• Tuberculosis and Respiratory Diseases
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• v.47 no.6
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• pp.757-767
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• 1999

Background: Diagnosis by smear and/or cultures of the Mycobacterium tuberculosis from body fluid or biopsy specimen is "Gold standard". However the sensitivity of the direct microscopy is relatively low and culture of mycobacteria is time consuming. Despite an explosion in the techniques of rapid identification of mycobacteria by molecular genetic means, it is laborious and expensive and then rapid, inexpensive serodiagnosis is interested in diagnosis of tuberculosis. But sensitivity and specificity of known serologic antigen is not full sufficient level and then new antigen develop and combination cocktails of new developed antigens by ELISA are needed. Method: To compare the efficacy of different mycobacterial specific antigen and to assess the applicability of the combination of several different antigens in the diagnosis of tuberculosis, five ELISA tests derived 14KDa, 16KDa, 19KDa, 23KDa, 38KDa were evaluated in 57 active pulmonary patient and 24 inactive post-therapy follow up patient and 48 normal control. Results: The optical densities of ELISA test with 14KDa, 16KDa, 19KDa, 23KDa, 38KDa were significantly higher in active tuberculosis cases than in normal control(P<0.001, P<0.001, P<0.027, P<0.001, P<0.001) and those with 16KDa, 38KDa were significant higher in active tuberculosis cases than in inactive post-therapy follow up cases(P<0.01. P<0.001) and those of 14KDa, 16KDa, 23KDa, 38KDa were significant higher in inactive post-therapy follow up cases than in normal control(P<0.008. P<0.01. P<0.006. P<0.001). The sensitivity of 14KDa, 16KDa, 19KDa, 23KDa, 38KDa in active pulmonary patient cases was 42.1%, 43.9%, 15.8%, 28.0%, 70.2%, respectively and the specificity of 14KDa, 16KDa, 19KDa, 23KDa, 38KDa in active pulmonary patient cases was 95.8%, 95.8%, 91.7%, 89.6%, 93.8%, respectively. The sensitivity and specificity of combination 38KDa with 16KDa was 87% and 93.7%. Conclusion: The sensitivity and specificity of new antigens for serodiagnosis of the tuberculosis still remains limited at around 70%, which makes its a poor diagnostic tool for disease confirmation. A combination of cocktail antigens provided by cut-off value adjustment for serodiagnosis of tuberculosis some improved diagnostic yield than single antigen serologic test.

• Proceedings of the PSK Conference
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• 2003.10b
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• pp.161.1-161.1
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• 2003

Two chitosanses produced by Aspergillus fumigatus KB-1 were purified by ion exchange and size exclusion chromatographies. Molecular weights of chitosanses were 111.23 KDa (chitosanase I) and 23.38 KDa (chitosanase II). The N-terminal amino acid sequence of chitosanase II was determined: YNLPNNLKQIYDKHKGKXSXVLAK(\ulcorner)GFTN. The optimum pH of the chitosanase I and II were 6.5 and 5.5 respectively. The optimum temperatures were $60^\times$C and $70^\times$. Two chitosanases were most stable at $10^\times$C. (omitted)

• Tuberculosis and Respiratory Diseases
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• v.38 no.3
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• pp.228-235
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• 1991

Pulmonary surfactant is a lipoprotein complex composed primarily of phospholipid and lungspecific apoproteins that reduces surface tension in the alveolus and maintains alveolar stability at low lung volume. Three families of lung-specific apoproteins have been described: SP-A, a glycoprotein with a reduced molecular weight of 28~36 KDa. SP-B a hydrophobic protein with a nonreduced molecular weight of 18 KDa, and SP-C a hydrophobic protein with a non-reduced molecular weight of 5~8 KDa. Surfactant proteins have important roles in regulating surfactant metabolism as well as in determining its physical properties. The synthesis of the active surfactant peptides appears to be modulated by system with considerable complexity, including numerous levels of regulation such as cell-specific, hormonal and developmental controls. Endotoxin appears to alter surfactant protein mRNAs differentially. It is hoped that the elucidation of the factors controlling the synthesis and metabolism of the surfactant proteins will aid in understanding the pathogenesis of hyaline membrane disease and offer new avenues for the therapy and diagnosis of ther pulmonary disorders as well.

• Journal of the Korean Chemical Society
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• v.38 no.8
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• pp.608-615
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• 1994

The isolation, purification and characterization of a carotenoprotein from the carapace of Pnaeus orientalis were investigated. The carotenoprotein was purple with broad λmax between 480, 409, 318 and 280 nm. Apparent structures were estimated by using X-ray diffractometry and scanning electron microscope, respectively. The molecular weight of the carotenoprotein complex had been determined by GPC and PAGE. The heavier complex, designated the $\alpha$-form (M.W = 170 KDa), was dissociated to a major subunit, $\beta$-form (M.W = 42 KDa). SDS-PAGE of $\alpha$-form showed apparently oligomeric pattern, and also $\beta$-form gave two polypeptides corresponding to 22 KDa and 19 KDa, respectively. The amino acid of the two proteins $({\alpha}-and\;{\beta})$-form, lipid and free fatty acid compositions were described. The prosthetic groups of the carotenoprotein were confirmed by TLC, IR, $^1H$-NMR, MS and various organic reactions as astaxanthin, astaxanthin monoester and astaxnathin diester.

