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Biomedical subjects

B L Nicholson

Publications and source records attributed to B L Nicholson.

24 records · Page 2Linked to original sources

Inhibition of cellular DNA synthesis in cells infected with infectious pancreatic necrosis virus.

In asynchronous RTG-2 cell cultures infected with infectious pancreatic necrosis (IPN) virus, inhibition of cellular DNA synthesis, but not protein synthesis, was detected 5 to 6 h postinfection and was 80 to 90% complete by 7 to 8 h. Inhibition of DNA synthesis was largely abolished by UV irradiation of the virus. Sedimentation analyses of phenol-extracted DNA indicated that native cellular DNA was not degraded during infection. Sedimentation on alkaline sucrose gradients of DNA from cells pulsed with radioactive thymidine for varying periods indicated that elongation of nascent DNA chains proceeded normally in infected cells. These and previous results suggest that IPN virus infection results in a reduction of the number of chromosomal sites active in DNA synthesis but does not affect the rate of polymerization at active sites. Cells synchronized with excess thymidine and hydroxyurea and infected with virus at the time of release from the block demonstrated an inhibition of DNA synthesis 3 h postinfection. Cells infected 4 h prior to release continued to synthesize normal amounts of DNA for 1 to 2 h after release. These results indicated that DNA synthesis in early synthetic phase is relatively insensitive to inhibition by IPN virus.

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Immunofluorescent study of the replication of infectious pancreatic necrosis virus in trout and Atlantic salmon cell cultures.

Cell cultures of trout gonad tissue (RTG-2) and Atlantic salmon heart, kidney, liver, and spleen tissue were inoculated with 50 50% tissue culture infective doses (TCID(50)) of infectious pancreatic necrosis (IPN) virus per cell, and the titer of cell-associated and released virus was determined from 2 to 16 h postinoculation (PI). Cover slips were collected over the same period and stained for IPN viral antigen by the direct immunofluorescent (FA) technique. Viral replication was detected after a latent period of approximately 2 to 4 h and reached a peak titer of 10(8.2) to 10(8.4) TCID(50) per ml at 8 to 10 h PI. The release of virus was more rapid in Atlantic salmon cells than in RTG-2 cells. Viral antigen was first detected by FA from 3 to 4 h PI. Approximately 75 to 80% of the cells contained antigen in the cytoplasm 9 to 11 h PI. The direct FA technique was found to be a sensitive method for detecting IPN virus in infected cells. Three strains of IPN virus were tested for serological cross reactions by FA and virus neutralization tests.

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Host influence on the antigenic composition of the Kilham rat virus.

The Kilham rat virus (RV) was found to vary in infectivity and antigenic characteristics when propagated in two different host systems. Both cross hemagglutination-inhibition (HI) and cross virus-neutralization tests revealed that a nonreciprocal or one way cross exists between RV propagated in rat embryo cell cultures (RE-RV) and RV propagated in suckling hamsters (H-RV). Specifically, immune serum to RE-RV inhibited hemagglutination and infectivity by both viruses to the same extent. Immune serum to H-RV, however, exhibited higher HI and neutralization titers to H-RV than to RE-RV. Infectivity studies demonstrated that, although both viruses were able to infect either host, the virus showed a higher infectivity titer for the last host in which it had been propagated. Serological studies indicated that H-RV possesses an antigen(s) not found on the RE-RV. This host-controlled variation did not result after a single passage in the new host, but rather required at least three passages for a complete conversion to occur, and did not appear to result from the incorporation of unaltered host antigens into the virus particle. Solubilization of purified RV propagated in each host with sodium dodecyl sulfate, 2-mercaptoethanol, and urea followed by electrophoresis in polyacrylamide gels indicated that the number of components in the protein of each virus was not the same.

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