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https://www.readbyqxmd.com/read/27911376/rapid-and-efficient-generation-of-recombinant-human-pluripotent-stem-cells-by-recombinase-mediated-cassette-exchange-in-the-aavs1-locus
#1
Laura Ordovás, Ruben Boon, Mariaelena Pistoni, Yemiao Chen, Rangarajan Sambathkumar, Nicky Helsen, Jolien Vanhove, Pieter Berckmans, Qing Cai, Kim Vanuytsel, Susanna Raitano, Catherine M Verfaillie
Even with the revolution of gene-targeting technologies led by CRISPR-Cas9, genetic modification of human pluripotent stem cells (hPSCs) is still time consuming. Comparative studies that use recombinant lines with transgenes integrated into safe harbor loci could benefit from approaches that use site-specific targeted recombinases, like Cre or FLPe, which are more rapid and less prone to off-target effects. Such methods have been described, although they do not significantly outperform gene targeting in most aspects...
November 20, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911359/directed-protein-packaging-within-outer-membrane-vesicles-from-escherichia-coli-design-production-and-purification
#2
Nathan J Alves, Kendrick B Turner, Scott A Walper
An increasing interest in applying synthetic biology techniques to program outer membrane vesicles (OMV) are leading to some very interesting and unique applications for OMV where traditional nanoparticles are proving too difficult to synthesize. To date, all Gram-negative bacteria have been shown to produce OMV demonstrating packaging of a variety of cargo that includes small molecules, peptides, proteins and genetic material. Based on their diverse cargo, OMV are implicated in many biological processes ranging from cell-cell communication to gene transfer and delivery of virulence factors depending upon which bacteria are producing the OMV...
November 16, 2016: Journal of Visualized Experiments: JoVE
https://www.readbyqxmd.com/read/27911032/a-dermal-equivalent-engineered-with-tgf-%C3%AE-3-expressing-bone-marrow-stromal-cells-and-amniotic-membrane-cosmetic-healing-of-full-thickness-skin-wounds-in-rats
#3
Ali Samadikuchaksaraei, Ahmad Mehdipour, Mehryar Habibi Roudkenar, Javad Verdi, Mohammad Taghi Joghataei, Kamran As'adi, Fatemeh Amiri, Mozhgan Dehghan Harati, Mazaher Gholipourmalekabadi, Nushin Karkuki Osguei
Transforming growth factor beta-3 (TGF-β3) has been shown to decrease scar formation after scheduled topical applications to the cutaneous wounds. This study aimed to continuously deliver TGF-β3, during the early phase of wound healing, by engineering a dermal equivalent (DE) using TGF-β3 expressing bone marrow stromal cells (BM-SCs) and human dehydrated amniotic membrane (hDAM). To engineer a DE, rat BM-SCs were seeded on the hDAM and TGF-β3 was transiently transfected into the BM-SCs using a plasmid vector...
December 2016: Artificial Organs
https://www.readbyqxmd.com/read/27911010/screening-of-mercury-resistant-and-indole-3-acetic-acid-producing-bacterial-consortium-for-growth-promotion-of-cicer-arietinum-l
#4
Aatif Amin, Zakia Latif
Mercury resistant (Hg(R) ) bacteria were screened from industrial effluents and effluents-polluted rhizosphere soils near to districts Kasur and Sheikhupura, Pakistan. Out of 60 isolates, three bacterial strains, Bacillus sp. AZ-1, Bacillus cereus AZ-2, and Enterobacter cloacae AZ-3 showed Hg-resistance as 20 μg ml(-1) of HgCl2 and indole-3-acetic acid (IAA) production as 8-38 μg ml(-1) . Biochemical and molecular characterization of selected bacteria was confirmed by 16S ribotyping. Mercury resistant genes merA, merB, and merE of mer operon in Bacillus spp...
December 2, 2016: Journal of Basic Microbiology
https://www.readbyqxmd.com/read/27910896/methylmercury-induces-the-expression-of-tnf-%C3%AE-selectively-in-the-brain-of-mice
#5
Miyuki Iwai-Shimada, Tsutomu Takahashi, Min-Seok Kim, Masatake Fujimura, Hitoyasu Ito, Takashi Toyama, Akira Naganuma, Gi-Wook Hwang
Methylmercury selectively damages the central nervous system (CNS). The tumor necrosis factor (TNF) superfamily includes representative cytokines that participate in the inflammatory response as well as cell survival, and apoptosis. In this study, we found that administration of methylmercury selectively induced TNF-α expression in the brain of mice. Although the accumulated mercury concentration in the liver and kidneys was greater than in the brain, TNF-α expression was induced to a greater extent in brain...
