Isolation of Primary Canine Satellite Cells

Duchenne muscular dystrophy (DMD) is a debilitating disease that principally affects striated muscles (skeletal and cardiac) and is the most severe form of muscular dystrophy. Disruption of the dystrophin gene is the primary cause of disease leading to excessive muscle damage. Regenerative processes counterbalance damage but individuals with DMD eventually succumb to immobilizing loss of strength and death from cardiac and pulmonary complications in their late teens and twenties. Golden retriever muscular dystrophy (GRMD) is a large animal model with better mimicry of the human disease than mouse models. Its development and characterization are critical to developing therapies for DMD. The cells primarily responsible for the regenerative response in skeletal muscle are satellite cells. These cells have been characterized at the protein level previously with only minor differences found between normal and dystrophic cultures. However, satellite cells have not been characterized at the transcriptional level. Pax7, MyoD, Myogenin and Utrophin act as critical members in the path to myogenesis. In this work, we have looked at the mRNA variation in cells collected from normal and GRMD animals and found substantial differences in mRNA expression profiles. These finding are also reflected in cell fusion experiments done on the same cultures. Studying these proteins and mRNAs in vitro under growth and differentiating conditions can help characterize satellite cells in the GRMD model. To sort through the heterogeneity of satellite cell populations, clonal cultures are needed to better characterize protein and mRNA patterns in these cells. Methods such as limiting dilution or flow cytometry require considerable time and resources to clone and verify large numbers of colonies for analysis. Micropallet array technology is a cell sorting method that permits clonal culture of large numbers of cells in very small spaces. Employing its flexible nature, micropallet array technology has been adapted to culture primary satellite cells from the GRMD model. Using these adaptations, clonal colonies have been cultured and shown to proliferate on tri-partite micropallet arrays. This forms two sister colonies where one sister colony can be analyzed and the other reserved for continued culture and downstream experiments

Lightweight XML-based query, integration and visualization of distributed, multimodality brain imaging data

A need of many neuroimaging researchers is to integrate multimodality brain data that may be stored in separate databases. To address this need we have developed a framework that provides a uniform XML-based query interface across multiple online data sources. The development of this framework is driven by the need to integrate neurosurgical and neuroimaging data related to language. The data sources for the language studies are 1) a web-accessible relational database of neurosurgical cortical stimulation mapping data (CSM) that includes patient-specific 3-D coordinates of each stimulation site mapped to an MRI reconstruction of the patient brain surface; and 2) an XML database of fMRI and structural MRI data and analysis results, created automatically by a batch program we have embedded in SPM. To make these sources available for querying each is wrapped as an XML view embedded in a web service. A top level web application accepts distributed XQueries over the sources, which are dispatched to the underlying web services. Returned results can be displayed as XML, HTML, CSV (Excel format), a 2-D schematic of a parcellated brain, or a 3-D brain visualization. In the latter case the CSM patient-specific coordinates returned by the query are sent to a transformation web-service for conversion to normalized space, after which they are sent to our 3-D visualization program MindSeer, which is accessed via Java WebStart through a generated link. The anatomical distribution of pooled CSM sites can then be visualized using various surfaces derived from brain atlases. As this framework is further developed and generalized we believe it will have appeal for researchers who wish to query, integrate and visualize results across their own databases as well as those of collaborators

Distributed XQuery-based integration and visualization of multimodality data: Application to brain mapping.

This paper addresses the need for relatively small groups of collaborating investigators to integrate distributed and heterogeneous data about the brain. Although various national efforts facilitate large-scale data sharing, these approaches are generally too “heavyweight” for individual or small groups of investigators, with the result that most data sharing among collaborators continues to be ad hoc. Our approach to this problem is to create a “lightweight” distributed query architecture, in which data sources are accessible via web services that accept arbitrary query languages but return XML results. A Distributed XQuery Processor (DXQP) accepts distributed XQueries in which subqueries are shipped to the remote data sources to be executed, with the resulting XML integrated by DXQP. A web-based application called DXBrain accesses DXQP, allowing a user to create, save and execute distributed XQueries, and to view the results in various formats including a 3-D brain visualization. Example results are presented using distributed brain mapping data sources obtained in studies of language organization in the brain, but any other XML source could be included. The advantage of this approach is that it is very easy to add and query a new source, the tradeoff being that the user needs to understand XQuery and the schemata of the underlying sources. For small numbers of known sources this burden is not onerous for a knowledgeable user, leading to the conclusion that the system helps to fill the gap between ad hoc local methods and large scale but complex national data sharing efforts

Polystyrene-coated micropallets for culture and separation of primary muscle cells

Despite identification of a large number of adult stem cell types, current primary cell isolation and identification techniques yield heterogeneous samples, making detailed biological studies challenging. To identify subsets of isolated cells, technologies capable of simultaneous cell culture and cloning are necessary. Micropallet arrays, a new cloning platform for adherent cell types, hold great potential. However, the microstructures composing these arrays are fabricated from an epoxy photoresist 1002F, a growth surface unsuitable for many cell types. Optimization of the microstructures’ surface properties was conducted for the culture of satellite cells, primary muscle cells for which improved cell isolation techniques are desired. A variety of surface materials were screened for satellite cell adhesion and proliferation and compared to their optimal substrate, gelatin-coated Petri dishes. A 1-μm thick, polystyrene copolymer was applied to the microstructures by contact-printing. A negatively charged copolymer of 5% acrylic acid in 95% styrene was found to be equivalent to the control Petri dishes for cell adhesion and proliferation. Cells cultured on control dishes and optimal copolymer-coated surfaces maintained an undifferentiated state and showed similar mRNA expression for two genes indicative of cell differentiation during a standard differentiation protocol. Experiments using additional contact-printed layers of extracellular matrix proteins collagen and gelatin showed no further improvements. This micropallet coating strategy is readily adaptable to optimize the array surface for other types of primary cells

