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The infection of chimpanzees with ECHO viruses.

The oral and parenteral infections of chimpanzees receiving echo Types 6 and 4 viruses successively are described. The two infections, spaced 2(1/2) months apart, and given by the same route in each animal, failed to induce overt disease. The inapparent infections were demonstrated by virus excretion in the throat and the stools and the development of neutralizing antibodies. Complement-fixing antibodies also appeared after Type 6 infection, but fell more rapidly than the neutralizing antibodies. After oral infection, echo-6 virus was found for equal periods in both the throat and feces, but echo-4 persisted in the throat for much longer periods than in the lower bowel. Almost no virus carriage occurred after parenteral inoculation. No true viremia was exhibited in any of the animals. One of the chimpanzees had neutralizing antibodies against Type 6 virus in its pre-inoculation serum. It responded extraordinarily to the Type 6 exposure, developing antibody levels of 1:50,000 to echo-6, of 1:1024 against the echo-6' variant, and of 1:64 against the echo 6'' variant. Although Type 4 antibodies developed after the exposure, they proved difficult to measure by ordinary methods. However, they could be satisfactorily assayed by the plaque reduction method. Three other chimpanzees fed echo-2, echo-3, and an untypable echo virus, respectively, yielded results confirming those established with Types 4 and 6.

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Studies of mouse polyoma virus infection. 1. Procedures for quantitation and detection of virus.

THREE PROCEDURES HAVE BEEN COMPARED FOR USEFULNESS IN TITRATION AND DETECTION OF POLYOMA VIRUS: production of cytopathic effect (CPE) in mouse embryo tissue culture, production of HI antibody after inoculation into weanling mice (MAP test), and production of tumors in suckling hamsters during a 3 to 5 week observation period. The tissue culture and mouse antibody production tests were generally comparable in sensitivity, reproducibility, and time required to obtain results. Titration by tumor production in suckling hamsters was not suitable for quantitation because of marked variation in susceptibility among animals. Virus was detected in tissues of normal mice from spontaneously infected colonies by either production of CPE in mouse embryo tissue culture or by the MAP test; virus was found in organs of 15 (58 per cent) of 26 mice with antibody, and 2 (8 per cent) of 24 mice without antibody.

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Studies on the Shope rabbit papilloma virus. II. The location of infective virus in papillomas of the cottontail rabbit.

A method has been devised to determine the location of infective Shope virus in the papillomas of cottontail rabbits. Frozen sections of the growths were burned selectively with a microcautery to destroy either the keratinized or proliferating layer and the sections were then applied directly to the sensitized epidermis of domestic rabbits. No papillomas appeared when the keratohyaline and keratinized areas had been eliminated leaving the proliferating cell layer, whereas papillomas arose when the proliferating cell areas were destroyed leaving the keratohyaline and keratinized layers. The results indicate that infective Shope papilloma virus is situated mainly, perhaps entirely, in the keratohyaline and keratinized areas of cottontail papillomas. This is in accord with the previous disclosure by the fluorescence technique that virus antigen in demonstrable quantity is present only in these situations.

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Factors determining pathogenicity of variants of ECHO 9 virus for newborn mice.

While some strains of ECHO 9 virus were found to be completely incapable of multiplying in newborn mice or even of being adsorbed by their tissues (e.g., the prototype Hill strain), other naturally occurring strains readily multiplied even after inoculation of as little as 3 TCD(50) of virus. With the multiplying strains, the infection remained clinically inapparent except after inoculation of very large doses, usually in the range of 10(5) to 10(7.5) TCD(50). Investigation of the question why such large doses were required to produce paralysis indicated that for paralysis to occur virus multiplication had to reach a level of 10(8) TCD(50) or more within 4 days after inoculation of mice less than 1 day old. The reason for this was found in the fact that at 5 to 6 days of age the mice lost their susceptibility to paralysis even when multiplication was capable of progressing to the indicated high level. Thus, speed of multiplication and extent of muscle involvement before the 5th day of life were the determining factors. Passage in tissue culture had no effect except to yield a larger dose for inoculation, while serial propagation in mice resulted in a gradual enrichment of virus particles capable of more rapid multiplication in mice and in a concurrent greater paralytogenic activity of smaller doses.

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Studies of mouse polyoma virus infection. V. Relation of virus infection to lymphocytic neoplasms of the mouse.

The relationship of polyoma (parotid tumor) virus infection to spontaneous and x-ray induced lymphocytic neoplasms (leukemia) in mice has been studied in two sublines of the high leukemic AKR strain, in the high leukemic C58 strain and in the x-ray responsive C3Hf/Bi strain. There was found to be no correlation of hemagglutination-inhibiting (HI) antibody with leukemia appearing at various times throughout life, and there was no evidence of increased risk of developing leukemia in mice with prior antibody to polyoma virus. Virus isolation experiments accomplished with AKR leukemic mice showed that HI antibody status of these mice had the same significance with regard to presence or absence of detectable virus as was previously observed for non-leukemic mice from infected colonies. The results of this study were interpreted as supporting the concept that the repeated association of polyoma virus and experimental transmission of leukemia is fortuitous.

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The vacuolating virus of monkeys. I. Isolation, growth characteristics, and inclusion body formation.

A vacuolating virus isolated from uninoculated patas monkey kidney cultures was found to be serologically identical with SV(40), a virus previously found in association with rhesus and cynomolgus monkeys. Detection of the patas virus was facilitated when patas cells were exposed to x-ray treatment. Rhesus monkey and human cells were relatively insusceptible to the virus, although it persisted in these cells for a long period of time. Distinct intranuclear inclusions were detected in infected patas cultures 2 to 3 days before cytoplasmic vacuoles were noticeable. Cultures previously infected with PA-57 virus did not affect yields of poliovirus, and doubly infected cells were easily distinguished in stained preparations.

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The vacuolating virus of monkeys. II. Virus morphology and intranuclear distribution with some histochemical observations.

Cells infected with the vacuolating virus, SV(40), respond by swelling to several times their normal volume. Within enlarged nuclei, virus-containing inclusions appear which are acidophilic and Feulgen-positive. The formation of nuclear inclusions is followed by the appearance of cytoplasmic vacuoles and then shrinkage of the cell. Inclusions were found to exhibit unique double staining when a light-green counterstain was used in the Feulgen reaction. The virus is of low electron density, round, and 300 A in diameter. It occurs in large numbers, singly and in short chains, and it appears to multiply at the expense of chromatin.

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An antiviral substance from Penicillium funiculosum. II. Effect of helenine upon infection in mice with Semliki Forest virus.

Helenine exerts a therapeutic effect against Semliki Forest virus infections of mice. Cures, that is to say the survival of treated animals, were more frequently observed in Semliki Forest virus infections than they were in SK virus infections. It is believed that this difference in end-result probably represented only a quantitative difference in the therapeutic effect of helenine against these two viruses and not a qualitative difference in its mechanism of therapeutic action. The findings reported in this paper with regard to the treatment of Semliki Forest virus infections with helenine parallel very closely those described in an accompanying paper which deals with the action of helenine on SK. virus infections.

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