• Tuberculosis and Respiratory Diseases
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• v.48 no.3
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• pp.315-323
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• 2000

Background : An immunochromatographic assay (ICT Diagnostics) which facilitates the diagnosis of tuberculosis(TB) by detecting serum antibodies mainly directed against specific 38KDa of Mycobacterium tuberculosis has come into the market. The test consists of a cardboard folding device containing nitrocellulose strip and absorbent pads. The whole procedure is completed within 15 min and does not require any additional equipment. The test has been reported to be sensitive and specific in diagnosing active TB. Thus the test had been evaluated with sera from TB patients and TB-free subjects. Method : Sera from patients with active pulmonary tuberculosis(40 sputum positives for Mycobacterium tuberculosis, 79 sputum negatives, and 3 extrapulmonary tuberculosis) were obtained from the Double-Cross Chest Clinic of the Korean National Tuberculosis Association (KNTA) in Seoul. The control group consisted of TB-free 68 subjects(21 children under 7 years old and 47 healthy staff members of KNTA). Results : Nine out of 68(13.2%) TB-free controls had positive antibody response. Total 106 of 122(86.9%) radiologically active patients had positive antibodies while 16 (13.1%) showed negative reaction. Antibody was detected in 38 of 40(95.0%) sputum positive patients and 68 of 82(82.9%) sputum negative patients who were under the antituberculosis chemotherapy. The sensitivity and specificity were all 87% and the positive predictive value was 92.2% while the negative predictive value was 78.7%, when the prevalence of TB in the sample was 64.2%. Our results clearly show that the detection of antibodies which mainly react with the 38KDa antigen of M. tuberculosis is not suitable as the first-line method of diagnosis but considered only as an adjunctive test to standard techniques of tuberculosis diagnosis. when considering its high false positivity.

• Journal of Nutrition and Health
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• v.35 no.2
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• pp.192-200
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• 2002

Conjugated linoleic acid (CLA) is a collective term for positional and geometric isomers of octadecadienoic acid in which the double bonds are conjugated. CLA has anticancer activity in a variety of animal cancer models, and cis-9, trans-11 (c9t11) and trans-10, cis-12(t10c12) CLA are the most predominant isomers present in the synthetic preparations utilized in these animal studies. To compare the ability of c9t11, t10c12 and an isomeric mixture of CLA to inhibit TSU-Prl cell growth, cells were incubated in a serum-free medium with various concentrations of these fatty acids. The isomeric mixture inhibited cell growth in a dose-dependent manner (1-3 $\mu$M) with a 41 $\pm$ 1% inhibition observed at 3 $\mu$M concentration after 48 hours. T10c12 also inhibited cell proliferation in a dote-dependent manner, However, the efficacy and potency of this isomer was much greater than that of the isomeric mixture with a 49 $\pm$ 2% inhibition observed at 0.3 $\mu$M concentration after 48 hours. By contrast, c9t11 slightly increased cell proliferation. To determine whether the growth-inhibiting effect of CLA is related to the changes in production of insulin-like growth factors (IGF) and IGF-binding proteins (IGFBP) by these cells, serum-free conditioned media were collected. Immunoblot analysis of conditioned media using a monoclonal anti-IGF-II antibody showed that both the isomeric mixture and t10c12 inhibited secretion of both mature 7,500 Mr and higher Mr forms of pro IGF-II, whereas c9t11 had no effect. Ligand blot analysis with 125I-IGF-II revealed the presence of two types of IGFBPs : 24,000 Mr IGFBP-4 and 30,000 Mr IGFBP-6. The production of IGFBP-4 slightly decreased at the highest concentrations of the isomeric mixture and t10c12. These results indicate that CLA inhibits human prostate cancer cell growth, an effect largely due to the action of t10c12. The growth inhibition may result, at least in part, from decreased production of IGF-II and IGFBP-4 by these cells.

