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C Bryant

Publications and source records attributed to C Bryant.

At least 55 records · Page 3Linked to original sources
PubMed1991

Purification and characterization of an oxygen-insensitive NAD(P)H nitroreductase from Enterobacter cloacae.

The reductive products of several nitroaromatic compounds have been found to be toxic, mutagenic, and carcinogenic. The nitroreductases present in intestinal microflora have been implicated in the biotransformation of these compounds to their deleterious metabolites. A "classical" nitroreductase has been purified from Enterobacter cloacae 587-fold using a protocol which yields approximately 1 mg of purified nitroreductase from 10 liters of cell culture. An analysis of the physical properties of the nitroreductase indicates that the enzyme is active as a monomer with a calculated molecular mass of 27 kDa. FMN has been identified as a required flavin cofactor and is present at a stoichiometry of 0.88 mol of FMN bound/mol of active enzyme. The enzyme was found capable of reducing nitrofurazone under aerobic conditions indicating that the mechanism involves an obligatory two-electron transfer. Thus, this enzyme can be classified as an oxygen-insensitive nitroreductase. The purified nitroreductase can utilize either NADH or NADPH as a source of reducing equivalents and can reduce a variety of nitroaromatic compounds including nitrofurans and nitrobenzenes as well as quinones. Studies in which the rates of nitroreduction for a series of para substituted nitrobenzene derivatives were determined suggest that a linear free energy relationship exists between the rate and the redox midpoint potential of the substrate.

Bacterial Proteins↗
PubMed1991

Cloning, nucleotide sequence, and expression of the nitroreductase gene from Enterobacter cloacae.

The "classical" nitroreductases of enteric bacteria are flavoproteins which catalyze the reduction of a variety of nitroaromatic compounds to metabolites which are highly toxic, mutagenic, or carcinogenic. The gene for the nitroreductase Enterobacter cloacae has now been cloned using an antibody specific to this protein. The nucleotide sequence of the structural gene and flanking regions are reported. Sequence analysis indicates that this gene belongs to a gene family of flavoproteins which have not been previously described. Analysis of the 5'-untranslated region reveals the presence of putative regulatory elements which may be involved in the modulation of the expression of this enzyme. The cloned gene was placed under the control of a T7 promoter for overexpression of the protein in Escherichia coli. The expressed recombinant protein was purified to homogeneity and exhibited physical, spectral, and catalytic properties identical to the protein isolated from E. cloacae.

Amino Acid Sequence↗
PubMed1991

Cellular localization of ovarian prostaglandin endoperoxide synthase during pseudopregnancy in the rat.

The specific cellular localization of prostaglandin endoperoxide (PGH) synthase, the enzyme responsible for initiating the conversion of arachidonic acid to prostaglandins, was studied throughout pseudopregnancy in the rat. Pseudopregnancy was induced by administration of eCG (20 IU) to immature, 27-day-old rats followed by hCG injection (10 IU) on Day 29. Animals were necropsied on Days 1 (Day 1 = 1 day post hCG), 5, 9, and 13 of pseudopregnancy. Ovaries were removed and processed for cellular identification of PGH synthase by immunohistochemistry. Immunoreactive PGH synthase was distributed throughout the CL at each of the 4 different days of pseudopregnancy, with the majority of the luteal cells exhibiting varying degrees of staining. The connective tissue centrum of the CL, however, lacked PGH synthase immunoreactivity. Quantitative assessment of the immunostaining distribution was accomplished with a computer-based image analysis program (Kontron IBAS). Results are expressed as the percentage of a digitized luteal area that contained intense immunoreactivity between Day 1 (0.6 +/- 0.2% immunoreactive area) and Day 5 (16.8 +/- 2.7%) of pseudopregnancy. The area of luteal immunostaining was similar on Day 5 and Day 9 (16.8 +/- 2.7% and 14.7 +/- 2.0%, respectively) followed by a decrease (p less than 0.05) in immunoreactivity on Day 13 (9.1 +/- 2.2%). The region of the CL adjacent to the germinal epithelium had an increase (p less than 0.01) in PG synthase staining distribution compared to the region of the CL adjacent to the ovarian medulla on all days of pseudopregnancy except Day 1. These findings demonstrate that PGH synthase is present in the rat CL throughout pseudopregnancy.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1991

Free radical generation and the course of primary infection with Nippostrongylus brasiliensis in congenitally athymic (nude) rats.

