Auxin transport in geotropic curvatures of a branched plant.
Indole-acetic acid-2-C-14 movement in development of geotropic curvature in branches and immature plants from clone of Coleus blumei, Benth, studied by radioassay
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Indole-acetic acid-2-C-14 movement in development of geotropic curvature in branches and immature plants from clone of Coleus blumei, Benth, studied by radioassay
Radiobiology of Tradescantia clone orbited in Biosatellite 2, analyzing space effects on spontaneous and radiation induced mutation and cytological changes
Electron microscopical studies of a morphologically diverse, coccoid, presumably late Proterozoic blue-green alga are here reported. They show, together with light microscopy, that the form studied is widespread in the Cordilleran geosyncline, extend the record of well-defined endosporangia perhaps 700 million years into the past, and reveal previously unrecorded ultrastructural details. Coming from northeastern Utah, southwestern Alberta, and east central Alaska, these minute fossils belong to the recently described, morphologically diverse taxon Sphaerocongregus variabilis Moorman, are related to living entophysalidaceans, and have affinities with both the chroococcalean and chamaesiphonalean cyanophytes. Included in the morphological modes displayed by this alga are individual unicells, coenobial clusters of unicells, and a range of endosporangia comparable to those described for living entophysalidaceans. Scanning and transmission electron microscopy reveal that the endospores are commonly embedded in a vesicular matrix, that some of them show what appears to be a bilaminate or perhaps locally multilaminate wall structure, and that some remain together to mature as coenobial clones or 'colonies'. Taxonomic classification and phylogeny are discussed.
Immunomicrospheres are specially designed microscopic particles that have antibodies or similar molecules chemically bound to their surfaces. The antibody-coated microspheres react in a highly specific way with target cells, viruses, or other antigenic agents. Immunomicrospheres may be synthesized so that they incorporate compounds that are highly radioactive, intensely fluorescent, magnetic, electron opaque, highly colored, or pharmacologically active. These various types of microspheres may be coated with pure, highly specific monoclonal antibodies obtained by the new hybridoma cell cloning techniques or with conventional antibody preparations. Some of the many present and potential applications for these new reagents are (1) new types of radioimmune or immunofluorescent assays, (2) improved fluorescence microscopy, (3) separation of cells on the basis of the fluorescent, electrophoretic, or magnetic properties of bound immunomicrospheres, (4) markers for use in several types of electron or standard light microscopy, and (5) delivery of lethal compouds to specific undesirable living cells. The combination of the various new types of synthetic microspheres and the newly available homogeneous antibodies offers new opportunities in research, diagnosis, and therapy.
Recent evidence suggests that neurotrophic activity is required for the normal proliferation and development of muscle cells. The present paper reports a study of the purification and characterization of a neurotrophic factor (NTF) from adult chicken ischiatic-peroneal nerves using two independent quantitative in vitro assay systems. The assays were performed by the measurement of the incorporation of tritiated thymidine or the sizes of single-cell clones by chick muscle cells grown in culture. The greatest amount of neutrotrophic activity is found to be extracted at a pH of 8; aqueous suspensions of the activity are stable to long-term storage at room temperature. The specific activity of the substance is doubled upon precipitation with ammonium sulfate or after gel filtration, and increase 4 to 5 fold after salt gradient elution from DEAE cellulose columns. The active fraction obtained after gel filtration and rechromatography on DEAE cellulose exhibits a 7 to 10-fold increase in specific activity. Electrophoresis of the most highly purified material yields a greatly concentrated band at around 80,000 daltons. Although NTF is purified almost 10-fold as indicated by the increase in specific activity, the maximum activity of the partially purified material is greatly reduced, possibly due to a requirement for a cofactor for the expression of maximum activity.
A method for stimulating in vitro propagation of Guayule from a nutrient medium containing Guayule tissue by adding a substituted trialkyl amine bioinducing agent to the nutrient medium is described. Selective or differentiated propagation of shoots or callus is obtained by varying the amounts of substituted trialky amine present in the nutrient medium. The luxuriant growth provided may be processed for its poly isoprene content or may be transferred to a rooting medium for production of whole plants as identical clones of the original tissue. The method also provides for the production of large numbers of Guayule plants having identical desirable properties such as high polyisoprene levels.
Urokinase is a plasminogen activator found in urine. Relatively pure preparations have been tested in Europe, Japan and the United States for the treatment of deep vein thrombosis and other dangerous blood clots. Human embryonic kidney cell cultures have been found to produce urokinase at much higher concentrations, but less than 5% of the cells in typical cultures are producers. Since human diploid cells become senescent in culture the selection of clones derived from single cells will not provide enough material to be useful, so a bulk purification method is needed for the isolation of urokinase producing cell populations. Preparative cell electrophoresis was chosen as the method, since evidence exists that human embryonic cell cultures are richly heterogeneous with respect to electrophoretic mobility, and preliminary electrophoretic separations on the Apollo-Soyuz space flight produced cell populations that were rich in urokinase production. Similarly, erythropoietin is useful in the treatment of certain anemias and is a kidney cell duct, and electrophoretically enriched cell populations producing this product have been reported. Thus, there is a clear need for diploid human cells that produce these products, and there is evidence that such cells should be separable by free-flow cell electrophoresis.
