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At least 217 records · Page 12

Quantitative method of measuring cancer cell urokinase and metastatic potential

The metastatic potential of tumors can be evaluated by the quantitative detection of urokinase and DNA. The cell sample selected for examination is analyzed for the presence of high levels of urokinase and abnormal DNA using analytical flow cytometry and digital image analysis. Other factors such as membrane associated urokinase, increased DNA synthesis rates and certain receptors can be used in the method for detection of potentially invasive tumors.

Morrison, Dennis R.↗

Quantitative Method of Measuring Metastatic Activity

The metastatic potential of tumors can be evaluated by the quantitative detection of urokinase and DNA. The cell sample selected for examination is analyzed for the presence of high levels of urokinase and abnormal DNA using analytical flow cytometry and digital image analysis. Other factors such as membrane associated uroldnase, increased DNA synthesis rates and certain receptors can be used in the method for detection of potentially invasive tumors.

Morrison, Dennis R.↗

Laser microsurgery of higher plant cell walls permits patch-clamp access

Plasma membranes of guard cells in epidermal peels of Vicia faba and Commelina communis can be made accessible to a patch-clamp pipet by removing a small portion (1-3 micrometers in diameter) of the guard cell wall using a microbeam of ultraviolet light generated by a nitrogen laser. Using this laser microsurgical technique, we have measured channel activity across plasma membranes of V. faba guard cells in both cell-attached and isolated patch configurations. Measurements made in the inside-out patch configuration revealed two distinct K(+)-selective channels. Major advantages of the laser microsurgical technique include the avoidance of enzymatic protoplast isolation, the ability to study cell types that have been difficult to isolate as protoplasts or for which enzymatic isolation protocols result in protoplasts not amenable to patch-clamp studies, the maintenance of positional information in single-channel measurements, reduced disruption of cell-wall-mediated signaling pathways, and the ability to investigate intercellular signaling through studies of cells remaining situated within tissue.

Non-NASA Center↗

Protonic Conduction of BaCe0.85YO. 1503 Doped with SrTiO3

Reformers based on ceramic membrane technology potentially offer hydrogen production that is comparable to the cost of fossil fuels. Protonic conducting ceramic with the chemical formula AB03 offers the promise of highly selective hydrogen separation at intermediate temperature (400-800 C). Among different perovskite-type oxides, BaCe03 and SrCe03 based compositions show high protonic conductivities but strong resistance to densification. X-ray diffraction studies on sintered specimens of BaCe0.85Y0.1503-6 show multi-phase formation which was found to show dependence upon powder synthesis method. Doping with SrTiO3 suppresses multi-phase formation and enhances grain growth. Conductivity measurements in temperature range of 200 to 1000 C were performed by ac impedance spectroscopy under dry and wet conditions. Sintering behavior, phase formation and conductivity results will be reported.

