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Biomedical subjects

C C Randall

Publications and source records attributed to C C Randall.

At least 19 recordsLinked to original sources

Induction of recurring myositis ossificans by intrathymic injection of 3-methylcholanthrene-treated cells in rats.

Studies were performed to ascertain the effects of transplantation of thymic cells exposed in vivo to 3-methylcholanthrene (3-MC) on the induction of malignancies in Copenhagen rats. Three recipient rats unexpectedly developed tumors which bore histological resemblance to myositis ossificans of humans. Specifically, histology revealed areas of peripheral ossification with the appearance of zones of primitive osteoid with a central cellular area. Other areas of the lesions were less well organized into characteristic zones or were more or less heterogeneous. The primary, as well as recurring, lesions appeared in the axilla and were well circumscribed, 24-68 g in weight and 2-7 cm in diameter. Flow cytometric analyses of DNA content indicated that these tumors contained cells with abnormal DNA characteristics as well as proliferating cells. Coupled with the observation that after excision these tumors recurred, the data suggest that these myositis ossificans lesions were malignancies.

Animals↗

Thymic tumors induced by 3-methylcholanthrene treatment of rat Peyer's patches.

Studies were performed to characterize thymic tumors which were induced after a single injection of 500 microgram or 3-methylcholanthrene (3-MC) into surgically exposed Peyer's patches (PP) of Copenhagen rats. Detailed gross, histological, and morphological analyses revealed thymic tumors differing in size and weight (1 to greater than 8 g) with distorted architecture and infiltration by lymphocytes and epithelial cells in varying proportions. Approximately 25% of the rats with thymic tumors exhibited abnormal spleens, whereas 66% developed low grade leukemias. A majority of the thymic tumors contained cells which exhibited (1) phenotypic markers characteristic of normal thymocytes, (2) abnormal DNA, and (3) increased percentages in S + G2 phases of the cell cycle. Further studies of tumor cell isolates demonstrated an increased frequency of colony formation on soft agar, as well as the ability to elicit thymic tumors upon transplantation. Collectively these studies describe chemically induced thymic lymphomas.

Animals↗

Biological properties of equine herpesvirus type 1 DNA: transfectivity and transforming capacity.

DNA extracted from purified virions of equine herpesvirus type 1 (EHV-1) was examined for its transfectivity and transforming ability. The infectivity of the herpesvirus DNA was demonstrated by addition of calcium phosphate-DNA coprecipitates to monolayers of permissive horse cells, with resultant plaque formation. The efficiency of transfection (50 to 100 plaque-forming units/microgram of DNA) was reduced by treatment of the viral DNA with deoxyribonuclease or sonication but not with Pronase or antivirus neutralizing serum. When nonpermissive mouse 3T3 Cells lacking the enzyme thymidine kinase (TK-) were transfected with intact EHV-1 DNA, clones of cells transformed to the TK+ phenotype were isolated in selective HAT medium (hypoxanthine, aminopterin, thymidine), which prevents growth of the TK- parental phenotype. The efficiency of transformation ranged from one to five transformants per microgram of EHV-1 DNA. The TK activity of the biochemically transformed cells was characterized by biochemical, electrophoretic, and immunological techniques. By these criteria, the TK activity was identical to the EHV-1 TK and different from the host wild-type enzyme. In contrast to the parental TK+ 3T3 cells, the EHV-1-transformed TK+ cells were unable to grow in the presence of arabinosylthymine, a drug selectively phosphorylated by herpesvirus TKs. These results indicate that stable transfer of EHV-1 genes into nonpermissive cells can be achieved with purified viral DNA.

Arabinonucleosides↗

Biochemical transformation of deoxythymidine kinase-deficient mouse cells with UV-irradiated equine herpesvirus type 1.

