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Biomedical subjects

R R Tice

Publications and source records attributed to R R Tice.

81 records · Page 5Linked to original sources
PubMed1977

In vivo analysis of cellular replication.

The number of previous cell replications that a metaphase cell has undergone in the presence of BrdUrd can be determined by the differential fluorescent patterns of metaphse chromosomes stained with Hoechst dye 33258. To examine if this technique could be applied to analyzing cell cycle kinetics in vivo, we infused Wistar rats with BrdUrd for 7.5-33 hr at concentrations of the nucleotide analog that did not inhibit cellular replication. Examination of the frequency of one, two, and three or more replication cycle cells as a function of BrdUrd infusion time indicates that cell replication times for rat bone marrow cells are relatively homogeneous. Analysis of this data with a computer simulation model produced a mean cell cycle duration of 9.2 hr, which is compatible with the fastest times obtained with radioisotope studies. These results support the potential of nonradioisotope analysis of cell replication in vivo.

Animals↗
PubMedDate not supplied

Aneuploidy in mammalian somatic cells in vivo.

Aneuploidy is an important potential source of human disease and of reproductive failure. Nevertheless, the ability of chemical agents to induce aneuploidy has been investigated only sporadically in intact (whole-animal) mammalian systems. A search of the available literature from the EMCT Aneuploidy File (for years 1970-1983) provided 112 papers that dealt with aneuploidy in mammalian somatic cells in vivo. 59 of these papers did not meet minimal criteria for analysis and were rejected from subsequent review. Of the remaining 53 papers that dealt with aneuploidy induction by chemical agents in mammalian somatic cells in vivo, only 3 (6%) contained data that were considered to be supported conclusively by adequate study designs, execution, and reporting. These 3 papers dealt with 2 chemicals, one of which, mercury, was negative for aneuploidy induction in humans, and the other, pyrimethamine, was positive in an experimental rodent study. The majority of papers (94%) were considered inconclusive for a variety of reasons. The most common reasons for calling a study inconclusive were (a) combining data on hyperploidy with those on hypoploidy and/or polyploidy, (b) an inadequate or unspecified number of animals and/or cells per animal scored per treatment group, and (c) poor data presentation such that animal-to-animal variability could not be assessed. Suggestions for protocol development are made, and the future directions of research into aneuploidy induction are discussed.

Aneuploidy↗
PubMedDate not supplied

The induction of micronucleated polychromatic erythrocytes in mice using single and multiple treatments.

Studies are reviewed in which the effect of treatment/sample protocol on the induction of micronucleated polychromatic erythrocytes (MN-PCE) in male B6C3F1 mice by 3 carcinogens (benzidine, dimethylbenzanthracene and mitomycin C) were evaluated. 3 different treatment/sampling protocols were used, involving from 1 to 3 consecutive daily treatments and from 3 to 1, respectively, consecutive daily samplings beginning 24 h after the last injection. The results indicate that the 3-day injection/single sample time protocol eliminates the need for multiple sample times, minimizes the number of animals required in a study, decreases the time needed for data collection and simplifies data analysis. A comparison of the frequency of induced MN-PCE in peripheral blood and bone marrow suggests that, following a 3-injection protocol, either tissue can be used with equal efficiency.

9,10-Dimethyl-1,2-benzanthracene↗
PubMedDate not supplied

The use of bromodeoxyuridine labeling in the human lymphocyte HGPRT somatic mutation assay.

The autoradiographic assay developed by Strauss and Albertini (1979) to quantitate human in vivo somatic mutation at the hypoxanthine guanine phosphoribosyl-transferase locus uses tritiated thymidine to identify mutant cells by their ability to pass through 'S' phase in the presence of 6-thioguanine. An alternative method, based on the incorporation of bromodeoxyuridine (BrdUrd) into the DNA of proliferative cells, followed by differential staining with the fluorescence-plus-Giemsa method, was used to identify 3 classes of lymphocyte nuclei: (a) small darkly stained nuclei, (b) large, reddish-colored nuclei with an apparent nucleolus, and (c) large, bluish-colored nuclei. By double labeling with BrdUrd and tritiated thymidine, it was determined that only the nuclei of the third class had incorporated BrdUrd. These results demonstrate that the technique used for sister-chromatid differentiation can be used to detect putative HGPRT mutants and to determine variant frequencies at the HGPRT locus.

Autoradiography↗
PubMedDate not supplied

DNA damage and repair with age in individual human lymphocytes.

Previous biochemical studies on DNA repair competence and aging have been limited to techniques, such as alkaline elution or nucleoid sedimentation, involving mass cell populations. These techniques provide no information about the distribution of DNA damage and repair among individual cells and are unlikely to detect age-dependent changes affecting a minor fraction of the cell population. We have recently described a microgel electrophoretic assay (Singh et al., 1988) that measures, at the level of the individual cell, single-strand DNA breaks and alkali-sensitive sites. Here, we employ this method to analyze DNA damage and repair in lymphocytes isolated from the peripheral blood of 31 subjects (23 males and 8 females aged 25-91 years) and exposed in vitro to 200 rads of X-irradiation. While basal (pre-irradiation) levels of damage were independent of the age of the donor, an age-dependent increase in DNA damage was observed immediately following irradiation. For all subjects, the mean level of DNA damage was restored to pre-irradiation control levels within 2 h of incubation at 37 degrees C. However, a distribution analysis of DNA damage among cells within each sample indicated the presence of a few highly damaged cells (4-16%) in the 2-h sample, the occurrence of which was significantly more common among aged individuals. These data indicate an age-related decline in DNA repair competence among a small subpopulation of lymphocytes.

Adult↗