Radiographic diagnoses in 80 cats before and 73 cats after unobstructing the urethra.

OBJECTIVE: The aim of this study was to investigate causes for feline urethral obstruction and determine whether the frequency of radiographic diagnoses differs between cats radiographed before or after unobstruction of the urethra. MATERIALS AND METHODS: A retrospective cross-sectional study of cats with naturally occurring urethral obstruction was performed. Only cats presenting for their first urethral obstruction in which radiography was integrated in the initial evaluation were included. The diagnosis frequency (overall and for each disease type) was compared between cats radiographed before or after unobstruction of the urethra. RESULTS: Eighty cats (52%) had radiographs obtained before unobstructing the urethra and 73 cats (48%) had radiographs taken after unobstructing the urethra. Cats radiographed before unobstruction had a greater frequency of radiographic diagnoses than those radiographed after unobstruction (61% versus 45%). This difference was largely due to a greater frequency of urethral plugs detected before unobstruction versus after unobstruction (45% versus 5.5%). CLINICAL SIGNIFICANCE: Radiographs obtained before unobstructing the urethra provided a diagnostic advantage for detecting a cause for urethral obstruction compared to radiographs obtained after unobstructing the urethra. Urethral plugs were the most common diagnosis.

Solid Cherenkov detector for studying nucleosynthesis in inertial confinement fusion.

Measuring gamma rays emitted from nuclear reactions gives insight into their nuclear structure. Notably, there are several nuclear reactions that produce gamma rays at ∼1 MeV-3 MeV energies such as T(4He, γ)7Li, 4He(3He, γ)7Be, and 12C(p, γ)13N, which may solve questions lingering about big-bang nucleosynthesis and stellar nucleosynthesis. To observe 1 MeV-3 MeV gamma rays in an inertial confinement fusion system, a new style of the Cherenkov detector was developed using aerogel and fused silica as a Cherenkov medium. Utilizing the OMEGA laser facility, both aerogel and fused silica media were compared with the existing gas-medium Cherenkov detector to validate the concept. Gamma ray measurements from high yield inertial confinement fusion implosions (deuterium-tritium and deuterium-3He) demonstrated that aerogel and fused silica were viable Cherenkov media, paving the way for a potential optimized detector to make these cross section measurements on OMEGA or the National Ignition Facility.

Progress on next generation gamma-ray Cherenkov detectors for the National Ignition Facility.

Fusion reaction history and ablator areal density measurements for Inertial Confinement Fusion experiments at the National Ignition Facility are currently conducted using the Gamma Reaction History diagnostic (GRH_6m). Future Gas Cherenkov Detectors (GCDs) will ultimately provide ∼100x more sensitivity, reduce the effective temporal response from ∼100 to ∼10 ps, and lower the energy threshold from 2.9 to 1.8 MeV, relative to GRH_6m. The first phase toward next generation GCDs consisted of inserting the existing coaxial GCD-3 detector into a reentrant well which puts it within 4 m of the implosion. Reaction history and ablator gamma measurement results from this Phase I are discussed here. These results demonstrate viability for the follow-on Phases of (II) the use of a revolutionary new pulse-dilation photomultiplier tube to improve the effective measurement bandwidth by >10x relative to current PMT technology; and (III) the design of a NIF-specific "Super" GCD which will be informed by the assessment of the radiation background environment within the well described here.

Next generation gamma-ray Cherenkov detectors for the National Ignition Facility.

The newest generation of Gas Cherenkov Detector (GCD-3) employed in Inertial Confinement Fusion experiments at the Omega Laser Facility has provided improved performance over previous generations. Comparison of reaction histories measured using two different deuterium-tritium fusion products, namely gamma rays using GCD and neutrons using Neutron Temporal Diagnostic (NTD), have provided added credibility to both techniques. GCD-3 is now being brought to the National Ignition Facility (NIF) to supplement the existing Gamma Reaction History (GRH-6m) located 6 m from target chamber center (TCC). Initially it will be located in a reentrant well located 3.9 m from TCC. Data from GCD-3 will inform the design of a heavily-shielded "Super" GCD to be located as close as 20 cm from TCC. It will also provide a test-bed for faster optical detectors, potentially lowering the temporal resolution from the current ∼100 ps state-of-the-art photomultiplier tubes (PMT) to ∼10 ps Pulse Dilation PMT technology currently under development.

