• Title/Summary/Keyword: Adenosine Deaminase

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A Product Inhibition Study on Adenosine Deaminase by Spectroscopy and Calorimetry

  • Saboury, Ali Akbar;Divsalar, Adeleh;Jafari, Ghasem Ataie;Moosavi-Movahedi, Ali Akbar;Housaindokht, Mohammad Reza;Hakimelahi, Hosain
    • BMB Reports
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    • v.35 no.3
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    • pp.302-305
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    • 2002
  • Kinetic and thermodynamic studies have been made on the effect of the inosine product on the activity of adenosine deaminase in a 50 mM sodium phosphate buffer, pH 7.5, at $27^{\circ}C$ using UV spectrophotometry and isothermal titration calorimetry (ITC). A competitive inhibition was observed for inosine as a product of the enzymatic reaction. A graphical-fitting method was used for determination of the binding constant and enthalpy of inhibitor binding by using isothermal titration microcalorimetry data. The dissociation-binding constant is equal to $140\;{\mu}M$ by the microcalorimetry method, which agrees well with the value of $143\;{\mu}M$ for the inhibition constant that was obtained from the spectroscopy method.

A Comparison of Microtuberization Efficiency between Normal and Adenosine Deaminase Transgenic Potato Plantlets Cultured In Vitro (Adenosine Deaminase 형질전환식물체와 정상식물체간의 인공씨감자 형성비교)

  • 최경화
    • Korean Journal of Plant Resources
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    • v.11 no.3
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    • pp.252-256
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    • 1998
  • A Study was conducted to investigate comparison of in vitro tuberization between normal and transgenic potato plantlets harboring adenosine deaminase gene in potato cultivar of Desiree. In time course study of in vitro tuberization, the rate of tuberization in four lines were increased till 6 weeks. but maintained stil after 7 weeks. Microtuber initiation of transgenic lines, 43 and 39 were faster than other lines, but no difference was observed after 5 weeks compared with normal plantlets. In all transgenic lines, the majoirty of microtubers produced were small(less than 100 mg) and medium(100-200mg) size rather than large size(more than 200 mg). Among 4 lines , line 9 produced the highest number of microtubers per each culture vessel. The results of this experiment suggest that there is no significant difference in microtuber production efficiency between normal and transgenic potatoes.

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Kinetics of Intracellular Adenosine Deaminase to Substrate Analogs and Inhibitors in Aspergillus oryzae (Aspergillus oryzae의 세포내 효소인 Adenosine Deaminase의 기질 유사체와 억제물질에 대한 반응속도론적 분석)

  • Choi, Hye-Seon
    • Korean Journal of Microbiology
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    • v.32 no.1
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    • pp.84-90
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    • 1994
  • Kinetic parameters of various substrates and inhibitors were measured to elucidate the binding requirements of the active site of intracellular adenosine deaminase (ADA) in Aspergillus oryzae. 3'-Deoxyadenosine was the best substrate according to the value of relative kcat/$K_m$. Purine riboside was found to be the strongest inhibitor with the $K_i$ value of $3.7{\times}10^{-5}$M. Adenine acted neither as a substrate nor as an inhibitor, suggesting the presence of ribose at N-9 of adenosine was crucial to binding. ADA also catalyzed the dechlorination of 6-chloropurine riboside, generating inosine and chloride ions. Substrate specificity of 6-chloropurine riboside was 0.86% of adenosine. Purine riboside, a competitive inhibitor of ADA, inhibit the dechlorination with similar $K_i$ value, suggesting that the same binding site was involved in deamination and dechlorination. Among the sulfhydryl group reagents, mercurials, pchloromercuribenzoate (PCMB), mersalyl acid and $HgCl_2$ inactivated the enzyme. Mersalyl acid-inactivated ADA was reactivated by thiol reagents, but PCMB-inactivated enzyme was not. When ADA was treated with the mercurial reagents, the inhibition constants and inhibition patterns were determined. Each inhibition was competitive with substrate. The $K_i$ values of these mercurial reagents were lower in 10 mM phosphate buffer than in 100 mM phosphate buffer, showing phosphate dependency.

