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At least 91 records · Page 5

Testing First-Order Logic Axioms in AutoCert

AutoCert [2] is a formal verification tool for machine generated code in safety critical domains, such as aerospace control code generated from MathWorks Real-Time Workshop. AutoCert uses Automated Theorem Provers (ATPs) [5] based on First-Order Logic (FOL) to formally verify safety and functional correctness properties of the code. These ATPs try to build proofs based on user provided domain-specific axioms, which can be arbitrary First-Order Formulas (FOFs). These axioms are the most crucial part of the trusted base, since proofs can be submitted to a proof checker removing the need to trust the prover and AutoCert itself plays the part of checking the code generator. However, formulating axioms correctly (i.e. precisely as the user had really intended) is non-trivial in practice. The challenge of axiomatization arise from several dimensions. First, the domain knowledge has its own complexity. AutoCert has been used to verify mathematical requirements on navigation software that carries out various geometric coordinate transformations involving matrices and quaternions. Axiomatic theories for such constructs are complex enough that mistakes are not uncommon. Second, adjusting axioms for ATPs can add even more complexity. The axioms frequently need to be modified in order to have them in a form suitable for use with ATPs. Such modifications tend to obscure the axioms further. Thirdly, speculating validity of the axioms from the output of existing ATPs is very hard since theorem provers typically do not give any examples or counterexamples.

Ahn, Ki Yung↗

Alternate Protocol for Detecting Biological Contamination on Sensitive Hardware

The purpose of this project is to develop a sterile water based rapid bioburden test. Contamination engineers use two tests to assess the level of biological contamination on hardware: the rapid five minute bioburden test, which is a molecular screening for Adenosine triphosphate (ATP), a molecule found in all cells on the hardware, and a slower colony growth test, which is used to give a more accurate representation of the amount of microbes on the hardware. However, the rapid bioburden test has limited application because it leaves a residue that can be detrimental to sensitive hardware. This can cause project delays while waiting for the results from the three day colony growth test. We address this problem by adapting the commercial germicide based ATP system to a sterile water based system. The test works by reacting ATP with D-Luciferin and Luciferase protein to yield light. The light is then detected by a luminometer that outputs a Relative Light Unit (RLU) amount depending on how much ATP is present. To analyze the effectiveness of the new test, we developed a correlation between amounts of ATP and the RLU produced using the germicide based system. From these experiments, we've generated a consistent relationship between the two in the form of a power curve. From there, we developed a correlation curve between the amount of colonies and the RLU they produced. Initial tests of the new protocol have shown that the water based system isn't as sensitive as the germicide based test.

planetary protection↗

Method of detecting and counting bacteria in body fluids

A novel method is reported for determining bacterial levels in urine samples, which method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of non-bacterial ATP. After the removal of non-bacterial ATP, the bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay using an enzyme obtained from the firefly.

Chappelle, E. W.↗

Dicyclohexylcarbodiimide-sensitive ATPase in Halobacterium saccharovorum

Membranes from Halobacterium saccharovorum contained a cryptic ATPase which required Mg(2+) or Mn(2+) and was activated by Triton X-100. The optimal pH for ATP hydrolysis was 9-10. ATP or GTP were hydrolyzed at the same rate while ITP, CTP, and UTP were hydrolyzed at about half that rate. The products of ATP hydrolysis were ADP and phosphate. The ATPase required high concentrations (3.5 M) of NaCl for maximum activity. ADP was a competitive inhibitor of the activity, with an apparent Ki of 50 micro-M. Dicyclohexylcarbodiimide (DCCD) inhibited ATP hydrolysis. The inhibition was marginal at the optimum pH of the enzyme. When the ATPase was preincubated with DCCD at varying pH values, but assayed at the optimal pH for activity, DCCD inhibition was observed to increase with increasing acidity of the preincubation medium. DCCD inhibition was also dependent on time of preincubation, and protein and DCCD concentrations. When preincubated at pH 6.0 for 4 h at a protein:DCCD ratio of 40 (w/w), ATPase activity was inhibited 90 percent.

