Municipal Torts: The Rule Is Liability—The Exception Is Immunity

Enghauser Manufacturing Co. v. Eriksson Engineering Ltd., 6 Ohio St. 3d 31, 451 N.E.2d 228 (1983)

Evaluation of the table Mountain Ronchi telescope for angular tracking

The performance of the University of California at San Diego (UCSD) Table Mountain telescope was evaluated to determine the potential of such an instrument for optical angular tracking. This telescope uses a Ronchi ruling to measure differential positions of stars at the meridian. The Ronchi technique is summarized and the operational features of the Table Mountain instrument are described. Results from an analytic model, simulations, and actual data are presented that characterize the telescope's current performance. For a star pair of visual magnitude 7, the differential uncertainty of a 5-min observation is about 50 nrad (10 marcsec), and tropospheric fluctuations are the dominant error source. At magnitude 11, the current differential uncertainty is approximately 800 nrad (approximately 170 marcsec). This magnitude is equivalent to that of a 2-W laser with a 0.4-m aperture transmitting to Earth from a spacecraft at Saturn. Photoelectron noise is the dominant error source for stars of visual magnitude 8.5 and fainter. If the photoelectron noise is reduced, ultimately tropospheric fluctuations will be the limiting source of error at an average level of 35 nrad (7 marcsec) for stars approximately 0.25 deg apart. Three near-term strategies are proposed for improving the performance of the telescope to the 10-nrad level: improving the efficiency of the optics, masking background starlight, and averaging tropospheric fluctuations over multiple observations

The Celestial Reference Frame at 24 and 43 GHz. II. Imaging

We have measured the sub-milli-arcsecond structure of 274 extragalactic sources at 24 and 43 GHz in order to assess their astrometric suitability for use in a high frequency celestial reference frame (CRF). Ten sessions of observations with the Very Long Baseline Array have been conducted over the course of $\sim$5 years, with a total of 1339 images produced for the 274 sources. There are several quantities that can be used to characterize the impact of intrinsic source structure on astrometric observations including the source flux density, the flux density variability, the source structure index, the source compactness, and the compactness variability. A detailed analysis of these imaging quantities shows that (1) our selection of compact sources from 8.4 GHz catalogs yielded sources with flux densities, averaged over the sessions in which each source was observed, of about 1 Jy at both 24 and 43 GHz, (2) on average the source flux densities at 24 GHz varied by 20%-25% relative to their mean values, with variations in the session-to-session flux density scale being less than 10%, (3) sources were found to be more compact with less intrinsic structure at higher frequencies, and (4) variations of the core radio emission relative to the total flux density of the source are less than 8% on average at 24 GHz. We conclude that the reduction in the effects due to source structure gained by observing at higher frequencies will result in an improved CRF and a pool of high-quality fiducial reference points for use in spacecraft navigation over the next decade.Comment: 63 pages, 18 figures, 6 tables, accepted by the Astronomical Journa

Co-expression and purification of the RadA recombinase with the RadB paralog from Haloferax volcanii yields heteromeric ring-like structures

The study of archaeal proteins and the processes to which they contribute poses particular challenges due to the often extreme environments in which they function. DNA recombination, replication and repair proteins of the halophilic euryarchaeon, Haloferax volcanii (Hvo) are of particular interest as they tend to resemble eukaryotic counterparts in both structure and activity, and genetic tools are available to facilitate their analysis. In the present study, we show using bioinformatics approaches that the Hvo RecA-like protein RadA is structurally similar to other recombinases although is distinguished by a unique acidic insertion loop. To facilitate expression of Hvo RadA a co-expression approach was used, providing its lone paralog, RadB, as a binding partner. At present, structural and biochemical characterization of Hvo RadA is lacking. Here, we describe for the first time co-expression of Hvo RadA with RadB and purification of these proteins as a complex under in vitro conditions. Purification procedures were performed under high salt concentration (>1 M sodium chloride) to maintain the solubility of the proteins. Quantitative densitometry analysis of the co-expressed and co-purified RadAB complex estimated the ratio of RadA to RadB to be 4 : 1, which suggests that the proteins interact with a specific stoichiometry. Based on a combination of analyses, including size exclusion chromatography, Western blot and electron microscopy observations, we suggest that RadA multimerizes into a ring-like structure in the absence of DNA and nucleoside co-factor

Protein Conformational Changes in the Bacteriorhodopsin Photocycle: Comparison of Findings from Electron and X-Ray Crystallographic Analyses

Light-driven conformational changes in the membrane protein bacteriorhodopsin have been studied extensively using X-ray and electron crystallography, resulting in the deposition of >30 sets of coordinates describing structural changes at various stages of proton transport. Using projection difference Fourier maps, we show that coordinates reported by different groups for the same photocycle intermediates vary considerably in the extent and nature of conformational changes. The different structures reported for the same intermediate cannot be reconciled in terms of differing extents of change on a single conformational trajectory. New measurements of image phases obtained by cryo-electron microscopy of the D96G/F171C/F219L triple mutant provide independent validation for the description of the large protein conformational change derived at 3.2 Å resolution by electron crystallography of 2D crystals, but do not support atomic models for light-driven conformational changes derived using X-ray crystallography of 3D crystals. Our findings suggest that independent determination of phase information from 2D crystals can be an important tool for testing the accuracy of atomic models for membrane protein conformational changes

Relocation of Advanced Water Vapor Radiometer 1 to Deep Space Station 55

In June of 2004, the Advanced Water Vapor Radiometer (AWVR) unit no. 1 was relocated to the Deep Space Station (DSS) 55 site in Madrid, Spain, from DSS 25 in Goldstone, California. This article summarizes the relocation activity and the subsequent operation and data acquisition. This activity also relocated the associated Microwave Temperature Profiler (MTP) and Surface Meteorology (SurfMET) package that collectively comprise the Cassini Media Calibration System (MCS)

