ALBERT

Description: A three-dimensional (3-D), tissue-like model was developed for the genotoxic assessment of space environment. In previous experiments, we found that culturing mammalian cells in a NASA-designed bioreactor, using Cytodex-3 beads as a scaffold, generated 3-D multicellular spheroids. In an effort to generate scaffold-free spheroids, we developed a new 3-D tissue-like model by coculturing fibroblast and epithelial cell in a NASA bioreactor using macroporous Cultispher-S(TradeMark) microcarriers. Big Blue(Registered Trademark) Rat 2(Lambda) fibroblasts, genetically engineered to contain multiple copies (〉60 copies/cell) of the Lac I target gene, were cocultured with radio-sensitive human epithelial cells, H184F5. Over an 8-day period, samples were periodically examined by microscopy and histology to confirm cell attachment, growth, and viability. Immunohistochemistry and western analysis were used to evaluate the expression of specific cytoskeletal and adhesion proteins. Key cell culture parameters (glucose, pH, and lactate concentrations) were monitored daily. Controls were two-dimensional mono layers of fibroblast or epithelial cells cultured in T-flasks. Analysis of 3-D spheroids from the bioreactor suggests fibroblast cells attached to and completely covered the bead surface and inner channels by day 3 in the bioreactor. Treatment of the 3-day spheroids with dispase II dissolved the Cultisphers(TradeMark) and produced multicellular, bead-less constructs. Immunohistochemistry confirmed the presence of vi.mentin, cytokeratin and E-cadherin in treated spheroids. Examination of the dispase II treated spheroids with transmission electron microscopy (TEM) also showed the presence of desmosomes. These results suggest that the controlled enzymatic degradation of an artificial matrix in the low shear environment of the NASA-designed bioreactor can produce 3-D tissue-like spheroids. 2

Description: As part of a project to complete a comprehensive catalogue of astrophysically relevant emission lines in support of new-generation X-ray observatories using the Lawrence Livermore electron beam ion traps EBIT-I and EDIT-II, emission lines of argon and sulfur in the soft X-ray and extreme ultraviolet region were studied. Observations of Ar IX through Ar XVI and S VII through S XIV between 20 and 75 Angstrom are presented to illustrate our work.

Description: Using the Moon to occult the Sun, the Clementine spacecraft used its navigation cameras to map the inner zodiacal light at optical wavelengths over elongations of 3 approx. less than epsilon approx. less than 30 deg from the Sun. This surface brightness map is then used to infer the spatial distribution of interplanetary dust over heliocentric distances of about 10 solar radii to the orbit of Venus. The averaged ecliptic surface brightness of the zodiacal light falls off as Z(epsilon) is a member of epsilon(sup -2.45 +/- 0.05), which suggests that the dust cross-sectional density nominally falls off as sigma(r) is a member of r(sup - 1.45 +/- 0.05). The interplanetary dust also has an albedo of alpha approx. = 0.1 that is uncertain by a factor of approx. 2. Asymmetries of approx. 10% are seen in directions east-west and north-south of the Sun, and these may be due the giant planets' secular gravitational perturbations. We apply a simple model that attributes the zodiacal light as due to three dust populations having distinct inclination distributions, namely, dust from asteroids and Jupiter-family comets (JFCs) having characteristic inclinations of i approx. 7 deg, dust from Halley-type comets having i approx. 33 deg, and an isotropic cloud of dust from Oort Cloud comets. The best-fitting scenario indicates that asteroids + JFCs are the source of about 45% of the optical dust cross section seen in the ecliptic at 1 AU but that at least 89% of the dust cross section enclosed by a 1-AU-radius sphere is of a cometary origin. Each population's radial density variations can also deviate somewhat from the nominal sigma(r) is a member of r(sup -1.45). When these results are extrapolated out to the asteroid belt, we find an upper limit on the mass of the light-reflecting asteroidal dust that is equivalent to a 12-km asteroid, and a similar extrapolation of the isotropic dust cloud out to Oort Cloud distances yields a mass equivalent to a 30-km comet, although the latter mass is uncertain by orders of magnitude.

