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At least 163 records · Page 9

Non-invasive optical detection of glucose in cell culture nutrient medium

The objective of the proposed research was to begin the development of a non-invasive optical sensor for measuring glucose concentration in the output medium of cell cultures grown in a unique NASA bioreactor referred to as an integrated rotating-wall vessel (IRWV). The input, a bovine serum based nutrient media, has a known glucose concentration. The cells within the bioreactor digest a portion of the glucose. Thus, the non-invasive optical sensor is needed to monitor the decrease in glucose due to cellular consumption since the critical parameters for sustained cellular productivity are glucose and pH. Previous glucose sensing techniques have used chemical reactions to quantify the glucose concentration. Chemical reactions, however, cannot provide for continuous, real time, non-invasive measurement as is required in this application. Our effort while in the fellowship program was focused on the design, optical setup, and testing of one bench top prototype non-invasive optical sensor using a mid-infrared absorption spectroscopy technique. Glucose has a fundamental vibrational absorption peak in the mid-infrared wavelength range at 9.6 micron. Preliminary absorption data using a CO2 laser were collected at this wavelength for water based glucose solutions at different concentrations and one bovine serum based nutrient medium (GTSF) with added glucose. The results showed near linear absorption responses for the glucose-in-water data with resolutions as high at 108 mg/dl and as low as 10 mg/dl. The nutrient medium had a resolution of 291 mg/dl. The variability of the results was due mainly to thermal and polarization drifts of the laser while the decrease in sensitivity to glucose in the nutrient medium was expected due to the increase in the number of confounders present in the nutrient medium. A multispectral approach needs to be used to compensate for these confounders. The CO2 laser used for these studies was wavelength tunable (9.2 to 10.8 micrometers), however, it was to unstable across wavelengths to test the multispectral approach. From this research, further NASA support was obtained to continue the work throughout the year in which a more stable light source will be used at smaller, near-infrared, wavelengths. It is anticipated that a more compact, non-invasive, optical glucose sensor will be realized which can be used with a bioreactor on future space shuttle missions. It is also anticipated that a multispectral optical sensor may be used to determine the concentration of other molecules needed within the NASA bioreactor, such as fructose and galactose.

Cote, Gerald L.↗

ICE/ISEE plasma wave data analysis

This report is one of the final processing of ICE plasma wave (pw) data and analysis of late ISEE 3, ICE cometary, and ICE cruise trajectory data, where coronal mass ejections (CME's) were the first locus of attention. Interest in CME's inspired an effort to represent our pw data in a condensed spectrogram format that facilitated rapid digestion of interplanetary phenomena on long (greater than 1 day) time scales. The format serendipitously allowed us to also examine earth-orbiting data from a new perspective, invigorating older areas of investigation in Earth's immediate environment. We, therefore, continued to examine with great interest the last year of ISEE 3's precomet phase, when it spent considerable time far downwind from Earth, recording for days on end conditions upstream, downstream, and across the very weak, distant flank bow shock. Among other motivations has been the apparent similarity of some shock and post shock structures to the signatures of the bow wave surrounding comet Giacobini-Zinner, whose ICE-phase data we revisited.

Greenstadt, E. W.↗

Applying Virtual Reality to commercial Edutainment

Virtual reality (VR) when defined as a computer generated, immersive, three dimensional graphics environment which provides varying degrees of interactivity, remains an expensive, highly specialized application, yet to find its way into the school, home, or business. As a novel approach to a theme park-type attraction, though, its use can be justified. This paper describes how a virtual reality 'tour of the human digestive system' was created for the Omniplex Science Museum of Oklahoma City, Oklahoma. The customers main objectives were: (1) to educate; (2) to entertain; (3) to draw visitors; and (4) to generate revenue. The 'Edutainment' system ultimately delivered met these goals. As more such systems come into existence the resulting library of licensable programs will greatly reduce development costs to individual institutions.

Grissom, F.↗

Applications catalog of pyrotechnically actuated devices/systems

A compilation of basic information on pyrotechnically actuated devices/systems used in NASA aerospace and aeronautic applications was formatted into a catalog. The intent is to provide (1) a quick reference digest of the types of operational pyro mechanisms and (2) a source of contacts for further details. Data on these items was furnished by the NASA Centers that developed and/or utilized such devices to perform specific functions on spacecraft, launch vehicles, aircraft, and ground support equipment. Information entries include an item title, user center name, commercial contractor/vendor, identifying part number(s), a basic figure, briefly described purpose and operation, previous usage, and operational limits/requirements.

