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801.
Sequence analysis of the Clostridium cellulolyticum endoglucanase-A-encoding gene, celCCA 总被引:9,自引:0,他引:9
The nucleotide sequence of a Clostridium cellulolyticum endo-beta-1,4- glucanase (EGCCA)-encoding gene (celCCA) and its flanking regions, was determined. An open reading frame (ORF) of 1425 bp was found, encoding a protein of 475 amino acids (aa). This ORF began with an ATG start codon and ended with a TAA ochre stop codon. The N-terminal region of the EGCCA protein resembled a typical signal sequence of a Gram-positive bacterial extracellular protein. A putative signal peptidase cleavage site was determined. EGCCA, without a signal peptide, was found to be composed of more than 35% hydrophobic aa and to have an Mr of 50715. Comparison of the encoded sequence with other known cellulase sequences showed the existence of various kinds of aa sequence homologies. First, a strong homology was found between the C-terminal region of EGCCA, containing a reiterated stretch of 24 aa, and the conserved reiterated region previously found to exist in four Clostridium thermocellum endoglucanases and one xylanase from the same organism. This region was suspected of playing a role in organizing the cellulosome complex. Second, an extensive homology was found between EGCCA and the N-terminal region of the large endoglucanase, EGE, from C. thermocellum, which suggests that they may have a common ancestral gene. Third, a region, which extended for 21 aa residues beginning at aa + 127, was found to be homologous with regions of cellulases belonging to Bacilli, Clostridia and Erwinia chrysanthemi. 相似文献
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Rieke Alten Hermann Gram Leo A Joosten Bvanden Wim Berg Joachim Sieper Siegfrid Wassenberg Gerd Burmester Piet van Riel Maria Diaz-Lorente Gerardus JM Bruin Thasia G Woodworth Christiane Rordorf Yannik Batard Andrew M Wright Thomas Jung 《Arthritis research & therapy》2008,10(3):1-9
Introduction
IL-1β is a proinflammatory cytokine driving joint inflammation as well as systemic signs of inflammation, such as fever and acute phase protein production.Methods
ACZ885, a fully human monoclonal antibody that neutralizes the bioactivity of human IL-1β, was generated to study the potent and long-lasting neutralization of IL-1β in mechanistic animal models as well as in a proof-of-concept study in patients with rheumatoid arthritis (RA).Results
The mouse IL-1 receptor cross-reacts with human IL-1β, and it was demonstrated that ACZ885 can completely suppress IL-1β-mediated joint inflammation and cartilage destruction in mice. This observation prompted us to study the safety, tolerability and pharmacodynamic activity of ACZ885 in RA patients in a small proof-of-concept study – the first to be conducted in humans. Patients with active RA despite treatment with stable doses of methotrexate were enrolled in this dose escalation study. The first 32 patients were split into four cohorts of eight patients each (six were randomly assigned to active treatment and two to placebo). ACZ885 doses were 0.3, 1, 3 and 10 mg/kg, administered intravenously on days 1 and 15. To explore efficacy within 6 weeks of treatment, an additional 21 patients were randomly assigned to the 10 mg/kg cohort, resulting in a total of 20 patients dosed with 10 mg/kg and 15 patients treated with placebo. There was clinical improvement (American College of Rheumatology 20% improvement criteria) at week 6 in the 10 mg/kg treatment group; however, this did not reach statistical significance (P = 0.085). A statistically significant reduction in disease activity score was observed after 4 weeks in the 10 mg/kg group. Onset of action was rapid, because most responders exhibited improvement in their symptoms within the first 3 weeks. C-reactive protein levels decreased in patients treated with ACZ885 within 1 week. ACZ885 was well tolerated. Three patients receiving ACZ885 developed infectious episodes that required treatment. No anti-ACZ885 antibodies were detected during the study.Conclusion
ACZ885 administration to methotrexate-refractory patients resulted in clinical improvement in a subset of patients. Additional studies to characterize efficacy in RA and to determine the optimal dose regimen appear warranted.Trial Registration
ClinicalTrials.gov identifier NCT00619905. 相似文献804.
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The Ti Plasmid of Agrobacterium tumefaciens Harbors an attM-Paralogous Gene, aiiB, Also Encoding N-Acyl Homoserine Lactonase Activity 总被引:1,自引:0,他引:1
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A. Carlier S. Uroz B. Smadja R. Fray X. Latour Y. Dessaux D. Faure 《Applied microbiology》2003,69(8):4989-4993
The Agrobacterium tumefaciens C58 genome contains three putative N-acyl homoserine lactone (acyl-HSL) hydrolases, which are closely related to the lactonase AiiA of Bacillus. When expressed in Escherichia coli, two of the putative acyl-HSL hydrolases, AttM and AiiB, conferred the ability to degrade acyl-HSLs on the host. In Erwinia strain 6276, the lactonases reduced the endogenous acyl-HSL level and the bacterial virulence in planta. 相似文献
807.
Polypyrrole-glucose oxidase biosensor. Effect of enzyme encapsulation in multilamellar vesicles on analytical properties 总被引:1,自引:0,他引:1
In this paper, we present the analytical properties of a new type of polypyrrole-based, enzymatic amperometric biosensor. It is produced by encapsulating the enzyme, glucose oxidase (GOx), into onion-type multilamellar vesicles (MLV). We compare its properties to those of a classical GOx-polypyrrole biosensor. When MLV are used to embed GOx in polypyrrole (PPy), GOx behaves as a Michaelis-Menten enzyme. Without MLV, a deviation to the Michaelis-Menten behaviour is observed for high glucose concentrations. Kinetics parameters of both types of biosensors are studied as a function of the surface charge synthesis: GOx encapsulation induces a 200-fold increase of the apparent maximal current (I(m)(app)) and a 10-fold increase of the apparent Michaelis constant (K(m)(app)). Sensitivity is improved by a factor of 5. GOx is also shown to be less sensitive to inhibiting ions (Cl(-)) when MLV are used. A residual amperometric response of 43% instead of 3% is measured. Finally, the long-term stability of biosensors is improved by the GOx encapsulation. All these results are partially explained by our previous study on the morphology of PPy films fabricated with GOx encapsulated into onion-type MLV (Olea et al., 2007). 相似文献
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Background
Reproductive systems of male moths contain circadian clocks, which time the release of sperm bundles from the testis to the upper vas deferens (UVD) and their subsequent transfer from the UVD to the seminal vesicles. Sperm bundles are released from the testis in the evening and are retained in the vas deferens lumen overnight before being transferred to the seminal vesicles. The biological significance of periodic sperm retention in the UVD lumen is not understood. In this study we asked whether there are circadian rhythms in the UVD that are correlated with sperm retention. 相似文献810.