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Deaminase

Distribution by Scientific Domains

Medical Sciences	45%
Chemistry	30%
Life Sciences	24%

Distribution within Medical Sciences

Immunology	20%
Gastroenterology & Hepatology	11%
Hepatology	6%
11 Other Subdomains	52%

Kinds of Deaminase

activation-induced cytidine deaminase

adenosine deaminase

cytidine deaminase

Terms modified by Deaminase

deaminase activity

Selected Abstracts

Activity of Adenosine Deaminase (ADA) and Adenylate Deaminase (AMPDA) Towards 6-Chloropurine Nucleosides Modified in the Ribose Moiety

EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 21 2004
Pierangela Ciuffreda
Abstract The enzymes adenosine deaminase (ADA) and adenylate deaminase (AMPDA) are able to catalyze the hydrolytic dechlorination of 6-chloropurine riboside and the corresponding 2,,3,- O -isopropylidene derivative, but show no activity towards the 3,4- O -isopropylidene-1-methylriboside of 6-chloropurine and adenine. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2004) [source]

Adenylate Deaminase (5,-Adenylic Acid Deaminase, AMPDA)-Catalyzed Deamination of 5,-Deoxy-5,-Substituted and 5,-Protected Adenosines: A Comparison with the Catalytic Activity of Adenosine Deaminase (ADA)

EUROPEAN JOURNAL OF ORGANIC CHEMISTRY, Issue 24 2003
Pierangela Ciuffreda
Abstract The enzyme adenylate deaminase (AMPDA) is able to catalyze the hydrolytic deamination of 5,-substituted and 5,-protected 5,-deoxyadenosines, whereas limited or no activity is shown by adenosine deaminase (ADA) towards the same substrates. (© Wiley-VCH Verlag GmbH & Co. KGaA, 69451 Weinheim, Germany, 2003) [source]

Modulation of Activation-Induced Cytidine Deaminase by Curcumin in Helicobacter pylori -Infected Gastric Epithelial Cells

HELICOBACTER, Issue 6 2009
Syed Faisal Haider Zaidi
Abstract Background:, Anomalous expression of activation-induced cytidine deaminase (AID) in Helicobacter pylori -infected gastric epithelial cells has been postulated as one of the key mechanisms in the development of gastric cancer. AID is overexpressed in the cells through nuclear factor (NF)-,B activation by H. pylori and hence, inhibition of NF-,B pathway can downregulate the expression of AID. Curcumin, a spice-derived polyphenol, is known for its anti-inflammatory activity via NF-,B inhibition. Therefore, it was hypothesized that curcumin might suppress AID overexpression via NF-,B inhibitory activity in H. pylori -infected gastric epithelial cells. Materials and Methods:, MKN-28 or MKN-45 cells and H. pylori strain 193C isolated from gastric cancer patient were used for co-culture experiments. Cells were pretreated with or without nonbactericidal concentrations of curcumin. Apoptosis was determined by DNA fragmentation assay. Enzyme-linked immunosorbent assay was performed to evaluate the anti-adhesion activity of curcumin. Real-time polymerase chain reaction was employed to evaluate the expression of AID mRNA. Immunoblot assay was performed for the analysis of AID, NF-,B, inhibitors of NF-,B (I,B), and I,B kinase (IKK) complex regulation with or without curcumin. Results:, The adhesion of H. pylori to gastric epithelial cells was not inhibited by curcumin pretreatment at nonbactericidal concentrations (,10 ,mol/L). Pretreatment with nonbactericidal concentration of curcumin downregulated the expression of AID induced by H. pylori. Similarly, NF-,B activation inhibitor (SN-50) and proteasome inhibitor (MG-132) also downregulated the mRNA expression of AID. Moreover, curcumin (,10 ,mol/L) has suppressed H. pylori -induced NF-,B activation via inhibition of IKK activation and I,B degradation. Conclusion:, Nonbactericidal concentrations of curcumin downregulated H. pylori -induced AID expression in gastric epithelial cells, probably via the inhibition of NF-,B pathway. Hence, curcumin can be considered as a potential chemopreventive candidate against H. pylori -related gastric carcinogenesis. [source]

Age-dependent variations of cell response to oxidative stress: Proteomic approach to protein expression and phosphorylation

ELECTROPHORESIS, Issue 14 2005
Yuri Miura Dr.
Abstract We investigated the protein profiles of variously aged rat astrocytes in response to oxidative stress. After H2O2 -exposure of cells at 100,µM for 30,min, the relative intensity of ten protein spots changed on two-dimensional (2-D) gels compared with control gels after silver staining. Matrix-assisted laser desorption/ionization-time of flight-mass spectrometry (MALDI-TOF-MS) analysis after in-gel digestion revealed that six of these spots corresponded to three kinds of proteins, each of which was composed of a protein and its modified form with a different isoelectric point (pI). These three proteins were identified as peroxiredoxins (PRDXs) II and III, and calpactin I light chain (p11). H2O2 -exposure increased the intensity of the spot with lower pI and simultaneously decreased that of the spot with higher pI for both PRDXs II and III. In addition, the expression of annexin VII, S -adenosyl- L -homocysteine hydrolase, elongation factor II fragment (EF-II), and adenosine deaminase was increased by H2O2 -exposure in astrocytes from variously aged rats. Using the Pro-Q® Diamond staining, heat shock protein 60,kDa (Hsp 60) and ,-tubulin were observed to be phosphorylated upon H2O2 -exposure. While phosphorylation of ,-tubulin was correlated positively with age, the changes in abundance of ten protein spots as described above were independent of age. These results suggest that aging does not suppress the responses aimed at limiting injury and promoting repair brought about by severe oxidative stress, and might affect cell dynamics including the formation of microtubules. [source]

