• Tuberculosis and Respiratory Diseases
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• v.58 no.5
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• pp.459-464
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• 2005

연구 배경 : Adenosine deaminase (ADA)는 퓨린 (purine) 대사에 작용하는 효소로서 림 프구, 특히 T-림프구의 증식과 분화에 관여하며, 결핵성 흉수의 진단에 있어서 중요한 생화학적 표지자 중의 하나이다. 한편, 노인의 경우, T-림프구의 수와 기능의 감소에 의하여 면역 기능이 감소하는 것으로 알려져 있다. 이에 저자 등은 노인 결핵성 흉수 환자에서 흉수내의 ADA 수치가 젊은 환자에서보다 감소하는지를 조사하였다. 방 법 : 4년 동안 세브란스 병원에서 1) 흉수 결핵균 배양 양성 또는 2) 흉막 조직 검사상 결핵에 합당한 소견을 보여 결핵성 흉수로 진단받은 환자 80명을 대상으로 후향적으로 조사하였다. 65세를 기준으로 두 군으로 분류하였으며, 연령과 흉수 ADA 수치의 연관 관계를 독립 표본 t-검정 및 선형 회귀 분석을 이용하여 연구하였다. 결 과 : 80명의 환자 중 65세 이상은 21명 (26.3%)이었다. 흉수 내의 ADA 수치는 65세 이상 및 이하 군에서 각각 $71.2{\pm}27.6IU/L$, $68.5{\pm}5.8IU/L$ 이었다 (p=0.69). 선형 회귀 분석에서도 연령과 흉수 내의 ADA 수치는 상관 관계를 보이지 않았다 ($r^2=0.05$, p=0.59). 결 론 : 본 연구의 결과에 의하면, 결핵성 흉수의 진단에서 흉수 ADA 수치를 보조 지표로 사용하는 데 있어서, 노인환자에서도 젊은 환자와 동일한 임상적 유의성을 가지고 동일한 결정 수치 (cut-off value)를 적용할 수 있을 것으로 판단된다.

• BMB Reports
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• v.39 no.2
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• pp.167-170
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• 2006

Bovine adenosine deaminase undergoes a nondenaturational conformational change at $29^{\circ}C$ upon heating which is characterized by a large increase in heat capacity. We have determined the transition state thermodynamics of the conformational change using a novel application of differential scanning calorimetry (DSC) which employs very slow scan rates. DSC scans at the conventional, and arbitrary, scan rate of $1^{\circ}C/min$ show no evidence of the transition. Scan rates from 0.030 to $0.20^{\circ}C/min$ reveal the transition indicating it is under kinetic control. The transition temperature $T_t$ and the transition temperature interval ${\Delta}T$ increase with scan rate. A first order rate constant $k_1$ is calculated at each $T_t$ from $k_1\;=\;r_{scan}/{\Delta}T$, where $r_{scan}$ is the scan rate, and an Arrhenius plot is constructed. Standard transition state analysis reveals an activation free energy ${\Delta}G^{\neq}$ of 88.1 kJ/mole and suggests that the conformational change has an unfolding quality that appears to be on the direct path to the physiological-temperature conformer.

• Journal of Life Science
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• v.9 no.2
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• pp.62-66
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• 1999

To overexpress E. coli ADA in host strain E. coli M15, the expression plasmid pQEADD was constructed. To analyze the expression characteristics of ADA, a time course of the expression was first examined. The protein was not detected in no inducted in no induction samples. After the addition of IPTG, a band corresponding to the expected size of ADA was appeared. Its molecular weight was about 36,000 dalton. Maximum expression level was revealed when the cell cultured for 3∼4hrs after induction. This result indicated that the efficient expression of add can be achieved by induction at early logarithmic phase. The effect of different IPTG concentration on the degree of ADA expression was investigated. The expression levels of add were not largely affected by IPTG concentration. Location of overexpression ADA was checked out. A protein band corresponding to the ADA was seen in only crude extract B(insoluble protein). This result suggests that ADA is in E. coli M15. The molecular weight of ADA estimated by SDS-PAGE was approximately 36,000 Da.

• Bulletin of the Korean Chemical Society
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• v.30 no.11
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• pp.2523-2531
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• 2009

Effects of some diacid, diamine and dinitro aromatic compounds on the structure and activity of adenosine deaminase (ADA) were investigated by UV-Vis spectrophotometry in 50 mM phosphate buffer at pH = 7.5 and 27 ${^{\circ}C}$ and molecular docking studies. The results showed that all tested ligands are showing inhibition; five ligands are uncompetitive and other two ligands are mixed of competitive and noncompetetive inhibitors with majority of competitive behavior. For the later case analysis was done based on competitive inhibition. Diacids have larger size and higher inhibition constant ($K_I$) relative to others. A logical correlation between calculated free energy of binding and experimental values was obtained for un-competitive. Experimental and calculated data showed that competitive inhibitors are distributed near the active site of enzyme and form several cluster of ranks, whereas uncompetitive inhibitors bind to the enzyme-substrate complex and distributed far from the active site. Results of structure-activity relationship showed that, larger, more hydrophobe, less spherical and more aromatic ligands have higher inhibition constants.