Description: The program "Nanotechnology Infrared Optics for Astronomy Missions" will design and develop new, nanotechnology techniques for infrared optical devices suitable for use in NASA space missions. The proposal combines expertise from the Smithsonian Astrophysical Observatory, the Naval Research Laboratory, the Goddard Space Flight Center, and the Physics Department at the Queen Mary and Westfield College in London, now relocated to the University of Cardiff, Cardiff, Wales. The method uses individually tailored metal grids, and layered stacks of metal mesh grids, both inductive (free-standing) and capacitive (substrate-mounted), to produce various kinds of filters. The program has the following goals: (1) Model FIR filter properties using electric-circuit analogs, and near-field, EM diffraction calculations; (2) Prototype fabrication of meshes on various substrates, with various materials, and of various dimensions; (3) Test of filter prototypes, and iterate with the modeling programs; (4) Travel to related sites, including trips to Washington, D.C. (location of NRL and GSFC), London (location of QMW), Cardiff, Wales, and Rome (location of ISO PMS project headquarters); (5) Produce ancillary science, including publication of both testing on mesh performance and infrared astronomical science.

Description: We completed all major fabrication and testing for the third telescope and phase-closure operation at the Infrared-Optical Telescope Array (IOTA) during this period. In particular we successfully tested the phase-closure operation, using a laboratory light source illuminating the full delay-line optical paths, and using an integrated-optic beam combiner coupled to our Picnic-detector camera. This demonstration is an important and near-final milestone achievement. As of this writing, however, several tasks yet remain, owing to development snags and weather, so the final proof of success, phase-closure observation of a star, is now expected to occur in early 2002, soon after this report has been submitted.

Description: The idea behind genetic algorithms is to extract optimization strategies nature uses successfully - known as Darwinian Evolution - and transform them for application in mathematical optimization theory to find the global optimum in a defined phase space. One could imagine a population of individual 'explorers' sent into the optimization phase-space. Each explorer is defined by its genes, what means, its position inside the phase-space is coded in his genes. Every explorer has the duty to find a value of the quality of his position in the phase space. (Consider the phase-space being a number of variables in some technological process, the value of quality of any position in the phase space - in other words: any set of the variables - can be expressed by the yield of the desired chemical product.) Then the struggle of 'life' begins. The three fundamental principles are selection, mating/crossover, and mutation. Only explorers (= genes) sitting on the best places will reproduce and create a new population. This is performed in the second step (mating/crossover). The 'hope' behind this part of the algorithm is, that 'good' sections of two parents will be recombined to yet better fitting children. In fact, many of the created children will not be successful (as in biological evolution), but a few children will indeed fulfill this hope. These good sections are named in some publications as building blocks. Now there appears a problem. Repeating these steps, no new area would be explored. The two former steps would only exploit the already known regions in the phase space, which could lead to premature convergence of the algorithm with the consequence of missing the global optimum by exploiting some local optimum. The third step, mutation, ensures the necessary accidental effects. One can imagine the new population being mixed up a little bit to bring some new information into this set of genes. Whereas in biology a gene is described as a macro-molecule with four different bases to code the genetic information, a gene in genetic algorithms is usually defined as a bitstring (a sequence of b 1's and 0's).

Description: In vitro evolution methods were used to obtain DNA enzymes that cleave either a 2',5' - phosphodiester following a wibonucleotide or a 3',5' -phosphodiester following an L-ribonucleotide. Both enzymes can operate in an intermolecular reaction format with multiple turnover. The DNA enzyme that cleaves a 2',5' -phosphodiester exhibits a k(sub cat) of approx. 0.01/ min and catalytic efficiency, k(sub cat)/k(sub m) of approx. 10(exp 5)/ M min. The enzyme that cleaves an L-ribonudeotide is about 10-fold slower and has a catalytic efficiency of approx. 4 x 10(exp 5)/ M min. Both enzymes require a divalent metal cation for their activity and have optimal catalytic rate at pH 7-8 and 35-50 C. In a comparison of each enzyme s activity with either its corresponding substrate that contains an unnatural ribonudeotide or a substrate that instead contains a standard ribonucleotide, the 2',5' -phosphodiester-deaving DNA enzyme exhibited a regioselectivity of 6000- fold, while the L-ribonucleotide-cleaving DNA enzyme exhibited an enantioselectivity of 50-fold. These molecules demonstrate how in vitro evolution can be used to obtain regio- and enantioselective catalysts that exhibit specificities for nonnatural analogues of biological compounds.

