Biomedical subjects

A M Reeve

Publications and source records attributed to A M Reeve.

17 recordsLinked to original sources

PubMed1998

Purification, characterization, and cloning of an S-adenosylmethionine-dependent 3-amino-3-carboxypropyltransferase in nocardicin biosynthesis.

S-Adenosylmethionine:nocardicin 3-amino-3-carboxypropyltransferase catalyzes the biosynthetically rare transfer of the 3-amino-3-carboxypropyl moiety from S-adenosylmethionine to a phenolic site in the beta-lactam substrates nocardicin E, F, and G, a late step of the biosynthesis of the monocyclic beta-lactam antibiotic nocardicin A. Whereas a number of conventional methods were ineffective in purifying the transferase, it was successfully obtained by two complementary affinity chromatography steps that took advantage of the two substrate-two product reaction scheme. S-Adenosylhomocysteine-agarose selected enzymes that utilize S-adenosylmethionine, and a second column, nocardicin A-agarose, specifically bound the desired transferase to yield the enzyme as a single band of 38 kDa on a silver-stained SDS-polyacrylamide gel. The transferase is active as a monomer and exhibits sequential kinetics. Further kinetic characterization of this protein is described and its role in the biosynthesis of nocardicin A discussed. The gene encoding this transferase was cloned from a sublibrary of Nocardia uniformis DNA. Translation gave a protein of deduced mass 32,386 Da which showed weak homology to small molecule methyltransferases. However, three correctly disposed signature motifs characteristic of these enzymes were observed.

Amino Acid Sequence↗

PubMed1989

The fate map of the chick forelimb-bud and its bearing on hypothesized developmental control mechanisms.

Carbon particles and isotopic quail grafts were used as markers to study the salient features of the fate map of the chick forelimb between stages 20 and 27. The grafting technique confirmed the reliability of the carbon method: they both revealed striking asymmetries in which apical mesodermal tissue was progressively displaced in a proximal direction (as would be expected on the basis of growth by net apical addition of tissue) but also in a preaxial direction, while postaxial tissue became elongated in the direction of limb outgrowth. Ectoderm showed a similar preaxial-postaxial asymmetry but became displaced from initially underlying mesoderm. In marked contrast to mesoderm, distal ectoderm remained at a constant distance from the apical ectodermal ridge (or became incorporated into it), thus implying that the ectodermal sheet is anchored distally and grows by uniform stretching proximally. Within the ectoderm itself, the outer peridermal layer is displaced distally relative to the underlying epidermal basal layer. Peripheral mesoderm showed patterns of displacement which were intermediate between those of ectoderm and chondrogenic core mesoderm. It is argued that such morphogenetic phenomena are integral components of developmental mechanisms of significance in the control of pattern generation. Implications of the interpretation and use of the fate map in relation to theories of limb development, particularly those based on mechanisms defined in terms of limb axes, are reviewed.

Animals↗

PubMed1980

Transit of a meal through the stomach, small intestine, and colon in normal subjects and its role in the pathogenesis of diarrhea.

A method for measuring the transit time of a meal, containing sausages, mashed potato, baked beans, and a pineapple custard dessert, through the gastrointestinal tract was evaluated in 14 healthy volunteers. Gastric emptying was determined by incorporating a radioactive marker in the meal and counting over the surface of the stomach using a crystal scintillation detector. Small intestinal transit time was determined by measuring breath hydrogen excretion and by estimating the radioactivity over the cecum. Finally, whole gut transit time was measured by incorporating radiopaque plastic markers or carmine red in the meal and estimating the appearance of these markers in the stool. Our results showed that measurements of small intestinal transit time were reproducible and in the majority of subjects the increase in hydrogen excretion occurred at the same time as the increase in radioactive counts over the surface of the cecum. The passage of the first marker in the stool coincided with the appearance of carmine red. There were no significant correlations between small intestinal transit time and whole gut transit time or the half time for gastric emptying. Incorporation of 10, 25, and 40 g lactulose into our standard meal in place of sucrose increased the rate of transit through the small intestine but did not significantly alter the rate of gastric emptying or the whole gut transit time. Total stool weight for 48 hr after ingestion of the meal was inversely related to whole gut transit time but not to small intestinal transit time suggesting that the tendency to develop diarrhea in response to a meal containing unabsorbable carbohydrate depends more on the lack of colonic accommodation than on the rate of small intestinal transit. Finally, there was no significant correlation between the measurements of small intestinal transit time after a drink of lactulose and the transit time of a meal in the same subjects.

Adult↗

PubMed1974