Immunoregulatory effects of adenosine 5,-triphosphate on cytokine release from stimulated whole blood

EUROPEAN JOURNAL OF IMMUNOLOGY, Issue 3 2005
Els L. R. Swennen
Abstract In vitro studies suggest that extracellular nucleotides and nucleosides may be important regulators of inflammatory and immune responses. Most studies with adenosine 5,-triphosphate (ATP) have been performed in cell lines, which are remote from the human situation. The purpose of the present study was to determine the effects of ATP on TNF-,, IL-6 and IL-10 release in stimulated whole blood. Blood samples were drawn from healthy volunteers and incubated with ATP and lipopolysaccharide (LPS) + phytohemagglutinin (PHA) for 24,h. Contrary to expectations, ATP at 100,,M and 300,,M induced a reduction in TNF-, secretion by 32±8% (mean ± SEM) and 65±4%, respectively. Furthermore, these ATP concentrations induced an increase in IL-10 secretion by 48±5% and 62±7% in whole blood. The ATP analogue adenosine 5,-O-(3-thiotriphosphate) (ATP-,-S) and adenosine 5,-diphosphate (ADP) also inhibited TNF-, release, but only ADP showed a stimulatory effect on IL-10. Co-treatment with adenosine deaminase did not reverse the ATP effect on TNF-, and IL-10. These results show, for the first time, that ATP inhibits the inflammatory response in stimulated whole blood as indicated by inhibition of TNF-, and stimulation of IL-10 release and that this effect is predominantly mediated by ATP and not by adenosine. [source]

Activity of Adenosine Deaminase (ADA) and Adenylate Deaminase (AMPDA) Towards 6-Chloropurine Nucleosides Modified in the Ribose Moiety

Adenylate Deaminase (5,-Adenylic Acid Deaminase, AMPDA)-Catalyzed Deamination of 5,-Deoxy-5,-Substituted and 5,-Protected Adenosines: A Comparison with the Catalytic Activity of Adenosine Deaminase (ADA)

A novel coupled enzyme assay reveals an enzyme responsible for the deamination of a chemically unstable intermediate in the metabolic pathway of 4-amino-3-hydroxybenzoic acid in Bordetella sp. strain 10d

FEBS JOURNAL, Issue 15 2004
Chika Orii
2-Amino-5-carboxymuconic 6-semialdehyde is an unstable intermediate in the meta -cleavage pathway of 4-amino-3-hydroxybenzoic acid in Bordetella sp. strain 10d. In vitro, this compound is nonenzymatically converted to 2,5-pyridinedicarboxylic acid. Crude extracts of strain 10d grown on 4-amino-3-hydroxybenzoic acid converted 2-amino-5-carboxymuconic 6-semialdehyde formed from 4-amino-3-hydroxybenzoic acid by the first enzyme in the pathway, 4-amino-3-hydroxybenzoate 2,3-dioxygenase, to a yellow compound (,max = 375 nm). The enzyme in the crude extract carrying out the next step was purified to homogeneity. The yellow compound formed from 4-amino-3-hydroxybenzoic acid by this purified enzyme and purified 4-amino-3-hydroxybenzoate 2,3-dioxygenase in a coupled assay was identified as 2-hydroxymuconic 6-semialdehyde by GC-MS analysis. A mechanism for the formation of 2-hydroxymuconic 6-semialdehyde via enzymatic deamination and nonenzymatic decarboxylation is proposed based on results of spectrophotometric analyses. The purified enzyme, designated 2-amino-5-carboxymuconic 6-semialdehyde deaminase, is a new type of deaminase that differs from the 2-aminomuconate deaminases reported previously in that it primarily and specifically attacks 2-amino-5-carboxymuconic 6-semialdehyde. The deamination step in the proposed pathway differs from that in the pathways for 2-aminophenol and its derivatives. [source]

Identification and characterization of the genes for N -acetylglucosamine kinase and N -acetylglucosamine-phosphate deacetylase in the pathogenic fungus Candida albicans

FEBS JOURNAL, Issue 8 2001
Toshiko Yamada-Okabe
Like bacteria and many fungi, the pathogenic fungus Candida albicans can utilize GlcNAc as a carbon source for growth. A cluster of six genes was identified in the C. albicans genome. One of the genes in the cluster was CaNAG1, which is responsible for GlcN6P deaminase and is therefore essential for GlcNAc-dependent growth. The other five genes were designated CaNAG2, CaNAG3, CaNAG4, CaNAG5 and CaNAG6. The mRNA levels of CaNAG1, CaNAG2 and CaNAG5 were significantly induced by GlcNAc, whereas those of CaNAG3, CaNAG4 and CaNAG6 were not. Neither CaNAG2 nor CaNAG5 was essential for growth, but disruption of CaNAG2 or CaNAG5 greatly retarded the growth of cells using GlcNAc as the sole carbon source. Although no homolog of CaNAG2 or CaNAG5 was found in the Saccharomyces cerevisiae genome, CaNag2p displayed sequence similarities to Escherichia coli nagA, and CaNag5p is homologous to a wide variety of hexose kinases. When expressed as a fusion protein with glutathione S -transferase (GST), CaNag5p produced GlcNAc-P from GlcNAc in the presence of ATP, whereas GST alone did not. Furthermore, the recombinant GST,CaNag2p fusion protein converted GlcNAcP, which was produced by CaNag5p, into GlcNP. These results clearly demonstrate that CaNAG2 and CaNAG5 encode GlcNAcP deacetylase and GlcNAc kinase, respectively. CaNag5p recognized glucose and mannose as substrates, whereas the recently identified human GlcNAc kinase was specific to GlcNAc. Deletion of CaNAG2 or CaNAG5 markedly, and that of CaNAG1 moderately, attenuated the virulence of C. albicans in a mouse systemic infection model. Thus, it appears that GlcNAc metabolism of C. albicans is closely associated with its virulence. [source]