Kristjansson, H.↗

Computational Fluid Dynamics (CFD) Analyses in Support of Space Shuttle Main Engine (SSME) Heat Exchanger (HX) Vane Cracking Investigation

Integration issues involved with installing the alternate turbopump (ATP) High Pressure Oxygen Turbopump (HPOTP) into the SSME have raised questions regarding the flow in the HPOTP turnaround duct (TAD). Steady-state Navier-Stokes CFD analyses have been performed by NASA and Pratt & Whitney (P&W) to address these questions. The analyses have consisted of two-dimensional axisymmetric calculations done at Marshall Space Flight Center and three-dimensional calculations performed at P&W. These analyses have identified flowfield differences between the baseline ATP and the Rocketdyne configurations. The results show that the baseline ATP configuration represents a more severe environment to the inner HX guide vane. This vane has limited life when tested in conjunction with the ATP but infinite life when tested with the current SSME HPOTP. The CFD results have helped interpret test results and have been used to assess proposed redesigns. This paper includes details of the axisymmetric model, its results, and its contribution towards resolving the problem.

Garcia, Roberto↗

Microbial assessment of cabin air quality on commercial airliners

The microbial burdens of 69 cabin air samples collected from commercial airliners were assessed via conventional culture-dependent, and molecular-based microbial enumeration assays. Cabin air samples from each of four separate flights aboard two different carriers were collected via air-impingement. Microbial enumeration techniques targeting DNA, ATP, and endotoxin were employed to estimate total microbial burden. The total viable microbial population ranged from 0 to 3.6 x10 4 cells per 100 liters of air, as assessed by the ATP-assay. When these same samples were plated on R2A minimal medium, anywhere from 2% to 80% of these viable populations were cultivable. Five of the 29 samples examined exhibited higher cultivable counts than ATP derived viable counts, perhaps a consequence of the dormant nature (and thus lower concentration of intracellular ATP) of cells inhabiting these air cabin samples. Ribosomal RNA gene sequence analysis showed these samples to consist of a moderately diverse group of bacteria, including human pathogens. Enumeration of ribosomal genes via quantitative-PCR indicated that population densities ranged from 5 x 10 1 ' to IO 7 cells per 100 liters of air. Each of the aforementioned strategies for assessing overall microbial burden has its strengths and weaknesses; this publication serves as a testament to the power of their use in concert.

microbe↗

Improved methods for counting bacteria in physiological fluids

Bacterial population detection is based on detection of adenosine triphosphate (ATP), chemical present in all living matter. Amount of ATP in sample, after chemically removing all nonbacterial ATP, is directly related to bacterial population. Sensitivity is improved by concentration step; specificity is improved by lowering pH of solution.

Picciolo, G. L.↗

Bioluminescence for determining energy state of plants

Bioluminescence produced by the luciferin-luciferase system is a very sensitive assay for ATP content in extracts of plant materials. The ATP test for seed and pollen viability and vigor is presented, along with prediction of high growth potential and productivity in new crosses and selections of breeding materials. ATP as an indicator for environmental quality, stresses, and metabolic regulation is also considered.

Ching, T. M.↗

Method of detecting and counting bacteria

An improved method is provided for determining bacterial levels, especially in samples of aqueous physiological fluids. The method depends on the quantitative determination of bacterial adenosine triphosphate (ATP) in the presence of nonbacterial ATP. The bacterial ATP is released by cell rupture and is measured by an enzymatic bioluminescent assay. A concentration technique is included to make the method more sensitive. It is particularly useful where the fluid to be measured contains an unknown or low bacteria count.

Picciolo, G. L.↗

Determination of antimicrobial susceptibilities on infected urines without isolation

A method is described for the quick determination of the susceptibilities of various unidentified bacteria contained in an aqueous physiological fluid sample, particularly urine, to one or more antibiotics. A bacterial adenosine triphosphate (ATP) assay is carried out after the elimination of non-bacterial ATP to determine whether an infection exists. If an infection does exist, a portion of the sample is further processed, including subjecting parts of the portion to one or more antibiotics. Growth of the bacteria in the parts are determined, again by an ATP assay, to determine whether the unidentified bacteria in the sample are susceptible to the antibiotic or antibiotics under test.

