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11.
Online biochemical detection of glutathione-S-transferase P1-specific inhibitors in complex mixtures
Kool J Eggink M van Rossum H van Liempd SM van Elswijk DA Irth H Commandeur JN Meerman JH Vermeulen NP 《Journal of biomolecular screening》2007,12(3):396-405
A high-resolution screening (HRS) technology is described, which couples 2 parallel enzyme affinity detection (EAD) systems for substrates and inhibitors of rat cytosolic glutathione-S-transferases (cGSTs) and purified human GST P1 to gradient reversed-phase high-performance liquid chromatography (HPLC). The cGSTs and GST P1 EAD systems were optimized and validated first in flow injection analysis (FIA) mode, and optimized values were subsequently used for HPLC mode. The IC(50) values of 8 ligands thus obtained online agreed well with the IC(50) values obtained with microplate reader-based assays. For ethacrynic acid, an IC(50) value of 1.8 +/- 0.4 microM was obtained with the cGSTs EAD system in FIA mode and 0.8 +/- 0.6 microM in HPLC mode. For ethacrynic acid with the GST P1 EAD system, IC(50) values of 6.0 +/- 2.9 and 3.6 +/- 2.8 microM were obtained in FIA and HPLC modes, respectively. An HRS GST EAD system, consisting of both the cGSTs and the GST P1 EAD system in HPLC mode in parallel, was able to separate complex mixtures of compounds and to determine online their individual affinity for cGSTs and GST P1. Finally, a small library of GST inhibitors, synthesized by reaction of several electrophiles with glutathione (GSH), was successfully screened with the newly developed parallel HRS GST EAD system. It is concluded that the present online gradient HPLC-based HRS screening technology offers new perspectives for sensitive and simultaneous screening of general cGSTs and specific GST P1 inhibitors in mixtures. 相似文献
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Bianca T. A. de Greef Janneke G. J. Hoeijmakers Emma E. Wolters Hubertus J. M. Smeets Arthur van den Wijngaard Ingemar S. J. Merkies Catharina G. Faber Monique M. Gerrits 《PloS one》2016,11(2)
Objective
Screening for Fabry disease in patients with small fiber neuropathy has been suggested, especially since Fabry disease is potentially treatable. However, the diagnostic yield of testing for Fabry disease in isolated small fiber neuropathy patients has never been systematically investigated. Our aim is to determine the presence of Fabry disease in patients with small fiber neuropathy.Methods
Patients referred to our institute, who met the criteria for isolated small fiber neuropathy were tested for Fabry disease by measurement of alpha-Galactosidase A activity in blood, lysosomal globotriaosylsphingosine in urine and analysis on possible GLA gene mutations.Results
725 patients diagnosed with small fiber neuropathy were screened for Fabry disease. No skin abnormalities were seen except for redness of the hands or feet in 30.9% of the patients. Alfa-Galactosidase A activity was tested in all 725 patients and showed diminished activity in eight patients. Lysosomal globotriaosylsphingosine was examined in 509 patients and was normal in all tested individuals. Screening of GLA for mutations was performed for 440 patients, including those with diminished α-Galactosidase A activity. Thirteen patients showed a GLA gene variant. One likely pathogenic variant was found in a female patient. The diagnosis Fabry disease could not be confirmed over time in this patient. Eventually none of the patients were diagnosed with Fabry disease.Conclusions
In patients with isolated small fiber neuropathy, and no other signs compatible with Fabry disease, the diagnostic yield of testing for Fabry disease is extremely low. Testing for Fabry disease should be considered only in cases with additional characteristics, such as childhood onset, cardiovascular disease, renal failure, or typical skin lesions. 相似文献14.
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Improved nitrogen removal by application of new nitrogen-cycle bacteria 总被引:14,自引:0,他引:14
Jetten Mike S.M. Schmid Markus Schmidt Ingo Wubben Mariska van Dongen Udo Abma Wiebe Sliekers Olav Revsbech Niels Peter Beaumont Hubertus J.E. Ottosen Lars Volcke Eveline Laanbroek H.J. Campos-Gomez Jose Luis Cole Jeff van Loosdrecht Mark Mulder Jan Willem Fuerst John Richardson David van de Pas Katinka Mendez-Pampin Ramon Third Katie Cirpus Irina van Spanning Rob Bollmann Annette Nielsen Lars Peter den Camp Huub Op Schultz Carl Gundersen Jens Vanrolleghem Peter Strous Marc Wagner Michael Kuenen J. Gijs 《Reviews in Environmental Science and Biotechnology》2002,1(1):51-63
In order to meet increasingly stringentEuropean discharge standards, new applicationsand control strategies for the sustainableremoval of ammonia from wastewater have to beimplemented. In this paper we discuss anitrogen removal system based on the processesof partial nitrification and anoxic ammoniaoxidation (anammox). The anammox process offersgreat opportunities to remove ammonia in fullyautotrophic systems with biomass retention. Noorganic carbon is needed in such nitrogenremoval system, since ammonia is used aselectron donor for nitrite reduction. Thenitrite can be produced from ammonia inoxygen-limited biofilm systems or in continuousprocesses without biomass retention. Forsuccessful implementation of the combinedprocesses, accurate biosensors for measuringammonia and nitrite concentrations, insight inthe complex microbial communities involved, andnew control strategies have to be developed andevaluated. 相似文献
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Bacteriocin Release Protein Triggers Dimerization of Outer Membrane Phospholipase A In Vivo 总被引:4,自引:0,他引:4 下载免费PDF全文
Bacteriocin release protein is known to activate outer membrane phospholipase A (OMPLA), which results in the release of colicin from Escherichia coli. In vivo chemical cross-linking experiments revealed that the activation coincides with dimerization of OMPLA. Permeabilization of the cell envelope and dimerization were characterized by a lag time of 2 h. 相似文献
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