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Biomedical subjects

R J Bruce

Publications and source records attributed to R J Bruce.

12 recordsLinked to original sources

Microbial characterization of free floating condensate aboard the Mir space station.

Three samples of humidity condensate that had accumulated behind panels aboard the Russian space station Mir were collected and returned to earth for analysis. As these floating masses of liquid come into contact with the astronauts and the engineering systems, they have the potential to affect both crew health and systems performance. Using a combination of culturing techniques, a wide variety of organisms were isolated included Escherichia coli, Serratia marcescens, and a presumed Legionella species. In addition, microscopic analysis indicated the presence of protozoa, dust mites, and spirochetes. These findings suggest the need for more comprehensive microbial analysis of the environment through the use of new methodologies to allow a more thorough risk assessment of spacecraft.

Animals↗

Microbial colonization of a closed growth chamber during hydroponic cultivation of lettuce.

The goal of this study was to characterize sessile and planktonic microbiota that developed during two successive hydroponic cultures of lettuce in a closed chamber system. Coupons of polyvinyl chloride (PVC) placed in the nutrient solution lines were removed periodically, as were samples of the nutrient solutions and condensate from the air-handling system. The bacteria and fungi present on the coupons and in fluid samples were enumerated by direct plate counts. Disinfecting the hydroponic system with 0.1% hypochlorite and 0.1 N nitric acid reduced the bacterial densities in biofilm samples from 1 x 10(7) CFU/10 cm2 to 1 x 10(1) CFU/10 cm2 and eliminated culturable fungi; Staphylococcus sp., Pseudomonas sp., and Micrococcus sp. survived this procedure. Bacterial and fungal concentrations in all samples returned to predisinfection levels after 2 days of plant growth. Pseudomonas and Acremonium predominated both before and after disinfection. Fungal concentrations never exceeded 7 x 10(2) CFU/10 cm2. The coupon microbiota differed from that of the rhizoplane at harvest. Overall, the greatest numbers of species were found on the rhizoplane samples collected during the second crop. The microbial community changed little during individual crops or between successive crops. Diversity indices remained relatively constant for all samples.

Bacteria↗

Microbiological characterization of a regenerative life support system.

A Variable Pressure Plant Growth Chamber (VPGC), at the Johnson Space Center's (JSC) ground-based Regenerative Life Support Systems (RLSS) test bed, was used to produce crops of soil-grown lettuce. The crops and chamber were analyzed for microbiological diversity during lettuce growth and after harvest. Bacterial counts for the rhizosphere, spent nutrient medium, heat exchanger condensate, and atmosphere were approximately 10(11) Colony Forming Units (CFU) g-1 10(5) CFU ml-1, 10(5) CFU ml-1, and 600 CFU m-3, respectively. Pseudomonas was the predominant bacterial genus. Numbers of fungi were about 10(5) CFU g-1 in the rhizosphere, 4-200 CFU ml-1 in the spent nutrient medium, 110 CFU ml-1 in the heat exchanger condensate, and 3 CFU m-3 in the atmosphere. Fusarium and Trichoderma were the predominant fungal genera.

Bacteria↗

Elicitation of lignin biosynthesis and isoperoxidase activity by pectic fragments in suspension cultures of castor bean.

