Binding added iron to various milk proteins

Binding of an added radionuclide to milk protein and casein components of cow's milk fractionated by the combination of Sephadex G-150 gel filtration and diethylaminoethyl-cellulose chromatography was determined. Iron-59 labeled ferric chloride was added directly to raw whole milk at a concentration of .02 and 10 ppm isotope and carrier, respectively, and held overnight at 4 C. Five milliliters of the skin milk were chromatographed on a Sephadex G-150 column and fractionated into casein, whey protein, and nonprotein materials. The casein fraction was chromatographed on a diethylaminoethyl-cellulose column and separated into its components, alphas-, beta-, and kappa-caseins. Casein bound about 85% of the added iron in skim milk; of this amount, 72, 21, and 4% were associated with the alphas-, beta, and kappa-caseins.     

A longitudinal study of the protein, nitrogen, and lactose contents of human milk from Swedish well-nourished mothers

The contents of total nitrogen, nonprotein nitrogen, lactose, and individual milk proteins have been determined in human milk from well-nourished Swedish mothers. Breast milk samples from 50 mothers at different stages of lactation (up to 170 days) were collected. Furthermore, three mothers gave samples repeatedly throughout the whole lactation period. The protein content in mature milk was found to be 0.8 to 0.9% by amino acid analysis. The nitrogen content and the contents of the major human milk whey proteins, alpha-lactalbumin and lactoferrin, are very high for the first few days, then decrease rapidly and reach, thereafter, the more slowly declining level of mature milk. Nonprotein nitrogen and the nonspecific milk protein serum albumin are present in constant concentrations throughout lactation. The daily milk volumes were determined and found to be 500 to 600 ml in the very early part and 700 to 800 ml in the later part of the lactation period.     

Effect of psychrotrophic microorganisms on the plasmin system in milk

The psychrotrophic bacteria that are present in raw milk produce heat-stable proteases that affect the plasmin system and, in turn, the quality of the processed milk and other dairy products. Three bacterial strains, Pseudomonas spp. SRM21A and SRM28A and Pseudomonas fluorescens M3/6, were inoculated at a level of approximately 10(3) cfu/ml into reconstituted nonfat dry milk and incubated at 4 degrees C for up to 9 d. From each sample, casein and whey fractions were obtained and electrophoresed to visualize protein hydrolysis and plasmin activity. Colorimetric assays were used to quantify plasmin-related activities. The casein SDS-PAGE gels showed that, by 5 d of incubation, caseinolytic activity was visible in whey fractions of all three strains at the same molecular mass range as commercial bovine plasmin, which was used as a control. Colorimetric assays indicated that plasmin activity in the casein samples decreased as incubation time increased; plasmin activity in the whey samples increased, peaking at 5 to 7 d. Plasminogen activity decreased over time in the casein fraction and increased in the whey fraction. Protein immunodetection indicated crossreactivity between anti-bovine plasminogen and the plasmin and plasminogen from whey samples collected at 5 and 7 d from the incubated milk. The growth of the Pseudomonas strains tested and their concomitant production of proteases caused the release of plasmin and plasminogen from the casein micelle into the whey fraction.     

Indirect and direct determination of the casein content of milk by Kjeldahl nitrogen analysis: collaborative study

The classic method for determination of milk casein is based on precipitation of casein at pH 4.6. Precipitated milk casein is removed by filtration and the nitrogen content of either the precipitate (direct casein method) or filtrate (noncasein nitrogen; NCN) is determined by Kjeldahl analysis. For the indirect casein method, milk total nitrogen (TN; Method 991.20) is also determined and casein is calculated as TN minus NCN. Ten laboratories tested 9 pairs of blind duplicate raw milk materials with a casein range of 2.42-3.05% by both the direct and indirect casein methods. Statistical performance expressed in protein equivalents (nitrogen x 6.38) with invalid and outlier data removed was as follows: NCN method (wt%), mean = 0.762, sr = 0.010, sR = 0.016, repeatability relative standard deviation (RSDr) = 1.287%, reproducibility relative standard deviation (RSDr) = 2.146%; indirect casein method (wt%), mean = 2.585, repeatability = 0.015, reproducibility = 0.022, RSDr = 0.560%, RSDR = 0.841; direct casein method (wt%), mean = 2.575, sr = 0.015, sR = 0.025, RSDr = 0.597%, RSDR = 0.988%. Method performance was acceptable and comparable to similar Kjeldahl methods for determining nitrogen content of milk (Methods 991.20, 991.21, 991.22, 991.23). The direct casein, indirect casein, and noncasein nitrogen methods have been adopted by AOAC INTERNATIONAL.     

