Correlation between CYP2B6 and HNF3β expression in HPH.

<p>Total RNA was extracted from HPHs prepared from 35 human liver donors. Expression levels of CYP2B6, hCAR, C/EBPα, HNF4α, HNF3β were measured using real-time RT-PCR assays as detailed in the <i>Materials and Methods</i>. Relative gene expression levels from all donors were normalized against a randomly selected single donor. Leaner regression between CYP2B6 and one of these hepatic transcriptional factors was analyzed individually using Pearson’s Correlation Coefficient (JMP 7.0; SAS, NC).</p

Transactivation of CYP2B6 5’-flanking reporter constructs by HNF3β.

<p>A computer-based search for the first 2 kb of CYP2B6 upstream resulted in the identification of three potential HNF3β-responsive elements located between -1893bp and -350bp (A). HepG2 cells were transfected with HNF3β expression vector in the presence of CYP2B6 promoter constructs containing sequential deletion fragments (B) or the CYP2B6-2.0k harboring one of the mutated HNF3β binding sites (C). Forty eight hours post-transfection, luciferase activities were determined and expressed relative to the control (pGL3-Basic). Data represent the mean ± SD. (n = 3). (<i>*</i>, <i>p <0</i>.<i>05; **</i>, <i>p<0</i>.<i>01</i>).</p

Recruitment of HNF3β to enhancers identified upstream of CYP2B6 promoter.

<p>As detailed in the <i>Materials and Methods</i>, ChIP assays were used to analyze binding of HNF3β to the HNF3β-a (distal), HNF3β-c (proximal) containing, and the -1.6/-1.4kb regions in cultured HPH (A). After precipitation with HNF3β antibody, de-crosslinked DNA fragments were amplified by PCR. Amplification of the promoter of SULT1E1 was used as negative control as reported previously. In separate experiments, BIACORE SPR affinity assays (B) and (C) were carried out to measure the comparative binding kinetics of CYP2B6 enhancers on HNF3β as described under <i>Materials and Methods</i>. Sensorgrams of the interaction generated by the instrument were analyzed by the software BIAeval 3.2.</p

Effects of HNF3β on hCAR-mediated CYP2B6 activation in HepG2 cells.

<p>HepG2 cells were co-transfected with expression plasmids of hCAR, HNF3β or C/EBPα in the presence (A) or absence (B) of CYP2B6-2kb reporter construct as detailed in the <i>Materials and Methods</i>. Transfected cells were then treated with PB (1mM) and CITCO (1 μM) for 24 h. Dual luciferase activities (A) and CYP2B6 mRNA expression (B) were detected and expressed relative to vehicle control by reporter assay and real-time PCR analysis.</p

Effects of HNF3β on the expression and activity of CYP2B6 in HepG2 cells.

<p>HepG2 cells were infected with Negative Control Adenovirus (Ad-NC) or various amounts of adenovirus expressing HNF3β (Ad-HNF3β) for 48 h. Expression of HNF3β mRNA (A), CYP2B6 mRNA (B), and their proteins (C) were measured using real-time PCR and Western blotting assays, respectively. CYP2B6 enzymatic activity (D) was detected using P450-Glo^™ CYP2B6 Assay kit (Promega). In separate experiments, HepG2 cells were infected with pGreen Negtive Control Lentivirus (pGreen-NC) or HNF3β-RNAi lentivirus (HNF3β-shRNA) for 96 h before measuring the mRNA expression of HNF3β (E) and CYP2B6 (F) by real-time PCR. Results are expressed as the mean ± S.D. (n = 3). (<i>*P <0</i>.<i>05</i>, <i>**P <0</i>.<i>01</i>).</p

Controlled Protein Delivery from Electrospun Non-Wovens: Novel Combination of Protein Crystals and a Biodegradable Release Matrix

