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S A Latt

Publications and source records attributed to S A Latt.

At least 91 records · Page 5Linked to original sources
PubMed1981

Sister-chromatid exchanges: a report of the GENE-TOX program.

The effects of a number of chemicals on sister-chromatid exchange (SCE) frequencies in in vivo and in vitro systems are reviewed. Standardized protocols for future SCE testing in important systems, as well as for evaluation of test results, are presented. Data reported thus far suggest that SCE analysis may prove useful, especially at a secondary level, as a test of mutagenic carcinogens. Strengths and limitations of SCE analysis are summarized as a guide for future evaluation and use of this procedure.

Alkylating Agents↗
PubMed1981

Patterns of late replication in X chromosomes of human lymphoid cells.

Two predominant patterns of late X replication were observed in both short-term and established human lymphoid cultures. One pattern was found in a minority of short-term cultured T-cell metaphases, in most lectin-stimulated B cells, and, with minor variations, in established B-cell lines. In these cells, DNA replication terminated in the distal part of the long arm of the late X. A different pattern was found in the majority of lectin-simulated T cells and in the T-cell line CCRF-CEM. These cells exhibited terminal replication in a region of the long arm of the late X that was nearer to the centromere. It is speculated that the variations in replication patterns correlate with phenotypic and functional characteristics of human lymphoid subsets.

B-Lymphocytes↗
PubMed1981

Flow sorting of the mouse Cattanach X chromosome, T (X; 7) 1 Ct, in an active or inactive state.

Cells containing the mouse T(X;7)1Ct translocation were used to separate active and inactive X chromosomes. Cytophotometric data showed that the X(7) chromosome was at least 15% larger than any normal mouse chromosome. Cloned cell lines were established from male and female embryos with a normal karyotype and carrying the X(7) chromosome in an active or inactive state as shown by replication kinetics. As expected from the cytophotometric data, flow histograms of normal mouse chromosomes stained with 33258 Hoechst had five peaks. A sixth peak was present when the chromosomes came from mice carrying T1Ct. In this peak 6, 30 to 50% of the chromosomes were X(7) and there was almost no contamination by the normal X chromosome.

Animals↗
PubMed1980

Comparison of benzo(a)pyrene metabolism and sister chromatid exchange induction in mice.

Genetic differences in the inducible arylhydrocarbon hydroxylase (EC 1.14.14.2) (AHH) system, which is involved in the multi-step metabolism of hydrocarbons, are known to exist in both humans and mice. However, the predictive value of AHH activity in human or murine tissues, assayed as benzo(a)pyrene hydroxylation, as an index of individual susceptibility to mutagens and carcinogens, remains unclear because of apparent inconsistencies between results obtained from different in vitro and in vivo systems. This situation may in part reflect the complexity of the pathways involved in drug metabolism, which combine both activation and detoxification. To determine the relationship of metabolic potential to an easily quantified, short-term in vivo end point of genetic damage, we compared the ability of AHH inducible and uninducible mice to metabolize a procarcinogen, benzo(a)pyrene (BP), with the in vivo induction by BP of sister chromatid exchanges (SCEs). SCE induction has been shown to correlate with mutagenesis. We report here that although BP did cause an increase in SCEs in test animals, the extent of this increase did not differ between the inducible C57BL/6 mice and the uninducible DBA/2 mice. Moreover, prior exposure to an AHH inducer, 3-methyl-cholanthrene (3-MC), did not increase the number of BP-induced SCEs in C57BL/6 mice. This lack of correlation between benzo(a)pyrene hydroxylase (BP-OH) inducibility and SCE response reinforces the idea that other metabolic steps, such as detoxification or DNA repair, may influence the overall genetic impact of a drug.

Animals↗
PubMed1980

Induction of sister chromatid exchanges by BUdR is largely independent of the BUdR content of DNA.

