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Biomedical subjects

R W Simpson

Publications and source records attributed to R W Simpson.

At least 55 records · Page 3Linked to original sources
PubMed1977

Hormonal and metabolic responses to glucagon in diabetes mellitus.

The metabolic and hormonal responses to glucagon (1 mg, subcutaneous) were studied in twelve diabetic and twelve non-diabetic subjects. Diabetics showed a GH response, which although commencing slightly earlier, did not otherwise differ from that of the controls. There was the expected diminished insulin response to glucagon among the patients on oral agents. The GH response to glucagon in the diabetics began before there was any significant fall in blood glucose (BG). The hyperglycaemic response in the diabetics peaked later, was greater in amplitude and was more prolonged. In this group both pyruvate and lactate changes were delayed and diminished in amplitude. The diabetics showed a biphasic response of both acetoacetate and 3-hydroxybutyrate to glucagon; a small transient significant rise in the first 2 h followed by a second rise after 3h. In contrast the controls showed a significant fall in these metabolites during the initial 2 h, before a rise after 3 h. The significance of these hormonal and metabolic responses is discussed.

Acetoacetates↗
PubMed1976

Transcriptase activity and genome composition of defective influenza virus.

Eight genome RNA segments are present in both normal and von Magnustype influenza virus preparations and all species are transcribed by the virion-associated polymerase. Although the RNA polymerase activity and the amount of the three largest RNA segments are reduced in defective influenza virus preparations, these reductions do not appear to be great enough to account for the much greater loss of infectivity.

DNA-Directed RNA Polymerases↗
PubMed1975

Recovery of infectious proviral DNA from mammalian cells infected with respiratory syncytial virus.

The DNA fraction from a line of bovine embryonic kidney cells originally exposed as primary cultures several months earlier to a temperature-sensitive (ts) mutant of respiratory syncytial (RS) virus could be used to transfect human HEp-2 cells with the production of infectious RS virus. The DNA donor cells, designated BEK/RS ts, retained their healthy fibroblastic appearance during continuous cultivation at a temperature (39 degrees) restrictive for growth of the original infecting mutant and showed no evidence for RS virus replication or viral antigen synthesis when directly examined for these activities by conventional methods. The infectious property of the DNA from BEK/RS ts cells was abolished by exposure of the nucleic acid preparation to DNase (but not RNase) or by pretreatment of recipient HEp-2 cells with actinomycin D or mitomycin C. The latter drug treatments substantially enhanced the replication of infecting wild-type RS virus in HEp-2 cells. Viral isolates derived from the progeny of a DNA transfection included clones possessing several genetic markers of the RS ts mutant originally used to infect BEK/RS ts cells and other virus clones that appeared to be either hybrid or wild-type for phenotypic properties such as their temperature sensitivity. An infectious proviral DNA was also detected in a line of virogenic HEp-2 cells (HEp-2/RS) persistently infected with respiratory syncytial virus after exposure to the wild-type strain 2 years earlier.

Cell Line↗
PubMed1975

Virion-associated transcriptase activity of influenza recombinant and mutant strains.

A comparative study of the in vitro reaction kinetics of the virion RNA polymerase of influenza A strains WS and WSN was conducted to establish phenotypic differences for enzyme activity that might be exchanged as genetic markers among recombinants of these viruses. Characteristically, the RNA polymerase activity of WS virus showed an initial rate of synthesis about two- to threefold higher than that of WSN when assayed at 32 C. The two strains were also distinguishable by comparing the transcription rates of each strain at 32 and 37 C. The initial rate of WS was invariably higher at 37 than at 32 C, whereas the opposite was found with WSN. When a series of recombinants obtained from mixed infections with the WS and WSN viruses were examined for virion transcriptase activity, it was found that the two polymerase related markers behaved as properties which segregated independently of each other and of additional nonselective markers that were scored. Seven temperature-sensitive mutants of WSN virus representing distinct recombination-complementation groups were found to show a diminished transcriptase activity as compared to wild-type virus, and one of these clones (ts 24) was largely deficient for this function. None of these mutants appeared to possess a heat-liable virion polymerase.

