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L G Littlefield

Publications and source records attributed to L G Littlefield.

At least 37 records · Page 2Linked to original sources
PubMed1993

Induction of micronuclei by bleomycin in G0 human lymphocytes: II. Potentiation by radioprotectors.

Dimethylsulfoxide (DMSO) and WR-1065 are radioprotectors, in that they reduce the effectiveness with which ionizing radiation causes genetic damage. Unlike their protective effects with radiation, these agents potentiate the induction of micronuclei by bleomycin in the cytokinesis-block assay in G0 human lymphocytes. High concentrations of DMSO (1 M) are required to cause potentiation. In contrast, WR-1065 causes dose-dependent potentiation at relatively low concentrations (1.25 to 10 mM). Cytogenetic analysis supports the results from the micronucleus assay, showing higher levels of genetic damage induced by the combination of bleomycin with DMSO or WR-1065 than by bleomycin alone. Possible mechanisms of potentiation are proposed.

Adult↗
PubMed1993

Concentration-dependent protection against X-ray-induced chromosome aberrations in human lymphocytes by the aminothiol WR-1065.

WR-1065, the free-thiol form of WR-2721, has radioprotective effects in various biological systems. We measured the efficiency of WR-1065 in modifying the induction of chromosome aberrations by X rays in human lymphocytes. G0 lymphocytes were incubated for 30 min in medium containing 1-12 mM WR-1065, exposed to 0 or 3.1 Gy 220-kV X rays, washed, and cultured for evaluations of chromosome aberrations and micronuclei (MN). Neither proliferation kinetics nor baseline frequencies of aberrations or MN were affected in nonirradiated cultures incubated in WR-1065 for up to 45 min. Radiation-induced chromosome aberrations and MN varied inversely as a logarithmic function of thiol concentration. At extracellular concentrations of 8-12 mM, WR-1065 protected against > 85% of X-ray-induced chromosome damage as measured by either cytogenetic end point. WR-1065 is more efficient in modulating X-ray-induced chromosome aberrations than dimethyl sulfoxide, which provides protection by scavenging OH radicals. Our data suggest that mechanisms in addition to OH radical scavenging are involved in radioprotection by WR-1065.

Chromosome Aberrations↗
PubMed1991

Frequency of chromosomal aberrations in a subject accidentally exposed to 137Cs in the Goiania (Brazil) radiation accident: intercomparison among four laboratories.

After the Goiania radiation accident which occurred in Brazil in September 1987, an intercomparison was performed to determine whether different cytogenetic laboratories would score similar frequencies of chromosomal aberrations in cultures of lymphocytes of a highly exposed patient. For this purpose 2 chromosome slides from the subject were scored by 4 laboratories in total. The results were consistently close and confirmed the high frequency of chromosome-type aberrations observed initially in the patient.

Accidents, Occupational↗
PubMed1991

Biological effectiveness of neutrons from Hiroshima bomb replica: results of a collaborative cytogenetic study.

The effectiveness of neutrons from a facsimile of the Hiroshima bomb was determined cytogenetically. The "Little-Boy" replica (LBR), assembled at Los Alamos as a controlled nuclear reactor for detailed physical dosimetry, was used. Of special interest, the neutron energy characteristics (including lineal energy) measured 0.74 m from the LBR were remarkably similar to those calculated for the 1945 Hiroshima bomb at 1 to 2 km from the hypocenter, as shown in a companion dosimetric paper (Straume, et al., Radiat. Res. 128, 133-142 (1991)). Thus we examine here the effectiveness of neutrons closely resembling those that the A-bomb survivors received at Hiroshima. Chromosome aberration frequencies were determined in human blood lymphocytes exposed in vitro to graded doses of LBR radiation (97% neutrons, 3% gamma rays). Vials of blood suspended in air at distances up to 2.10 m from the center of the LBR uranium core received doses ranging from 0.02 to 2.92 Gy. The LBR neutrons (E approximately 0.2 MeV) produced 1.18 dicentrics and rings per cell per Gy. They were more effective than the higher-energy fission neutrons (E approximately 1 MeV) commonly used in radiobiology. The maximum RBE (RBEM) of LBR neutrons at low doses is estimated to be 60 to 80 compared to 60Co gamma rays and 22 to 30 compared to 250-kVp X rays. These results provide a quantitative measurement of the biological effectiveness of Hiroshima-like neutrons.