December 2, 2016: Scientific Reports
https://www.readbyqxmd.com/read/27910729/identification-of-cytochrome-p450s-involved-in-the-metabolism-of-6-benzyl-1-benzyloxymethyl-5-iodouracil-w-1-using-human-recombinant-enzymes-and-rat-liver-microsomes-in-vitro
#6
Ying-Yuan Lu, Hai-Xu Cheng, Xin Wang, Xiao-Wei Wang, Jun-Yi Liu, Pu Li, Ya-Qing Lou, Jun Li, Chuang Lu, Guo-Liang Zhang
1. The aim of this study was to identify the hepatic metabolic enzymes, which involved in the biotransformation of 6-benzyl-1-benzyloxymethyl-5-iodouracil (W-1), a novel non-nucleoside reverse transcriptase inhibitor (NNRTIs) in rat and human in vitro. 2. The parent drug of W-1 was incubated with RLMs or recombinant CYPs (CYP1A2, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, CYP3A4 and CYP3A5, respectively) in the presence or absence of NADPH regenerating system. The metabolites of W-1 were analyzed with liquid chromatography-ion trap-time of flight-mass spectrometry (LC-IT-TOF-MS)...
December 2, 2016: Xenobiotica; the Fate of Foreign Compounds in Biological Systems
https://www.readbyqxmd.com/read/27910690/upgrading-a-high-throughput-spectrometer-for-high-frequency-400-khz-measurements
#7
T Nishizawa, M D Nornberg, D J Den Hartog, D Craig
The upgraded spectrometer used for charge exchange recombination spectroscopy on the Madison Symmetric Torus resolves emission fluctuations up to 400 kHz. The transimpedance amplifier's cutoff frequency was increased based upon simulations comparing the change in the measured photon counts for time-dynamic signals. We modeled each signal-processing stage of the diagnostic and scanned the filtering frequency to quantify the uncertainty in the photon counting rate. This modeling showed that uncertainties can be calculated based on assuming each amplification stage is a Poisson process and by calibrating the photon counting rate with a DC light source to address additional variation...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910689/high-resolution-main-ion-charge-exchange-spectroscopy-in-the-diii-d-h-mode-pedestal
#8
B A Grierson, K H Burrell, C Chrystal, R J Groebner, S R Haskey, D H Kaplan
A new high spatial resolution main-ion (deuterium) charge-exchange spectroscopy system covering the tokamak boundary region has been installed on the DIII-D tokamak. Sixteen new edge main-ion charge-exchange recombination sightlines have been combined with nineteen impurity sightlines in a tangentially viewing geometry on the DIII-D midplane with an interleaving design that achieves 8 mm inter-channel radial resolution for detailed profiles of main-ion temperature, velocity, charge-exchange emission, and neutral beam emission...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910672/improved-spatial-calibration-for-the-cxrs-system-on-east
#9
X H Yin, Y Y Li, J Fu, D Jiang, S Y Feng, Y Q Gu, Y Cheng, B Lyu, Y J Shi, M Y Ye, B N Wan
A Charge eXchange Recombination Spectroscopy (CXRS) diagnostic system has been developed to measure profiles of ion temperature and rotation since 2014 on EAST. Several techniques have been developed to improve the spatial calibration of the CXRS diagnostic. The sightline location was obtained by measuring the coordinates of three points on each sightline using an articulated flexible coordinate measuring arm when the vessel was accessible. After vacuum pumping, the effect of pressure change in the vacuum vessel was evaluated by observing the movement of the light spot from back-illuminated sightlines on the first wall using the newly developed articulated inspection arm...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910605/fast-ion-d%C3%AE-spectrum-diagnostic-in-the-east
#10
Y M Hou, C R Wu, J Huang, W W Heidbrink, M G von Hellermann, Z Xu, Z Jin, J F Chang, Y B Zhu, W Gao, Y J Chen, B Lyu, R J Hu, P F Zhang, L Zhang, W Gao, Z W Wu, Y Yu, M Y Ye
In toroidal magnetic fusion devices, fast-ion D-alpha diagnostic (FIDA) is a powerful method to study the fast-ion feature. The fast-ion characteristics can be inferred from the Doppler shifted spectrum of Dα light according to charge exchange recombination process between fast ions and probe beam. Since conceptual design presented in the last HTPD conference, significant progress has been made to apply FIDA systems on the Experimental Advanced Superconducting Tokamak (EAST). Both co-current and counter-current neutral beam injectors are available, and each can deliver 2-4 MW beam power with 50-80 keV beam energy...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910523/first-measurement-of-the-edge-charge-exchange-recombination-spectroscopy-on-east-tokamak
