• Journal of Aquaculture
• /
• v.11 no.2
• /
• pp.271-278
• /
• 1998

To clarify characteristics of yolk protein of abalone, yolk protein was purified from the ovarian egg extracts of mature female Haliotis discus hannai by a gel chromatography of sepharose CL-4B. From the results of immuno-electrophoresis and Ouchterlony's diffusion test to male and female sera and ovarian egg extracts using antibodies raised against mature female and male sera and male sera and ovarian egg extrascts, it was identified that the mature female serum had female specific serum protein and its antigenecity shared with ovarian egg extracts. A single type of yolk protein was purified from ovarian egg extracts, and it was composed of two subunits. Their molecular weights were estimated to be approximately 166 KDa and 113 KDa by SDSPAGE. The antiserum against yolk proteins cross-reacted with a mature female specific serum protein and extracts of hepatopancreas of vitellogeing females, but did not reacted with extracts of hepatopancreas of mature male.

• Journal of Nutrition and Health
• /
• v.29 no.6
• /
• pp.569-577
• /
• 1996

This study was performed to determine the effect of maternal protein intake on 1) the concentration of immune substances in milk 2) degree of passive immunity to pups via lactation, and 3) specific antibody production to a specific antigen, $\beta$-lactoglobulin(BLG). 4) the effect of passive immunity that pups received from mother during lactation on the production of antibodies when the pups were challenged to the same antigen. Part of the female rats were immunized with BLG before and during pregnancy. The pregnant rats were placed into either 25% or 10% isolated soy protein diet throughout gestation and lactation. After weaning, pups from each group continued to be fed the same diet. At 18 weeks of age, all the pups were challenged with BLG. Total IgA and IgG, lysozyme, BLG-specific IgA and IgG were measured in dam's serum, dam's milk, and pup's serum. Total IgG, and lysozyme in dam's serum and milk were higher in high protein group. Total IgA and IgG in pup's serum remained higher in high protein group from 5 to 18 weeks of age. BLG-specific antibodies were found in the milk and serum of immunized dams, and in serum of pups born to immunized dams but not in the non-immunized group. BLG-specific IgA and IgG were again higher in high protein group and declined with time. The concentration decreased faster in the low proetein group than in the high protein groups. After immunization the pups with LBG, serum BLG-specific antibodies were not differ between rats born to immunized dams and those born to non-immunized dams. Therefore passive immunity rats received via milk as a pup had no effect on the BLG-specific antibody production later in life. This study shows the importance of protein status of mother and strongly support to the endorsement of breast feeding.

• Korean Journal of Fisheries and Aquatic Sciences
• /
• v.25 no.5
• /
• pp.432-442
• /
• 1992

To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

• Journal of Aquaculture
• /
• v.8 no.1
• /
• pp.1-19
• /
• 1995

In red seabream, Pagrus major the female specific protein in the vitellogenic female serum was identified by Ouchterlony's immunodiffusion test and immunoelectrophoresis. The female specific serum protein might be vitellogenin based on the results of the immunological analysis for the male and vitellogenic female sera and crude egg extracts. Also, it was identified by the immunodiffusion test that the purified yolk protein from ovarian egg extracts has antigenic identities shared with the female specific serum protein. To study the relationship between the maturational stages of gonad and plasma levels of vitellogenin, these were measured from the late resting period (January) to the vitellogenic preiod (April) by the modified enzymeimmunoassay (EIA) using antiserum against yolk protein. The level of plasma vitellogenin began to increase in February (previtellogenesis stage) and continuously increased with the ovarian growth during the vitellogenesis period (March to April). The plasma vitellogenin levels were significantly different between the females and the males in February. Validation for the modified EIA system. was tested .The absorbance curve of serial dilutions of serum from the vitellogenic female was paralleled to the standard curve of yolk protein; $109\pm5.6\%$ recovery was achieved by the modified EIA. And the intraassay coefficients of variation were less than 10% within the concentration ranging from 31.3 ng/ml to 1,000 ng/ml. These findings suggest that the sex determination in adult red seabreams could be possible by using the modified EIA as early as in February.