• Research in Plant Disease
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• v.16 no.3
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• pp.326-330
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• 2010

Three pome fruit viruses, Apple chlorotic leaf spot virus (ACLSV), Apple stem grooving virus (ASPV) and Apple stem pitting virus (ASGV) were detected in pear trees (Pyrus pyrifolia) using double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) in Ansung, Naju and Ulsan provinces of Korea. Infection rate of three viruses was 35.2% from 452 leaf samples of the three cultivars of pear trees. Also, each of three viruses was detected by reverse transcription polymerase chain reaction (RT-PCR) for a limited number of samples. Infection rate of three viruses was 86.3% from 233 leaf samples of the three pear cultivars. The virus infection rates by RT-PCR were much higher than ELISA. ASGV was prevailing on pear with 74.2%, whereas ASPV and ACLSV were found in 34.8% and 0.4% of tested samples, respectively. Symptoms caused by ASGV showed black spots of infected Niitaka cultivar leaves. The ACLSV, ASPV and ASGV isolates showed 83~94% sequence identity at a nucleotide level to other pome fruit virus isolates when analyzed by NCBI BLAST. Pome fruit viruses occurring in pear were ACLSV, ASPV and ASGV. This is the first report of pear trees infected ASPV in Korea.

• Journal of Nutrition and Health
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• v.34 no.2
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• pp.123-131
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• 2001

The purpose of this study was to investigate the effects of diet composition on serum leptin and lipids profiles in rats. At the baseline, seven 4-week-old Sprague-Dawley rats, male, were sacrificed and the remaining rats were divided into five groups and each group was fed one of the following five dietary regiments for 2 weeks and 6 weeks;the control diet AIN76(CAL, n=2l), high-carbonhydrate(rice)(HCR, n=2l), high-carbohydrate(flour)(HCF, n=2l), high-fat(corn oil)(HFO, n=2l), high-fat(beef tallow)(HFB, n=2l). Serum leptin was determined by a double antibody ELISA assay at the baseline(n=7), 6 week(n=35) and 10 week of age(n=70). At 6 weeks of age, the increase in the Food Efficiency Ratio(FER) was related to adipocyte hyperplasia in rats on HFB diets. The serum triglyceride, total cholesterol, and LDL-cholesterol increased significantly in HFB group, and decreased in HFO group compared to control group. The HFC group showed significant increase in serum triglyceride level compared with control group. After 2 weeks and 6 weeks, noticeably high increases in epididymal adipose tissue fat cell mass and numbers were observed with the HFB fed group. Serum leptin levels increased as body weight increased over the period of time(4weeks; 1.50$\pm$0.13ng/ml versus 10weeks; 2.08$\pm$-.13ng/ml). And this result shows that there are 193% higher in rats fed high fat-beef tallow diet than the control diet. Serum leptin levels of the HFB group(4.01$\pm$0.39mg/ml) were significantly higher than that of the HFO(2.06$\pm$0.5613ng/ml), CAL(2.08$\pm$0.1313ng/ml), HCR(2.41$\pm$0.2113ng/ml) and HCF(2.80$\pm$0.4713ng/ml) at p<0.05. The serum leptin concentration was positively correlated with the amount of epididymal fat pads(r=0.47 p<0.01), serum triglyceride(r=0.49, p<0.001), tatal cholesterol(r=0.48, p<0.001), LDL-cholesterol(r=0.58, p<0.001), atherogenic index(r=0.67, p<0.001), and inversely correlated with HDL-cholesterol(r=-0.65, p<0.001). In conclusion, the changes in composition of dietary fat and carbohydrate intake could affect changes in concentration of serum lipids and leptin. Especially, the high-fat diet with animal fat source could increase circulating leptin level. (Korean J Nutrition 34(2) : 123-131, 2001)

• Proceedings of the Korean Society of Embryo Transfer Conference
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• 2002.11a
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• pp.97-97
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• 2002