Poly-ε-caprolactone (PCL) is an excellent polymer for electrospinning and matrix-controlled drug delivery combining optimal processability and good biocompatibility. Electrospinning of proteins has been shown to be challenging via the use of organic solvents, frequently resulting in protein unfolding or aggregation. Encapsulation of protein crystals represents an attractive but largely unexplored alternative to established protein encapsulation techniques because of increased thermodynamic stability and improved solvent resistance of the crystalline state. We herein explore the electrospinning of protein crystal suspensions and establish basic design principles for this novel type of protein delivery system. PCL was deployed as a matrix, and lysozyme was used as a crystallizing model protein. By rational combination of lysozyme crystals 0.7 or 2.1 μm in diameter and a PCL fiber diameter between 1.6 and 10 μm, release within the first 24 h could be varied between approximately 10 and 100%. Lysozyme loading of PCL microfibers between 0.5 and 5% was achieved without affecting processability. While relative release was unaffected by loading percentage, the amount of lysozyme released could be tailored. PCL was blended with poly­(ethylene glycol) and poly­(lactic-<i>co</i>-glycolic acid) to further modify the release rate. Under optimized conditions, an almost constant lysozyme release over 11 weeks was achieved

Supplementary document for Plug-and-Play algorithm for imaging through scattering media - 6288232.pdf

1. The preprocessing procedure of PnPGAP-FPR 2. The optical memory effect (OME) range 3. Intermediate results of PnPGAP-FPR 4. Reconstruction results in real laboratory environmen

High Ionic Conductivity with Low Degradation in A‑Site Strontium-Doped Nonstoichiometric Sodium Bismuth Titanate Perovskite

High Ionic Conductivity with Low Degradation in A‑Site Strontium-Doped Nonstoichiometric Sodium Bismuth Titanate Perovskit

Localized and Whole-Room Effects of Portable Air Filtration Units on Aerosol Particle Deposition and Concentration in a Classroom Environment

In indoor environments with limited ventilation, recirculating portable air filtration (PAF) units may reduce COVID-19 infection risk via not only the direct aerosol route (i.e., inhalation) but also via an indirect aerosol route (i.e., contact with the surface where aerosol particles deposited). We systematically investigated the impact of PAF units in a mock classroom, as a supplement to background ventilation, on localized and whole-room surface deposition and particle concentration. Fluorescently tagged particles with a volumetric mean diameter near 2 μm were continuously introduced into the classroom environment via a breathing simulator with a prescribed inhalation–exhalation waveform. Deposition velocities were inferred on >50 horizontal and vertical surfaces throughout the classroom, while aerosol concentrations were spatially monitored via optical particle spectrometry. Results revealed a particle decay rate consistent with expectations based upon the reported clean air delivery rates of the PAF units. Additionally, the PAF units reduced peak concentrations by a factor of around 2.5 compared to the highest concentrations observed and led to a statistically significant reduction in deposition velocities for horizontal surfaces >2.5 m from the aerosol source. Our results not only confirm that PAF units can reduce particle concentrations but also demonstrate that they may lead to reduced particle deposition throughout an indoor environment when properly positioned with respect to the location of the particle source(s) within the room (e.g., where the largest group of students sit) and the predominant air distribution profile of the room

Aqueous Cold Sintering of Li-Based Compounds

Aqueous cold sintering of two lithium-based compounds, the electrolyte Li6.25La3Zr2Al0.25O12 (LLZAO) and cathode material LiCoO2 (LCO), is reported. For LLZAO, a relative density of ∼87% was achieved, whereas LCO was sintered to ∼95% with 20 wt % LLZAO as a flux/binder. As-cold sintered LLZAO exhibited a low total conductivity (10–8 S/cm) attributed to an insulating grain boundary blocking layer of Li2CO3. The blocking layer was reduced with a post-annealing process or, more effectively, by replacing deionized water with 5 M LiCl during cold sintering to achieve a total conductivity of ∼3 × 10–5 S/cm (similar to the bulk conductivity). For LCO-LLZAO composites, scanning electron microscopy and X-ray computer tomography indicated a continuous LCO matrix with the LLZAO phase evenly distributed but isolated throughout the ceramics. [001] texturing during cold sintering resulted in an order of magnitude difference in electronic conductivity between directions perpendicular and parallel to the c-axis at room temperature. The electronic conductivity (∼10–2 S/cm) of cold sintered LCO-LLZAO ceramics at room temperature was comparable to that of single crystals and higher than those synthesized via either conventional sintering or hot pressing