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The recently published NCRP Commentary No. 27 evaluated the new information from epidemiologic studies as to their degree of support for applying the linear nonthreshold (LNT) model of carcinogenic effects for radiation protection purposes (NCRP 2018 Implications of Recent Epidemiologic Studies for the Linear Nonthreshold Model and Radiation Protection, Commentary No. 27 (Bethesda, MD: National Council on Radiation Protection and Measurements)). The aim was to determine whether recent epidemiologic studies of low-LET radiation, particularly those at low doses and/or low dose rates (LD/LDR), broadly support the LNT model of carcinogenic risk or, on the contrary, demonstrate sufficient evidence that the LNT model is inappropriate for the purposes of radiation protection. An updated review was needed because a considerable number of reports of radiation epidemiologic studies based on new or updated data have been published since other major reviews were conducted by national and international scientific committees. The Commentary provides a critical review of the LD/LDR studies that are most directly applicable to current occupational, environmental and medical radiation exposure circumstances. This Memorandum summarises several of the more important LD/LDR studies that incorporate radiation dose responses for solid cancer and leukemia that were reviewed in Commentary No. 27. In addition, an overview is provided of radiation studies of breast and thyroid cancers, and cancer after childhood exposures. Non-cancers are briefly touched upon such as ischemic heart disease, cataracts, and heritable genetic effects. To assess the applicability and utility of the LNT model for radiation protection, the Commentary evaluated 29 epidemiologic studies or groups of studies, primarily of total solid cancer, in terms of strengths and weaknesses in their epidemiologic methods, dosimetry approaches, and statistical modelling, and the degree to which they supported a LNT model for continued use in radiation protection. Recommendations for how to make epidemiologic radiation studies more informative are outlined. The NCRP Committee recognises that the risks from LD/LDR exposures are small and uncertain. The Committee judged that the available epidemiologic data were broadly supportive of the LNT model and that at this time no alternative dose-response relationship appears more pragmatic or prudent for radiation protection purposes.

The Rudh’ a Chromain sill is a composite minor intrusion emplaced into Jurassic sandstones on the south coast of the Ross of Mull, NW Scotland, and is part of a suite of basic–silicic sheets associated with the early development of the 58–56 Ma Mull Central Igneous Complex. New whole-rock major and trace element data combined with Sr–Nd–Pb isotope data indicate that two distinct magma end members were involved in the formation of the sill: a tholeiitic basaltic andesite magma generated by contamination and fractional crystallization of regionally available olivine tholeiite basaltic magma, and a rhyolitic magma produced predominantly through crustal melting of basement metasediments. Intermediate compositions at the gradational boundaries between the basic margins and silicic interior of the sill formed by high temperature diffusive hybridization within a compositionally-zoned magma reservoir prior to sheet emplacement. The basic portions of the sheet are replete with a large variety of crustal xenoliths, as well as numerous gabbroic and noritic cumulate fragments. The Rudh’ a’ Chromain sill therefore preserves evidence for the complex interplay between a number of magmatic processes which, when combined with data from the remainder of the suite, suggests that the magmatic plumbing system which fed the sill complex was not simply one large, long-lived magma chamber, but rather a plexus of variably connected sheet-like magma reservoirs.

The Cnoc Rhaonastil minor dolerite intrusion on Islay, NW Scotland represents a single body of alkali olivine basalt magma which differentiated in situ, from leucodolerite, through teschenite to minor nepheline syenite. The syenites occur as isolated nests and pegmatitic schlieren within the leucodolerite, and schlieren of gabbroic pegmatite also occur at the margin of the teschenite. The differentiated rocks contain pyroxene, amphibole and biotite of variable compositions which reflect both primary fractionation processes and late-stage deuteric alteration and reaction.Mafic phases within the gabbroic pegmatite, teschenite and syenite are typically rimmed and speckled with biotite, the composition of which is controlled by the local environment of crystallization. The nepheline syenites contain primary ferro-kaersutite which, where in contact with interstitial patches, has been altered to arfvedsonite, which occasionally contains up to 1.2 wt.% ZrO2. The occurrence of Zr-arfvedsonite (and of Zr-aegirine) in interstitial patches suggests that variably trace element-enriched domains existed within the residual melts on very small scales.

