• Journal of Veterinary Science
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• v.23 no.4
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• pp.55.1-55.12
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• 2022

Background: ASF was first reported in Kenya in 1910 in 1921. In China, ASF spread to 31 provinces including Henan and Jiangsu within six months after it was first reported on August 3, 2018. The epidemic almost affected the whole China, causing direct economic losses of tens of billions of yuan. Cause great loss to our pig industry. As ELISA is cheap and easy to operate, OIE regards it as the preferred serological method for ASF detection. P54 protein has good antigenicity and is an ideal antigen for detection. Objective: To identify a conservative site in the African swine fever virus (ASFV) p54 protein and perform a Cloth-enzyme-linked immunosorbent assay (ELISA) for detecting the ASFV antibody in order to reduce risks posed by using the live virus in diagnostic assays. Method: We used bioinformatics methods to predict the antigen epitope of the ASFV p54 protein in combination with the antigenic index and artificially synthesized the predicted antigen epitope peptides. Using ASFV-positive serum and specific monoclonal antibodies (mAbs), we performed indirect ELISA and blocking ELISA to verify the immunological properties of the predicted epitope polypeptide. Results: The results of our prediction revealed that the possible antigen epitope regions were A23-29, A36-45, A72-94, A114-120, A124-130, and A137-150. The indirect ELISA showed that the peptides A23-29, A36-45, A72-94, A114-120, and A137-150 have good antigenicity. Moreover, the A36-45 polypeptide can react specifically with the mAb secreted by hybridoma cells, and its binding site contains a minimum number of essential amino acids in the sequence 37DIQFINPY44. Conclusions: Our study confirmed a conservative antigenic site in the ASFV p54 protein and its amino acid sequence. A competitive ELISA method for detecting ASFV antibodies was established based on recombinant p54 and matching mAb. Moreover, testing the protein sequence alignment verified that the method can theoretically detect antibodies produced by pigs affected by nearly all ASFVs worldwide.

• The Korean Journal of Pharmacology
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• v.29 no.2
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• pp.263-274
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• 1993

Potassium $(K^+)$ channels are present in airway smooth muscle cells, and their activation results in hyperpolarization and relaxation. Because these effects may have therapeutic relevance to hypersensitivity and asthma, we examined the effect of a potassium channel activator, cromakalim (BRL 34915, CK) on the release of mediators from superfused tracheal and parenchymal strips after passive sensitization with $IgG_1$ antibody. Both tissues were superfused with CK $(2{\times}10^{-6}\;M)$ for 30 min and challenged with CK and antigen (Ox-HSA). Using monodispersed, partially purified, highly purified guinea pig lung mast cells, we also examined the effect of CK on mediator release from these cells after passive sensitization with $IgG_{1}$ antibody $({\alpha}-OA)$. Guinea pig lung mast cells were purified using enzyme digestion method, count current elutriation, and discontinuous Percoll density gradient. After CK pretreatment, passively sensitized mast cells were challenged with varying concentration of antigen (OA, immunological stimuli) or with varying concentration of calcium ionophore (CaI, non-immunological stimuli). Histamine (Hist) release was determined by spectrophotofluorometry, and leukotrienes (LT) by radioimmunoassy. CK pretreatment decreased Hist by 35% and LT release by 40% in the antigen-induced tracheal tissue after $IgG_1$ sensitization but did not decrease the contractile response. In the antigen-induced parenchymal tissue CK decreased Hist release by 25% but poorly decreased LT. Both immunologic and non-immunologic stimuli caused a dose-dependent release of Hist and LT from monodispersed, partially purified and highly purified lung mast cells. Verification of LT release was obtained by the use of 5-lipoxygenase inhibitor, A64077 (Zileuton). CK decreased Hist and LT release by 20% respectively in the OA-induced guinea pig lung mast cells after $IgG_1$ sensitization. The inhibitory effects of CK on the Hist and LT release in the Ox-HSA-induced airway smooth muscle tissues or in the OA-induced and CaI-induced mast cells after $IgG_1$ sensitization were completely blocked by TEA and GBC. These studies show that guinea pig lung mast cells seem to be an important contributor to LT release, and that CK (which has been known as an airway smooth muscle relaxant) can in part act to inhibit mediator release in the antigen-induced airway smooth muscle, and that CK may also act to inhibit mediator release in the OA-induced and CaI-induced highly purified mast cells. These results suggest that Hist and LT release evoked by mast cell activation might in part be associated with $K{^+}4 channel activity.

