• Journal of Veterinary Clinics
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• v.24 no.3
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• pp.305-307
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• 2007

Lactoferrin-binding proteins (LBP) has not been well characterized in Streptococcus uberis isolated from milk of bovine mastitis and to date this protein is considered to be an important virulence factor in Streptococcal mastitis. To determine the more efficient extraction method of LBP from four S. uberis strains, we used two different extraction methods (mutanolysin and sodium dodecyl sulfate) in this study. Bacterial proteins extracted were electrophoresed by 10% polyacrylamide gels in the presence of sodium deodecyl sulfate and gels were transferred onto nitrocellulose membrane. Rabbit anti-bovine lactoferrin antibody and HRP-conjugated donkey anti-rabbit IgG antibody were used to detect LBP. This Western blotting analysis demonstrates that extraction method with SDS extracted 110 kDa and 112 kDa LBPs more efficiently compared to the mutanolysin extraction method.

• Korean Journal of Agricultural Science
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• v.8 no.2
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• pp.231-237
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• 1981

In order to obtain the basical data for utilization of protein in hazel nut, protein was extracted from defatted hazel nut meal with salt solutions and alkaline solutions, and precipitated by adjusting pH of extract to 5.5 or addition of organic solvents. Amino acid composition of the isolated protein and defatted hazel nut meal were analyzed, protein isolates were identified by polyacrylamide gel electrophoresis. The results summarized were as follows. 1. Defatted hazel nut showed highly nutritional value as the content of 'protein was 53.6%. 2. Extractabilities of salt-soluble protein treated with 2.5M $MgCl_2$, and 1M NaCl(pH 11.0) were 53.0%, 31.5%, respectively 3. Protein in hazel nut were contained 53% of salt-soluble globulin, 14% of water-soluble albumin, 29.5% of glutelin based on solubility. 4. At pH 5.5, 85% of the extracted protein was precipitated, and about 90% of the extracted protein was separated by addition of organic solvents such as acetone and ethanol at 60-70% concentration. 5. Proteins extracted from defatted hazel nut with water and 0.027N NaOH showed 3 and 6 hands by polyacrylamide gel electrophoresis, respectively. 6. Amino acids of defatted hazel nut and protein isolate were chiefly composed of glutamic acid, arginine and aspartic acid.

• Journal of the Korean Society of Food Science and Nutrition
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• v.35 no.5
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• pp.594-599
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• 2006

In our previous report, we indicated that not only BSA but also BGG played an important role in the allergenicity of beef. In this study, the effect of heat or high-pressure treatments to beef extract on the SDS-PAGE patterns was examined. The antigenicity of each treated samples was also investigated by Western blots assay with the sera of BGG-positive beef allergic patients. The BGG band and its antigenicity slightly disappeared but not generally in $100^{\circ}C$ group, indicating $100^{\circ}C$ treatment is not sufficient to totally eliminate the antigenicity of beef allergens. Compared with BGG band, BSA band significantly disappeared in SDS-PAGE with $100^{\circ}C$ treatment, indicating BSA is more heat- sensitive than BGG. When the beef extract was heated at $120^{\circ}C$, not only BSA but also BGG bands was largely disappeared in both SDS-PAGE and Western blots. High pressure (HP) treatment even at 600 MPa did not affect SDS-PAGE and Western blots pattern of BSA. On the contrary, BGG treated with HP showed visible changes in SDS-PAGE. 600 MPa treatment significantly reduced the antigencity. Interestingly, these behaviors of BGG were not found in the same experiments with pure BGG treated with HP. From these results, it was speculated that some kinds of proteolytic enzymes in beef extracts were involved in the BGG molecular degradation by HP treatment. The aging experiments of beef extracts treated with HP supported this hypothesis. Further studies are needed to clarify the function and working mechanism of enzymes associated with BGG degradation in beef extracts by HP treatment.

• Journal of Food Hygiene and Safety
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• v.32 no.1
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• pp.75-81
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• 2017

For people who have a food allergy the only way to manage the allergy is to avoid the food allergen. The mackerel is one of the major food allergens, but no immunoassay for the rapid and simple detection of mackerel has been reported. The objectives of this study are to develop and characterize monoclonal antibodies (MAbs) specific to mackerel using thermal stable-soluble proteins (TSSP) as an immunogen and to characterize the MAbs by indirect enzyme-linked immunosorbent assay (iELISA). The mice immunized with mackerel TSSP and showing high titer were used for cell fusion and cloning. The characterization of MAbs produced from hybridoma cells obtained was confirmed by indirect ELISA and western blot. Four MAbs were confirmed to be specific to mackerel without cross-reaction to other marine products and livestock products in the both methods. The iELISA and western blot based on the MAbs can sensitively detect 1% mackerel protein in other marine products. These results support that immunochemical methods based on the MAb produced could be used as rapid means to detect low levels of mackerel and to identify mackerel adulterated in food.

• Journal of the Society of Cosmetic Scientists of Korea
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• v.32 no.4 s.59
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• pp.241-247
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• 2006

This work was carried out to investigate the anti-inflammatory effects of Paulownia coreana Uyeki on abirritant, atopy and acne skin. Paulownia coreana Uyeki has been used as a traditional medicine having anti-febrile, anti-inflammation effect in Korea, Paulownia coreana Uyeki loaves were extracted with 70% EtOH. Its superoxide anion radical scavenging activity and inhibitory effect on LPS-induced NO production were examined. The extract inhibitied the generation of NO and $PGE_2$ induced by LPS in the macrophage cell line (Raw 264.7). Consistent with the inhibitory effects on No and $PGE_2$ generation, the extract inhibited expression of iNOS and COX-2. In further study, it was found that the extract prevented $IkB-{\alpha}$ degradation, as demonstrated by western blot analysis of $IkB-{\alpha}$ protein level. However, the extract treatment did not affect cell viability at $100{\mu}g/mL$ concentration in both human skin fibroblast and Raw 264.7 cells in vitro. Thus, the present study suggests that Paulownia coreana Uyeki leaves extract have significant anti-inflammatory activity and potential as an anti-irritation material.