Dynys, Frederick W.↗

Benchtop Detection of Proteins

A process, and a benchtop-scale apparatus for implementing the process, have been developed to detect proteins associated with specific microbes in water. The process and apparatus may also be useful for detection of proteins in other, more complex liquids. There may be numerous potential applications, including monitoring lakes and streams for contamination, testing of blood and other bodily fluids in medical laboratories, and testing for microbial contamination of liquids in restaurants and industrial food-processing facilities. A sample can be prepared and analyzed by use of this process and apparatus within minutes, whereas an equivalent analysis performed by use of other processes and equipment can often take hours to days. The process begins with the conjugation of near-infrared-fluorescent dyes to antibodies that are specific to a particular protein. Initially, the research has focused on using near-infrared dyes to detect antigens or associated proteins in solution, which has proven successful vs. microbial cells, and streamlining the technique in use for surface protein detection on microbes would theoretically render similar results. However, it is noted that additional work is needed to transition protein-based techniques to microbial cell detection. Consequently, multiple such dye/antibody pairs could be prepared to enable detection of multiple selected microbial species, using a different dye for each species. When excited by near-infrared light of a suitable wavelength, each dye fluoresces at a unique longer wavelength that differs from those of the other dyes, enabling discrimination among the various species. In initial tests, the dye/antibody pairs are mixed into a solution suspected of containing the selected proteins, causing the binding of the dye/antibody pairs to such suspect proteins that may be present. The solution is then run through a microcentrifuge that includes a membrane that acts as a filter in that it retains the dye/antibody/protein complexes while allowing any remaining unbound dye/antibody pairs to flow away. The retained dye/antibody/protein complexes are transferred to a cuvette, wherein they are irradiated with light from a miniature near-infrared laser delivered via a fiber-optic cable. The resulting fluorescence from the dye(s) is measured by use of a miniature spectrometer, the output of which is digitized, then analyzed by laptop computer. The software running in the computer identifies the protein species by the wavelengths of their spectral peaks and determines the amounts of the proteins, and thus, one day, microbes of the various species from the intensities of the peaks. The abovementioned removal of the unbound dye/antibody pairs during centrifugation prevents false positive readings. The process proves successful in detecting proteins in solution and thus can now be employed for use in microbe detection.

Scardelletti, Maximilian C.↗

Analytical displacements and vibrations of cantilevered unsymmetric fiber composite laminates

A fiber composite flat cantilever plate that has symmetric and nonsymmetric laminate configurations is theoretically investigated to determine its static and dynamic structural response. The finite element analysis method used includes a unique triangular finite element developed at Lewis for the analysis of fiber composite airfoils. The various responses investigated include tip displacements, natural frequencies, and fundamental mode shapes. The results show that the displacements and the natural frequencies can be in considerable error for nonsymmetric laminate configurations if the membrane-bending coupling is not taken into account. Structural response results obtained from ten laminate configurations are presented in tabular forms and may be used as an aid in selecting laminate configurations for composite airfoils.

Minich, M. D.↗

Bucky Paper as a Support Membrane in Retinal Cell Transplantation

A method for repairing a retinal system of an eye, using bucky paper on which a plurality of retina pigment epithelial cells and/or iris pigment epithelial cells and/or stem cells is deposited, either randomly or in a selected cell pattern. The cell-covered bucky paper is positioned in a sub-retinal space to transfer cells to this space and thereby restore the retina to its normal functioning, where retinal damage or degeneration, such as macular degeneration, has occurred.

Loftus, David J.↗

Centrifugal Adsorption Cartridge System

The centrifugal adsorption cartridge system (CACS) is an apparatus that recovers one or more bioproduct(s) from a dilute aqueous solution or suspension flowing from a bioreactor. The CACS can be used both on Earth in unit gravity and in space in low gravity. The CACS can be connected downstream from the bioreactor; alternatively, it can be connected into a flow loop that includes the bioreactor so that the liquid can be recycled. A centrifugal adsorption cartridge in the CACS (see figure) includes two concentric cylinders with a spiral ramp between them. The volume between the inner and outer cylinders, and between the turns of the spiral ramp is packed with an adsorbent material. The inner cylinder is a sieve tube covered with a gas-permeable, hydrophobic membrane. During operation, the liquid effluent from the bioreactor is introduced at one end of the spiral ramp, which then constrains the liquid to flow along the spiral path through the adsorbent material. The spiral ramp also makes the flow more nearly uniform than it would otherwise be, and it minimizes any channeling other than that of the spiral flow itself. The adsorbent material is formulated to selectively capture the bioproduct(s) of interest. The bioproduct(s) can then be stored in bound form in the cartridge or else eluted from the cartridge. The centrifugal effect of the spiral flow is utilized to remove gas bubbles from the liquid. The centrifugal effect forces the bubbles radially inward, toward and through the membrane of the inner cylinder. The gas-permeable, hydrophobic membrane allows the bubbles to enter the inner cylinder while keeping the liquid out. The bubbles that thus enter the cylinder are vented to the atmosphere. The spacing between the ramps determines rate of flow along the spiral, and thereby affects the air-bubble-removal efficiency. The spacing between the ramps also determines the length of the fluid path through the cartridge adsorbent, and thus affects the bioproduct-capture efficiency of the cartridge. Depending on the application, several cartridges could be connected in a serial or parallel flow arrangement. A parallel arrangement can be used to increase product-capturing and flow capacities while maintaining a low pressure drop. A serial arrangement can be used to obtain high product-capturing capacity; alternatively, series-connected cartridges can be packed with different adsorbents to capture different bioproducts simultaneously.

