Biomedical subjects
Publications and source records attributed to H P Klein.
A major argument in the claim that life had been discovered during the Viking mission to Mars is that the results obtained in the Labeled Release (LR) experiment are analogous to those observed with terrestrial microorganisms. This assertion is critically examined and found to be implausible.
Proposals for continuing the search for extant life on Mars are primarily predicated on the assumption that specialized environmental niches that could support a biota may exist on the planet. Before attempting any critical tests for extant organisms, either in situ or on returned samples, it is imperative to determine whether any such sites actually exist. If, through remote sensing and landed instrumentation, sites of potential biological interest are discovered and characterized, biological tests can then more effectively be planned to elicit the presence of organisms that are adapted to living in these particular environments.
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Of all the other planets in the solar system, Mars remains the most promising for further elucidating concepts about chemical evolution and the origin of life. Strategies were developed to pursue three exobiological objectives for Mars exploration: determining the abundance and distribution of the biogenic elements and organic compounds, detecting evidence of an ancient biota on Mars, and determining whether indigenous organisms exist anywhere on the planet. The three strategies are quite similar and, in fact, share the same sequence of phases. In the first phase, each requires global reconnaissance and remote sensing by orbiters to select sites of interest for detailed in situ analyses. In the second phase, lander missions are conducted to characterize the chemical and physical properties of the selected sites. The third phase involves conducting "critical" experiments at sites whose properties make them particularly attractive for exobiology. These critical experiments would include, for example, identification of organics, detection of fossils, and detection of extant life. The fourth phase is the detailed analysis of samples returned from these sites in Earth-based laboratories to confirm and extend previous discoveries. Finally, in the fifth phase, human exploration is needed to establish the geological settings for the earlier findings or to discover and explore sites that are not accessible to robotic spacecraft.
In looking ahead to possible new attempts to search for extant life on Mars, the history of the Viking biological investigations is reviewed here. Scientific considerations that led to the selection of specific experimental approaches for life detection are discussed, as well as the overall results obtained from that mission. Despite extensive preflight testing of the concepts that were to be used, unanticipated artefacts arose in the actual mission. These almost certainly reflect the fact that, at that time, there were many gaps in our understanding of the physical and chemical characteristics of the Martian environment. After Viking, many of these issues still remain unresolved, and future attempts to search for extant biology should be restrained until adequate new information about potential habitable microenvironments is obtained.
The mechanism for synthesis of monounsaturated fatty acids under aerobic and anaerobic conditions was studied in the facultative anaerobic cyanobacterium, Oscillatoria limnetica. The hexadecenoic acid (C16:1) of aerobically grown O. limnetica was shown to contain both the delta 7 (79%) and delta 9 (21%) isomers, while the octadecenoic (C18:1) acid was entirely the delta 9 acid. Incorporation of [2-14C] acetate into the fatty acids under aerobic conditions resulted in synthesis of the delta 7 and delta 9 C16:1 and the delta 9 C18:1. Synthesis of unsaturated fatty acids in the presence of DCMU required sulfide. Anaerobic incubations in the presence of DCMU and sulfide (less than 0.003% atmospheric oxygen) resulted in a two-fold increase in monounsaturated fatty acids of both delta 7 and delta 9 C16:1 and delta 9 and delta 11 C18:1. The synthesis of these is characteristic of a bacterial-type, anaerobic pathway.
Sample return missions from a comet nucleus and the Mars surface are currently under study in the US, USSR, and by ESA. Guidance on Planetary Protection (PP) issues is needed by mission scientists and engineers for incorporation into various elements of mission design studies. Although COSPAR has promulgated international policy on PP for various classes of solar system exploration missions, the applicability of this policy to sample return missions, in particular, remains vague. In this paper, we propose a set of implementing procedures to maintain the scientific integrity of these samples. We also propose that these same procedures will automatically assure that COSPAR-derived PP guidelines are achieved. The recommendations discussed here are the first step toward development of official COSPAR implementation requirements for sample return missions.
