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Biomedical subjects

R C Durley

Publications and source records attributed to R C Durley.

29 records · Page 2Linked to original sources
PubMed1989

Immunoaffinity techniques applied to the purification of gibberellins from plant extracts.

The use of immunoaffinity columns containing anti-gibberellin (GA) antibodies for the selective purification of GAs in plant extracts is described. GA(1), GA(3), GA(4), GA(5), GA(7), and GA(9) conjugates to bovine serum albumin were synthesized and used to elicit anti-GA polyclonal antibodies (Abs) in rabbits. Protein A purified rabbit serum, containing a mixture of anti-GA Abs, was immobilized on matrices of Affi-gel 10 or Fast-Flow Sepharose 4B. Columns of these immunosorbents retained a wide range of C-19 GA methyl esters, but no C-20 GA methyl esters. Quantitative recovery of C-19 GA methyl esters was achieved from the columns, which, after reequilibration in buffer, could be reused up to 500 times. The immunosorbents were tested by examination of extracts from immature soybean and pea seeds. GAs were initially purified by passing the extracts through DEAE-cellulose and concentrating them on octadecylsilica. The extracts were methylated and further purified on the mixed anti-GA immunoaffinity columns. GAs were detected and quantified as methyl esters or methyl ester trimethylsilyl ethers by gas chromatography-mass spectrometry-selected ion monitoring. GA(7) was found in soybean seeds, 17 days after anthesis, at low levels (8.8 nanograms per gram fresh weight). C-19 GAs were examined in cotyledons, embryonic axes, and testae of G2 pea seeds harvested 20 days after anthesis. High levels of GA(20) and GA(29) were found in cotyledons (3580 and 310 nanograms per gram fresh weight, respectively) and embryonic axes (5375 and 1430 nanograms per gram) fresh weight, respectively). Lower levels of GA(9) were found in cotyledons and embryonic axes (147 and 161 nanograms per gram fresh weight, respectively). GA(9) was the major GA of testae at levels of 195 nanograms per gram fresh weight. Trace quantities of GA(20) and GA(51) were also observed in testae.

Journal Article↗
PubMed1989

In vivo metabolism of atrial natriuretic peptide: identification of plasma metabolites and enzymes responsible for their generation.

The in vivo metabolism of atrial natriuretic peptide (ANP) has been studied in the rat after i.v. administration of either [106Phe-14C]- or [126Tyr-125I]-ANP(103-126). Plasma samples containing radioactive peptides were separated by reverse-phase high-performance liquid chromatography. The major plasma metabolites were [125I]Tyr and [14C]Phe for the iodinated and 14C-labeled peptides, respectively. Both peptides had ANP(104/5-126) as a metabolite. Administration of labeled peptide by either bolus or infusion produced the same metabolite profile. To determine which enzymes were responsible for generating these initial metabolites, animals were first dosed with various protease inhibitors before the infusion of [14C]ANP(103-126). The amino-peptidase inhibitor bestatin and the angiotensin converting enzyme inhibitor captopril caused 54 and 66% increases in plasma ANP(103-126), respectively, but no other effects. Administration of the endopeptidase 24.11 inhibitor thiorphan led to a 158% increase of ANP(103-126) in plasma and an 11-fold increase in ANP(104/5-126). The latter metabolite could be selectively decreased by pretreatment with bestatin in combination with thiorphan. The results demonstrate that the initial plasma metabolites of ANP(103-126) are due to the activity of endopeptidase 24.11, a bestatin-sensitive aminopeptidase, and a carboxypeptidase. The plasma clearance of the peptide is probably also due to cellular binding and uptake in combination with glomerular filtration as very few plasma metabolites were observed even at very high rates of ANP(103-126) infusion.

Animals↗
PubMed1987

The Physiological Basis for Cytokinin Induced Increases in Pod Set in IX93-100 Soybeans.

Previous investigations have shown the feasibility of increasing pod number on legumes by the application of 6-benzylaminopurine (BA) directly to the raceme. These investigations were designed to determine what reproductive parameter was affected by cytokinin application, and if these applications were overcoming a deficiency in root-produced cytokinins during late flowering. Five individual main stem racemes on greenhouse grown soybeans (Glycine max L. Merr.) were treated with 2 millimolar BA. A single application of BA when pods appeared at 25 to 50% of the proximal floral positions resulted in a 58% increase in pod set due primarily to a 33% reduction in floral abscission. Applications of BA at later intervals also resulted in significant reductions in total abscission. When three applications of BA were imposed on the upper five nodes of field grown soybeans, total pod number and seed weight were significantly increased in this section of the canopy by 27 and 18%, respectively. Throughout the flowering period, root pressure exudate was sampled for the subsequent separation and quantification of zeatin, dihydrozeatin, zeatin riboside, dihydrozeatin riboside, and isopentenyladenine. Total cytokinin flux peaked from 0 to 9 days after flowering began, and then dropped to one-half of this level by 15 days postanthesis. The probability that a flower would initiate a pod was directly related to the concentration of total cytokinins present in the exudate when the flower opened.

Journal Article↗
PubMed1979

Metabolism of Tritiated Gibberellin A(20) in Immature Seeds of Dwarf Pea, cv. Meteor.

