ALBERT

Description: A mesophilic toluene-degrading consortium (TDC) and an ethylbenzene-degrading consortium (EDC) were established under sulfate-reducing conditions. These consortia were first characterized by denaturing gradient gel electrophoresis (DGGE) fingerprinting of PCR-amplified 16S rRNA gene fragments, followed by sequencing. The sequences of the major bands (T-1 and E-2) belonging to TDC and EDC, respectively, were affiliated with the family Desulfobacteriaceae. Another major band from EDC (E-1) was related to an uncultured non-sulfate-reducing soil bacterium. Oligonucleotide probes specific for the 16S rRNAs of target organisms corresponding to T-1, E-1, and E-2 were designed, and hybridization conditions were optimized for two analytical formats, membrane and DNA microarray hybridization. Both formats were used to characterize the TDC and EDC, and the results of both were consistent with DGGE analysis. In order to assess the utility of the microarray format for analysis of environmental samples, oil-contaminated sediments from the coast of Kuwait were analyzed. The DNA microarray successfully detected bacterial nucleic acids from these samples, but probes targeting specific groups of sulfate-reducing bacteria did not give positive signals. The results of this study demonstrate the limitations and the potential utility of DNA microarrays for microbial community analysis.

Description: Oligonucleotide microarrays were used to profile directly extracted rRNA from environmental microbial populations without PCR amplification. In our initial inspection of two distinct estuarine study sites, the hybridization patterns were reproducible and varied between estuarine sediments of differing salinities. The determination of a thermal dissociation curve (i.e., melting profile) for each probe-target duplex provided information on hybridization specificity, which is essential for confirming adequate discrimination between target and nontarget sequences.

Description: The discrimination between perfect-match and single-base-pair-mismatched nucleic acid duplexes was investigated by using oligonucleotide DNA microarrays and nonequilibrium dissociation rates (melting profiles). DNA and RNA versions of two synthetic targets corresponding to the 16S rRNA sequences of Staphylococcus epidermidis (38 nucleotides) and Nitrosomonas eutropha (39 nucleotides) were hybridized to perfect-match probes (18-mer and 19-mer) and to a set of probes having all possible single-base-pair mismatches. The melting profiles of all probe-target duplexes were determined in parallel by using an imposed temperature step gradient. We derived an optimum wash temperature for each probe and target by using a simple formula to calculate a discrimination index for each temperature of the step gradient. This optimum corresponded to the output of an independent analysis using a customized neural network program. These results together provide an experimental and analytical framework for optimizing mismatch discrimination among all probes on a DNA microarray.

Description: The effects of single-base-pair near-terminal and terminal mismatches on the dissociation temperature (T(d)) and signal intensity of short DNA duplexes were determined by using oligonucleotide microarrays and neural network (NN) analyses. Two perfect-match probes and 29 probes having a single-base-pair mismatch at positions 1 to 5 from the 5' terminus of the probe were designed to target one of two short sequences representing 16S rRNA. Nonequilibrium dissociation rates (i.e., melting profiles) of all probe-target duplexes were determined simultaneously. Analysis of variance revealed that position of the mismatch, type of mismatch, and formamide concentration significantly affected the T(d) and signal intensity. Increasing the concentration of formamide in the washing buffer decreased the T(d) and signal intensity, and it decreased the variability of the signal. Although T(d)s of probe-target duplexes with mismatches in the first or second position were not significantly different from one another, duplexes with mismatches in the third to fifth positions had significantly lower T(d)s than those with mismatches in the first or second position. The trained NNs predicted the T(d) with high accuracies (R(2) = 0.93). However, the NNs predicted the signal intensity only moderately accurately (R(2) = 0.67), presumably due to increased noise in the signal intensity at low formamide concentrations. Sensitivity analysis revealed that the concentration of formamide explained most (75%) of the variability in T(d)s, followed by position of the mismatch (19%) and type of mismatch (6%). The results suggest that position of the mismatch at or near the 5' terminus plays a greater role in determining the T(d) and signal intensity of duplexes than the type of mismatch.

Description: Science and technology development from balloon-borne telescopes and experiments is a rich return on a relatively modest involvement of NASA resources. For the past three decades, the development of increasingly competitive and complex science payloads and observational programs from high altitude balloon-borne platforms has yielded significant scientific discoveries. The success and capabilities of scientific balloons are closely related to advancements in the textile and plastic industries. This paper will present an overview of scientific balloons as a viable and economical platform for transporting large telescopes and scientific instruments to the upper atmosphere to conduct scientific missions. Additionally, the paper sheds the light on the problems associated with UV degradation of high performance textile components that are used to support the payload of the balloon and proposes future research to reduce/eliminate Ultra Violet (UV) degradation in order to conduct long-term scientific missions.

