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Description of studies of the kinetics of heat inactivation of phosphoglycerate kinase in the soluble fraction from Hydrogenomonas facilis, its extensive purification, and inhibition by adenosine monophosphate (AMP). No evidence was found for an enzyme which catalyzes adenosine-triphosphate-dependent conversion of 3-phosphoglycerate to 1,3-diphosphoglycerate, AMP, and phosphate.
The response to UV light of several solid proteins and model compounds has been studied in vacuum and at low temperature, using electron paramagnetic resonance techniques. The results indicate that the details of amino acid composition and sequence, and the tertiary structure of a protein are important in determining both the rate of, and the mechanism for, the production of free radicals, and in determining the conditions under which sulfur-type radicals can be produced. The results presented are related to enzyme inactivation and to the UV stability of proteins generally.
Aqueous formaldehyde is shown to exert both sporostatic and sporocidal effects on Bacillus subtilis spores. The sporostatic effect is a result of the reversible inhibition of spore germination occasioned by aqueous formaldehyde; the sporocidal effect is due to the temperature-dependent inactivation of these spores in aqueous formaldehyde. The physicochemical state of formaldehyde in solution provides a framework with which to interpret both the sporostatic and sporocidal properties of aqueous formaldehyde.
Saturethane solid propellant was used in all tests. The spore inoculum was evenly distributed in the propellant. Samples weighing approximately 5 g were aseptically removed, placed into curing ovens, and exposed to cure temperatures. Initial tests were conducted at 82 and 93 C. Analysis of the obtained data indicated that the survivor curves were not linear. Exposure of the inoculated propellant samples to 93 C reduced the initial population to less than 0.01% in about 20 hours. At 82 C, approximately 168 hours were required for a similar reduction. Tests involving curing temperatures of 105 and 115 C were also conducted. It is pointed out that changes in the mechanism of spore inactivation due to chemical and physical changes in the propellant could account for the nonlinear survivor curves obtained.
The viability and dry heat resistance of indigenous microflora associated with small soil particles were investigated. An aluminum boat TDT CUP-TSA solid media system was developed for the analyses; a complete description of the technique is included. Data cited here were obtained using analyses of individual soil particles. Detailed particle viability profiles for dry heat effects were determined for Kennedy Space Center soil. At 110 C at least some particles retained viability through a heating period of between 8 and 16 hours. Single particles heated at 125 C for 80 minutes or longer did not show evidence of viability under test conditions. Preliminary aerobic, mesophilic plate counts of the 74-88 micron m soil fraction yielded mean values of 16.2 organisms per dark particle and 2.6 organisms per light particle. Heat treatment of particles in a dry atmosphere did not appear to increase the rate of inactivation for in situ soil particle microflora.
Experimental evidence indicates that the radiation dose required to reduce a surviving population to a certain fraction of its original population is lower for vertebrate cells than for viruses. On the other hand, the number of ionizations per cell required to inactivate that cell is greater for vertebrate cells than for viruses. The apparent conflict between these two findings is investigated. It is found that the apparent contradiction is probably a result of the fractional power dependence of the radiation-dose value on the nucleic acid weight.
The likelihood of biologically contaminating a planet by an atmospheric probe has a low probability of occurring if the probe is kept biologically clean during terrestrial operations and if the structure remains in tact until the planets life zone is completely penetrated. High standards of cleanliness, monitoring and estimating for remedial actions must be maintained in a probe program. It is not a foregone conclusion, however, that heat sterilization needs to be employed. The use of several techniques having a good potential for lower probe costs are available and appear adequate to render a probe sterile within acceptable bounds. The techniques considered to be satisfactory for minimizing microbial load include: (1) combined heat (at 95-105 C) and gamma radiation; (2) short term heating at 105 + or - 5 C to inactivate all vegetative microbes; (3) irradiation routinely by ultraviolet light; (4) wiping by a bactericidal agent with or without a penetrant; and (5) cleanliness alone.
