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Speciation of zinc in low molecular weight proteins of breast milk and infant formulas by size exclusion chromatography/flame atomic absorption spectroscopy.

Size exclusion chromatography (SEC) and flame atomic absorption spectroscopy (FAAS) were used for the separation of metal-containing species in milk whey. After milk ultracentrifugation, the sample was injected into a TSK-Gel G2000 glass column and eluted with 0.2M NH4NO3-NH3, pH 6.7. Low molecular weight proteins were fractionated, and the fractions were characterized by molecular weight. Zinc distributions were obtained by FAAS using a high performance nebulizer. The method was very sensitive (limit of detection = 2.6 x 10(-3) microg/mL; limit of quantitation = 8.9 x 10(-3) microg/mL) and precise (RSDs < or =10%). This method was applied to the determination of Zn in binding compounds in breast milk whey and in commercial cow's milk-based formulas. Distribution patterns were different. The presence of Zn in most fractions in breast milk was most significant, whereas in infant formulas Zn was detected only in fractions of molecular weight <5 kDa and in the highest molecular weight peak.

Calibration↗

The interaction of infant formula with macrophages: effect on phagocytic activity, relationship to expression of class II MHC antigen and survival of orally administered macrophages in the neonatal gut.

The effect of infant formula on human peritoneal and breast milk macrophages has been investigated. The ability of peritoneal macrophages to subsequently ingest and degrade immune complexes was slightly impaired, but breast milk cells were not affected. However, the cells were found to have bound antigenically intact casein and beta-lactoglobulin, although little, if any, alpha-lactalbumin was bound. Furthermore, a positive correlation was found between binding of these proteins and expression of HLA-DR antigen. Labelled macrophages fed to newborn mice survived for at least 4 hr in the gastrointestinal tract and, in some cases, localized in the mucosal tissue. In one case a labelled cell was found in the spleen. These findings indicate that breast milk macrophages may be able to perform immunological functions in the gut, and suggest that binding of cows' milk proteins by macrophages may constitute a first step in the sensitization of the neonate to cow's milk proteins. Human milk macrophages may also play a protective role by acting as antigen-presenting cells in the local immune response of the gut.

Animals↗

Amino acid ratings of different forms of infant formulas based on varying degrees of processing.

Amino acid profiles, protein digestibility and/or amino acid bioavailability for the various forms (powder, liquid concentrate, ready-to-use, etc.) of infant formulas (involving varying degrees of heat processing during preparation) have been determined. Amino acid scores (based on the single most limiting amino acid) were calculated by comparing the essential amino acid data with that of human milk. Amino acid scores were multiplied by total protein (g/100 kcal) to obtain amino acid ratings, which take into account both quality and quantity of protein. Amino acid scores for milk- and soy-based formulas ranged from 49 to 90 and 59 to 81%, respectively, due to deficiencies in methionine plus cystine and/or tryptophan. The deficiency in the limiting amino acids was more marked in liquid concentrate than powder prepared by the same manufacturer. Because of significantly higher total protein contents (g/100 kcal) of soy- (2.65-3.68) and milk-based (2.20-2.95) formulas compared to human milk (1.5), the relative amino acid ratings (human milk = 100) of all formulas except two milk-based liquid concentrates and one ready-to-feed (with values of 77-87%) were greater than 100%. When corrected for protein digestibility, the relative amino acid ratings for all four liquid concentrates were less than 100%. Lower levels of digestible protein and bioavailable amino acids in liquid concentrate compared with powder (prepared by the same manufacturer) would suggest that inferior protein quality of liquid concentrates may be due to more severe heat treatment involved in their preparation.

Amino Acids↗

A novel infant formula milk with added long-chain polyunsaturated fatty acids from single-cell sources: a study of growth, satisfaction and health.

OBJECTIVE: To evaluate the growth, feeding and health of babies fed a novel infant formula milk with added long chain polyunsaturated fatty acids (LCPs) produced from single-cell sources, at concentrations similar to those found in mature breast milk. DESIGN: Randomized double-blind control trial. SUBJECTS: One-hundred and forty healthy, full-term infants of birth weight 2.5-4.5 kg born at the Maternity Department, East Glamorgan General Hospital, Wales, whose mothers had already decided to bottle feed. INTERVENTIONS: Subjects were randomized to two groups; one (control group) to receive a standard formula milk and the other to receive the trial milk with added LCPs. Milks were supplied in a double-blind fashion and given for the first 12 weeks of life. Anthropometric measurements were made at recruitment, 3 months, 6 months and 1 y. Feeding diaries were completed at 6 weeks and 3 months, and a parental record was kept of any ill health suffered by the subjects during the first year of life. RESULTS: Of 140 infants recruited, 31 did not complete the protocol. Small but statistically significant differences were found only in the subscapular skinfold thickness at 6 weeks and 3 months, that in the trial group being slightly higher, but unlikely to be of any clinical importance. No differences were found in the feeding patterns of the infants in the two groups. Stool patterns were similar, as were the frequencies of illness and allergy. CONCLUSIONS: This study supports the view that LCPs from single-cell sources do not have detrimental effects on the growth, feeding and general health of infants.