Picciolo, G. L.↗

Effect of immobilization stress on the level of macroergic phosphates in the blood of rats

The effect of immobilization for 1, 2, and 24 hours, and of daily (for 2 hours for 7 days) immobilization on the blood nucleotide (ATP, ADP, AMP) level was studied on 79 male rats. A progressive reduction of the ATP content was most pronounced in immobilization for 24 hours. This was accompanied by an increase of lymphopenia and eosinopenia. A fall of the relative weight of the thymus and a weight gain of the adrenal glands was observed along with a reduction of ascorbic acid concentration in both of them. In case of daily immobilization for 2 hours, the ATP and ADP content on the 1st and 2nd day was below the normal level, and then showed a gradual increase, with complete normalization in 6-7 days, except for inorganic phosphorus the level of which remained lower than normal.

Pudov, V. I.↗

A quinone-assisted photoformation of energy-rich chemical bonds

In a study of biochemical means of solar energy conversion, ADP and inorganic phosphates were converted to ATP by white light in the nonaqueous solvent dimethylformamide in the presence of tetrachloro-p-quinone or ubiquinone. Conversion of ADP to ATP has been accomplished in aqueous suspension by the use of cell-like structures aggregated from poly(aspartic acid, glutamic acid, tyrosine). This is believed to occur through the formation of dopaquinone in the peptide structure during illumination. The way in which the quantitative yield of ATP has been influenced by pH and by added substances, such as FeCl2, was studied.

Fox, S. W.↗

The effects of cutting or of stretching skeletal muscle in vitro on the rates of protein synthesis and degradation

Skeletal muscle preparations using cut muscle fibers have often been used in studies of protein metabolism. The present paper reports an investigation of the effect of muscle cutting or stretching in vitro on the rates of protein synthesis and/or degradation. Protein synthesis and content, and ATP and phosphocreatine levels were monitored in soleus and extensor digitorum longus muscles from the rat with various extents of muscle fiber cuts and following stretching to about 120% the resting length. Rates of protein synthesis are found to be significantly lower and protein degradation higher in the cut muscles than in uncut controls, while ATP and phosphocreatine concentrations decreased. Stretched intact muscles, on the other hand, are observed to have higher concentrations of high-energy phosphates than unstretched muscles, while rates of protein degradation were not affected. Results thus demonstrate that the cutting of skeletal muscle fibers alters many aspects of muscle metabolism, and that moderate decreases in ATP concentration do not alter rates of protein concentration in intact muscles in vitro.

Seider, M. J.↗

Models of glycolysis: Glyceraldehyde as a source of energy and monomers for prebiotic condensation reactions

All organisms require energy in a chemical form for maintenance and growth. In contemporary life this chemical energy is obtained by the synthesis of the phosphoanhydride bonds of ATP. Among the biological processes that yield ATP, fermentation is generally considered primitive, because it operates under anaerobic conditions by substrate-level phosphorylation which does not require compartmentation by membranes. Fermentation by the glycolytic pathway, which is found in almost every living cell, is an especially attractive energy source for primitive life. Glycolysis not only produces useful chemical energy (ATP), but intermediates of this pathway are also involved in amino acid synthesis and photosynthetic carbon-fixation. It is believed that energy and substrates needed for the origin of life were provided by nonenzymatic chemical reactions that resemble the enzyme-mediated reactions of glycolysis. These nonenzymatic reactions would have provided a starting point for the evolutionary development of glycolysis.