Based on information concerning the interaction of salts and macromolecules the literature of the enzymes of halophilic bacteria and their constituents is examined. Although in halophilic systems the salt requirement of enzyme activity is variable the enzymes investigated show a time-dependent inactivation at lower salt concentrations especially in the absence of salt. The studies described show that in some halophilic systems the effect of salt may be restricted to a small region on the protein molecule. The concept of the hydrophobic bond to consider certain solvent-dependent phenomena is introduced. It is shown that some halophilic enzymes are unable to maintain their structure without the involvement of hydrophobic interactions that are usually not supported by water. A table lists indices of hydrophobicity and polarity for various halophilic and nonhalophilic proteins.
Responses of normothermic and hibernating marmots to manipulations of the preoptic-hypothalamic temperature (T-PO) were experimentally investigated. An exponential increase in open-loop gain (OLG) occurred with decreases in temperature; it is concluded that this response can be explained by recruitment of cold-sensitive neurons brought about by low-temperature inactivation of inhibitory neurons. Marmots not only seek out the hibernating state, but also utilize all the thermoregulatory means they possess to remain in it for a given period of time.
Spores of Bacillus subtilis HA 101 and the DNA polymerase I-defective mutant HA 101 (59)F were exposed to selected wavelengths of solar ultraviolet light and space vacuum during the return of Apollo 16. In addition, coliphage T-7 suspensions were exposed to solar ultraviolet radiation as part of the Microbial Response to Space Environment Experiment. Optical filters were employed to provide different energy levels at wavelengths 254 nm and 280 nm. Dose-response curves for lethal and mutagenic effects were compared with ground-based data. A close parallel was observed between the results of solar radiation and ground tests with spores of the two strains. However, significantly greater inactivation of T-7 bacteriophage was observed after exposure to solar ultraviolet radiation.
System between the Titan/Centaur launch vehicle and its aerodynamic shroud is described. The system provides a precise spring constant and is capable of being inactivated during flight. Design requirements, design details, and the test program are discussed. The conventional English system of units was used during this development program for all principal measurements and calculations.
This space bioprocessing contract effort was comprised of four general objectives. These were: (1) the evaluation of current separation processes, (2) the identification of problems relevant to the separation of important biologicals, (3) the identification of ground-based assay methods needed for pre- and postflight analysis of space bioprocessing separation technology; and (4) the establishment of methods to determine the efficiency of space bioprocessing separation procedures. Immunology was deemed advantageous to study the diversity of cells and cell products involved and the extensive interest being given to their separation. Upon recognition of a cellular or molecular agent as foreign to the body, the immune system becomes activated to produce cells whose function is to destroy that agent and cell products whose function is to inactivate the agent and assist in its destruction. Long after the agent is removed from the body, some cells remain in a state of readiness to continue these destructive actions specifically against that agent should further exposure to it occur. This is the basis of acquired immunity to disease.
Evaluations on critical components of the mobile biological isolation system were performed. High efficiency particulate air filter efficiency and suit integrity were found to withstand repeated ethylene oxide (ETO) sterilizations. The minimum ETO sterilization time required to inactivate all contaminant organisms was established at four hours. Two days of aerating at 120 F was found to dissipate all harmful ETO residuals from the suit. Donning and doffing procedures were clarified and written specifically for isolation rooms.
Active serine accumulation in cell envelope vesicles from Halobacterium halobium proceeds by co-transport with Na(+) and can be induced by either transmembrane electrical potential or transmembrane Na(+) concentration difference. It was shown earlier that in the former case the initial transport rate is a fourth-power function of the magnitude of the electrochemical potential difference of sodium ions, and in the latter, a second-power function. A possible interpretation of this finding is cooperativity of sodium-transporting sites in the transport carrier. When both kinds of driving force are imposed simultaneously on the vesicles, fourth-power dependence on the total potential difference of sodium ions is obtained, suggesting that the transport carrier is regulated by the electrical potential. Heat treatment of the vesicles at 48 C partially inactivates transport and abolishes this effect of the electrical potential.