Adult↗
PubMed1990

Chromosome aberrations in relation to radiation dose following partial-body exposures in three populations.

Structural chromosome aberrations were evaluated in peripheral blood samples obtained from three populations exposed to partial-body irradiation. These included 143 persons who received radiotherapy for enlarged thymus glands during infancy and 50 sibling controls; 79 persons irradiated for enlarged tonsils and 81 persons surgically treated for the same condition during childhood; and 77 women frequently exposed as young adults to fluoroscopic chest X rays during lung collapse treatment for tuberculosis (TB) and 66 women of similar ages treated for TB with other therapies. Radiation exposures occurred 30 and more years before blood was drawn. Doses to active bone marrow averaged over the entire body were 21, 6, and 14 cGy for the exposed thymic, tonsil, and TB subjects, respectively. Two hundred metaphases were scored for each subject, and the frequencies of symmetrical (stable) and asymmetrical (unstable) chromosome aberrations were quantified in 97,200 metaphases. Cells with stable aberrations were detected with greater frequency in the irradiated subjects compared with nonirradiated subjects in all three populations, and an overall test for an association between stable aberrations and partial-body ionizing radiation was highly significant (P less than 0.001). We found no evidence that radiation-induced aberrations varied by age at exposure. These data show that exposure of children or young adults to partial-body fractionated radiation can result in detectable increased frequencies of stable chromosome aberrations in circulating lymphocytes 30 years later, and that these aberrations appear to be informative as biological markers of population exposure.

Chromosome Aberrations↗
PubMed1989

Labeling of human centromeres using an alphoid DNA consensus sequence: application to the scoring of chromosome aberrations.

Precise identification of centromeres is required for accurate scoring of asymmetrical chromosome aberrations, such as dicentrics. The centromeric regions of all human chromosomes can be labeled by in situ hybridization of a 30 nucleotide oligomer having the sequence of a conserved region of an alphoid DNA consensus sequence. Fluorescent detection of the hybridized probe allows rapid identification of centromeres and accurate scoring of dicentrics, multicentrics, acentric fragments, and the centromeric content of ring chromosomes. This procedure provides a novel approach for scoring these complex chromosome aberrations, particularly damage induced by radiation or radiomimetic agents.

Centromere↗
PubMed1989

Comparisons of dose-response parameters for radiation-induced acentric fragments and micronuclei observed in cytokinesis-arrested lymphocytes.

We employed cytochalasin B to block cytokinesis in human lymphocytes exposed to 220 kV X-radiation. When 3 micrograms/ml of cytochalasin B was used, most cells escaped the block whereas at 6 micrograms/ml greater than 90% of the mitogen-responsive cells became binucleate. Using the higher concentration of cytochalasin B, we observed a linear-quadratic (i.e. Y = gamma + alpha D + beta D2) dose dependency for X-ray-induced micronuclei (MN) in preparations from three donors. When dose-response parameters were compared with those for total acentrics scored in first division metaphases, we observed no significant differences in estimates of the background (gamma) or linear (alpha) coefficients, but a 2-fold reduction in the beta coefficient for MN. We interpret our data as providing evidence that radiation-induced micronuclei are derived from acentric fragments (AF); that virtually all AF are recovered as MN in binucleate interphase daughter cells at low radiation doses; and that for our data, the relative proportion of AF that will be observed as independent MN decreases by a constant factor of approximately one-half as radiation dose increases.

Adult↗
PubMed1989

Telomere staining of human chromosomes and the mechanism of radiation-induced dicentric formation.