Suspension cultures of castor bean (Ricinus communis L.) which have been treated with pectic fragment elicitor rapidly accumulate lignin as measured by derivatization with thioglycolic acid. The responsiveness of cultured cells to elicitor is dependent on the stage of culture growth. In 6-day (maximally responsive) cultures, increases in lignin are first evident 3 hours after addition of pectic fragment elicitor with maximal rates of lignin synthesis between 4 and 10 hours. The abundance of lignin in cultures after 12 hours of elicitor treatment is 10- to 20-fold higher than in untreated control cultures and can thereby account for as much as 2% of the dry cell weight. Only intermediate sizes of pectic oligomer are active as elicitors of lignin. Half-maximal accumulation of lignin occurs at 250 to 300 micrograms per milliliter of an optimal elicitor preparation with an average degree of polymerization of seven. We consider the synthesis of lignin in elicited cultures to be a mechanism of plant disease resistance which is induced by the elicitor. Plant peroxidases have been proposed to catalyze the last enzymatic steps in the biosynthesis of both lignin and hydrogen peroxide. Six extracellular isoenzymes of peroxidase (two anionic, designated A1 and A2, and four cationic, designated C2, C3, C4, and C7) are detectable in healthy suspension cultures of castor bean by native gel electrophoresis. Treatment of cultures with elicitor causes substantial changes in the activity of four of these species (A1, C2, C3, and C7). Elicitor treatment also results in the appearance of three new peroxidase isoenzymes that are not readily detectable in healthy cultures (C1, C5, and C6). Increases in the activities of these isoenzymes are concurrent with or slightly precede the accumulation of lignin in elicited 6-day cultures. By 12 hours after addition of elicitor, C1 becomes the most abundant extracellular isoperoxidase. The differential regulation of expression of peroxidase isoenzymes following elicitor treatment suggests that individual isoenzymes of peroxidase may have specific functional roles in the biosynthesis of disease-lignin.

Journal Article↗

Cytokinetics and SCE baselines in rat and human lymphocytes during successive divisions in the presence of different culture media.

Peripheral blood cultures were set up from male rats and humans in TC199, RPMI 1640, and minimal essential medium in the presence of 5-bromo,2-deoxyuridine and harvested at 48, 72, and 96 h. Mitotic indices were compared in the different media at all three harvest times, but cytokinetic patterns and baseline sister chromatid exchange (SCE) frequencies were evaluated exclusively in the 72- and 96-h cultures. In general, lymphocyte division kinetics, as determined by average lymphocyte division (ALD) numbers, were comparable between rat and human lymphocytes cultured in any of the three media and harvested at either of the culture times. The numbers of SCEs were distributed between and within the two systems independent of ALD numbers or the harvest time. Overall, no influence of media was detected on the distribution of SCEs. Despite a number of similarities in growth characteristics between rat and human lymphocytes, the rat lymphocyte test system has distinct advantages over that of the human because of the easy access to rat blood samples and the absence of the many restrictions applicable to humans.

Animals↗

Embolic management of rare hemorrhagic gynecologic and obstetrical conditions.

Severe life-threatening hemorrhage was controlled by angiographic management in 3 patients with bleeding due to unusual gynecological abnormalities and in 1 patient with a rare obstetrically related hemorrhage. Successful management of such rare causes of bleeding emphasizes that early angiographic intervention can, in selected patients, reduce the need for an immediate or subsequent surgical procedure and allow conservative management followed by disease-specific therapy.

Adult↗

Effect of food and water deprivation on the parameters of the mouse bone marrow micronucleus test.

This study reports the influence of acute starvation on spontaneous and cyclophosphamide (CP) induced micronucleus (MN) frequencies in the bone marrow polychromatic erythrocytes (PCE) of CD-1 mice. Groups of mice (5/sex) were deprived of either food alone or food and water for 0, 24, 48 and 72 h prior to sacrifice. Although there was no evidence of a significant increase in MN-PCE frequencies among the starved groups, a significant depressant effect of starvation on erythropoietic activity was observed. Peak levels of CP-induced (40 mg/kg b.w.) MN-PCE's appeared later in male mice deprived of food and water after treatment compared to mice given food and water ad libitum. The results indicate that starvation is detrimental to bone marrow erythropoiesis and that starvation may alter the response of mice to clastogens.

Animals↗

Analyses for in vitro growth conditions, cytokinetics, and sister chromatid exchanges in lymphocytes cultured from the Sprague-Dawley rat.