Latex allergy: a life-threatening complication

Bone mineralization of healthy preterm infants fed human milk were compared with that of similar fed preterm formula. Bone mineralization was studied by dual energy X-ray absorptiometry in 43 preterm infants divided into two groups; 21 preterm infants were fed with maternal breast milk and 22 preterm infants with a preterm formula containing 70 mg calcium and 35 mg phosphorus per decilitre. CONCLUSION: Preterm infants fed breast milk had lower bone mineral density than the preterm formula-fed group. Fortifying preterm human milk with calcium and phosphorus will improve bone mineralization in preterm infants.     

Standards of pediatric electrophysiology

Milk proteins are hydrolyzed to prevent immunological reactions, but immunoreactive epitopes, including the ABBOS epitope of bovine serum albumin (BSA), can still be detected in commercially available milk protein hydrolysates. We used lactococcal cell-envelope proteinase (CEP) for the hydrolysis of the individual milk proteins and of mixtures thereof, or for the hydrolysis of sodium caseinate (contaminated with whey proteins). CEP exclusively degraded casein, leaving the four major whey proteins intact. This property facilitated the removal of the intact whey proteins from the casein fragments by ultrafiltration. Depending on the molecular mass of the whey protein to be removed, membranes with cutoff values between 3 and 30 kDa were used, resulting in casein hydrolysates free of protein fragments with cross-reactive whey-protein-specific IgE (immunoglobulin E) or ABBOS antibody-binding sites. Even the casein itself was degraded in such a way by CEP that cross-reactive casein-specific IgE antibody-binding sites could be eliminated. The product could find application in infant formulas for therapeutic and preventive treatment of children with cow's milk allergy; in addition, the preventive use of such formulas in children genetically susceptible to the development of insulin-dependent diabetes mellitus (IDDM) should be considered if a relationship between the consumption of BSA and IDDM were to become more apparent. The method is also applicable for preparing casein-free whey protein preparations.     

Breast milk composition in Ethiopian and Swedish mothers. II. Lactose, nitrogen, and protein contents

Breast milk from underprivileged and privileged Ethiopian mothers was collected at different stages of lactation and analyzed for total nitrogen, nonprotein nitrogen, lactose, and individual milk proteins (lactoferrin, alpha-lactalbumin, serum albumin, IgG and IgM). These values and the milk volume of one meal were compared to corresponding results from well-nourished Swedish mothers. No significant differences between the levels of these constituents or the milk volumes were found between the two groups of Ethiopian mothers. When comparison was made between breast milk from these two Ethiopian groups and the Swedish group, the former two showed significantly higher values for the iron-binding protein lactoferrin.     

Exclusive use of calcium channel blockers and cardioselective beta-blockers in the pre- and per-operative management of pheochromocytomas. 70 cases

OBJECTIVE: The influence of liver disease on the pharmacokinetics of candesartan, a long-acting selective AT1 subtype angiotensin II receptor antagonist was studied. METHODS: Twelve healthy subjects and 12 patients with mild to moderate liver impairment received a single oral dose of 12 mg of candesartan cilexetil on day 1 and once-daily doses of 12 mg on days 3-7. The drug was taken before breakfast. Serial blood samples were collected for 48 h after the first and last administration on days 1 and 7. Serum was analyzed for unchanged candesartan by HPLC with UV detection. RESULTS: The pharmacokinetic parameters on days 1 and 7 revealed no statistically significant influence of liver impairment on the pharmacokinetics of candesartan. Following single dose administration on day 1, the mean Cmax was 95.2 ng.ml-1 in healthy subjects and 109 ng.ml-1 in the patients. The AUC0-infinity was 909 ng.h.ml-1 in healthy volunteers and 1107 ng.h.ml-1 in patients and the elimination half-life was 9.3 h in healthy volunteers and 12 h in the patients. At steady state on day 7, mean Cmax values were similar in both groups (112 vs 116 ng.ml-1); the AUC tau was 880 ng.h.ml-1 in healthy subjects and 1080 ng.h.ml-1 in patients while the elimination half-life was 10 h in healthy subjects and 12 h in the patients with liver impairment. The ACU0-infinity on day 1 was almost identical to the AUC tau on day 7. A moderate drug accumulation of 20%, which does not require a dose adjustment, was observed following once-daily dosing in both groups. No serious or severe adverse events were reported. CONCLUSION: Mild to moderate liver impairment has no clinically relevant effect on candesartan pharmacokinetics, and no dose adjustment is required for such patients.     