Human eryhropoietin (EPO) is acidic glycoprotein hormone that plays key role in hematopoiesis by facilitating differentiation of erythrocyte and formation of hemoglobin (Hb) and is used for the treatment of anemia. Human EPO is consist of 166 amino acids which is modified by three N-glycosylations (24, 38, 83) and single O-glycosylation (126). N-glycosylation is reported to be related to the cellular secretion and activity of EPO. In this study, we examined effects of mutagenesis in glycosylation site of recombinat hEPO for the cellular secretion during production from cultured CHO cell. We produced rhEpo which was cloned by PCR from human liver cDNA (TaKaRa) in cultured CHO cell. Using supernatant of the culture, ELISA assay and western analysis were performed. To estimate biological activity, 20IU of rhuEpo was subcutaneously injected into four ICR mice. After 8 days, HCT level was increased average 13 per cent, RBC was increased ca. 2${\times}$10$\^$6//${\mu}\ell$. In disease model Rat (anemia c-kit, WSRC-WS/WS), HCT was increased ca. 12%, RBC was increased ca. 1.6${\times}$10$\^$6//${\mu}\ell$. These results suggests that rhEpo we produced has biological activity. To remove glycosylation site by substituting 24, 38, 83, and 126th asparagine (or serine) with glutamic acid, overlapping -extension site-directed mutagenesis was performed. To add novel glycosylation sites, 69, 105th leucine was mutated to asparagine. Mutant EPO construct was transfected into CHO cell. Supernatant of the cell culture was analyzed using ELISA assay with monoclonal anti-EPO antibody (Medac, Germany). Since, several reports for mutagenesis of glycosylation sites showed case-by-case results, we examined both transient expression and stable expression. Addition of novel glycosylation sites resulted no secretion while deletion mutants had little effect except some double deletion mutants (24/83 and 38/83) and triple mutant. We suggest that not single but combination of glycosyl group affect secretion of EPO.

• Food Science of Animal Resources
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• v.37 no.1
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• pp.1-9
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• 2017

The rotavirus-induced diarrhea of human and animal neonates is a major public health concern worldwide. Until recently, no effective therapy is available to specifically inactivate the rotavirion particles within the gut. Passive immunotherapy by oral administration of chicken egg yolk antibody (IgY) has emerged of late as a fresh alternative strategy to control infectious diseases of the alimentary tract and has been applied in the treatment of diarrhea due to rotavirus infection. The purpose of this concise review is to evaluate evidence on the properties and performance of anti-rotavirus immunoglobulin Y (IgY) for prevention and treatment of rotavirus diarrhea in human and animal neonates. A survey of relevant anti-rotavirus IgY basic studies and clinical trials among neonatal animals (since 1994-2015) and humans (since 1982-2015) have been reviewed and briefly summarized. Our analysis of a number of rotavirus investigations involving animal and human clinical trials revealed that anti-rotavirus IgY significantly reduced the severity of clinical manifestation of diarrhea among IgY-treated subjects relative to a corresponding control or placebo group. The accumulated information as a whole depicts oral IgY to be a safe and efficacious option for treatment of rotavirus diarrhea in neonates. There is however a clear need for more randomized, placebo controlled and double-blind trials with bigger sample size to further solidify and confirm claims of efficacy and safety in controlling diarrhea caused by rotavirus infection especially among human infants with health issues such as low birth weights or compromised immunity in whom it is most needed.

• Korean Journal of Veterinary Research
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• v.46 no.3
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• pp.177-184
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• 2006

Experimental autoimmune encephalomyelitis (EAE) is a disease model of multiple sclerosis (MS) that is characterized by remittance and relapse of the disease and autoimmune and demyelinating lesions in the central nervous system (CNS). Autoimmune inflammation is maintained by secretion of a large number of protein. Previous studies have suggested that transcripts encoding osteopontin (OPN) are frequently detected in the mRNA population of MS plaques. To elucidate the functional role of OPN in initiation and development of EAE, we examined the expression and localization of OPN in the spinal cord during acute EAE. We demonstrated that OPN significantly increased at the early stage of EAE and slightly declined thereafter by western blot analysis. An immunohistochemical study revealed that OPN was constitutively expressed in some glial cells (microglia, astrocytes) of white matter and neurons in the CNS of control rats. OPN expression was shown to be increased in the same cells at the early and peak stage of EAE. To identity cells expressing OPN by double-immunofluorescence labeling, we labeled rat spinal cord sections for OPN with a monoclonal OPN antibody and with mAbs for astrocyte (GFAP), microglia/macrophage (OX42)-specific markers. The major cell types of OPN-expressing cells were activated astrocytes and microglia in the adjacent inflammatory lesions. Interestingly, OPN was mainly expressed in the end feet of astrocytes around vascular cell adhesion molecule-1 (VCAM-1) expressing endothelial cells of CNS blood vessel. These findings suggest that increased levels of OPN in activated glial cell may play an important role in the recruitment of inflammatory cells into the CNS parenchyma during EAE.

• 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.