• Journal of Microbiology and Biotechnology
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• v.9 no.4
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• pp.429-435
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• 1999

An adenosine 5'-monophosphate deaminase (AMP aminohydrolase, EC 3.5.4.6) was purified to homogeneity from the cell-free extract of Saccharomyces cerevisiae DKCTC7248. The molecular mass of subunit was estimated to be 80 kDa on SDS-PAGE, and that of the holoenzyme was shown to be 240 kDa by gel filtration. The isoelectric point of the enzyme (AMP deaminase D) was determined to be 6.2. The AMP deaminase D was specific towards AMP with an apparent $K_m$ value of 4.1 mM and a Hill coefficient, $n_H$, of 2.2. Both ATP and ADP were positive allosteric effectors of the AMP deaminase D: The apparent $K_{m}$ was decreased to 1.6 mM and 3.3 mM in the presence of 0.1 mM ATP and ADP, respectively, lowering $n_{H}$ to 1.0. Univalent cations like $K^+, Na^+ and Li^ +$ activated the enzyme but some divalent cations such as $Cu^{ 2+} and Cd^{2+}$ showed strong inhibitory effects. This enzyme displayed optimum activity at $30^{\circ}C$ and pH 7.0. In addition, it was stable up to $45^{\circ}C$ and over a wide pH range(pH 5.5-9.0). Amino acid sequences of its N-terminal region were analyzed to be ADYKMQMFADDA.

• Tuberculosis and Respiratory Diseases
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• v.44 no.3
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• pp.611-620
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• 1997

Objectives : The differentiation of tuberculous effusion from the other causes of exudative pleural effusion remained difficult even with aids of biochemical analyses and pleural biopsy. As the pathophysiology of tuberculous pleural effusion is an enhanced cell mediated immunity, Adenosine deaminase(ADA) and various eytokines including Inteferon-$\gamma$, tumor necrosis factor alpha(TNF-$\alpha$) are considered as useful diagnostic tools in differentiating exudative pleural effusion. The author would like to demonstrate the diagnostic usefulness of TNF-$\alpha$ in the differentiation of exudative pleural effusion, and compared the discriminating ability of TNF-$\alpha$ with ADA. Methods : Pleural fluids obtained from 80 patients (tuberculous : 39, malignant : 31, parapneumonic : 10) with exudate pleural effusions were processed for cell counts and biochemical analysis including ADA and TNF-$\alpha$. Results : Tuberculous pleural fluid showed higher levels of ADA and TNF-$\alpha$, $48.7{\pm}32.7U/L$ and $184.1{\pm}214.2pg/mL$ than that of non-tuberculous effusion $26.0{\pm}41.3U/L$ and $44.1{\pm}114.2pg/mL$, respectively (ADA, TNF-$\alpha$, p < 0.05, p < 0.01). Receiver operating characteristics(ROC) curves were generated for ADA and TNF-$\alpha$ and the best cut-off value for adenosine deaminase and TNF-$\alpha$were considered as 30U/L and 15pg/ml, respectively. Comparing the area under the ROC curves, there was no significant difference between ADA and TNF-$\alpha$. Conclusion : For the differential diagnosis of tuberculous pleural effusion from the other causes of exudative pleural effusions, TNF-$\alpha$ as well as ADA was considered as useful diagnostic method. However adding TNF-$\alpha$ to ADA has no further diagnotic benefit than ADA alone.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.7 no.3
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• pp.163-168
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• 1974

The present paper deals with the degradation of nucleotides in the muscle of sea mussel, Mytilus edulis, during drying. Three kinds of samples, raw, hot-air dried, and steamed-and-hot air dried were prepared and the contents of nucleotides were determined by ion exchange chromatography on columns of Dowex 1, X8. ATP and ADP were dominant in the raw muscle showed about $8{\mu}moles/g$, dry basis, respectively. The rate of degadation of ATP was very slow during drying compared with those of fish. The accumulation of ADP and AMP were observed during drying and the amount of total nucleotides (ATP+ADP+AMP) were not decreased remarkably by drying process. IMP was not detected in the all of the samples examined, however, the contents of inosine and hypoxanthine were increased during drying. In case of inosine contents, the hot-air dried sample marked an exceedingly high value equivalent to 8 times of the raw sample whereas steamed-and-hot air dried sample showed 2 times of raw samples.