The effect of sodium prasterone sulfate on lactation

To study the relationship between sodium prasterone sulfate and lactation, blood samples were collected from 120 cases. Sixty cases were in study group (at different periods: during the 3rd trimester, and 1st, 3rd day postpartum) including 30 cases using 100mg sodium prasterone sulfate, 30 cases (using 200mg sodium prasterone sulfate), 60 cases were in control group. The serum levels of estradiol (E2), prolactin (PRL) and plasma levels of oxytocin (OT) were measured by radioimmunoassay, and the production of breast milk was observed. The results showed no statistically significant difference of serum E2, PRL and plasma OT between (sodium prasterone sulfate) study groups and control groups, and the production of breast milk indicated also no significant difference between the 2 groups. The conclusion is that low-dose sodium prasterone sulfate (200mg/day, 600mg/week) did not affect the production of breast milk.     

Nutritional value and antigenicity of two milk protein hydrolysates in rats and guinea pigs

Two milk protein (whey protein and casein) hydrolysates were obtained by enzymatic hydrolysis. In the casein hydrolysate, an ultrafiltration stage was performed to remove all the peptides with molecular weights > 2500. A nutritional value study of both hydrolysates was undertaken. No differences were found between the native proteins and their enzymatic hydrolysates. Neither the enzymatic hydrolysis nor the heat treatments used affected the nutritional value of the protein sources. The two hydrolysates were studied for antigenic properties. The enzymatic hydrolysis of the whey protein concentrate followed by a thermal treatment at 90 degrees C for 10 min reduced significantly its antigenicity. However, the in vivo allergenicity tests showed some positive reactions in both systemic anaphylaxis and passive cutaneous anaphylaxis. The casein hydrolysate, after being ultrafiltered, was devoid of sensitizing capacity by the oral route and was also ineffective in producing local or systemic anaphylaxis in previously sensitized animals, as shown by in vivo assays.     

Effects on nutrient and hormonal profile of long-term infusions of glucose or insulin plus glucose in cows treated with recombinant bovine somatotropin before peak milk yield

Ten Holstein cows were treated with 30.9 mg.d-1 of recombinant bST from 15 to 41 d of lactation. The Latin square design included three infusion periods of 6 d each with 3 d of rest between infusion periods. Infusions were physiological saline, glucose (50 g.h-1), and insulin plus glucose (12.5 IU.h-1 + 50 g.h-1). Blood was collected continuously during the last 24 h of each infusion period. Statistical analyses of data for energy balance, milk yield, and DMI were performed on the last 3 d of each infusion period. Production data before and after infusions (i.e., no recombinant bST) estimated that recombinant bST increased milk yield of cows infused with glucose and saline by 3.1 and 3.6 kg.d-1, respectively. Net energy intake was not affected by infusion, but glucose infusion resulted in higher BW loss than did saline infusion (2.33 vs. 0.08 kg.d-1, respectively), and insulin plus glucose infusion resulted in BW gain (0.65 kg.d-1). Milk yield was 39.9, 39.6, and 37.6 kg.d-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The insulin plus glucose infusion increased milk protein 11 and 14% compared with response to saline and glucose infusions, respectively; no change occurred in the proportion of casein and whey proteins. Serum bST was increased 109% with exogenous recombinant bST. Serum IGF-I was lower for cows infused with glucose than for those infused with saline (21.03 vs. 27.44 ng.ml-1) and increased to 46.55 ng.ml-1 for cows infused with insulin plus glucose. Serum concentrations of insulin and glucose were 13.7 and 56.7, 18.5 and 61.9, and 30.5 muIU.ml-1 and 39.4 mg.dl-1 for cows infused with saline, glucose, and insulin plus glucose, respectively. The results of this study suggest that low concentrations of plasma insulin in early lactation may limit the IGF-I response to recombinant bST (uncoupling). Despite higher IGF-I, milk yield was lower, probably as a result of low blood glucose. These results suggest that, in early lactation, insulin is still anabolic because the BW gain of cows increased. However, milk yield was still higher than that for cows in late lactation with similar insulin concentrations.     