The halogenated thymidine (dT) analogue, 5-bromodeoxy-uridine (BUdR), has a variety of effects on mammalian cells, including toxicity, suppression of differentiation, and mutagenesis. Although it is generally assumed that the effects of BUdR are due primarily to its presence in DNA, results from our laboratory have raised doubts about such assumptions. We have shown, for example, that BUdR mutagenesis in mammalian cells is determined by the concentration of BUdR in the medium rather than in DNA, and that mutagenesis can be suppressed by deoxycytidine (dC) without changing the amount of BUdR in DNA. BUdR has also been shown to induce sister chromatid exchanges (SCEs) in mammalian cells. Initial results suggested that the relationship between BUdR and SCEs might not be explained by a single factor, and various correlations between BUdR and SCEs have been proposed. However, the results to date have been inconclusive, because the experiments did not resolve as independent variables the concentration of BUdR in the medium and the amount of BUdR incorporated into nuclear DNA. We have now carried out experiments to resolve these two factors; the results indicate that the major factor in determining the frequency of SCEs is the concentration of BUdR in the medium.

Animals↗
PubMed1980

Interactions between pairs of DNA-binding dyes: results and implications of chromosome analysis.

A number of DNA-binding dyes, with spectral properties making them suitable as components of energy donor-acceptor pairs, are described. If such pairs are used to stain metaphase chromosomes, and if the energy acceptor (e.g., actinomycin D or methyl green) has a binding specificity opposite to the binding or fluorescence specificity of the donor (e.g., 33258 Hoechst, quinacrine or chromomycin A3), contrast in donor fluorescence can be enhanced, leading to patterns selectively highlighting standard or reverse chromosome bands or particular polymorphic regions. Such results presumably reflect chromosomal regions enriched in 10-20 base pair clusters to which the donor binds and fluoresces but to which the acceptor cannot bind. For other pairs, involving counterstains such as netropsin or echinomycin, which are not suitable as energy acceptors, specific changes observed in polymorphic region fluorescence are most likely due to binding competition between dyes. Dye pairs producing contrast by either method can be used to differentiate between homologous chromosomes or to facilitate detection of specific chromosomal rearrangements. Preliminary data indicate that contrast enhancement generated in fixed metaphase chromosomes spread on microscopic slides can also be observed in suspensions of unfixed metaphase chromosomes, reinforcing the expectation that the methodology described will be of use in flow cytometry.

Animals↗
PubMed1980

Energy transfer and binding competition between dyes used to enhance staining differentiation in metaphase chromosomes.

The ability of electronic energy transfer and direct binding competition between pairs of dyes to enhance contrast in human or bovine metaphase chromosome staining patterns is illustrated, and the relative effectiveness of these two mechanism compared. The existence of energy transfer between quinacrine or 33258 Hoechst and 7-amino-actinomycin D in doubly stained chromosomes is demonstrated directly by microfluorometry. The ability of the dyes 7-amino-actinomycin D, methyl green, or netropsin, acting as counterstains, to displace quinacrine, 33258 Hoechst, or chromomycin A3 from chromosomes, is estimated by quantitative analysis of energy transfer data, by photobleaching of the counterstains, or by selective removal of counterstains by appropriate synthetic polynucleotides. Effects on the fluorescence of soluble 33258 Hoechst-DNA complexes due to energy transfer or binding displacement, by actinomycin D or netropsin, respectively, are further differentiated by nanosecond fluorescence decay measurements. Examples are presented of dye combinations for which (a) energy transfer is the primary mechanism operative, (b) binding competition exists, with consequences reinforcing those due to energy transfer, or (c) binding competition is the most important interaction. These analyses of mechanisms responsible for contrast enhancement in doubly stained chromosomes are used to derive information about the relationship between chromosome composition and banding patterns.

Animals↗
PubMed1979

In vivo induction of sister-chromatid exchanges in liver and marrow cells by drugs requiring metabolic activation.