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

• The Korean Journal of Nuclear Medicine
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• v.26 no.2
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• pp.380-391
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• 1992

This study was designed to evaluate a direct method of $^{99m}Tc$ labeling using $\beta-mercaptoethanol$ as a reducing agent, and to investigate whether $^{99m}Tc$ labeled specific monoclonal antibody against carcinoembryonic antigen (CEA-92) can be used for the scintigraphic localization of human colon cancer xenograft. Purified CEA-92 IgG was fragmented into F $(ab')_2$ and then labeled with $^{99m}Tc$ by transchelation method using glucarate as a chelator. Labeling efficiency, immunological reactivity and in vitro stability of $^{99m}Tc$ CEA-92 F $(ab')_2$ were measured and then injected intravenously into nude mice bearing human colon cancer (SNU-C4). Scintigrams were obtained at 24 hour after injection. Then nude mice were sacrificed and the radioactivity was measured Labeling efficiency of injected $^{99m}Tc$ CEA-92 F $(ab')_2$, immunoreative fraction and in vitro stability at 24 hour of injected $^{99m}Tc$ CEA-92 F $(ab')_2$ was 45.2%, 32.8% and 57.4%, respectively. At 24 hour after injection, % ID/g in kidney (46.77) showed high uptake, but %ID/g in tumor (1.65) was significantly higher than spleen (0.69), muscle (0.16), intestine (0.45), stomach (0.75), heart (0.48) and blood (0.45). There was no significant difference between tumor and liver (1.81). Tumor contrast as quantitated by tumor to blood ratio of $^{99m}Tc$ CEA-92 F $(ab')_2$ was increased significantly (p<0.005) until 24 hours (3.70), and there was no statistical differece from tumor to blood ratio of I-131 CEA-92 F $(ab')_2$. The scintigram demonstrated localization of radioactivity over transplanted tumor, but significant background radioactivity was also noted over kidney and abdomen. It is concluded that CEA-92 F $(ab')_2$ can be labeled with $^{99m}Tc$ by a direct transchelation method using $\beta-mercaptoethanol$ as a reducing agent and $^{99m}Tc$ labeled CEA-92 F $(ab')_2$ can be used for the scintigraphic localization of human colon cancer xenograft in nude mice model.

• The Korean Journal of Nuclear Medicine
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• v.29 no.1
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• pp.98-104
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• 1995

Recently murine monoclonal antibodies have been studied actively for radioimmuno-scintigraphy and radioimmunotherapy, especially on patients with leukemia and lymphoma. In this research, we studied radiolabeling and immunologic characteristics of two in-house anti-leukemic monoclonal antibodies(anti-CALLA & anti-JL-1 antibodies) to make the basis for their clinical application. Each antibody was radiolabeled successfully with $^{99m}Tc$ by pretargeting transchelation method and with $^{125}I$ by lodogen method. We also studied cell binding assay, Scatchard analysis and modulation phenomenon. $^{125}I$ showed 90% labeling efficiency for each anti-body which was satisfactory, but $^{99m}Tc$ showed labeling efficiency below 70%, for which we need better labeling method. In cell binding assay, the immunoreactivity(IR) was low for $^{99m}Tc$-labeled antibodies. Scatchard analysis showed satisfactory data for both binding affinity. The affinity constant and antibody binding sites per cell are around $10^9M^{-1}$ and $10^4$, respectively. There was no modulation phenomenon in cases of $^{125}I$ or $^{99m}Tc$ labeled antibodies. We expect that two anti-leukemic monoclonal antibodies may be useful in diagnosis and therapy for leukemia and lymphoma patients.

