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D P Roberts

Publications and source records attributed to D P Roberts.

At least 19 recordsLinked to original sources
PubMed2001

Analysis of fungal communities by sole carbon source utilization profiles.

A simple method for characterization of fungal communities in environmental samples was developed. Dilute suspensions of samples in 0.2% agar containing three different antibiotics were pipetted into 96-well plates (Biolog SF-N) containing a diverse collection of 95 different carbon sources. The plates were incubated for 4-12 days at 22 degrees C and the absorbance measured at 650 nm. Canonical variates analysis was then used to analyze the multivariate data. This method allowed fungal communities in rhizosphere soil of corn and soybean to be distinguished according to soil and plant type. Data taken at a single time-point, which varied greatly in total absorbance of the plate, separated rhizosphere samples primarily by soil type. When multiple time-points were combined to keep the total absorbance constant, differences in substrate utilization patterns due to different plant types could be distinguished. The method was also applicable to analysis of phylloplane and compost fungal communities. This method is readily applied to large numbers of samples and should be useful for community analysis in a variety of agricultural and ecological studies.

Carbon↗
PubMed2001

Antihypertensive therapy in critical occupations: studies with an angiotensin II antagonist.

OBJECTIVE: This study was performed to establish whether an angiotensin II antagonist modulates daytime sleepiness or performance. METHODS: The central effects of losartan (50 and 100 mg), as well as amlodipine (5 and 10 mg) and promethazine (10 mg), were studied in six healthy volunteers (aged between 20 and 27 (mean 24) yr). Digit symbol substitution, tracking, vigilance, immediate and short-term memory recall, complex reaction time, together with objective (daytime sleep latencies) and subjective measures of sleepiness, were measured 1.0 h before and 0.5, 2.0, 3.5, 5.0, 6.5, and 8.0 h after ingestion. The study was placebo-controlled and double-blind with a six-way crossover design. RESULTS: No changes in performance or in measurements related to sleepiness were observed with losartan (50 and 100 mg). Amlodipine (10 mg) impaired immediate memory and increased subjective sleepiness at 8.0 h (p < 0.05). Promethazine (10 mg) impaired vigilance and immediate memory recall and increased reaction time (basic and complex) from 2.0 to 6.5 h after ingestion, impaired tracking from 3.5 to 6.5 h and increased subjective sleepiness from 2.0 to 8.0 h (p < 0.05). CONCLUSION: The study suggests that an angiotensin II antagonist could prove useful for the treatment of hypertension in aircrew.

Adult↗
PubMed2000

Importance of pfkA for rapid growth of Enterobacter cloacae during colonization of crop seeds.

Enterobacter cloacae A-11 is a prototrophic, glycolytic mutant of strain 501R3 with a single transposon insertion in pfkA. The populations of strain A-11 on cucumber and radish seeds were smaller than the populations of strain 501R3 in natural soil, but the populations of these two strains on pea, soybean, sunflower, and sweet corn seeds were similar (D. P. Roberts, P. D. Dery, I. Yucel, J. Buyer, M. A. Holtman, and D. Y. Kobayashi, Appl. Environ. Microbiol. 65:2513-2519, 1999). The net effect of the mutation in pfkA in vitro was a shift from rapid growth on certain carbohydrates detected in seed exudates to much slower growth on other carbohydrates, amino acids, and organic acids. The impact of the mutation in pfkA was greatest on the growth rate of E. cloacae on the seeds that released the smallest quantities of fructose, other carbohydrates, and amino acids. Corn, pea, soybean, and sunflower seeds released total amounts of carbohydrates and amino acids at rates that were approximately 10- to 100-fold greater than the rates observed with cucumber and radish seeds for the first 24 h after inhibition began. The growth rate of strain A-11 was significantly less (50% less) than the growth rate of strain 501R3 on radish seeds, and the growth rate of strain A-11 was too low to estimate on cucumber seeds in sterile sand for the first 24 h after inhibition began. The growth rate of strain A-11 was also significantly lower on soybean seeds, but it was only 17% lower than the growth rate of strain 501R3. The growth rates of strains 501R3 and A-11 were similar on pea, sunflower, and corn seeds in sterile sand for the first 30 h after imbibition began. Large reductions in the growth rates of strain A-11 on seeds were correlated with subsequent decreased levels of colonization of seeds compared to the levels of colonization of strain 501R3. The strain A-11 populations were significantly smaller than the strain 501R3 populations only on radish and cucumber seeds. The mutation in pfkA appears to decrease the level of colonization by E. cloacae for seeds that release small quantities of reduced carbon compounds by decreasing the size of the pool of compounds that support rapid growth by this bacterium.

