n-acetyl-beta-hexosaminidase activity in human breast milk

1. The lysosomal enzyme, N-acetyl-beta-hexosaminidase (HEX) is present in human breast milk. It is composed predominantly of "A" (heat-labile) and "B" (heat-stable) isozymes which coelute with the corresponding major serum isozymes on DE-52 ion-exchange chromatography. 2. Total HEX activity in "early" milk obtained at 2.8 +/- 1.4 weeks post partum, is approx. 2.5-fold higher (87 +/- 29 nmol/60'/mg protein. n = 10) than that of pregnancy serum (35.7 nmol/60'/mg protein) prior to delivery. 3. These levels increase to greater than 3-fold (110 +/- 20 nmol/60'/mg protein, n = 13) as the milk matures (10.3 +/- 4.2 weeks). 4. The specific activity of HEX A in the milk changes little with time post partum, because absolute 5. In contrast, HEX B specific activity is increased, as absolute levels (per volume) remain constant in the face of decreasing milk protein content, 5. These changes result in a high degree of correlation (r = 0.81) between time of lactation and % HEX A observed.     

Docosahexaenoic acid transfer into human milk after dietary supplementation: a randomized clinical trial

Docosahexaenoic acid (DHA) is important for infant development. The DHA transfer from maternal diet into human milk has not been investigated in detail. We studied the effects of DHA supplementation on the fatty acid composition of human milk and the secretion of dietary (13)C-labeled fatty acids, including DHA, into human milk. Ten lactating women were randomized to consume, from 4 to 6 weeks postpartum, an oil rich in DHA (DHASCO, 200 mg of DHA/day) (n = 5) or a placebo oil (n = 5). Dietary intakes were followed by 7-day protocols. On study day 14 a single dose of [U-(13)C]DHASCO was given orally, milk samples were collected over 48 h, and milk production was recorded. Milk fatty acid composition was determined by gas-liquid chromatography and isotopic enrichment was determined by gas chromatography- combustion-isotope ratio mass spectrometry (GC-C-IRMS). Milk DHA content did not differ between the supplemented and placebo group at study entry (0.29 vs. 0.28 wt%, median). After 2 weeks of supplementation the milk DHA content was almost 2-fold higher in the supplemented versus placebo group (0.37 vs. 0.21 wt%, P = 0.003). Cumulative recovery of [(13)C]palmitic, [(13)C]oleic, and [(13)C]docosahexaenoic acids in human milk at 48 h was similar between supplemented and placebo groups (palmitic acid 7.40 vs. 8. 14%, oleic acid 9.14 vs. 9.97%, and docosahexaenoic acid 9.09 vs. 8. 03% of dose, respectively). Notable lower recovery was observed for [(13)C]myristic acid in both the supplemented and placebo groups, 0. 62 versus 0.77% of dose.Dietary DHA supplementation increases the DHA content in human milk. DHA transfer from the diet into human milk is comparable to palmitic and oleic acid transfer.     

The breast cancer resistance protein BCRP (ABCG2) concentrates drugs and carcinogenic xenotoxins into milk

Contamination of milk with drugs, pesticides and other xenotoxins can pose a major health risk to breast-fed infants and dairy consumers. Here we show that the multidrug transporter BCRP (encoded by ABCG2) is strongly induced in the mammary gland of mice, cows and humans during lactation and that it is responsible for the active secretion of clinically and toxicologically important substrates such as the dietary carcinogen PhIP, the anticancer drug topotecan and the antiulcerative cimetidine into mouse milk.     