Aerogel Cherenkov detector for characterizing the intense flash x-ray source, Cygnus, spectrum.

An aerogel Cherenkov detector is proposed to measure the X-ray energy spectrum from the Cygnus-intense flash X-ray source operated at the Nevada National Security Site. An array of aerogels set at a variety of thresholds between 1 and 3 MeV will be adequate to map out the bremsstrahlung X-ray production of the Cygnus, where the maximum energy of the spectrum is normally around 2.5 MeV. In addition to the Cherenkov radiation from aerogels, one possible competing light-production mechanism is optical transition radiation (OTR), which may be significant in aerogels due to the large number of transitions from SiO2 clusters to vacuum voids. To examine whether OTR is a problem, four aerogel samples were tested using a mono-energetic electron beam (varied in the range of 1-3 MeV) at NSTec Los Alamos Operations. It was demonstrated that aerogels can be used as a Cherenkov medium, where the rate of the light production is about two orders magnitude higher when the electron beam energy is above threshold.

Conceptual design of the gamma-to-electron magnetic spectrometer for the National Ignition Facility.

The Gamma-to-Electron Magnetic Spectrometer (GEMS) diagnostic is designed to measure the prompt γ-ray energy spectrum during high yield deuterium-tritium (DT) implosions at the National Ignition Facility (NIF). The prompt γ-ray spectrum will provide "burn-averaged" observables, including total DT fusion yield, total areal density (ρR), ablator ρR, and fuel ρR. These burn-averaged observables are unique because they are essentially averaged over 4π, providing a global reference for the line-of-sight-specific measurements typical of x-ray and neutron diagnostics. The GEMS conceptual design meets the physics-based requirements: ΔE/E = 3%-5% can be achieved in the range of 2-25 MeV γ-ray energy. Minimum DT neutron yields required for 15% measurement uncertainty at low-resolution mode are: 5 × 10(14) DT-n for ablator ρR (at 0.2 g/cm(2)); 2 × 10(15) DT-n for total DT yield (at 4.2 × 10(-5) γ/n); and 1 × 10(16) DT-n for fuel ρR (at 1 g/cm(2)).

Extended performance gas Cherenkov detector for gamma-ray detection in high-energy density experiments.

A new Gas Cherenkov Detector (GCD) with low-energy threshold and high sensitivity, currently known as Super GCD (or GCD-3 at OMEGA), is being developed for use at the OMEGA Laser Facility and the National Ignition Facility (NIF). Super GCD is designed to be pressurized to ≤400 psi (absolute) and uses all metal seals to allow the use of fluorinated gases inside the target chamber. This will allow the gamma energy threshold to be run as low at 1.8 MeV with 400 psi (absolute) of C2F6, opening up a new portion of the gamma ray spectrum. Super GCD operating at 20 cm from TCC will be ∼400 × more efficient at detecting DT fusion gammas at 16.7 MeV than the Gamma Reaction History diagnostic at NIF (GRH-6m) when operated at their minimum thresholds.

Gamma-to-electron magnetic spectrometer (GEMS): an energy-resolved γ-ray diagnostic for the National Ignition Facility.

The gamma-to-electron magnetic spectrometer, having better than 5% energy resolution, is proposed to resolve γ-rays in the range of E(o) ± 20% in single shot, where E(o) is the central energy and is tunable from 2 to 25 MeV. Gamma-rays from inertial confinement fusion implosions interact with a thin Compton converter (e.g., beryllium) located at approximately 300 cm from the target chamber center (TCC). Scattered electrons out of the Compton converter enter an electromagnet placed outside the NIF chamber (approximately 600 cm from TCC) where energy selection takes place. The electromagnet provides tunable E(o) over a broad range in a compact manner. Energy resolved electrons are measured by an array of quartz Cherenkov converters coupled to photomultipliers. Given 100 detectable electrons in the energy bins of interest, 3 × 10(14) minimum deuterium/tritium (DT) neutrons will be required to measure the 4.44 MeV (12)C γ-rays assuming 200 mg/cm(2) plastic ablator areal density and 3 × 10(15) minimum DT neutrons to measure the 16.75 MeV DT γ-ray line.

