• Journal of Veterinary Clinics
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• v.34 no.2
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• pp.87-89
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• 2017

Apo-transferrin is an iron-binding protein that has been reported to have an antimicrobial effect. It is considered a major component of the host defense mechanism as it limits microbial access to iron. This study was performed to investigate whether bovine apo-transferrin would have an inhibitory effect on the growth of S. pseudintermedius, which is one of the most isolated bacteria from dogs, and to compare the antimicrobial efficacy with bovine holo-transferrin. S. pseudintermedius were grown at $37^{\circ}C$ in 96-well culture plates using Muller Hinton broth containing bovine apo-transferrin or bovine holo-transferrin at concentrations ranging from 0.5 or 2.5 to 5.0 mg/ml. The optical densities of the wells were then measured at 570 nm. In this study, the apo-transferrin showed dose-dependent antimicrobial effect against S. pseudintermedius while holo-transferrin did not inhibit the growth of S. pseudintermedius effectively. The results suggest that iron deprivation is an important pathway for inhibiting bacterial growth and bovine apo-transferrin has great antimicrobial effects against S. pseudintermedius.

• Journal of Embryo Transfer
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• v.12 no.3
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• pp.283-291
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• 1997

In vitro development of bovine embryos is affected by many factors such as energy substrates, amino acids, and some growth factors. It has been reported that mRNA of insulin, PDGF and their receptors are detected in cow embryos, and that some chelating agents such as EDTA and transferrin have beneficial role on mouse and bovine embryos. The author hypothesized that insulin, transferrin arid PDGF added to a culture medium increase in vitro development of bovine embryos by chelating toxic substance(s) or increasing cell growth and metabolism. Immature oocytes from slaughtered ovaries of Holstein cows and heifers were matured for 24 hours in a TCM199 containing 10% fetal calf serum, FSH, LH and estradiol with granulosa cells in vitro. Matured oocytes were coincubated with sperm for 30 hours in a modified Tyrode's medium (IVF). Embryos cleaved to 2- to 4-cell at 30 hours after IVF were selected and cultured in a 30-$\mu$l drop of a synthetic oviduct fluid medium (SOFM) containing 0.8% BSA, Minimum Essential Medium essential and non-essential amino acids, and insulin, transferrin or PDGF for 9 days. Supplementation of a SOFM with insulin, and /or transferrin did not increase develop-mental rate to expanding and hatching blastocyst of 2- to 4-cell bovine embryos compared with control. The highest developmental rate to hatching blastocyst was shown when PDGF was added at the concentration of 10 ng /ml among the supplementing doses tested in the present study (p<0.05). Addition of PDGF without insulin to a SOFM could not increase embrye development, but combined addition of PDGF with insulin significantly increased (p<0.05) embryo development to hatching blastocyst (50%) compared with control (38%). In conclusion, insulin and PDGF supplemented to a SOFM may act synergistically and have beneficial effect on in vitro development of 2- to 4-cell bovine embryos matured and fertilized in vitro.

• Journal of Veterinary Clinics
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• v.19 no.1
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• pp.86-91
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• 2002

The inhibitory effect of pooled canine serum on the growth of ten strains of Malassezia pachydermatis in vitro was investigated. Studies were also carried out to observe the effect of different concentrations of unsaturated bovine transferrin on Malassezia growth in vitro. Ten strains of Malassezia pachygedrmatis in normal canine serum (11.1%, 44.4%) were found to significantly inhibit the growth (p<0.0005) not only in a dose dependent but also in a time dependent manner. The same strains of yeast treated with 1, 2, 4 and 8 times the normal value of serum transferrin (3.0 mg/ml, 6.0 mg/ml, 12.0 mg/ml, 24.0 mg/fl), were shown to have significantly lower OD readings (p<0.05) when compared to yeast treated in lower concentrations of transferrin (1.5 mg/ml). The optical density (OD) of the ten strains of yeast were significantly lower (p<0.005) when treated with various concentrations of transferrin than with the saline control except at 72 hours post incubation. These results indicate that serum has inhibitory effects on Malassezia pachydermatis growth in vitro, and transfferin is one of the components that contribute towards this inhibitory role. The inhibitions are dose and time dependent.