Cosmopolitan distribution of phlD -containing dicotyledonous crop-associated biocontrol pseudomonads of worldwide origin

FEMS MICROBIOLOGY ECOLOGY, Issue 2 2001
Chunxia Wang
Abstract In biocontrol fluorescent pseudomonads, phlD encodes a polyketide synthase required for the synthesis of the antifungal compound 2,4-diacetylphloroglucinol (Phl). Here, PCR-restriction fragment length polymorphism analysis was used to compare phlD alleles in 77 dicot-associated pseudomonads originating from various countries worldwide and 10 counterparts from a monocotyledonous host (wheat). The 16 restriction patterns obtained were mostly unrelated to geographic location or dicot host. Cluster analysis distinguished eight phlD clusters at a similarity level of 0.63. One cluster grouped 18 pseudomonads that produced also the antifungal polyketide pyoluteorin but could not assimilate D -galactose, D -galactonate lactone, D -sorbitol, L -arabinose, D -saccharate or D -xylose. These 18 pseudomonads, along with the eight pseudomonads from a second phlD cluster, were the only isolates that failed to deaminase 1-aminocyclopropane-1-carboxylate (ACC), a rare root growth promotion trait. Overall, assessment of phlD polymorphism, ACC deaminase activity and catabolic profiles pointed to a cosmopolitan distribution of Phl-producing biocontrol fluorescent pseudomonads of worldwide origin associated with dicotyledonous crop plants. [source]

Modulation of plant ethylene levels by the bacterial enzyme ACC deaminase

FEMS MICROBIOLOGY LETTERS, Issue 1 2005
Bernard R. Glick
Abstract Soil microorganisms that produce the enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase promote plant growth by sequestering and cleaving plant-produced ACC, and thereby lowering the level of ethylene in the plant. Decreased ethylene levels allows the plant to be more resistant to a wide variety of environmental stresses. Here, the biochemistry of ACC deaminase; the environmental distribution, regulation, evolution and expression of ACC deaminase genes; and information regarding the effect of this enzyme on different plants is documented and discussed. [source]

GANP suppresses DNA recombination, measured by direct-repeat ,-galactosidase gene construct, but does not suppress the type of recombination applying to immunoglobulin genes in mammalian cells

GENES TO CELLS, Issue 10 2007
Mikoto Yoshida
Immunoglobulin V-region somatic hypermutation and C-region class-switch recombination are initiated by activation-induced cytidine deaminase (AID) in B-cells. AID-induced DNA damage at the immunoglobulin S-region is known to be repaired by non-homologous end-joining, but repair mechanisms at the V-region remain to be elucidated. In Saccharomyces cerevisiae, DNA homologous recombination is regulated by the expression of Sac3, involved in actin assembly, cell cycle transition and mRNA metabolism. Here, we demonstrate that the Sac3-homologue GANP suppresses DNA recombination in a direct-repeat ,-galactosidase gene construct in mammalian cells. Homozygous ganp gene knockout is embryonic lethal in mice. Embryonic fibroblasts immortalized from hetero-deficient ganp+/, mice showed more DNA recombination than wild-type. In contrast, over-expression of GANP suppressed either spontaneous DNA recombination or that caused by the introduction of aid cDNA into NIH3T3 cells (susceptible to I-sceI restriction enzyme cleavage but not to RAG-mediated immunoglobulin gene recombination). GANP suppresses the DNA recombination not only on the extrachromosomal DNA construct but also on the integrated DNA. The Sac3-homology portion is necessary for the suppressive activity, but the truncated carboxyl terminal MCM3-binding/acetylating region adversely augmented DNA recombination, acting as a dominant negative form. Expression of full-length GANP is critical for suppression of DNA hyper-recombination in mammalian cells. [source]

Modulation of Activation-Induced Cytidine Deaminase by Curcumin in Helicobacter pylori -Infected Gastric Epithelial Cells

Induction of 150-kDa adenosine deaminase that acts on RNA (ADAR)-1 gene expression in normal T lymphocytes by anti-CD3-, and anti-CD28