• Journal of the Korean Society of Food Science and Nutrition
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• v.25 no.1
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• pp.46-52
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• 1996

The present study was undertaken to investigate the effect of the water extract of the puffer fish Fugu xanthopterus(FXH) on the alcohol induced hyperuricemia. The normal group and the FXH treated group showed no sigbificant changes in the levels of blood uric acid but, the blood uric acid significantly decreased in the FXh treated rats with 100mg/kg for two weeks compared to the ethanol treated group. There were no significant changes in the activities of uricase, adenosine deaminase, guanine deaminase, and purine uncleoside phosphorylase, among all the test group. But the activitis of liver xanthine oxidase were recovered to the normal level in ethanol +FXH treated group comparing to the ethanol treated group. Furthermore, ethanol+FXH treated rats showed the similar pattern in the levels of blood uric acid and urinary allantoin with normal group. These results indicate that the decreased blood uric acid by the FXH treatment of the alcohol induced hyperuricemia rats may result from decreased activity of hepatic xanthine oxidase.

• Tuberculosis and Respiratory Diseases
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• v.45 no.2
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• pp.388-396
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• 1998

Background: Etiologic diagnosis of pleural effusion is usually made by clinical characteristics, pleural fluid analysis and pleural biopsy. But, despite careful diagnostic study, the cause of pleural effusion cannot be found in about 20 percent of patients, especially in loculated pleural effusions. Tuberculous pleurisy is one of the most common cause of pleural effusion in Korea. But, pleural fluid culture for Mycobacterium tuberculosis are positive in only 20 to 30 percent of patients and typical pleural biopsy finding in less than 50 percent of patients with this disease. In recent studies, adenosine deaminse(ADA) and its isoenzymes were proposed to be a useful diagnostic tool for differential diagnosis of pleural effusion. We investigated the pattern of ADA and its iscenzyme activities in various cause of pleural effusions to evaluate the diagnostic value of measuring ADA and its isoenzymes. Method: We measured total ADA and its isoenzyme activities in pleural fluid and serum from 54 patients with pleural effusion(25 tuberculous pleural effusion, 10 parapneumonic effusion, 14 malignant pleural effusion, 5 transudative pleural effusion), including 5 loculated tuberculous pleural effusions and 6 loculated parapneumonic effusions. Total ADA activity was measured by the spectrophotometric method and ADA2 isoenzyme activity was measured with same method using EHNA, potent inhibitor of ADA1 isoenzyme activity. Result: Total ADA activity of tuberculous pleural effusion was higher than malignant pleural effusion(p<0.01), but no significant difference was found between tuberculous pleural effusion and parapneumonic effusion(tuberculous pleural effusion: $148.9{\pm}89.9IU/L$, parapneumonic effusion: $129.0{\pm}119.4IU/L$, malignant pleural effusion: $48.7 {\pm}39.7IU/L$). Percentage of ADA2 activity to total ADA activity(ADA2%) of pleural effusion of tuberculous pleurisy was higher than parapneumonic effusion(p<0.05). but no significant difference was found between tuberculous pleural effusion and malignant pleural effusion(tuberculous pleural effusion: $57.2{\pm}10.7%$, parapneumonic effusion: $35.9{\pm}17.8%$, malignant pleural effusion: $60.7{\pm}4.1%$). In loculated pleural effusion, ADA2% of tuberculous pleural effusion was higher than parapneumonic effusion(tuberculous pleural effusion: $53.3{\pm}3.9%$, parapneumonic effusion: $27.8{\pm}7.9%$). Conclusion: Measurement of ADA isoenzyme activity is useful for differentiating tuberculous pleural effusion from parapneumonic effusion, especially in loculated pleural effusion.

• Proceedings of the KSME Conference
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• 2008.11a
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• pp.1687-1689
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• 2008

In the detection of pathogenic microorganisms ATP-bioluminescence reaction is a fascinating method. ATP(adenosine triphosphate) is an energy source of all kinds of living organism and ATP-bioluminescence reaction uses this ATP. However, ATP exists not only in the cells but also outside the cells. Therefore ATP-bioluminescence reaction only with intracellular ATP is very important in pathogenic microorganism detection. Because of that reason we developed a microfluidic channel containing Dielectrophoretic zone which capture microorganisms and eliminating and washing extracellular ATP with ATP-degarading enzymes, adenosine phosphate deaminase and apyrase. Microorganisms are captured by pDEP force at the DEP electrode zone and only extracellular ATPs are washed and eliminated outside the zone.