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PubMed1981

Chemical and physical characterization of a proline-rich polypeptide from sheep colostrum.

A proline-rich polypeptide isolated from sheep colostrum is described. The molecular weight of the polypeptide determined by gel filtration is 17 200. However, in the presence of guanidinium chloride the molecular weight found is about 6000. The polypeptide contains about 22% of proline, a high proportion of non-polar amino acids, a low percentage of glycine, and no alanine, arginine and cysteine residues. The only N-terminal amino acid found is leucine. C.d. spectra in water and in 50% (v/v) trifluoroethanol suggest the presence of block sequences of proline residues forming helices of polyproline II type. The proline-rich polypeptide is soluble at 4 degrees C but is reversibly precipitated on warming to room temperature. Maximal precipitation is observed at pH 4.6 and at ionic strength above 0.6. The precipitation depends on the concentration of the polypeptide. No effect of other proteins, Ca2+ and Zn2+ ions on the precipitation of the polypeptide was found. The proline-rich polypeptide is not an amphipathic protein. The lack of effect of the polypeptide on proteolytic enzymes ruled out the possibility that it is an inhibitor of proteinases.

Amino Acids↗

PubMed1988

Insulin-like growth factors 1 and 2 in bovine colostrum. Sequences and biological activities compared with those of a potent truncated form.

1. Insulin-like growth factors 1 and 2 (IGF-1 and IGF-2) together with a truncated form of IGF-1 were purified to homogeneity from bovine colostrum. 2. Two forms of IGF-1 were totally resolved from IGF-2 in the purification by h.p.l.c. involving cation-exchange and reverse-phase columns. 3. The complete amino acid sequences for all three forms of IGF were determined. The sequence of bovine IGF-1 was found to be identical with that of human IGF-1, and that of the variant lacked the N-terminal tripeptide Gly-Pro-Glu (-3N:IGF-1). Bovine IGF-2 was found to differ in three residues of the C-domain compared with human IGF-2, with serine, isoleucine and asparagine substituted for alanine, valine and serine respectively at positions 32, 35 and 36. 4. Protein synthesis in L6 rat myoblasts was stimulated and protein degradation inhibited in a co-ordinate response with all three IGFs. The relative potency in both processes was -3N:IGF-1 greater than IGF-1 greater than IGF-2. A similar order of potency was obtained for the stimulation of DNA synthesis by -3N:IGF-1 and IGF-1. The approximately 10-fold effect on biological activity of removing the N-terminal tripeptide is unexpected in view of current information on IGF-1 structure and function.

Amino Acid Sequence↗

PubMed1992

Bovine colostrum CMP-NeuAc:Gal beta(1-->4)GlcNAc-R alpha(2-->6)-sialyltransferase is involved in the synthesis of the terminal NeuAc alpha(2-->6)GalNAc beta(1-->4)GlcNAc sequence occurring on N-linked glycans of bovine milk glycoproteins.

Bovine colostrum CMP-NeuAc:Gal beta(-->4)GlcNAc-R alpha(2-->6)-sialyltransferase (alpha 6-sialyltransferase) appears to be capable of catalysing alpha 6-sialylation of the disaccharide GalNAc beta(1-->4)GlcNAc to yield the trisaccharide NeuAc alpha(2-->6)GalNAc beta(1-->4)GlcNAc. This provides an enzymic basis for the occurrence of this sialylated structure on the N-linked glycans of a number of bovine milk glycoproteins. Competition experiments using Gal beta(1-->4)GlcNAc and GalNAc beta(-->4)GlcNAc as acceptors indicate that both substrates are recognized by a single active site on the alpha 6-sialyltransferase. Extrapolation of these results suggests that the NeuAc alpha(2-->6)GalNAc beta(1-->4)GlcNAc structural element occurring on the N-linked glycans of several human glycoproteins are similarly synthesized by the action of a Gal beta(1-->4)GlcNAc-R alpha(2-->6)-sialyltransferase.

Animals↗

PubMed1999

Effect of proline rich polypeptide from ovine colostrum on virus replication in human placenta and amniotic membrane at term; possible role of endogenous tumour necrosis factor alpha.