Gonda, Steve R.↗

Peptides at Membrane Surfaces and their Role in the Origin of Life

All ancestors of contemporary cells (protocells) had to transport ions and organic matter across membranous walls, capture and utilize energy and transduce environmental signals. In modern organisms, all these functions are preformed by membrane proteins. We make the parsimonious assumption that in the protobiological milieu the same functions were carried out by their simple analogs - peptides. This, however, required that simple peptides could self-organize into ordered, functional structures. In a series of detailed, molecular-level computer simulations we demonstrated how this is possible. One example is the peptide (LSLLLSL)3 which forms a trameric bundle capable of transporting protons across membranes. Another example is the transmembrane pore of the influenza M2 protein. This aggregate of four identical alpha-helices, each built of 25 amino acids, forms an efficient and selective voltage-gated proton channel. Our simulations explain the gating mechanism in this channel. The channel can be re-engineered into a simple proton pump.

Pohorille, Andrew↗

Triamcinolone Acetonide Selectively Inhibits Angiogenesis in Small Blood Vessels and Decreases Vessel Diameter within the Vascular Tree

The steroid triamcinolone acetonide (TA) is a potent anti-angiogenesis drug used to treat retinal vascular diseases that include diabetic retinopathy, vascular occlusions and choroidal neovascularization. To quantify the effects of TA on branching morphology within the angiogenic microvascular tree of the chorioallantoic membrane (CAM) of quail embryos. Increasing concentrations of TA (0-16 ng/ml) were applied topically on embryonic day 7 (E7) to the chorioallantoic membrane (CAM) of quail embryos cultured in Petri dishes, and incubated for an additional 24 or 48 hours until fixation. Binary (black/white) microscopic images of arterial end points were quantified by VESGEN software (for Generational Analysis of Vessel Branching) to obtain major vascular parameters that include vessel diameter (Dv), fractal dimension (Df), tortuosity (Tv) and densities of vessel area, length, number and branch point (Av, Lv, Nv and Brv). For assessment of specific changes in vascular morphology induced by TA, the VESGEN software automatically segmented the vascular tree into branching generations (G1...G10) according to changes in vessel diameter and branching. Vessel density decreased significantly up to 34% as the function of increasing concentration of TA according to Av, Lv, Brv, Nv and Df. TA selectively inhibited the growth of new, small vessels, because Lv decreased from 13.14plus or minus 0.61 cm/cm2 for controls to 8.012 plus or minus 0.82 cm/cm2 at 16 ng TA/ml in smaller branching generations (G7-G10), and for Nv from 473.83 plus or minus 29.85 cm(-)2 to 302.32 plus or minus 33.09 cm-()2. In contrast, vessel diameter (Dv) decreased throughout the vascular tree (G1-G10).