• Journal of Microbiology and Biotechnology
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• v.16 no.9
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• pp.1347-1354
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• 2006

The aims of this study were to establish a simple and effective method for isolating male germline stem cells (GSCs), and to test the possibility of using these cells as a new approach for male infertility treatment. Testes obtained from neonatal and adult mice were manually decapsulated. GSCs were collected from seminiferous tubules by a two-step enzyme digestion method and plated on gelatin-coated dishes. Over 5-7 days of culture, GSCs obtained from neonates and adults gave rise to large multicellular colonies that were subsequently grown for 10 passages. During in vitro proliferation, oct-4 and two immunological markers (Integrin ${\beta}1,\;{\alpha}6$) for GSCs were highly expressed in the cell colonies. During another culture period of 6 weeks to differentiate to later stage germ cells, the expression of oct-4 mRNA decreased in GSCs and Sertoli cells encapsulated with calcium alginate, but the expression of c-kit and testis-specific histone protein 2B(TH2B) mRNA as well as the localization of c-kit protein was increased. Expression of transition protein (TP-l) and localization of peanut agglutinin were not seen until 3 weeks after culturing, and appeared by 6 weeks of culture. The putative spermatids derived from GSCs supported embryonic development up to the blastocyst stage with normal chromosomal ploidy after chemical activation. Thus, GSCs isolated from neonatal and adult mouse testes were able to be maintained and proliferated in our simple culture conditions. These GSCs have the potential to differentiate into haploid germ cells during another long-term culture.

• Korean Journal of Veterinary Research
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• v.5 no.1
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• pp.1-16
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• 1965

I. A Comparison of Sodium Sulfate Precipitation and Zone(Paper, Agar) Electrophoresis; Many kinds of techniques have been used for fractionating serum proteins. In the present study, using bovine serum, the fractions obtained with sodium sulfate were compared with those determined by zone electrophoresis. 1. Fibrinogen was precipitated with 4 to 10 percent of sodium sulfate. 2. ${\gamma}$-globulin required 10 to 16 percent of the salt for precipitation. 3. ${\beta}$-globulin began to precipitate at 12 percent sodium sulfate, and completed precipitation at approximately 26 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. 4. ${\alpha}$-globulin completed precipitation at 13 to 28 percent sodium sulfate in paper electrophoresis and at 22 percent in agar electrophoresis. 5. Albumin began to precipitate at 14 percent of the salt, and was free from the mixture of globulins approximately at 28 percent in paper electrophoresis, while at 22 percent in agar electrophoresis. The results of comparing fractions by the two methods were as follows: 1. Euglobulin (15%) was equal to the sum of the most ${\gamma}$-globulin and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. 2. Pseudoglobulin I (15-17.5%) corresponded to the most ${\alpha}$-, ${\beta}$-globulins and a small quantity of albumin. 3. Pseudoglobulin II(18-22%) was a mixture of the ${\alpha}$-, ${\beta}$-globulins and albumin fraction. 4. Albumin (above 22%) contained the most albumin fraction separated by zone electrophoresis and a small quantity of the ${\alpha}$-, and ${\beta}$-globulins. As mentioned above the fractions obtained with sodium sulfate were a mixture of the various proportion of the fractions determined by zone electrophoresis. The solubility of serum fractions to sodium sulfate coincided with the mobility of those by zone electrophoresis. (By percent of sodium sulfate we mean gram of sodium sulfate contained in $100m{\ell}$ of solution). II. Immunological Studies on Serum Protein Fractions with Sodium Sulfate; In the previous report the fractions of bovine serum protein with sodium sulfate compared with those obtained by zone electrophoresis, and the findings were that the former contained various proportion components of the latter. In this study the author studied whether or not the fractions with sodium sulfate are simple component antigenically by immunoelectrophoresis and micro double diffusion test (Immuno-precipitation), using rabbit antiserum to bovine serum. In immunoelectrophoresis, normal bovine serum developed with rabbit antibovine serum showed about ten distinct precipitin arcs. The distribution of these arcs was as follows: 1 albumin, 2 ${\alpha}_1$-, 3 ${\alpha}_2$-, 2 ${\beta}_1$-, ${\beta}_2$-, and 1 ${\gamma}$-globulin (Fig. 7, 9). In micro double diffusion test, five to six precipitation bands could be seen between antigens and antibody, the order of the precipitation bands location is albumin, ${\alpha}$-, ${\beta}$-, and ${\gamma}$-globulin from the side of antiserum well (Fig.19). Frequently the ${\alpha}$-, and ${\beta}$-precipitation bands were separated into two or three precipitation bands, which indicated that these globuline are not a pure component antigenically as shown in immuno-electrophoresis. In both Immunological methods, the two ${\alpha}$-, ${\beta}$-precipitin arcs and bands appeared clear and strong, indicating that the two globulins reacted as strong antigens. The precipitate reaction of ${\gamma}$-globulin was shown at 12 to 16 percent sodium sulfate; ${\beta}$-globulin at 12 to 20 percent; ${\alpha}$-globulin at 12 to 22 percent (immuno-electrophoresis), at 12 to 26 percent (Diffusion); and albumin at above 22 percent. Antigenically euglobulin contained ${\gamma}$-, ${\beta}$-, and ${\alpha}$-globulins, Pseudoglobulin I and Pseudoglobulin II were composed of ${\alpha}$-, and ${\beta}$-globulins, and albumin was a mixture of ${\alpha}$-globulin and albumin determined by zone electrophoresis. The results indicated that the fractions of serum protein obtained by either method were constituents of various proteins antigenically except ${\gamma}$-globulin and albumin by Zone electrophoresis.