This study examined the interactive effects of pregnancy and aerobic conditioning on maternal cardiac structure and function. Effects of closely monitored cycle ergometer conditioning were studied during the second (TM2) and third trimesters (TM3) in 22 previously sedentary pregnant women (exercised group, EG) and a nonexercising pregnant control group with similar characteristics (CG, n = 19). Subjects were studied in the resting state by two-dimensional echocardiography and during cycle ergometer exercise at three steady-state power outputs at the start of TM2 (ENTRY), at the end of TM2 and TM3 (postconditioning), and 3-4 months postpartum (NPR, nonpregnant reference, CG only). Aerobic conditioning did not increase left ventricular dimensions beyond those attributable to pregnancy itself. In addition, in contrast with previous studies of nonpregnant women, physical conditioning during pregnancy did not reduce heart rate (HR) in the resting state. During exercise, the slope of the HR versus oxygen uptake (Vo2) regression decreased significantly between preconditioning and the end of TM3 in the EG, suggesting that training-induced reductions in HR become more evident with increasing exercise intensity. Also, significant reductions in oxygen pulse (Vo2/HR) were observed at all three work rates in the CG, but not in the EG. These findings support the hypothesis that the cardiovascular effects of aerobic conditioning are obscured by more powerful effects of pregnancy in the resting state but become "unmasked" during strenuous exercise.Key words: human gestation, cycle ergometer exercise, echocardiography, heart rate, stroke volume.

Plagioclase-rich reaction zones occur around numerous aluminous crustal xenoliths within a suite of Palaeogene sub-volcanic basic sheets on the Isle of Mull, NW Scotland. The xenoliths consist of a glassy core, containing mullite needles, generated from the melting of pelitic source rocks. Thick plagioclase mantles grew at the interface between the aluminous liquid and the enclosing basaltic magma and provide a high-level analogue for the petrogenesis of Proterozoic massif-type anorthosites. Similar interactions between mantle-derived basic magmas ponded at the base of the crust and relatively Al-rich lower crustal lithologies would result in the precipitation of large volumes of plagioclase. Anorthosite massifs were then emplaced at higher crustal levels as crystal-rich mushes within relatively juvenile Proterozoic crust. The model negates the need to crystallize large volumes of mafic minerals prior to the production of plagioclase-saturated liquids, and also accounts for the significant influence of crustal sources on the isotopic signatures of all members of the anorthosite suite.

The Cnoc Rhaonastil dolerite boss on Islay, NW Scotland represents a body of alkali-olivine basalt magma which differentiated at low pressure and in situ, from dolerite through teschenite to minor nepheline-syenite. The syenites occur as isolated pods and pegmatitic schlieren within the leucodolerite, and have an exotic mineralogy including Zr-aegirine, Zr-arfvedsonite, Ca-catapleiite, zirconolite and aenigmatite. Fluor-apatite occurs as an accessory phase in the dolerite, but becomes more abundant within the teschenite and syenites. Total REE contents within apatites in the dolerites are typically low (σREE = 0·57–3·21 wt.% oxide), the highest REE contents occurring in irregular, deuterically altered rims and internal patches. The REE-enriched rims also have slightly elevated SiO2 contents at 0·81–0·95 wt.%, suggesting that the substitution scheme Ca2++P5+ ⇔ REE3++Si4+ was operating. These apatites have up to 0·08 wt.% Cl and 3·7 wt.% F, with most being almost pure end-member fluor-apatite. The majority of the teschenite apatites show the least REE-enrichment (σREE = 0·27–0·45 wt.%), coupled with low Na (2 (2O, implying that the substitution scheme Na+ + REE3+⇔2Ca2+ dominated over the more typical scheme involving Si4+ which operated in the dolerites and teschenite. Other zones are either variably enriched in Y (up 2·1 wt.% Y2O3) or Th (up to 0·85 wt.% ThO2). However, there is no correlation between Y and REE contents, suggesting that crystallographic factors were involved in apatite Y and REE partitioning. The REE-rich apatites have very low Cl content (

Non-genotoxic chemical carcinogens are capable of inducing tumors in rodents without interacting with or directly altering the genetic material. Since a preponderance of evidence suggests that cancer results from the accumulation of genetic alterations, the mechanisms by which many non-genotoxic carcinogens induce genotoxic events remain unclear. The present study investigated whether the mitogenic, non-genotoxic carcinogen phenobarbital (PB) could alter cell-cycle checkpoint controls, thereby indirectly leading to the accumulation of genetic damage. Initial studies involved characterizing cell-cycle checkpoint responses to DNA damage in freshly isolated B6C3F1 mouse hepatocytes. These cells responded to bleomycin-induced DNA damage by arresting in G1 and G 2 . Cell-cycle arrest was coupled with p53 protein induction ; however, p21 WAFI protein levels remained unchanged. Studies that utilized hepatocytes isolated from C57BL p53 -/- mice showed that the DNA damage-induced G 1 cell-cycle arrest was dependent on p53 function, but cell-cycle arrest in G 2 was not affected by loss of p53. PB was able to delay and attenuate the G 1 checkpoint response without altering G 2 checkpoint function. A reduction in p53 protein, but not transcript levels, was observed in hepatocytes exposed to PB. Additionally, PB delayed and attenuated p53 protein induction during DNA damage, which suggests that changes in the p53 protein may be contributing to the attenuated G 1 checkpoint response caused by PB. Altered G 1 checkpoint function represents an epigenetic mechanism by which phenobarbital may prevent the detection and repair of DNA damage and indirectly increase the frequency of genotoxic events above that occurring spontaneously. Abrogation of checkpoint controls may, thus, play an important mechanistic role in mitogenic, non-genotoxic chemical carcinogenesis.