IMMUNOLOGY, Issue 4 2007
Dama Laxminarayana
Summary We and other investigators have demonstrated up-regulation of the expression of the RNA-editing gene 150-kDa adenosine deaminase that acts on RNA (ADAR1) in systemic lupus erythematosus (SLE) T cells and B cells, peripheral blood mononuclear cells (PBMC), natural killer (NK) cells. The presence of a small proportion of activated T cells is the hallmark of SLE. Therefore, it was hypothesized that 150-kDa ADAR1 gene expression is induced by the physiological activation of T cells. To examine this hypothesis, normal T cells were activated by anti-CD3-, plus anti-CD28 for various time periods from 0 to 48 hr. The expression of 110-kDa and 150-kDa ADAR1, and interleukin (IL)-2 and ,-actin gene transcripts was analysed. An approximately fourfold increase in 150-kDa ADAR1 gene expression was observed in activated T cells. ADAR2 gene transcripts are substrates for ADAR1 and ADAR2 enzymes. Therefore, we assessed the role of the 150-kDa ADAR enzyme in editing of ADAR2 gene transcripts. In activated T cells, site-selective editing of the ,2 site was observed. Previous studies indicate that this site is predominantly edited by ADAR1. In addition to this, novel editing sites at base positions ,56, ,48, ,45, ,28, ,19, ,15, +46 and +69 were identified in activated T cells. On the basis of these results, it is proposed that 150-kDa ADAR1 gene expression is selectively induced in T cells by anti-CD3-, and anti-CD28 stimulation and that it may play a role in site-selective editing of gene transcripts and in altering the functions of several gene products of T cells during activation and proliferation. [source]

Therapeutic benefit of pentostatin in severe IL-10,/, Colitis

INFLAMMATORY BOWEL DISEASES, Issue 7 2008
Jeffrey B. Brown MD
Abstract Background: Pentostatin, an adenosine deaminase (ADA) inhibitor, is a purine antimetabolite used for the treatment of leukemias. ADA inhibition blunts expansion of proliferating lymphocytes and increases adenosine release, a potent anti-inflammatory molecule. Human inflammatory bowel disease (IBD) is driven by expansion of effector T cells (Teff) that overwhelm reulatory T cells (Treg) and propagate innate immune reponses. Here we study the therapeutic benefits of ADA inhibition to impair Teff cell expansion and reduce inflammatory cytokine release in IL-10-deficient (IL-10 -/- ) mice. Methods: Colitis was induced in IL-10 -/- mice by administering piroxicam for two weeks. Mice were treated with daily pentostatin or phosphate-buffered saline for 1 week and effects on tissue inflammation, lymphocyte numbers and cytokine production examined. Results: Pentostatin reduced inflammation by >50% and nearly normalized serum amyloid A levels. Lymphocyte expansions in the colon and mesenteric lymph node (MLN) (3.5-fold and >5-fold respectively) dropped by >50-90%. Pro-inflammatory factors in the colon and MLN (IL-1,, IFN-,, IL-6, CXCL10, TNF) dropped whereas FoxP3 and TGF-, were unchanged. Reductions in cytokine production from equivalent numbers of T cells from pentostatin-treated mice after in vitro (36h) or in vivo (3h) activation suggested anti-inflammatory effects of pentostatin independent of lymphodepletion contributed to its therapeutic benefit. Analysis of mucosal lymphocyte subsets suggested pentostatin reduced numbers of effector CD4+ CD69+ T cells, while sparing CD4+ CD62L+ T cells. Conclusions: Pentostatin dosages that avoid severe lymphocyte depletion effectively treat colitis by impairing Teff cell expansion and reducing pro-inflammatory cytokine production while preserving regulatory Treg populations and function. (Inflamm Bowel Dis 2008) [source]

Immune enhancement of nitroreductase-induced cytotoxicity: Studies using a bicistronic adenovirus vector

INTERNATIONAL JOURNAL OF CANCER, Issue 1 2003
Nicola K. Green
Abstract The nitroreductase (NR)/CB1954 enzyme prodrug system has given promising results in preclinical studies and is currently being assessed in phase I clinical trials. It is well established that there is an immune component to the bystander effect observed with other systems such as thymidine kinase and cytosine deaminase; however, such an effect has not previously been described using NR. We have preliminary data suggesting an immune bystander effect with NR to further examine these effects and their potential enhancement by cytokines, an adenoviral vector containing CMV-NR, an internal ribosome entry site (IRES) and the gene for murine GM-CSF (mGM-CSF) was constructed. The NR-GM-CSF virus was validated in 2 experimental models and demonstrated increased therapeutic efficacy in the MC26 murine colorectal tumour model. These data illustrate that the combination of suicide gene therapy using NR and CB1954 with immune stimulation via GM-CSF gives an improved response compared to either modality alone and suggests that the immune component of this response may be beneficial in combating unresectable, metastatic disease and preventing tumour recurrence. © 2002 Wiley-Liss, Inc. [source]

Diagnostic value of leptin in tuberculous pleural effusions

INTERNATIONAL JOURNAL OF CLINICAL PRACTICE, Issue 11 2006
G. CELIK
Summary It is suggested that leptin may be involved in inflammation. Although relation between leptin levels and active pulmonary tuberculosis has been studied, there is no information about relation between leptin levels and tuberculous pleural effusions (TPE). We evaluated the diagnostic value of pleural fluid and serum leptin levels in TPE and compared them with adenosine deaminase (ADA). Forty-five patients, 17 tuberculous effusion and 28 nontuberculous effusion, with exudative pleural effusions were included. Leptin and ADA levels were measured from serum and pleural fluid in all patients. There were no statistically significant differences between tuberculous and nontuberculous groups with respect to the serum ADA activity and pleural fluid/serum leptin ratio. On the contrary, pleural fluid leptin level, pleural fluid ADA activity, serum leptin level and pleural fluid/serum ADA activity ratio were statistically different between tuberculous and nontuberculous groups. When leptin levels were corrected for body mass index, serum leptin levels did not reach statistical significance. Cut-off points to predict tuberculosis were calculated as 9.85 ng/ml and 35.55 U/l for pleural fluid leptin level and pleural fluid ADA activity, respectively. Sensitivity, specificity and area under the curve ± standard error were 82.4%, 82.1%, 0.83 ± 0.07 for pleural fluid leptin levels and 100%, 100%, 1.00 ± 0.00 for pleural fluid ADA activity, respectively; the difference between these curves was significant (p = 0.01). Pleural fluid leptin levels were lower in tuberculous effusions than in other exudates. Pleural fluid leptin has a diagnostic value for TPE but not as good as that of ADA. [source]