Seeholzer, Thomas L.↗

Regional differences in lectin binding patterns of vestibular hair cells

Surface glycoconjugates of hair cells and supporting cells in the vestibular endorgans of the bullfrog were identified using biotinylated lectins with different carbohydrate specificities. Lectin binding in hair cells was consistent with the presence of glucose and mannose (CON A), galactose (RCA-I), N-acetylgalactosamine (VVA), but not fucose (UEA-I) residues. Hair cells in the bullfrog sacculus, unlike those in the utriculus and semicircular canals, did not stain for N-acetylglucosamine (WGA) or N-acetylgalactosamine (VVA). By contrast, WGA and, to a lesser extent, VVA, differentially stained utricular and semicircular canal hair cells, labeling hair cells located in peripheral, but not central, regions. In mammals, WGA uniformly labeled Type 1 hair cells while labeling, as in the bullfrog, Type 2 hair cells only in peripheral regions. These regional variations were retained after enzymatic digestion. We conclude that vestibular hair cells differ in their surface glycoconjugates and that differences in lectin binding patterns can be used to identify hair cell types and to infer the epithelial origin of isolated vestibular hair cells.

Baird, Richard A.↗

Studies on the Mechanisms of Microbial Adaptation to the Physical Environment

The environmental factors which affect humans and other animals also influence the microorganisms which are such an important part of our ecology. Some of the microorganisms are very closely associated with animals, living in the digestive tract and synthesizing essential nutrients for the host. For these microbes, most external physical changes are of little consequence, because they are well shielded by the animals' homeostatic systems. The vast majority of microorganisms, however, live free in nature, especially in the soil and oceans. It has been estimated that the upper 15 cm of a fertile soil may contain over 4000 kg of bacteria and fungi per hectare. These organisms are responsible for degrading the complex molecules of plants and animals when they die, eventually producing simple organics, carbon dioxide, and inorganics, which are then used for the next cycle of plant growth. It is believed that over 90 % of the biologically produced carbon dioxide results from the metabolic activity of bacteria and fungi. In addition to recycling plant nutrients, soil bacteria also provide new nutrients through 'fixation' of atmospheric nitrogen into ammonia and nitrate, the forms which can be used by plants. Microorganisms so have an enormous capacity for detoxifying both natural and man-made poisons. All of these functions of microorganisms are essential to the operation of the material cycles on Earth. This is true of all locations on the planet, regardless of the climate or other environmental factors. In fact, one of the most impressive attributes of microorganisms is their ability to adapt to every stable environment on Earth. These include such extremes as polar regions, hot springs, water saturated with salt, mountain tops, ocean depths, acid and alkaline waters, deserts, intense radioactivity, soil and water contaminated with toxic chemicals or petroleum, and areas devoid of oxygen.

Heinrich, M. R.↗

Rubidium-Strontium Formation Age of Allan Hills 84001 Carbonates

Our preferred age for the formation of carbonates in the martian meteorite ALH 84001 is 3.90 plus or minus 0.04 Ga for Lambda (Rubidium 87) equals 0.01402 Ga (exp -1), or 3.85 plus or minus 0.04 Ga for Lambda (Rubidium 87) = 0.0142 Ga(exponent -1). This age is determined by a three-point Rb-Sr isochron defined by leachates of high-graded carbonate-rich material. Major cation and especially phosphorous analyses of the leachates permit contributions from igneous whitlockite to be recognized for low-acidity leachates, and the corresponding data are omitted from the isochron. Data for the two highest acidity leachates plot close to the preferred isochron, but are omitted because we believe they contain contributions leached from the pyroxene substrate on which most of the carbonates are found. Nevertheless, the isochron age for all five highest-acidity leachates is 3.94 plus or minus 0.04 Ga, and is within error of the age obtained for the more restricted data set. All leachates used to define the isochron have major cation compositions that are singular to those obtained by microprobe analyses of the carbonate rosettes and are consistent with progressive digestion of the carbonates according to their composition. The age thus obtained for the carbonates is about 600 m.y. younger than the crystalization age of ALH 84001 determined by Sm-Nd analyses but is within error limits of the age of impact metamorphism inferred from the Rb-Sr and Ar-Ar systematics of silicates. which yield ages of 3.85 plus or minus 0.05 Ga and 4.05- 3.80 Ga to 4.3-3.8 Ga, respectively. Similarities between the carbonate crystallization age and the age of impact metamorphism as determined by Ar-Ar and Rb-Sr suggest that the carbonate formation is impact-related. Nevertheless, both high and low- temperature scenarios for the origin of the carbonates are possible.

