• Clinical and Experimental Pediatrics
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• v.49 no.10
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• pp.1061-1066
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• 2006

Purpose : The purpose of this study was to research whether measurement of cow's milk specific IgE on the newborn would be helpful in the diagnosis of cow's milk allergy. We tried to find out the relation between cow's milk specific IgE and other allergy diseases by following up cases. Methods : We reviewed clinical features of 87 episodes in infants less than 4 weeks old who were positive in cow's milk specific IgE test. For the study group, history taking, physical examinations, elimination and cow's milk specific IgE tests were carried out. We investigated the connection among cow' milk specific IgE, allergic disease and family history in 40 of 87 patients we could follow up on. Results : The mean age of the study group was $17.2{\pm}5.4days$. The subjects were classified in four groups according into allergens : 87 milk allergy positive patients, 24 casein positive, 38 ${\alpha}$-lactoalbumin positive, and 75 ${\beta}$-lactoglobulin positive. The number of patients who had follow-ups for more than 6 months to was 40(45.9 percent). The patients whose parents had allergic disease numberred 10(25 percent). Fiften patients had allergic diseases, 4 had asthma and 11 atopic dermatitis. According to the follow-up study, there is a significant relation between casein positive patients and allergic disease. But there is no statistical and significant relation between cow's milk specific IgE and a family history of allergic disease. Conclusion : For the newborn babies, elimination tests and cow's milk specific IgE tests can be useful in the diagnosis of IgE-mediated or mixed milk allergies.

• Journal of the Korea Academia-Industrial cooperation Society
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• v.18 no.10
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• pp.551-558
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• 2017

To study and validate tissue-specific promoters and vectors, it is important to develop cell culture systems that retain the tissue and species specificity. Such systems are attractive alternatives to transgenic animal models. This study established a line of porcine mammary gland epithelial cells (PMECs) from a primary culture based on the cellular morphology and mRNA levels of porcine beta-casein (CSN2). The selected PMECs were stained with the cytokeratin antibody, and were shown to express milk protein genes (CSN2, lactoferrin, and whey acidic protein). In addition, to confirm the acini structure of PMEC932-7 in 3D culture, live cells were stained with SYTO-13 dye, which binds to nucleic acid. The acini of these PMECs on matrigel were formed by the aggregation of peripheral cells and featured a hollow lumens. The system was demonstrated by testing the effects of the culture conditions to cell culture including cell density and matrigel methods of the PMECs. These results suggest that PMECs possess the genetic and structural features of mammary epithelial cells.

• Preventive Nutrition and Food Science
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• v.5 no.4
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• pp.200-204
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• 2000

Protein conjugation of curdlan belonging to $\beta$-1, 3-glucan was carried out to improve it surface functionalities. The glucan was mixed with phosvitin, {TEX}$$\alpha$_{s}${/TEX}-casein, lysozyme or ovalbumin, respectively. The mixture was freeze-dried, and he resulting powder was incubated at 6$0^{\circ}C$ and 79% relative humidity for 12 days in order to generate a controlled Maillard reaction between curdlan and proteins. conjugation with unfolding proteins, i.e., phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein, drastically increased the solubility of the glucan, whereas lysozyme and ovalbmin did not. The solubility in water of curdlan was 3.44% for the phosvitin conjugate and 1.09% for the {TEX}$$\alpha$_{s}${/TEX}-casein conjugate. SDS-slab polyacrylamide gel electrophoresis showed that curdlan was solubilized due to covalent binding with phosvitin. Emulsifying properties of curdlan were substantially improved by the conjugation with phosvitin and {TEX}$$\alpha$_{s}${/TEX}-casein. Emulsion stability of the curdlan-phosvitin conjugate was about 2.9 times greater than that of the curdlan-phosvitin mixture.

• Asian-Australasian Journal of Animal Sciences
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• v.5 no.2
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• pp.285-294
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• 1992

Biochemical polymorphisms of sow's milk proteins, $\beta$-casein ($\beta$-CN), $\beta$-lactoglobulin ($\beta$-LG), post-lactoglobulin (post-LG), $\alpha$-lactalbumin ($\alpha$-LA) and X-protein, as genetic markers for major pig breeds (Landrace, Yorkshire, Duroc, Hampshire and cross bred) in Korea were determined by starch gel electrophoresis. Phenotype and gene frequencies at all marker loci were estimated and genetic differences among breed populations were analyzed. Three $\beta$-CN phenotypes (AA, AB and BB) controlled by two codominant alleles (${\beta}-CN^A$ and ${\beta}-CN^B$), four $\beta$-LG phenotypes (AA, AC, $AC^{\pm}$ and CC) controlled by two codominant alleles (${\beta}-LG^A$ and ${\beta}-LG^C$) and ten X-protein phenotypes (AA, BB, CC, DD, AB, AC, AD, BC, BD and CD) controlled by four codominant alleles ($X^A,\;X^B,\;X^C\;and\;X^D$) were identified. In addition, a genetically controlled polymorphism of post-LG was found for the first time in sow's milk protein. Three different phenotypes (AA, AB and BB) were designated $post-LG^A$ and $post-LG^B$. Of the five marker loci examined, $\alpha$-LA locus was observed to lack any individual variation in all breeds studied. All populations were in Hardy-Weinberg equilibrium for all loci. There were marked breed differences for phenotype and gene frequencies in the post-LG and X-protein marker loci. However, there were little differences between breeds in the gene frequencies at the $\beta$-CN and $\beta$-LG marker loci.