The majority of models of radiation action developed over the past half century hold that the curvilinear dose responses exhibited by eukaryotic cells to sparsely ionizing radiations result from the interaction of pairs of lesions produced in sensitive targets of the cell. Within this conceptual framework, chromosomal exchange aberrations (e.g., interchanges) are believed to occur through the interaction of damaged sites on both chromosomes participating in the exchange. In contrast, the model proposed by Chadwick and Leenhouts (as well as some other models) suggests that such exchanges arise from initial radiation damage to only one chromosome, which then becomes associated with an undamaged chromosome. A particular aspect of this theory is that asymmetrical exchanges, such as dicentrics, may be formed from the rejoining of a broken end of one chromosome to the telomere of another. By using a DNA probe that specifically hybridizes to the telomeric region of human chromosomes, we were able to test this assertion directly. After scanning more than 200 dicentrics produced in normal human fibroblasts by 6 Gy of 60Co gamma rays, virtually none were found that contained telomeres located between the centromeres of this aberration type. Therefore, since the proposed telomere-break rejoining process, per se, is not necessarily a central element of the Chadwick-Leenhouts model, we suggest the theory be modified to exclude this mechanism.

Chromosome Aberrations↗
PubMed1989

Chromosome aberrations in peripheral lymphocytes and radiation dose to active bone marrow in patients treated for cancer of the cervix.

An international study of cervical cancer patients reported a doubling of the risk for leukemia following radiotherapy. To evaluate the extent of residual chromosome damage in circulating T-cell lymphocytes in this population, approximately 200 metaphases were examined from each of 96 irradiated and 26 nonirradiated cervical cancer patients treated more than 17 years ago (average 23 years). Radiation dose averaged over the total red bone marrow was estimated to be 8.1 Gy. The type and frequency of stable and unstable chromosome aberrations were quantified in 24,117 metaphases. Unstable aberrations did not differ significantly between irradiated and nonirradiated patients (P greater than 0.5). Stable aberrations (i.e., translocations, inversions, or chromosomes with deleted segments), however, were significantly higher among irradiated (2.8 per 100 cells) compared to nonirradiated (0.7 per 100 cells) women (P less than 10(4). The frequency of these stable aberrations was found to increase significantly with increasing dose to the bone marrow. These data indicate that a direct relationship between radiation dose and extent of damage to somatic cells persists in populations and can be detected many years after partial-body radiation exposure. The stable aberration rate in irradiated cervical cancer patients was 50 to 75% lower than those observed 25 years or more after radiation exposure in atomic bomb survivors and in ankylosing spondylitis patients treated with radiotherapy. The average marrow dose was only 1 Gy in the examined atomic bomb survivors and 3.5 Gy in the ankylosing spondylitis patients. It appears, then, that a very high dose delivered to the pelvic cavity in fractionated doses resulted in far fewer persistent stable aberrations than lower doses delivered either in acute whole-body exposure or in fractionated doses to the spinal column and sacroiliac joints. The higher radiation dose and the concentration of that dose in a smaller area of the body appear to be responsible for the lower rate of persistent aberrations observed in cervical cancer patients.

Bone Marrow↗
PubMed1988

Current status of cytogenetic procedures to detect and quantify previous exposures to radiation.

The estimation of the magnitude of a dose of ionizing radiation to which an individual has been exposed (or of the plausibility of an alleged exposure) from chromosomal aberration frequencies determined in peripheral blood lymphocyte cultures is a well-established methodology, having first been employed over 25 years ago. The cytogenetics working group has reviewed the accumulated data and the possible applicability of the technique to the determination of radiation doses to which American veterans might have been exposed as participants in nuclear weapons tests in the continental U.S.A. or the Pacific Atolls during the late 1940s and 1950s or as members of the Occupation Forces entering Hiroshima or Nagasaki shortly after the nuclear detonations there. The working group believes that with prompt peripheral blood sampling, external doses to individuals of the order of about 10 rad (or less if the exposure was to high-LET radiation) can accurately be detected and measured. It also believes that exposures of populations to doses of the order of maximum permissible occupational exposures can also be detected (but only in populations; not in an individual). Large exposures of populations can also be detected even several decades after their exposure, but only in the case of populations, and of large doses (of the order of 100 to several hundred rad). The working group does not believe that cytogenetic measurements can detect internal doses from fallout radionuclides in individuals unless these are very large. The working group has approached the problem of detection of small doses (less than or equal to 10 or so rad) sampled decades after the exposure of individuals by using a Bayesian statistical approach. Only a preliminary evaluation of this approach was possible, but it is clear that it could provide a formal statement of the likelihood that any given observation of a particular number of chromosomal aberrations in a sample of any particular number of lymphocytes actually indicates an exposure to any given dose of radiation. It is also clear that aberration frequencies (and consequently doses) would have to be quite high before much confidence could be given to either exposure or dose estimation by this method, given the approximately 3 decades of elapsed time between the exposures and any future blood sampling.(ABSTRACT TRUNCATED AT 400 WORDS)