Astronauts are provided much data and information via the monochrome CRT displays on the orbiter. For this project two areas were investigated for the possible introduction of computer graphics to enhance and extend the utility of these displays. One involved reviewing the current orbiter displays and identifying those which could be improved via computer graphics. As an example, the tabular data on electrical power distribution and control was enhanced by the addition of color and bar charts. The other dealt with the development of an aid to berthing a payload with the Remote Manipulator System (RMS). This aid consists of a graphics display of the top, front and side views of the payload and cargo bay and point of resolution (POR) position and attitude data for the current location of the payload. The initial implementation was on an IBM PC clone. The demonstration software installed in the Johnson Space Center Manipulator Development Facility (MD) was reviewed. Due to current hardware limitations, the MDF verision is slow, i.e., about a 40+ seond update rate and, hence, not real-time. Despite this fact, the evaluation of this additional visual cue as an RMS operator aid indicates that this display, with modifications for speed, etc., can assist the crew. Further development is appropriate.
Opportunities for releasing, capturing, constructing and/or fixing the differential expressions or response potentials of the higher plant genome in the hypo-g environment for commercialization are explored. General strategies include improved plant-growing, crop and forestry production systems which conserve soil, water, labor and energy resources, and nutritional partitioning and mobilization of nutrients and synthates. Tissue and cell culture techniques of commercial potential include the growing and manipulation of cultured plant cells in vitro in a bioreactor to produce biologicals and secondary plants of economic value. The facilitation of plant breeding, the cloning of specific pathogen-free materials, the elimination of growing point or apex viruses, and the increase of plant yield are other O-g applications. The space environment may be advantageous in somatic embryogenesis, the culture of alkaloids, and the development of completely new crop plant germ plasm.
A systematic photographic program, utilizing broad-band UV, blue, green, red and near IR pass-bands were carried out at the 60 cm. Tortugas Mountain Telescope. This is part of an ongoing program that spans two Jovian years (25 yrs). The purpose of this program is to provide an ongoing database to characterize high resolution data from spacecraft and earth-orbiting observatories. The standard observing program was maintained and support was given to other investigators who are carrying out complementary observing programs. The general aspect of the cloud decks were monitored, and reveal an interval that shows little variation in the general aspect of the clouddeck of Jupiter since 1981, when the North Temperature Belt darkened. A charge coupled device (CCD) camera was adapted for low cost operation, utilizing an IBM-AT clone for data acquisition. An RCA frame grabber was modified for encoding the archival images.
A large number of Intelligent Tutoring Systems (ITSs) have been built since they were first proposed in the early 1970's. Research conducted on the use of the best of these systems has demonstrated their effectiveness in tutoring in selected domains. A prototype ITS for tutoring students in the use of CLIPS language: CLIPSIT (CLIPS Intelligent Tutor) was developed. For an ITS to be widely accepted, not only must it be effective, flexible, and very responsive, it must also be capable of functioning on readily available computers. While most ITSs have been developed on powerful workstations, CLIPSIT is designed for use on the IBM PC/XT/AT personal computer family (and their clones). There are many issues to consider when developing an ITS on a personal computer such as the teaching strategy, user interface, knowledge representation, and program design methodology. Based on experiences in developing CLIPSIT, results on how to address some of these issues are reported and approaches are suggested for maintaining a powerful learning environment while delivering robust performance within the speed and memory constraints of the personal computer.
Any search for microbial life on Mars cannot rely upon cultivation of indigenous organisms. Only a minority of even terrestrial organisms that are observed in mixed, naturally-occurring microbial populations can be cultivated in the laboratory. Consequently, methods are being developed for analyzing the phylogenetic affiliations of the constituents of natural microbial populations without the need for their cultivation. This is more than an exercise in taxonomy, for the extent of phylogenetic relatedness between unknown and known organisms is some measure of the extent of their biochemical commonalities. In one approach, total DNA is isolated from natural microbial populations and 16S rRNA genes are shotgun cloned for rapid sequence determinations and phylogenetic analyses. A second approach employs oligodeoxynucleotide hybridization probes that bind to phylogenetic group-specific sequences in 16S rRNA. Since each actively growing cell contains about 104 ribosomes, the binding of the diagnostic probes to single cells can be visualized by radioactivity or fluorescence. The application of these methods and the use of in situ cultivation techniques is illustrated using submarine hydrothermal vent communities. Recommendations are made regarding planning toward future Mars missions.