• Korean Journal of Microbiology
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• v.38 no.3
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• pp.205-212
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• 2002

This study was conducted to develop a method for direct determination of phagocytic activities in human circulatic systems and to measure the phagocytic activities in human leukocytes from the alcoholics, since phagocytic activity was considered to be very important in human immune mechanism at early stage for the health care of the alcoholics. The subjects for this study were 130 among which 95 males and 3 females were diagnosed as alcoholism and 32 was healthy blood donors. A thin layer of heat-killed Staphylococcus aureus Cowan I was placed on a plastic dish and reacted with whole blood to measure the phagocytic plaque formation by human leukocytes. In order to determine the health conditions of the subjects, some clinical laboratory tests, such as white blood cell counts, hemoglobin contents (Hgb), mean corpuscular volume of red blood cells(MCV), serum electrophoresis, B and T-lymphocytes, T-lymphocyte subtypes and phytohemagglutination test were also implemented. Compared to the non-alcoholism, new and old alcoholic inpatients showed statistically significant differences on levels of Hgb and MCV (p<0.05), but showed that T and B-lymphocyte numbers decreased and Helper T cell/Suppressor T cell ratio ($1.6{\pm}0.8$%) increased. Compared to non-alcoholism, phagocytic plaque activities of leukocytes from alcoholic patients decreased significantly and an unusual pattern in phagocytic plaque was observed, showing a strange body and chain shaped phagocytosis. Based upon these results, it is concluded that a phagocytic-plaques of Staphylococcus aureus Cowan I by leukocytes was very simple and useful method for the early immunological determination of phagocytic activities in alcoholic patients without requiring any special equipments.

• Proceedings of the SCSK Conference
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• 2003.09b
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• pp.26-31
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• 2003