Abnormalities, Radiation-Induced↗
PubMed1988

Modulation of radiation-induced chromosome aberrations by DMSO, an OH radical scavenger. 1: Dose-response studies in human lymphocytes exposed to 220 kV X-rays.

Human G0 lymphocytes were exposed to 220 kV X-radiation in the presence or absence of DMSO, an efficient selective scavenger of OH radicals. Our studies demonstrate that DMSO affects a concentration-dependent modulation of induced asymmetrical aberrations in human lymphocytes exposed to approximately 3.0 Gy, with maximum protectible fractions of approximately 70 percent at DMSO concentrations of greater than or equal to 1 M. The dose dependency for dicentrics in lymphocytes acutely exposed to X-ray doses of 0.51 to 4.98 Gy in the absence of DMSO is adequately described by the linear-quadratic dose-response function Y = alpha D + beta D2. Data from duplicate cultures exposed in the presence of 1 M DMSO produce an excellent fit to the regression function modified as follows: Y(+ DMSO) = alpha(delta D) + beta(delta D)2 where the 'dose modifying' factor delta = 0.501. We interpret these findings as providing evidence that OH radical-mediated lesions in DNA account for approximately 50 percent of the dose dependency for dicentrics resulting from either one-track or two-track events, following exposures of non-cycling cells to moderate-to-high doses of low LET radiation. These data may be used in additional calculations to derive an estimate of approximately 6 x 10(8) s-1 for the rate of reaction of OH radicals with DNA targets involved in aberration formation.

Chromosome Aberrations↗
PubMed1987

Cytogenetic evaluations in human lymphocytes exposed to methyl acetimidate, a lysine-specific protein crosslinking agent.

The cytogenetic effects of methyl acetimidate (MAI), a lysine-specific protein crosslinking reagent, were investigated using human peripheral lymphocytes in culture. Lymphocytes were treated with the chemical either prior to PHA exposure or 2-3 days following mitogenic stimulation and assessed for perturbations in cellular proliferation and induction of SCEs. Severe reductions in the mitotic index (MI) and pronounced decreases in the proportion of metaphases proceeding beyond M(1) were observed following G0 exposure to MAI concentrations of as low as 2 mM; with complete suppression of mitotic activity in all cultures exposed to levels of 3 mM MAI or greater. Concentrations resulting in severe depression in MI caused only moderate increases in SCEs. Cells exposed to less than 10 mM MAI during the late S-G2 stages of the cell cycle and harvested at the first metaphase following treatment exhibited profound mitotic delay, impaired prophase to metaphase transitions and abnormal mitotic configurations. These findings demonstrate that protein-specific crosslinking agents may induce a wide spectrum of adverse cytogenetic outcomes in both cycling and noncycling lymphocytes.

Cell Cycle↗
PubMed1986

Analysis of chromosome aberrations in lymphocytes of long-term heavy smokers.

