It has long been known that lactoferrin prevents human beings from infection of virus. To prove this activity of lactoferrin, we evaluated the activities of different lactoferrins to an isolate human rotavirus K-21. Bovine lactoferrin inhibited infection of K-21 to MA-104 cell at the concentration of 25.9 microM whereas bovine hydrolysed lactoferrin prevented rotavirus infection at 103.8 microM. However human lactoferrin prevented infection of K-21 at the concentration of 217.5 microM. These data suggested that lactoferrin activity may be unaffected by the intestinal digestive enzymes and bovine lactoferrin is more active than human lactoferrin with respect to prevention of rotavirus infection.
Humans* ; Lactoferrin* ; Rotavirus Infections ; Rotavirus*

It has long been known that lactoferrin prevents human beings from infection of virus. To prove this activity of lactoferrin, we evaluated the activities of different lactoferrins to an isolate human rotavirus K-21. Bovine lactoferrin inhibited infection of K-21 to MA-104 cell at the concentration of 25.9 microM whereas bovine hydrolysed lactoferrin prevented rotavirus infection at 103.8 microM. However human lactoferrin prevented infection of K-21 at the concentration of 217.5 microM. These data suggested that lactoferrin activity may be unaffected by the intestinal digestive enzymes and bovine lactoferrin is more active than human lactoferrin with respect to prevention of rotavirus infection.
Humans* ; Lactoferrin* ; Rotavirus Infections ; Rotavirus*

PURPOSE: To evaluate the changes of tear osmolarity and protein after silicone punctal plug insertion in dry eye. METHODS: We collected tear from 21 dry eyes (11 males and 10 females) with porous polyester rod(Transorb(R)) before silicone punctal occlusion and 2 weeks after the procedure. Tear osmolarity was measured by Fiske 2400 Osmometer(R). Total tear protein concentration was analyzed by Bradford's assay. The concentration of human serum albumin (HSA), lactoferrin, lysozyme were measured by Bio-Rad Gel Doc 2000(R). RESULTS: The tear osmolarity was changed from 364.48+/41.39 mOsm/L to 327.48+/-27.24 mOsm/L after the punctal occlusion (P<0.05). There were no changes of total protein concentration, Human serum albumin, lactoferrin, but lysozyme was increased statistically significantly (P<0.05). CONCLUSION: Silicone punctal occulusion decreases tear osmolarity in dry eye. This decrease is associated with a decrease in ocular surface disease due to high tear osmolarity. Only lysozyme except HSA and lactoferrin was increased significantly. Silicone punctal plug is an effective procedure for dry eye patients, but further investigation of the effects on tear proteins is needed.
Humans ; Lactoferrin ; Male ; Muramidase ; Osmolar Concentration* ; Polyesters ; Serum Albumin ; Silicones*

During bone remodeling, there is requirement of differentiation of osteoblastic cells. Previously, we identified proteins differentially expressed in soft tissue during bone healing. Of these proteins, we focused the effect of LTF on differentiation of osteoblast. In order to analyze the osteogenic ability of LTF, we treated conditioned media collected from human LTF-stably transfected HEK293T cells into osteoblastic MC3T3-E1. The results showed that the activity and expression of alkaline phosphatase were increased in MC3T3-E1 cells treated with conditioned media containing LTF in dose- and time-dependent manner. At the same time, we observed the significant increase of the expression of osteoblastic genes, such as ALP, BSP, COL1A1, and OCN, and along with matrix mineralization genes, such as DMP1 and DMP2, in LTF conditioned media-treated groups. Moreover, the result of treating recombinant human LTF directly into osteoblastic MC3T3-E1 showed the same pattern of treating conditioned media containing LTF. Our study demonstrated that LTF constitutively enhances osteoblastic differentiation via induction of osteoblastic genes and activation of matrix mineralization in MC3T3-E1 cells.
Alkaline Phosphatase ; Bone Remodeling ; Culture Media, Conditioned ; Humans ; Lactoferrin* ; Osteoblasts*

Lactoferrin (Lf) is one of the food protein belonged to the innate immune system. Apart from its main biological function of binding and transport of iron ions, lactoferrin also has many other functions and properties such as antibacterial, antiviral, antiparasitic, catalytic, anti-cancer, anti-allergic and radioprotecting. Lf is usually used as additives of food and cosmetics. The research of lactoferrin has been increasingly reported, and the application of lactoferrin as a drug carrier has drawn extensive attention over the recent year. In this paper, researches of lactoferrin as drug carriers are classified and summarized in brain targeting, liver tumor targeting, lung tumor targeting and oral delivery systems according to their different characteristics.
Administration, Oral ; Brain ; Drug Carriers ; Humans ; Lactoferrin ; chemistry ; Neoplasms