Freshly prepared organ cultures of human placentae and amniotic membranes at term show different sensitivity to vesicular stomatitis virus (VSV) infection. In six of 16 amniotic membranes and seven of 17 placentae VSV replicated to relatively high titres (10(3)-10(6)TCID(50)/ml). The others were partially or completely resistant to virus infection (<10(1)-10(2)TCID(50)/ml). Addition of the immunomodulating agent, proline-rich-polypeptide (PRP) from ovine colostrum to explants freshly obtained from the organs, influenced VSV replication in a manner dependent on the innate immune state of the organ culture. In cultures resistant to the virus, PRP at a concentration of 10 microg/ml increased 10-10 000 times the VSV titre. In contrast, treatment of highly sensitive cultures by PRP hardly influenced viral replication at all. The effect of virus stimulation by PRP was abolished by specific anti-TNF antibodies. The results indicate that endogenous TNF may be one of the mediators of virus stimulation by PRP. Antibodies against TNFalpha, added to VSV infected organ cultures sensitive to the virus reduced viral replication. The antibodies caused stimulation of virus replication in VSV-infected resistant organ cultures. The results indicate the double role of endogenous TNF in viral replication in placenta and the amniotic membrane.

Amnion↗

PubMed1984

The yield and nutrient content of colostrum and milk of women from giving birth to 1 month post-partum.

The intake of mammary secretion from delivery to day 5 post-partum was determined by test-weighing nine infants using an integrating electronic balance. The mean yield of colostrum for the first 24 h after birth was 37.1 (range 7.0-122.5) g and was 408 (range 98.3-775) and 705.4 (range 452.5-876) g/24 h on days 3 and 5 post-partum respectively. The milk yield of mothers on either day 14 or 28 post-partum was determined by test-weighing the mother. The mean milk yield was 1.156 (SD 0.167) kg/24 h. A significant correlation (P less than 0.001; r 0.85, n 42) was found between milk yield measured by test-weighing the infant and milk yield measured by test-weighing the mother, confirming that it is possible to obtain a similar estimate of milk consumed using either of the two methods of test-weighing. There was a significant positive correlation (P less than 0.001) between lactose concentration and milk yield for the first 5 d post-partum (r 0.76, n 22); a significant correlation (P less than 0.001) between protein concentration and milk yield (r-0.74, n 22) and no significant correlation between fat concentration and milk yield for the period studied. The calculated energy intake of infants during the first 24 h after birth was only 0.12 (range 0.02-0.29) mJ. This increased to 1.44 (range 0.83-2.18) and 2.99 (range 2.49-4.06) mJ/24 h by days 3 and 14-28 post-partum respectively.

Body Weight↗

PubMed1993

Thrombocytopenia in lambs fed with bovine colostrum.

Artificially reared lambs, fed with bovine colostrum, died within 48 hours after birth, showing thrombocytopenia and extensive haemorrhages on autopsy. The mechanism behind was not fully understood, but experimental immunization of young cattle against sheep red blood cells, carried out five years earlier on the same farm, may have played a role.

Animal Husbandry↗

PubMed2003

A proline-rich polypeptide complex (PRP) from ovine colostrum. Studies on the effect of PRP on nitric oxide (NO) production induced by LPS in THP-1 cells.

A proline-rich polypeptide complex (PRP) isolated from ovine colostrum shows immunoregulatory activity. Similar activity was observed when PRP was replaced with a nonapeptide (NP) isolated from chymotryptic digest of PRP. The polypeptide complex also shows procognitive activity. In the form of orally administered tablets called Colostrinin, containing 100 microg of PRP, it improves the outcome of Alzheimer's disease (AD) patients. The mechanism of action of PRP/Colostrinin in AD is not yet clarified. Microglial cells involvement in AD has been related to amyloid beta (Abeta) internalization, the release of inflammatory cytokines, overproduction of nitrogen oxide (NO) and superoxide anion (O2-), and the development of neuritic plaques. It has been previously found in our laboratory that PRP regulates the secretion of an array of cytokines. It also was shown, in preliminary experiments using human blood cells and murine macrophages, that PRP inhibits production of NO and O2- induced by LPS. In the present work, to study the effect of PRP and NP on the release of NO and O2-induced by LPS we applied THP-1 cells. The human monocyte/macrophage THP-1 cell line has been widely used as a model of human microglial cells. The results obtained showed that THP-1 cells release NO when activated with LPS. However, neither PRP nor NP induced production of NO. Although the nonapeptide, at higher concentration (100 microg/mL), showed an inhibitory activity on the release of NO induced by LPS, no inhibition was observed when PRP was used. THP-1 cells treated with LPS, PRP or NP did not release O2-.