We assessed the incidence of structural chromosome aberrations in 500 diploid first-division metaphases from 48-h lymphocyte cultures from each of 6 non-smokers and from 6 persons who had smoked a minimum of 1 pack of cigarettes per day for at least 20 years. Cytogenetic analyses of coded slides revealed a single dicentric chromosome with its accompanying fragment and two symmetrical chromatid exchanges in 3000 metaphases from the non-smokers. In contrast, 9 dicentric chromosomes, 8 translocations or inversions, and 7 chromatid exchanges were observed in 3000 metaphases from lymphocyte cultures from the 6 heavy smokers. A total of 13 metaphases having chromosome-type inter- or intra-changes was noted including 9 with a single aberration, and 4 with 2 or more. Our findings provide additional evidence of the in vivo clastogenicity of cigarette smoke in long-term heavy smokers, and further demonstrate that the distribution of chromosome-type exchange aberrations is overdispersed relative to that expected based on Poisson assumptions.

Adult↗
PubMed1985

Premature separation of centromeres in marrow chromosomes from an untreated patient with acute myelogenous leukemia.

Cytogenetic studies in an untreated patient with acute myelogenous leukemia revealed that greater than 60% of his marrow metaphases had pronounced anomalies in the centromere region. In the least affected chromosomes, the centromere appeared as a nonstaining hole, whereas in the most affected chromosomes, it appeared that the centromere segment had prematurely migrated at right angles to the telomeric portions. The population of mitoses exhibiting premature separation of the centromeres was reduced to virtually none during periods of drug induced remission. Our findings suggest that cytogenetic changes in malignant myeloid cells may be characterized by defects in centromere separation, as well as by alternations in karyotype.

Bone Marrow↗
PubMed1984

Persistence of SCE-inducing lesions after G0 exposure of human lymphocytes to differing classes of DNA-damaging chemicals.

We conducted studies to determine whether cycling human lymphocytes are equally efficient in repairing sister chromatid exchange (SCE)-producing lesions induced by differing classes of DNA-damaging chemicals. Lymphocytes were pulse-treated during G0 with mitomycin C (MMC), N,N',N''-triethylenethiophosphoramide (ThioTEPA), ethylmethanesulfonate (EMS), or cis-diamminedichloroplatinum (cis-DDP). Bromodeoxyuridine (BrdUrd) was added to the 72 hr cultures at 0 hr or at 48 hr after phytohemmagglutinin stimulation. The concentrations of chemicals employed induced a greater than 2-fold increase in SCEs in second-division metaphases from lymphocytes cultured in the presence of BrdUrd for the entire 72 hr. The analysis of SCEs in uniformly harlequinized metaphases from G0-treated lymphocytes cultured in BrdUrd for the terminal 24 hr showed no increase above baseline after exposure to MMC, and intermediate increases above baseline after exposures to ThioTEPA and cis-DDP. However, after G0 treatment with EMS, the observed SCE frequency was consistent with that expected had all DNA lesions persisted and continued to give rise to SCEs during 3 cell cycles. These findings suggest that cycling human lymphocytes are not equally efficient in eliminating SCE-producing lesions after exposure to differing classes of DNA-damaging chemicals.

Alkylating Agents↗
PubMed1984

Evaluation of chemically induced cytogenetic lesions in rabbit oocytes. III. A postimplantation analysis of streptonigrin effects.

The frequency of consistent structural chromosome aberrations was determined for 108 fetuses (day 14 of gestation) from 20 female rabbits treated before conception with 90 micrograms/kg of streptonigrin (NSC-45383). The findings by karyotype analysis of 1.85% of the fetuses with consistent aberrations was compared with 6.32% frequency previously observed in 6-day (preimplantation) blastocysts from identically treated females and found to be significantly reduced. The interpretation of the findings is that the developmental events of implantation and placentation are effective in eliminating from further gestation the majority of the rabbit conceptuses with consistent structural chromosome abnormalities. The data are also discussed relative to the predictive nature of somatic cell chromosome damage in women of childbearing age being an estimator of risk of producing offspring with structural chromosome aberrations. The data show that preconceptional treatment of female rabbits with streptonigrin caused a shift in the cytogenetically determined sex ratio of the 14-day fetuses, but not of the 6-day blastocysts. The significant decrease in relative numbers of males observed is consistent with the induction of lethal mutations on the X chromosome. This finding is discussed in conjunction with the timing and parental specificity of X chromosome inactivation during mammalian embryonic development.

Animals↗