It was proposed over 10 years ago that the archaebacteria represent a separate kingdom which diverged very early from the eubacteria and eukaryotes. It follows that investigations of archaebacterial characteristics might reveal features of early evolution. So far, two genes, one for bacteriorhodopsin and another for halorhodopsin, both from Halobacterium halobium, have been sequenced. We cloned and sequenced the gene coding for the polypeptide of another one of these rhodopsins, a halorhodopsin in Natronobacterium pharaonis. Peptide sequencing of cyanogen bromide fragments, and immuno-reactions of the protein and synthetic peptides derived from the C-terminal gene sequence, confirmed that the open reading frame was the structural gene for the pharaonis halorhodopsin polypeptide. The flanking DNA sequences of this gene, as well as those of other bacterial rhodopsins, were compared to previously proposed archaebacterial consensus sequences. In pairwise comparisons of the open reading frame with DNA sequences for bacterio-opsin and halo-opsin from Halobacterium halobium, silent divergences were calculated. These indicate very considerable evolutionary distance between each pair of genes, even in the dame organism. In spite of this, three protein sequences show extensive similarities, indicating strong selective pressures.
Ongoing work to test the hypothesis of the origin of eukaryotic cell organelles by microbial symbioses is discussed. Because of the widespread acceptance of the serial endosymbiotic theory (SET) of the origin of plastids and mitochondria, the idea of the symbiotic origin of the centrioles and axonemes for spirochete bacteria motility symbiosis was tested. Intracellular microtubular systems are purported to derive from symbiotic associations between ancestral eukaryotic cells and motile bacteria. Four lines of approach to this problem are being pursued: (1) cloning the gene of a tubulin-like protein discovered in Spirocheata bajacaliforniesis; (2) seeking axoneme proteins in spirochets by antibody cross-reaction; (3) attempting to cultivate larger, free-living spirochetes; and (4) studying in detail spirochetes (e.g., Cristispira) symbiotic with marine animals. Other aspects of the investigation are presented.
Experiment critical use of RS-232 data busses in the Transport Systems Research Vehicle (TSRV) operated by the Advanced Transport Operating Systems Program Office at the NASA Langley Research Center has recently increased. Each application utilizes a number of nonidentical computer and peripheral configurations and requires task specific software development. To aid these development tasks, an IBM PC-based RS-232 bus monitoring system was produced. It can simultaneously monitor two communication ports of a PC or clone, including the nonstandard bus expansion of the TSRV Grid laptop computers. Display occurs in a separate window for each port's input with binary display being selectable. A number of other features including binary log files, screen capture to files, and a full range of communication parameters are provided.
A recent upgrade of the Transport Systems Research Vehicle (TSRV) operated by the Advanced Transport Operating Systems Program Office at the NASA Langley Research Center included installation of a number of Grid 1500 series laptop computers. Each unit is a 80386-based IBM PC clone. RS-232 data busses are needed for TSRV flight research programs, and it has been advantageous to extend the application of the Grids in this area. Use was made of the expansion features of the Grid internal bus to add a user programmable serial communication channel. Software to allow use of the Grid bus expansion has been written and placed in a Turbo C library for incorporation into applications programs in a transparent manner via function calls. Port setup; interrupt-driven, two-way data transfer; and software flow control are built into the library functions.
The OspA gene from a North American strain of the Lyme disease Spirochete, Borrelia burgdorferi, was cloned under the control of transciption and translation signals from bacteriophage T7. Full-length OspA protein, a 273 amino acid (31kD) lipoprotein, is expressed poorly in Escherichia coli and is associated with the insoluble membrane fraction. In contrast, a truncated form of OspA lacking the amino-terminal signal sequence which normally would direct localization of the protein to the outer membrane is expressed at very high levels (less than or equal to 100 mg/liter) and is soluble. The truncated protein was purified to homogeneity and is being tested to see if it will be useful as an immunogen in a vaccine against Lyme disease. Circular dichroism and fluorescence spectroscopy was used to characterize the secondary structure and study conformational changes in the protein. Studies underway with other surface proteins from B burgdorferi and a related spirochete, B. hermsii, which causes relapsing fever, leads us to conclude that a strategy similar to that used to express the truncated OspA can provide a facile method for producing variations of Borrelia lipoproteins which are highly expressed in E. coli and soluble without exposure to detergents.
The reason why Titan has a thick atmosphere while its Jovian clones Ganymede and Callisto have nothing is addressed. It is argued that this contrast is a predictable consequence of impact erosion, i.e., the escape of atmospheric gases as a consequence of hypervelocity impact, the average impact velocity of stray bodies on Titan being lower than on Ganymede and Callisto. A model to examine this hypothesis is presented.