Intruded into the Palaeogene lava field and underlying Moine (Neoproterozoic) crystalline basement rocks around Loch Scridain, Isle of Mull, Scotland, is a suite of high-level, inclined, xenolithic sheets, ranging in composition from basalt, through andesite and dacite, to rhyolite. These sheets, associated with the Mull central volcano, were emplaced post 55 Ma. As well as numerous crustal xenoliths, the more basic members of the complex contain a diverse suite of ultrabasic and basic xenoliths. Xenolith types include feldspathic peridotite with cumulus olivine, pyroxenite, gabbro with cumulus plagioclase and cumulus clinopyroxene, and pure anorthosite. Mineralogical data, coupled with whole-rock major- and trace-element data from a small number of the xenoliths suggest that the xenoliths represent early-formed cumulates cognate with their host basalts. Sr and Nd isotope data from the xenoliths confirms the cognate origin, and also shows that the basic magmas suffered crustal contamination at an early stage.

In this report, the Commission recommends approaches to national authorities for their definition of the scope of radiological protection control measures through regulations, by using its principles of justification and optimisation. The report provides advice for deciding the radiation exposure situations that should be covered by the relevant regulations because their regulatory control can be justified, and, conversely, those that may be considered for exclusion from the regulations because their regulatory control is deemed to be unamenable and unjustified. It also provides advice on the situations resulting from regulated circumstances but which may be considered by regulators for exemption from complying with specific requirements because the application of these requirements is unwarranted and exemption is the optimum option. Thus, the report describes exclusion criteria for defining the scope of radiological protection regulations, exemption criteria for planned exposure situations, and the application of these concepts in emergency exposure situations and in existing exposure situations. The report also addresses specific exposure situations such as exposure to low-energy or low-intensity adventitious radiation, cosmic radiation, naturally occurring radioactive materials, radon, commodities, and low-level radioactive waste. The quantitative criteria in the report are intended only as generic suggestions to regulators for defining the regulatory scope, in the understanding that the definitive boundaries for establishing the situations that can be or need to be regulated will depend on national approaches.

Chronic hypoxia caused by migration of native sea-level dwellers to high altitude or chronic lung disease leads to the development of increased pulmonary vascular resistance and pulmonary hypertension. This altitude-induced hypertension offers no obvious benefit and may indeed be maladaptive. A major mechanism thought to contribute to the development of pulmonary hypertension is hypoxia-induced loss of small blood vessels, sometimes termed rarefaction or pruning. More recent evidence caused us to question this widely accepted concept including the potent angiogenic effect of chronic hypoxia in all other vascular beds and the demonstration that new vessels can form in the pulmonary circulation when stimulated by chronic infection and lung resection. We tested the hypothesis that chronic environmental hypoxia causes angiogenesis in the adult pulmonary circulation by using stereological techniques combined with confocal microscopy to examine the resultant changes in pulmonary vascular structure in rats. We found that chronic hypoxia resulted in increased total pulmonary vessel length, volume, endothelial surface area and number of endothelial cells in vivo. This is the first reported demonstration of hypoxia-induced angiogenesis in the mature pulmonary circulation, a structural adaptation that may have important beneficial consequences for gas exchange. These findings imply that we must revise the widely accepted paradigm that hypoxia-induced loss of small vessels is a key structural change contributing to the development of pulmonary hypertension in high altitude adaptation and chronic lung disease.

This study examined the interactive effects of pregnancy and aerobic conditioning on maternal cardiac structure and function. Effects of closely monitored cycle ergometer conditioning were studied during the second (TM2) and third trimesters (TM3) in 22 previously sedentary pregnant women (exercised group, EG) and a nonexercising pregnant control group with similar characteristics (CG, n = 19). Subjects were studied in the resting state by two-dimensional echocardiography and during cycle ergometer exercise at three steady-state power outputs at the start of TM2 (ENTRY), at the end of TM2 and TM3 (postconditioning), and 3-4 months postpartum (NPR, nonpregnant reference, CG only). Aerobic conditioning did not increase left ventricular dimensions beyond those attributable to pregnancy itself. In addition, in contrast with previous studies of nonpregnant women, physical conditioning during pregnancy did not reduce heart rate (HR) in the resting state. During exercise, the slope of the HR versus oxygen uptake (VO2) regression decreased significantly between preconditioning and the end of TM3 in the EG, suggesting that training-induced reductions in HR become more evident with increasing exercise intensity. Also, significant reductions in oxygen pulse (VO2/HR) were observed at all three work rates in the CG, but not in the EG. These findings support the hypothesis that the cardiovascular effects of aerobic conditioning are obscured by more powerful effects of pregnancy in the resting state but become "unmasked" during strenuous exercise.