• Journal of the Korean Society of Food Science and Nutrition
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• v.43 no.9
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• pp.1462-1466
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• 2014

In this study, an attempt was made to produce oat protein concentrates from defatted oat groat by alkali extraction. Independent variables formulated by D-optimal design were NaOH concentration (X1, 0.005~0.06 N) for extraction and precipitation pH (X2, pH 4.0~6.0), and the dependent variable was extraction yield (Y1, %). Experimental results were analyzed by response surface methodology to determine optimized extraction conditions. Extraction yield increased both with an increase in NaOH concentration of the extraction solution and when approaching a precipitation pH of 4.9, and NaOH concentrations were a major influencing parameter. Solubility of oat protein concentrates showed a minimum value (i.e., 0.1%) at pH 5 and increased substantially at pH values in the range of ${\leq}$ pH 3 or ${\geq}$ pH 7, reaching a maximum value at pH 11 (i.e., 76%). Regression equation coincided well with the results of the experiment. Optimized extraction conditions to maximize extraction yield were 0.06 N NaOH (X1) for extraction and pH 4.7 (X2) for precipitation.

• Journal of Life Science
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• v.28 no.11
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• pp.1316-1320
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• 2018

The effects of a green tea extract (GTE) were examined using the MCF-7 human breast cancer cell line. Cell viability assays using 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assays revealed that GTE had a significant cytotoxic effect on MCF-7 cells, depending on the concentration of GTE. Western blotting of p53 and its related proteins, p21/cip1 and CDK2, after GTE treatment revealed that a significant and concentration dependent increase in p53 protein in response to GTE. The levels of p21/cip1 proteins were also increased at low GTE concentrations were significantly increased even at the highest GTE concentrations. However, the level of CDK2 was significantly decreased by treatment with high concentrations of GTE. These results indicate that treatment with GTE increased the p53 level in MCF-7 cells, and this activation of p53 markedly elevated the levels of p21/cip1proteins, which, in turn, inhibited CDK2 expression in the MCF-7 cells. The inhibition of CDK2 expression might then affect cell cycle progression. Subsequent FACS analysis indicated that GTE treatment the gradually increased progression of the MCF-7 to the G1 phase. These results clearly demonstrate that the anti-tumor effect of GTE in MCF-7 cells is regulated by p53 arrest of the MCF-7 cells at the G1 stage of cell cycle.

• Parasites, Hosts and Diseases
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• v.31 no.3
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• pp.269-276
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• 1993

To ascertain that tegument of Pnragonimus westermoni has specific antigenic proteins, the tegumental fraction was isolated from 10-month-old worms by 0.1% digitonin solution, and subjected to SDS-PAGE and immunoblot. Component proteins of tegumental syncytium comprised of 94, 74 (76-66), 62, 54, 44, 42, 38, 28, 26, 25, 24, 17, 15.5 and 13.5 kDa proteins. Of them, the 94, 44 and 42kDa proteins were more specific to tegument, especially the 94 kDa protein was the most prevailing one. In immunoblot, antigens of the 94, 90, 78, 76, 74, 68, 65, 63, 60, 59 and 54 kDa proteins were commonly detected by 7 sera of 10 human paragonimlasis, but none of them reacted with 5 sera of clonorchiasis. In conclusion, the 94 kDa protein was the major tegumental protein, as well as the specific antigen. The 76 and 66 kDa proteins were the minor components of tegument, which were also specific antigens of R westermcni.

• Applied Biological Chemistry
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• v.42 no.1
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• pp.39-44
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• 1999

Protein profiles of beef loin were constructed by comparing two-dimensional gel electrophoresis patterns of the proteins from different growth stages of Hanwoo, Korean cattle. Proteins from the lean muscle of 0, 6, 12 and 24 months old Hanwoo were separated by isoelectric focusing (IEF) on 16 cm tube gels, and further processed second dimensionally by 12% SDS-polyacrylamide gel electrophoresis (PAGE) using $18{\times}20$ cm gel. Proteins with pI values ranging 3.0 to 9.0 and molecular weight of 15 to 100 kDa could be clearly detected on gel by silver staining. Interestingly, many of the proteins significantly increased and decreased during growth happened to be low molecular ones. To isolate the increased proteins, the soluble proteins were obtained from the tissue by 1% Triton X-100 extraction, then, fractionated by 30% and 50% ammonium sulfate. The isolation condition of each particular protein was determined. According to the conditions, two of the increased proteins were isolated, and transferred to PVDF membrane and microsequenced.

• Food Science and Preservation
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• v.20 no.5
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• pp.597-601
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• 2013

In this study, we searched for bioactive compounds from natural resources with a supercritical extract. We selected the extracts of Chrysanthemum zawadskii, Lufa cylindrica, Paeonia lactiflora, Gardenia jasminoides and Scutellaria baicalensis, as natural materials, and evaluated the effects of their skin barrier function. We found that these extracts increased the transactivation activity of the PPAR-responsive element (PPRE) and the anti-oxidation with different priorities, respectively. In addition, these extracts promoted the expression of proteins related to cornified envelope (CE) formation, such as involucrin. From these results, we suggest that natural materials from supercritical extracts will be pertinent candidates for the improvement of the epidermal permeability barrier function.