Amino Acids↗
PubMed1999

Role of pfkA and general carbohydrate catabolism in seed colonization by Enterobacter cloacae.

Enterobacter cloacae A-11 is a transposon mutant of strain 501R3 that was deficient in cucumber spermosphere colonization and in the utilization of certain carbohydrates (D. P. Roberts, C. J. Sheets, and J. S. Hartung, Can. J. Microbiol. 38:1128-1134, 1992). In vitro growth of strain A-11 was reduced or deficient on most carbohydrates that supported growth of strain 501R3 but was unaffected on fructose, glycerol, and all amino acids and organic acids tested. Colonization by strain A-11 was significantly reduced (P </= 0.05) for cucumber and radish seeds compared to that of strain 501R3, but colonization of pea, soybean, sunflower, and sweet corn seeds was not reduced. Pea seeds released several orders of magnitude more total carbohydrates and amino acids than cucumber and radish seeds and approximately 4,000-fold more fructose. Fructose was the only carbohydrate detected in the seed exudates which supported wild-type levels of in vitro growth of strain A-11. Soybean, sunflower, and sweet corn seeds also released significantly greater amounts of fructose and total carbohydrates and amino acids than cucumber or radish seeds. The exogenous addition of fructose to cucumber and radish seeds at quantities similar to the total quantity of carbohydrates released from pea seeds over 96 h increased the populations of strain A-11 to levels comparable to those of strain 501R3 in sterile sand. Molecular characterization of strain A-11 indicated that the mini-Tn5 kanamycin transposon was inserted in a region of the genome with significant homology to pfkA, which encodes phosphofructo kinase. A comparison of strain A-11 with Escherichia coli DF456, a known pfkA mutant, indicated that the nutritional loss phenotypes were identical. Furthermore, the pfkA homolog cloned from E. cloacae 501R3 complemented the nutritional loss phenotypes of both E. coli DF456 and E. cloacae A-11 and restored colonization by strain A-11 to near wild-type levels. These genetic and biochemical restoration experiments provide strong evidence that the quantities of reduced carbon sources found in seed exudates and the ability of microbes to use these compounds play important roles in the colonization of the spermosphere.

Carbohydrate Metabolism↗
PubMed1996

In vitro analysis of the role of glucose oxidase from Talaromyces flavus in biocontrol of the plant pathogen Verticillium dahliae.

Culture filtrates from Talaromyces flavus grown on glucose contained high levels of glucose oxidase activity, while culture filtrates from T. flavus grown on xylan contained negligible glucose oxidase activity. Culture filtrates from T-flavus grown on both media contained complex protein profiles. However, only culture filtrates from T. flavus grown on glucose inhibited germination of microsclerotia of Verticillium dahliae in in vitro inhibition assays. A polyclonal antiserum preparation, pABGO-1, raised against purified glucose oxidase from T. flavus was highly specific for glucose oxidase. Only one protein band in culture filtrates (from glucose medium), migrating at 71 kDa, was detected in Western blots (immunoblots) with this antiserum. This band comigrated with purified glucose oxidase. No bands were detected in culture filtrates from the xylan medium. Glucose oxidase was removed via immunoprecipitation from culture filtrates of T. flavus grown in glucose medium, resulting in filtrates which no longer inhibited in vitro microsclerotial germination. When glucose oxidase-depleted filtrates were amended with purified glucose oxidase from T. flavus, the ability to kill microsclerotia in vitro was restored to original levels. We conclude that glucose oxidase is the only protein in culture filtrates of T. flavus responsible for inhibition of germination of microsclerotia of V. dahliae.

Animals↗
PubMed1996

Isolation and modulation of growth of a colonization-impaired strain of Enterobacter cloacae in cucumber spermosphere.