Description: Multifiber optical spectroscopy has been performed on galaxies in the vicinity of strong, nearby radio galaxies. These radio galaxies were selected from the, 3CR and B2 catalogs based on their exclusion from the Abell catalog, which is puzzling given the hypothesis that an external medium is required to confine the radio plasma of such galaxies. Velocities derived from the spectra were used to confirm the existence of groups and poor clusters in the fields of most of the radio galaxies. We find that all radio galaxies with classical Fanaroff-Riley type I morphologies prove to reside in clusters, whereas the other radio galaxies often appear to be recent galaxy-galaxy mergers in regions of low galaxy density. These findings confirm the earlier result that the existence of extended X-ray emission combined with a statistical excess of neighboring galaxies can be used to identify poor clusters associated with radio galaxies.

Description: The primary purpose of this project was to evaluate and improve models for the bulk-skin temperature difference to the point where they could accurately and reliably apply under a wide variety of environmental conditions. To accomplish this goal, work was conducted in three primary areas. These included production of an archive of available data sets containing measurements of the skin and bulk temperatures and associated environmental conditions, evaluation of existing skin layer models using the compiled data archive, and additional theoretical work on the development of an improved model using the data collected under diverse environmental conditions. In this work we set the basis for a new physical model of renewal type, and propose a parameterization for the temperature difference across the cool skin of the ocean in which the effects of thermal buoyancy, wind stress, and microscale breaking are all integrated by means of the appropriate renewal time scales. Ideally, we seek to obtain a model that will accurately apply under a wide variety of environmental conditions. A summary of the work in each of these areas is included in this report. A large amount of work was accomplished under the support of this grant. The grant supported the graduate studies of Sandra Castro and the preparation of her thesis which will be completed later this year. This work led to poster presentations at the 1999 American Geophysical Union Fall Meeting and 2000 IGARSS meeting. Additional work will be presented in a talk at this year's American Meteorological Society Air-Sea Interaction Meeting this May. The grant also supported Sandra Castro during a two week experiment aboard the R/P Flip (led by Dr. Andrew Jessup of the Applied Physics Laboratory) to help obtain additional shared data sets and to provide Sandra with a fundamental understanding of the physical processes needed in the models. In a related area, the funding also partially supported Dr. William Emery and Daniel Baldwin in the preparation of their publication "Accuracy of in situ sea surface temperatures used to calibrate infrared satellite measurements". The remainder of this report is drawn from these publications and presentations.

Description: Thin films of an organic nonlinear optical (NLO) material, N, N-dimethyl-p-(2,2-dicyanovinyl) aniline (DCVA), have been grown in space and on the ground by physical vapor transport in an effusive ampoule arrangement. The thin film growth technique developed on the ground is a direct result of information gleaned from experiments in microgravity. This paper covers the results of our experimental investigations for establishing "ideal" terrestrial conditions for deposition of a DCVA film. The active control during the deposition process was exercised by three deposition variables: the material source temperature, the background pressure external to the growth ampoule and the substrate temperature. Successful growth occurred when the difference in temperature between the source material and the copper substrate was 14 degrees C and the background nitrogen pressure was such that the transport was either diffusive or convective. A qualitative diffusion limited boundary was estimated to occur at a pressure of approximately 20 torr. We have probed the DCVA thin films with visible-near infrared reflection absorption spectroscopy, polarized Fourier transform infrared spectrometry, differential interference contrast optical microscopy, and stylus profilometry.