Occurrence of free, defective viral DNA in a hamster tumor induced by human papovavirus BK

We characterized viral DNA present in an osteosarcoma cell line (Os-513), which had been originally induced in a hamster with human papovavirus BK (BKV), by hybridization of ³²P-labeled BKV DNA to DNA extracted from cells, undigested or digested with various restriction endonucleases, fractionated by electrophoresis, and transferred to a nitrocellulose filter. Os-513 was found to contain free viral DNA and high molecular weight DNA with viral sequences. The free viral DNA seemed to be defective, for its size was about two-thirds of the wild type. The deletion, one-third of the entire BKV DNA, extended over a segment between map positions 0.72 and 0 which includes the leader and intervening sequences and part of the body sequences for late mRNAs. The high molecular weight DNA with viral sequences appeared to be composed of tandem, head-to-tail repeats of the same defective BKV DNA as the free DNA.     

Genome rearrangements of bovine rotavirus after serial passage at high multiplicity of infection

After serial passage at high multiplicity of infection of standard bovine rotavirus in MA104 cells, different genome rearrangements occurred in which segment 5 was lost from the RNA profile and distinct additional bands of double-stranded (ds) RNA were found in positions on gels between segments 1 and 6. It was shown that some of the additional RNA bands contained segment 5-specific sequences. The additional RNA bands were transcribed in vitro to apparent full length. Analysis of the proteins synthesized in cells infected with viruses possessing rearranged genomes showed that in all cases the product of RNA segment 5, VP5, was missing; however, in one case an abnormal protein was observed which corresponded in size to the coding capacity of the mRNA transcribed from the additional genomic RNA band. Viruses with rearranged genomes could be plaque purified, and they grew in the absence of standard virus to titers comparable to those obtained from standard virus. In mixed infections of standard virus and virus possessing genome rearrangements, standard virus overgrew during passage at low multiplicity of infection whereas virus possessing genome rearrangements overgrew during passage at high multiplicity of infection.     

Characterization of two abundant mRNAs of Autographa californica nuclear polyhedrosis virus present late in infection

Cytoplasmic poly(A)(+) RNA isolated from Spodoptera frugiperda cells late after infection with Autographa californica nuclear polyhedrosis virus (30-40 hr pi) was fractionated according to size on denaturing methyl mercury gels. Two major RNA species (1.4 kb and 0.75 kb) and several minor RNA species were detected by ethidium bromide staining. The predominant RNA species of about 1.4 kb was considered to be polyhedrin mRNA because (1) in vitro translation of the RNA, which was eluted from methyl mercury gels, yielded a polypeptide of MW 33K, which comigrated with polyhedrin. (2) When poly(A)+ RNA was fractionated on a sucrose column and then translated in vitro, the distribution and abundance profiles of a 33K polypeptide product and of 1.4-kb RNA were similar. (3) The 33K polypeptide made in vitro and purified polyhedrin gave rise to similar patterns of peptides when digested with S. aureus V8 protease. The polyhedrin mRNA (1.4 kb) hybridized to BamHI-F and HindIII-V AcNPV DNA fragments and hybridization selection with BamHI-F AcNPV DNA yielded a 33K polypeptide, which comigrated with polyhedrin. The second RNA species (0.75 kb in size) hybridized to overlapping EcoRI-P and HindIII-Q regions of the AcNPV genome and translated into a methionine deficient polypeptide of MW = 8K. It was synthesized in large quantities late in the infection and appeared to be coordinately expressed with polyhedrin in infected cells. The 8K polypeptide was detected as early as 15 hr pi and was still synthesized at 60 hr pi.     

Psychosis as a predictor of response to lithium maintenance treatment in bipolar affective disorder

Sixty-six bipolar I lithium clinic patients were studied for a history of psychotic symptoms at some time during the course of their illness. Agreement between different sources of information was calculated, and the patient population was divided into psychotic and non-psychotic subgroups. Probability of remaining well on lithium for the different subgroups was analyzed by the life table method. Psychosis during mania appeared to be associated with especially good early lithium prophylaxis.     

Persistence and replication of the human cytomegalovirus genome in lymphoblastoid cells of B and T origin.

Human lymphoblastoid cells infected in vitro with human cytomegalovirus (CMV) were examined by DNA-DNA reassociation kinetics at various times after virus infection. The results indicate that CMV DNA replicates in two B lymphoblastoid cell lines (Raji and Simpson) and one T-cell line (Molt-4). Multiple copies of the entire CMV genome were found in each cell line. After an initial round of CMV DNA replication, the number of CMV DNA copies per cell gradually decreased with time in Raji and Molt-4 cells. In contrast, Simpson cells maintained a fairly constant number of CMV DNA copies throughout the 15-day experiments.     

Inhibition of vaccinia virus late protein synthesis by isatin-beta-thiosemicarbazone: characterization and in vitro translation of viral mRNA.

Thespecific effect of istin-βthiosemicarbozone (IBT) was manifested after vaccinia virus late protein synthesis had commenced. At 6 hr after infection, viral protein synthesis was inhibited by about 9596. We confirmed that λ portion of the virus-specific RNA appears to be degraded (B. Woodson and W. K. Joklik, 1965, Proc. Nat. Acad. Sci. USA 54,946–953). Nevertheless, the amount of viral RNA that was capped, properly methylated, and polyadenylylated, was reduced by only about 50%. Moreover, RNA from IBT-treated cells stimulated cell-free protein synthesis to one-half the level obtained with RNA from control cells. Polyacrylamide gel electrophoretic analysis further demonstrated that RNA from IBT-treated cells was translated into late viral proteins in vitro. Thus, it seems possible that the inhibition of protein synthesis in IBT-treated cells does not result entirely or directly from either an inhibition of mRNA synthesis or from λ depletion of mRNA caused by accelerated degradation. An alternative possibility, that accelerated degradation is secondary to λ more immediate effect of the drug on protein synthesis, was considered.     

Cell division requirement for activation of murine leukemia virus in cell culture by irradiation.

Actively dividing cultures of AKR mouse cells were exposed to relatively low dose-rates of gamma radiation and tested for activation of endogenous leukemia viruses. Efficient and reproducible induction of virus was obtained with actively dividing cells, but cultures deprived of serum to inhibit cell division before and during gamma irradiation were not activated, even when medium with serum was added immediately after irradiation. These results show that cell division was required for virus induction but that a stable intermediate similar to the state induced by halogenated pyrimidines was not formed. In actively dividing AKR cell cultures, virus activation appeared to be proportional to the dose of gamma radiation; the estimated frequency of activation was 1.8 × 10^?5 per exposed cell and the efficiency of activation was approximately 0.012 inductions per cell per rad. Other normal primary and established mouse cell cultures tested were not activated by gamma radiation. The requirement of cell division for radiation and chemical activation may reflect some common mechanism for initiation of virus expression.     

Some studies on the mechanism of action of antibodies to nerve growth factor.

The effects of antibodies to the nerve growth factor from mouse salivary gland were examined in vitro and in vivo. Treatment of explants of receptive ganglia with antibody and complement did not produce cell damage as judged by the ability of the tissue to respond to nerve growth factor. New-born mice experimentally depleted of or genetically deficient in key complement components were susceptible to the action of the antiserum.These results show that the effect of the antibody is independent of complement and are consistent with the view that it acts by neutralization of endogenous nerve growth factor.