Mach-Zehnder recording systems for pulsed power diagnostics.

Fiber-optic transmission and recording systems, based on Mach-Zehnder modulators, have been developed and installed at the National Ignition Facility (NIF), and are being developed for other pulsed-power facilities such as the Z accelerator at Sandia, with different requirements. We present the design and performance characteristics for the mature analog links, based on the system developed for the Gamma Reaction History diagnostic at the OMEGA laser and at NIF. For a single detector channel, two Mach-Zehnders are used to provide high dynamic range at the full recording bandwidth with no gaps in the coverage. We present laboratory and shot data to estimate upper limits on the radiation effects as they impact recorded data quality. Finally, we will assess the technology readiness level for mature and developing implementations of Mach-Zehnder links for these environments.

Development and characterization of sub-100 ps photomultiplier tubes.

We describe the evaluation of a microchannel plate (MCP) photomultiplier tube (PMT), incorporating a 3 µm pore MCP and constant voltage anode and cathode gaps. The use of the small pore size results in PMTs with response functions of the order of 85 ps full-width-half-maximum, while the constant electric field across the anode and cathode gaps produces a uniform response function over the entire operating range of the device. The PMT was characterized on a number of facilities and employed on gas Cherenkov detectors fielded on various deuterium tritium fuel (DT) implosions on the Omega Laser Facility at the University of Rochester. The Cherenkov detectors are part of diagnostic development to measure Gamma ray reaction history for DT implosions on the National Ignition Facility.

Gamma bang time analysis at OMEGA.

Absolute bang time measurements with the gas Cherenkov detector (GCD) and gamma reaction history (GRH) diagnostic have been performed to high precision at the OMEGA laser facility at the University of Rochester with bang time values for the two diagnostics agreeing to within 5 ps on average. X-ray timing measurements of laser-target coupling were used to calibrate a facility-generated laser timing fiducial with rms spreads in the measured coupling times of 9 ps for both GCD and GRH. Increased fusion yields at the National Ignition Facility (NIF) will allow for improved measurement precision with the GRH easily exceeding NIF system design requirements.

GEANT4 simulations of Cherenkov reaction history diagnostics.

This paper compares the results from a GEANT4 simulation of the gas Cherenkov detector 1 (GCD1) with previous simulations and experimental data from the Omega laser facility. The GCD1 collects gammas emitted during a deuterium-tritium capsule implosion and converts them, through several processes, to Cherenkov light. Photon signals are recorded using subnanosecond photomultiplier tubes, producing burn reaction histories. The GEANT4 GCD1 simulation is first benchmarked against ACCEPT, an integrated tiger series code, with good agreement. The simulation is subsequently compared with data from the Omega laser facility, where experiments have been performed to measure the effects of Hohlraum materials on reaction history signals, in preparation for experiments at the National Ignition Facility.

Using gamma-ray emission to measure areal density of inertial confinement fusion capsules.

Fusion neutrons streaming from a burning inertial confinement fusion capsule generate gamma rays via inelastic nuclear scattering in the ablator of the capsule. The intensity of gamma-ray emission is proportional to the product of the ablator areal density (ρR) and the yield of fusion neutrons, so by detecting the gamma rays we can infer the ablator areal density, provided we also have a measurement of the capsule's total neutron yield. In plastic-shell capsules, for example, (12)C nuclei emit gamma rays at 4.44 MeV after excitation by 14.1 MeV neutrons from D+T fusion. These gamma rays can be measured by a new gamma-ray detector under development. Analysis of predicted signals is in progress, with results to date indicating that the method promises to be useful for diagnosing imploded capsules.

Diagnosing inertial confinement fusion gamma ray physics (invited).