Isolation and characterization of human breast milk lipoamidase

The mean lipoamidase activity in human breast milk was found to be 0.073 nmol/min per mg (S.D. = 0.028, range = 0.020-0.123, n = 44). The mean lipoamidase activity is approximately 3-fold higher in milk than that in serum (0.023 nmol/min per mg, S.D. = 0.016, range = 0.001-0.059, n = 32). Lipoamidase was purified to 4400-fold by a four-step procedure from 330 ml of human breast milk. The purified enzyme was identified as a single band (Mr = 135,000) by sodium dodecyl sulfate/polyacrylamide electrophoresis. Analysis by Edman degradation indicated that the N-terminal amino acid was glycine. These results strongly suggest that milk lipoamidase is composed of a single polypeptide chain. The enzyme is considered to be a glycoprotein since it reacted positively to periodate-Schiff (PAS) staining. The isoelectric point of the enzyme was 4.2. After treatment of lipoamidase with sialidase, its position on isoelectric focusing gel moved from pH 4.2 to 4.6. This is strongly indicative that lipoamidase contains sialic acid residues. The optimum pH for the enzyme activity is 7.0. The Michaelis constant (KM) for lipoyl p-aminobenzoate is calculated as 25 microM. The enzyme activity was completely lost by heating 60 degrees C for 5 min. The effects of thiol-reactive agents, such as 2-mercaptoethanol (ME) and p-chloromercuribenzoate, were not significant. However, the enzyme activity was completely inhibited by 50 microM diisopropylfluorophosphate. Thus, this enzyme seemed to contain an essential serine residue in the active site.     

The transfer of tritium-labeled organic material from grass into cow's milk

Two lactating cows were given tritiated hay containing organically bound tritium (OBT) only for about 4 weeks. Tritium activity was determined in milk fat, casein, lactose, milk water, and whole milk. In one cow, milk was sampled for approximately 450 days, covering two lactation periods. At steady state, specific tritium activities in casein, lactose, and milk water were 58, 10, and 11%, respectively, of those in milk fat. Some OBT was converted into THO during catabolism and entered the body water pool. This 3H source accounted for nearly 40% of tritium in lactose, but in casein and milk fat about 97% of tritium was derived from ingested OBT. Comparison of the specific activity of milk constituents with the specific activity of ingested hay showed the following values: 0.84 for milk fat, 0.49 for casein, 0.05 for lactose, 0.10 for milk water. About one-half of the tritium transferred to milk was found in organic milk constituents and the other half in milk water. Decrease of tritium activity with time could be represented by three components with different half-lives for the organic milk constituents. Those for milk fat and casein were quite similar, with a slow component of nearly 3 months.     

A comparative study of elemental composition of human breast milk and infant milk substitutes

Instrumental neutron activation analysis (INAA) and protoninduced X-ray emission (PIXE) analysis have been employed to determine the concentration of 13 elements in human breast milk, various infant formulas, and locally produced cereals from Nigeria, as well as from various infant formulas and natural cow and goat milk available in the UK. The study shows that if the locally produced cereal is to be used on a regular basis for babies in Nigeria, then their diet must be supplemented with essential trace elements. Furthermore, parents should be discouraged from giving their infants cow and goat milk because of the high concentration of major elements compared to human breast milk.     

Determination of hyaluronan molecular mass distribution in human breast milk

Hyaluronan (HA) in human milk mediates host responses to microbial infection via TLR4- and CD44-dependent signaling. Signaling by HA is generally size specific. Because pure HA with average molecular mass (M) of 35 kDa can elicit a protective response in intestinal epithelial cells, it has been proposed that human milk HA may have a bioactive low-M component. Here we report the size distribution of HA in human milk samples from 20 unique donors. A new method for HA analysis, employing ion exchange (IEX) chromatography to fractionate HA by size and specific quantification of each size fraction by competitive enzyme-linked sorbent assay (ELSA), was developed. When separated into four fractions, milk HA with M ? 20 kDa, M ∼ 20 to 60 kDa, and M ∼ 60 to 110 kDa comprised averages of 1.5, 1.4, and 2.0% of the total HA, respectively. The remaining 95% was HA with M ? 110 kDa. Electrophoretic analysis of the higher M HA from 13 samples showed nearly identical M distributions, with an average M of approximately 440 kDa. This higher M HA component in human milk is proposed to bind to CD44 and to enhance human beta defensin 2 (HBD2) induction by the low-M HA components.     