Chromosomal analysis of multipronuclear zygotes obtained after partial zona dissection of the oocytes

OBJECTIVE: To evaluate the hypothesis that breast-feeding women who participate in relaxation training will have increased secretory IgA (sIgA) levels in their breast milk compared with women not receiving training. DESIGN: Nonrandomized control trial of a convenience sample. SETTING: Women were recruited from the postpartum floor of a university teaching hospital. The intervention took place in the women's homes. PARTICIPANTS: Women in the first 48 hours after delivery who were planning to breast-feed their healthy newborn infants for at least 8 weeks were approached for enrollment. Women were excluded if they had previous experience with relaxation training. At 4 to 6 weeks postpartum, we enrolled 38 women still breast-feeding their infants. INTERVENTIONS: Women were allocated into 3 groups. Women in group 1 were taught relaxation and had breast milk samples collected before and after the teaching. Women in group 2 had conversation with similar breast milk sample collection, and women in group 3 had 1 breast milk sample collected. Women in group 1 were encouraged to practice the relaxation once or twice a day for 2 weeks, and a second visit was made to all mothers with repeated breast milk collections. Women who were still breast-feeding at 6 to 8 weeks after study end had a final breast milk sample collected. Breast milk was analyzed for secretory IgA levels. Stress was assayed using the Symptom Checklist-90-R and open-ended questions. RESULTS: There was no difference in sIgA levels among the 3 groups at any time. Women who reported stress present between visit 1 and visit 2 increased their sIgA levels at the final sample collection (+0.16 g/L) compared with women who reported no stress (-0.09 g/L; P= .03). The ratings of success in relaxation in women in group 1 were related to the following sIgA levels in sample 4: poor relaxation, 0.67 g/L; fair relaxation, 0.41 g/L; good relaxation, 0.35 g/L; and very good, 0.30 g/L (P= .006). CONCLUSIONS: Self-reported stress appears to increase breast milk sIgA levels. Success at relaxation was inversely related to sIgA levels in the group learning relaxation.     

The relative paucity of IgE in human milk

The levels of IgE were determined in paired samples of serum and milk when whey obtained 3 to 8 days of postpartum, from 16 human lactating mothers who had reported a history of allergy to a variety of common allergens. Two assay procedures were employed to measure total IgE, a double-antibody assay and a commercially available solid phase assay (RIST). In addition, each sample of serum and whey was tested for specific IgE antibodies to a variety of allergens by the RAST test. The levels of total serum IgE were between 30 and 2300 I.U./ml and relatively good agreement was observed for both the double-antibody and RIST methods. In contrast, total IgE levels in milk whey were either undetectable (less than 3.0 I.U./ml in 14 of 16 subjects) or very low when analyzed by the double-antibody method, but were very high (400 to 1650 I.U./ml when analyzed by the RIST method. However, IgE added to milk whey could be measured by the double-antibody procedure indicating that the low levels detected in milk were not a fault of the double-antibody assay. It was assumed that the RIST test was subject to nonspecific interference by factors in milk whey which caused the determination of high, but incorrect, levels of IgE. Specific IgE antibodies were detected in the serum of 10 of 16 subjects but were not present in milk whey. A comparison of the whey/serum ratios of albumin, IgA, and IgE suggested that little, if any, IgE is selectively synthesized or secreted in the mammary gland.     