Investigation of \u3csup\u3e186\u3c/sup\u3eRe via radiative thermal-neutron capture on \u3csup\u3e185\u3c/sup\u3eRe

Partial -ray production cross sections and the total radiative thermal-neutron capture cross section for the 185Re(n,)186Re reaction were measured using the Prompt Gamma Activation Analysis facility at the Budapest Research Reactor with an enriched 185Re target. The 186Re cross sections were standardized using well-known 35Cl(n,)36Cl cross sections from irradiation of a stoichiometric natReCl3 target. The resulting cross sections for transitions feeding the 186Re ground state from low-lying levels below a cutoff energy of Ec=746keV were combined with a modeled probability of ground-state feeding from levels above Ec to arrive at a total cross section of σ0=111(6)b for radiative thermal-neutron capture on 185Re. A comparison of modeled discrete-level populations with measured transition intensities led to proposed revisions for seven tentative spin-parity assignments in the adopted level scheme for 186Re. Additionally, 102 primary rays were measured, including 50 previously unknown. A neutron-separation energy of Sn=6179.59(5)keV was determined from a global least-squares fit of the measured -ray energies to the known 186Re decay scheme. The total capture cross section and separation energy results are comparable to earlier measurements of these values

Schottky mass measurements of heavy neutron-rich nuclides in the element range 70\leZ \le79 at the ESR

Storage-ring mass spectrometry was applied to neutron-rich $^{197}$Au projectile fragments. Masses of $^{181,183}$Lu, $^{185,186}$Hf, $^{187,188}$Ta, $^{191}$W, and $^{192,193}$Re nuclei were measured for the first time. The uncertainty of previously known masses of $^{189,190}$W and $^{195}$Os nuclei was improved. Observed irregularities on the smooth two-neutron separation energies for Hf and W isotopes are linked to the collectivity phenomena in the corresponding nuclei.Comment: 10 pages, 9 figures, 2 table

Deep Underground Science and Engineering Laboratory - Preliminary Design Report

The DUSEL Project has produced the Preliminary Design of the Deep Underground Science and Engineering Laboratory (DUSEL) at the rehabilitated former Homestake mine in South Dakota. The Facility design calls for, on the surface, two new buildings - one a visitor and education center, the other an experiment assembly hall - and multiple repurposed existing buildings. To support underground research activities, the design includes two laboratory modules and additional spaces at a level 4,850 feet underground for physics, biology, engineering, and Earth science experiments. On the same level, the design includes a Department of Energy-shepherded Large Cavity supporting the Long Baseline Neutrino Experiment. At the 7,400-feet level, the design incorporates one laboratory module and additional spaces for physics and Earth science efforts. With input from some 25 science and engineering collaborations, the Project has designed critical experimental space and infrastructure needs, including space for a suite of multidisciplinary experiments in a laboratory whose projected life span is at least 30 years. From these experiments, a critical suite of experiments is outlined, whose construction will be funded along with the facility. The Facility design permits expansion and evolution, as may be driven by future science requirements, and enables participation by other agencies. The design leverages South Dakota's substantial investment in facility infrastructure, risk retirement, and operation of its Sanford Laboratory at Homestake. The Project is planning education and outreach programs, and has initiated efforts to establish regional partnerships with underserved populations - regional American Indian and rural populations

Developments in Capture- γ Libraries for Nonproliferation Applications

The neutron-capture reaction is fundamental for identifying and analyzing the γ-ray spectrum from an unknown assembly because it provides unambiguous information on the neutron-absorbing isotopes. Nondestructive-assay applications may exploit this phenomenon passively, for example, in the presence of spontaneous-fission neutrons, or actively where an external neutron source is used as a probe. There are known gaps in the Evaluated Nuclear Data File libraries corresponding to neutron-capture γ-ray data that otherwise limit transport-modeling applications. In this work, we describe how new thermal neutron-capture data are being used to improve information in the neutron-data libraries for isotopes relevant to nonproliferation applications. We address this problem by providing new experimentally-deduced partial and total neutron-capture reaction cross sections and then evaluate these data by comparison with statistical-model calculations

\textsc{MaGe} - a {\sc Geant4}-based Monte Carlo Application Framework for Low-background Germanium Experiments

We describe a physics simulation software framework, MAGE, that is based on the GEANT4 simulation toolkit. MAGE is used to simulate the response of ultra-low radioactive background radiation detectors to ionizing radiation, specifically the MAJORANA and GERDA neutrinoless double-beta decay experiments. MAJORANA and GERDA use high-purity germanium detectors to search for the neutrinoless double-beta decay of 76Ge, and MAGE is jointly developed between these two collaborations. The MAGE framework contains the geometry models of common objects, prototypes, test stands, and the actual experiments. It also implements customized event generators, GEANT4 physics lists, and output formats. All of these features are available as class libraries that are typically compiled into a single executable. The user selects the particular experimental setup implementation at run-time via macros. The combination of all these common classes into one framework reduces duplication of efforts, eases comparison between simulated data and experiment, and simplifies the addition of new detectors to be simulated. This paper focuses on the software framework, custom event generators, and physics lists.Comment: 12 pages, 6 figure