Cytogenetic assays are an integral component of the battery of short-term assays that are used for the hazard identification component of a cancer risk assessment. The protocol for the conduct of such assays for maximal sensitivity for detecting clastogenicity has to be attendant to the mechanism of induction of the endpoint being assessed and the fact that several aberration types are cell lethal necessitates that analysis be for cells at their first posttreatment metaphase. Cytogenetic assays for human populating monitoring have been used for predicting potential for carcinogenicity in humans. However, the assays as typically conducted are not appropriate for chronic exposures because nontransmissible alterations are assessed. The use of fluorescent in situ hybridization (FISH) techniques for the assessment of transmissible changes such as reciprocal translocations are required to make population monitoring studies interpretable, and for removing some of the concern over the influence of confounders on outcome. The database for the cytogenetic effects of ethylene oxide in vitro and in vivo, with an emphasis on human population monitoring, has been critically reviewed. Based on the endpoints studied, the size of the study groups, the information on exposure, the nature of any exposure response data, and the possible influence of confounders (i.e., control matching), it is concluded that acute, high exposures to ethylene oxide with sampling shortly (a few days) after exposure can be detected by increases in chromosome aberrations or SCE in peripheral lymphocytes. Such increases are indicators of exposure to a genotoxic chemical and not predictors of subsequent adverse health effects to individuals. The effect of chronic and/or low level (less than about 25 ppm) exposures cannot be reliably evaluated using current methods. The use of FISH, for example, for assessing reciprocal translocation frequencies (as a measure of transmissible events) will greatly improve the ability to detect chronic exposures to clastogenic chemicals.

The purposes of this review were twofold: to apply modern physicochemical principles to explain changes in acid-base regulation and the control of ventilation in human pregnancy; and to demonstrate the value of pregnancy as a model for the study of endocrine effects on physiological control systems. Application of P.A. Stewart's approach (P.A. Stewart. Can. J. Physiol. Pharmacol. 61: 1444-1461, 1983) shows that lower values of plasma hydrogen ion concentration ([H+]) observed at rest and in association with exercise in pregnancy are the result of lower values for carbon dioxide tension (Pco2) and total weak acid ([A(tot)]). This effect is partly offset by a lower strong ion difference ([SID]). The ability to predict plasma [H+] at rest and following strenuous exercise in pregnancy (J.G. Kemp, F.A. Greer, and L.A. Wolfe. J. Appl. Physiol. 83: 644-651, 1997) supports the validity of Stewart's approach. Jennings and associates (D.B. Jennings. Can. J. Physiol. Pharmacol. 72: 1499-1512, 1994) have further demonstrated in animal models the involvement of plasma osmolality and circulating levels of angiotensin II (ANG II) and arginine vasopressin (AVP) in the chemical control of ventilation. We hypothesize that pregnancy-induced increases in respiratory sensitivity to carbon dioxide are the combined result of reduced plasma osmolality, reduced cerebrospinal fluid [SID], and augmented circulating levels of progesterone, ANG II, and AVP.

Telomeres in most species consist of repeat units of a small number of nucleotides that together with secondary structures and associated proteins stabilize the linear chromosomal DNA molecule. Chromosomes lose a small amount of telomeric DNA after each cell replication. It has been proposed that when telomeres shorten below a critical length, a DNA damage response pathway is activated and induces cell cycle arrest. In cells such as stem cells that maintain a proliferative capacity, telomere length is maintained by the reverse transcriptase, telomerase. In addition, telomerase activity is present in 90% of primary human tumors, suggesting a role for telomerase in providing a proliferative capacity to cells, which is a requirement in progression to malignancy. Telomerase activity can be involved in chromosome healing, although telomerase-independent processes also appear to be capable of capping broken chromosome ends. This review describes the structure and maintenance of telomeres, the importance of a critical telomere length to cell proliferation and the telomeric status of broken chromosome ends produced during development or by spontaneous or induced DNA damages.

This paper describes the four cytogenetic endpoints most frequently used in hazard identification assays as the first step in the risk assessment process. These are structural chromosome aberrations, micronuclei, aneuploidy, and sister chromatid exchanges. The biological mechanisms involved in the formation of the alterations observed in each assay are briefly discussed. Variations in and recent improvements to each assay are described, with an emphasis on the use of molecular techniques to improve the sensitivity of the assay, and to allow for detection of specific alterations that are, or could be, associated with cancer induction. This, in turn, will make the data obtained in the cytogenetic assays more useful in cancer and genetic risk assessment. Thus, the aim of this paper is to encourage cytogeneticists to design their experiments in such a way that the data obtained will be of maximum possible benefit for characterizing and quantifying adverse human health effects, particularly cancer.