A highly sensitive method for the detection of in vivo induction of sister-chromatid exchange (SCE) has been developed in mice subjected to partial hepatectomy. SCE induction by either acetylaminofluorene (AAF) or cyclophosphamide, drugs requiring metabolic activation, is significantly greater in both regenerating liver and bone-marrow cells of partial hepatectomized animals than in marrow cells of unhepatectomized mice. These experiments have confirmed the ability of AAF, a well known mutagen-carcinogen, to induce SCE formation, even though the cytogenic effects of this drug on non-hepatectomized mice is very small. The in vivo system described has demonstrated the influence of the liver on drug-induced damage to extra-hepatic tissues. The procedures developed should facilitate the detection of drug-induced cytogenic damage and permit the comparison of inter-tissue differences in SCE induction with tissue-specific differences in drug-activation pathways.

2-Acetylaminofluorene↗
PubMed1979

The effect of a tumor promotor, 12-O-tetradecanoyl-phorbol-13-acetate (TPA), on sister-chromatid exchange formation in cultured Chinese hamster cells.

The tumor promoter 12-O-tetradecanoyl-phorbol-13-acetate (TPA) does not increase the sister-chromatid exchange (SCE) frequency in either Chinese hamster ovary (CHO) or lung (V 79) cells which are cultured in the presence of 5-bromodeoxyuridine. Moreover, TPA does not alter the induction of SCEs in CHO cells by mitomycin C during the first 3 cycles following addition of the alkylating agent. These SCE induction data do not by themselves support the hypothesis that tumor promotion by TPA depends on the enhancement of mitotic recombination.

Animals↗
PubMed1979

Late replication in an X-autosome translocation in the mouse: correlation with genetic inactivation and evidence for selective effects during embryogenesis.

A technique involving 5-bromodeoxyuridine, 33258 Hoechst, and fluorescence microscopy has been used to analyze replication kinetics in cells from embryonic and adult mice bearing the Cattanach [T(X;7)ICt] translocation in a balanced or an unbalanced form. In balanced 9- and 13-day female embryos, the translocated X was late replicating in 28 and 22% of the cells, respectively, whereas it was late replicating in only 13% of adult cells. In contrast, in unbalanced females, the translocated X was late replicating in 62 and 70% of 9- and 13-day embryos and in 70% of adult cells. Such divergent late replication frequencies suggest the operation, during development, of selection against cells with extreme genetic imbalance. Within a late-replicating translocated X chromosome, the autosomal segment itself replicated late approximately half of the time, regardless of karyotypic balance. The late replication data are consistent with the measurements of levels of mitochondrial malic enzyme (MOD-2, whose locus is on the autosomal segment) activity in these mice [Eicher E. & Coleman, D. (1977) Genetics 85, 647-658]. The present study also shows a dissociation between the replication timing in X chromatin distal and proximal to the autosomal segment, supporting the hypothesis of at least two inactivation centers in the X chromosome.

Animals↗
PubMed1979

Pairs of fluorescent dyes as probes of DNA and chromosomes.

If two fluorescent dyes with different binding or fluorescence specificities are used simultaneously to stain DNA or chromosomes, the ratio of their fluorescent signals can provide information about base composition or base analogue substitution. Energy transfer between such dye pairs, possible if the fluorescence spectrum of one overlaps the absorption spectrum of the other, can modify observed fluorescence. Microfluorometric measurements were used to document the occurrence of energy transfer between quinacrine or 33258 Hoechst as energy donor and ethidium or 7-aminoactinomycin D as acceptor when used jointly to stain cytologic preparations of human metaphase chromosomes. Use of 7-aminoactinomycin D, a dye with G-C binding specificity, as energy acceptor permitted the identification of human chromosome regions presumptively enriched for clusters of A-T base pairs, based on the resistance of A-T specific fluorescence, from quinacrine or 33258 Hoechst, to energy transfer dependent quenching. The results provide information about basic structural features of metaphase chromosomes, and the associated methodology may prove useful in accentuating specific fluorescent polymorphic chromosome regions.

Cells, Cultured↗