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R J Preston

Publications and source records attributed to R J Preston.

At least 73 records · Page 4Linked to original sources

SCE induction is proportional to substitution in DNA for thymidine by CldU and BrdU.

The incorporation of 5-bromo- and 5-chlorodeoxyuridine into cellular DNA and the induction of sister-chromatid exchanges (SCE) was examined with Chinese hamster ovary cells in culture. Although CldU caused 3-5 times more SCE than BrdU at equal extracellular concentrations, the 2 analogs were incorporated into DNA in place of thymidine similarly over a concentration range of 0.5-20 microM. The results show that SCE induction is linearly proportional to substitution for thymidine by both BrdU and CldU. Finally, based on linear extrapolation, a spontaneous level of approximately 6 SCEs per cell is estimated.

Animals↗

The induction of specific-locus mutations and sister-chromatid exchanges by 5-bromo- and 5-chloro-deoxyuridine.

The induction of 6-thioguanine-resistant (TGr) mutants and sister-chromatid exchanges (SCE) by 5-bromo- and 5-chloro-deoxyuridine (BrdU and CldU) was determined with Chinese hamster ovary cells in culture. Over a concentration range of 0.5-200 microM, CldU treatment caused 3-5 times the number of SCEs found with BrdU; for example, with 10 microM CldU, 60.0 +/- 14.2 SCEs/metaphase (mean +/- S.D.) were induced, while with 10 microM BrdU, only 14.6 +/- 4.1 SCEs/metaphase were found. Despite this difference in SCE induction, both compounds were mutagenic only at concentrations greater than 50 microM, and BrdU was more mutagenic than CldU. These results indicate that SCEs and gene mutations are induced by these thymidine analogs through 2 different mechanisms.

Animals↗

The use of inhibitors of DNA-repair in the study of the mechanisms of induction of chromosome aberrations.

If cells exposed to X-rays or chemical agents are incubated with inhibitors of DNA repair (cytosine arabinoside or aphidicolin), it is possible to accumulate single-strand gaps in the DNA at repairing regions. Upon reversal of the inhibition, these gaps can interact to form chromosome aberrations. It appears that the aberration frequency observed following treatment with radiation or chemical agents is greatly influenced by the rate repair of damage that is converted into aberrations. This argument also extends to considerations of the different cell-cycle-stage sensitivities, the relative sensitivities of different species, and the probability of observing interactive effects between two agents. These aspects are discussed in terms of their contribution to the interpretation of the mechanisms induction of chromosome aberrations.

Aphidicolin↗

X-ray-induced chromosome aberrations in Down lymphocytes: an explanation of their increased sensitivity.

Unstimulated lymphocytes from individuals with Down Syndrome (trisomy 21) are more sensitive to the induction of dicentric and ring aberrations by X rays than normal lymphocytes. Several explanations involving the more rapid rejoining of X-ray--induced lesions in Down cells have been offered. It is shown here that the repair of the DNA damage converted into chromosome aberrations is more rapid in Down cells than normal cells. This more rapid repair results in a higher probability of producing chromosomes aberrations, and hence higher aberration frequencies in Down than normal cells.

Chromosome Aberrations↗

The induction of chromosome-type aberrations in G1 by methyl methanesulfonate and 4-nitroquinoline-N,-oxide, and the non-requirement of an S-phase for their production.

Human lymphocytes were treated in G1 with 4-nitroquinoline-N-oxide (4NQO) and methyl methanesulfonate (MMS) and then incubated in the presence or absence of cytosine arabinoside (ara-C). There was an increase in aberration frequency in those cells incubated with ara-C compared with those treated with 4NQO or MMS alone. This increase was restricted to chromosome-type aberrations. When cells were treated in G2 with 4NQO and then incubated with ara-C until fixation, there was an increase in deletions compared with cells treated with 4NQO alone. No exchange aberrations were observed following any treatment even when deletion frequencies were high, as in the case with 4NQO plus ara-C treatment. These results suggest that ara-C can inhibit the repair of DNA damage induced by 4NQO and MMS that is converted into aberrations. They also show that the terms "S-dependent' and "S-independent' used to describe the modes of action of chemical clastogens are not valid.

4-Nitroquinoline-1-oxide↗

Mammalian in vivo and in vitro cytogenetic assays: a report of the U.S. EPA's gene-tox program.