• Animal cells and systems
• /
• v.15 no.2
• /
• pp.147-154
• /
• 2011

Animal cell cultures generally require a nutrient-rich medium supplemented with animal serum. Adult bovine serum contains a variety of nutrients including inorganic minerals, vitamins, salts, proteins and lipids as well as growth factors that promote animal cell growth. To evaluate the potential use of gender-specific bovine serum (GSBS) for cell culture, the biochemical properties of male serum (MS), female serum (FS) and castrated-male serum (CMS) were investigated. Overall, the chemical profile of GSBS was similar to that of bovine references except for glucose, creatine kinase, lactate dehydrogenase and potassium. FS showed elevated total protein and sodium concentrations compared to MS and CMS. Proteins present in MS, FS and CMS but absent in fetal bovine serum (FBS) were selected by two-dimensional gel electrophoresis and identified by peptide mass fingerprinting. Some of the identified proteins are known to be involved in immune responses and the others have unknown physiological roles. Moreover, it was found that some proteins such as alpha-2-macroglobulin appeared to be gender-specific with higher contents in FS. Insulin and testosterone was significantly higher in MS, and $17{\beta}$-estradiol and estrone were higher in FS, as compared to the other sera. Taken together, the results indicate that each GSBS has a different ratio of components. Differences in serum constituents may affect cell cultures in a different manner and could be beneficial, depending on the specific aim of cell cultures.

• Journal of Aquaculture
• /
• v.9 no.1
• /
• pp.83-92
• /
• 1996

This study was conducted to compare the immunochemical properties of female-specific serum proteins (vitellogenin, VTG) and egg yolk proteins in female fusilier, Caesio diagramma. VTG of fusilier was identified and characterized by using immunochemical analysis. Two types of VTG (VTG1 and VTG2) reacted clearly with antiserum against egg proteins, were confirmed in the serum of mature female. The results of sephacryl S-300 showed that the molecular weights of VTG1 and VTG2 were 560,000 and 410,000, respectively. Yolk proteins, E2 and E3, were isolated from egg extracts, and molecular weights of them were estimated 410,000 and 170,000, respectively. The treatment of $17\beta$-estradiol ($E_2$) to males has induced the synthesis of VTG of which immunological characteristics seems to be similar to the yolk proteins. The results suggest that VTG can be synthesized in the liver by the action of $E_2$ stimulation, and incorporated into the oocytes through the blood circulation. The level of serum $E_2$ was moderately high throughout the spawning period of June. The level of serum VTG was also sustained at high in May and June. The concentration changes of serum $E_2$ and VTG were corelated to the ovarian development in female fusilier. The results indicated that $E_2$ may have some important roles for the vitellogenesis in female fusilier. Also) the VTG can be a precursor protein of yolk not only because it could be synthesized in the liver then incorporated into the oocytes but also because an egg yolk protein had the similiar molecular weights and antigenecity with VTG.

• Journal of Nutrition and Health
• /
• v.29 no.9
• /
• pp.971-980
• /
• 1996

The soluble transferrin receptor(TfR) in human serum has been shown recently to be a truncated form of intact membrane bound receptor containing most of the extracellular domain. We purfied the transferin-free TfR from human serum by immounoaffinity chromatography which produced the single protein identity in high resolution gel chormatography. The monoclonal antibodies(MAb) against purifed serum TfR were produced by fusion of spleen cells o fimmunized Balb/c mice and SP2 cells. Ten hybrids producing MAb specific for serum TfR were identifed and determine their iostypes. A immunoraddiometric assay (IRMA) for serum TfR was established using two monoclonal IgG1 antibodies as the coating and indicator antibodies on the bosis of their suitability in sandwich IRMA of serum TfR. The mean serum TfR levels in the 15 normal male, 15 normal female, and 19 iron-deficient subjects were 5.4$\pm$0.98, 4.6$\pm$0/76, and 18.0$\pm$12.8mg/1, respectively, and the difference in mean values between normal and iron deficient subjects was significant(p=0.0005). There existed the inverse logarithmic relationship(r=-0.9336, p<0.0001) between the serum TfR and ferritin levels.