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Isolation of Actinomycetes Producing Extracellular Adenosin Deaminase (세포외 Adenosine Deaminase를 생산하는 방선균의 분리)

  • 전홍기;김태숙
    • Korean Journal of Microbiology
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    • v.28 no.1
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    • pp.83-89
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    • 1990
  • Two strains of actinomycetes producing extracellular adenosine deaminase, strain J-845S and strain J-326TK, were isolated from soil. Strain J-845S was gram-positive and non-acid-fast. This strain formed whitish, rod-shaped, smooth and non-motile spores on the aerial mycelium, and the spore chain was spiral. The hyphae of the mycelium branched abundantly. Cell wall chemotypes of the strain were of type I containing LL-diaminopimelic acids, and of phospholipid type II, and then strain J-845S was designated as Streptomyces sp.. Strain J-326TK was gram-positive and non-acid-fast. The hyphae of primary and aerial mycelium fragmented into irregular rod of coccus-like elements. The aerial mycelium either did not branch or sparsely branched. Cell wall composition was of type I and phospholipid type I. Thus, strain J-326TK was identified as Nocardioides sp.

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방선균으로부터 생산되는 adenosine deaminase의 저해제 연구

  • 김경자
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1993.04a
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    • pp.139-140
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    • 1993
  • 토양으로부터 Actinomycetes 분리: Actinomycetes는 항생물질이나 항암물질을 많이 생산하는 중요한 균주로 의학계나 농업분야에 널리 이용되고 있다. Adenosine deaminase 저해제를 생산하는 균주를 찾기 위해 토양으로부터 actinomycetes를 분리하였다 Suluble-starch-casein배지와 AS-1배지를 이용하였는데 이들은 탄소원으로 용해성 전분을 공통적으로 함유하고 있다. 나무밑이나 산, 언덕등의 건조한 토양이나 두엄이나, 두엄밑의 토양을 탄산칼슘을 처리하여 주었을 때 actinomycetes가 많이 생장하는 것으로 알려져 있으므로 이러한 흙들을 채취하여 actinomycetes 분리에 이용하였다.

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Identification of Novel Mutations In Adenosine Deaminase Gene In Korean Leukemia Patients (한국인 백혈병 환자에서 아데노신 디아미나제 유전자의 새로운 변이의 확인)

  • Park, Ki-Ho
    • Journal of Life Science
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    • v.20 no.3
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    • pp.453-456
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    • 2010
  • Leukemia is the abnormal increase of hematopoietic progenitor cells in tissues, resulting in anemia, increased susceptibility to infection and impaired blood clotting. The adenosine deaminase (ADA) gene is an important druggable target for the treatment of leukemia patients. Genetic and molecular analyses were performed to determine the effects of ADA gene mutations in 20 leukemia patients in the Korean population. To analyze the relationship between genotype and phenotype, the ADA genomic DNAs - including 1,092 bp of 12 exons and partial intron sequences flanking each exon - were sequenced and compared. In this study, the known mutations in other diseases, more than 50 mutations already reported in patients with severe combined immunodeficiency disease (SCID) and autism, were not found, but two novel mutations in leukemia patients were discovered. They include one nonsense mutation (A>C at nt position 478, F101F) and one missense mutation (G>A at nt position 778, E260K). One missense mutation (G>A at nt position 22, D8Y) was also detected in 20 normal control patients (allelic frequency of 7.5%). Interestingly, subjects in the Korean population retained 2 bp insertion at the intron 6 (IVS6-52insGC), something that has never been shown in other populations. The genetic study to find out the correlation between the mutant alleles and leukemia patients revealed no association statistically (p>0.05). The mutation found in leukemia needs further study to determine its possibility as a molecular marker for the diagnosis of leukemia.

A Case of Mycoplasma Pneumoniae Pneumonia Accompanying High Adenosine Deaminase Activity in Pleural Effusion (흉막삼출액에서 높은 Adenosine Deaminase 활성도를 보인 마이코플라즈마 폐렴 1예)

  • Seo, Hyang-Eun;Kim, Yeon-Jae;Kim, Seong-Kyu;Kang, Hyun-Jae;Do, Yun-Kyung;Yoon, Hye-Jin;Chyun, Jae-Hyun;Lee, Byung-Ki;Kim, Won-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.52 no.1
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    • pp.70-75
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    • 2002
  • Mycoplasma pneumioniae has a unique genomic composition, cellular biology, and a fastidious nature as the smallest cell-free living organism that lacks a cell wall. Previous studies have suggested that a clinical manifestation of a M. pneumoniae infection is a consequence of a host immune response, particularly involving cellular immunity. Adenosine deaminase (ADA) is the main T-lymphocyte enzyme, and its activity is high in diseases where cellular immunity is stimulated. Therefore, its activity is useful for diagnosing a tuberculous pleural effusion. A pleural effusion is found in 5-20% of Mycoplasma pneumonia patients. However, there are few reports of high ADA activity in a mycoplasmal pleural effusion. Here we report a case of Mycoplasma pneumoniae infection established by a polymerase chain reaction and serologic tests, accompanying high ADA activity in a pleural effusion.