#11
Y Y Li, X H Yin, J Fu, D Jiang, S Y Feng, B Lyu, Y J Shi, Y Yi, X J Zhou, C D Hu, M Y Ye, B N Wan
An edge toroidal charge exchange recombination spectroscopy (eCXRS) diagnostic, based on a heating neutral beam injection (NBI), has been deployed recently on the Experimental Advanced Superconducting Tokamak (EAST). The eCXRS, which aims to measure the plasma ion temperature and toroidal rotation velocity in the edge region simultaneously, is a complement to the exiting core CXRS (cCXRS). Two rows with 32 fiber channels each cover a radial range from ∼2.15 m to ∼2.32 m with a high spatial resolution of ∼5-7 mm...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910459/upgrades-of-poloidal-and-tangential-x-ray-imaging-crystal-spectrometers-for-temperature-and-rotation-measurements-on-east
#12
Fudi Wang, Jun Chen, Ruiji Hu, Bo Lyu, Gilles Colledani, Jia Fu, Yingying Li, Manfred Bitter, Kenneth Hill, Sangon Lee, Minyou Ye, Yuejiang Shi, Baonian Wan
During the past two years, key parts of poloidal and tangential x-ray imaging crystal spectrometers (PXCSs and TXCSs) have been upgraded. For poloidal XCSs, double-crystals of ArXVII and FeXXV were deployed. For fulfilling in situ alignment of a poloidal XCS, the beryllium window must be flexibly removed. By utilizing a design, where the beryllium window was installed in the vacuum chamber of the double-crystal, and between the double-crystal and wall of this chamber, an in situ alignment for the two spectrometers was fulfilled...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910369/improved-edge-charge-exchange-recombination-spectroscopy-in-diii-d
#13
C Chrystal, K H Burrell, B A Grierson, S R Haskey, R J Groebner, D H Kaplan, A Briesemeister
The charge exchange recombination spectroscopy diagnostic on the DIII-D tokamak has been upgraded with the addition of more high radial resolution view chords near the edge of the plasma (r/a > 0.8). The additional views are diagnosed with the same number of spectrometers by placing fiber optics side-by-side at the spectrometer entrance with a precise separation that avoids wavelength shifted crosstalk without the use of bandpass filters. The new views improve measurement of edge impurity parameters in steep gradient, H-mode plasmas with many different shapes...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910350/optimization-and-application-of-cooled-avalanche-photodiodes-for-spectroscopic-fluctuation-measurements-with-ultra-fast-charge-exchange-recombination-spectroscopy
#14
D D Truong, R J Fonck, G R McKee
The Ultra-Fast Charge Exchange Recombination Spectroscopy (UF-CHERS) diagnostic is a highly specialized spectroscopic instrument with 2 spatial channels consisting of 8 spectral channels each and a resolution of ∼0.25 nm deployed at DIII-D to measure turbulent ion temperature fluctuations. Charge exchange emissions are obtained between 528 and 530 nm with 1 μs time resolution to study plasma instabilities. A primary challenge of extracting fluctuation measurements from raw UF-CHERS signals is photon and electronic noise...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910328/measurement-of-deuterium-density-profiles-in-the-h-mode-steep-gradient-region-using-charge-exchange-recombination-spectroscopy-on-diii-d
#15
S R Haskey, B A Grierson, K H Burrell, C Chrystal, R J Groebner, D H Kaplan, N A Pablant, L Stagner
Recent completion of a thirty two channel main-ion (deuterium) charge exchange recombination spectroscopy (CER) diagnostic on the DIII-D tokamak [J. L. Luxon, Nucl. Fusion 42, 614 (2002)] enables detailed comparisons between impurity and main-ion temperature, density, and toroidal rotation. In a H-mode DIII-D discharge, these new measurement capabilities are used to provide the deuterium density profile, demonstrate the importance of profile alignment between Thomson scattering and CER diagnostics, and aid in determining the electron temperature at the separatrix...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910316/measurement-of-the-deuterium-balmer-series-line-emission-on-east
