Kei-ichi Shimazaki

Structural determination of three neutral oligosaccharides in bovine (Holstein-Friesian) colostrum, including the novel trisaccharide; GalNAcαl-3Galβ1-4Glc

Two trisaccharides, and a pentasaccharide were obtained from bovine colostrum. Their chemical structures were determined by using methylation and 13C-NMR analyses as follows: GalNac alpha 1-3Gal beta 1-4Glc, Gal alpha-1-3Gal beta 1-4Glc, GaL beta 1-3[Gal beta 1-4GlcNAc beta 1-6]Gal beta 1-4Glc. GalNAc alpha 1-3Gal beta 1-4Glc, which was identified in this study, is a novel oligosaccharide from natural sources. Gal alpha 1-3Gal beta 1-4Glc and Gal beta 1-3[Gal beta 1-4GlcNAc beta 1-6]Gal beta 1-4Glc (lacto-N-novopentaose) have been already found in ovine colostrum, and in horse colostrum and marsupial milk, respectively.

Analysis of human and bovine milk lactoferrins by Rotofor and chromatofocusing.

1. Isoelectric points of human and bovine lactoferrins were evaluated by Rotofor and chromatofocusing analysis. 2. By Rotofor, the isoelectric value of human lactoferrin fraction was determined at 8.7 and that of bovine lactoferrin at 8.8. 3. By chromatofocusing analysis, human and bovine lactoferrins showed different elution patterns. Human lactoferrin was eluted at pH 6.8-8 and bovine lactoferrin eluted at pH 8.2-8.9.

Immunohistochemical localization of lactoferrin in bovine exocrine glands

The localization of lactoferrin (LF) was studied in bovine exocrine glands by the peroxidase-antiperoxidase method. In general, LF immunoreactivity was demonstrated in most of the serous demilunes of the mixed glands and in some but not all acinar cells of the serous glands. LF was not detected in the acinar cells of the mucous glands. LF immunoreactive cells were seen in the nasal and tracheal glands. The lacrimal and mammary glands were stained intensely, while the sweat glands were not stained. In the major and minor salivary glands, LF immunoreactivity was demonstrated in the serous demilunes and acini of the mixed glands and in some serous cells of the pure serous parotid gland. LF immunoreactivity was not seen in the gastrointestinal tract and pancreas, but the serous gland in the lamina propria of the bile duct was stained positively. In the male reproductive tracts, the prostate and bulbourethral glands were stained positively, but the vesicular gland was negative. In the female reproductive system, the uterine and Bartholin glands, the epithelial cells of the cervix and ampulla of the uterine tube reacted positively. However, the lesser vestibular gland did not react for LF. The present findings, that LF was demonstrated in the exocrine glands located mainly in the gateways of the digestive, respiratory and reproductive systems, may morphologically support the antibacterial role of LF.

Different profiles of induced cotton effects of human and bovine lactoferrin by cibacron blue F3GA binding.

1. The properties of complex formation of lactoferrin with Cibacron Blue F3GA dye have been studied by circular dichroic spectral analyses. The Cotton effects were induced by the interaction of lactoferrin with the dye and occurred in the wavelength range from 300 to 450 nm. 2. The patterns of changes in circular dichroic spectra of lactoferrin induced by the dye were different in bovine and human lactoferrin. 3. Iron ions bound to lactoferrin affected the profiles of induced Cotton effects in human lactoferrin but not in bovine lactoferrin. 4. While the dye bound co-operatively to human apo-lactoferrin, such co-operativeness was not observed in iron-saturated human lactoferrin.