• 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.

• Parasites, Hosts and Diseases
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• v.26 no.3
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• pp.155-162
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• 1988

Surface membrane proteins of virulent RH strain and tissue cyst-forming Fukaya strain of Toxoplasma gondii were analysed by SDS-polyacrylamide gel electrophoresis after LPO-catalyzed surface iodination and lectin blotting, then identified the zoite-specific antigens. Prior to the analyses, purification of RH tachyzoites from mouse peritoneal exudate and of Fukaya bradyzoites from mouse brain tissues were performed by centrifugation - on the discontinuous Percoll density-gradient. Ta- chysoites were obtained at the interface of 50U and 60% Percoll solution and brain cysts were harvested at the interfaces of 40-50% and 50-60%, then bradyzoites were obtained by treating the cysts with hypertonic solution. The LPO-catalyzed iodination detected 15 KDa and 14 KDa proteins o( brady- zoites and 30 KDa protein of tachysoites as major bands with several other minor bands. But Con A blotting revealed some bands of 200 K∼50 KDa glycoproteins of bradyzoites and 52 KDa band as major and minor bands of 33 K∼20 KDa of tachyzoites. Phytohemagglutinin did not detect any band in the two forms. EITB with anti- Fukaya antibody and anti-RH antibody revealed cross-reactivities between the two forms. Despite the cross-reactivity, anti-Fukaya antibody reacted with 15 KDa band of bradyzoites specifically and, anti-RH antibody with 52 KDa, 30 KDa, and 25 KDa bands of tachyzoites, respectively. It was identified that 15 KDa protein in bradyzoite, which was not a glycoprotein, was a major membrane protein with sufficient antigenicity, and in the case of tacky- zoite, 52 KDa surface glycoprotein (gp52) with specific antigenicity might be added to the major surface protein, p30.

• Archives of Pharmacal Research
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• v.17 no.5
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• pp.309-313
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• 1994

Phytase of rat intestine had a large amount of oilgosacchanrides ; The enzyme consisted of two different subunits with the molecular weights of 90 KDa and 70 KDa in its intack form, whereas the apparent molecular weights tumed to 72 KDa and 52 KDa, respectively, after deglycosylation. The treatment with glycopeptidase F reduced the molecular weights from 90 KDa and 70 KDa to 83 KDa and 52 KDa, respectively, While endoglycosidase H caused no change in their molecular weights. These results indicate that the 70KDa subunit contains only the N-linked oilgosaccharide chains, while the 90KDa subunit ocntains O-linked oilgosaccharides as well as N-linked ones. Enzyme-linked lectin assays suggeted that bisecting N-acetyl-D-glucosamine and galactose 1-4 N-acetyl-D-glucosamine structures were present and that fucose was included in these oilgosaccharide moieties. Sialic acid was not found in either subunit.

• Parasites, Hosts and Diseases
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• v.26 no.4
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• pp.245-254
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• 1988

We studied the serological reaction between various antigenic components from Cysticercus cellulosae and IgG antibodies in sera of cysticercosis, sparganosis, hydatidosis patients and normal humans by ELISA and EITB. In serological tests by ELISA, we recognized cross reaction of Cysticercus antigenic components with IgG antibodies in heterologous sera such as sparganosis and hydatidosis patients or normal humans. The crude antigenic components of Cysticercus showed lower ELISA sensitivity in homologous sera from cysticercosis patients than heterologous sera from hydatidosis patients. A total of 31 polypeptide bands with 260 KDa~22 KDa molecular weights were detected by SDS-PAGE, and 11 of them showed strong intensity. Total 22 components of them were recognized by IgG antibodies in cysticercosis patients sera. However, 12 of them were recognized also by normal human sera, 11 were by sparganosis sera, and-21 were by hydatidosis patients sera. The crude antigenic components of 104 KDa, 82 KDa, 72 KDa, 59 KDa and 34 KDa molecular weights were nonspecific ones, which cross-reacted with sera of either cysticerco, =is, sparganosis, hydatidosis patients or normal humans.

• KSBB Journal
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• v.14 no.5
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• pp.528-533
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• 1999

Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. CS5. The enzyme was solubilized and extracted with Trition-X and purified using the DEAE-Sephacel chromatography and Sephacryl S-200 chromatography. The enzyme was purified to 14-fold with a yield of 15%. The molecular weight of the purified enzyme was to be 332 KDa. SDS-PAGE of the enzyme showed three subunits with molecular weights of 79 KDa, 49KDa and 46K Da. It indicated that enzyme consisted of three subunits of the 79 KDa, two subunits of the 49 KDa and. 46 KDa, respectively. The apparent Km value for ethanol was 0.77 mM and the optimum pH and temperature was 4.0-5.0 and 35$^{\circ}C$, respectively.

• Korean Journal of Microbiology
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• v.37 no.4
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• pp.273-276
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• 2001

The strain which accumulate the silver in cell were isolated and characterized. And condition of accumulation of heavy metal was examined closely to investigate optimal condition of accumulation. Pseudomonas fluorescens and Bacillus cereus were Isolated from groundwater as the strain of silver accumulating bacteria. These strains did not grow in the medium at silver over the concentration 20 ppm. The largest accumulations of silver in the culture of Pseudomonas fluorescens and Bacillus cereus occurred within 24 hours. The amount of silver accumulation in Pseudomonas fluorescens and Bacillus cereus were 1.9 mg/g cell and 1.65 mg/g cell, repletively. In protein patterns of cell after the treatment of silver, three reducible proteins (126 KDa, 89 KDa, 25 KDa) in Bacillus cereus and one new protein (34 KDa) in Pseudomonas fluorescens were detected by SDS-PAGE.