Characterization of ACC deaminase gene in Pseudomonas entomophila strain PS-PJH isolated from the rhizosphere soil

JOURNAL OF BASIC MICROBIOLOGY, Issue 2 2010
Seralathan Kamala-Kannan
Abstract The enzyme 1-aminocyclopropane-1-carboxylate (ACC) deaminase cleaves the ethylene precursor ACC into ,-ketobutyrate and ammonia. The decreased level of ethylene allows the plant to be more resistant to a wide environmental stress including plant pathogens. In the present study, we characterized the ACC deaminase activity of a Pseudomonas entomophila strain PS-PJH isolated from the red pepper rhizosphere region of red pepper grown at Jinan, Korea. The isolate produced 23.8 ± 0.4 ,mol of ,-ketobutyrate/mg of protein/h during ACC deamination under in vitro conditions. Polymerase chain reaction for acdS gene showed that the isolated P. entomophila strain PS-PJH carry sequences similar to the known acdS genes. Results of the multiple sequence alignment revealed >99% identity (nucleotide and amino acid) with acdS gene of Pseudomonas putida strains AM15 and UW4. The isolated bacteria promoted 43.3 and 34.1% of growth in Raphanus sativus and Lactuca sativa plants, respectively. Based on the 16S,23S internal transcribed spacer region sequences, the isolate was identified as P. entomophila. To the best of our knowledge this is the first study to report the acdS gene in P. entomophila. (© 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source]

Deamination of adenosine by extracts of Penicillium politans NRC-510

JOURNAL OF BASIC MICROBIOLOGY, Issue 2 2005
Ali M. Elshafei Prof.
Cell-free extracts of nitrate-grown Penicillium politans NRC-510 could catalyze the hydrolytic deamination of adenosine to inosine maximally at pH 6.0 and 45 °C. However the same extracts could not catalyze the N-glycosidic bond cleavage of adenosine at pH 4.0, 6.0 and 8.0. Incubation of the extracts at 55 °C for 30 minutes caused about 31% loss in activity whereas incubation of the extracts at 60 °C for 15 minutes caused a complete loss of enzyme activity. Results indicated the absence of the involvement of sulfhydryl groups in the catalytic site of adenosine deaminase. The enzyme is inhibited by ethylene diamine tetraacetate indicating that adenosine deaminase is a metalloenzyme. MnCl2 and MgCl2 had a remarkable activating effect, whereas HgCl2, CaCl2 and ZnSO4 showed an inhibitory effect on enzyme activity. Dialyzing the extracts for 24 hours significantly increase deaminase activity by about 33%. The apparent Km value was calculated for adenosine and found to be 3.63 × 10,3M, which indicates high affinity of adenosine deaminase for its substrate adenosine. (© 2005 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim) [source]

Pleural fluid interferon-, and adenosine deaminase levels in tuberculosis pleural effusion: a cost-effectiveness analysis

JOURNAL OF CLINICAL LABORATORY ANALYSIS, Issue 2 2005
S.K. Sharma
Abstract Pleural fluid levels of interferon-, (IFN-,) and adenosine deaminase (ADA) have been found to be high in patients with tuberculosis (TB). The present study was carried out to compare the diagnostic utility of these two markers and to carry out a cost-effectiveness analysis of performing IFN-, estimation in comparison to ADA. A total of 52 patients with pleural effusion, 35 of which were found to have TB etiology, were prospectively included for estimation of ADA and IFN-, levels. The difference in the cost of performing the two diagnostic tests was compared with the cost of the treatment for a patient with TB. Pleural fluid IFN-, (median [range]: 2,100 [70,14,000] vs. 3 [0,160]; P<0.001) as well as ADA levels (mean [SD]: 93.1 [62.3] vs 15.4 [8.7]; P<0.001) were significantly higher in patients with TB effusion. Even though IFN-, estimation was more sensitive (97.1 vs. 91.4%), the extra cost of IFN-, estimation for detecting one patient with TB was found to be equivalent to the cost of a complete course of antituberculosis treatment for six patients. In developing countries, where TB is rampant and cost is a major concern, pleural fluid IFN-, estimation does not seem to be a cost-effective investigation method for differentiating TB from non-TB pleural effusion. J. Clin. Lab. Anal. 19:40,46, 2005. © 2005 Wiley-Liss, Inc. [source]

A theoretical study on the catalytic mechanism of Mus musculus adenosine deaminase