McKay, Terri L.↗

Elastic Distribution of Microshutters, Measurements Obtainable on James Web Space Telescope

Spectrographic astronomy measurements in the near-infrared region will be done by functional two-dimensional microshutter arrays that are being fabricated at the NASA Goddard Space Flight Center for the James Webb Space Telescope (JWST). These micro-shutter arrays will represent the first mission-critical MEMS devices to be flown in space. JWST will use microshutter arrays to select focal plane object. 2-D programmable aperture masks of more than 200,000 elements select such space object. The use of silicon wafer material promises high efficiency and high contrast. Microshutter operation temperature is around 35K. Microshutter arrays are fabricated as close-packed silicon nitride membranes with a unit cell size of 105 x 204 micrometers. A layer of magnetic material is deposited onto each shutter. Individual shutters are equipped with a torsion flexure. Reactive ion etching (RIE) releases the shutters so they can open up to 90 degrees using the torsion flexure. Shutter rotation is initiated into a silicon support structure via an external magnetic field. Two electrically independent aluminum electrodes are deposited, one onto each shutter and another onto the support structure side-wall, permitting electrostatic latching and 2-D addressing to hold specific shutters open via external electronics.

Kletetschka, Gunther↗

Buckling Of Long Compression-Loaded Anisotropic Plates Restrained Against Inplane Lateral and Shear Deformations

An approach for synthesizing buckling results and behavior for thin balanced and unbalanced symmetric laminates that are subjected to uniform axial compression loads and elastically restrained against inplane expansion, contraction, and shear deformation is presented. This approach uses a nondimensional analysis for infinitely long, flexurally anisotropic plates (coupling between bending and twisting) that are subjected to combined mechanical loads and is based on nondimensional parameters. In addition, nondimensional loading parameters are derived that account for the effects of the elastic inplane deformation restraints, membrane orthotropy, and membrane anisotropy on the induced prebuckling stress state. The loading parameters are used to determine buckling coefficients that include the effects of flexural orthotropy and flexural anisotropy. Many results are presented, for some selected laminates, that are intended to facilitate a structural designer's transition to the use of the generic buckling design curves that are presented and discussed in the paper. Several buckling response curves are presented that provide physical insight into the behavior for combined loads, in addition to providing useful design data. An example is presented that demonstrates the use of the generic design curves, which are applicable to a wide range of laminate constructions. The analysis approach and generic results indicate the effects and characteristics of laminate orthotropy and anisotropy in a very general and unifying manner.

Nemeth, Michael P.↗

Multi-Functional Sandwich Composites for Spacecraft Applications: An Initial Assessment

Current spacecraft implement relatively uncoupled material and structural systems to address a variety of design requirements, including structural integrity, damage tolerance, radiation protection, debris shielding and thermal insulation. This investigation provided an initial assessment of multi-functional sandwich composites to integrate these diverse requirements. The need for radiation shielding was addressed through the selection of polymeric constituents with high hydrogen content. To provide increased damage tolerance and debris shielding, manufacturing techniques were developed to incorporate transverse stitching reinforcement, internal layers, and a self-healing ionomer membrane. To assess the effects of a space environment, thermal expansion behavior of the candidate foam materials was investigated under a vacuum and increasing temperature. Finally, a thermal expansion model was developed for foam under vacuum conditions and its predictive capability assessed.