The proportion of tear proteins is different markedly in various ocular diseases, so its measurement may be of considerable diagnostic value in certain ocular conditions. We studied prospectively proportion of tear proteins in 50 normal adult Koreans. 25 male and 25 female, by using protein electrophoresis. Four fractions were demonstrated by protein electrophoresis using cellulose acetate membrane: Fraction 1 represents as tear-specific prealbumin(TSP1) and albumin, fraction 2 as other tear-specific prealbumin(TSP2), fraction 3 as lactotransferrin and immunoglobulin, and fraction 4 as lysozyme. In this study, the proportion of tear proteins in fraction 1 to 4 were 33.4%, 1.1%, 44.2%, and 21.2% respectively. With age, the proportion of fraction 4 decreased, and its decrease was not statistically significant difference. There was no statistically significant difference between age groups in fractions 1,2 or 3. Difference of proportion of tear proteins between the sex was not statistically significant in any of the fractions. Further studies on the tear proteins may be helpful in diagnosing some ocular conditions.
Adult ; Cellulose ; Electrophoresis* ; Female ; Humans* ; Immunoglobulins ; Lactoferrin ; Male ; Membranes ; Muramidase ; Prospective Studies ; Tears*

B cell-activating factor belonging to the TNF family (BAFF) is primarily expressed by macrophages and stimulates B cell proliferation, differentiation, survival, and Ig production. In this study, we explored the effect of lactoferrin (LF) on BAFF expression by murine macrophages. We determined the level of BAFF expression at the transcriptional and protein levels using RT-PCR and ELISA, respectively. LF markedly enhanced BAFF expression in mouse macrophages at both the transcriptional and protein levels. Overexpression of Smad3/4 further increased LF-induced BAFF transcription while DN-Smad3 abolished the LF-induced BAFF expression. These results demonstrate that LF can enhance BAFF expression through Smad3/4 pathway.
Animals ; Cell Proliferation ; Enzyme-Linked Immunosorbent Assay ; Humans ; Lactoferrin ; Macrophages ; Mice ; Transforming Growth Factor beta1

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

The supramolecular inclusion properties of beta-cyclodextrin (beta-CD) and resveratrol (Res) were investigated using drug-protein interaction spectroscopy method. The differences between the results of interaction spectroscopy method and the results of classical method were compared. The total energy of the stable inclusion of cyclodextrin-resveratrol was calculated by Gaussian theory calculation. The stable inclusions in the process of interaction between resveratrol/inclusion complex and bovine lactgoferrin (BLF) were studied by molecular modeling. The results showed that the interaction spectroscopy method could explain the property of the inclusion in a more sensitive manner, it also interpreted the conveying mechanism of BLF binding with inclusion complex. The molecular modeling result showed consistent results with Gaussian theory calculation; both of the two methods obtained the stable configuration of beta-CD-Res inclusion. The relevant result provided an experimental consequence for the pharmacology research of beta-cyclodextrin-resveratrol inclusion complex as well as offering a new reference to the future research of supramolecular inclusion compound.
Animals ; Cattle ; Lactoferrin ; chemistry ; Magnetic Resonance Spectroscopy ; Models, Molecular ; Stilbenes ; chemistry ; beta-Cyclodextrins ; chemistry

BACKGROUNDTo evaluate expression of human lactoferrin gene by high-density fermentation in recombinant Pichia pastoris on the premise of maintaining its biological activities.

METHODSThe neutrophil was isolated from human peripheral blood and its total RNA was prepared. Full-length cDNA of human lactoferrin gene was then obtained by RT-PCR, cloned into expression vector pPIC 3.5 K and transformed into Pichia pastoris strain KM71. With two-layer filter method, the transformants with high-productivity of human lactoferrin were screened out into fed-batch high-density fermentation. And later, the physical, chemical and biological activities of fermentation product were detected preliminarily.

RESULTSThe strain p3.5-k-7 with better productivity of human lactoferrin was screened out into fed-batch high-density fermentation. The fermentation lasted nearly for nine days, with A-600 of culture once above 260 and the highest productivity of human lactoferrin being 115 mg/L, 7.67 times the amount of that in shake flask cultivation.

CONCLUSIONThe authors successfully realized high-density fermentation expression of human lactoferrin gene in recombinant Pichia pastoris.

Cloning, Molecular ; Fermentation ; Humans ; Lactoferrin ; biosynthesis ; genetics ; Pichia ; genetics ; metabolism ; Recombinant Proteins ; biosynthesis