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Biomedical subjects

K Edwards

Publications and source records attributed to K Edwards.

At least 109 records · Page 6Linked to original sources

Studies on the organisation of the chicken genome and its expression during myogenesis in vitro.

DNA from the chicken genome was analysed both by isopycnic centrifugation in cesium salt density gradients and by reassociation analysis using hydroxyapatite (HAP) chromatography. Centrifugation in neutral CsCl revealed a single non-Gaussian band skewed toward the heavy side, but no discrete satellite components. In heavy metal (Ag+ or Hg++)-Cs2SO4 gradients, 4-8 satellite bands were revealed, comprising 5-9% of the total DNA. Purification of the satellites and recentrifugation in neutral CsCl demonstrated that 80-90% of this DNA would band in the shoulder, with the remainder in the main band. These satellites can account at most for 30% of the heavy shoulder DNA, thus most of the heavy shoulder DNA must be of lower repetition frequencies. Reassociation analyses of chicken DNA demonstrated that the complexity of the non-repetitive DNA is 9.49 X 10(8) nucleotide pairs, equivalent to about 90% of the haploid genome. Repetitive DNA comprises only 8-10% of the genome and has the following composition, relative to total DNA: 3.7% intermediate repetitive, 1.9% highly repetitive, and 3.9% "zero-time binding" DNA. This unusually low repetitive DNA content may be related to the small genome size of chickens, relative to other vertebrates, and to the presence of many microchromosomes in the chicken karyotype. Total cell RNA extracted from perfusion myoblasts, post-fusion myotubes, and myoblasts grown in BrdU was incubated in large excess with 3H-TdR labelled non-repetitive DNA and the resulting hybrids assayed by HAP chromatography. The amount of non-repetitive DNA represented in the RNA was found to increase from 7-8% in the myoblast stage to 10-11% in myotubes. An even smaller proportion, about 5%, is represented in the RNA of myoblasts prevented from differentiating by growth in BrdU.

Animals

Synthesis of albumin via a precursor protein in cell suspensions from rat liver.

The mechanism of the biosynthesis of albumin was studied in cell suspensions from rat liver. The cells were prepared by continuous perfusion of the liver in situ with 0.05% collagenase and 0.10% hyaluronidase and incubated under conditions optimized for the incorporation of amino acids into protein. Seven minutes after starting the incubation L-[1-14C]leucine was added, followed after 25 min by a 15 or 30-min chase with an 830-fold excess of non-radioactive L-leucine. Total protein, an albumin-like protein, and albumin were isolated from samples withdrawn immediately of total protein was found to remain constant after addition of the non-radioactive L-leucine, whereas that of the albumin-like protein decreased and that of albumin increased with incubation time. The increase in albumin radioactivity accounted for the decrease in radioactivity of the albumin-like protein, suggesting that the latter is a precursor of albumin. The precursor protein differed from albumin by an oligopeptide extension at the N-terminal end.

Albumins

Biosynthesis of albumin via a precursor protein in Morris hepatoma 5123tc.

The mechanism of albumin biosynthesis was studied in Morris hepatoma 5123tc in vivo and in hepatoma cell suspensions obtained by solubilizing the intercellular matrix with collagenase and hyaluronidase. In the in vivo experiments, L-[-14C]leucine was injected i.v. into rats bearing hepatomas in the muscles of both hind legs. After 14 min, tumors were removed and homogenized. A protein fraction quantitatively precipitable with antialbumin was isolated from the homogenate by acetone fractionation and precipitation with antiserum against serum albumin. This protein fraction was not homogeneous. With the use of 3 consecutive chromatographies on diethylaminoethyl cellulose, a very highly radioactive albumin-like protein could be separated from a large amount of only slightly radioactive albumin. In hepatoma cell suspensions incubated with L-[1-14C]leucine followed by a chase with excess nonradioactive L-leucine, radioactivity was incorporated first into the albumin-like protein and transferred thereafter into albumin, suggesting that albumin was synthesized via the albuminlike protein as precursor. In vivo, 1.8% of newly synthesized hepatoma protein was albumin or its precursor, compared with 1.2% in cell suspensions.

Albumins

The effect of triazolam on the sleep of insomniacs.

The effects of three oral doses of bedtime triazolam (0.25 mg, 0.5 mg, and 1.0 mg) a new benzodiazepine, on the laboratory sleep of insomniacs were studied in a double blind design which used the following 14 consecutive night schedule: 1-4 placebo; 5-11 drug; 12-14 placebo. Effects on sleep were measured objectively by conventional EEG/EOG/EMG sleep recordings and subjectively by questionnaires administered each morning. Side or toxic effects were assessed by screening physicals and questionnaires administered each morning and each evening and by a comparison of the prestudy vs. end-study physical exams and clinical lab tests. At the 0.5 mg dose triazolam significantly reduced several objective and subjective measures of insomnia. It had lesser effects at the 0.25 mg dose and equal or greater effects at 1.0 mg dose. There were no remarkable side or toxic effects at any dose.

Adult