Measurement of cerium in human breast milk and blood samples

The aim of this study was to evaluate the relationship between cerium content in human breast milk and blood plasma or serum. Blood samples and breast milk at various stages of lactation, from 5 days to 51 weeks post partum, were donated by 42 healthy breast-feeding mothers from Munich, Germany and by 26 lactating Spanish mothers from Madrid at 4 weeks post partum. Inductively coupled plasma mass spectrometry was applied for the determination of cerium in the biological samples. Cerium concentration in the digested milk samples from Munich showed low values and the arithmetic mean values ranged between the quantification limit of 5 ng/L up to 65 ng/L. The median value amounted to 13 ng/L. The cerium concentrations in the Spanish breast milk samples amounted to similar low values. The data were about a factor of eight lower than values found in a former study of samples from an eastern German province. All cerium concentrations in the German plasma samples, except for two, were at the quantification limit of 10 ng/L. Interestingly, the serum samples of the Spanish mothers showed cerium values ranging between 21.6 and 70.3 ng/L; these higher data could be explained by an enhanced intake of cerium by humans in Madrid. This could be caused by increased cerium concentrations in particulate matter due to a higher traffic volume in Madrid compared to Munich.The results obtained in this study contribute to setting reference baseline values of cerium in human breast milk and blood plasma/serum and indicate a varying cerium amount depending on the cerium environmental pollution. Possibly, the cerium content in plasma/serum could be an indicator for environmental cerium, which is not valid for breast milk.     

High risk human papillomavirus and Epstein Barr virus in human breast milk

ABSTRACT: BACKGROUND: Multiple viruses, including human immunodeficiency virus, Epstein Barr virus (EBV) and mouse mammary tumour virus have been identified in human milk. High risk human papillomavirus (HPV) sequences have been identified in breast cancer. The aim of this study is to determine if viral sequences are present in human milk from normal lactating women. FINDINGS: Standard (liquid) and in situ polymerase chain reaction (PCR) techniques were used to identify HPV and EBV in human milk samples from normal lactating Australian women who had no history of breast cancer.High risk human papillomavirus was identified in milk samples of 6 of 40 (15%) from normal lactating women - sequencing on four samples showed three were HPV 16 and one was HPV 18. Epstein Barr virus was identified in fourteen samples (33%). CONCLUSION: The presence of high risk HPV and EBV in human milk suggests the possibility of milk transmission of these viruses. However, given the rarity of viral associated malignancies in young people, it is possible but unlikely, that such transmission is associated with breast or other cancers.     

Epidermal growth factor-like proteins in breast fluid and human milk

Epidermal growth factor (EGF), and the transforming growth factor-alpha (TGF-alpha) family of proteins, which also bind to the EGF receptor, have been associated with human breast cancer. The total EGF-like proteins were determined by a radioreceptor assay, and TGF-alpha by radioimmunoassay, in human milk and breast fluid samples. The breast fluids were collected by nipple aspiration from healthy premenopausal women. Both the 24 milks and 18 breast fluids assayed contained EGF-like proteins, at concentrations ranging from 32-600 ng/ml (median, 140 ng/ml), and 62-654 ng/ml (median, 205 ng/ml) respectively. Immunoreactive TGF-alpha proteins were detected at higher levels in 21 breast fluids (range, 0-50.0; median 5.1 ng/ml) than in 24 milk samples (range, 0-8.4; median, 0.8 ng/ml).     

Dynamics of human milk extraction: A comparative study of breast feeding and breast pumping

We describe a mathematical model of the flow and deformation in a human teat. Our aim is to compare the theoretical milk yield during infant breast feeding with that obtained through the use of a breast pump. Infants use a peristaltic motion of the tongue, along with some suction, to extract milk, whereas breast pumps use a cyclic pattern of suction only. Our model is based on quasi-linear poroelasticity whereby the teat is modelled as a cylindrical porous elastic material saturated with fluid. We impose a cyclic axial suction pressure difference across the teat and impose a radial compressive force moving along the teat which mimics infant suckling. This is compared to the case of cyclic and steady pumping only which models the action of breast pumps. The results illustrate that there is an optimal time to apply the compressive force during the suction cycle that will increase the flow rate in our theoretical teat. The model and results may be of use in the future design of effective breast pumps.     

Influence of stable iodine on the transfer of131I into cows' milk

The effect of supplying three different stable iodine levels (about 70, 80, and 160 mg day^–1) in the daily feed of four dairy cows has been studied for the excretion pattern of¹³¹I. None of the different treatments had a statistically significant effect. A mean milk transfer coefficient of 0.015 ± 0.001 d 1^–1 was determined for all iodine supply levels investigated.