JOURNAL OF COMPUTATIONAL CHEMISTRY, Issue 12 2010
Xian-Hui Wu
Abstract The catalytic mechanism of Mus musculus adenosine deaminase (ADA) has been studied by quantum mechanics and two-layered ONIOM calculations. Our calculations show that the previously proposed mechanism, involving His238 as the general base to activate the Zn-bound water, has a high activation barrier of about 28 kcal/mol at the proposed rate-determining nucleophilic addition step, and the corresponding calculated kinetic isotope effects are significantly different from the recent experimental observations. We propose a revised mechanism based on calculations, in which Glu217 serves as the general base to abstract the proton of the Zn-bound water, and the protonated Glu217 then activates the substrate for the subsequent nucleophilic addition. The rate-determining step is the proton transfer from Zn-OH to 6-NH2 of the tetrahedral intermediate, in which His238 serves as a proton shuttle for the proton transfer. The calculated kinetic isotope effects agree well with the experimental data, and calculated activation energy is also consistent with the experimental reaction rate. © 2010 Wiley Periodicals, Inc. J Comput Chem, 2010 [source]

Acceleration of cartilage repair by genetically modified chondrocytes over expressing bone morphogenetic protein-7

JOURNAL OF ORTHOPAEDIC RESEARCH, Issue 4 2003
Chisa Hidaka
Background: Cartilage has a limited capacity to heal. Although chondrocyte transplantation is a useful therapeutic strategy, the repair process can be lengthy. Previously we have shown that over expression of bone morphogenetic protein-7 (BMP-7) in chondrocytes by adenovirus-mediated gene transfer leads to increased matrix synthesis and cartilage-like tissue formation in vitro. In this context we hypothesized that implantation of genetically modified chondrocytes expressing BMP-7 would accelerate the formation of hyaline-like repair tissue in an equine model of cartilage defect repair. Methods: Chondrocytes treated with adenovirus vector encoding BMP-7 (AdBMP-7) or as control, an adenovirus vector encoding an irrelevant gene (Escherichia coli cytosine deaminase, AdCD) were implanted into extensive (15 mm diameter) articular cartilage defects in the patellofemoral joints of 10 horses. Biopsies were performed to evaluate early healing at 4 weeks. At the terminal time point of 8 months, repairs were assessed for morphology, MRI appearance, compressive strength, biochemical composition and persistence of implanted cells. Results: Four weeks after surgery AdBMP-7-treated repairs showed an increased level of BMP-7 expression and accelerated healing, with markedly more hyaline-like morphology than control. Quantitative real-time polymerase chain reaction (PCR) analysis of the repair tissue 8 months after surgery showed that few implanted cells persisted. By this time, the controls had healed similarly to the AdBMP-7-treated defects, and no difference was detected in the morphologic, biochemical or biomechanical properties of the repair tissues from the two treatment groups. Conclusions: Implantation of genetically modified chondrocytes expressing BMP-7 accelerates the appearance of hyaline-like repair tissue in experimental cartilage defects. Clinical relevance: Rehabilitation after cell-based cartilage repair can be prolonged, leading to decreased patient productivity and quality of life. This study shows the feasibility of using genetically modified chondrocytes to accelerate cartilage healing. © 2003 Orthopaedic Research Society. Published by Elsevier Science Ltd. All rights reserved. [source]

Extractive components of boiled,dried scallop adductor muscle and effect on the taste of soup after mixing with chicken leg meat

JOURNAL OF THE SCIENCE OF FOOD AND AGRICULTURE, Issue 5 2005
Chie Yoneda
Abstract The extractive components of raw and boiled,dried scallop adductor muscle were examined. On a dry weight basis the levels of total free amino acids, total adenosine triphosphate (ATP) and related compounds, and betaines in the boiled,dried sample were lower than those in the raw muscle, which may have been due to the outflow of these compounds during boiling. Soup prepared from the boiled,dried scallop and chicken leg meat was assessed by sensory evaluation. This soup cooked with the scallop and chicken was stronger in sweetness, umami and body and rated higher in overall preference than the soup containing these ingredients after cooking separately. The inosine monophosphate (IMP) level in the former soup was 4.4 times higher than that in the latter. The adenosine monophosphate (AMP) deaminase (EC 3.5.4.6) activity in the crude extract from the chicken meat was 9.16 units l,1, whereas no activity could be detected in the crude extract from the boiled,dried scallop. It is concluded that AMP, which was mainly derived from the boiled,dried scallop, was converted to IMP by AMP deaminase from the chicken meat during the preparation of the soup, resulting in an improvement in the taste. Copyright © 2004 Society of Chemical Industry [source]

The large form of ADAR 1 is responsible for enhanced hepatitis delta virus RNA editing in interferon- , -stimulated host cells

JOURNAL OF VIRAL HEPATITIS, Issue 3 2006
D. Hartwig
Summary., Hepatitis delta virus (HDV) RNA editing controls the formation of hepatitis-delta-antigen-S and -L and therefore indirectly regulates HDV replication. Editing is thought to be catalysed by the adenosine deaminase acting on RNA1 (ADAR1) of which two different forms exist, interferon (IFN)- , -inducible ADAR1-L and constitutively expressed ADAR1-S. ADAR1-L is hypothesized to be a part of the innate cellular immune system, responsible for deaminating adenosines in viral dsRNAs. We examined the influence of both forms on HDV RNA editing in IFN- , -stimulated and unstimulated hepatoma cells. For gene silencing, an antisense oligodeoxyribonucleotide against a common sequence of both forms of ADAR1 and another one specific for ADAR1-L alone were used. IFN- , treatment of host cells led to approximately twofold increase of RNA editing compared with unstimulated controls. If ADAR1-L expression was inhibited, this substantial increase in editing could no longer be observed. In unstimulated cells, ADAR1-L suppression had only minor effects on editing. Inhibition of both forms of ADAR1 simultaneously led to a substantial decrease of edited RNA independently of IFN- , -stimulation. In conclusion, the two forms of ADAR1 are responsible almost alone for HDV editing. In unstimulated cells, ADAR1-S is the main editing activity. The increase of edited RNA under IFN- , -stimulation is because of induction of ADAR1-L, showing for the first time that this IFN-inducible protein is involved in the base modification of replicating HDV RNA. Thus, induction of ADAR1-L may at least partially cause the antiviral effect of IFN- , in natural immune response to HDV as well as in case of therapeutic administration of IFN. [source]

