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Molecular and genetic characterizations of five pathogenic and two non-pathogenic monoclonal antiphospholipid antibodies 总被引:3,自引:0,他引:3
Chukwuocha RU Zhu M Cho CS Visvanathan S Hwang KK Rahman A Chen PP 《Molecular immunology》2002,39(5-6):299-311
Antiphospholipid syndrome (APS) is an autoimmune disease that is characterized by thrombosis, recurrent fetal loss and thrombocytopenia. Antiphospholipid antibodies, detected by enzyme-linked immunoabsorbent assays (aCL) and/or in vitro blood clotting assays (LAC) are strongly associated with APS. Both the molecular structures used by pathogenic antiphospholipid antibodies and the genetic mechanisms leading to their production are unknown. We describe here the variable region genes of seven IgG antiphospholipid antibodies derived from two APS patients. Of these, five are pathogenic as defined in a mouse model of thrombosis and two are not. Analyses of the expressed variable region genes show no preferential V gene usage. However, similar to anti-DNA antibodies, pathogenic antiphospholipid antibodies contain an increased number of arginine residues in the third complimentarity-determining region (CDR3) of their H chains. The increased accumulation of arginine residues in the V(H) CDR3 may act to enhance antigen binding, promote disease and point to the importance of the H chain in the pathogenic potential of certain antiphospholipid antibodies. 相似文献
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Rong Liu Brian Hoffpauir Shannon D Chilewski Janice Gamberdella Uma Kavita Jia Duo Carol Gleason Yan Zhang Renuka Pillutla Binodh DeSilva Lora Hamuro 《The AAPS journal》2017,19(1):82-91
The Gyrolab? xP is a microfluidic platform for conducting ligand binding assays (LBAs) and is recognized for its utility in discovery bioanalysis. However, few reports have focused on the technology for regulated bioanalysis. This technology has the advantage of low reagent consumption, low sample volume, and automated ligand binding methods. To improve bioanalysis testing timelines and increase the speed at which biotherapeutics are delivered to patients, we evaluated the technology for its potential to deliver high-quality data at reduced testing timelines for regulated bioanalysis. Six LBA methods were validated to support bioanalysis for GLP toxicokinetic or clinical pharmacokinetic studies. Validation, sample analysis, and method transfer are described. In total, approximately 4000 samples have been tested for regulated bioanalysis to support 6 GLP toxicology studies and approximately 1000 samples to support 2 clinical studies. Gyrolab? xP had high run pass rates (≥83%) and high incurred sample reanalysis (ISR) pass rates (>94%). The maximum total error observed across all QC levels for a given assay was <30% for all six LBAs. High instrument response precision (CV ≤5%) was observed across compact discs (CDs), and methods were validated to use a single standard curve across multiple CDs within a Gyrolab? xP run. Reduced bioanalysis timelines were achieved compared to standard manual plate-based methods, and methods were successfully transferred across testing labs, paving the way for this platform for use in late-stage clinical development. 相似文献
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Aim: This paper describes the performance of 5th year medical students in multiple choice question (MCQ) examinations before and after a geriatric medicine teaching block. Methods: A 30‐question MCQ test was administered at the start and a 45‐question one at the end of the course. Results: There was a statistically significant improvement in the MCQ scores from a mean of 62% (SD 10.4) to 75.2% (SD 7.9) (P < 0.001). Total mean scores for the University of California, Los Angeles (UCLA) Geriatrics Knowledge test improved from 65% (SD 10.4) to 73%(SD 11.7) (P < 0.001). Total mean scores for the American Geriatric Society (AGS) Geriatrics Review Syllabus MCQs improved from 59.3% (SD 17.0) to 78.1% (SD 12.1) (P < 0.001). Post‐course, students scored equally well in the new questions, the validated UCLA test and the AGS questions. Conclusion: An undergraduate geriatric medicine clinical teaching block in senior clinical years can increase students' knowledge in geriatric medicine. 相似文献
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Mycobacterium immunogenum and Mycobacterium chelonae are closely related species associated with occupational hypersensitivity pneumonitis (HP) and nosocomial infections. There is a need to develop specific and readily adaptable methods for detection and speciation of these agents. Here we report development of a probe-based colorimetric-PCR assay involving heat shock protein (hsp) gene amplification (228bp) and its detection in an ELISA-like reaction. A quantitative format of this assay was developed and validated on metalworking fluids (MWF). The assay showed a minimum detection limit of 10 fg genomic DNA or 1 mycobacterial cell, albeit with variations depending on type and composition of the MWF matrix. When applied to the field MWF samples, the developed assay was found to be comparable to the real-time PCR assay, and allowed direct speciation of MWF mycobacteria without sequencing and/or restriction pattern analysis. In conclusion, the developed colorimetric PCR allows detection and quantification of MWF mycobacteria without culturing and is the first probe-based assay for unambiguous differentiation between the two phylogenetically closely related species, M. immunogenum and M. chelonae. Considering that the assay offers high throughput format involving relatively simpler instrument infrastructure, it has a potential for applications in routine assessment of MWF mycobacteria in diagnostic and industrial laboratories. 相似文献