• Journal of agricultural medicine and community health
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• v.46 no.3
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• pp.162-170
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• 2021

Objectives: This study was carried out to understand the seroprevalence and risk factors for severe fever with thrombocytopenia syndrome (SFTS) among the Korea National Park Service (KNPS) workers. Methods: We used the stored serum samples (763) and survey results collected from the previous investigation on scrub typhus and Lyme disease among the KNPS workers during 2016-2017. The serum samples were analyzed by double-antigen sandwich enzyme-linked immunosorbent assay, which was used to test the total antibody including IgG and IgM. Results: The SFTS seroprevalence among the KNPS worrkers was 1.4%. In multivariate logistic analysis, the national park exploration programs (odds ratio, 3.48; 95% confidence interval, 1.01-12.01) was significantly associated with the seroprevalence of SFTS. Conclusion: This study was the first serological study of SFTS among forestry workers in South Korea. Although the KNPS workers are at a high-risk group of SFTS, the prevention activities related to the working environment and habit was insufficient. Thus, systematic prevention education and training for the KNPS workers need to be strengthened.

• Korean Journal of Veterinary Research
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• v.36 no.3
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• pp.681-692
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• 1996

The present study was conducted to isolate Babesia gibsoni by culture method of the microaerophilous stationary phase(MASP) and analyse the antigenic properties of the parasite by SDS-PAGE and immunoblot. The results obtained were summarized as follows. The protozoan parasite Babesia gibsoni multiplied in canine erythrocytes in RPMI 1640 medium(pH7.0) containing 20 40% normal canine serum under the MASP condition of 5% CO2 and 95% air at $37^{\circ}C$ incubator. The levels of parasitaemia in the erythrocytes were shown more higher by exchanging the medium at 24 hours interval. Under the above condition of MASP, the percentage of parasitized erythrocytes(PPE) after incubation for 8 days increased about 14 times more than that in the initiation of the 1% infected canine erythrocyte culture. The parasites were purely isolated from the MASP culture of red blood cells collected from dogs infected with Babesia gibsoni naturally or artificially. Among the total of 36 canine(Pit-bullterier) blood samples the parasites were isolated from 17 cases(47.2%) in the MASP culture while the parasites were detected from 20 cases(56%) and 12 cases(33.3%), respectively, by indirect fluorescent antibody(IFA) test and direct light microscopy(DLM). On the other hand, Babesia gibsoni was isolated by MASP culture from 15 cases(75%) and 11 cases(92%) of positive cases of IFA and DLM, respectively. In the analysis of the erythrocytic merozoite(AEOM) antigen derived from infected dog approximately 11 antigenic bands in molecular weight of 130, 120, 97.4, 92, 80, 52, 50, 42, 36, 30 and 29 KDa were observed on SDS-PAGE. Antigenic bands in the endoerythrocytic merozoite(CEOM) antigen derived from infected erythrocyte (sediment) in MASP culture were much similar to those of AEOM bands. In the exoerythrocytic merozoite(CEEM) antigen derived from supernatant of the infected erythrocyte culture approximately 20 antigenic bands were observed and the molecular weight of the major bands among these were 140, 120, 114, 105, 96, 93, 92, 80, 60, 52, 50, 38, 36, 30, 24, 18.5 and 16 KDa. In the protein patterns of AEOM and CEOM antigen by immunoblot 15 bands were observed and these patterns were much similar between each other. The molecular weight of the major bands in the both antigens were 130, 120, 80, 60, 52, 50, 42, 30, 29, 18.5 and 16 KDa. Approximately 21 bands were observed in CEEM antigen and the molecular weight of the major bands were 140, 120, 96, 92, 85, 80, 76, 60, 52, 50, 37, 30, 24, 16 and 15 KDa. The specific antigenic bands in the artificially infected dogs were firstly observed at 3 weeks afrer inoculation of infected blood and these antigenic bands were maintained up to 18 months after inoculation. In the immunoblot of the sera of the splenectomized dogs the specific antigenic bands with the molecular weight of 93 KDa and 52 KDa, respectively, were observed weakly comparing to those of non-splenectomized dog. In immunoblot of the sera collected from the naturally infected dogs the antigenic bands were observed as same as those of artificially infected dogs while antigenic band of 29 KDa in some individual dog showed strongly. In comparison of immunoblot of the sera collected from dogs non-treated and treated with diminazene aceturate(7mg/kg, IM) after artificial infection no differences of antigenic bands were observed. In analysis of antigenic bands by digoxigenin glycan/protein double labeling, antigenic bands in the molecular weight of 106, 60 58, 36, 30 and 29 KDa were determined as glycoproteins.