• Journal of Nutrition and Health
• /
• v.2 no.4
• /
• pp.167-172
• /
• 1969

Total amount of Calories and protein intakes of fishing villagers show higher level than those of farming and mountain villagers, however, the proportion of animal protein (meat protein) in total amount of protein intakes of mountain villagers is much lesser than those of other two villagers. Blood specific gravity, serum protein content and serum tryptophan content were low in inhabitants of mountainous area than those of other two villagers, and in the serum protein fractions, the Albumin Globulin ratio (A/G ratio) show also lower value in inhabitants of mountainous area than those of other two villagers. It is interesting result that serum tryptophan content (serum protein tryptophan) presented a significant positive correlation with that of serum gamma globulin and a significant negative correlation with that of serum albumin. The fact of farming villagers under twenty year old female showing plasma vitamine deficiency phenomenon, however, the plasma carotene value show higher level indicates that the most of the plasma carotene being hardly transfered to the plasma vitamine A in blood. The thiamine value in urine of mountain villagers show higher level than others, it indicates that there is correlation between their poor protein intakes and amount of thiamine ia urine. And the data obtained in the present study could be established by the result of animal experiment reported by Koyanagi et al. and the result of previous paper of auther.

• Korean Journal of Community Nutrition
• /
• v.10 no.3
• /
• pp.264-270
• /
• 2005

Breastfeeding has been known as the best feeding practice to prevent allergies including atopic dermatitis (AD) However, the benefit on the prevention of allergic disease is still controversial. The objectives of this study were to examine the rate of sensitization to the protein of eggs, cow's milk and soy in exclusively breastfed infants and to evaluate antigen-antibody reaction between breast milk and serum of AD infant. Data on feeding and food hypersensitivity were obtained for 62 AD infants (32 male, 30 female) aged < 6 month who had visited Samsung Medical Center from September 2001 to May 2003. Food hypersensitivity was determined by measuring specific IgE to egg, cow's milk and soy. Specific IgE levels > 0.7 kU/L by CAP assay (Pharmacia, Uppsala, Sweden) were considered positive. The rates of sensitization in breastfed infants were $41.9\%$ (26/62) to egg, $30.6\%$ (19/62) to milk and $18.0\%$ (11/62) to soy. Immunoblotting analyses were performed using breast milk with the matched serum of seven AD infants (4 male/3 female). Binding patterns of AD infant's IgE to breast milk extract showed visible specific band for immunoglobulin, especially in case of a lactating mother who did not completely restricted ingestion of egg, milk and soy. These results indicate that sensitization to food allergen develops via breast milk feeding. Breast milk feeding should be recommended in infants at risk of developing allergic disease, but maternal intake of highly allergenic food might be restricted for prevention and treatment of food allergy among the babies with AD.

• Asian-Australasian Journal of Animal Sciences
• /
• v.17 no.6
• /
• pp.743-749
• /
• 2004

The gene encoding human serum albumin (HSA) was cloned from human liver cDNA library by PCR. The HSA cDNA in size of 2,176 bp, including 1,830 bp of open reading frame, was cloned into the plasmid carried with the 5'flanking sequence of goat $\beta$-casein gene (-4,044 to +2,025 bp) to get a tissue specific expression vector in mammary gland named pGB562/HSA (12.5 kb). A 9.6 kb DNA fragment in which the sequence is in order of goat $\beta$-casein gene regulatory sequence, HSA cDNA and SV40 polyadenylation signals was isolated from the pGB562/HSA by SacI and DraIII cutting, and used to microinject into the pronuclei of mouse fertilized eggs to produce transgenic mice. Three transgenic mice (2 female and 1 male) were identified by PCR and dot Southern blot analysis. The copy numbers of integrated transgene were more than 10 copies in line #21 and #26 as well as over 50 copies in line #31 of transgenic mice. HSA protein collected from the milk of lactating transgenic mice was confirmed by immuno-detection of Western and slot blot. The concentrations of HSA in the milk were from 0.05 to 0.4 mg/ml. An obvious antigen and antibody conjugate could be observed in immunohistochemical stain of mammary gland tissue from lactating day 11 of HSA transgenic mice. The transmission of transgene and its expression was recognized according to the results of RT-PCR and sequences analyses of their progeny.