Results from new chromosome studies in laboratory animals, comparative investigations of styrene metabolism and pharmacokinetics in humans and animals, and several recent cytogenetic surveys of styrene-exposed workers have necessitated a comprehensive re-evaluation of the chromosome-damaging effects of this chemical. Both styrene and its genotoxic metabolite, styrene oxide, can induce chromosome aberrations (CA) and sister chromatid exchanges (SCE) in vitro, but the chromosome-damaging ability of styrene is only manifested if test conditions favour its metabolic activation over inactivation. There is no convincing evidence of styrene clastogenicity in experimental animals. Styrene oxide is clastogenic only at lethal concentrations via i.p. injection in Chinese hamsters (but not via inhalation) or after oral treatment of mice, a route considered inappropriate for investigating the chromosome-damaging potential of inhaled styrene in man. Styrene and styrene oxide can induce SCE in animals at very high concentrations. Eighteen of 52 cytogenetic studies (CA, micronuclei, SCE) on peripheral blood lymphocytes of styrene workers have reported increases in chromosome damage. The positive findings are not compatible with the conclusion that styrene is responsible for the cytogenetic effects for the following reasons. 1. (a) The positive or negative outcome of the various investigations bears no relationship to the degree of exposure of the workers. 2. (b) There is no convincing evidence of a positive dose response relationship. 3. (c) The relative induction of CA and SCE in worker studies are the opposite of observations of styrene effects in cultured lymphocytes and in laboratory animals. 4. (d) The reports of chromosome-type exchanges in some studies of styrene workers is inconsistent with observations of styrene clastogenicity in cultured lymphocytes. 5. (e) Reports of SCE induction in workers exposed to low concentrations of styrene are not compatible with results of animal inhalation studies, particularly in view of the differences in styrene metabolism and pharmacokinetics between humans and rodents. The increases in cytogenetic effects reported in some studies on styrene workers are probably attributable to the presence of other chromosome-damaging agents in the workplace and/or to inadequate investigations.

A large number of studies have been reported on the genotoxicity of styrene in vitro and in vivo and the potential effects on humans of occupational exposure. Because of a variety of technical problems and difficulties in data interpretation, it has not been clearly established whether styrene can induce chromosomal aberrations and/or sister chromatid exchange (SCE) in vivo in animals or humans. The importance of clarifying this situation led to the development of the study described in this paper. Male Fischer 344 rats were exposed to styrene at concentrations of 150, 500 or 1000 ppm for 6 h/day on 5 days/week for 4 weeks. A negative control (air) was included. An additional control (ethylene oxide, 150 ppm) group was included in an attempt to establish the usefulness of rat lymphocytes for cytogenetic analysis in this protocol of long-term exposure by inhalation. The choice of agent and of exposure was based on the expectation that they would produce a positive response for SCE and/or chromosomal aberrations under the assay conditions used. Peripheral blood samples were drawn at 1, 2, 3 and 4 weeks of exposure and at 4 weeks after the end of exposure. Cultures were established, and SCE (second mitosis) and chromosomal aberrations (first mitosis) were analysed. The frequency of chromosomal aberrations was not increased over that in the air controls in the animals exposed to styrene or ethylene oxide at any of the sampling times. Styrene did not induce SCE at any of the concentrations or sampling times; however, the frequency of SCE was increased following exposure to ethylene oxide at all sampling times, with a positive exposure-response relationship with time of exposure as the variable. The data are compared with other, similar sets reported in the literature, and their significance for predicting responses in people occupationally exposed to styrene is discussed.

In order to study the mechanism of induction of mutations and chromosome aberrations by ionizing radiations, it is particularly useful to have available radiation-sensitive mutants. While several X-ray-sensitive rodent cell lines are available, they have been selected rather nonspecifically. It was determined that selection for resistance to the DNA replication/repair inhibitor, 1-beta-D-arabinofuranosylcytosine (ara-C), would permit production of a set of X-ray-sensitive mutant cell lines that would be defective in the resynthesis step of excision or recombination repair. Such mutant cells could also be used for the isolation and characterization of human DNA repair genes. In particular, it was predicted that the repair gene defective in individuals with ataxia telangiectasia (AT) might be amenable to study with ara-C-resistant (X-ray-sensitive) mutants, since additional studies, presented here, have shown that AT cells are resistant to ara-C. In the long term, it is hoped that determining the specific defect in AT might lead to an understanding of the possible role of defective repair in tumor induction and/or progression. The general approach used to isolate ara-C-resistant Chinese hamster ovary cell mutants was to treat cells with ethyl methanesulfonate and select in increasing concentrations of ara-C. Although several mutants were isolated, one in particular, Ara-CR213, has been studied most extensively. It was selected largely because it shows the greatest sensitivity to X-rays. Ara-CR213 cells were hypersensitive to the killing effect of X-rays with an LD10 of 2.5 Gy as compared to the wild-type cells that had an LD10 of 6 Gy. The mutant showed an increased frequency of X-ray-induced chromosomal aberrations in the G1 and G2 stages of the cell cycle compared to wild-type frequencies. There was no increase in sister chromatid exchange levels. All of these observations in Ara-CR213 are very similar to those made with AT cells in our and other laboratories. Even more important, complementation analysis of Ara-CR213 x AT hybrid cells indicated that the gene responsible for X-ray sensitivity of AT is also mutated in Ara-CR213 cells. Thus, Ara-CR213 appears to have a mutant phenotype and probably genotype that is very similar to, if not exactly the same as, those of AT. This makes it quite different from other X-ray-sensitive cells that have been isolated in other laboratories.