This report presents an assessment made by the U.S. Environmental Protection Agency Gene-Tox Program's Work Group on mammalian cytogenetics of the clastogenic effects of chemicals in in vivo and in vitro mammalian cell assays. This assessment is based on information provided by the Environmental Mutagen Information Center, Oak Ridge National Laboratory, with the proviso that the experimental protocol used in these papers was adjudged to be acceptable by standards outlined by the Work Group. Some data were accepted as "qualitative only" because the protocol used was fairly close to that proposed as suitable. Using these criteria, 177 papers were selected for review. 6 assays were reviewed: bone marrow (32 papers, 31 chemicals), spermatogonial (10 papers, 10 chemicals), spermatocyte (25 papers, 25 chemicals), oocyte or early embryo (18 papers, 19 chemicals), in vitro cell culture (30 papers, 66 chemicals), and leukocyte (66 papers, 53 chemicals). Each assay was considered separately, and comparisons were then made between them for their similarities or differences in producing a positive or negative clastogenic effect of a particular chemical or chemical class. A large proportion of the available cytogenetic data was not suitable for inclusion in the final data base because of poor experimental design or unsatisfactory reporting of the information. It was not possible to recommend any one assay for determining potential clastogenicity because each had its own particular advantages and limitations and provided unique information. For demonstrating in vivo effects, the bone-marrow assay is probably the simplest and most economical. If only in vitro exposures were considered, leukocytes or cultured mammalian cell lines would be suitable. However, there are advantages to using leukocytes because they are a synchronous population, at least through their cell division, and because of the ready availability of human cells. In general, there was good agreement between clastogenicity and carcinogenicity.

Animals↗

Medium spiny neuron projection from the rat striatum: an intracellular horseradish peroxidase study.

The morphological features of striatal projection neurons and the responses of these neurons to electrical stimulation of the substantia nigra were studied in rats through the methods of intracellular recording and intracellular labeling with the enzyme horseradish peroxidase. Under urethane anesthesia, single striatal neurons were first analyzed for responsiveness to nigral stimuli and then filled electrophoretically with the enzyme in order to permit subsequent serial reconstruction. The axon of the medium spiny neuron was found to form an extensive collateral plexus within the striatum before entering the globus pallidus or the internal capsule. These medium spiny projection neurons responded to nigral stimuli with monosynaptic excitation.

Animals↗

DNA repair and chromosome aberrations: the effect of cytosine arabinoside on the frequency of chromosome aberrations induced by radiation and chemicals.

The frequency of x-ray-induced chromosome aberrations in G0 human lymphocytes was greatly increased when cells were incubated with cytosine arabinoside (ara-C) after irradiation. The frequency of dicentrics increased with increasing ara-C incubation times (one, two, and three hours). Lymphocytes from Down syndrome individuals were more sensitive to aberration induction by x-rays in G0, and the increase in dicentric frequency with ara-C incubation was much more rapid than with normal cells. When G2 normal lymphocytes were x-irradiated and incubated for two or three hours with ara-C until fixation, there was a large increase in deletion frequency compared to cells x-irradiated and incubated in the absence of ara-C. However, no exchanges were observed in the presence of ara-C, compared to 0.29 per cell as when x-rays alone were given. These results form the basis for a discussion of the mechanism of aberration induction by x-rays. Experiments with two chemicals, 4-nitroquinoline-N-oxide and methyl methanesulfonate, show that chromosome-type aberrations can be induced in G1 treated lymphocytes incubated with ara-C. However, these chemicals, in the absence of ara-C incubation, induced no aberrations in G1 at the concentrations used. The mechanism of aberration induction is discussed, particularly in terms of whether or not chemicals can be defined as S-phase dependent.

4-Nitroquinoline-1-oxide↗

The effect of cytosine arabinoside on the frequency of X-ray-induced chromosome aberrations in normal human leukocytes.

The yield of X-ray-induced chromosome aberrations in unstimulated human lymphocytes is greatly enhanced when the cells are incubated with cytosine arabinoside (ara-C) after irradiation. There is an increase in aberration yield with increasing time of incubation in ara-C (1, 2 or 3 h). When G2 cells are X-irradiated and incubated with ara-C until fixation, the deletion yields are considerably increased, but no interchanges are observed. In the absence of ara-C, 0.29 interchanges per cell were observed. The G2 results suggest that ara-C inhibits the repair of damage that leads to the formation of exchange aberrations. The increase in yield in unstimulated lymphocytes is interpreted to be due to an accumulation of strand breaks caused by ara-C inhibition of repair; when the inhibition is reversed with deoxycytidine, these breaks can interact to form aberrations. A possible mechanism for the induction of chromosome aberrations by X-rays is offered on the basis of these results.

