Characterization of a panel of six β2-adrenergic receptor antibodies by indirect immunofluorescence microscopy

<p>Abstract</p> <p>Background</p> <p>The β₂-adrenergic receptor (β₂AR) is a primary target for medications used to treat asthma. Due to the low abundance of β₂AR, very few studies have reported its localization in tissues. However, the intracellular location of β₂AR in lung tissue, especially in airway smooth muscle cells, is very likely to have a significant impact on how the airways respond to β-agonist medications. Thus, a method for visualizing β₂AR in tissues would be of utility. The purpose of this study was to develop an immunofluorescent labeling technique for localizing native and recombinant β₂AR in primary cell cultures.</p> <p>Methods</p> <p>A panel of six different antibodies were evaluated in indirect immunofluorescence assays for their ability to recognize human and rat β₂AR expressed in HEK 293 cells. Antibodies capable of recognizing rat β₂AR were identified and used to localize native β₂AR in primary cultures of rat airway smooth muscle and epithelial cells. β₂AR expression was confirmed by performing ligand binding assays using the β-adrenergic antagonist [3H] dihydroalprenolol ^([3H]DHA).</p> <p>Results</p> <p>Among the six antibodies tested, we identified three of interest. An antibody developed against the C-terminal 15 amino acids of the human β₂AR (Ab-Bethyl) specifically recognized human but not rat β₂AR. An antibody developed against the C-terminal domain of the mouse β₂AR (Ab-sc570) specifically recognized rat but not human β₂AR. An antibody developed against 78 amino acids of the C-terminus of the human β₂AR (Ab-13989) was capable of recognizing both rat and human β₂ARs. In HEK 293 cells, the receptors were predominantly localized to the cell surface. By contrast, about half of the native rat β₂AR that we visualized in primary cultures of rat airway epithelial and smooth muscle cells using Ab-sc570 and Ab-13989 was found inside cells rather than on their surface.</p> <p>Conclusion</p> <p>Antibodies have been identified that recognize human β₂AR, rat β₂AR or both rat and human β₂AR. Interestingly, the pattern of expression in transfected cells expressing millions of receptors was dramatically different from that in primary cell cultures expressing only a few thousand native receptors. We anticipate that these antibodies will provide a valuable tool for evaluating the expression and trafficking of β₂AR in tissues.</p

Comparing Galaxy Morphology at Ultraviolet and Optical Wavelengths

We have undertaken an imaging survey of 34 nearby galaxies in far-ultraviolet (FUV, ~1500A) and optical (UBVRI) passbands to characterize galaxy morphology as a function of wavelength. This sample, which includes a range of classical Hubble types from elliptical to irregular with emphasis on spirals at low inclination angle, provides a valuable database for comparison with images of high-z galaxies whose FUV light is redshifted into the optical and near- infrared bands. Ultraviolet data are from the UIT Astro-2 mission. We present images and surface brightness profiles for each galaxy, and we discuss the wavelength-dependence of morphology for different Hubble types in the context of understanding high-z objects. In general, the dominance of young stars in the FUV produces the patchy appearance of a morphological type later than that inferred from optical images. Prominent rings and circumnuclear star formation regions are clearly evident in FUV images of spirals, while bulges, bars, and old, red stellar disks are faint to invisible at these short wavelengths. However, the magnitude of the change in apparent morphology ranges from dramatic in early--type spirals with prominent optical bulges to slight in late-type spirals and irregulars, in which young stars dominate both the UV and optical emission. Starburst galaxies with centrally concentrated, symmetric bursts display an apparent ``E/S0'' structure in the FUV, while starbursts associated with rings or mergers produce a peculiar morphology. We briefly discuss the inadequacy of the optically-defined Hubble sequence to describe FUV galaxy images and estimate morphological k-corrections, and we suggest some directions for future research with this dataset.Comment: Accepted for publication in the ApJS. 15 pages, 17 JPEG figures, 10 GIF figures. Paper and full resolution figures available at http://nedwww.ipac.caltech.edu/level5/Kuchinski/frames.htm