The human shuttle plasmid pZ189, containing the Escherichia coli supF gene as the mutational target, was irradiated in vitro with 210Po alpha particles and transfected into human lymphoblastoid cells. Plasmids which were replicated in human cells were recovered and those containing mutant supF genes were isolated by phenotypic screening in E. coli. The mutations were characterized by sequencing the tRNA gene. The mutant frequency increased linearly with the alpha-particle dose and, at 259 Gy, it was 16 times (0.29%) that observed in unirradiated controls (0.018%). The distribution of alpha-particle-induced point mutations was highly nonrandom and similar to that observed in the unirradiated or X-irradiated plasmid DNAs. The majority of the mutations were G.C----A.T transitions and occurred selectively at most 5'-TC (3'-AG) and 5'-CC (3'-GG) sequences. For the unirradiated control DNA, these mutations at C's (G's) were preferentially located in the nontranscribed strand, similar to the observation previously made for mutations in X-irradiated DNA. Such a strand bias was not observed for mutations in the alpha-particle-irradiated DNA. The data suggest that, although similar types of point mutations are induced in unirradiated, X-irradiated, and alpha-particle-irradiated DNAs, the mechanisms of their induction and the exact nature of the lesions involved may be quite different.

The Escherichia coli supF gene encoding the suppressor tyrosine tRNA in a human shuttle plasmid, pZ189, was used as a target for molecular analysis of X-ray-induced mutations in human lymphoblastoid cells. Following replication of the in vitro-irradiated plasmid in human cells, the mutant supF-containing molecules were cloned by phenotypic screening in E. coli and the nature of the mutations was determined by direct sequencing of the tRNA gene. At 160 Gy the mutant frequency was 13 times (0.39%) that observed in unirradiated controls (0.031%). When control plasmid was replicated directly in E. coli, the mutant frequency was 16 times less than that of the plasmid passaged through the human cells. The distribution of mutations was highly nonrandom and remarkably similar in both irradiated and control DNAs. The majority of the mutations were transitions involving G.C pairs and occurred selectively at most 5'-TC (3'-AG) sequences. These mutations at C's were preferentially distributed in the nontranscribed strand. We propose that mutations in the control plasmid result from oxidative damages that occur during and/or after its incorporation into human cells and that these damages are similar to those induced by ionizing radiation. The hot spots for mutations suggest that the proximate nucleotide sequence and the overall conformation of the target DNA are important in the production and/or processing of these damages during repair and replication.

The electroporation of restriction enzymes into mammalian cells results in DNA double-strand breaks that can lead to chromosome aberrations. Four chemicals known to interfere with cellular responses to DNA damage were investigated for their effects on chromosome aberrations induced by AluI and Sau3AI; in addition, the number of DNA double-strand breaks at various times after enzyme treatment was determined by pulsed-field gel electrophoresis (PFGE). The poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide (3AB) dramatically increased the yield of exchanges and deletions and caused a small but transitory increase in the yield of double-strand breaks induced by the enzymes. 1-beta-D-Arabinofuranosylcytosine, which can inhibit DNA repair either by direct action on DNA polymerases alpha and delta or by incorporation into DNA, potentiated aberration induction but to a lesser extent than 3AB and did not affect the amount of DNA double-strand breakage. Aphidicolin, which inhibits polymerases alpha and delta, had no effect on AluI-induced aberrations but did increase the aberration yield induced by Sau3AI. The postreplication repair inhibitor caffeine had no effect on aberration yields induced by either enzyme. Neither aphidicolin nor caffeine modulated the amount of DNA double-strand breakage as measured by PFGE. These data implicate poly(ADP-ribosyl)ation and polymerases alpha and delta as important components of the cellular processes required for the normal repair of DNA double-strand breaks with blunt or cohesive ends. Comparison of these data with the effect of inhibitors on the frequency of X-ray-induced aberrations leads us to the conclusion that X-ray-induced aberrations can result from the misjoining or nonrejoining of double-strand breaks, particularly breaks with cohesive ends, but that this process accounts for only a portion of the induced aberrations.

aRadiation Effects Research Foundation, 5-2 Hijiyama Park, Minami-ku, Hiroshima 732-0815, Japan; bBiology and Biotechnology Research Program, Lawrence Livermore National Laboratory, Livermore, California 94551; CGSF, National Research Center for Environment and Health, Institute of Radiobiology, Neuherberg, Germany; dOak Ridge Institute for Sciences and Education, Oak Ridge, Tennessee 37830; eNational Radiological Protection Board, Chilton, Didcot, Oxon, United Kingdom; fCIIT, 6 Davis Drive, Research Triangle Park, North Carolina 27709; and gRadiation Biology Center, Kyoto University, Sakyo-ku, Kyoto 606-8501, Japan