Anthropometry↗

Rearing of gnotobiotic rodents by feeding a commercially-prepared infant formula.

A simple, inexpensive method for the rearing of germfree rats was developed. Germfree rats were fed from age 10 days to 40 weeks a diet consisting of commercially prepared Similac infant formula, water, and corn oil. Weight gains and gross appearances were comparable to littermate controls. Antibodies to bacterial teichoic acid were present in all controls while absent in rats fed the Similac diet.

Animal Nutritional Physiological Phenomena↗

Efficacy of a pre-thickened infant formula: a multicenter, double-blind, randomized, placebo-controlled parallel group trial in 104 infants with symptomatic gastroesophageal reflux.

To evaluate a pre-thickened formula (Enfamil AR) for regurgitant gastroesophageal reflux, 104 infants were enrolled in a 5-week, multicenter, double-blind, randomized, placebo-controlled parallel group trial. The Enfamil AR group showed greater symptom reduction by the end of the first week: percent feedings with any regurgitation (p = 0.045), total regurgitation volume score (p = 0.035), and percent feedings with choke-gag-cough (p = 0.004). The most symptomatic infants at baseline had a reduction in trouble sleeping significantly with Enfamil AR by the end of the study (p = 0.030). This formula flows through a standard nipple, reduces regurgitation and choking-gagging-coughing within a week, and improves sleep in the most symptomatic babies by 5 weeks, without causing constipation.

Double-Blind Method↗

[Life-threatening hypernatremia caused by faulty preparation of fully adapted infant formula].

Two male newborns developed severe life threatening hypernatremia with serum sodium levels of 181 and 196 meq/l respectively. Both children were fed a two-to fivefold concentrated powdered infant formula since birth. Shortly before admission diarrhea developed in both patients. The children were admitted during the third week and fourth week respectively, of their lifes with clinical signs of severe hyperosmolar dehydration. In both, treatment of shock was instituted and was followed by rehydration therapy. Great care was taken to lower serum sodium by not more than 15 meq/l/day in order to prevent cerebral edema. Inspite of these measures, the first patient developed cerebral seizures 5 hours after initiation of fluid replacement. The patient was intubated, and measures aimed at prevention of cerebral edema were started (hyperventilation). The eventual outcome was satisfactory, on follow up the patient showed no signs of persistent cerebral damage. In the second patient the same management was started from the beginning and no cerebral problems occurred. Restoration of stable body functions with a normal electrolyte status was achieved within five days.

Brain↗

Antigen-reduced infant formulas versus human milk: growth and metabolic parameters in the first 6 months of life.

OBJECTIVE: To compare growth and metabolic parameters of infants fed special formulas for atopy prevention to breast-fed infants. METHODS: Eighty-two infants with family history of IgE-mediated diseases completed a 6-month nutritional follow-up within a clinical trial to assess the effectiveness of the dietary intervention in preventing atopy. Infants were subdivided according to the type of milk feeding from the end of the first month of life up to sixth month: human milk (HM) group (29 infants); soy formula (SF) group (15 infants); whey-based low-degree hydrolysate (WHy) group (15 infants); casein-based high-degree hydrolysate (CHy) group (13 infants); soy plus collagen-based high-degree hydrolysate group (10 infants). Anthropometric indices were determined at 0, 3 and 6 months of life and a fasting blood sample for hematochemical parameters was obtained at 5 months of life before beginning the introduction of solid foods. RESULTS: Lower values of body mass index and higher blood urea nitrogen were displayed by the CHy group at 3 months. Plasma aminoacidograms showed higher essential amino acids (AA)/total AA in SF, WHy and CHy groups but lower branched chain AA/essential AA in all formula-fed infants in comparison with the HM group. CONCLUSIONS: Special formulas (in particular high-degree protein hydrolysates) should be carefully evaluated when used for prolonged periods of time in healthy infants at familial risk of atopy.