Description: For the last three decades, NASA has been involved in the development of giant balloons that are capable of lifting heavy payloads of equipment (such as large telescopes and scientific instruments) to the upper atmosphere. While the use of such balloons has led to scientific discoveries, the demand for competitive science payloads and observational programs continues to rise. The NASA Balloon Program Office has entered a new phase of research to develop an Ultra Long Duration Balloon (ULDB) that will lift payloads of up to 3,600 kg to altitudes of up to 40 km. The flight duration is targeted to ranges between 30 to 100 days. Attaining these target durations requires the development of a super-pressure balloon design. The use of textile structures have already been established in these missions in the form of high strength tendons essential for the super pressure pumpkin design. Unfortunately, high strength fibers lose significant strength upon exposure to Ultra Violet (UV) radiation. Such UV degradation poses a serious challenge for the development of the ULDB. To improve the mission performance of the ULDB, new methods for protecting the tendons from the environmental effects need to be developed. NASA and NC State University College of Textiles are undertaking a research program to address these issues. Four tracks have been identified to prepare finishes that are believed to enhance the resistance of high strength fibers to UV. These tracks are: (a) self-polymerizing, (b) diffusion application, (c) polymer-filled with 30-40% UV absorber, and (d) combination of dyeing plus surface application. Four high performance fibers have been selected for this research investigation. These are Vectran (trademark), Spectra (trademark), Kevlar (trademark) and, PBO (Zylon (trademark)). This work will address the current progress of evaluating the performance of the UV finishes. This will be accomplished by comparing the tensile properties (strength, breaking elongation, modulus, etc) of untreated, unexposed to UV fibers; untreated exposed to UV fibers; and treated exposed to UV fibers.

Description: The development and characterization of materials suitable for ultra long duration balloon flights has recently been the focus of the materials R&D efforts for the NASA balloon program. Although basic materials selection criteria is similar to those used for conventional balloon missions, additional considerations related to balloon design, fabrication, durability, environmental effects, and cost must also be considered. Among these, the highest impact on material selection is, related to the design shape of the balloon. Work done by independent researchers indicate that for pumpkin type balloons, the load-carrying member is the tendon rather than the structural envelope. This in turn lowers the strength requirements on the envelope material to a large extent. Several materials and material combinations were explored for both design systems. This paper will present the progress made to date in the selection and characterization of these materials and the technical challenges remaining to be overcome,

Description: This document contains a collection of experiments presented and demonstrated at the National Educators' Workshop: Update 2001 held in College Park, Maryland, October 14-17, 2001.

Description: This report describes the activities completed under a grant from the NASA Langley Research Center to develop a plan for the assessment, improvement, and deployment of a Radar Acoustic Sounding System (RASS) for the detection of wake vortices. A brief review is provided of existing alternative instruments for wake vortex detection. This is followed by a review of previous implementations and assessment of a RASS. As a result of this review, it is concluded that the basic features of a RASS have several advantages over other commonly used wake vortex detection and measurement systems. Most important of these features are the good fidelity of the measurements and the potential for all weather operation. To realize the full potential of this remote sensing instrument, a plan for the development of a RASS designed specifically for wake vortex detection and measurement has been prepared. To keep costs to a minimum, this program would start with the development an inexpensive laboratory-scale version of a RASS system. The new instrument would be developed in several stages, each allowing for a critical assessment of the instrument s potential and limitations. The instrument, in its initial stages of development, would be tested in a controlled laboratory environment. A jet vortex simulator, a prototype version of which has already been fabricated, would be interrogated by the RASS system. The details of the laboratory vortex would be measured using a Particle Image Velocimetry (PIV) system. In the early development stages, the scattered radar signal would be digitized and the signal post-processed to determine how extensively and accurately the RASS could measure properties of the wake vortex. If the initial tests prove to be successful, a real-time, digital signal processing system would be developed as a component of the RASS system. At each stage of the instrument development and testing, the implications of the scaling required for a full-scale instrument would be considered. It is concluded that a RASS system, developed for the specific application of wake vortex detection, could become part of a robust Aircraft Vortex Spacing System (AVOSS). This system, in turn, could contribute to Reduced Spacing Operations (RSO) in US airports and improvements in Terminal Area productivity (TAP).

Description: Substance P inhibits renin secretion. This polypeptide is a transmitter in primary afferent neurons and is released from the peripheral as well as the central portions of these neurons. It is present in afferent nerves from the kidneys. Neuropeptide Y, which is a cotransmitter with norepinephrine and epinephrine, is found in sympathetic neurons that are closely associated with and presumably innervate the juxtagolmerular cells. Its effect on renin secretion is unknown, but it produces renal vasoconstriction and natriuresis. Vasoactive intestinal polypeptide (VIP) is a cotransmitter with acetylocholine in cholinergic neurons, and this polypeptide stimulates renin secretion. We cannot find any evidence for its occurence in neurons in the kidneys, but various stimuli increase plasma VIP to levels comparable to those produced by doses of exogenous VIP which stimulated renin secretion. Neostigmine increases plasma VIP and plasma renin activity, and the VIP appears to be responsible for the increase in renin secretion, since the increase is not blocked by renal denervation or propranolol. Stimulation of various areas in the brain produces sympathetically mediated increases in plasma renin activity associated with increases in blood pressure. However, there is pharmacological evidence that the renin response can be separated from the blood pressure response. In anaesthetized dogs, drugs that increase central serotonergic discharge increase renin secretion without increasing blood pressure. In rats, activation of sertonergic neurons in the dorsal raphe nucleus increases renin secretion by a pathway that projects from this nucleus to the ventral hypothalamus, and from there to the kidneys via the sympathetic nervous system. The serotonin releasing drug parachloramphetamine also increases plasma VIP, but VIP does not appear to be the primary mediator of the renin response. There is preliminary evidence that the serotonergic neurons in the dorsal raphe nucleus are part of the pathway by which psychosocial stimuli increase renin secretion.