• 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.

• Mycobiology
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• v.37 no.2
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• pp.121-127
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• 2009

Purification and characterization of intracellular cellulase produced by A. oryzae ITCC-4857.01 are reported. The enzyme was purified by ion-exchange chromatography using DEAE-cellulose followed by Gel filtration. The purification achieved was 41 fold from the crude extract with yield of 27%. The purified enzyme showed single band on poly acrylamide gel. The molecular weight as determined by SDS-PAGE and gel filtration was 38 KDa and 38.6 KDa respectively and contained only one subunit. The enzyme is glycoprotien as nature and contained 0.67% neutral sugar. The apparent Km value of the enzyme against cellulose was 0.83%. The enzyme showed the highest relative ativities on CMC followed by avicel, salicin and filter paper. The optimum pH of activity was 5.5 and very slight activity was observed at or above pH 7.5 as well as bellow pH 3.5. The optimum tempreture of the activity was $45^{\circ}C$ and the highest activity was exhibited in 35 to $45^{\circ}C$. The enzyme lost their activities almost completely (95${\sim}$100%) at $80^{\circ}C$ or above and as well as bellow $25^{\circ}C$.

• Journal of Plant Biotechnology
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• v.4 no.1
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• pp.39-44
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• 2002

The fast-migrating cationic peroxidase isozyme, named RC3, was purified from rice (Oryza sativa cv. Nak-Dong) callus. Purification of the enzyme was accomplished by ammonium sulfate fractionation, CM-cellulose ionexchange chromatography, and Sephacryl S-100 gel filtration. The molecular mass of the enzyme was about 34 KDa as determined by SDS-PACE and 38 KDa by Sephacryl-100 gel filtration. The pI value of the enzyme was 8.9. Antiserum against RC3 was raised in rabbits, and anti RC3 antiserum reacted with RC3 isozyme by Ouchterlony double immunodiffusion. The optimum pHs and Km values of the enzyme for various substrates were determined. Kinetic studies with various substrates showed that RC3 had very low Km value of 0.01 mM for ferulic acid and ascorbic acid. However, the enzyme did not use esculetin as a substrate.

• 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.

• Applied Biological Chemistry
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• v.43 no.1
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• pp.57-62
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• 2000

The proteoglycan, intracellular and extracellular, extracted from the liquid culture of Phellinus igniarius were purified and characterized. The mycelial productivity was proved to be better in shaking culture compared to standing culture. The productivity of intracellular proteoglycan of Phellinus igniarius appeared to be similar in two culturing methods. The standing culture of Phellinus igniarius produced 6 times as much extracellular proteoglycan compared to shaking culture. The proteoglycan were purified to a single peak by ion exchange chromatography(DEAE-cellulose) followed by gel filtration(Sepharose 2B). PIEPDG contained 79.0% total sugar and 7.2% protein. PIEPAG contained 56.7% total sugar and 40.8% protein. PIIPDG contained 64.8% total sugar and 17.4% protein. PIIPAG contained 56.9% total sugar and 41.5%n protein. The molecular weights of all the fractions were estimated to be above 100,000, from 134KDa of PIEPDG to 560 KDa of PIEPAG. The results of sugar analysis by HPLC showed that PIEPDG contains glucose only. The sugar part of PIIPDG and PIIPAG were consisted of glucose and inositol. The PIEPAG contained three kinds of monosaccharides, glucose, fructose and inositol.

• Journal of Korean Society on Water Environment
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• v.21 no.2
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• pp.125-131
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• 2005

Natural organic matter(NOM) is defined as the complex matrix of organic material and abundant in natural waters. It affects the performance of unit operations for water purification. Several kinds of analytical indicators such as DOC, specific ultraviolet absorbance(SUVA), apparent molecular weight (AMW), fractionation and high performance size exclusive chromatography(HPSEC) have been used to understand characteristics and variations of NOM. This study aims to evaluate the characteristics of NOM in the Geum River system comprising with stream flows and reservoirs. It was identified that SUVA denoting the portion of humic substance in water ranged within 1.60~3.36. Using resin adsorbents, dissolved organic carbon(DOC) was fractionated into three classes: hydrophobic bases(HOB), hydrophobic acids(HOA) and hydrophilic substances(HI). HI dominates in all samples, collectively accounting for more than 62% of the DOC. HOA was the second dominated fraction and it varied considerably but accounted for about 30% of the DOC. The distribution of high molecular weight(HMW) measured by HPSEC being used to determine the molecular weight distribution of aquatic humic substances was 40.1% and 38.7% in reservoir and stream flow, respectively. The distribution of low molecular weight(LMW) in stream flow was 13.2% higher than that in reservoir. And apparent molecular weight less than 1KDa, which include the molecular weight of hydrophilic organic matter, occupied with 69.2% and 68.2% in stream flow and reservoir, respectively. While the molecular weight of 1 to 100 KDa including humic substances ranged with 18.6% and 21.6% in stream flow and reservoir, respectively. Seasonal variation of refractory dissolved organic carbon was similar to that of SUVA.