed similar liver sialic acid contents which increased from birth to 2 months of age. Rats also showed a N-glycolylneuraminic acid content that decreased from birth to 2 months. From these results we can inferred that postnatal UDP-N-acetylglucosamine 2-epimerase activity seems to be correlated with age and the developmental states of rats and guinea-pigs.

Changes in ganglioside and sialic acid contents of goat milk during lactation.

The ganglioside content of goat milk has been determined from d 1 after parturition to d 60 of lactation. Marked changes occurred in milk over the course of lactation; the highest ganglioside content occurred in d-1 colostrum and then decreased to the end of the period studied. At least seven different ganglioside species were detected; three gangliosides containing sialyllactosylceramide accounted for 66 to 92% of the total lipid-bound sialic acid; this result reflected a very simple core structure of goat milk gangliosides. The most abundant ganglioside, II3(N-acetylneuraminic acid)2-lactosylceramide, was about 35 to 56%. The sialic acid content exhibited a trend similar to that of gangliosides; during early lactation sialic acid content was higher than in mature milk. Fat, protein, and total solids were high at initiation of lactation and decreased thereafter. However, lactose content remained almost unchanged during the period studied.

Alterations induced by fascioliasis and cirrhosis on the biliary excretion of cefmetazole in Wistar rats.

1. Alterations induced by fascioliasis and cirrhosis on the biliary excretion of cefmetazole have been studied in Wistar rats. 2. Both infestation with Fasciola hepatica and experimental cirrhosis originated a significant decrease in the biliary excretion and in bile flow increase induced by the drug. 3. Administration of the beta-lactam antibiotic induced a lower degree of uncoupling of biliary lipid secretion in the cirrhotic and fasciolotic animals, but the effect was evident in all experimental groups.

Gangliosides in bovine milk. Changes in content and distribution of individual ganglioside levels during lactation.

Bovine milk undergoes changes in its ganglioside contents during the different stages of lactation. These contents are higher in colostrum (7.5 mg of lipid-bound NeuAc/kg) than in transitional (2.3 mg) or mature (1.4 mg) milk. The sialic acid content of milk follows a similar profile to that of gangliosides with the highest content during the first few days post partum followed by a gradual decrease towards the end of the period studied. When the individual distribution of gangliosides was examined throughout the course of lactation, several changes were also found. GD3 is the major ganglioside (about 60-70%) found; its content decreases from the first to the fifth day, increasing towards the end of the period considered. GM3, GD3 and GT3, sialyllactosylceramide-containing gangliosides account for 80-90% of the total lipid-bound NeuAc content. The most striking change in the ganglioside pattern was the gradual increase in G3.

Hepatic transport of bilirubin in rats with streptozotocin-induced diabetes.

This study was undertaken to evaluate the effects of streptozotocin-induced diabetes on the hepatic transport of bilirubin in male Wistar rats. Rats were pretreated with streptozotocin (60 mg/kg i.p.) to induce uncontrolled diabetes. Six days later endogenous biliary excretion and plasma bilirubin concentration were significantly enhanced compared to control animals (+36% and +46%, respectively), while the blood levels of free hemoglobin remained unchanged. Following a bilirubin load, the maximal biliary excretion of the pigment (Tm) in diabetic animals was significantly enhanced compared to control animals (+49%). Liver and plasma bilirubin concentrations at the end of bilirubin administration were significantly reduced (-28% and -30%, respectively). Bilirubin UDP-glucuronosyltransferase activity and UDP-glucose concentration in liver were significantly enhanced (+31% and +81%, respectively), as was the biliary excretion of unconjugated bilirubin (+37%) and bilirubin mono- (+38%) and diconjugates (+53%). When streptozotocin-diabetic rats were treated with insulin, the parameters of bilirubin transport and metabolism were significantly reduced compared to diabetic animals receiving no hormone replacement. In summary, our data indicate that in short-term streptozotocin-diabetic rats there is increased bilirubin production as well as enhanced hepatic conjugation and subsequent biliary excretion of the pigment. These effects appear to be a direct consequence of diabetes.

Changes in hepatic cytosolic glutathione S-transferase enzymes induced by clotrimazole treatment in rats.

1. The influence of the antifungal agent clotrimazole on cytosolic glutathione S-transferase activities was studied in male Wistar rats. 2. Animals received clotrimazole by gastric lavage for 3 days (75 mg/kg per day). Hepatic glutathione S-transferase activity was determined with five different substrates: 1-chloro-2,4-dinitrobenzene (CDNB), 1,2-dichloro-4-nitrobenzene (DCNB), p-nitro-benzyl chloride (PNBC), ethacrynic acid (EA) and trans-4-phenyl-3-buten-2-one (TPBO). 3. The largest increases in glutathione S-transferase activity were found with CDNB, DCNB and PNBC (+61%, +50% and +50%, respectively, when expressed per mg of cytosolic protein). Enzyme activity toward EA was induced to a lower extent (+33%). Changes in the formation of the conjugate of TPBO were relatively small (+22%). 4. These data indicate a differential induction of glutathione S-transferase isoenzymes and suggest that clotrimazole is a phenobarbital-type inducer of enzyme activity.

Sex-related differences in the hepatobiliary transport of phenolsulfonphthalein in the rat.

Sex-related differences in the hepatobiliary transport of phenolsulfonphthalein (PSP) were investigated in male and female Wistar rats. Maximal biliary excretion of unconjugated PSP was significantly higher in females while the excretion of the conjugated dye and liver UDP-glucuronosyltransferase activity toward PSP were higher in male animals. Orchidectomy decreased enzyme activity and excretion of the conjugate, whereas ovariectomy produced the opposite effect. Both in gonadectomized males and females maximal biliary excretion of the unconjugated dye was significantly reduced. Testosterone treatment increased the excretion of both conjugated and unconjugated PSP and transferase activity in orchidectomized males. Combined treatment of gonadectomized females with estradiol plus progesterone led to excretions of both conjugated and unconjugated PSP and UDP-glucoronosyltransferase activities similar to those found in control rats. These data indicate the existence of sex-related differences in the conjugation and biliary excretion of PSP in the rat and its modulation by sex hormones.

Postnatal development of the hepatobiliary transport of phenolsulfonphthalein in rats.

1. The postnatal development of the biliary excretion of phenolsulfonphthalein (PSP) was studied in male Wistar rats. 2. Following i.v. injection of PSP at 200 mumol/kg body wt, a maximal biliary excretion of 175 +/- 10 nmol/min/100 g body wt and 32 +/- 5 nmol/min/100 g body wt was reached for unconjugated and conjugated PSP, respectively, in the adult group. 3. The maximal biliary excretion of conjugated PSP was significantly lower in the 20-, 30- and 40-day-old groups as compared to the adults. The excretion of unconjugated dye was also significantly lower in 20- and 30-day-old rats. 4. The postnatal development of PSP excretion was unrelated to changes in the activity of UDP-glucuronosyltransferase. The importance of other factors is also discussed.