Effects of term infant formulas containing high sn-2 palmitate with and without oligofructose on stool composition, stool characteristics, and bifidogenicity.

OBJECTIVES: Levels of stool fatty acid soaps and beneficial bacteria differ between formula-fed and breast-fed infants; addition of specific formula ingredients may reduce these differences. This study evaluated the effects of a term infant formula containing high sn-2 palmitate term infant formula (sn-2) or an identical formula supplemented with oligofructose (OF) at 2 concentrations (sn-2+3 g/L OF, sn-2+5 g/L OF) on stool composition, stool characteristics, and fecal bifidobacteria. METHODS: Healthy, term formula-fed infants 7 to 14 days old (n = 300) were randomized in a double-blind manner to receive standard formula (control), sn-2, sn-2+3 g/L OF, or sn-2+5 g/L OF for 8 weeks. Human milk (HM)-fed infants (n = 75) were studied in parallel. Stool samples were collected from all subjects at week 8 for fatty acid soaps and mineral content, and from a subset at baseline and week 8 for bifidobacteria. Stool characteristics were assessed via 3-day diary. RESULTS: The sn-2 group had 46% less stool soap palmitate (P < 0.001) and softer stools than control (20% more mushy soft stools, P = 0.026; 50% fewer formed stools, P = 0.003). Addition of OF resulted in even fewer formed stools versus control (65% fewer for sn-2+3 g/L OF, 79% fewer for sn-2+5 g/L OF), with 5 g/L OF more closely resembling that of HM-fed infants. Both sn-2 (P < 0.05) and sn-2 with OF groups (P < 0.01) had significantly higher fecal bifidobacteria concentrations than control at week 8, not differing from HM-fed infants. CONCLUSIONS: High sn-2-palmitate formulas led to reduced stool soaps, softer stools, and increased bifidobacteria, whereas addition of OF further improved stool consistency. Those modifications brought outcomes in formula-fed infants closer to that in HM-fed infants.

Nutritional influences on visual development and function.

Experiments conducted on many different species reveal a fundamental paradox about the vertebrate eye; it is damaged by its own operation. This vulnerability stems from the need to respond to visible light, often actinic, but also from the intrinsic metabolic and structural state of the eye's internal structures. Photoreceptor outer segments, for instance, have high concentrations of diet-derived long-chain polyunsaturated fatty acids and these membrane lipids are highly prone to peroxidation due to the high oxygen tension of the outer retina. Such a high diathesis for damage would be catastrophic if it were not balanced by an equally impressive system for responding to such stressors. The retina (and to a lesser extent the crystalline lens), for instance, is especially rich in dietary antioxidants such as vitamin E, vitamin C and the macular carotenoids (lutein and zeaxanthin) putatively to retard light-induced oxidative damage. The nutrients that support both essential function (e.g., retinal, the vitamin form of vitamin A, in photopigment) and protection operate in a highly integrated manner. For instance, Vitamin E is a lipophillic chain-breaking anti-oxidant (protecting DHA-rich outer segment membranes) that regenerates itself through reaction with vitamin C (a primary anti-oxidant against aqueous radicals) and is spatially distributed in complement with the carotenoids lutein and zeaxanthin. Nor are these interactions relegated to simply providing protection and the basic elements needed for transduction. Macular lutein and zeaxanthin, for example, improve visual performance (e.g., reduce glare disability and discomfort, speed photostress recovery, and enhance chromatic contrast) through purely optical means (by absorbing short-wave light anterior to the foveal cones). The vulnerability of the eye to exogenous insult, and the sensitivity of the eye to dietary components, is not static: infants have more vulnerable retinas due to clearer lenses and higher metabolic activity; the elderly are more vulnerable due to such factors as increased inflammatory stress and a higher content of photosensitizers (such as lipofuscin) creating cascading oxidative effects. Hence, optimal dietary prophylaxis changes as the eye ages. The eye, perhaps more than most other biological structures, has evolved an exquisite and shifting sensitivity to dietary intake throughout the lifespan, not just for its basic operation (e.g., Vitamin A for transduction), but also for its very preservation.

Toxicology and safety of DHA.

Docosahexaenoic acid (DHA) is a long-chain polyunsaturated fatty acid with activities in both infants and adults. The objective of the current work was to evaluate the published literature concerning the toxicological assessment of DHA-rich oils in animals and the safety profile of DHA consumption in humans. Structured literature searches concerning DHA toxicology and DHA effects on platelet function, lipid levels, oxidative potential, glycemic control, and immune function were conducted. The toxicological profile of DHA derived from single-cell organisms demonstrates that these oils are safe in rats (up to a consumption level of 3290 mg/kg body weight/d) in 90-d toxicology evaluations, as well as in reproductive and developmental toxicology studies. The maximum DHA level in human breast milk exceeds 1% of total fatty acids in high-fish-consuming populations. Consumption of DHA-rich human milk as sole source of nutrition provides approximately 315 mg/d in infants 1-6 months of age, and appears to be a safe level of intake. DHA supplementation studies in adults have employed doses ranging from less than 1 to 7.5 g/d, and have not resulted in any consistent adverse responses in platelet function, lipid levels, in vivo oxidation parameters, glycemic control, or immune function. In conclusion, DHA consumption does not result in consistent adverse events in infants or adults. Safe intake levels may be modeled on DHA intake from human milk in infants, and may be at least as high as the upper doses studied in adults.