Borg, L. E.↗

Lamb Wave Assessment of Fiber Volume Fraction in Composites

Among the various techniques available, ultrasonic Lamb waves offer a convenient method of examining composite materials. Since the Lamb wave velocity depends on the elastic properties of a material, an effective tool exists to evaluate composites by measuring the velocity of these waves. Lamb waves can propagate over long distances and are sensitive to the desired in-plane elastic properties of the material. This paper discusses a study in which Lamb waves were used to examine fiber volume fraction variations of approximately 0.40-0.70 in composites. The Lamb wave measurements were compared to fiber volume fractions obtained from acid digestion tests. Additionally, a model to predict the fiber volume fraction from Lamb wave velocity values was evaluated.

Seale, Michael D.↗

Effects of Microgravity on Embryonic Quail Eye Development

Immunohistochemical methods were used to stain neurofilament protein in corneal nerves of Embryonic Day 16 (E16) quail eyes that had been fixed in 4% paraformaldehyde at room temperature for several months. Fixation was according to the methods used by the Mir 21/NASA 2 Avian Developmental Biology Flight Experiments for quail embryos incubated on the Mir Space Station. After fixation, corneas were pretreated to improve immunohistochemical visualization of neurofilaments. A sequential combination of three pretreatments [microwave heating in saline G, followed by extraction with sodium dodecyl sulfate (SDS) at 37 C, followed by digestion with hyaluronidase at 37 C], produced increased antibody staining of corneal nerve neurofilament proteins, compared with corneas subjected to no prior pretreatments. Darker nerve staining and increased numbers of fine branches were observed, together with lower background staining after such pretreatments. In contrast, use of any single pretreatment or pair of pretreatments resulted in only slight and inconsistent enhancement of nerve staining. Only the sequential combination of all three pretreatments resulted in consistently better nerve staining.

Barrett, Joyce E.↗

Enhanced Detection of Vibrio Cholerae in Oyster Homogenate Based on Centrifugal Removal of Inhibitory Agents

The disease cholera, caused by Vibrio cholerae, has been associated with consumption of contaminated seafood, including raw oysters. Detection of V. cholerae in foods typically involves blending the oysters, diluting the homogenate in alkaline peptone water (APW), overnight enrichment, and isolation on selective agar. Unfortunately, the oyster homogenate must be diluted to large volumes because lower dilutions inhibit the growth of V. cholerae. The goals of this study were to develop an alternative to large dilutions and to evaluate the basis for the inhibition observed in lower dilutions of oyster homogenates. Centrifugation of oyster homogenates at 10,000 x g for 15 min, followed by enrichment of the resulting pellet in APW, was found to eliminate the inhibition of V. cholerae growth. Inhibition appears not to be due to competing microflora but to a component(s) released when V. cholerae grows in the presence of oyster homogenate. The inhibitory component(s) kills the V. cholerae after the cell concentration reaches > 10(exp 8) cells/mL, rather than initially preventing their growth. The pH also declines from 8.0 to 5.5 during this period; however, the pH decline by itself appears not to cause V. cholerae death. Seven strains of V. cholerae (01 and non-01) and two strains of V. vulnificus were susceptible to the inhibitory agent(s). However, other Vibrio and non-Vibrio species tested were not inhibited by the oyster homogenates. Based on digestion of oyster homogenates with pronase, trypsin and lipase, the inhibitory reaction involves a protein(s). In a preliminary trial with oyster homogenate seeded with 1 cfu/g of V. cholerae, the modified centrifugation technique detected a slightly higher percentage of samples at a 1:10 dilution than the standard FDA Bacteriological Analytical Method (BAM) detected in uncentrifuged oyster homogenate at a 1:100 dilution. V. cholerae in seeded samples could also be detected more frequently by the modified centrifugation method than by PCR at a 1:10 dilution.