Weber, A. L.↗

Annoyance response to simulated advanced turboprop aircraft interior noise containing tonal beats

A study is done to investigate the effects on subjective annoyance of simulated advanced turboprop (ATP) interior noise environments containing tonal beats. The simulated environments consisted of low-frequency tones superimposed on a turbulent-boundary-layer noise spectrum. The variables used in the study included propeller tone frequency (100 to 250 Hz), propeller tone levels (84 to 105 dB), and tonal beat frequency (0 to 1.0 Hz). Results indicated that propeller tones within the simulated ATP environment resulted in increased annoyance response that was fully predictable in terms of the increase in overall sound pressure level due to the tones. Implications for ATP aircraft include the following: (1) the interior noise environment with propeller tones is more annoying than an environment without tones if the tone is present at a level sufficient to increase the overall sound pressure level; (2) the increased annoyance due to the fundamental propeller tone frequency without harmonics is predictable from the overall sound pressure level; and (3) no additional noise penalty due to the perception of single discrete-frequency tones and/or beats was observed.

Leatherwood, Jack D.↗

NASA/industry advanced turboprop technology program

Experimental and analytical effort shows that use of advanced turboprop (propfan) propulsion instead of conventional turbofans in the older narrow-body airline fleet could reduce fuel consumption for this type of aircraft by up to 50 percent. The NASA Advanced Turboprop (ATP) program was formulated to address the key technologies required for these thin, swept-blade propeller concepts. A NASA, industry, and university team was assembled to develop and validate applicable design codes and prove by ground and flight test the viability of these propeller concepts. Some of the history of the ATP project, an overview of some of the issues, and a summary of the technology developed to make advanced propellers viable in the high-subsonic cruise speed application are presented. The ATP program was awarded the prestigious Robert J. Collier Trophy for the greatest achievement in aeronautics and astronautics in America in 1987.

Ziemianski, Joseph A.↗

NASA/industry advanced turboprop technology program

Experimental and analytical effort shows that use of advanced turboprop (propfan) propulsion instead of conventional turbofans in the older narrow-body airline fleet could reduce fuel consumption for this type of aircraft by up to 50 percent. The NASA Advanced Turboprop (ATP) program was formulated to address the key technologies required for these thin, swept-blade propeller concepts. A NASA, industry, and university team was assembled to develop and validate applicable design codes and prove by ground and flight test the viability of these propeller concepts. Some of the history of the ATP Project, an overview of some of the issues, and a summary of the technology developed to make advanced propellers viable in the high-subsonic cruise speed application are presented. The ATP program was awarded the prestigious Robert J. Collier Trophy for the greatest achievement in aeronautics and astronautics in America in 1987.

Ziemianski, Joseph A.↗

Energy from Redox Disproportionation of Sugar Carbon Drives Biotic and Abiotic Synthesis

To identify the energy source that drives the biosynthesis of amino acids, lipids, and nucleotides from glucose, we calculated the free energy change due to redox disproportionation of the substrate carbon of: (1) 26-carbon fermentation reactions and (2) the biosynthesis of amino acids and lipids of E. coli from glucose. The free energy (cal/mmol of carbon) of these reactions was plotted as a function of the degree of redox disproportionation of carbon (disproportionative electron transfers (mmol)/mmol of carbon). The zero intercept and proportionality between energy yield and degree of redox disproportionation exhibited by this plot demonstrate that redox disproportionation is the principal energy source of these redox reactions (slope of linear fit = -10.4 cal/mmol of disproportionative electron transfers). The energy and disproportionation values of E. coli amino acid and lipid biosynthesis from glucose lie near this linear curve fit with redox disproportionation accounting for 84% and 96% (and ATP only 6% and 1 %) of the total energy of amino acid and lipid biosynthesis, respectively. These observations establish that redox disproportionation of carbon, and not ATP, is the primary energy source driving amino acid and lipid biosynthesis from glucose. In contrast, we found that nucteotide biosynthesis involves very little redox disproportionation, and consequently depends almost entirely on ATP for energy. The function of sugar redox disproportionation as the major source of free energy for the biosynthesis of amino acids and lipids suggests that sugar disproportionation played a central role in the origin of metabolism, and probably the origin of life.

Weber, Arthur L.↗