Prediction of photoperiodic regulators from quantitative gene circuit models

Photoperiod sensors allow physiological adaptation to the changing seasons. The external coincidence hypothesis postulates that a light-responsive regulator is modulated by a circadian rhythm. Sufficient data are available to test this quantitatively in plants, though not yet in animals. In Arabidopsis, the clock-regulated genes CONSTANS (CO) and FLAVIN, KELCH, F-BOX (FKF1) and their lightsensitive proteins are thought to form an external coincidence sensor. We use 40 timeseries of molecular data to model the integration of light and timing information by CO, its target gene FLOWERING LOCUS T (FT), and the circadian clock. Among other predictions, the models show that FKF1 activates FT. We demonstrate experimentally that this effect is independent of the known activation of CO by FKF1, thus we locate a major, novel controller of photoperiodism. External coincidence is part of a complex photoperiod sensor: modelling makes this complexity explicit and may thus contribute to crop improvement

Ultraviolet Imaging of the z=0.23 Cluster Abell 2246

We present deep ultraviolet observations of a field containing the cluster Abell 2246 (z=0.225) which provide far-ultraviolet (FUV) images of some of the faintest galaxies yet observed in that bandpass. Abell 2246 lies within the field of view of Ultraviolet Imaging Telescope (UIT) observations of the quasar HS1700+64, which accumulated over 7100 seconds of UIT FUV exposure time during the Astro-2 mission in March 1995. For objects found on both the FUV and ground-based V-band images, we obtain FUV (l ~ 1520 A) photometry and V-band photometry, as well as mid-UV (l ~ 2490 A) photometry from UIT Astro-1 observations and ground-based I-band photometry. We find five objects in the images which are probably galaxies at the distance of Abell 2246, with FUV magnitudes (m(FUV)) between 18.6 and 19.6, and V magnitudes between 18.4 and 19.6. We find that their absolute FUV fluxes and colors imply strongly that they are luminous galaxies with significant current star formation, as well as some relatively recent, but not current, (> 400 Myr ago) star formation. We interpret the colors of these five objects by comparing them with local objects, redshift-corrected template spectra and stellar population models, finding that they are plausibly matched by 10-Gyr-old population models with decaying star formation, with decay time constants in the range 3 Gyr < t < 5 Gyr, with an additional color component from a single burst of moderate ( ~ 400-500 Myr) age. From derived FUV luminosities we compute current star formation rates. We compare the UV properties of Abell 2246 with those of the Coma cluster, finding that Abell 2246 has significantly more recent star formation, consistent with the Butcher-Oemler phenomenon.Comment: Accepted for publication in the Astronomical Journal, June 1998. 17 Pages AAS latex, includes 4 bitmap .jpg format images and 4 other figures. PDF, Embedded Gzipped PS version (1.9Mb) TeX source and figures available at http://www.astro.virginia.edu/~bd4r/galaxies.htm

Shower development of particles with momenta from 15 GeV to 150 GeV in the CALICE scintillator-tungsten hadronic calorimeter

We present a study of showers initiated by electrons, pions, kaons, and protons with momenta from 15 GeV to 150 GeV in the highly granular CALICE scintillator-tungsten analogue hadronic calorimeter. The data were recorded at the CERN Super Proton Synchrotron in 2011. The analysis includes measurements of the calorimeter response to each particle type as well as measurements of the energy resolution and studies of the longitudinal and radial shower development for selected particles. The results are compared to Geant4 simulations (version 9.6.p02). In the study of the energy resolution we include previously published data with beam momenta from 1 GeV to 10 GeV recorded at the CERN Proton Synchrotron in 2010.Comment: 35 pages, 21 figures, 8 table

Pion and proton showers in the CALICE scintillator-steel analogue hadron calorimeter

Showers produced by positive hadrons in the highly granular CALICE scintillator-steel analogue hadron calorimeter were studied. The experimental data were collected at CERN and FNAL for single particles with initial momenta from 10 to 80 GeV/c. The calorimeter response and resolution and spatial characteristics of shower development for proton- and pion-induced showers for test beam data and simulations using Geant4 version 9.6 are compared.Comment: 26 pages, 16 figures, JINST style, changes in the author list, typos corrected, new section added, figures regrouped. Accepted for publication in JINS

Performance of the first prototype of the CALICE scintillator strip electromagnetic calorimeter

