High-throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

Nonviral gene delivery methods are advantageous over viral vectors in terms of safety, cost, and flexibility in design and application, but suffer from lower gene transfer efficiency. In addition to modifications to nucleic acid design and nonviral carriers, new tools are sought to enhance transfection. Priming is the pharmacological modulation of transfection efficiency and transgene expression, and has demonstrated transfection increase in several compounds, for example, chloroquine and glucocorticoids. To develop a library of transfection priming compounds, a highthroughput screen was performed of the NIH Clinical Collection (NCC) to identify clinical compounds that prime polyethylenimine (PEI) transfection. HEK293T cells were treated with priming compounds, then transfected with enhanced green fluorescent protein (EGFP)-encoding plasmid by PEI. After 48-hr culture, primed and transfected cells were assayed for transfection, cell proliferation, and cell viability by fluorescence measurement of EGFP reporter, Hoechst 33342 nuclei stain, and resazurin metabolic assay. From the microscope image analysis and microplate measurements, transfection fold-changes were determined, and compounds resulting in statistically significant transfection fold-change were identified. NCC compounds were clustered using PubChem fingerprint similarity by Tanimoto coefficients in ChemmineTools. Fold-changes for each compound were linked to drug clusters, from which drug classes that prime transfection were identified. Among the identified drugs classes that primed transfection increases were antioxidants, GABAA receptor modulators, and glucocorticoids. Resveratrol and piceid, stilbenoid antioxidants found in grapes, and zolpidem, a GABAA modulator, increased transfection nearly three-fold. Literature indicate interaction of the identified transfection priming drug clusters with mitochondria, which may modulate mitochondrial dysfunction known to be associated with PEI transfection

Adaptive thermal compensation of test masses in advanced LIGO

As the first generation of laser interferometric gravitational wave detectors near operation, research and development has begun on increasing the instrument's sensitivity while utilizing the existing infrastructure. In the Laser Interferometer Gravitational Wave Observatory (LIGO), significant improvements are being planned for installation in ~2007, increasing strain sensitivity through improved suspensions and test mass substrates, active seismic isolation, and higher input laser power. Even with the highest quality optics available today, however, finite absorption of laser power within transmissive optics, coupled with the tremendous amount of optical power circulating in various parts of the interferometer, result in critical wavefront deformations which would cripple the performance of the instrument. Discussed is a method of active wavefront correction via direct thermal actuation on optical elements of the interferometer. A simple nichrome heating element suspended off the face of an affected optic will, through radiative heating, remove the gross axisymmetric part of the original thermal distortion. A scanning heating laser will then be used to remove any remaining non-axisymmetric wavefront distortion, generated by inhomogeneities in the substrate's absorption, thermal conductivity, etc. A proof-of-principle experiment has been constructed at MIT, selected data of which are presented.Comment: 11 pages, 7 figures, submitted to Classical and Quantum Gravit

Characterization of Spatial Coherence of Synchrotron Radiation with Non-Redundant Arrays of Apertures

We present a method to characterize the spatial coherence of soft X-ray radiation from a single diffraction pattern. The technique is based on scattering from non-redundant arrays (NRA) of slits and records the degree of spatial coherence at several relative separations from one to 15 microns, simultaneously. Using NRAs we measured the transverse coherence of the X-ray beam at the XUV X-ray beamline P04 of the PETRA III synchrotron storage ring as a function of different beam parameters. To verify the results obtained with the NRAs additional Young's double pinhole experiments were conducted and show good agreement.Comment: 15 pages, 6 figures, 2 tables, 42 reference

Sensitivity limitations in optical speed meter topology of gravitational-wave antennae

The possible design of QND gravitational-wave detector based on speed meter principle is considered with respect to optical losses. The detailed analysis of speed meter interferometer is performed and the ultimate sensitivity that can be achieved is calculated. It is shown that unlike the position meter signal-recycling can hardly be implemented in speed meter topology to replace the arm cavities as it is done in signal-recycled detectors, such as GEO 600. It is also shown that speed meter can beat the Standard Quantum Limit (SQL) by the factor of $\sim 3$ in relatively wide frequency band, and by the factor of $\sim 10$ in narrow band. For wide band detection speed meter requires quite reasonable amount of circulating power $\sim 1$ MW. The advantage of the considered scheme is that it can be implemented with minimal changes in the current optical layout of LIGO interferometer.Comment: 20 pages, 12 figure

Observational evidence for the convective transport of dust over the central United States

