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Biomedical subjects

R A Daw

Publications and source records attributed to R A Daw.

7 recordsLinked to original sources
PubMed1991

Polymerase chain reaction for detection of human cytomegalovirus infection in a blood donor population.

We have used the polymerase chain reaction (PCR) to analyse 420 normal donor blood samples taken at a city centre donation site. Three sets of human cytomegalovirus (HCMV) primers specific to the HXLF6, immediate early and late antigen gp64 genes, of the alpha, beta and gamma antigen coding regions respectively, were used to allow for the possibility of sequence variation. There was perfect correlation between the three sets of primers. Latex agglutination and enzyme-linked immunosorbent assay (ELISA) were also employed to provide data for a comparative study. The complete data show that infection with human cytomegalovirus is not only age related but is also sex related. The female population examined using PCR reached a peak infection rate of 80% by the age of 40-50 years whereas the male population reached a 98% infection rate following an almost linear increase with age. Latex agglutination data shows a similar picture although the infection rate peaks around 20% lower than with PCR. The data shows an increase in sensitivity using the PCR rather than the serology although the clinical significance of this has yet to be determined. The work presented here also demonstrates the suitability of the polymerase chain reaction as a potential diagnostic and epidemiological tool.

Age Factors↗
PubMed1983

Immunoaffinity purification of human complement component C9 using monoclonal antibodies.

Five monoclonal antibodies to human C9 were produced. Three of the antibodies had affinities from 1.6 to 3.1 X 10(9) M-1 and 2 were of lower affinity, 0.1 to 0.3 X 10(9) M-1. Four distinct antigenic sites on C9 were detected. The lowest affinity antibody was immobilised on Sepharose 4B and used to isolate C9 from human plasma. A 1200-fold purification was obtained with yield of 30-40%. The purified antigen maintained full cytolytic activity.

Animals↗
PubMed1981

Direct measurement of the increase in intracellular free calcium ion concentration in response to the action of complement.

1. The effect of rabbit anti-(pigeon erythrocyte) antibodies plus human complement on the concentration of intracellular free Ca2+ in sealed pigeon erythrocyte 'ghosts' was investigated with the photoprotein obelin. 2. The addition of human serum, as a source of complement, to 'ghosts' coated with antibody caused a rapid increase in intracellular free Ca2+ after a lag of 20-40 s, as detected by an increase in obelin luminescence. 3. The increase in obelin luminescence could not be explained by release of obelin into the medium. It was also Ca2+-dependent in that extracellular EGTA abolished the effect and intracellular EGTA inhibited it and required the complete terminal complex (C56789). No effect was seen with C5678. 4. The concentration of intracellular free Ca2+ before addition of complement was approx. 0.3 microM. This increased to a maximum of 5-30 microM after complement addition and then remained constant for at least 1-2 min. 5. Antibody plus complement induced a rapid increase in 42K+ efflux and an inhibition of cyclic AMP formation. 6. When partially purified complement components (C5b-9) were used in 'reactive lysis' it was possible to inhibit the release of macromolecules from pigeon erythrocyte 'ghosts' by extracellular EGTA. 7. It was concluded that the increase in intracellular free Ca2+ concentration caused by anti-cell antibody plus complement occurred before cell lysis and may be involved in the mechanism of complement-induced cell injury.

Animals↗