Use of urea by early postpartum Holstein cows

Thirty-nine lactating Holstein cows were fed high-energy complete rations ad libitum with crude protein: 1) 11.7% (negative control); 2) 13.9% (1% urea); 3) 16.6% (1% urea); or 4) 16.6% (positive control) in a continuous 12-wk study beginning at wk 5 postpartum. Milk production of 27.7, 31.8, 34.0, and 30.4 kg/day showed the use of urea nitrogen by groups 2 and probably 3. Two digestion-nitrogen balance trials with each cow also provided evidence that urea nitrogen was used for milk secretion. Energy digestibility averaged 59.4, 64.2, 65.4, and 65.8; and lower for the negative control diet. Nitrogen solubility in the diets was 28, 36, 32, and 21%, which reflects the objective of selecting ingredients with low nitrogen solubility for use in urea diets. Concentrations of ammonia nitrogen before and after feeding were 1.1, 3.3, 3.5, 4.2, and 2.2, 11.2, 11.9, and 9.3 mg/100 ml of rumen fluid. The prefeeding amounts were probably too low for maximum microbial growth. Urea-nitrogen concentrations in plasma were 8.65, 10.32, 18.00, and 17.03 mg/100 ml. These results lend support to the postulate that lactating cows in early lactation can use urea nitrogen when high-energy complete rations with ingredients of low nitrogen solubility are fed ad libitum.     

Age- and gender-related elastin distribution changes in human vocal folds

To elucidate the mechanism of gut hypertrophy observed in rats artificially reared (AR) on milk formulas, the effects of four refined formulas with different ratios of casein (C) and whey protein (W), CW 2:8, CW 4:6, CW 6:4 and CW 8:2, on the gut growth of AR rats were examined. Four groups of pups were infused with each formula through an intragastric cannula from age 5 to 15 days. Each of the four milk formulas showed a different character in the stomach, such as no curd, very soft curd, soft curd and hard curd, in response to an increasing ratio of C:W. There were no significant differences in body weight gain among the AR groups and mother-reared (MR) controls. The stomach growth, in weight, of AR rats increased in response to the increasing ratios of C:W. In comparison with MR controls, hypertrophy of the stomach of AR rats appeared within the formulas with higher proportions of casein than whey protein (CW 6:4 and CW 8:2), but not those with lower proportions (CW 2:8 and CW 4:6). The growth of the small intestine was also related to the increasing ratio of C:W in the formulas. A similar pattern of hypertrophy in the hindgut was seen in AR rats. There was no association between hypertrophy of the gut in AR rats and plasma triiodothyronine. The present results clearly demonstrated that the gut growth of AR rat pups was directly influenced by the diet but not by AR per se, and that hard casein-curd in the stomach might be one cause of gut hypertrophy.     

Ingredient supplementation effects on viability of probiotic bacteria in yogurt

The present investigation studied the effects of cysteine, whey powder, whey protein concentrate, acid casein hydrolysates, or tryptone on the viability of Streptococcus thermophilus, Lactobacillus acidophilus, and bifidobacteria. Changes in pH, titratable acidity, redox potential, and viability of bacteria were monitored during 24 h of fermentation and refrigerated storage (4 degrees C) of yogurt for 35 d. The incubation time that was needed to reach pH 4.5 was considerably affected by the added ingredients. Also, the drop in pH or the increase in acidity and redox potential was dependent on the added ingredients. The addition of cysteine, whey protein concentrate, acid casein hydrolysates, or tryptone improved the viability of bifidobacteria to a variable extent, but whey powder failed to improve their viability. The morphology of S. thermophilus, as shown by electron microscopy, was affected by cysteine at 500 mg/L, possibly as a result of reduced redox potential. Sodium dodecyl sulfate-PAGE and amino acid analyses suggested that the nitrogen source in the form of peptides and amino acids improved the viability of bifidobacteria in yogurt made with a commercial ABT (Lactobacillus acidophilus, bifidobacteria, and Streptococcus thermophilus) starter culture, which showed a dramatic decline in the counts of this organism in previous studies.     