Alexander, Donita↗

Genetic Regulation of Charged Particle Mutagenesis in Human Cells

Our studies use a series of syngeneic, and where possible, isogenic human B-lymphoblastoid cell lines to assess the genetic factors that modulate susceptibility apoptosis and their impact on the mutagenic risks of low fluence exposures to 1 GeV Fe ions and 55 MeV protons. These ions are representative of the types of charged particle radiation that are of particular significance for human health in the space radiation environment. The model system employs cell lines derived from the male donor WIL-2. These cells have a single X chromosome and they are hemizygous for one mutation marker, hypoxanthine phosphoribosyltransferase (HPRT). TK6 and WTK1 cells were each derived from descendants of WIL-2 and were each selected as heterozygotes for a second mutation marker, the thymidine kinase (TK) gene located on chromosome 17q. The HPRT and TK loci can detect many different types of mutations, from single basepair substitutions up to large scale loss of heterozygosity (LOH). The single expressing copy of TK in the TK6 and WTKI cell lines is found on the same copy of chromosome 17, and this allele can be identified by a restriction fragment length polymorphism (RFLP) identified when high molecular weight DNA is digested by the SacI restriction endonuclease and hybridized against the cDNA probe for TK. A large series of polymorphic linked markers has been identified that span more than 60 cM of DNA (approx. 60 megabasepairs) and distinguish the copy of chromosome 17 bearing the initially active TK allele from the copy of chromosome 17 bearing the silent TK allele in both TK6 and WTKI cells. TK6 cells express normal p53 protein while WTKI cells express homozygous mutant p53. Expression of mutant p53 can increase susceptibility to x-ray-induced mutations. It's been suggested that the increased mutagenesis in p53 mutant cells might be due to reduced apoptosis.

Kronenberg, Amy↗

Inflight Medical Events in the Shuttle Program

Since the first launch of the Space Shuttle in 1981, the astronauts and their flight surgeons have dealt with a variety of inflight medical issues. A review will be provided of these issues as well as medications used in the treatment of these medical problems. Detailed medical debriefs are conducted by the flight ,surgeon with the individual crewmembers three days after landing. These debriefs were review for Shuttle flights from 1988 through 1999 to determine the frequency of inflight medical events. Medical events were grouped by ICD category and the frequency of medical events within those categories were reviewed. The ICD category of Symptoms, Signs and Ill-defined Conditions had the most medical events. Facial fullness and headache were the most common complaints within this category. The ICD category of Respiratory System had the next most common medical events with sinus congestion being the most common complaint. This was followed by Digestive System complaints and Nervous System/Sense Organ complaints. A variety of inflight medical events have occurred throughout the Shuttle program. Fortunately, the majority of these problems have been minor and have been well within the capability of the medical equipment flown and the skills of the Crew Medical Officers. Medical ,problems/procedures that are routine on the ground often present unique problems in the space flight environment. It is important that the flight surgeon understand the common medical problems encountered.

Baisden, Denise L.↗

Expression of the Acyl-Coenzyme A: Cholesterol Acyltransferase GFP Fusion Protein in Sf21 Insect Cells

The enzyme acyl-coenzyme A:cholesterol acyltransferase (ACAT) is an important contributor to the pathological expression of plaque leading to artherosclerosis n a major health problem. Adequate knowledge of the structure of this protein will enable pharmaceutical companies to design drugs specific to the enzyme. ACAT is a membrane protein located in the endoplasmic reticulum.t The protein has never been purified to homogeneity.T.Y. Chang's laboratory at Dartmouth College provided a 4-kb cDNA clone (K1) coding for a structural gene of the protein. We have modified the gene sequence and inserted the cDNA into the BioGreen His Baculovirus transfer vector. This was successfully expressed in Sf2l insect cells as a GFP-labeled ACAT protein. The advantage to this ACAT-GFP fusion protein (abbreviated GCAT) is that one can easily monitor its expression as a function of GFP excitation at 395 nm and emission at 509 nm. Moreover, the fusion protein GCAT can be detected on Western blots with the use of commercially available GFP antibodies. Antibodies against ACAT are not readily available. The presence of the 6xHis tag in the transfer vector facilitates purification of the recombinant protein since 6xHis fusion proteins bind with high affinity to Ni-NTA agarose. Obtaining highly pure protein in large quantities is essential for subsequent crystallization. The purified GCAT fusion protein can readily be cleaved into distinct GFP and ACAT proteins in the presence of thrombin. Thrombin digests the 6xHis tag linking the two protein sequences. Preliminary experiments have indicated that both GCAT and ACAT are expressed as functional proteins. The ultimate aim is to obtain large quantities of the ACAT protein in pure and functional form appropriate for protein crystal growth. Determining protein structure is the key to the design and development of effective drugs. X-ray analysis requires large homogeneous crystals that are difficult to obtain in the gravity environment of earth. Protein crystals grown in microgravity are often larger and have fewer defects than those grown on earth. The analysis of higher quality space-grown crystals will assist in structure-based drug design. We have successfully grown GCAT-infected Sf21 cells in both adhesion and suspension cultures. Expression levels of GCAT in cell lines such as Sf9 and High Five appear to be reduced. We intend to replicate GCAT expression in all three cell lines using the NASA rotating wall bioreactor which effectively duplicates a microgravity environment. The bioreactor itself could be launched to study the expression of the GFP and GCAT proteins in the actual microgravity environment achieved in orbit.