A line of 3T3 mouse cells lacking deoxythymidine kinase (dTK-) was stably transformed to the dTK+ phenotype after exposure to UV-irradiated equine herpesvirus type 1 (EHV-1). Biochemical transformants were isolated in a system selective for the dTK+ phenotype (Eagle minimal essential medium containing 10(-4) M hypoxanthine, 6 X 10(-7) M aminopterin, and 2 X 10(-5) M deoxythymidine). Transformation was accompanied by the acquisition of a dTK activity with immunological, electrophoretic, and biochemical characteristics identical to those of the dTK induced by EHV-1 during productive infection. The transformed cells have been maintained in selective culture medium for more than 50 passages and have retained the capacity to express EHV-1--specific antigens. Spontaneous release of infectious virus has not been detected in the transformed lines, and the the cells were not oncogenic for athymic nude mice. In contrast to normal dTk+ 3T3 cells, EHV-1 transformants were unable to grow in the presence of arabinosylthymine, a drug selectively phosphorylated by herpesvirus-coded dTK's. These results indicate that a portion of the EHV-1 genome is able to persist in the transformed cells for many generations and be expressed as an enzymatically active viral gene product.

Animals↗

Replication of equine herpesvirus type 1 and type 3: resistance to hydroxyurea and thymidine.

The replication of equine herpesvirus type 1 (EHV-1) and type 3 (EHV-3) was unimpeded in three different cell types-equine epithelial cells, equine fibroblasts, and mouse fibroblasts-which had been blocked in their capacity to synthesize host DNA by 2.5 mM hydroxyurea (HU) or 2 mM thymidine (TdR). The rate of DNA synthesis in uninfected or equine herpesvirus-infected cells in the presence of HU or TdR was measured by pulse-labeling cell samples with a labeled DNA precursor at different times after infection. DNA synthesis in uninfected cultures was completely inhibited by both compounds. However, in cells infected with EHV-1 or EHV-3 and incubated in the presence of HU or TdR, a burst of DNA synthesis occurred which coincided in time with that of virus-specific DNA replication in infected cells without inhibitors. Analysis by cesium chloride isopycnic centrifugation confirmed that the DNA made in drug-treated, infected cells was viral. A possible mechanism of the HU and TdR resistance of the equine herpesviruses is the induction of a ribonucleotide reductase which is insensitive to inhibition by these compounds.

Animals↗

Odd-and even-numbered fatty acids. Their contrasting behavior in normal and fowlpox virus-infected epithelium.

Upon infection with fowlpox virus, the amount of odd-numbered fatty acids in chick scalp epithelium shows a significant decrease compared with control values. This effect begins quite early and progresses throughout the period of infection. Individual members of the odd-numbered family (C15--C27 inclusive) were quantitatively related to the group as a whole during most of the infection. Experiments involving the administration of labeled acetate in vivo demonstrated an increase in the synthesis of even-numbered fatty acids and a decrease in the synthesis of odd-numbered fatty acids in infected epithelium. The reduced synthesis of odd-numbered fatty acids in infected epithelium could also be demonstrated with labeled propionate. The influence of the alpha-oxidation pathway was assayed in chick scalp epithelium in vivo by the administration of [1-14C,9,10-3H] stearic acid. The C17 acids formed had a 3H/14C ratio similar to that of the C16 acids, indicating that most label incorporation into C17 was due to beta-oxidation to acetate followed by resynthesis into fatty acids. C17 fatty acids from control and infected epithelium had similar 3H/14C ratios, indicating that the alpha-oxidation pathway probably does not contribute to the differences in odd-numbered fatty acid content observed. In assays for fatty acid synthetase activty, both [14C] acetyl-CoA and [14C]-propionyl-CoA were used as initial acceptors. The specific activities of preparations from infected scalp were similar to those of control preparations with both substrates. These results suggest that there is no decline in the ability to utilize propionate for fatty acid synthesis in infected epithelium.

Acetates↗

Genetic relatedness of equine herpesvirus types 1 and 3.

The genetic relatedness of two types of equine herpesviruses (EHVs), 1 (EHV-1) and 3 (EHV-3), was determined by DNA-DNA reassociation kinetics. Denatured, labeled viral DNA probe was allowed to reassociate in the presence or absence of the second unlabeled viral DNA. The initial rate of reassociation of either labeled viral DNA was increased by the presence of the heterologous viral DNA to an extent indicating only 2 to 5% homology between the two EHV genomes. Moreover, labeled RNA extracted from EHV-3-infected cells hybridized to filter-immobilized EHV-1 DNA only 2 to 3 percent as efficiently as to the homologous EHV-3 DNA. These results demonstrate that the genital (EHV-3) and nongenital (EHV-1) types of EHVs exhibit very little genetic homology.

Base Sequence↗