Adjuvants, Immunologic↗

PubMed1977

Enzymic characteristics of fat globule membranes from bovine colostrum and bovine milk.

Fat globule membranes have been isolated from bovine colostrum and bovine milk by the dispersion of the fat in sucrose solutions at 4 degrees C and fractionation by centrifugation through discontinuous sucrose gradients. The morphology and enzymic characteristics of the separated fractions were examined. Fractions comprising a large proportion of the total extracted membrane were thus obtained having high levels of the Golgi marker enzymes UDP-galactose N-acetylglucosamine beta-4-galactosyltransferase and thiamine pyrophosphatase. A membrane-derived form of the galactosyltransferase has been solubilized from fat and purified to homogeneity. This enzyme is larger in molecular weight than previously studied soluble galactosyltransferases, but resembles in size the galactosyltransferase of lactating mammary Golgi membranes. In contrast, when fat globule membranes were prepared by traditional procedures, which involved washing the fat at higher temperatures, before extraction, galactosyltransferase was not present in the membranes, having been released into supernatant fractions, When the enzyme released by this procedure was partially purified and examined by gel filtration, it was found to be of a degraded form resembling in size the soluble galactosyltransferase of milk. The release is therefore attributed to the action of proteolytic enzymes. Our observations contrast with previous biochemical studies which suggested that Golgi membranes do not contribute to the milk fat globule membrane. They are, however, consistent with electron microscope studies of the fat secretion process, which indicate that secretory vesicle membranes, derived from the Golgi apparatus, may provide a large proportion of the fat globule membrane.

Animals↗

PubMed1998

Prophylactic effect of bovine anti-Cryptosporidium hyperimmune colostrum immunoglobulin in healthy volunteers challenged with Cryptosporidium parvum.

Bovine hyperimmune anti-Cryptosporidium colostrum immunoglobulin (BACI) decreases the intensity of Cryptosporidium parvum infection in vitro. We investigated the prophylactic effect of BACI in healthy adults challenged with C. parvum. After we established an oocyst dose that resulted in 100% infection in four volunteers (baseline group), 16 volunteers were randomized to receive (1) BACI prior to C. parvum challenge (BACI group) and a nonfat milk placebo 30 minutes later, (2) BACI prior to and 30 minutes after challenge (reinforced BACI group), or (3) nonfat milk placebo prior to and 30 minutes after challenge. Subjects received BACI (10 g) or nonfat milk placebo three times a day for a total of 5 days and were followed for clinical symptoms and oocyst excretion for 30 days. A trend toward less diarrhea (P = .08) was observed for subjects receiving BACI in comparison with occurrences in placebo recipients. Subjects receiving BACI or nonfat milk placebo had a 100-fold reduction in oocyst excretion as compared with excretion in the baseline group.

Administration, Oral↗

PubMed2005

The effect of IFN-gamma and TNF-alpha on the NADPH oxidase system of human colostrum macrophages, blood monocytes, and THP-1 cells.