Adams, Daniel O.↗

PMA-PhyloChip DNA Microarray to Elucidate Viable Microbial Community Structure

Since the Viking missions in the mid-1970s, traditional culture-based methods have been used for microbial enumeration by various NASA programs. Viable microbes are of particular concern for spacecraft cleanliness, for forward contamination of extraterrestrial bodies (proliferation of microbes), and for crew health/safety (viable pathogenic microbes). However, a "true" estimation of viable microbial population and differentiation from their dead cells using the most sensitive molecular methods is a challenge, because of the stability of DNA from dead cells. The goal of this research is to evaluate a rapid and sensitive microbial detection concept that will selectively estimate viable microbes. Nucleic acid amplification approaches such as the polymerase chain reaction (PCR) have shown promise for reducing time to detection for a wide range of applications. The proposed method is based on the use of a fluorescent DNA intercalating agent, propidium monoazide (PMA), which can only penetrate the membrane of dead cells. The PMA-quenched reaction mixtures can be screened, where only the DNA from live cells will be available for subsequent PCR reaction and microarray detection, and be identified as part of the viable microbial community. An additional advantage of the proposed rapid method is that it will detect viable microbes and differentiate from dead cells in only a few hours, as opposed to less comprehensive culture-based assays, which take days to complete. This novel combination approach is called the PMA-Microarray method. DNA intercalating agents such as PMA have previously been used to selectively distinguish between viable and dead bacterial cells. Once in the cell, the dye intercalates with the DNA and, upon photolysis under visible light, produces stable DNA adducts. DNA cross-linked in this way is unavailable for PCR. Environmental samples suspected of containing a mixture of live and dead microbial cells/spores will be treated with PMA, and then incubated in the dark. Thereafter, the sample is exposed to visible light for five minutes, so that the DNA from dead cells will be cross-linked. Following this PMA treatment step, the sample is concentrated by centrifugation and washed (to remove excessive PMA) before DNA is extracted. The 16S rRNA gene fragments will be amplified by PCR to screen the total microbial community using PhyloChip DNA microarray analysis. This approach will detect only the viable microbial community since the PMA intercalated DNA from dead cells would be unavailable for PCR amplification. The total detection time including PCR reaction for low biomass samples will be a few hours. Numerous markets may use this technology. The food industry uses spore detection to validate new alternative food processing technologies, sterility, and quality. Pharmaceutical and medical equipment companies also detect spores as a marker for sterility. This system can be used for validating sterilization processes, water treatment systems, and in various public health and homeland security applications.

Venkateswaran, Kasthuri J.↗

Spacesuit Water Membrane Evaporator Development for Lunar Missions

For future lunar extra-vehicular activities (EVA), one method under consideration for rejecting crew and electronics heat involves evaporating water through a hydrophobic, porous Teflon membrane. A Spacesuit Water Membrane Evaporator (SWME) prototype using the Teflon membrane was tested successfully by Ungar and Thomas (2001) with predicted performance matching test data well. The above referenced work laid the foundation for the design of the SWME development unit, which is being considered for service in the Constellation System Spacesuit Element (CSSE) Portable Life Support System (PLSS). Multiple PLSS SWME configurations were considered on the basis of thermal performance, mass, volume, and performance and manufacturing risk. All configurations were a variation of an alternating concentric water and vapor channel configuration or a stack of alternating rectangular water and vapor channels. Supporting thermal performance trades mapped maximum SWME heat rejection as a function of water channel thickness, vapor channel thickness, channel length, number of water channels, porosity of the membrane structural support, and backpressure valve throat area. Preliminary designs of each configuration were developed to determine total mass and volume as well as to understand manufacturing issues. Review of configurations led to the selection of a concentric annulus configuration that meets the requirements of 800 watts (W) of heat rejection. Detailed design of the SWME development unit will be followed by fabrication of a prototype test unit, with thermal testing expected to start in 2008.