• Reproductive and Developmental Biology
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• v.29 no.4
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• pp.235-239
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• 2005

This study was conducted to test whether the transgenic cattle pass the transgene to their progeny through germ cells, and whether the transgene is expressed in the mammary gland of ransgenic cows. Two male ransgenic calves were born from IVF-derived embryos injected with bovine $\beta-casein/human$ lactoferrin fusion gene and then grew up to be reproducible. Semen was collected from a transgenic bull after 18 mon of age and then frozen. Bovine oocytes matured in vitro were fertilized with spermatozoa of the transgenic bull and cultured in $50\;{\mu}L$ drops of CRlaa medium supplemented with 3 mg/mL BSA. After 48 h of culture, cleaved embryos were determined for the presence of transgenes by DNA polymerase chain reaction (PCR). Proportion of transgene positives among bovine embryos fertilized with sperm of the transgenic bull was $20.9\%$ (28/134). One of transgenic bulls did not produce transgenic sperm. Out of 34 calves produced from recipient heifers inseminated with semen of the other bull, 3 $(8.8\%)$ were transgenic animals (2 females and 1 male). Thus, one transgenic bull showed a low transmission frequency below Mendelian levels in both the IVF-derived embryos and his progeny. It was demonstrated by Southern blot that copy numbers of the transgene in the transgenic progeny enhanced about 1.8 times as compared to those of the founder bull The results demonstrate that the transgenic bull carrying human lactoferrin gene could pass his transgene to the progeny through germ cells, although he is a germ-line mosaic.

• Korean Journal of Food Science and Technology
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• v.17 no.5
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• pp.389-398
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• 1985

To shorten the processing of cheese slurry, four different slurries, ie, Control, Cheddar 1 and 2, and Italian-type that were made of Na-caseinates, cream, trace elements, lactic culture, and enzymes were fermented at $30^{\circ}C$ for 7days with daily stirring. PH, titratable acidity, soluble nitrogen, viable cell count, active SH groups, total volatile fatty acid, free fatty acid, electrophoretic patterns of degraded caseins, and viscosity were analyzed to investigate physicochemical properties of fermented slurries. Acid production was accelerated in the cheese slurries with protease than that without the enzyme and PH of the former was decreased after three days of fermentation to 4.90. The Change of titratable acidity agreed to PH patterns. Soluble nitrogen of the Control slurry was increased slowly for four days and then rapidly to 40% of total nitrogen while those containing protease to 70%. The protease of lactic cultures used (Streptococcus lactis and Streptococcus cremoris) broke down as-casein more rapidly than $\beta$-casein and most proteins were degraded to peptides and amino acids after three days of fermentation. Total volatile fatty acids were increased by added lipase and free fatty acids composition analyzed by GLC in cheddar slurry with 0.00001% lipase was similar to that of commercial cheddar cheese, while that in Italian-type slurry was a half of that in commercial Italian cheese. Active SH groups were increased in the cheese slurries with glutathione from fourth day of fermentation. The viscosity of slurries decreased very rapidly by addition of protease.

• Journal of the Korean Society of Food Science and Nutrition
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• v.22 no.2
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• pp.127-131
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• 1993

To evaluate an effect of dietary protein on lipoprotein profile serum of carbon tetrachloride-treated rats, carbon tetrachloride (50% in olive oil) was twice given at 0.1ml/100g body weight at intervals of 24hours to the male rats and then the degree of liver damage in carbon tetrachloride-treated animals fed a low protein diet was compared with that fed a high protein diet. The increasing rate of liver weight/body weight and the serum levels of alanine aminotransferase in carbon tetrachloride-treated rats to the control group were higher in rats fed high protein diet than those fed low protein diet. In the serum levels of lipid (total lipid, total cholesterol and triglyceride) remarkable differences were not found between low protein diet group and high protein diet group. But these serum lipids in carbon tetrachloride-treated rats were decreased and the decreasing rate of serum lipids to control group were higher in carbon tetrachloride-treated rats fed high protein diet than those fed low protein diet. Under the animal model as identified by the present data herein, serum pre $\beta$-lipoprotein and $\alpha$-lipoprotein fractions were decreased in carbon tetrachloride-treated rats, but the serum levels of $\beta$-lipoprotein were rather increased in the both group by the injection of carbon tetrachloride. Especially, the decreasing rate of $\alpha$-lipoprotein fraction was higher in $CCl_4$-treated rats fed a high protein diet than those fed a low protein diet to its control group and the increasing rate of serum $\beta$-lipoprotein fraction was also higher in $CCl_4$-treated rats fed high protein diet than those fed low protein diet.