The course of primary infections with Nippostrongylus brasiliensis was followed in nude (CBH/R nu/nu) and heterozygote (CBH/R nu/+) rats. In both nude and heterozygote rats peak egg production by N. brasiliensis occurred on days 7 and 8 post-infection. However, whereas in heterozygote rats egg production declined rapidly thereafter and ceased completely by day 14 post-infection, in nude rats high numbers of N. brasiliensis eggs were still seen on day 27 post-infection, when the experiment was terminated. In comparison with the nude rats, heterozygote rats had a 5-fold greater loss of weight by day 9 post-infection and a 4-fold higher incidence of diarrhoea. Furthermore, infected heterozygote rats became anaemic whereas uninfected rats and infected nude rats showed no evidence of anaemia. Free radical generation was measured in infected (9 days) and uninfected rats. Leucocytes from infected heterozygote rats were able to generate copious quantities of free radicals in response to N. brasiliensis whereas leucocytes from infected nude rats produced only slightly more free radicals than uninfected rats. Thus, worm rejection, weight loss, diarrhoea, anaemia and free radical generation in response to N. brasiliensis infection are all T-cell dependent events.

Analysis of Variance↗
PubMed1990

The role of the host in the regulation of end-product formation in two strains of the rat tapeworm, Hymenolepis diminuta.

Individual worms from rats infected with different strains of Hymenolepis diminuta were incubated in vitro and the products lactate, succinate, acetate and ammonia assayed. Variability in excretion was not confined to differences between strains. Two metabolic types were identified. Where succinate was above 20 mumol g-1 h-1, lactate excretion was low. Where succinate was not detected, lactate excretion was high. Acetate excretion was variable. Lactate and ammonia excretion were positively correlated. All worms from one rat were of the same type but could be of either type from different rats. The host strain had no effect. A relationship was shown between lactate excretion and the number of worms from a standard inoculum present at 21 days of infection. The incidence of high lactate excretion was increased in worms from secondary infections. Components of the host immune response may thus exert effects on the metabolism of H. diminuta, manifest as shifts in emphasis on cytosolic and mitochondrial metabolism.

Acetates↗
PubMed1990

The third world goes to town: will U.S. policy catch up?

"Third World cities are growing so fast that in the 21st century over half the world's population will be urban. This shift will need to be recognized by all aid donors, and particularly by the USA. Sustainable development will be possible only if Third World governments--central and local--can be assisted to meet the challenge of this massive rural to urban transition."

Americas↗
PubMed1990

Ovarian prostaglandin endoperoxide synthase: cellular localization during the rat estrous cycle.

The specific cellular localization of prostaglandin endoperoxide (PGH) synthase was studied throughout the rat estrous cycle. Animals were necropsied at 1300 h on each day of the 4-day cycle, and an additional group was necropsied at 2300 h on proestrus. Ovaries were removed and processed for cellular identification of PGH synthase by immunohistochemistry. At all stages of the cycle, intense immunostaining was observed in newly formed corpora lutea. Luteal cells were immunoreactive, but the connective tissue centrum was unstained. Interstitial tissue contained heavily labeled cells, whereas the germinal epithelium exhibited faint staining. During estrus, metestrus, and diestrus, thecal cells from preantral and antral follicles contained PGH synthase immunoreactivity, but granulosa cells were unstained. Faint staining of mural granulosa cells was observed first in 78% of preovulatory follicles (less than 400-microns diameter) in ovaries collected on the afternoon of proestrus. After the luteinizing hormone surge, 95% of the preovulatory follicles exhibited PGH synthase staining. The percentage of immunoreactive granulosa cells in these preovulatory follicles increased 4-fold in ovaries collected at 2300 h on proestrus. The presence of ovarian PGH synthase throughout the rat estrous cycle and the changes in cellular localization may reflect the potential role of PGs in follicular and luteal function.