• Journal of Dairy Science and Biotechnology
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• v.27 no.1
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• pp.19-28
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• 2009

Lactoferrin was first identified 60 years ago as a "red protein" in bovine milk. Lactoferrin, one of the transferrin family proteins, is an iron-binding glycoprotein found in milk and various mucosal secretions; it is also released from activated neutrophils. Human lactoferrin has a molecular weight of 82.4 kDa and is composed of 702 or 692 amino acid residues. Bovine lactoferrin has a molecular weight of 83.1 kDa and is composed of 689 amino acid residues. Both lactoferrin and transferrin have the ability to bind two $Fe^{3+}$ ions, together with two ${CO_3}^{2-}$ ions with extremely high affinity; these proteins also have the ability to release this iron at low pH levels. The polypeptide chain in lactoferrin is folded into two globular lobes, representing the N-terminal and C-terminal halves. Both lobes have similar folding and 40% sequence identity. This protein is capable of multiple functions as described in various review papers, including antimicrobial, antiviral, antiinflammatory, anticancer, antioxidant, and cell growth-promoting activities. Lactoferrin also exhibits immunomodulating effects and plays an active role in the regulation of myelopoiesis and the inhibition of bacterial translocation.

• Asian-Australasian Journal of Animal Sciences
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• v.14 no.1
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• pp.7-12
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• 2001

Factors Affecting the Survival of Frozen Thawed Bovine In Vitro Produced Blastocysts. The effect of some factors on the post-thaw survival of a total of 240 in vitro produced bovine blastocysts was investigated using logistic regression analysis. The explanatory variables tested were: type of culture medium before freezing (TCM 199 supplemented with BSA, BSAITS (BSA+insulin+transferrin+selenium), ECS (estrous cow serum) with or without BOEC (bovine oviductal epithelial cells), age of the blastocyst (Day 7, Day 8+9), morphological appearance before freezing (distinct=Q1 or indistinct=Q2 inner cell mass) and type of cryoprotectant (glycerol, 1.0 M or ethylene glycol, 1.6 M). The survival after thawing based on the post-thaw quality and the development after co-culture with BOEC for 24 and 48 hours. Day 7 blastocysts had an almost three times better chance of survival than Day 8+9 blastocysts. Q1, Day 8+9 blastocysts had higher odds to survive after 48 hours in culture than Q2 blastocysts (p<0.05). Blastocysts produced in BSAITS medium had the best chances of survival; however, the odds were not always significant. Blastocysts frozen in glycerol had a better post-thaw quality rating than those frozen in ethylene glycol; however, the difference in post-thaw development at culture was not significant. The relationship between post-thaw quality and post-thaw development at culture was significant (p<0.05). The developmental stage and/or age of the embryo and culture medium where development up to blastocyst takes place affect the post-thaw survival of the bovine embryos.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.136-140
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• 2000

A sensitive and quantiative method for the structural analysis of oligosaccharide was established for the glycoform analysis of glconproproteins. Inthis study, n-linked oligosaccharides of human IgG and bovine transferin were analyzed for the evaluation of the methydrate moiety ofthe method. Chrbohydrate moiety of glycoprotein was relased by hydrazinolysis and purified by paper chromatography. The oligosaccharides were labeled with a fluorescent bye, 2-aminobenzamide, for the enhancement of detection sensitivity. sialylated (acidic) oligosaccharides were separated from neutral oligosaccharide by employing a strong anion-exchange column(MonoQ) followed by the treatment with sialidase. Enzymatically desiayated fractions and neutral fractions of oligosaccharides were applied to normal-phase HPLC to resolve the peaks according to glucose unit (GU). The structure of separated molecules was further determined by sequential digestion with exoglycosidases. As a result, disialylated biantennary complextype oligosaccharide was found to be a major sugar chain in bovine transferrin (63%). In human IgG, core fucosylated asialobiantennary complex oligosaccharides were dominant. These results coincided well with reported results.