Amino Acids↗

Multiple analysis of a foodborne outbreak caused by infant formula contaminated by an atypical Salmonella virchow strain.

An outbreak of food poisoning involving most autonomous Spanish communities was detected in the first half of 1994. The causative food was infant formula milk contaminated by lactose-fermenting Salmonella virchow. It was not possible to isolate the causative strain from the manufacturer's facilities. During the same period of time, there was a significant increase in lactose-non-fermenting Salmonella virchow strains compared with the same period in previous years. Simultaneously, lactose-non-fermenting strains were recovered from clinical samples from children and from some milk samples that were involved in the outbreak. Therefore, it was speculated that the outbreak might be more extensive than initially thought. The following epidemiological markers were used for typing the Salmonella virchow strains involved in the outbreak: (i) phage typing: (ii) ribotyping, using a set of 20 different endonucleases: and (iii) pulsed-field gel electrophoresis, using three different endonucleases. The most useful markers for this serotype were phage typing and pulsed-field gel electrophoresis, since ribotyping was not able to distinguish all strains tested. The results obtained revealed that the outbreak was caused by at least two strains: one presenting phage type 4-4a and pulsed-field patterns A1 or A2 and L+ or L-, and another presenting phage type 2 and pulsed-field patterns A1 or A2 and L+ or L-. The results indicate that the outbreak was more extensive than initially thought and that the Virchow serotype is very clonal in Spain.

Animals↗

Problems associated with the establishment of maximum nutrient limits in infant formula.

Because each increase in personal security is accompanied by a small but measurable decrease in individual liberty, congress and the American people have been reluctant to provide to the regulatory agencies authority beyond that which they believe is the minimum necessary to avoid a crisis. For government to act in any area involving nutrient fortification of foods requires an acceptable database demonstrating a health hazard. The requirements of current law demand that new formulas be investigated by appropriate clinical studies but do not grant the FDA clear authority to establish maximum limits for nutrients in infant formulas. Except when an unquestionable hazard can be demonstrated, there are important barriers to governmental action in this area. Without a clearly defined, scientifically acceptable database, without consensus in the scientific and professional community, and without public support for action to be taken in this area, the establishment of maximum limits for many nutrients will be extremely difficult.

Female↗

Saturated fatty acid chain length and positional distribution in infant formula: effects on growth and plasma lipids and ketones in piglets.

Human milk contains a large proportion of palmitic acid (16:0) with > 70% esterified to the center sn-2 position of the milk triglyceride. Infant formulas often use 8:0 + 10:0 [medium-chain triglyceride (MCT)] or 12:0 + 14:0 (coconut oil) as the saturated fat. The effect of formula saturated fatty acid composition; 8:0 + 10:0, 12:0 + 14:0, or 16:0 from palm oil or synthesized triglyceride containing predominantly sn-2 16:0 on plasma lipids and fatty acids was studied in piglets. Although the formulas contained similar 18:1 and 18:2n-6, plasma lipid percentages of 18:1 and 18:2n-6 were higher in piglets fed the formula with MCT or coconut oil rather than the formulas with 16:0, or sow milk. The sn-2 16:0 of the synthesized triglyceride had unique properties. Specifically, piglets fed synthesized triglyceride had significantly higher cholesteryl ester 16:0 identical to that in piglets fed sow milk and higher plasma total and high-density-lipoprotein cholesterol than piglets fed the other formulas.

Animals↗

Infant formula samples and breast feeding among Philippine urban poor.

An experiment was performed on the maternity wards of three public hospitals in Cebu City, Philippines to determine whether the distribution of free samples of infant formula reduced the likelihood that mothers would breast feed or caused mothers to terminate nursing early. Samples were given or withheld alternately for 2 week intervals to mothers as they left maternity wards. They were followed for 8 months in the first experiment (N = 273) and for 2 months in a replication (N = 284). We found that there were no statistically significant differences between those who received samples and those who did not in initiation or maintenance of breast feeding. Mothers in both groups frequently turned to mixed schedules, but these varied from day to day depending on money to buy other forms of milk, or on the mother's health, or her plan to be away from the baby for one or more feeding periods. After the baby reached an age of 2-3 months, mothers, with few exceptions, used diluted sweetened condensed milk as a supplement and/or substitute for their own milk. It was found that, while mothers recognize the nutritional, economic and health benefits of breast feeding, they may terminate early on the basis of folk beliefs. Receiving formula samples, however, had no measured effect on their breast feeding practices.