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At onset of chronic rheumatoid arthritis a 36-year-old woman was started on a course of sulphasalazine. During the first four weeks the treatment she developed severe dyspnoea, mild fever, dry cough with chest pain, marked hypoxaemia and severely abnormal restrictive lung functions. Chest x-ray demonstrated diffuse alveolar-interstitial infiltrates. After discontinuing the drug and short-term administration of corticosteroids, blood gases and the chest x-ray reverted to normal within four weeks, but the abnormal lung functions persisted. The course of the illness and published reports on the side effects of sulphasalazine point to the need of carefully watching out for possible side effects during the first three months of treatment with this drug.
The term "Exobiology" was introduced about 25 years ago, at a time when intensive discussions were under way concerning plans for the biological exploration of Mars. The search for life on Mars was to be a critical test of the concept of chemical evolution--not an end in itself. After the Viking mission, when it became apparent that prospects were dim for the discovery of extraterrestrial life within our solar system, many people concluded that this new field of endeavor would soon expire. Quite the contrary, over the past decade, the field had broadened considerably into a multidisciplinary approach to understanding the circumstances that led to the origin of life and the interplay between the evolution of this planet and its biota.
The stability of particulate palmitoyl-CoA desaturase preparations from anaerobically grown yeast cells was increased by exposure to low levels of oxygen. The stabilizing effect of oxygen may be based upon the increased amounts of palmitoleic acid and ergosterol that become available to the cells. These results suggest the evolutionary appearance of this system at a time when atmospheric oxygen was at a low level.
Spacelab is a large versatile laboratory carried in the bay of the Shuttle Orbiter. The first Spacelab mission dedicated entirely to Life Sciences is known as Spacelab 4. It is scheduled for launch in late 1985 and will remain aloft for seven days. This payload consists of 25 tentatively selected investigations combined into a comprehensive integrated exploration of the effects of acute weightlessness on living systems. An emphasis is placed on studying physiological changes that have been previously observed in manned space flight. This payload has complementary designs in the human and animal investigations in order to validate animal models of human physiology in weightlessness. The experimental subjects include humans, squirrel monkeys, laboratory rats, several species of plants, and frog eggs. The primary scientific objectives include study of the acute cephalic fluid shift, cardiovascular adaptation to weightlessness, including postflight reductions in orthostatic tolerance and exercise capacity, and changes in vestibular function, including space motion sickness, associated with weightlessness. Secondary scientific objectives include the study of red cell mass reduction, negative nitrogen balance, altered calcium metabolism, suppressed in vitro lymphocyte reactivity, gravitropism and photropism in plants, and fertilization and early development in frog eggs. The rationale behind this payload, the selection process, and details of the individual investigations are presented in this paper.
The effect of oxygen on squalene epoxidase activity in Saccharomyces cerevisiae was investigated. In cells grown in standing cultures, the epoxidase was localized mainly in the "mitochondrial" fraction. Upon aeration, enzyme activity increased and the newly formed enzyme was associated with the "microsomal" fraction. At 0.03% (vol/vol) oxygen, epoxidase levels doubled, whereas the ergosterol level was only slightly increased. Cycloheximide inhibited the increase in epoxidase under these conditions. An apparent Km for oxygen of 0.38% (vol/vol) was determined from a crude particulate preparation for the epoxidase.
The effect of "weightlessness" on liver metabolism was examined using tissue from rats flown in earth orbit for 18.5 days aboard the Soviet Cosmos 936 biosatellite. Changes in the activities of certain carbohydrate and lipid enzymes were noted. Of the 28 hepatic enzyme activities assayed, two, palmitoyl-CoA desaturase and lactate dehydrogenase, increased, whereas five, glycogen phosphorylase, 6-phosphogluconate dehydrogenase, both acyltransferases which act on alpha-glycerolphosphate and diglycerides, and aconitate hydratase decreased. The remaining enzyme activities measured were unchanged. In addition, increased levels of liver glycogen and palmitoleate were noted which probably resulted from the lowered glycogen phosphorylase and increased palmitoyl-CoA desaturase activities, respectively, in those animals that experienced weightlessness. These changes caused by weightlessness were transient since all of the aforementioned alterations returned to normal values when measured in the livers of other rats which had flown in the biosatellite 25 days after recovery.