Mahtani, H. K.↗

Processing and Testing of Thermoplastic Composite Cylindrical Shells Fabricated by Automated Fiber Placement

Two 61-cm-diameter eight-ply quasi-isotropic IM7/PEEK cylindrical shells were fabricated by automated fiber placement the NASA Langley Research Center using only infrared radiant heat to preheat the substrate and incoming composite uni-tape. The shells were characterized by ultrasonic c-scans for overall consolidation quality, and by optical microscopy and acid digestion for void content. Compression tests were also performed. Although the material used in the study was of generally poor quality due to numerous splits and dry fiber regions, the process was able to achieve a net reduction in void content in the as-placed component. Microscopy of the composite shells revealed well-consolidated, void-free interfaces. The two cylinders were then tested in uni-axial compression in a 1334 kN-capacity hydraulic test machine until buckling occurred. A geometrically nonlinear finite element analysis was conducted, and the differences between the predicted and measured values were 18.0 and 25.8%, respectively. Inclusion of measured imperfections of the cylinder into the analysis is expected to reduce these differences.

Hulcher, Anthony Bruce↗

Heat Stress Monitor

The heavy, cumbersome body protection suits worn by members of hazardous materials response teams cause marked elevation of body temperatures, which can reduce effectiveness and lead to heat stress and injury. The CorTemp System, marketed by Human Technologies, Inc., provides the basis for a body temperature monitoring alarm system. Encased in a three-quarter-inch ingestible capsule, the system includes a mini-thermometer, miniature telemetry system, a microbattery and temperature sensor. It makes its way through the digestive system, continuously monitoring temperature. Findings are sent to the recorder by telemetry, and then displayed and stored for transfer to a computer.

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Oil Spill Cleanup

Petroleum Remediation Product (PRP) is a new way of cleaning up oil spills. It consists of thousands of microcapsules, tiny balls of beeswax with hollow centers, containing live microorganisms and nutrients to sustain them. As oil flows through the microcapsule's shell, it is consumed and digested by the microorganisms. Pressure buildup causes the PRP to explode and the enzymes, carbon dioxide and water are released into the BioBoom used in conjunction with PRP, preventing contaminated water from spreading. The system incorporates technology originally developed at the Jet Propulsion Laboratory and Marshall Space Flight Center.

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Sewage Treatment

Stennis Space Center's aquaculture research program has led to an attractive wastewater treatment for private homes. The system consists of a septic tank or tanks for initial sewage processing and a natural secondary treatment facility for further processing of septic tanks' effluent, consisting of a narrow trench, which contains marsh plants and rocks, providing a place for microorganisms. Plants and microorganisms absorb and digest, thus cleansing partially processed wastewater. No odors are evident and cleaned effluent may be discharged into streams or drainage canals. The system is useful in rural areas, costs about $1,900, and requires less maintenance than mechanical systems.

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Cleaner Air for Home and Office

Increased insulation has led to higher concentrations in homes and offices of toxic chemicals caused by emissions from synthetic building components. Recent NASA research regarding future interplanetary manned spacecraft has shown that certain plants can absorb gasses, reducing indoor air pollution. After working with water purifying water hyacinths, Dr. B. C. Wolverton at NSTL developed a carbon/plant filter system to remove chemicals, smoke, etc. Two companies have commercialized the system. Bio-safe provides plants, a bed of activated carbon and an air pump installed near the plant's roots. Pollutants are trapped by the charcoal and either digested by the roots or broken down by microorganisms. Purified air is then directed back into the room. Applied Indoor Resource Company markets Bio-Pure, which includes plants on a layer of patented soil medium with activated carbon. Legumes and mosses filter the air; a blower moves air through the filtering system for cleansing by microorganisms. Research at NSTL continues, and the system may eventually be enlarged.

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