Bulk aerosol composition and aerosol size distributions measured aboard the DC-8 aircraft during the Deep Convective Clouds and Chemistry Experiment mission in May/June 2012 were used to investigate the transport of mineral dust through nine storms encountered over Colorado and Oklahoma. Measurements made at low altitudes (\u3c5 km mean sea level (MSL)) in the storm inflow region were compared to those made in cirrus anvils (altitude \u3e 9 km MSL). Storm mean outflow Ca2+ mass concentrations and total coarse (1 µm \u3c diameter \u3c 5 µm) aerosol volume (Vc) were comparable to mean inflow values as demonstrated by average outflow/inflow ratios greater than 0.5. A positive relationship between Ca2+, Vc, ice water content, and large (diameter \u3e 50 µm) ice particle number concentrations was not evident; thus, the influence of ice shatter on these measurements was assumed small. Mean inflow aerosol number concentrations calculated over a diameter range (0.5 µm \u3c diameter \u3c 5.0 µm) relevant for proxy ice nuclei (NPIN) were ~15–300 times higher than ice particle concentrations for all storms. Ratios of predicted interstitial NPIN (calculated as the difference between inflow NPIN and ice particle concentrations) and inflow NPIN were consistent with those calculated for Ca2+ and Vc and indicated that on average less than 10% of the ingested NPIN were activated as ice nuclei during anvil formation. Deep convection may therefore represent an efficient transport mechanism for dust to the upper troposphere where these particles can function as ice nuclei cirrus forming in situ

High-throughput screening of clinically approved drugs that prime polyethylenimine transfection reveals modulation of mitochondria dysfunction response improves gene transfer efficiencies

Nonviral gene delivery methods are advantageous over viral vectors in terms of safety, cost, and flexibility in design and application, but suffer from lower gene transfer efficiency. In addition to modifications to nucleic acid design and nonviral carriers, new tools are sought to enhance transfection. Priming is the pharmacological modulation of transfection efficiency and transgene expression, and has demonstrated transfection increase in several compounds, for example, chloroquine and glucocorticoids. To develop a library of transfection priming compounds, a highthroughput screen was performed of the NIH Clinical Collection (NCC) to identify clinical compounds that prime polyethylenimine (PEI) transfection. HEK293T cells were treated with priming compounds, then transfected with enhanced green fluorescent protein (EGFP)-encoding plasmid by PEI. After 48-hr culture, primed and transfected cells were assayed for transfection, cell proliferation, and cell viability by fluorescence measurement of EGFP reporter, Hoechst 33342 nuclei stain, and resazurin metabolic assay. From the microscope image analysis and microplate measurements, transfection fold-changes were determined, and compounds resulting in statistically significant transfection fold-change were identified. NCC compounds were clustered using PubChem fingerprint similarity by Tanimoto coefficients in ChemmineTools. Fold-changes for each compound were linked to drug clusters, from which drug classes that prime transfection were identified. Among the identified drugs classes that primed transfection increases were antioxidants, GABAA receptor modulators, and glucocorticoids. Resveratrol and piceid, stilbenoid antioxidants found in grapes, and zolpidem, a GABAA modulator, increased transfection nearly three-fold. Literature indicate interaction of the identified transfection priming drug clusters with mitochondria, which may modulate mitochondrial dysfunction known to be associated with PEI transfection

HSRL-2 aerosol optical measurements and microphysical retrievals vs. airborne in situ measurements during DISCOVER-AQ 2013: : an intercomparison study

This is an Open Access article distributed under the Creative Commons Attribution 3.0 License, https://creativecommons.org/licenses/by/3.0/. © Author(s) 2017. Published by Copernicus Publications on behalf of the European Geosciences Union.We present a detailed evaluation of remotely-sensed aerosol microphysical properties obtained from an advanced, multi-wavelength High Spectral Resolution Lidar (HSRL-2) during the 2013 NASA DISCOVER-AQ field campaign. Vertically resolved retrievals of fine mode aerosol number, surface area, and volume concentration as well as aerosol effective radius are compared to 108 co-located, airborne in situ measurement profiles in the wintertime San Joaquin Valley, California, and in summertime Houston, Texas. An algorithm for relating the dry in situ aerosol properties to those obtained by the HSRL at ambient relative humidity is discussed. We show that the HSRL-2 retrievals of ambient fine mode aerosol surface area and volume concentrations agree with the in situ measurements to within 25% and 10%, respectively, once hygroscopic growth adjustments have been applied to the dry in situ data. Despite this excellent agreement for the microphysical properties, extinction and backscatter coefficients at ambient relative humidity derived from the in situ aerosol measurements using Mie theory are consistently smaller than those measured by the HSRL, with average differences of 31% 5% and 53% 11% for California and Texas, respectively. This low bias in the in situ estimates is attributed to the presence of coarse mode aerosol that are detected by HSRL-2 but that are too large to be well sampled by the in situ instrumentation. Since the retrieval of aerosol volume is most relevant to current regulatory efforts targeting fine particle mass (PM2:5), these findings highlight the advantages of an advanced 3+2 HSRL for constraining the vertical distribution of the aerosol volume or mass loading relevant for air quality.Peer reviewedFinal Published versio

QND measurements for future gravitational-wave detectors

Second-generation interferometric gravitational-wave detectors will be operating at the Standard Quantum Limit, a sensitivity limitation set by the trade off between measurement accuracy and quantum back action, which is governed by the Heisenberg Uncertainty Principle. We review several schemes that allows the quantum noise of interferometers to surpass the Standard Quantum Limit significantly over a broad frequency band. Such schemes may be an important component of the design of third-generation detectors.Comment: 22 pages, 6 figures, 1 table; In version 2, more tutorial information on quantum noise in GW interferometer and several new items into Reference list were adde