Factors related to pathogenesis were studied in a murine model of pulmonary fungal infection with strains of Blastomyces dermatitidis. Strain 26199V was greater than or equal to 10(5)-fold more lethal in pulmonary challenge than strain GA-1. This relationship between the two strains also occurred after intraperitoneal challenge. Serial studies with lung cultures and histology of sacrificed animals after pulmonary challenge with similar doses, and challenge studies in mice of differing maturity, indicated GA-1 is impaired in initial replication in situ, although the two strains grow equally well in the presence of murine substrates in vitro. In vitro studies indicated the two strains were indistinguishable in most studies of biochemical and metabolic markers, and in studies with chemical and physical inhibitors. The exceptions were their morphologic appearance in the yeast phase, ability to hydrolyze esculin, and susceptibility to crystal violet dye and bile. The last finding and previous work suggested possible differences in lipid content; these were confirmed in studies which also included two other strains. Greater phospholipid and palmitic acid content were associated with greater virulence.
A spontaneous mutation occurred in cultures of a virulent strain of Blastomyces dermatitidis during serial passage at 35-37 degrees C. The mutant strain was shown to be 10,000-fold less virulent for mice than the parent strain, and this was independent of the challenge dose, the age of the challenged host, and the route of challenge (although attenuation was greater for pulmonary than for peritoneal challenge). The mutant grows as well as the parent in fungal media or in the presence of murine substrates. Studies of the chronology of pulmonary infection indicate the mutation affects an early event (first week after challenge) in establishment of infection and interaction with host defenses. Scanning electron microscopy showed that the mutant lacks deep irregular surface ridges and grooves present in the parent. It had increased resistance to crystal violet in vitro, increased susceptibility to bile, and could hydrolyze esculin. The latter findings are of interest because they place the mutant in an intermediate position with respect to these properties between the virulent parent and a previously studied avirulent strain. Lipid analyses indicated a marked increase in fatty acids in the mutant. Studies of four B. dermatitidis strains have also associated increasing palmitic acid and phospholipid content with increasing virulence.
We have examined, in the livers of rats carried aboard the Cosmos 936 biosatellite, the activities of about 30 enzymes concerned with carbohydrate and lipid metabolism. In addition to the enzyme studies, the levels of glycogen and of the individual fatty acids in hepatic lipids were determined. Livers from flight and ground control rats at recovery (R0) and 25 days after recovery (R25) were used for these analyses. For all parameters measured, the most meaningful comparisons are those made between flight stationary (FS) and flight centrifuged (FC) animals at RQ. When these two groups of flight rats were compared at R0, statistically significant decreases in the activity levels of glycogen phosphorylase, alpha-glycerol phosphate acyl transferase, diglyceride acyl transferase, aconitase and 6-phosphogluconate dehydrogenase and an increase in the palmitoyl CoA desaturase were noted in the weightless group (FS). The significance of these findings was strengthened by the fact that all enzyme activities showing alterations at R0 returned to normal 25 days postflight. When liver glycogen and total fatty acids of the two sets of flight animals were determined, significant differences that could be attributed to reduced gravity were observed. The weightless group (FS) at R0 contained, on the average, more than twice the amount of glycogen than did the centrifuged controls (FC) and a remarkable shift in the ratio of palmitate to palmitoleate was noted. These metabolic alterations, both in enzyme levels and in hepatic constituents, appear to be characteristic of the weightless condition. Our data seem to justify the conclusion that centrifugation during flight is equivalent to terrestrial gravity.
1. In the present paper, the two acetyl-CoA synthetases (acetate:Coenzyme A ligase (AMP-forming), EC 6.2.1.1) elaborated under aerobic or nonaerobic conditions are further differentiated by an immunological approach. 2. The subunit of the aerobic isozyme was prepared and found to be homogeneous by disc gel electrophoresis in the presence of sodium dodecyl sulfate (SDS) and by ultracentrifugal studies. An s20,w of 3.6 and an apparent molecular weight of 80,500 +/- 500 were calculated for this subunit. 3. The subunit was precipitated by antibody prepared against the aerobic enzyme. Antibody prepared against the subunit also reacted in precipitin tests with the subunit, but not with the native enzyme. The latter antibody nevertheless inhibited the native enzyme but not the nonaerobic isozyme.