Proteasome- and SCF-dependent degradation of yeast adenine deaminase upon transition from proliferation to quiescence requires a new F-box protein named Saf1p

MOLECULAR MICROBIOLOGY, Issue 4 2006
Stéphanie Escusa
Summary In response to nutrient limitation, Saccharomyces cerevisiae cells enter into a non-proliferating state termed quiescence. This transition is associated with profound changes in gene expression patterns. The adenine deaminase encoding gene AAH1 is among the most precociously and tightly downregulated gene upon entry into quiescence. We show that AAH1 downregulation is not specifically due to glucose exhaustion but is a more general response to nutrient limitation. We also found that Aah1p level is tightly correlated to RAS activity indicating thus an important role for the protein kinase A pathway in this regulation process. We have isolated three deletion mutants, srb10, srb11 and saf1 (ybr280c) affecting AAH1 expression during post-diauxic growth and in early stationary phase. We show that the Srb10p cyclin-dependent kinase and its cyclin, Srb11p, regulate AAH1 expression at the transcriptional level. By contrast, Saf1p, a previously uncharacterized F-box protein, acts at a post-transcriptional level by promoting degradation of Aah1p. This post-transcriptional regulation is abolished by mutations affecting the proteasome or constant subunits of the SCF (Skp1,Cullin,F -box) complex. We propose that Saf1p targets Aah1p for proteasome-dependent degradation upon entry into quiescence. This work provides the first direct evidence for active degradation of proteins in quiescent yeast cells. [source]

Secretory antibodies against Giardia intestinalis in lactating Nicaraguan women

PARASITE IMMUNOLOGY, Issue 5 2005
A. Téllez
SUMMARY Secretory IgA (sIgA) antibodies are important in the host defence against the intestinal protozoan parasite Giardia intestinalis. However, few antigens have been identified. In this study 100 milk and saliva samples from lactating women, living in an endemic region (León, Nicaragua), were screened for the presence of antibodies against G. intestinalis. Most milk and saliva samples contained anti- Giardia antibodies (59% and 52%, respectively), with a mean sIgA content 50 times higher in milk than in saliva. The positive samples reacted with trophozoite membrane, flagella and cytoplasmic antigens. Western blot analysis showed that milk and saliva anti- Giardia sIgA recognized up to 16 different Giardia proteins in the molecular weight region 20,165 kDa. Two-dimensional Western blotting showed that the major immunoreactive proteins were the same as the immunoreactive proteins identified by serum from acute giardiasis patients in a non-endemic country. The major difference was a stronger reactivity against the variant surface proteins (VSPs) in the milk samples. Milk sIgAs also recognized recombinant Giardia proteins such as alpha-1 giardin, ornithine carbamoyl transferase, VSP-4EX, arginine deaminase and alpha-enolase. These antigens will be important targets in the development of new immunodiagnostic tools and vaccines. [source]

Six novel mutations of the ADAR1 gene in patients with dyschromatosis symmetrica hereditaria: Histological observation and comparison of genotypes and clinical phenotypes

THE JOURNAL OF DERMATOLOGY, Issue 7 2008
Taisuke KONDO
ABSTRACT Dyschromatosis symmetrica hereditaria (DSH), is a pigmentary genodermatosis of autosomal dominant inheritance. Since we clarified that the disease is caused by a mutation of the adenosine deaminase acting on the RNA 1 gene (ADAR1) in 2003, the molecular pathogenesis of a peculiar clinical feature of the disease has been expected to be clarified. We examined five familial cases and one sporadic case of Japanese families with DSH. The mutation analyses were done with single-strand conformation polymorphism/heteroduplex (SSCP/HD) analysis and direct sequencing of ADAR1. The DNA analysis of each patient revealed one missense mutation (p.F1091S), two nonsense mutations (p.C893X, p.S581X) and three frame-shift mutations (p.E498fsX517, p.F1091fsX1092, p.L855fsX856). Visual and electron microscopic findings showed abundant melanin pigment deposited all over the basal layer, and enlarged melanocytes with long dendrites located in the pigmented lesions with small or immature melanosomes scattered sparsely in the cytoplasm, but in the adjacent keratinocytes many small melanosomes were singly dispersed or aggregated. The hypopigmented areas showed little melanin deposition and reduced numbers of melanocytes in which much degenerative cytoplasmic vacuole formation could be observed by electron microscopy. Herein, we report six cases of DSH with six novel mutations. The variety of their clinical phenotypes even in the pedigree may suggest the presence of factors other than the ADAR1 gene influencing the extent of the clinical skin lesion. Microscopic findings suggest that the clinical appearance must have developed directly by melanocyte variations mainly induced by the ADAR1 gene mutations. [source]