Biochemical markers of calcium and bone metabolism during 18 months of lactation in Gambian women accustomed to a low calcium intake and in those consuming a calcium supplement

The effect of 18 months of lactation on indexes of calcium and bone metabolism was studied in 60 Gambian women accustomed to a very low calcium intake. Half the women consumed a calcium supplement from 10 days postpartum for 52 weeks (supplement, 714 mg Ca/day; total Ca intake, 992 +/- 114 mg/day), and half consumed placebo (total Ca intake, 288 +/- 128 mg/day). Fasting blood and 24-h urine samples were collected at 1.5, 13, 52, and 78 weeks of lactation and analyzed for calciotropic hormones (intact PTH, 1,25-dihydroxyvitamin D, and calcitonin), bone turnover markers (osteocalcin, bone alkaline phosphatase, and urinary deoxypyridinoline), and plasma minerals (calcium and phosphate). The first months of lactation were associated with increased bone turnover and plasma phosphate, and decreased PTH and 1,25-dihydroxyvitamin D. These effects diminished by 52 weeks, although breast milk volumes remained high. The Gambians had higher PTH, 1,25-dihydroxyvitamin D, and bone formation than British women with a greater customary calcium intake. None of the biochemical indexes was affected by calcium supplementation, with the possible exception of bone alkaline phosphatase (-29% at 52 weeks; P = 0.015). These data demonstrate that lactation-associated changes in calcium and bone metabolism are physiological and are independent of dietary calcium supply in women with very low calcium intakes.     

Ultrasound velocity of trabecular cubes reflects mainly bone density and elasticity

We examined hepatic cytochrome P450 (CYP) induction in rat foetuses and neonates by phenobarbital administered through placenta or breast feeding. In an intraperitoneal study, phenobarbital was administered intraperitoneally to mother rats once a day for 7 consecutive days before delivery. The livers were removed from foetuses, neonates, and mothers just before and 5 and 10 days after delivery. In oral administration study, water containing phenobarbital was given orally ad libitum from day 13 of pregnancy to 3 weeks after delivery (end of lactation). The livers were removed from neonates and mothers just before and one week after delactation. Phenobarbital administered intraperitoneally increased both the activity and the protein expression of CYP2B in 5-day-old neonates, even though the administration ended before delivery. This increase had disappeared in 10-day-old neonates. In mother rats, phenobarbital increased CYP2B just before and 5 days after delivery, while no increase was detected 10 days after delivery. Phenobarbital administered orally also increased both the activity and the protein expression of CYP2B of neonates and mothers during lactation and this increase also disappeared 1 week after delactation. Neither activity nor protein expression of CYP3A were induced in perinates at any age examined in either administration route. In mother rats, increase in CYP3A was found only just before delivery in the peritoneal administration study. Our results suggest that phenobarbital administered through placenta or breast milk transiently induces hepatic CYP2B in newborn rats but that the influence of phenobarbital does not last long.     

Evidence for defective skeletal mineralization in low-birthweight infants: the absorption of calcium and fat

Using serial metabolic balance techniques, the absorption and retention of calcium and the absorption of fat have beem measured over the first 30 to 70 days of life in 11 preterm and 2 full-term light-for-dates infants. They were fed either full-cream cow's milk, half-skimmed cow's milk, the proprietary filled milk S.M.A., or breast milk. The values for calcium intake, absorption and retention were compared with the rate of accumulation of calcium by the fetus in utero, which was calculated from published data on the chemical composition of fetal bodies. Infants fed breast milk had an absolute dietary deficiency of calcium. Those fed other milks ingested sufficient but they did not absorb enough. Though calcium absorption increased with increasing postnatal age, intrauterine rates of calcium retention were never achieved on any of the milks. The average retention of calcium by preterm infants as a percentage of intrauterine accumulation was, for cow's milk 38%, for S.M.A. 27%, and for breast milk 17%. The full-term light-for- dates infants absorbed and retained more calcium than the preterm infants; it was on average 52% of the amount accumulated by the human fetus for an equivalent weight gain. The average absorption of fat by preterm infants was, from the cow's milk preparations 55%, from S.M.A. 61%, and from breast milk 84%. The light-for-dates infants absorbed on average 87% of the breast milk fat. There was no evidence that the amount of calcium absorbed was materially influenced by fat malabsorption. The principal determinants of the amount of calcium absorbed were the length of gestation and postnatal age of the infant.