• The Korean Journal of Physiology and Pharmacology
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• v.14 no.4
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• pp.205-212
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• 2010

Endometritis is one of the primary reasons for reproductive failure. In order to investigate endometritis-associated marker proteins, proteomic analysis was performed on bovine endometrium with endometritis. In bovine endometritis, desmin, $\alpha$-actin-2, heat-shock protein (HSP) 27, peroxiredoxin-6, luteinizing hormone receptor isoform 1, collectin-43 precursor, deoxyribonuclease-I (DNase-I), and MHC class I heavy chain (MHC-Ih) were up-regulated. In contrast, transferrin, interleukin-2 precursor, hemoglobin $\beta$ subunit, and potassium channel tetramerisation domaincontaining 11 (KCTD11) were down-regulated in comparison to normal endometrium. The proteomic results were validated by semiquantitative-PCR and immunoblot analysis. The mRNA levels of desmin, transferrin, $\alpha$-actin-2, HSP27, KCTD11, and MHC-Ih were up-regulated by over 1.5-fold, and showed a pattern similar to their proteomic profiles. Desmin and $\alpha$-actin-2 protein showed positive correlations between proteomic analysis and immunoblot analysis. These results suggest that desmin and $\alpha$-actin-2 may play important roles in endometritis-related function, and could be useful markers for the diagnosis of bovine endometritis.

• Reproductive and Developmental Biology
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• v.30 no.3
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• pp.163-167
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• 2006

The purpose of the present study was to establish culture conditions for the in vitro study of the neonatal piglet Sertoli cell. Isolation for the culture of Sertoli cell was established using collagenase and pancreatin digestion of testicular tissues. The effects of various culture media, fetal bovine serum(FBS), follicular stimulating hormone(FSH), epidermal growth factor(EGF) and insulin-transferrin-sodium selenite(ITS) on growth of neonatal piglet Sertoli cells were investigated. The mitogenic effects of Dulbecco's modified Eagle's medium+Ham's F-12 medium was higher than other media used in this experiment. The addition of 1% FBS in cultures was necessary for attachment of Sertoli cell clusters. However, except FBS and EGF, FSH and ITS did not stimulate Sertoli cell proliferation. When Sertoli cells isolated from neonatal piglets were cultured in Dulbecco's modified Eagle's medium+Ham's F-12 medium supplemented with 1% FBS, FSH EGF and ITS, the yield and plating efficiency of Sertoli cells were largely increased. Confluency of Sertoli cells was reached as early as 4 days of culture. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate and culture of Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro.

• Korean Journal of Food Science and Technology
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• v.29 no.5
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• pp.1067-1070
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• 1997

Imunoglobulins from bovine plasma proteins were isolated by IMAC which $Cu^{2+}$ was chelated on a chelating sepharose fast flow gel. Most plasma proteins were eluted by 1st (0.01 M $Na_2HPO_4$, 0.5 M NaCl, pH 4.0) and 2nd elution buffers (0.01 M imidazol). According to the reverse phase HPLC analysis, it was found that proteins which were eluted by 1st elution buffer were mainly composed of serum albumin, while most IgG and transferrin were eluted by 2nd elution buffer. When protein fractions obtained by 2nd elution buffer was applied to ultra filtration system (molecular weight cut off: 100 kD), IgG was further purified. These results indicate that IMAC is an excellent tool for isolating imunoglobulins from plasma proteins.

• Microbiology and Biotechnology Letters
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• v.17 no.5
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• pp.489-493
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• 1989

The role of each supplement in serum-free medium KM3 for the growth of hybridoma and the production of monoclonal antibody was investigated. Transferrin, ethanolamine and bovine serum albumin were shown to be indispensable for the growth of four kinds of hybridoma tested in this work, especially transferrin for Alps 25-3, and ethanolamine for A4W and KW hybridoma. The addition of $\beta$-mercaptoethanol to the culture medium of HCGK showed a good influence of both the cell growth and the production of monoclonal antibody. Upon the experimental results, we suggested a serum-free medium containing a minimum composition for the culture of hybridoma.