Tritium-labeled gibberellin A(20) ([(3)H]GA(20)) was applied via the pedicel to immature pods and seeds of dwarf peas and three harvests were made at days 5, 10, and 23 (mature) after application. Of the five metabolites of [(3)H]GA(20), the three in highest yield were GA(29), an alpha,beta-unsaturated ketone, and a compound (B), whose structure was only tentatively assigned. The metabolic sequence GA(20) --> GA(29) --> compound B --> the ketone was indicated. The amount of [(3)H]GA(29) in both seeds and pods was highest at day 5 and declined to its lowest level at maturity. The amount of the [(3)H]ketone in the seed increased with time to its highest level at maturity. It is suggested that compound B and the ketone represent the major pathway of catabolism of GA(29), a 2beta-hydroxylated GA of low biological activity, and that the ketone is not metabolized, or only slowly metabolized, during seed maturation.

Journal Article↗
PubMed1977

Biosynthesis of gibberellins in Gibberella fujikuroi. Gibberellin A47.

Two minor radioactive products in cultures of G. fujikuroi strain ACC917 supplemented with labeled gibberellin A4 have been identified as the 2alpha-hydroxy derivative, gibberellin A47, and the hydrate, gibberellin A2. In addition, evidence was obtained by combined gas chromatography--mass spectrometry for the formation of 3-O-acetylgibberellin A1,3-O-acetylgibberellin A3, and gibberellin A20 by the culture. The time course of gibberellin synthesis in defined and complex media, as well as changes in the relative amounts of radioactive gibberellins formed when [4C]gibberellin A4 was added to cultures at different ages, suggest that the composition of the gibberellin mixture produced is determined by the balance between synthesis and decay of key enzymes in the branching pathway.

Gibberella↗
PubMed1976

Effects of Light, Abscisic Acid, and N-Benzyladenine on the Metabolism of [H]Gibberellin A(4) in Seeds and Seedlings of Lettuce, cv. Grand Rapids.

Gibberellin A(4) (GA(4)) can substitute for light in the germination of Grand Rapids lettuce seeds. Seeds imbibed in [(3)H]GA(4) do not convert this to other GAs prior to, or immediately following, visible germination: thus GA(4) alone can promote radicle expansion. Abscisic acid inhibited [(3)H]GA(4)-induced germination, but did not significantly affect [(3)H]GA(4) uptake or metabolism during germination. (6)N-benzyladenine overcame the inhibitory effect of abscisic acid and increased [(3)H]GA(4) uptake, although radicle emergence was delayed somewhat.During hypocotyl extension there was a large conversion of [(3)H]GA(4) to [(3)H]GA(1) in light or darkness, the major conversion site being the growing root. Hypocotyls of dark-grown seedlings contained more [(3)H]GA(1) than those of light-grown seedlings. The apparent inability of exogenous GA(1) to promote greater hypocotyl extension than GA(4) is related to its poorer uptake. Abacisic acid markedly inhibited hypocotyl expansion, root growth, and the conversion of [(3)H]GA(4) to [(3)H]GA(1).

Journal Article↗
PubMed1976

Endogenous Gibberellins of Pine Pollen: III. Conversion of 1,2-[H]GA(4) to Gibberellins A(1) and A(34) in Germinating Pollen of Pinus attenuata Lemm.

Gibberellins A(1) and A(34) (possibly A(2)) were found as products of metabolism of 1,2-[(3)H]GA(4) during germination of Pinus attenuata pollen. The conversion from GA(4) to GA(1) and GA(34) occurred as hydroxylations at atoms C-13 and C-2 of the ent-gibberellane skeleton, respectively. Percentage interconversion of the GA(4) absorbed was in the range of 0.15 to 0.43% for GA(1) and 1.54 to 3.22% for GA(34). Identifications were made on a gas-liquid chromatograph with radioactive monitoring by comparison with standards.

Journal Article↗
PubMed1976

Metabolism of [H]Gibberellin A(20) in Light- and Dark-grown Tobacco Callus Cultures.

The growth of tobacco callus in culture (previously shown to contain gibberellin [GA]-like substances), and its ability to metabolize [(3)H]-GA(20) were examined. Growth rates, in the presence and absence of exogenously applied GA, were examined in light and dark conditions. Dark-grown callus grew at a much faster rate than light-grown and [(3)H]GA(20) was metabolized much more rapidly in darkness than in light.[(3)H]GA(1) was identified by combined gas-liquid chromatography/mass spectrometry as a major product of [(3)H]GA(20), and was found to be a more potent promoter of tobacco callus growth than GA(20).

Journal Article↗
PubMed1974

Metabolism of Tritiated Gibberellin A(9) by Shoots of Dark-grown Dwarf Pea, cv. Meteor.

Tritium-labeled gibberellin A(9) ((3)H-GA(9)) was metabolized by etiolated shoots of dwarf pea (Pisum sativum cv. Meteor) to GA(20), GA(10), 2,3-dihydro-GA(31), and a number of highly polar, acidic GA-like substances. Identifications were made by gasliquid radiochromatography and combined gas chromatography-mass spectrometry. Kinetic studies showed that GA(30) and 2,3-dihydro-GA(31) were produced within 5 hours following (3)H-GA(9) application to pea shoots. The polar GA-like substances were produced between 5 and 10 hours after (3)H-GA(9) application. Levels of GA(10) increased with time, and since no GA(10) was produced during the purification procedures, GA(10) was, in all probability, produced from (3)H-GA(9) within the plant tissue. The radioactive interconversion products produced by pea from (3)H-GA(9) have chromatographic properties similar to biologically active GA-like substances present in etiolated shoots of dwarf pea. Large scale applications of (3)H-GA(9) with very low specific activity to etiolated pea shoots showed that the radioactivity of the interconversion products was correlated exactly with biological activity as assayed by dwarf rice (Oryza sativa cv. Tan-ginbozu).

Journal Article↗