Differential expression of activation-induced cytidine deaminase (AID) in nodular lymphocyte-predominant and classical Hodgkin lymphoma

THE JOURNAL OF PATHOLOGY, Issue 5 2005
Axel Greiner
Abstract Activation-induced cytidine deaminase (AID) is indispensable for class switch recombination and somatic hypermutation of immunoglobulin genes. Expression of AID has been detected in germinal centre centroblasts and in lymphomas derived from germinal centre cells. However, in situ studies of AID expression have until now been hampered by a lack of antibodies suitable for immunohistochemistry. To overcome this problem, an AID-specific monoclonal antibody suitable for immunohistochemical staining of formalin-fixed, paraffin wax-embedded tissue sections has been generated. This antibody was shown to detect AID expression in normal germinal centre B-cells as well as in non-Hodgkin lymphomas with a putative germinal centre origin. Using this antibody, a virtually exclusive cytoplasmic localization of AID in normal and neoplastic B-cells is shown. Employing a combination of immunohistochemistry and AID-specific in situ hybridization, it is demonstrated that AID is consistently expressed in the neoplastic cells of nodular lymphocyte-predominant Hodgkin lymphoma (HLnlp) but only infrequently in classical HL (cHL). This is in keeping with the notion that tumour cells of HLnlp represent transformed germinal centre B-cells showing evidence of somatic hypermutation. AID represents an additional marker useful in the differential diagnosis of HLnlp and cHL. Copyright © 2005 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd. [source]

ORIGINAL ARTICLE: Endogenous Adenosine Down-Modulates Mid-Trimester IntraAmniotic Tumor Necrosis Factor-, Production

AMERICAN JOURNAL OF REPRODUCTIVE IMMUNOLOGY, Issue 4 2009
Uma Perni
Problem, To determine whether adenosine in amniotic fluid down-regulates pro-inflammatory cytokine production. Method of study, Mid-trimester amniotic fluid from 21 women was incubated ex vivo in the presence or absence of human adenosine deaminase, the enzyme that irreversibly degrades adenosine. After 24 hr, supernatants were assayed by ELISA for tumor necrosis factor-, (TNF-,), interleukin (IL)-6, and IL-10. Clinical parameters were obtained after completion of laboratory testing. Results, Inclusion of adenosine deaminase resulted in a median increase in TNF-, production from 0.9 to 7.3 pg/mL (P = 0.0014). IL-6 production exhibited a non-significant median increase from <2.0 to 53.0 pg/mL (P = 0.0780). Median IL-10 production increased slightly from a median of <0.2 to 1.3 pg/mL. Adenosine deaminase-stimulated TNF-, production was proportional to parity and unrelated to gestational age, time of delivery, maternal age or indication for amniocentesis. Conclusion, Adenosine deaminase treatment increases TNF-, production by ex vivo -cultured amniotic fluid. Adenosine contributes to immune modulation in the amniotic cavity. [source]

Adenosine Deaminase Enzyme Therapy Prevents and Reverses the Heightened Cavernosal Relaxation in Priapism

THE JOURNAL OF SEXUAL MEDICINE, Issue 9 2010
Jiaming Wen MD
ABSTRACT Introduction., Priapism featured with painful prolonged penile erection is dangerous and commonly seen in sickle cell disease (SCD). The preventive approaches or effective treatment options for the disorder are limited because of poor understanding of its pathogenesis. Recent studies have revealed a novel role of excess adenosine in priapism caused by heightened cavernosal relaxation, and therefore present an intriguing mechanism-based therapeutic possibility. Aim., The aim of this study was to determine the therapeutic effects of adenosine deaminase (ADA) enzyme therapy to lower adenosine in priapism. Methods., Both ADA-deficient mice and SCD transgenic (Tg) mice display priapism caused by excessive adenosine. Thus, we used these two distinct lines of mouse models of priapism as our investigative tools. Specifically, we treated both of these mice with different dosages of polyethylene glycol,modified ADA (PEG,ADA) to reduce adenosine levels in vivo. At the end points of the experiments, we evaluated the therapeutic effects of PEG,ADA treatment by measuring adenosine levels and monitoring the cavernosal relaxation. Main Outcome Measures., Adenosine levels in penile tissues were measured by high-performance liquid chromatography, and cavernosal relaxation was quantified by electrical field stimulation (EFS)-induced corporal cavernosal strip (CCS) assays. Results., We found that lowering adenosine levels in penile tissues by PEG,ADA treatment from birth in ADA-deficient mice prevented the increased EFS-induced CCS relaxation associated with priapism. Intriguingly, in both ADA-deficient mice and SCD Tg mice with established priapism, we found that normalization of adenosine levels in penile tissues by PEG,ADA treatment relieved the heightened EFS-induced cavernosal relaxation in priapism. Conclusions., Our studies have identified that PEG,ADA is a novel, safe, and mechanism-based drug to prevent and correct excess adenosine-mediated increased cavernosal relaxation seen in two independent priapic animal models, and suggested its therapeutic possibility in men suffering from priapism. Wen J, Jiang X, Dai Y, Zhang Y, Tang Y, Sun H, Mi T, Kellems RE, Blackburn MR, and Xia Y. Adenosine deaminase enzyme therapy prevents and reverses the heightened cavernosal relaxation in priapism. J Sex Med 2010;7:3011,3022. [source]