Vogel, Matt R.↗

Tissue Photolithography

Tissue lithography will enable physicians and researchers to obtain macromolecules with high purity (greater than 90 percent) from desired cells in conventionally processed, clinical tissues by simply annotating the desired cells on a computer screen. After identifying the desired cells, a suitable lithography mask will be generated to protect the contents of the desired cells while allowing destruction of all undesired cells by irradiation with ultraviolet light. The DNA from the protected cells can be used in a number of downstream applications including DNA sequencing. The purity (i.e., macromolecules isolated form specific cell types) of such specimens will greatly enhance the value and information of downstream applications. In this method, the specific cells are isolated on a microscope slide using photolithography, which will be faster, more specific, and less expensive than current methods. It relies on the fact that many biological molecules such as DNA are photosensitive and can be destroyed by ultraviolet irradiation. Therefore, it is possible to protect the contents of desired cells, yet destroy undesired cells. This approach leverages the technologies of the microelectronics industry, which can make features smaller than 1 micrometer with photolithography. A variety of ways has been created to achieve identification of the desired cell, and also to designate the other cells for destruction. This can be accomplished through chrome masks, direct laser writing, and also active masking using dynamic arrays. Image recognition is envisioned as one method for identifying cell nuclei and cell membranes. The pathologist can identify the cells of interest using a microscopic computerized image of the slide, and appropriate custom software. In one of the approaches described in this work, the software converts the selection into a digital mask that can be fed into a direct laser writer, e.g. the Heidelberg DWL66. Such a machine uses a metalized glass plate (with chrome metallization) on which there is a thin layer of photoresist. The laser transfers the digital mask onto the photoresist by direct writing, with typical best resolution of 2 micrometers. The plate is then developed to remove the exposed photoresist, which leaves the exposed areas susceptible to chemical chrome etch. The etch removes the unprotected chrome. The rest of the photoresist is then removed, by either ultraviolet organic solvent or over-development. The remaining chrome pattern is quickly oxidized by atmospheric exposure (typically within 30 seconds). The ready chrome mask is now applied to the tissue slide and aligned manually, or using automatic software and pre-designed alignment marks. The slide plate sandwich is then exposed to UV to destroy the DNA of the unwanted cells. The slide and plate are separated and the slide is processed in a standard way to prepare for polymerase chain reaction (PCR) and potential identification of cancer sequences.

Wade, Lawrence A.↗

International Space Station Carbon Dioxide Removal Assembly (ISS CDRA) Concepts and Advancements

An important aspect of air revitalization for life support in spacecraft is the removal of carbon dioxide from cabin air. Several types of carbon dioxide removal systems are in use in spacecraft life support. These systems rely on various removal techniques that employ different architectures and media for scrubbing CO2, such as permeable membranes, liquid amine, adsorbents, and absorbents. Sorbent systems have been used since the first manned missions. The current state of key technology is the existing International Space Station (ISS) Carbon Dioxide Removal Assembly (CDRA), a system that selectively removes carbon dioxide from the cabin atmosphere. The CDRA system was launched aboard UF-2 in February 2001 and resides in the U.S. Destiny Laboratory module. During the past four years, the CDRA system has operated with varying degrees of success. There have been several approaches to troubleshooting the CDRA system aimed at developing work-around solutions that would minimize the impact on astronaut time required to implement interim solutions. The paper discusses some of the short-term fixes applied to promote hardware life and restore functionality, as well as long-term plans and solutions for improving operability and reliability. The CDRA is a critical piece of life support equipment in the air revitalization system of the ISS, and is demonstrated technology that may ultimately prove well-suited for use in lunar or Mars base, and Mars transit life support applications.

ElSherif, Dina↗

Fabrication of Microshutter Arrays for James Webb Space Telescope

Two-dimensional MEMS microshutter arrays are being developed at NASA Goddard Space Flight Center for use in the near-infrared region on the Next Generation Space Telescope (NGST). The microshutter arrays are designed for the selective transmission of light with high efficiency and high contrast. The NGST environment requires cryogenic operation at 45K. Microshutter arrays are fabricated out of silicon-oxide-insulated (SOI) silicon wafers. Arrays are close-packed silicon nitride membranes with a pixel size of 100x100 microns. Individual shutters are patterned with a torsion flexure permitting shutters to open 90 degrees with a minimized mechanical stress concentration. The mechanical shutter arrays are fabricated using MEMS technologies. The processing includes a multi-layer metal deposition and patterning of shutter electrodes and magnetic pads, reactive ion etching (RE) of the front side to form shutters out of the nitride membrane, an anisotropic back-etch for wafer thinning, followed by a deep RIE (DRIE) back-etch down to the nitride shutter membrane to form frames and relieve shutters from the silicon substrate. An additional metal deposition and patterning is used to form back electrodes. Shutters are actuated using a magnetic force and latched using an electrostatic force.

Li, Mary J.↗