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We have studied the reactions of hamster embryo cells transformed by ultraviolet-inactivated herpes simplex type 2 (333-8-9 T cells) to infections with adeno-associated satellite virus (AAV) and its adenovirus helpers. Resident HSV structural antigens were not detectable in early or late passage of 333-8-9 T cells. AAV structural antigens were not detected in these cells unless the cells were coinfected with a helper adenovirus. In early passage 333-8-9 T cells were permissive to infections with simian adenovirus SV15 whereas normal hamster cell line LSH was nonpermissive. In some late passages of 333-8-9 T cells infections with SV15 adenovirus led to the production of viruslike particles whose morphology was identical with reoviruses.

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Purified human rhinovirus type 14 has been subjected to acid and alkaline degradation and the effects of hydrogen ions on particle morphology and infectivity are being studied. Considerable morphological degradation of the capsid occurred by pH 5.5. Breakdown may be by loss of discrete capsomere groups accompanied by release of the viral genome in the form of a ribonucleoprotein strand. Examination of virions prepared for electron microscopy by the “critical point” procedure which results in essentially 3 dimensional specimens confirms this finding. Under alkaline conditions there is very little loss of infectivity up to pH 9.8. There is a precipitous loss in infectivity at around pH 10.9 accompanied by complete break up of viral capsid morphology.

Contour lengths, molecular weights, and topological parameters of single molecules of nucleic acids can be studied accurately by electron microscopy. However, specialized equipment and highly purified samples of DNA or RNA in an amount of 10-100 μgm have been required. We have developed a simple micro-diffusion technique which uses less than 0.01 (μgm of nucleic acid. Droplets containing the nucleic acid are placed on a clean teflon sheet. Cytochrome C powder is added by needle to form a monofilm. After 20-30 minutes diffusion time, there is usually enough material attached for the film to be picked up on a suitable specimen grid, shadow-cast or stained, and examined in the electron microscope. Each droplet provides material for a single observation, but experiments may be carried out contiguously on the sheet.The method has been developed for viral nucleic acids, but can also be applied to solutions of virus particles where the genomes can be released by osmotic shock or by other physical procedures. Less than 1010 particles per ml are required for this modification. Examples of nucleic acids liberated from papovaviruses, SV40, rabbit papilloma, human papilloma, and from adeno-associated satellite virus will be presented. Viral nucleic acids (DNAs from SV40 and adenovirus, RNA from bacteriophage R17) prepared by conventional techniques will also be shown.