Enterobacter cloacae A-46 was isolated for use in an environmental containment strategy for genetically modified derivative strains with enhanced biocontrol activity. The population of E. cloacae A-46, a transposon mutant of the plant-beneficial bacterium E. cloacae 501R3, increased 10-fold (significant increase at P < or = 0.05) in cucumber spermosphere when applied to cucumber seeds along with casamino acids. In contrast, strain A-46 was incapable of proliferation in cucumber spermosphere in the absence of casamino acids. Populations of strain A-46 also failed to increase in corn, cowpea, sunflower, and wheat spermospheres in the absence of casamino acids, while populations of strain 501R3 increased 3162-, 512-, 1698-, and 93-fold, respectively. In addition, the persistence of strain A-46 in corn, cucumber, and sunflower rhizospheres and in natural soil was greatly reduced compared with the parental strain 501R3.

Cucumis sativus↗
PubMed1988

Development of a passive device for freezing large amounts of transplantable skin at one time in a -70 degrees C mechanical refrigerator.

A simple device has been developed for the simultaneous cooling of up to 9120 cm2 of allograft skin in a flat package format. The device, named an insulated alternating-offset heat sink device, is composed of a stack of interleaved layers of 2.0-mm-thick packets of skin and 3.18-mm-thick aluminum heat sinks (each 33.0 cm long by 22.9 cm wide). Four skin packets are placed in a single layer on each heat sink plate, and the number of plates can be varied to accommodate different numbers of skin packets. Every heat sink protrudes 6.3 cm of its 33.0-cm length beyond the skin packets to make a fin for heat convection, but adjacent plates alternate the direction of their fin protrusion so that the layers of plates alternate in their 6.3-cm offset. Insulation layers of 2.54-cm-thick expanded polystyrene are placed on the exposed surfaces of the top and bottom heat sinks in the stack, and the stack is held together by rubber bands. The device is cooled in a -70 degrees C mechanical refrigerator. Maximal cooling rates of -1.8 degrees C min-1 are obtained for both 6- and 11-plate devices, and -3.0 degrees C min-1 for a 2-plate device. The exothermic temperature plateaus associated with skin cooled in these devices are 1.5-1.8 min in duration. Skin cooled by this technique maintains levels of glucose oxidation similar to those associated with skin cooled by liquid nitrogen vapor at a controlled rate of -1 degree C min-1, provided rapid warming is employed after -70 degrees C storage. The development of this device provides a method for the simple, low-cost cryopreservation of the large amounts of allograft skin obtained from a cadaveric donor.

Cell Survival↗
PubMed1988

Analysis of the Pseudomonas solanacearum polygalacturonase encoded by pglA and its involvement in phytopathogenicity.

A major endopolygalacturonase excreted by Pseudomonas solanacearum was purified to greater than 95% homogeneity and shown to have an isoelectric point of 9.0 and a subunit molecular mass of 52 kilodaltons (kDa). The gene encoding this enzyme (pglA) was isolated from a genomic library of P. solanacearum DNA based on its expression in Escherichia coli and shown to be contained on a 1.8-kilobase DNA fragment. The identity of the pglA gene product and the 52-kDa polygalacturonase was demonstrated by immunoadsorption and isoelectric focusing experiments. The cloned pglA gene was apparently expressed from its own promoter in E. coli and its product was partially secreted into the periplasm. The pglA gene was insertionally inactivated in vitro and used to mutate the chromosomal pglA gene of P. solanacearum by marker exchange mutagenesis. The resulting mutant strain was deficient in production of the 52-kDa polygalacturonase and took twice as long to wilt and kill tomato plants as the wild-type parent in plant bioassay experiments. Complementation in trans with the wild-type cloned pglA gene restored virulence to near wild-type levels. The data indicate that the pglA gene is important, but not absolutely necessary, for pathogenesis.

Escherichia coli↗
PubMed1988

Cloning of the egl gene of Pseudomonas solanacearum and analysis of its role in phytopathogenicity.

The egl gene of Pseudomonas solanacearum was cloned on a cosmid and expressed in Escherichia coli. Restriction endonuclease mapping, transposon mutagenesis, and subclone analysis showed that the egl gene was located on a 2.7-kilobase XhoI-SalI P. solanacearum DNA fragment. Immunoabsorption experiments and sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis showed that the egl gene encodes the 43-kilodalton endoglucanase that is the major excreted endoglucanase of P. solanacearum. In E. coli, the egl gene appeared to be expressed from its own promoter, but its product was restricted to the cytoplasm. The cloned egl gene was mutagenized with Tn5 and used to specifically mutate the chromosomal egl gene of P. solanacearum by site-directed mutagenesis. The resultant mutant was identical to the wild-type strain in production of extracellular polysaccharide and extracellular polygalacturonase as well as several other excreted proteins but produced at least 200-fold less endoglucanase. This mutant strain was significantly less virulent on tomato than the wild-type strain in plant bioassay experiments. Virulence of the endoglucanase-deficient strain was restored to near wild-type levels by complementation in trans with the cloned egl gene, indicating that the egl gene is important but not absolutely required for pathogenesis.