Cells, Cultured↗

Presumptive evidence for the absence of functional germ cell chimerism in the marmoset.

Presumptive evidence for functional germ cell chimerism in the marmoset was evaluted by calculating the offspring sex ratio of young derived from animals in which the blood chimerism status was known. In the four possible mating combinations involving chimeric and non-chimeric marmosets, there was no apparent deviation from the anticipated 1:1 sex ratio. Analysis of 2,500 primary spermatocytes from ten known blood chimeras failed to reveal a single cell in which an unequivocal XX bivalent could be identified. The combined breeding and cytogenetic data offer presumptive evidence against functional germ cell chimerism in the marmoset.

Animals↗

Response properties and electrical constants of caudate nucleus neurons in the cat.

1. Response properties and passive electrical constants were assessed for caudate nucleus neurons in pentobarbital-anesthetized cats. The neurons studied were those which could be monosynaptically excited by substantia nigra and thalamic (centromedian-parafascicular) stimulation. 2. Input resistance and membrane time constant were estimated from the plateau value and time course, respectively, of the neuronal membrane response to intracellularly applied current pulses. The average values obtained were 16.5 Momega and 11.3 ms. Specific resistance and capacitance values were calculated. 3. Single or repetitive spikes were readily evoked by nigral or thalamic stimuli or by the application of direct depolarizing currents. Spike thresholds were higher for direct than for synaptic activation (7.2 vs. 5.6 mV). 4. Direct depolarizing stimuli with durations up to 600 ms elicited repetitive discharge with little adaptation of firing rate. The maximum discharge rates induced by direct stimuli were near 200 spikes per second. 5. The intracellular application of tetraethylammonium chloride (TEA) produced spike-prolongation effects in caudate neurons that were similar to the effects reported for other nerve membrane. 6. The probable identity of the recorded neurons as medium spiny neurons was discussed and, in addition, it was proposed that the characteristic silence of these cells is not likely due to intrinsic membrane specialization.

Action Potentials↗

Radiation-induced chromosome aberrations in somatic and germ cells of the male marmoset.

The induction of chromosome aberrations by low LET radiations was studied in peripheral lymphocytes and spermatogonial stem cells of the male marmoset. The data showed that there was not significant difference in the sensitivity of the lymphocytes whether they were irradiated in vitro or in vivo, but the frequency of heritable translocations recovered in the primary spermatocytes were considerably lower than that calculated to occur in the lymphocytes. The data are used to make estimates of human genetic risk from radiation based on limited interspecific comparisons.

Animals↗

The effect of cytosine arabinoside on the frequency of single-strand breaks in DNA of mammalian cells following irradiation or chemical treatment.

1-beta-D-Arabinofuranosyl cytosine (araC), a pyrimidine nucleoside analog used in the treatment of malignant tumors [1, 2], inhibits ultraviolet repair of DNA in a reversible manner. The inhibition occurs during the resynthesis-ligation step and is apparent at all sites undergoing repair. By use of araC it was possible to substantiate the reported observation that the initial velocities of ultraviolet repair are dose dependent and that hamster and human cells are more efficient that mouse cells in excising DNA damage after fluences of less than 50 J/m2. araC does not strongly inhibit gamma-ray-induced repair, although alkali-labile sites are removed more slowly in araC-treated cells. Repair of damage to DNA by N-methyl-N-nitrosoguanidine, mitomycin C, 4-nitroquinoline oxide and 8-hydroxyquinoline is strongly inhibited by araC.

4-Nitroquinoline-1-oxide↗

Electrophysiological and horseradish peroxidase studies of precerebellar afferents to the nucleus interpositus anterior. I. Climbing fiber system.

The input to the nucleus interpositus anterior (NIA) of the cat from the inferior olive (IO) was studied by stimulating the IO while recording intracellularly from the NIA, and by the retrograde transport of horseradish peroxidase (HRP). Stimulation of the IO evoked monosynaptic EPSPs in NIA neurons. The cells labeled in the IO following electrophoretic and pressure injection of HRP into NIA were located in the rostral parts of the dorsal and medial accessory olive. Stimulation of the IO also polysynaptically evoked an IPSP and a late disinhibitory depolarization. Data were presented which indicated that these potentials were mediated by the Purkinje cells of the cerebellar cortex.

Afferent Pathways↗