The gamma reaction history (GRH) diagnostic is a multichannel, time-resolved, energy-thresholded γ-ray spectrometer that provides a high-bandwidth, direct-measurement of fusion reaction history in inertial confinement fusion implosion experiments. 16.75 MeV deuterium+tritium (DT) fusion γ-rays, with a branching ratio of the order of 10(-5)γ/(14 MeV n), are detected to determine fundamental burn parameters, such as nuclear bang time and burn width, critical to achieving ignition at the National Ignition Facility. During the tritium/hydrogen/deuterium ignition tuning campaign, an additional γ-ray line at 19.8 MeV, produced by hydrogen+tritium fusion with a branching ratio of unity, will increase the available γ-ray signal and may allow measurement of reacting fuel composition or ion temperature. Ablator areal density measurements with the GRH are also made possible by detection of 4.43 MeV γ-rays produced by inelastic scatter of DT fusion neutrons on (12)C nuclei in the ablating plastic capsule material.

Release, biological potency, and biochemical integrity of recombinant human platelet-derived growth factor-BB (rhPDGF-BB) combined with Augment(TM) Bone Graft or GEM 21S beta-tricalcium phosphate (beta-TCP).

Over 10 million surgical procedures are performed annually in the United States to treat musculoskeletal injuries, and a significant portion of these involve orthopedic bone grafting. The goals of the study were to evaluate the in vitro and in vivo release kinetics, biological potency and biochemical integrity of rhPDGF-BB combined with large (1000-2000 microm) and small (250-1000 microm) beta-TCP particles. Recombinant human platelet-derived growth factor B homodimer (rhPDGF-BB) is a protein growth factor under development as a therapeutic for accelerating bone healing. Release of the protein was monitored in vitro by ELISA, and in vivo by measurement of radioactive rhPDGF-BB implanted in rat calvarial defects. Biological activity was measured using a cell-based bioassay, and biochemical integrity was determined by SDS-PAGE and high pressure size exclusion chromatography (HPSEC). Release of rhPDGF-BB occurred rapidly from beta-TCP both in vitro and in vivo. Almost 100% of the rhPDGF-BB was recovered from large and small beta-TCP after 90 min in vitro. Approximately 90% of the rhPDGF-BB was depleted from calvarial defect sites within 72 h of implantation. RhPDGF-BB retained 100% of its biological potency compared to reference standard rhPDGF-BB, manifested as a single band at ~30 kDa by SDS-PAGE and a single peak eluted after 13 min by HPSEC following release from beta-TCP. RhPDGF-BB is rapidly released from large and small beta-TCP particles and is biochemically unaltered following release.

Temporal regulation of the Mre11-Rad50-Nbs1 complex during adenovirus infection.

Adenovirus infection induces a cellular DNA damage response that can inhibit viral DNA replication and ligate viral genomes into concatemers. It is not clear if the input virus is sufficient to trigger this response or if viral DNA replication is required. Adenovirus has evolved two mechanisms that target the Mre11-Rad50-Nbs1 (MRN) complex to inhibit the DNA damage response. These include E4-ORF3-dependent relocalization of MRN proteins and E4-ORF6/E1B-55K-dependent degradation of MRN components. The literature suggests that degradation of the MRN complex due to E4-ORF6/E1B-55K does not occur until after viral DNA replication has begun. We show that, by the time viral DNA accumulates, the MRN complex is inactivated by either of the E4-induced mechanisms and that, with E4-ORF6/E1B-55K, this inactivation is due to MRN degradation. Our data are consistent with the conclusion that input viral DNA is sufficient to induce the DNA damage response. Further, we demonstrate that when the DNA damage response is active in E4 mutant virus infections, the covalently attached terminal protein is not cleaved from viral DNAs, and the viral origins of replication are not detectably degraded at a time corresponding to the onset of viral replication. The sequences of concatemeric junctions of viral DNAs were determined, which supports the conclusion that nonhomologous end joining mediates viral DNA ligation. Large deletions were found at these junctions, demonstrating nucleolytic procession of the viral DNA; however, the lack of terminal protein cleavage and terminus degradation at earlier times shows that viral genome deletion and concatenation are late effects.

Gamma bang time/reaction history diagnostics for the National Ignition Facility using 90 degrees off-axis parabolic mirrors.