Safety evaluation of oligofructose: 13 week rat study and in vitro mutagenicity.

Oligofructose (OF), comprised of fructose oligomers with a terminal glucose unit, is a family of oligosaccharides derived from the hydrolysis of inulin. Consumption of OF in animals and humans increases colonic bifidobacteria levels. The present study evaluates the safety of OF in both a 13 week rat feeding study and using in vitro mutagenicity tests. Fecal bifidobacteria levels were also determined by in situ hybridization to assess a biological function of OF. Rats received either a control diet or diets containing one of four doses of OF. Total, HDL, and LDL-cholesterol levels were significantly lower at several time points during the study in groups receiving OF compared to controls with the largest effects occurring in the high dose male animals. Weight gain in the male high dose group was significantly lower at early time points compared to controls but not significantly different at the end of study. As expected, cecal weights increased in a dose-related manner and fecal bifidobacteria levels also demonstrated a dose-related increase. There were no consistent differences in gross pathology or histopathology related to dietary OF. OF did not induce a positive response in the Ames test or chromosomal aberration test with CHO cells. These results demonstrate no adverse effects of OF.

Human milk fatty acid composition from nine countries varies most in DHA.

Many published studies of breast milk FA composition are limited to populations from one or two countries. We aimed to examine the degree to which FA compositions vary across a number of diverse populations. Because diet and maternal adipose stores influence breast milk FA composition, differences in FA composition between groups most likely reflect habitual dietary differences. Approximately 50 breast milk samples (full breast expression) were collected from women in Australia, Canada, Chile, China, Japan, Mexico, Philippines, the United Kingdom, and the United States. The proportion of saturated FA was relatively constant among countries, with the exception of the Philippines, where levels of lauric and myristic acids were elevated (means greater than two times the mean of most other countries). Monounsaturated FA also varied little, with the exception of low levels of oleic acid in the Philippines and high levels of erucic acid in China. Although arachidonic acid (C20:4n-6) levels were similar among all countries (means ranging from 0.36 wt % to 0.49 wt %), mean DHA (C22:6n-3) levels ranged from 0.17 to 0.99 wt %, with the highest levels in Japanese milk and the lowest levels in Canadian and U.S. samples. The results of this study demonstrate that the proportion of saturated and monounsaturated FA are relatively constant across a large number of countries, whereas the level of some of the PUFA, especially DHA, are highly variable.

Detection of a single nucleotide polymorphism in the human alpha-lactalbumin gene: implications for human milk proteins.

Variability in the protein composition of breast milk has been observed in many women and is believed to be due to natural variation of the human population. Single nucleotide polymorphisms (SNPs) are present throughout the entire human genome, but the impact of this variation on human milk composition and biological activity and infant nutrition and health is unclear. The goals of this study were to characterize a variant of human alpha-lactalbumin observed in milk from a Filipino population by determining the location of the polymorphism in the amino acid and genomic sequences of alpha-lactalbumin. Milk and blood samples were collected from 20 Filipino women, and milk samples were collected from an additional 450 women from nine different countries. alpha-Lactalbumin concentration was measured by high-performance liquid chromatography (HPLC), and milk samples containing the variant form of the protein were identified with both HPLC and mass spectrometry (MS). The molecular weight of the variant form was measured by MS, and the location of the polymorphism was narrowed down by protein reduction, alkylation and trypsin digestion. Genomic DNA was isolated from whole blood, and the polymorphism location and subject genotype were determined by amplifying the entire coding sequence of human alpha-lactalbumin by PCR, followed by DNA sequencing. A variant form of alpha-lactalbumin was observed in HPLC chromatograms, and the difference in molecular weight was determined by MS (wild type=14,070 Da, variant=14,056 Da). Protein reduction and digestion narrowed the polymorphism between the 33rd and 77th amino acid of the protein. The genetic polymorphism was identified as adenine to guanine, which translates to a substitution from isoleucine to valine at amino acid 46. The frequency of variation was higher in milk from China, Japan and Philippines, which suggests that this polymorphism is most prevalent in Asia. There are SNPs in the genome for human milk proteins and their implications for protein bioactivity and infant nutrition need to be considered.

A multinational study of alpha-lactalbumin concentrations in human milk.

Alpha-lactalbumin, a 14-kD protein, plays a central biochemical role in the mammary gland as the regulatory subunit of lactose synthase, and also plays a nutritional role for the rapidly growing neonate as the protein in highest concentration in human milk. The current study was undertaken to better characterize alpha-lactalbumin concentrations in human milk from a variety of countries. Mature human milk (lactation duration > or =1 month) was collected from at least 50 women from nine different countries on five continents. Alpha-lactalbumin concentration was determined by HPLC. The mean +/- SD for 452 samples was 2.44 +/- 0.64 g/L. The mean value of the samples from the United States was significantly higher than that from any other country, and the mean in Mexico was significantly lower than that from every country except China and Canada. Alpha-lactalbumin concentration decreased with increasing duration of lactation and was positively correlated with total nitrogen. On average, alpha-lactalbumin contributed 16% of the total nitrogen content of human milk and consequently an important part of the amino acid content.