Peripheral blood samples from Sprague-Dawley rats gave successful lymphocyte growth in GIBCO: 1A, RPMI 1640, and Eagle's minimum essential medium (MEM) culture media. Various growth conditions, cytokinetics, and sister chromatic exchange (SCE) induction were studied using reconstituted GIBCO 1A only. Neither methoxyflurane anesthesia of the rats before sampling nor washing of the cells with phosphate buffered saline affected the mitotic index. Cultures treated with [3H]thymidine showed the lymphocytes entering into DNA synthesis after approximately 24 h. The time at which BUdR (5-bromo-2' deoxyuridine) was added, i.e. 0 vs. 24 h incubation, had minimal effect on the mitotic index of cultures harvested at 48 h. However, when harvest was extended to 72 h, mitotic activity was greater in the cultures treated with BUdR at 24 h. No significant differences in mitotic index and the number of average lymphocyte division were detected in cultures exposed to 0.3 to 0.5 microgram/ml BUdR at 24 h and harvested at 72 h. Although SCE frequencies increased in the presence of BUdR, the baseline level of SCEs was estimated to be 5 to 6/cell. Average generation time of the lymphocytes dividing between 48 and 72 h was 16.5 h. Because of its simplicity of culture and the reproducible nature of its in vitro growth kinetics, the Sprague-Dawley rat lymphocyte is a suitable model for cytogenetic investigations.

Anesthesia, Inhalation↗

The role of feeding rejection in Drosophila mutation assays.

The frequencies of sex-linked recessive lethal mutations recovered in the male post-meiotic germ cells of Drosophila after feeding on solutions containing a mutagen (either 1 mM methyl methanesulfonate, MMS or 2 mM ethyl methanesulfonate, EMS) and 5-bromo-2-deoxyuridine (BUdR) were significantly lower than the frequencies observed after treatment with the mutagen alone. In an attempt to explain the apparent 'protective' effect of BUdR, the feeding behavior of the flies was monitored for differences in the uptake of the mutagen-containing solution in the presence and absence of BUdR. This was accomplished by measuring the uptake of [14C]sucrose. The results indicated that the uptake of the feeding solution is inhibited by the presence of the selected concentration of BUdR (1.0 or 32.5 mM). Such a reduced uptake of the mutagen could alone account for the reduction in mutational yields noticed in treatments containing mutagen + BUdR compared to the ones with the mutagen alone. These results emphasize the need to monitor the feeding behavior of flies in experiments involving adult feeding.

Administration, Oral↗

Elicitation of Casbene Synthetase Activity in Castor Bean : THE ROLE OF PECTIC FRAGMENTS OF THE PLANT CELL WALL IN ELICITATION BY A FUNGAL ENDOPOLYGALACTURONASE.

Endopolygalacturonase isolated from culture filtrates of the fungus Rhizopus stolonifer was shown previously to act as an elicitor of biosynthetic capacity for the antifungal agent, casbene, in castor bean (Ricinus communis L.) seedlings (S.-C. Lee, C.A. West 1981 Plant Physiology 67:633-639). Selective amidation of exposed carboxyl groups of the pure fungal endopolygalacturonase using intermediate activation with a water-soluble carbodiimide under mild conditions leads to inactivation of its enzymic activity. Tests of active and partially inactivated preparations of the enzyme reveal a close correlation between the levels of catalytic and elicitor activities. This suggests that the catalytic activity of the enzyme is necessary for its function as an elicitor. Treatment of the cell-free particulate fraction of homogenates of castor bean seedlings with the active fungal endopolygalacturonase results in the production of a heat-stable, water-soluble component which is highly active as an elicitor of casbene synthetase activity. Several additional lines of evidence, including the susceptibility of the heat-stable elicitor fraction to partial inactivation following prolonged treatment with endopolygalacturonase, indicate that the heat-stable elicitor is most likely a pectic fragment of the plant cell wall and that it is a required intermediate in the process of elicitation of casbene synthetase activity by the fungal endopolygalacturonase.

Journal Article↗