• Korean Journal of Food Science and Technology
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• v.34 no.5
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• pp.885-888
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• 2002

Prevalence of food allergic disease was examined by identifying the most common foods implicated in allergic reactions in Korea. Patients were subjected to test determining the amount of specific IgE antibody in serum against food allergens by CAP system. A total 9054 CAP analyses on egg white, egg yolk, cow milk, ${\alpha}-lactalbumin,\;{\beta}-lactoglobulin$, casein, wheat, rice, buckwheat, soybean, peach, crab, shrimp, pork, beef, chicken, tuna, salmon, mackerel, and food mix were undertaken. The results were considered to be positive when CAP value was same and/or greater than +2 (0.7 U/mL). Positive results of CAP analyses were 11.3% (1022/9054 cases), consisting of 336 on egg white, 266 on cow milk, 95 on egg yolk, 76 on soybean, 69 on ${\alpha}-lactalbumin$, 61 on casein, 58 on ${\beta}-lactoglobulin$, 39 on buckwheat, 12 on wheat, 3 on beef, 2 on crab, and 1 on rice, shrimp, pork, chicken or mackerel, and 0 on peach, tuna or food mix. Egg, cow milk, soybean, buckwheat, and wheat were identified as the most common allergic foods in Korea, showing an average of two different food sources for allergy per patient.

• Korean Journal of Food Science and Technology
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• v.32 no.3
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• pp.564-569
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• 2000

An enzyme-linked immunosorbent assay(ELISA) was developed for the detection of milk proteins in processed foods. The ${\alpha}_{s1}-casein({\alpha}_{s1}-CN)$, a heat stable major milk protein, was immunized into rabbits to produce specific antibodies. When competitive indirect ELISA(ciELISA) using $anti-{\alpha}_{s1}-CN$ antibodies was established, its detection limit was $0.1\;{\mu}g/mL$. The reactivities of the specific antibodies toward ${\alpha}_{s1}-CN$, skim milk, ${\beta}-CN$ and whey protein isolate(WPI) were 100, 37, 0.14 and 0.04%, respectively, as determined by ciELISA. However $anti-{\alpha}_{s1}-CN$ antibodies did not have any reactivity to other milk proteins such as ${\beta}-lactoglobulin,\;{\alpha}-lactalbumin$, bovine serum albumin, and isolated soy protein. When sandwich ELISA was established, its detection limit was $0.01\;{\mu}g/mL$ which was 10 times more sensitive than that of ciELISA. In the spike test which was performed by adding 1-10% of whole CN to market milk, mean assay recovery as determined by sandwich ELISA was 94.8%(CV, 8.2%). Food stuffs and dairy products were assayed by sandwich ELISA to show 29, 0.13, 0.25, and 6.9% of whole CN in skim milk powder, WPI, semi-solid yoghurt, and processed cheese, respectively.

• Journal of Animal Science and Technology
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• v.64 no.3
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• pp.481-499
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• 2022

This study aims to determine the effects of D-methionine (D-Met) isomer and the methionine precursor 2-hydroxy-4-methylthiobutanoic acid i (HMBi) supplementation on milk protein synthesis on immortalized bovine mammary epithelial cell (MAC-T). MAC-T cells were seeded using 10-cm dishes and cultured in Dulbecco's modified Eagle's medium/F12 (DMEM/F12) basic medium. The basic medium of DMEM/F12 was replaced with the lactogenic DMEM/ F12 differentiation medium when 90% of MAC-T cells reached confluency. The best dosage at 0.6 mM of D-Met and HMBi and incubation time at 72 h were used uniformly for all treatments. Each treatment was replicated six times wherein treatments were randomly assigned in a 6-well plate. Cell, medium, and total protein were determined using a bicinchoninic acid protein assay kit. Genes, proteomics and metabolomics analyses were also done to determine the mechanism of the milk protein synthesis pathway. Data were analyzed by two-way analysis of variance (ANOVA) with supplement type and plate as fixed effects. The least significant difference test was used to evaluate the differences among treatments. The HMBi treatment group had the highest beta-casein and S6 kinase beta-1 (S6K1) mRNA gene expression levels. HMBi and D-Met treatments have higher gene expressions compared to the control group. In terms of medium protein content, HMBi had a higher medium protein quantity than the control although not significantly different from the D-Met group. HMBi supplementation stimulated the production of eukaryotic translation initiation factor 3 subunit protein essential for protein translation initiation resulting in higher medium protein synthesis in the HMBi group than in the control group. The protein pathway analysis results showed that the D-Met group stimulated fructose-galactose metabolism, glycolysis pathway, phosphoinositide 3 kinase, and pyruvate metabolism. The HMBi group stimulated the pentose phosphate and glycolysis pathways. Metabolite analysis revealed that the D-Met treatment group increased seven metabolites and decreased uridine monophosphate (UMP) production. HMBi supplementation increased the production of three metabolites and decreased UMP and N-acetyl-L-glutamate production. Taken together, D-Met and HMBi supplementation are effective in stimulating milk protein synthesis in MAC-T cells by genes, proteins, and metabolites stimulation linked to milk protein synthesis.