Gas Cherenkov detectors (GCDs) have been used to convert fusion gamma into photons to achieve gamma bang time and reaction history measurements. The GCDs designed for OMEGA used Cassegrain reflector optics in order to fit inside a 10 in. manipulator. A novel design for the National Ignition Facility using 90 degrees off-axis parabolic mirrors will increase light collection efficiency from fusion gammas and achieve minimum time dispersion. The broadband Cherenkov light (from 200 to 800 nm) is relayed into a high-speed detector using three parabolic mirrors. Because light is collected from many source planes throughout the CO(2) gas volume, the detector is positioned at the stop position rather than at an image position. The stop diameter and its position are independent of the light-generation location along the gas cell. The current design collects light from a 100 mm diameter by 500 mm long gas volume. Optical ray tracings demonstrate how light can be collected from different angled trajectories of the Compton electrons as they fly through the CO(2) gas volume. A cluster of four channels will allow for increased dynamic range as well as for different gamma energy threshold sensitivities.

Granzyme H destroys the function of critical adenoviral proteins required for viral DNA replication and granzyme B inhibition.

Granzymes are key components of the immune response that play important roles in eliminating host cells infected by intracellular pathogens. Several granzymes are potent inducers of cell death. However, whether granzymes use additional mechanisms to exert their antipathogen activity remains elusive. Here, we show that in adenovirus-infected cells in which granzyme B (gzmB) and downstream apoptosis pathways are inhibited, granzyme H (gzmH), an orphan granzyme without known function, directly cleaves the adenovirus DNA-binding protein (DBP), a viral component absolutely required for viral DNA replication. We directly addressed the functional consequences of the cleavage of the DBP by gzmH through the generation of a virus that encodes a gzmH-resistant DBP. This virus demonstrated that gzmH directly induces an important decay in viral DNA replication. Interestingly, gzmH also cleaves the adenovirus 100K assembly protein, a major inhibitor of gzmB, and relieves gzmB inhibition. These results provide the first evidence that granzymes can mediate antiviral activity through direct cleavage of viral substrates, and further suggest that different granzymes have synergistic functions to outflank viral defenses that block host antiviral activities.

Catalase enrichment using recombinant adenovirus protects alphaTN4-1 cells from H(2)O(2).

Since oxidative stress has been implicated in the development of numerous diseases including cataract, this laboratory has created and investigated the stress response of murine immortal lens epithelial cell lines (alphaTN4-1) conditioned to withstand lethal peroxide concentrations. Two of a group of antioxidative defense (AOD) enzymes found in such cells to have markedly enhanced activity are catalase (CAT) and GSH S-transferase alpha2 (GST). In order to determine if enrichment of one or both of these AODs is sufficient to protect alphaTN4-1 cells from lethal H(2)O(2) levels, these cells were infected with adenovirus vectors capable of expressing these AODs at a high level. With this system, gene enrichment and increased enzyme activity were observed with both CAT and GST vectors. The percentage of cells infected ranged from about 50 to 90% depending on the multiplicity of infection (MOI). CAT but not GST protected the cells from H(2)O(2) stress. The CAT activity was increased from 15- to 150-fold and even at the lower levels protected the cells from H(2)O(2) concentrations as high as 200 microM or more (H(2)O(2) levels which rapidly kill non-enriched cells). Even when only about 50% of the cell population is infected as judged by GFP infection, the entire population appeared to be protected based on cell viability. The CAT enrichment appears to protect other intracellular defense systems such as GSH from being depleted in contrast to non-enriched cell populations where GSH is rapidly exhausted. The overall results suggest that enriching the cellular CAT gene level with an appropriate recombinant viral vector may be sufficient to protect in vivo systems from peroxide stress.

In situ detection of specific DNA double strand breaks using rolling circle amplification.

We have developed a method to localize DNA double strand breaks (DSBs) in situ in cultured mammalian cells. Adenoviruses encoding Saccharomyces cerevisiae HO endonuclease and its cleavage site were used to induce site-specific DSBs. Rolling circle amplification (RCA), a sensitive method that allows the detection of single molecular event by rapid isothermal amplification, was used to localize the broken ends in situ. Punctate RCA signals were only seen in the cells that had been infected with both adenoviruses encoding HO endonuclease and HO cleavage site, but not in the cells mock-infected or infected with the site or endonuclease virus only. With use of a chemical crosslinker, in situ RCA and immunofluorescence (IF) can be performed simultaneously on the same sample. This methodology provides a novel approach for investigation of DNA recombination, DNA repair, and checkpoint controls in mammalian cells.