The Effect of Benzene on the Activity of Adenosine Deaminase in Tissues of Rats

  • Turhan, Ali;Dere, Egemen
    • BMB Reports
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    • v.40 no.3
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    • pp.295-301
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    • 2007
  • Swiss Albino (Rat rattus norvegicus) rats were intraperitoneally injected with a 100 mg $kg^{-1}$ dosage of benzene, a toxic and carcinogenic agent widely used for industrial purposes. Changes in the adenosine deaminase (ADA) activity in the liver, kidney and serum of rats were investigated at 0, 2, 4, 8, 16, 32 and 64 h following injection. Serum physiological was administered to each control group. Enzyme activities were measured spectrophotometrically. Our purpose was to further investigations of some diseases caused by benzene, and present evidence of variations in the activity of ADA enzyme effected by benzene. While benzene caused significant inhibitions in ADA activity in the liver at 16 and 32 h and at 0.05 probability level, no significant inhibition or activation occurred at other test periods (hours). ADA activity did not present any significant variation in the kidneys. It was observed that ADA activity displayed similar patterns in the control groups. Comparisons of ADA activities in the two groups showed a statistically significant decrease between $4^{th}$ and $64^{th}$ hours (p< 0.05), demonstrating a direct correlation between benzene and its effects on ADA enzymes.

Adenosine Deaminase - a Novel Diagnostic and Prognostic Biomarker for Oral Squamous Cell Carcinoma

  • Kelgandre, Deepak Chandrakant;Pathak, Jigna;Patel, Shilpa;Ingale, Pramod;Swain, Niharika
    • Asian Pacific Journal of Cancer Prevention
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    • v.17 no.4
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    • pp.1865-1868
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    • 2016
  • Background: The number of patients with oral cancer in India is increasing gradually (especially in younger people). Although the diagnostic modalities and therapeutic management of oral cancer are improving, the treatment outcome and prognosis of oral cancer remain poor. The absence of definite early warning symptoms for most head and neck cancers suggests that sensitive and specific biomarkers are likely to be important in screening for high-risk patients. Aims: To analyze serum adenosine deaminase (ADA) levels in oral squamous cell carcinoma (OSCC) cases who reported to our institute. Materials and Methods: A prospective study was performed on 100 histopathologically proven cases of OSCC (study group) and 100 normal healthy individuals (control group). Independent sample and one sample t-tests and one way ANOVA followed by Tuckey's POST HOC test were conducted for analysis. Results: Statistically significant increase in serum ADA levels was observed in OSCC cases compared to the control group. Also serum ADA level increased significantly with the histopathological grade. Conclusions: Serum ADA levels in OSCC may be a useful diagnostic and prognostic biomarkers in clinical practice and our findings suggest that a large-scale study is warranted to confirm clinical utility as a prognostic and diagnostic biomarker.

Diagnostic Accuracy of Lactate Dehydrogenase/Adenosine Deaminase Ratio in Differentiating Tuberculous and Parapneumonic Effusions: A Systematic Review

  • Larry Ellee Nyanti;Muhammad Aklil Abd Rahim;Nai-Chien Huan
    • Tuberculosis and Respiratory Diseases
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    • v.87 no.1
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    • pp.91-99
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    • 2024
  • Background: Tuberculous pleural effusion (TPE) and parapneumonic effusion (PPE) are often difficult to differentiate owing to the overlapping clinical features. Observational studies demonstrate that the ratio of lactate dehydrogenase to adenosine deaminase (LDH/ADA) is lower in TPE compared to PPE, but integrated analysis is warranted. Methods: We conducted a systematic review to evaluate the diagnostic accuracy of the LDH/ADA ratio in differentiating TPE and PPE. We explored the PubMed and Scopus databases for studies evaluating the LDH/ADA ratio in differentiating TPE and PPE. Results: From a yield of 110 studies, five were included for systematic review. The cutoff value for the LDH/ADA ratio in TPE ranged from <14.2 to <25. The studies demonstrated high heterogeneity, precluding meta-analysis. Quality Assessment of Diagnostic Accuracy Studies Tool 2 assessment revealed a high risk of bias in terms of patient selection and index test. Conclusion: LDH/ADA ratio is a potentially useful parameter to differentiate between TPE and PPE. Based on the limited data, we recommend an LDH/ADA ratio cutoff value of <15 in differentiating TPE and PPE. However, more rigorous studies are needed to further validate this recommendation.