A method and apparatus for removing porphyrins from a fluid sample which are unrelated to the number of bacteria present in the sample and prior to combining the sample with luminol reagent to produce a light reaction is disclosed. The method involves a pre-incubation of the sample with a dilute concentration of hydrogen peroxide which inactivates the interfering soluble porphyrins. Further, by delaying taking a light measurement for a predetermined time period after combining the hydrogen peroxide-treated water sample with a luminol reagent, the luminescence produced by the reaction of the luminol reagent with ions present in the solution, being short lived, will have died out so that only porphyrins within the bacteria which have been released by rupturing the cells with the sodium hydroxide in the luminol reagent, will be measured. The measurement thus obtained can then be related to the concentration of live and dead bacteria in the fluid sample.
A 50.8-cm-tip-diameter turbine equipped with a rotor tip shroud of hexagonal cell (or honeycomb) cross section has been tested in warm air (416 K) for a range of shroud coolant to primary flow rates. Test results were also obtained for the same turbine operated with a solid shroud for comparison. The results showed that the combined effect of the honeycomb shroud and the coolant flow was to cause a reduction of 2.8 points in efficiency at design speed, pressure ratio, and coolant flow rate. With the coolant system inactivated, the honeycomb shroud caused a decrease in efficiency of 2.3 points. These results and those obtained from a small reference turbine indicate that the dominant factor governing honeycomb tip shroud loss is the ratio of honeycomb depth to blade span. The loss results of the two shrouds could be correlated on this basis. The same honeycomb and coolant effects are expected to occur for the hot (2200 K) version of this turbine.
The mechanism of the prostacyclin-linked hyperthermia is studied in rabbits. Results show that intracerebroventricular administration of prostacyclin (PGI2) induces dose-related hyperthermia at room temperature (21 C), as well as at low (4 C) and high (30 C) ambient temperatures. It is found that this PGI2-induced hyperthermia is not mediated by its stable metabolite 6-keto prostaglandin F-1(alpha). Only one of the three anion transport systems, the liver transport system, appears to be important to the central inactivation of pyrogen, prostaglandin E2, and PGI2. Phenoxybenzamine and pimozide have no thermolytic effect on PGI2-induced hyperthermia, while PGI2 still induces hyperthermia after norepinephrine (NE) and dopamine levels are depleted by 6-hydroxydopamine. Indomethacin and SC-19220 (a PG antagonist) do not antagonize PGI2 induced hyperthermia, while theophylline does not accentuate the PGI2-induced hyperthermia. However, the hyperthermic response to PGI2 is attenuated by central administration of the protein synthesis inhibitor, anisomycin. It is concluded that PGI2-induced hyperthermia is not induced by NE, dopamine, or cyclic AMP, but rather that a protein mediator is implicated in the induction of fever by PG12.
The intracerebroventricular administration of prototype nonpeptide opioid receptor (mu, kappa, and sigma) agonists, morphine, ketocyclazocine, and N-allyl normetazocine and an agonist at both kappa and sigma receptors, pentazocine, was found to induce hyperthermia in guinea pigs. The similar administration of peptide opioids like beta endorphin, methionine endkephalin, leucine endkephaline, and several of their synthetic analogues was also found to cause hyperthermia. Only the liver-like transport system of the three anion transport systems (iodide, hippurate, and liver-like) present in the choroid plexus was determined to be important to the central inactivation of beta-endorphin and two synthetic analogues. Prostaglandins and norepinephrine (NE) as well as cAMP were not involved in peptide and nonpeptide opioid-induced hyperthermia. Naloxone-sensitive receptors were found to be involved in the induction of hyperthermia by morphine and beta-endorphin, while hyperthermic responses to ketocyclazocine, N-allyl normetazocine, pentazocine, Met-enkephalin, Leu-enkephalin, and two of the synthetic analogues were not antagonized by nalozone. The lack of antagonism of naloxone on pyrogen, arachidonic acid, PGE2, dibutyryl cAMP, and NE-induced hyperthermia shows that endogenous opioid peptides are not likely to be central mediators of the hyperthermia induced by these agents.