The human shuttle plasmid pZ189, containing the Escherichia coli supF gene as the mutational target, was irradiated in vitro with 210Po alpha particles and transfected into human lymphoblastoid cells. Plasmids which were replicated in human cells were recovered and those containing mutant supF genes were isolated by phenotypic screening in E. coli. The mutations were characterized by sequencing the tRNA gene. The mutant frequency increased linearly with the alpha-particle dose and, at 259 Gy, it was 16 times (0.29%) that observed in unirradiated controls (0.018%). The distribution of alpha-particle-induced point mutations was highly nonrandom and similar to that observed in the unirradiated or X-irradiated plasmid DNAs. The majority of the mutations were G.C----A.T transitions and occurred selectively at most 5'-TC (3'-AG) and 5'-CC (3'-GG) sequences. For the unirradiated control DNA, these mutations at C's (G's) were preferentially located in the nontranscribed strand, similar to the observation previously made for mutations in X-irradiated DNA. Such a strand bias was not observed for mutations in the alpha-particle-irradiated DNA. The data suggest that, although similar types of point mutations are induced in unirradiated, X-irradiated, and alpha-particle-irradiated DNAs, the mechanisms of their induction and the exact nature of the lesions involved may be quite different.

The electroporation of restriction enzymes into mammalian cells results in DNA double-strand breaks that can lead to chromosome aberrations. Four chemicals known to interfere with cellular responses to DNA damage were investigated for their effects on chromosome aberrations induced by AluI and Sau3AI; in addition, the number of DNA double-strand breaks at various times after enzyme treatment was determined by pulsed-field gel electrophoresis (PFGE). The poly(ADP-ribose) polymerase inhibitor 3-aminobenzamide (3AB) dramatically increased the yield of exchanges and deletions and caused a small but transitory increase in the yield of double-strand breaks induced by the enzymes. 1-β-D-Arabinofuranosylcytosine, which can inhibit DNA repair either by direct action on DNA polymerases α and δ or by incorporation into DNA, potentiated aberration induction but to a lesser extent than 3AB and did not affect the amount of DNA double-strand breakage. Aphidicolin, which inhibits polymerases α and δ, had no effect on Alu...

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 bemore » defined as S-phase dependent.« less

The theory of the single dilution and double dilution methods for quantitative X-ray diffraction analysis is presented, and verified experimentally for the analysis of silica in a set of seven synthetic standards. Dilution methods are a new concept in diffraction analysis, and show advantages over other methods.

ONE of the major classes of genetic damage produced by ionising radiations is heritable reciprocal translocation. In a series of earlier publications, Brewen and Preston1–3 demonstrated that each of several mammalian species had its own unique sensitivity to translocation induction in peripheral leukocytes and that human and marmoset leukocytes, although approximately equal in sensitivity, were twice as sensitive as mouse leukocytes. Brewen and Preston4 also reported that in the Chinese hamster and the mouse, the frequency of the translocation type analysed in the peripheral leukocytes was three to four times as high as the frequency of heritable translocations produced in spermatogonial stem cells and recovered in primary spermatocytes. As the mouse is the mammal currently used as the model for estimating man's genetic risk from mutagenic exposure, Brewen and Preston's earlier observations on germ cells should be extended to include a primate and man if possible. Here we present an extension of the interspecific and somatic against germ cell comparisons.

On a enregistre les potentiels electriques monoet poly-synaptiques (EPSPs) des cellules du nucleus reticularis tegmenti pontis en stimulant le nucleus interpositus (NI), le brachium conjunctivum (BC) et le pedoncule cerebelleux. Les experiences de collision ont montre que des axones des cellules NRTP activent leurs neurones via BC. Quelques potentiels postsynaptiques exitateurs induits par le nucleus interpositus ont ete inverses par un courant depolarisant applique au voisinage de la microelectrode de derivation, ce qui indique que les terminaisons synaptiques d'axones NI se trouvent pres du sommet des cellules NRTP.

There are a variety of test systems available for the analysis of chromosome aberrations following in vivo treatment. There are advantages and disadvantages to each, which means that the information obtained from any one test must be critically examined. Because of the variety of test systems employed, it is also impossible to provide a concise summary of the effect of each of the chemicals under study.

Cytogenetic analyses were performed on peripheral blood from 46 present or past residents of the area surrounding Love Canal, a former dump site for chemical wastes in Niagara Falls, NY. Participants included 17 persons in whom cytogenetic analyses had been performed in 1980 and 29 persons who had been living in 1978 in seven homes that directly adjoined the canal and in which environmental tests showed elevated levels of chemicals spreading from the canal. Frequencies of chromosomal aberrations and of sister chromatid exchange (SCE) did not differ significantly from control levels. For all participants, cigarette smoking was associated with an increase in sister chromatid exchange frequency.