Cells, Cultured↗
PubMed1972

Transcription of the influenza ribonucleic acid genome by a virion polymerase. 3. Completeness of the transcription process.

The virion ribonucleic acid (RNA) polymerase of influenza strain A(0)/WS transcribed at least 81% of the viral genome in vitro. The polymerase is shown to be associated with each of the major size classes of the virion RNA-ribonucleoprotein complexes. Under optimal in vitro conditions, at least 45% of the RNA contained in a population of influenza virions was involved in a repetitive transcription process. The detectable proteins associated with enzymatically active complexes containing RNA, ribonucleoprotein, and polymerase have been identified by polyacrylamide gel electrophoresis.

Centrifugation, Density Gradient↗
PubMed1971

RNA-dependent RNA polymerase activity associated with virions and subviral particles of myxoviruses.

In vitro RNA synthesis is catalyzed by an enzyme found in purified virions of each of nine influenza strains tested. The extent of [(3)H]UTP incorporation observed is strain dependent, with an obligate requirement for Mn(2+) ions and RNA template. At least 30% of purified product RNA shows resistance to degradation by ribonuclease A and T1 before and after annealing with exogenous RNA from influenza virions.

Catalysis↗
PubMed1971

Transcription of the influenza ribonucleic acid genome by a virion polymerase. I. Optimal conditions for in vitro activity of the ribonucleic acid-dependent ribonucleic acid polymerase.

In vitro reaction conditions have been determined for the maximal synthesis of product ribonucleic acid by the influenza (WSN) virion ribonucleic acid polymerase. The reaction requires the presence of all four triphosphates, Mg(2+) and Mn(2+) ions, monovalent cations, nonionic detergent, and ribonucleoside triphosphates at concentrations above certain threshold values. The optimum pH for the reaction is around 8.0 to 8.2 and the kinetics of product synthesis are linear through at least 6 hr when incubated at 31 to 33 C.

Animals↗
PubMed1971

Transcription of the influenza ribonucleic acid genome by a virion polymerase. II. Nature of the in vitro polymerase product.

The properties of the ribonucleic acid (RNA) synthesized by the influenza (WSN) virion polymerase have been investigated. Most of the product RNA is synthesized in association with virion RNA species from which it can be released by heat treatment as single-stranded, ribonuclease-sensitive polynucleotides (average molecular weight, 2-hr sample, about 10(5) daltons). At least 95% of the product is complementary to the viral RNA species. On the basis of the molar ratio of the RNA species isolated from a (3)H-uridine-labeled virus preparation, it was calculated that the WSN genome consists of seven pieces of RNA with a sum molecular weight of about 5 x 10(6) daltons.

Animals↗
PubMed1971

Deoxyribonucleic acid polymerase of Rous sarcoma virus: reaction conditions and analysis of the reaction product nucleic acids.

Reaction conditions for Rous sarcoma virus ribonucleic acid (RNA)-instructed deoxyribonucleic acid (DNA) polymerase activity are described whereby the viral RNA is relatively protected from endogenous or added nuclease activity. Three analyses of reaction product nucleic acids ((3)H-RNA, (32)P-DNA) were compared, namely, gel electrophoresis, Cs(2)SO(4) gradient centrifugation, and hydroxyapatite column chromatography. It was found that hydroxyapatite analysis could be misleading unless the state of the template RNA was monitored concomitantly with the DNA analysis. Gel electrophoresis and Cs(2)SO(4) gradient centrifugation gave comparable results. It was concluded that analyses of the product of reverse transcriptase reactions should not only refer to the template RNA and product DNA species, but also be performed with virus or viral RNA which do not have or obtain nicks in the 60S RNA. Otherwise, interpretation of the results would have the ambiguity of potential artifacts caused by those degraded RNA molecules.

Animals↗