Animals↗
PubMed1989

Genetic characterization of three strains of Hymenolepis diminuta at 39 enzyme loci.

The technique of allozyme electrophoresis was applied to three strains of Hymenolepis diminuta to distinguish between three hypotheses [(1) multiple species, (2) genetically distinct founder stocks and (3) response to differential selection among similar stocks] proposed to account for metabolic differences among strains. There was no evidence from the 39 enzyme loci established that the three strains represented more than one species. In the absence of knowledge of the population structure of H. diminuta in the wild, electrophoretic data herein could not distinguish between the latter hypotheses. Nevertheless, all three strains were distinguishable on electrophoretic profiles and allelic similarities between strains question the view of their proposed origins.

Animals↗
PubMed1989

Free radical generation during primary infections with Nippostrongylus brasiliensis.

The production of free radicals during infection of the rat with Nippostrongylus brasiliensis was investigated. Lipid peroxidation, which is the best documented effect of free radicals, was monitored in the small intestines of infected rats by measurement of malonyldialdehyde and was found to be increased at the time of worm rejection. The capacity of peritoneal leucocytes to produce free radicals, as measured by chemiluminescence, was monitored in rats infected with different doses of N. brasiliensis. Rejection of N. brasiliensis from rats infected with 6000 third-stage larvae (L3) began 2 days earlier than in rats infected with only 600 L3. Maximal free radical generation also occurred 2 days earlier and was quantitatively greater in rats infected with 6000 L3. Free radical generation by leucocytes in response to live adult N. brasiliensis was enhanced by plasma from infected rats indicating the existence of a plasma-borne factor responsible for the initiation of free radical generation in response to N. brasiliensis.

Animals↗
PubMed1989

The effect of antioxidants on the rejection of Nippostrongylus brasiliensis.

The effect of the antioxidants, butylated hydroxy anisole (BHA) and vitamin E, on the rejection of Nippostrongylus brasiliensis from the small intestine of the rat was investigated. Worm expulsion was inhibited by BHA. Malonyldialdehyde production in the small intestines and free radical generation by peritoneal leucocytes from infected rats were also inhibited by BHA. Vitamin E, although inhibiting malonyldialdehyde production, did not prevent worm expulsion. Significantly, vitamin E was much less effective than BHA at reducing free radical generation by rat leucocytes in response to N. brasiliensis.

Animals↗
PubMed1986

Effects of guanidine hydrochloride on the refolding kinetics of denatured thioredoxin.

The effect of guanidine hydrochloride concentration on the kinetics of the conformational change of Escherichia coli thioredoxin was examined by using fluorescence, absorbance, circular dichroic, and viscosity measurements. Native thioredoxin unfolds in a single kinetic phase whose time constant decreases markedly with increasing denaturant concentration in the denaturation base-line zone. This dependency merges with the time constant of the slowest refolding kinetic phase at the midpoint of the equilibrium transition in 2.5 M denaturant. The time constant of the slowest refolding phase becomes denaturant independent below 1 M denaturant in the native base-line region. The denaturant-independent slowest refolding phase has an activation energy of 16 kcal/mol and is generated in the denatured base-line zone in a denaturant-independent reaction having a time constant of 19 s at 25 degrees C. The fractional amplitude of the slowest refolding phase diminishes in the native base-line zone to a minimum value of 0.25. This decrease is accompanied by an increase in the fractional amplitudes of two faster refolding kinetic phases, an increase describing a sigmoidal transition centered at about 1.6 M denaturant. Manual multimixing measurements indicate that only the slowest refolding kinetic phase generates a product having the stability of the native protein. We suggest that the two faster refolding phases reflect the transient accumulation of folding intermediates which can contain a nonnative isomer of proline peptide 76.

Bacterial Proteins↗