Adult↗

Leptin levels in preterm human breast milk and infant formula.

OBJECTIVE: Leptin, a hormone present in breast milk, is involved in energy regulation and metabolism. The purpose of this investigation was to determine whether leptin is present in either preterm breast milk (PBM) or preterm formula (PF). The effects of delivery methods and pasteurization on leptin levels also were evaluated. METHODS: PBM samples were obtained from 29 mothers who delivered infants at between 23 and 34 weeks' gestation. Leptin levels were measured in PBM and PF with the use of a radioimmunoassay specific for human and bovine leptin, respectively. Milk samples were pasteurized by fast- and slow-heating methods. PBM and PF spiked with human leptin were delivered through catheters by bolus and continuous administration to determine the effects of delivery method on recoverable leptin levels. RESULTS: Median PBM leptin concentration was 5.28 ng/mL (intraquartile range: 24.79). Birth gestational age, birth weight, and gender of the infant did not significantly influence PBM leptin levels. Neither bolus nor continuous feeding practices affected leptin levels in PBM or spiked PF. However, pasteurization significantly reduced the amount of detectable leptin in PBM. CONCLUSIONS: PBM leptin levels were highly variable and similar to levels reported for term breast milk. There was no effect of postnatal age on PBM leptin concentrations. Sterilization decreased detectable leptin levels, whereas feeding practices had no adverse effect on the quantity of leptin delivered. Although no infant formula contained leptin, leptin could be added to formula and delivered through various feeding methods without loss.

Blotting, Western↗

Elevated intakes of zinc in infant formulas don not interfere with iron absorption in premature infants.

BACKGROUND: Zinc and iron may share common pathways for absorption and compete for uptake into mucosal cells. We determined whether elevated ratios of zinc to iron would interfere with erythrocyte incorporation of iron in premature infants both during and between feeds. METHODS: In the first experiment, five premature infants (<2500 g birth weight) were enrolled, once receiving full oral feeds by nasogastric tube. They received either high (1200 microg/kg, ratio 4:1) or low (300 microg/kg, ratio 1:1) doses of oral zinc sulfate, together with 300 microg/kg oral 58Fe as chloride in saline with 10 mg/kg vitamin C, between designated feeding periods. Each infant served as its own control and randomly received either high or low doses of zinc or iron and then the alternate dose after 2 weeks. In the second experiment, nine additional premature infants were assigned to the same zinc:iron intake protocol except zinc and iron were given with usual oral feeds (premature formula or human milk) equilibrated before feeding. Iron absorption was measured by the erythrocyte incorporation of 58Fe. RESULTS: High doses of zinc given between feeds significantly inhibited erythrocyte incorporation of iron. 58Fe incorporation (%) with the 1:1 ratio of zinc:iron intake was 7.5 (5.7, 10; geometric mean, -I SD, +1 SD). The percentage of 58Fe incorporation on the 4:1 ratio of zinc:iron intake was 3.6 (2.6, 5.1). Given with feeds, the percentage of 58Fe incorporation on low zinc:iron intake was 7.0 (2.6, 19). Finally, the percentage of 58Fe incorporation on high zinc:iron intake was 6.7 (2.5, 19). CONCLUSION: Elevated intakes of zinc do not interfere with erythrocyte incorporation of iron in premature formulas.

Absorption↗

Ribonuclease activity and isoenzymes in raw and processed cows' milk and infant formulas.

Because of evidence of an immunologic role for ribonuclease II (E.C. 3.1.27.5) in mammals, its presence in milk was further characterized to provide a basis for study of possible contributions of its activity to the health of infants. Isoenzymes of ribonuclease II were quantitatively resolved from milk samples as small as 1 ml or less by chromatography on phosphocellulose. Three isoenzymes detected in bovine milk were the previously reported ribonucleases A and B and a form termed ribonuclease II-1. These isoenzymes were in the ratio of 70:30:1. Form II-1 was unique in its inability to hydrolyze polycytidylate. Bovine colostrum contained 10 to 15 times more ribonuclease II-1 than does milk and three times more total ribonuclease II per unit volume. Human milk contains about 1% the concentration of ribonuclease II found in cows' milk. Ribonuclease II activity in milk was quite stable in the acidic conditions of whey production and during low heat treatments. However, most of its enzymatic activity was lost with high heat treatments. No commercially manufactured milk-based or soybean-based infant formula assayed contained nearly as much ribonuclease activity as either human or bovine milk.

Animals↗