We have previously reported on a cytogenetic-epidemiological study of chromosomal aberration and sister-chromatid exchange (SCE) frequencies in peripheral blood lymphocytes (PBL) of a cohort of 353 healthy employees of the Brookhaven National Laboratory (Bender et al., 1988). This sample has now been increased in order to extend the age range represented and, incidentally, the representation of non-white subjects. In total, the data now include chromosomal aberration information from 108,950 cells and SCE information from 25,397 cells from 613 samples from 493 subjects. Neither the mean frequencies of any of the chromosomal aberration types nor the mean frequency of SCE have changed notably through the addition of the new subjects and samples. The mean age at sampling of the population is now 43.1 years with a range of from 1.1 to 83.7 years. However, we still find no significant relationship of the frequency of any conventional aberration category to age with the single exception of the dicentric chromosome, which now shows a positive regression (p = 0.001). The raw mean SCE frequencies show a statistically significant increase with subject age, but when cigarette smoking status is taken into account, no significant age relationship is found. As with the earlier samples, neither aberration nor SCE frequencies was influenced by race. Mean SCE frequencies, measured in non-smokers, were about 5% higher in females than males. Only one aberration category, "supernumerary acentric fragment", was significantly related to sex. This "aberration", known to constitute early centromere separation of an X chromosome, is much more common in females than in males and, in the females, increases significantly with increasing subject age.

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.

The dose-response curve for reciprocal translocations induced by X-rays in spermatogonial stem cells, and observed in primary spermatocytes of mice, is "hump-shaped", with a maximum yield at about 600 R. To test the hypothesis that the decrease in yield with increasing dose above 600 R is a consequence of the different sensitivities of cells in different stages of the cell cycle to both cell killing and chromosome aberration induction, several fractionation experiments were carried out. A total dose of 2800 R was given in repeated doses of 400 R, separated by 8-week intervals. The yield of translocations is that expected for additivity; for example, the yield at 1600 R is approximately equal to that for four separate 400-R doses. When a total dose (500 R) which gives a translocation yield on the ascending part of the dose-response curve is given as two equal fractions separated by intervals of 30, 90, or 150 min, the translocation yield decreases with increasing interval. However, when a total dose (1000 R) which would give a translocation yield on the descending part of the dose-response curve is given in two equal fractions separated by intervals of from 30 min to 6 weeks, the response is different; the translocation yield increases with intervals up to 18 h, then decreases with intervals up to 4 weeks, and finally increases again to a yield equal to additivity with an interval of 6 weeks. These changes in translocation yield with changes in interval between the two doses are explained in terms of the differential sensitivity of cells to killing and aberration induction in the different phases of the cell cycle, and by assuming that the cells surviving the first dose and repopulating the testis have different cycle characteristics from normal cells.

The thymidine (dThd) analog 5-bromodeoxyuridine (BrdUrd) is widely used in studies of the induction of sister chromatid exchanges (SCEs), since growth in the presence of BrdUrd allows the subsequent differential staining of the chromosomes through the fluorescence-plus-Giemsa (FPG) method. However, the analog itself induces SCEs, an aspect of its use which is often not considered. We have studied the induction of SCE by BrdUrd and a second dThd analog 5-chlorodeoxyuridine (CldUrd). Growth of Chinese hamster ovary (CHO) cells for 2 rounds of replication in the presence of different concentrations of either analog results in increasing results in increasing SCE frequencies which are linearly proportional to the degree of analog substitution for dThd in the DNA. However, CldUrd causes 3 to 5 times the number of SCEs found with BrdUrd, at equivalent substitution for dThd. With both analogs the increase in SCE frequency is due to the replication of the analog-substituted DNA and not to the incorporation of analog into nascent DNA. This induction of SCE can be considered at the level of a single strand of DNA since the replication of bifilarly substituted DNA results in twice the number of SCEs that are induced by the replication of unifilarly substituted DNA.

The induction of myeloid leukemia following fission neutron irradiation was examined over the 0-80 rad dose range. Over this dose range the dose response could be described by the linear regression equation: y = 0.94 + 0.18X. A comparison of these data with data obtained following gamma irradiation from this study and a previous study indicated that the relative biological effectiveness for myeloid leukemia induction was 2.8. These results appear to be compatible with those reported by other investigators.

The induction of chromosome aberrations, sister chromatid exchanges (SCE), cytotoxicity, and 6-thioguanine-resistant mutation by neocarzinostatin (NCS) in Chinese hamster ovary cells was analyzed. It was observed that within the same concentration range of 0.01-0.1 mu/ml NCS, the drug induced a significant increase in all analyzed end-points except in SCE frequencies. There was no increase in SCE frequencies even when the cells were treated at the G1/S border in the first cell cycle and when aberrations were observed in the same cell showing a second cycle differential staining pattern. Our study indicates that the cellular damage induced by NCS leads to expression in chromosome aberrations, cytotoxicity, and mutagenicity but not in sister chromatid exchanges.