A first prototype of a scintillator strip-based electromagnetic calorimeter was built, consisting of 26 layers of tungsten absorber plates interleaved with planes of 45x10x3 mm3 plastic scintillator strips. Data were collected using a positron test beam at DESY with momenta between 1 and 6 GeV/c. The prototype's performance is presented in terms of the linearity and resolution of the energy measurement. These results represent an important milestone in the development of highly granular calorimeters using scintillator strip technology. This technology is being developed for a future linear collider experiment, aiming at the precise measurement of jet energies using particle flow techniques

The Time Structure of Hadronic Showers in highly granular Calorimeters with Tungsten and Steel Absorbers

The intrinsic time structure of hadronic showers influences the timing capability and the required integration time of hadronic calorimeters in particle physics experiments, and depends on the active medium and on the absorber of the calorimeter. With the CALICE T3B experiment, a setup of 15 small plastic scintillator tiles read out with Silicon Photomultipliers, the time structure of showers is measured on a statistical basis with high spatial and temporal resolution in sampling calorimeters with tungsten and steel absorbers. The results are compared to GEANT4 (version 9.4 patch 03) simulations with different hadronic physics models. These comparisons demonstrate the importance of using high precision treatment of low-energy neutrons for tungsten absorbers, while an overall good agreement between data and simulations for all considered models is observed for steel.Comment: 24 pages including author list, 9 figures, published in JINS

Ultraviolet Signatures of Tidal Interaction in the Giant Spiral Galaxy, M101

We present new evidence for tidal interactions having occurred in the disk of M101 in the last 10^8 - 10^9 years. Recent imaging of the far-ultraviolet emission from M101 by the Shuttle-borne Ultraviolet Imaging Telescope (UIT) reveals with unprecedented clarity a disk-wide pattern of multiple linear arm segments (``crooked arms''). The deep FUV image also shows a faint outer spiral arm with a (``curly tail'') feature that appears to loop around the supergiant HII region NGC 5471 - linking this outlying starburst with the rest of the galaxy. These FUV-bright features most likely trace hot O & B-type stars along with scattered light from associated nebular dust. Counterparts of the outermost ``crooked arms'' are evident in maps at visible wavelengths and in the 21-cm line of HI. The inner-disk FUV arms are most closely associated with H$\alpha$ knots and the outer (downstream) sides of CO arms. Comparisons of the ``crooked arm'' and ``curly tail'' morphologies with dynamical simulations yield the greatest similitude, when the non- axisymmetric forcing comes from a combination of ``external interactions'' with one or more companion galaxies and ``internal perturbations'' from massive objects orbiting within the disk. We speculate that NGC 5471 represents one of these ``massive disturbers'' within the disk, whose formation followed from a tidal interaction between M101 and a smaller galaxy.Comment: Paper format (latex); length of paper (8); 4 gif figure files; uses aas2pp4.sty AASTeX macro file; to be published in Part I of the Astrophysical Journa

RNAi Screening in Drosophila Cells Identifies New Modifiers of Mutant Huntingtin Aggregation

The fruitfly Drosophila melanogaster is well established as a model system in the study of human neurodegenerative diseases. Utilizing RNAi, we have carried out a high-throughput screen for modifiers of aggregate formation in Drosophila larval CNS-derived cells expressing mutant human Huntingtin exon 1 fused to EGFP with an expanded polyglutamine repeat (62Q). 7200 genes, encompassing around 50% of the Drosophila genome, were screened, resulting in the identification of 404 candidates that either suppress or enhance aggregation. These candidates were subjected to secondary screening in normal length (18Q)-expressing cells and pruned to remove dsRNAs with greater than 10 off-target effects (OTEs). De novo RNAi probes were designed and synthesized for the remaining 68 candidates. Following a tertiary round of screening, 21 high confidence candidates were analyzed in vivo for their ability to modify mutant Huntingtin-induced eye degeneration and brain aggregation. We have established useful models for the study of human HD using the fly, and through our RNAi screen, we have identified new modifiers of mutant human Huntingtin aggregation and aggregate formation in the brain. Newly identified modifiers including genes related to nuclear transport, nucleotide processes, and signaling, may be involved in polyglutamine aggregate formation and Huntington disease cascades