Rotational Alignment Altered by Source Position Correlations

In the construction of modern Celestial Reference Frames (CRFs) the overall rotational alignment is only weakly constrained by the data. Therefore, common practice has been to apply a 3-dimensional No-Net-Rotation (NNR) constraint in order to align an under-construction frame to the ICRF. We present evidence that correlations amongst source position parameters must be accounted for in order to properly align a CRF at the 5-10 (mu)as level of uncertainty found in current work. Failure to do so creates errors at the 10-40 (mu)as level

Single-Molecule FRET TACKLE Reveals Highly Dynamic Mismatched DNA−MutS Complexes

The first step in DNA mismatch repair (MMR) is the recognition of DNA mismatches or nucleotide insertions/deletions (IDLs) by MutS and MutS homologs. To investigate the conformational properties of MutS-mismatch complexes, we used single-molecule fluorescence resonance energy transfer (smFRET) to examine the dynamics of MutS-induced DNA bending at a GT-mismatch. The FRET measurements reveal that the MutS-GT-mismatch recognition complex is highly dynamic, undergoing conformational transitions between many states with different degrees of DNA bending. Due to the complexity of the data, we developed an analysis approach, called FRET TACKLE, in which we combine direct analysis of FRET transitions with examination of kinetic lifetimes to identify all of the conformational states and characterize the kinetics of the binding and conformational equilibria. The data reveal that MutS-GT complexes can reside in six different conformations, which have lifetimes that differ by as much as 20-fold and exhibit rates of interconversion that vary by two orders of magnitude. To gain further insight into the dynamic properties of GT-MutS complexes and to bolster the validity of our analysis, we complemented our experimental data with Monte Carlo simulations. Taken together, our results suggest that the dynamics of the MutS-mismatch complex could govern the efficiency of repair of different DNA mismatches. Finally, in addition to revealing these important biological implications of MutS-DNA interactions, this FRET TACKLE method will enable the analysis of the complex dynamics of other biological systems

Identification of acetyltransferase genes (HAT1 and KAT8) regulating HBV replication by RNAi screening

Background: The initiation of hepatitis B virus (HBV) replication involves the formation of covalently closed circular DNA (cccDNA) and its transcription into pregenomic RNA (pgRNA) in hepatocyte nuclei. The regulatory mechanism of HBV replication by acetyltransferase is thus far not well understood, but human acetyltransferase has been reported as being involved in the regulation of HBV replication. Results: Depletion of KAT8 or HAT1 via RNA interference (RNAi) markedly down-regulated HBV-DNA and pgRNA levels in HepG2.2.15 cells, with KAT8 knockdown reducing both HBsAg and HBeAg more than HAT1 knockdown. Con-sistent with these observations, HBV replication regulators hepatocyte nuclear factor-4-α (HNF4α) and peroxisome proliferator-activated receptor gamma coactivator- (PPARGC-) 1-α were decreased following knockdown of HAT1 or KAT8. Conclusions: These data suggest that KAT8 or HAT1 regulate HBV replication and may be potential drug targets of anti-HBV therapy

DSN Beowulf Cluster-Based VLBI Correlator

The NASA Deep Space Network (DSN) requires a broadband VLBI (very long baseline interferometry) correlator to process data routinely taken as part of the VLBI source Catalogue Maintenance and Enhancement task (CAT M&E) and the Time and Earth Motion Precision Observations task (TEMPO). The data provided by these measurements are a crucial ingredient in the formation of precision deep-space navigation models. In addition, a VLBI correlator is needed to provide support for other VLBI related activities for both internal and external customers. The JPL VLBI Correlator (JVC) was designed, developed, and delivered to the DSN as a successor to the legacy Block II Correlator. The JVC is a full-capability VLBI correlator that uses software processes running on multiple computers to cross-correlate two-antenna broadband noise data. Components of this new system (see Figure 1) consist of Linux PCs integrated into a Beowulf Cluster, an existing Mark5 data storage system, a RAID array, an existing software correlator package (SoftC) originally developed for Delta DOR Navigation processing, and various custom- developed software processes and scripts. Parallel processing on the JVC is achieved by assigning slave nodes of the Beowulf cluster to process separate scans in parallel until all scans have been processed. Due to the single stream sequential playback of the Mark5 data, some ramp-up time is required before all nodes can have access to required scan data. Core functions of each processing step are accomplished using optimized C programs. The coordination and execution of these programs across the cluster is accomplished using Pearl scripts, PostgreSQL commands, and a handful of miscellaneous system utilities. Mark5 data modules are loaded on Mark5 Data systems playback units, one per station. Data processing is started when the operator scans the Mark5 systems and runs a script that reads various configuration files and then creates an experiment-dependent status database used to delegate parallel tasks between nodes and storage areas (see Figure 2). This script forks into three processes: extract, translate, and correlate. Each of these processes iterates on available scan data and updates the status database as the work for each scan is completed. The extract process coordinates and monitors the transfer of data from each of the Mark5s to the Beowulf RAID storage systems. The translate process monitors and executes the data conversion processes on available scan files, and writes the translated files to the slave nodes. The correlate process monitors the execution of SoftC correlation processes on the slave nodes for scans that have completed translation. A comparison of the JVC and the legacy Block II correlator outputs reveals they are well within a formal error, and that the data are comparable with respect to their use in flight navigation. The processing speed of the JVC is improved over the Block II correlator by a factor of 4, largely due to the elimination of the reel-to-reel tape drives used in the Block II correlator