#16
C R Wu, J Huang, W Gao, W Gao, Z Xu, J F Chang, Y M Hou, Z Jin, J C Xu, Y M Duan, P F Zhang, Y J Chen, L Zhang, Z W Wu, J G Li
Volume recombination plays an important role towards plasma detachment for magnetically confined fusion devices. High quantum number states of the Balmer series of deuterium are used to study recombination. On EAST (Experimental Advanced Superconducting Tokamak), two visible spectroscopic measurements are applied for the upper/lower divertor with 13 channels, respectively. Both systems are coupled with Princeton Instruments ProEM EMCCD 1024B camera: one is equipped on an Acton SP2750 spectrometer, which has a high spectral resolution ∼0...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910313/ion-temperature-and-toroidal-rotation-in-jet-s-low-torque-plasmas
#17
J Bernardo, M F F Nave, C Giroud, S Reyes Cortes, João P S Bizarro
This paper reports on the procedure developed as the best method to provide an accurate and reliable estimation of the ion temperature Ti and the toroidal velocity vϕ from Charge-eXchange Recombination Spectroscopy (CXRS) data from intrinsic rotation experiments at the Joint European Torus with the carbon wall. The low impurity content observed in such plasmas, resulting in low active CXRS signal, alongside low Doppler shifts makes the determination of Ti and vϕ particularly difficult. The beam modulation method will be discussed along with the measures taken to increase photon statistics and minimise errors from the absolute calibration and magneto-hydro-dynamics effects that may impact the CXRS passive emission...
November 2016: Review of Scientific Instruments
https://www.readbyqxmd.com/read/27910110/runs-of-homozygosity-current-knowledge-and-applications-in-livestock
#18
REVIEW
E Peripolli, D P Munari, M V G B Silva, A L F Lima, R Irgang, F Baldi
This review presents a broader approach to the implementation and study of runs of homozygosity (ROH) in animal populations, focusing on identifying and characterizing ROH and their practical implications. ROH are continuous homozygous segments that are common in individuals and populations. The ability of these homozygous segments to give insight into a population's genetic events makes them a useful tool that can provide information about the demographic evolution of a population over time. Furthermore, ROH provide useful information about the genetic relatedness among individuals, helping to minimize the inbreeding rate and also helping to expose deleterious variants in the genome...
December 1, 2016: Animal Genetics
https://www.readbyqxmd.com/read/27910048/gene-transfer-in-isolated-adult-cardiomyocytes
#19
Kjetil Hodne, David B Lipsett, William E Louch
During the past few decades, gene delivery using recombinant virus has made tremendous progress. With a higher than 80 % transduction efficiency, even in non-dividing cells, viral transduction has become the method of choice for efficient gene transfer into cardiomyocytes. However, in vitro gene delivery is dependent on a robust cell isolation protocol, as prolonged cultivation is needed to initiate gene expression and target specific cellular processes. This chapter describes some of the important steps that need to be considered for successful in vitro gene transfer into adult cardiomyocytes...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/27910043/production-and-characterization-of-vectors-based-on-the-cardiotropic-aav-serotype-9
#20
Erik Kohlbrenner, Thomas Weber
Vectors based on adeno-associated virus serotype 9 (AAV9) efficiently transduce cardiomyocytes in both rodents and large animal models upon either systemic or regional vector delivery. In this chapter, we describe the most widely used production and purification method of AAV9. This production approach does not depend on the use of a helpervirus but instead on transient transfection of HEK293T cells with a plasmid containing the recombinant AAV genome and a second plasmid encoding the AAV9 capsid proteins, the AAV Rep proteins and the adenoviral helper functions...
2017: Methods in Molecular Biology
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