The aim of this work was to analyze the effect of Interferon-gamma (IFN-gamma) and tumor necrosis factor-alpha (TNF-alpha) on NADPH oxidase activity and gp91-phox gene expression in human colostrum macrophages (CM), peripheral blood monocytes (PBM), and myelomonocytic THP-1 cells. We also investigated the effect of IFN-gamma on the release of TNF-alpha by these cells. Our results show that under basal culture conditions, CM release more superoxide than PBM and THP-1 cells (p < 0.05). The addition of IFN-gamma, alone or in combination with TNF-alpha, increased spontaneous superoxide release by PBM and THP-1 cells (p < 0.05) and increased phorbol myristate acetate (PMA)-stimulated superoxide release by CM, PBM, and THP-1 cells (p < 0.05). The NADPH oxidase activity of THP-1 cells consistently remained lower than that of CM or PBM, despite a dramatic response to IFN-gamma and TNF-alpha. Under basal conditions, gp91-phox gene expression was significantly higher in CM and PBM compared with THP-1 cells (p < 0.05). The addition of IFN-gamma alone or in combination with TNF-alpha caused a dramatic increase in gp91-phox gene expression in THP-1 cells (p < 0.05) but not in CM or PBM. Under basal conditions or in the presence of IFN-gamma, CM released more TNF-alpha than PBM or THP-1 cells (p < 0.05). In addition, PBM released more TNF-gamma than THP-1 cells (p < 0.05). IFN-gamma did not significantly augment the release of TNF-alpha by these cells (p > 0.05). Thus, IFN-gamma and TNF-alpha induced equivalent gp91-phox gene expression in THP-1 cells compared with CM or PBM but did not bring about equivalent NADPH oxidase activity. TNF-alpha release was higher in more mature cells. This partial divergence of gp91- phox gene expression, NADPH oxidase activity, and TNF-alpha release is probably a consequence of different events of myeloid cell biology and relates at least in part to cell differentiation state.

Cell Line↗

PubMed1981

Fatty acid composition of human colostrum and mature breast milk.

The fatty acid composition of human milk obtained on individual samples from 120 mothers early (day 3 to 5) and later (day 40 to 45) in lactation were determined by argentation thin-layer and gas chromatographic procedures. In comparison with mature milk, human colostrum was characterized by a lower percentage of saturated fatty acids including medium chain length acids, a higher percentage of monounsaturates, and a lower level of linoleic and linolenic acids, but a higher percentage of their long chain polyunsaturated derivatives. It is concluded that in view of their levels in breast milk, the polyenoic derivatives of linoleic and linolenic acids must be taken into account when assessing infant foods.

Chromatography, Gas↗

PubMed1991

Inhibition of localized adhesion of enteropathogenic Escherichia coli to HEp-2 cells by immunoglobulin and oligosaccharide fractions of human colostrum and breast milk.

Secretory IgA (sIgA) purified from colostrum and breast milk obtained from 14 women inhibited the localized adherence of an enteropathogenic Escherichia coli (EPEC) to HEp-2 cells. Inhibition decreased as lactation continued even when the concentration of sIgA was maintained constant at 1 mg/ml. sIgA responded to a 94-kDa plasmid-encoded outer membrane protein implicated as the EPEC adherence factor. An oligosaccharide-enriched fraction (OEF) from these samples also inhibited the attachment of this EPEC. Inhibition by OEFs decreased as lactation continued because of a general reduction in oligosaccharide content. Localized adherence of six other EPEC was also inhibited by sIgA and OEF, whereas attachment of isolates with diffuse or aggregative adherence was not inhibited by these fractions. Experiments with purified oligosaccharide fractions revealed that EPEC attach to HEp-2 cells through a carbohydrate-mediated mechanism based on the preferential recognition of fucosylated residues in human milk.

Bacterial Adhesion↗

PubMed1980

Effect of sampling on fatty acid composition of human colostrum.

Samples of colostrum were taken from 19 subjects before and after infant feeding, 3 and 5 days post partum. Milk samples were analyzed by argentation and gas chromatography for 30 fatty acids contained in the lipid fraction. Although small differences were observed between day 3 and day 5 samples, the fatty acid composition of the milk did not vary on either of the days between samples taken before and after nursing. It is concluded that a single sample taken any time during the feeding is representative of the entire nursing.

Colostrum↗

PubMed1988

Purification and properties of biologically active chromium complex from bovine colostrum.