Growth and safety in term infants fed reduced-protein formula with added bovine alpha-lactalbumin.

OBJECTIVE: To assess growth, tolerance, and biochemical measures of protein status in term infants fed an experimental formula with reduced total protein concentration and enriched in bovine alpha-lactalbumin prepared from an alpha-lactalbumin dominant bovine whey. METHODS: Healthy, term, exclusively formula-fed infants < or =14 days postnatal age, between 10th and 90th percentiles in weight and length for age were studied in this randomized, masked, multicenter study. Infants received ad libitum feedings of either experimental or control formula for 12 weeks. Adverse events and acceptability of formulas were assessed every 2 weeks. Weight, length, and head circumference were measured at baseline and every 4 weeks. Serum creatinine, albumin, and blood urea nitrogen were assessed at baseline and study completion. Anthropometric measures were compared to Centers for Disease Control reference ranges using Z scores. RESULTS: One hundred ninety-three infants were enrolled. One hundred thirty-four completed the protocol. Seventy-two received experimental formula, and 62 received control formula. The mean baseline body weight was higher in infants fed experimental formula (P = 0.042), so baseline weight was used as a covariate in statistical analyses. There were no differences between groups in gains in weight, length, or head circumference during the study. Mean serum albumin and blood urea nitrogen (BUN) levels were similar at study initiation. At study completion, serum albumin levels were the same in both groups, whereas BUN was significantly higher in infants fed control formula (P = 0.0016). More infants fed control (n = 20) than experimental (n = 15) formula discontinued the study because of adverse events. There were no differences in the adverse event profiles of the groups completing the study. Most of the adverse events were mild and resolved without treatment or sequelae. Acceptability and tolerance of the experimental formula was greater than the control formula, except at 2 weeks. Unacceptable ratings ranged from 0% to 10% (mean, 4.1%) in the experimental formula group and from 1.6% to 14.1% (mean, 7.0%) in the control formula group. CONCLUSIONS: Growth and serum albumin were comparable in infants fed experimental and control formulas for the first 12 weeks of life, suggesting adequate protein nutrition from the alpha-lactalbumin-rich formula, despite its lower total protein content. Growth and adverse events data support the safety of the experimental formula. Fewer discontinuations and unsatisfactory ratings among infants fed the experimental formula suggest that it is better tolerated than control formula.

Nutritional and physiologic significance of alpha-lactalbumin in infants.

alpha-Lactalbumin is the major protein in breast milk (20-25% of total protein) and has been described to have several physiologic functions in the neonatal period. In the mammary gland, it participates in lactose synthesis, thereby creating an osmotic "drag" to facilitate milk production and secretion. alpha-Lactalbumin binds divalent cations (Ca, Zn) and may facilitate the absorption of essential minerals, and it provides a well-balanced supply of essential amino acids to the growing infant. During its digestion, peptides appear to be transiently formed that have antibacterial and immunostimulatory properties, thereby possibly aiding in the protection against infection. A novel folding variant ("molten globule state") of multimeric alpha-lactalbumin has recently been discovered that has anti-infective activity and enhances apoptosis, thus possibly affecting mucosal cell turnover and proliferation. Cow milk also contains alpha-lactalbumin, albeit less than human milk (2-5% of total protein in bovine milk), and protein fractions enriched with alpha-lactalbumin may now be added to infant formula to provide some of the benefits of human alpha-lactalbumin.

Infant formulas with increased concentrations of alpha-lactalbumin.

Human and bovine milk differ substantially in the ratio of whey to casein protein (approximately 60:40 in human milk and approximately 20:80 in bovine milk) and in the proportions of specific proteins. Although current infant formulas closely mimic the ratio of total whey to casein inhuman milk, the concentration of a-lactalbumin (the dominant protein in human milk) is relatively low in formula, whereas beta-lactoglobulin, a protein not found in human milk, is the most dominant whey protein in formula. Because of the differences in the protein profiles of human milk and infant formula, amino acid profiles also differ. To meet all essential amino acid requirements of infants, formula concentrations of protein must be higher than those in human milk. Recently, whey sources with elevated concentrations of alpha-lactalbumin have become available, which permitted the development of formulas with increased concentrations of this protein and decreased concentrations of beta-lactoglobulin. alpha-Lactalbumin is rich in tryptophan, which is typically the limiting amino acid in formula, and as a result, formulas have been developed with lower protein but higher tryptophan concentrations. This type of formula may offer a number of advantages to the neonate, which include producing plasma tryptophan concentrations equal to those found in breastfed infants and obviating the need for the body to dispose of excess nitrogen loads.