Korean Ginseng is a medicinal herb which grows naturally in korea. an ancient country situated in north-eastern Asia. Its medical use was already well known to herb doctors in this region about five thousand years ago since the effectiveness of korean ginseng has been recognized through practical use for a long time. Korean Ginseng has always been regarded as a devine cure. The name "Ginseng" can be found in various medicinal books. many of which were written as early as B.C. 100. In the records of many chinese medical books. dating from the inception of publishing, it was noted that Korean Ginseng was of the highest level of quality. Korean Ginseng originally grew in the mountains of korea. However, this wild Korean Ginseng(js called SANSAM) could not meet the ever-increasing demands. and from the 16th century. it has been cultivated on farms for mass processing and supplying in korea(js called INSAM). It was already recognized in korea a long time ago(B.C. 57 - A.D. 668) that Korean Ginseng possessed the qualities of panacea, tonic and rejuvenator, and had other medicinal properties as well. The effectiveness of Korean Ginseng is widely recognized among south-eastern Asians as well as Chinese. As its effect has been proved scientifically. Korean Ginseng is now becoming the ginseng for all human beings in the world. Korean ginseng is differently called according to processing method. Dried thing is Insam(white ginseng), boiled or steamed is Hongsam(red ginseng). 장뇌삼(long headed ginseng) is artificially grown in the mountain no in field for a long time. So the body is thin and some long. but ingredients are concentrated. Korean wild ginseng(SANSAM) is rare in these days but we developed cosmetic ingredient. The scientific name of Korean Ginseng is Panax Ginseng. It has acknowledge as a natural mysterious cure among the notheastern peoples. because of its broad medicinal application. The origin of the word" Panax" derived from panacea. a Greek word meaning cure-all. According to the classification method of herb medicines in the Chinese medicinal book. "God-Farmer Materia Medica(A.D. 483-496) korean Ginseng was described as the superlative drug: panacea. tonic and rejuvenator. We studied skin immunological effect. collagen synthesis. cell growth and whitening effect of SANSAM extract. IN cosmetics.. SANSAM extract had skin fibroblast cell growth effect. recover damaged skin in the sun and protect fine wrinkle. Also. In hair product.. inhibits hairless, white hair.its hairless, white hair.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.5
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• pp.706-716
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• 2014

A new strategy of co-inoculating Bacillus subtilis MA139 with Streptococcus thermophilus and Saccharomyces cerevisiae was used to produce fermented soybean meal (FSBM). Three experiments were conducted to determine the concentration of digestible energy (DE) and metabolizable energy (ME) (Exp. 1), apparent ileal digestibility (AID) and standardized ileal digestibility (SID) of amino acids (AA) (Exp. 2), and feeding value (Exp. 3) of FSBM produced by this new strategy (NFSB) compared with soybean meal (SBM) and conventionally available FSBM (Suprotein). In Exp. 1, twenty-four barrows (initial body weight [BW] of $32.2{\pm}1.7kg$) were randomly allotted to 1 of 4 diets with 6 replicates per diet. A corn basal diet and 3 diets based on a mixture of corn and 1 of 3 soybean products listed above were formulated and the DE and ME contents were determined by the difference method. The results showed that there were no differences in DE and ME between SBM and either FSBM product (p>0.05). In Exp. 2, eight barrows (initial BW of $26.8{\pm}1.5kg$) were fitted with ileal T-cannulaes and used in a replicated $4{\times}4$ Latin square design. Three corn-starch-based diets were formulated using each of the 3 soybean products as the sole source of AA. A nitrogen-free diet was also formulated to measure endogenous losses of AA. The results showed that the SID of all AA except arginine and histidine was similar for NFSB and SBM (p>0.05), but Suprotein had greater (p<0.05) SID of most AA except lysine, aspartate, glycine and proline than NFSB. In Exp. 3, a total of 144 piglets (initial BW of $8.8{\pm}1.2$ kg) were blocked by weight and fed 1 of 4 diets including a control diet with 24% SBM as well as diets containing 6% and 12% NFSB or 12% Suprotein added at the expense of SBM. During d 15 to 28, replacing SBM with 6% NFSB significantly improved average daily gain (ADG) and average daily feed intake (ADFI) (p<0.05) for nursery piglets. During the overall experiment, ADG of piglets fed diets containing 6% NFSB was significantly greater (p<0.05) than that of piglets fed SBM. In conclusion, fermentation with the new strategy did not affect the energy content or the AID and the SID of AA in SBM. However, inclusion of 6% NFSB in diets fed to nursery piglets improved performance after weaning likely as a result of better nutritional status and reduced immunological challenge.