A biologically active, low-molecular-weight, chromium-binding substance present in milk (M-LMCr) was isolated from bovine colostrum and purified more than 2000 times by means of ethanol precipitation and successive ion-exchange and Sephadex gel chromatographies. The purified M-LMCr appeared to be an anionic organic Cr compound with a molecular weight of 1500, as determined by gel permeation chromatography. It contained aspartic acid, glutamic acid, glycine and cysteine in a ratio of 5:4:2:1 and no detectable carbohydrate. Although we were unable to detect nicotinic acid, some ultraviolet-absorbing (lambda max 260 nm) chemical structure was shown to be a constituent. Purified M-LMCr stimulated the rates of both [U-14C]glucose oxidation and [3-3H]glucose conversion into lipid in rat epididymal adipocytes at Cr concentrations greater than 1.5 ng/mL in relation to insulin action. This substance appears to have properties similar to those of glucose tolerance factor in yeast and the low-molecular-weight, chromium-binding substance present in mammalian liver. The role of M-LMCr in Cr nutrition and detoxication is discussed.

Amino Acids↗

PubMed1993

Use of the colostrum-deprived piglet to evaluate parenteral feeding formulas.

Several different neonatal and infant piglet models have been invaluable as animal models in nutrition research. A preterm colostrum-deprived piglet model has been developed that is delivered at the desired gestational age, is cared for using the standard of care provided preterm human neonates including procedures for nutritional support and can be studied during pathological conditions induced under controlled conditions. The absolute values for each nutrient requirement would not be expected to be identical for preterm piglets and preterm human neonates. However, the effect of different levels of gestational maturity and superimposed pathophysiologies on nutrient requirements should be similar in the piglet and human neonate.

Animals↗

PubMed1987

Solid-phase assembly of cow colostrum trypsin inhibitor gene.

A gene for cow colostrum trypsin inhibitor (CTI) was constructed from synthetic oligonucleotides using a novel method of solid-phase gene assembly. In the first step an anchor oligonucleotide was covalently bound to the CNBr-activated Sephacryl S-500 support. Next, triads or tetrads of separately annealed oligonucleotides were stepwise hybridized to the immobilized complementary sequence, with washing after each step. In the last step a linearized vector molecule was ligated to the assembled gene. The whole construct was released from the solid support with a restriction enzyme, circularized, and used for transformation, with a high yield of recombinant clones being obtained. The method represents a generally applicable approach to rapid and efficient assembly of extended DNA duplexes.

Acrylic Resins↗

PubMed1984

Purification and characterization of a bovine colostrum low molecular weight cysteine proteinase inhibitor.

Large amounts of cysteine proteinase inhibitors were found in bovine colostrum. One had a molecular weight of 90,000, and the other a molecular weight of 10,500. The concentrations of both these inhibitors were highest the day after parturition, and were about one-tenth as much on day 7. The lower molecular weight inhibitor was purified by acid treatment, ammonium sulfate fractionation, gel filtration on Sephadex G-50, CM-Sephadex chromatography and rechromatography on Sephadex G-50. The purified preparation gave a single band on SDS-polyacrylamide gel electrophoresis. This inhibitor contained one tryptophanyl residue and one cystinyl residue, and did not contain a free thiol group. Values obtained for its isoelectric point (pI) were 10.0 and 10.3. This material strongly inhibited cathepsin B, cathepsin H, and papain. the higher molecular weight inhibitor was partially purified. It had a pI of 4.2 and inhibited papain, cathepsin H, and cathepsin B.

Amino Acids↗

PubMed2004

Antioxidant micronutrient profile (vitamin E, C, A, copper, zinc, iron) of colostrum: association with maternal characteristics.

The study was conducted to investigate the micronutrient profile of human colostrum, and to assess the association of maternal characteristics to the micronutrients. Colostral concentrations of antioxidant vitamins E, C, and A were 21.34 +/- 8.47, 148.92 +/- 43.64, 0.79 +/- 0.42 micromol/l, respectively. The antioxidant minerals copper, zinc, and iron contents were 19.17 +/- 11.73, 63.69 +/- 12.82, 11.44 +/- 1.46 micromol/l, respectively. Maternal characteristics did not have any influence on the colostral micronutrients.

Adolescent↗