Towards a formalism for mapping the spacetimes of massive compact objects: Bumpy black holes and their orbits

Observations have established that extremely compact, massive objects are common in the universe. It is generally accepted that these objects are black holes. As observations improve, it becomes possible to test this hypothesis in ever greater detail. In particular, it is or will be possible to measure the properties of orbits deep in the strong field of a black hole candidate (using x-ray timing or with gravitational-waves) and to test whether they have the characteristics of black hole orbits in general relativity. Such measurements can be used to map the spacetime of a massive compact object, testing whether the object's multipoles satisfy the strict constraints of the black hole hypothesis. Such a test requires that we compare against objects with the ``wrong'' multipole structure. In this paper, we present tools for constructing bumpy black holes: objects that are almost black holes, but that have some multipoles with the wrong value. The spacetimes which we present are good deep into the strong field of the object -- we do not use a large r expansion, except to make contact with weak field intuition. Also, our spacetimes reduce to the black hole spacetimes of general relativity when the ``bumpiness'' is set to zero. We propose bumpy black holes as the foundation for a null experiment: if black hole candidates are the black holes of general relativity, their bumpiness should be zero. By comparing orbits in a bumpy spacetime with those of an astrophysical source, observations should be able to test this hypothesis, stringently testing whether they are the black holes of general relativity. (Abridged)Comment: 16 pages + 2 appendices + 3 figures. Submitted to PR

Improved protocols for functional analysis in the pathogenic fungus Aspergillus flavus

<p>Abstract</p> <p>Background</p> <p>An available whole genome sequence for <it>Aspergillus flavus </it>provides the opportunity to characterize factors involved in pathogenicity and to elucidate the regulatory networks involved in aflatoxin biosynthesis. Functional analysis of genes within the genome is greatly facilitated by the ability to disrupt or mis-express target genes and then evaluate their result on the phenotype of the fungus. Large-scale functional analysis requires an efficient genetic transformation system and the ability to readily select transformants with altered expression, and usually requires generation of double (or multi) gene deletion strains or the use of prototrophic strains. However, dominant selectable markers, an efficient transformation system and an efficient screening system for transformants in <it>A. flavus </it>are absent.</p> <p>Results</p> <p>The efficiency of the genetic transformation system for <it>A. flavus </it>based on uracil auxotrophy was improved. In addition, <it>A. flavus </it>was shown to be sensitive to the antibiotic, phleomycin. Transformation of <it>A. flavus </it>with the <it>ble </it>gene for resistance to phleomycin resulted in stable transformants when selected on 100 μg/ml phleomycin. We also compared the phleomycin system with one based on complementation for uracil auxotrophy which was confirmed by uracil and 5-fluoroorotic acid selection and via transformation with the <it>pyr4 </it>gene from <it>Neurospora crassa </it>and <it>pyrG </it>gene from <it>A. nidulans </it>in <it>A. flavus </it>NRRL 3357. A transformation protocol using <it>pyr4 </it>as a selectable marker resulted in site specific disruption of a target gene. A rapid and convenient colony PCR method for screening genetically altered transformants was also developed in this study.</p> <p>Conclusion</p> <p>We employed phleomycin resistance as a new positive selectable marker for genetic transformation of <it>A. flavus</it>. The experiments outlined herein constitute the first report of the use of the antibiotic phleomycin for transformation of <it>A. flavus</it>. Further, we demonstrated that this transformation protocol could be used for directed gene disruption in <it>A. flavus</it>. The significance of this is twofold. First, it allows strains to be transformed without having to generate an auxotrophic mutation, which is time consuming and may result in undesirable mutations. Second, this protocol allows for double gene knockouts when used in conjunction with existing strains with auxotrophic mutations.</p> <p>To further facilitate functional analysis in this strain we developed a colony PCR-based method that is a rapid and convenient method for screening genetically altered transformants. This work will be of interest to those working on molecular biology of aflatoxin metabolism in <it>A. flavus</it>, especially for functional analysis using gene deletion and gene expression.</p

Cilia Have a Significant Role in Regulating Cell Size in Response to Fluid Flow Induced Shear Stress in a Flow Chamber

Cilia are hair-like protrusions on the apical surface of cells. Their function is to relay mechanical signals like shear stress from extracellular into intracellular environment and thereby maintain cellular homeostasis. Ciliary dysfunctions include polycystic kidney disease and new therapeutic interventions based on ciliary function are under investigation. The current study evaluates the use of a custom designed fluid flow chamber’s ability to study the role of cilia in regulating cell size in response to shear stress. A fluid flow chamber that continually maintains laminar flow at different flow rates and temperature was designed. Endothelial wild type cells (ETWT) that have cilia and polycystic kidney disease cells (PKD) that lost their ciliary function are grown on different glass slides. Cells on each glass slide are then exposed to continuous flow of phosphate-buffered saline at 37oC in the flow chamber. The optimal flow rate and duration of flow were first determined by measuring the total protein concentration before and after exposing the cells. Cell radius and area before and after exposing them to flow are measured using the NIS Software available on the microscope. The results from protein concentrations (n=12) indicate that cells are still attached at normal physiological flow rate 467 mL/min (2.8 µg/µL) and did not significantly differ from 60 mL/min (4.08 µg/µL) or 600 mL/min (2.73 µg/µL). The results for duration of fluid flow (n=22) show that 60 minutes (0.09 + 0.01 µg/µL) is optimal compared to 120 minutes (0.06 + 0.01 µg/µL) or 180 minutes (0.10 + 0.02 µg/µL). Under these optimal conditions, the average area of ETWT cells (n=300) measured from different slides before and after the flow is 4420.81+ 67.40 µm2 and 4678.17 + 87.15 µm2 (n=200) respectively. For PKD cells, the average area before and after the flow (n=300) is 5682.46 + 105.48 µm2 and 4173.74 + 263.97 µm2 (n=250). These results are in agreement with the published literature on the ability of cilia to maintain cell size in ETWT cells in response to shear stress that is similar to normal blood flow. However, under similar conditions, PKD cells could not maintain their cell size as the mechano-chemical signaling pathway that communicates external signals to prepare appropriate intracellular response is disrupted. These results provide confirmation that the custom designed parallel plate fluid flow chamber is a reliable tool to investigate the specific targets in the mechano-chemical cell signaling pathways

Balloon-borne coded aperture telescope for arc-minute angular resolution at hard x-ray energies

We are working on the development of a new balloon-borne telescope, MARGIE (minute-of-arc resolution gamma ray imaging experiment). It will be a coded aperture telescope designed to image hard x-rays (in various configurations) over the 20 - 600 keV range with an angular resolution approaching one arc minute. MARGIE will use one (or both) of two different detection plane technologies, each of which is capable of providing event locations with sub-mm accuracies. One such technology involves the use of cadmium zinc telluride (CZT) strip detectors. We have successfully completed a series of laboratory measurements using a prototype CZT detector with 375 micron pitch. Spatial location accuracies of better than 375 microns have been demonstrated. A second type of detection plane would be based on CsI microfiber arrays coupled to a large area silicon CCD readout array. This approach would provide spatial resolutions comparable to that of the CZT prototype. In one possible configuration, the coded mask would be 0.5 mm thick tungsten, with 0.5 mm pixels at a distance of 1.5 m from the central detector giving an angular resolution of 1 arc-minute and a fully coded field of view of 12 degrees. We review the capabilities of the MARGIE telescope and report on the status of our development efforts and our plans for a first balloon flight