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Enzymatic activity in the fecal droppings from the house dust mite has been postulated to contribute to the elicited allergic response. Screening dust mite extracts through 137,180 tetrapeptide fluorogenic substrates allowed for the characterization of proteolytic substrate specificity from the potential cysteine and serine proteases in the extract. The extract was further screened against a 4000 member peptide nucleic acid (PNA) encoded inhibitor library designed to target cysteine proteases using microarray detection. Affinity chromatography coupled with mass spectrometry identified Der p 1 as one of the proteases targeted by the PNA inhibitors in the dust mite lysate. A phenotypic readout of Der p 1 function in allergy progression was demonstrated by the inhibition of CD25 cleavage from T cells by dust mite extract that had been treated with the Der p 1 inhibitor identified from the PNA-encoded inhibitor library.  相似文献   
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Our current understanding of the role and regulation of protease activity in normal and pathogenic processes is limited by our ability to measure and deconvolute their enzymatic activity. To address this limitation, an approach was developed that utilizes rhodamine-based fluorogenic substrates encoded with PNA tags. The PNA tags address each of the substrates to a predefined location on an oligonucleotide microarray through hybridization, thus allowing the deconvolution of multiple signals from a solution. A library of 192 protease substrates was prepared by split and mix combinatorial synthesis. The methodology and validation of this approach for profiling proteolytic activity from single proteases and from those in crude cell lysates as well as clinical blood samples is described.  相似文献   
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Physics of the Solid State - The spectra of luminescence and magnetic circular polarization of luminescence of praseodymium–yttrium aluminum garnet Pr3+ : Y3Al5O12 (PrYAG) are studied in the...  相似文献   
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Micro-bioreactor array for controlling cellular microenvironments   总被引:3,自引:0,他引:3  
High throughput experiments can be used to spatially and temporally investigate the many factors that regulate cell differentiation. We have developed a micro-bioreactor array (MBA) that is fabricated using soft lithography and contains twelve independent micro-bioreactors perfused with culture medium. The MBA enables cultivation of cells that are either attached to substrates or encapsulated in hydrogels, at variable levels of hydrodynamic shear, and with automated image analysis of the expression of cell differentiation markers. The flow and mass transport in the MBA were characterized by computational fluid dynamic (CFD) modeling. The representative MBA configurations were validated using the C2C12 cell line, primary rat cardiac myocytes and human embryonic stem cells (hESCs) (lines H09 and H13). To illustrate the utility of the MBA for controlled studies of hESCs, we established correlations between the expression of smooth muscle actin and cell density for three different flow configurations.  相似文献   
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The Faraday effect (FE) of the rare-earth (RE) paramagnetic garnet crystals associated with magnetic-dipole (MD) transitions has been investigated theoretically over a wide temperature range 80 to 300 K. Over this temperature range, the ??gyromagnetic?? Verdet V gm (m) of the Eu3Ga5O12 (EuGG) is not a linear function of the magnetic susceptibility ?? measured along the [111] axis. We find that the nontrivial character of the temperature dependence of the MD Verdet constant V gm (m) is associated with the presence of a contribution to the Faraday rotation of the Van Vleck ??mixing?? by an external magnetic field of the Eu3+ electronic states. The optical absorption and magnetic circular dichroism (MCD) of Eu3+ ion in EuGG at 85 K for the MD transition 7 F 0 ?? 5 D 1 were measured along the [111] axis. We show that the appearance of the MCD feature observed for the absorption line near 526.6 nm for the 7 F 0 ?? 5 D 1 MD absorption band may be well-explained by the contribution of Van Vleck ??mixing?? between the Stark singlet states from the 7 F 0 and 7 F 1 multiplet manifolds mixed by the external magnetic field H.  相似文献   
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Doubly charged ion mass spectra have been obtained for 11 organophosphorus compounds. Methane has been used as a target gas to increase the probability of single electron transfer collisions in the first field-free region of an Hitachi RMU-7L mass spectrometer. In general, the spectra of organophosphorus compounds do not exhibit molecular ions but are dominated by fragment ions, many of which must be formed by rearrangement processes. A geometry-optimized self-consistent field molecular orbital method has been employed to compute energies and structural parameters for prominent ions. In addition, a diabatic curve crossing model has been used to examine the single electron transfer reactions responsible for intense ions in the doubly charged ion mass spectra. Appearance energies measured for ions prominent in the 2E spectra of organophosphorus compounds have ranged from 23 to 38 eV.  相似文献   
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Shear‐thinning hydrogels are useful for biomedical applications, from 3D bioprinting to injectable biomaterials. Although they have the appropriate properties for injection, it may be advantageous to decouple injectability from the controlled release of encapsulated therapeutics. Toward this, composites of hydrogels and encapsulated microgels are introduced with microgels that are fabricated via microfluidics. The microgel cross‐linker controls degradation and entrapped molecule release, and the concentration of microgels alters composite hydrogel rheological properties. For the treatment of myocardial infarction (MI), interleukin‐10 (IL‐10) is encapsulated in microgels and released from composites. In a rat model of MI, composites with IL‐10 reduce macrophage density after 1 week and improve scar thickness, ejection fraction, cardiac output, and the size of vascular structures after 4 weeks when compared to saline injection. Improvements are also observed with the composite without IL‐10 over saline, emphasizing the role of injectable hydrogels alone on tissue repair.  相似文献   
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