Cellulase↗
PubMed1987

Adapting personal computers for use by high-level quadriplegics.

A communication instrument has been developed for use by a quadriplegic person who is limited solely to eye motion as a control input. The instrument uses an infrared-emitting diode/phototransistor sensor mounted on the frame of a pair of eyeglasses to transform deliberate lateral eye motion into a contact-closure output form. A software program has also been developed for use with this instrument to display letters of the alphabet in a series of six lines. The lines are slowly scrolled until the desired line is reached, then stopped by a lateral eye motion. The scrolling then continues for each character in that line until the desired character is reached. Again, lateral eye movement causes the scrolling to stop and the character becomes part of a string. This process continues until a command character is selected whereby the string can be sent to a printer, spoken through a speech synthesizer, or erased.

Adult↗
PubMed1986

Requirement for two or more Erwinia carotovora subsp. carotovora pectolytic gene products for maceration of potato tuber tissue by Escherichia coli.

Several genes encoding enzymes capable of degrading plant cell wall components have been cloned from Erwinia carotovora subsp. carotovora EC14. Plasmids containing cloned EC14 DNA mediate the production of endo-pectate lyases, exo-pectate lyase, endo-polygalacturonase, and cellulase(s). Escherichia coli strains containing one of these plasmids or combinations of two plasmids were tested for their ability to macerate potato tuber slices. Only one E. coli strain, containing two plasmids that encode endo-pectate lyases, exo-pectate lyase, and endo-polygalacturonase, caused limited maceration. The pectolytic proteins associated with one of these plasmids, pDR1, have been described previously (D. P. Roberts, P. M. Berman, C. Allen, V. K. Stromberg, G. H. Lacy, and M. S. Mount, Can. J. Plant Pathol. 8:17-27, 1986) and include two secreted endo-pectate lyases. The second plasmid, pDR30, contains a 2.1-kilobase EC14 DNA insert that mediates the production of an exo-pectate lyase and an endo-polygalacturonase. These enzymes are similar in physicochemical properties to those produced by EC14. Our results suggest that the concerted activities of endo-pectate lyases with endo-polygalacturonase or exo-pectate lyase or both cause maceration.

Cell Wall↗
PubMed1980

Holography.

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Forecasting↗
PubMed1976

Oral carbenicillin treatment of uncomplicated gonorrhea in men and women.

In a prospective controlled study of 290 men and women with uncomplicated gonorrhea, carbenicillin was shown to be less than 90% effective. Positive cultures were the basis for initial diagnosis and therapeutic failure in both sexes. Carbenicillin was evaluated in three different schedules: 5gm p.o., 10 gm p.o, and 2gm IM Plus 5GM p.o. The three schedules were equally effective. Eleven of 20 failures in women occurred in infection of the rectum. MICs of carbenicillin ranged from 0.025% to 1.0 mug/ml with a mean of 0.29 mug/ml.

Administration, Oral↗
PubMed1972

Treatment of gonorrhea with spectinomycin hydrochloride: comparison with standard penicillin schedules.

Spectinomycin hydrochloride, a new parenteral antibiotic prepared from Streptomyces spectabilis, was compared with standard U.S. Public Health Service-recommended dosages of aqueous procaine penicillin G in the treatment of uncomplicated gonorrhea in 353 men and 314 women. Of the 314 women, 130 had a pretreatment positive rectal culture. All diagnoses were proven by culture on Thayer-Martin selective medium. Minimal inhibitory concentrations of both drugs were determined. Single doses of 2 and 4 g of spectinomycin were compared with 2.4 million units of procaine penicillin in males and with both 2.4 and 4.8 million units of procaine penicillin in females. Both spectinomycin schedules, 2.4 million units of penicillin in males and 4.8 million units of penicillin in females, resulted in cure rates in excess of 90%. There were no failures at the rectal site only in those women with positive rectal cultures. There was no advantage to using the larger amount of spectinomycin in either sex.

Antibiotics, Antineoplastic↗