• The Korean Journal of Zoology
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• v.35 no.4
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• pp.456-465
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• 1992

The 94 KDa glucose-resulsted Protein (SH 94), one of stress Proteins, is a Ca2+-binding protein in the endoplasmic reticulum (ER). In this study, the possible effect of Ca2+ on the native conformation of grp 94 was examined. When the purified grp 94 was analyzed by Sel filtration in the presence of either EGTA or CaCl2, it was eluted with apparent molecular weight (MW) of 100 KDa in both cases. When similarly analyzed with microtome or cell Ivsate, however, srp 94 was eluted with apparent IW of 200 KDa in the presence of E6TA, while with apparent MW of 100 KDa in the presence of CaCl2, indicating possible association of grp 94 with one or more other proteins in the absence of CaCl2. Consequently, immunoprecipitation with anti-grp 94 was carried out to determine which proteins specifically interact with grp 94. It is sho%un that srp 94 may interact, in a Ca2+_dependent manner. with other proteins including BiP (grp 78) which is also a stress protein in the ER.

• Journal of Food Hygiene and Safety
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• v.14 no.3
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• pp.270-276
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• 1999

In previous reports, the authors isolated the algicidal marine bacterium, Alteromonas sp. SR-14 and demonstrated its growth inhibition of diatom, Chaetoceros calcitrans (C. calcitrans). In this paper, we studied the effects of cell free culture filtrate of Alteromonas sp. SR-14 on the growth of C. calcitrans, and the characteristics of the algal growth inhibition substance. The culture filtrate of Alteromonas sp. SR-14 grown in peptone broth showed growth inhibition activity against C. calcitrans. The reasonable culture conditions of the bacterium for producing of algal growth inhibition substances were $15~20^{\circ}$ in temperature, 7.0-9.0 in pH and $23~30{\textperthousand}$ in salinity, respectively. The algal growth inhibition activity of culture filtrate was increased from stationary phase in growth curve of Alteromonas sp. SR-14. The molecular weights of algal growth inhibition substances produced by Alteromonas sp. SR-14 were ranged about from 3 KDa to 12 KDa. Among the substances, less than 10 KDa fraction were stable by heating at $100^{\circ}$ for 10 minutes, while more than 10 KDa fraction were heat labile. According to the experimental results, the algal growth inhibition substance produced by the bacterium was not a single compound.

• KSBB Journal
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• v.10 no.1
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• pp.30-37
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• 1995

Membrane-bound alcohol dehydrogenase(ADH) was purified to homogeneity from the acetic acid producing bacteria, Acetobacter sp. KM. The enzyme was solubilized and extracted with Triton X-100 and purified using the Mono-Q ion exchange chromatography and Superose 12 gel filtration chromatography. The enzyme was purified to 12-fold with a yield of 30%. The molecular weight of the purified enzyme was to be 335 KDa. SDS-PAGE of the enzyme showed two subunits with molecular weights of 79 KDa and 49 KDa. It indicated that the enzyme consisted of three subunits of the 79 KDa and two subunits of the 49 KDa. The purified .ADH preferentially oxidized straight chain aliphatic alcohol except methanol. Formaldehyde, acetaldehyde and glutaraldehyde were also oxidized. The apparent Km for ethanol was 1.04 mM and the optimum pH and temperature were 5.0∼6.0 and 32$^{\circ}C$, respectively. V2O5 and divalent cation such as ZnCl2 and NiCl2 inhibited enzymatic activity.

• The Korean Journal of Ecology
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• v.22 no.1
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• pp.39-43
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• 1999

Serratia marcescens, an enterobacterium of gram-negative bacteria, is characterized by resistance of the admium. Cadmium sensitive PM strain did not grow in the medium at cadmium concentration of 50 ppm. PA strain was induced to accommodate to cadmium by cultivating the mother strain (PC strain) in the medium with 50 ppm cadmium. As compared with PC and PM strains, PA strain revealed the excellent growth in cadmium media and accumulated four to five times higher cadmium concentration in cell than other strains. PA strain accumulated 23% of cadmium in cells when cultured in medium treated with 100 ppm cadmium and this cadmium was more accumulated in cytosol fractions than membrane fractions. Analysis by TEM indicated that cadmium was concentrated as a form of granule in cytosol. In protein patterns of cell after the treatment of cadmium, two inducible proteins (28 KDa and 64 KDa) and one reducible protein (45 KDa) were detected by SDS-PAGE. By Atomic Absorption Spectrophotometer, the amounts of cadmium attached to inducible proteins of 28 KDa and 64 KDa were 318.28 ㎍ and 325.37 ㎍ per gram of protein, respectively. It is assumed that these inducible proteins play an important role in the mechanism of cadmium accumulation in cells. A plasmid of 23Kbp was found in S. marcescens. The ability of resistance to cadmium in plasmid was confirmed by curing experiments.