Dependence of the Mutation Spectrum in a Shuttle Plasmid Replicated in Human Lymphoblasts on Dose of Gamma Radiation

Summary

The frequencies and types of mutations induced in the target gene, supF-tRNA, of the shuttle vector pZ189 were analysed following the replication of the gamma-irradiated plasmid in the human lymphoblastoid cell line, GM606. The mutation frequency measured in progeny of unirradiated pZ189 was 1·02 × 10^?4, increasing to 17·5 × 10^?4 at 1000 cGy, and to 63·4 × 10^?4 at 5000 cGy, approximately 17- and 62-fold over background levels, respectively. Simultaneously, the number of plasmids capable of replicating in Escherichia coli decreased with increasing radiation dose to 4% of the control value at 5000 cGy. Electrophoresis of the irradiated DNA showed a correlation between increases in mutation frequency and decreases in plasmid survival, and the formation of open-circular and linear DNA. The majority of the spontaneous (69·8%) and induced mutations (85·7% at 1000 and 79·4% at 5000 cGy) were base substitutions and were generally of similar types among all groups. However, changes at 2500 (12·7%) and 5000 cGy (13·2%) involving A:T base pairs were greater than those in unirradiated controls (3·4%) or those at 1000 cGy (2·0%). This increase in A:T base pair mutations could be a result of reduced repair fidelity when the DNA is extensively damaged by high doses of ionizing radiation.     

Mutations caused by gamma-radiation-induced double-strand breaks in a shuttle plasmid replicated in human lymphoblasts.

The mutagenicity of open-circular DNA (containing base damage and single-strand breaks) and linear DNA (containing base damage, single-strand breaks, and one double-strand break) produced in vitro by gamma-irradiation of shuttle vector pZ189, was analysed after the plasmid's repair and replication in the human lymphoblast line, GM606. By comparing the survival, mutation frequency, and types of mutations in descendants from the two DNA forms, the effects of the double-strand break were determined. The percentage of viable plasmids from linear DNA was two-fold lower than that from open-circular DNA, 7.8 versus 14.0 (compared with unirradiated, control DNA). The mutation frequency in progenies of the open-circular plasmid was 4.2 +/- 1.7 x 10(-3), compared with 7.8 +/- 0.1 x 10(-3) in progenies of the linear DNA, again, nearly a two-fold difference. Approximately 59% of the mutations from the linear DNA were deletions and 34% were base substitutions. In contrast, only 13% of mutations from open-circular DNA were deletions, but 87% were base substitutions. All recoverable deletions were small, ranging from 1 to 205 base pairs, and the majority contained direct repeats at the deletion junctions, indicating non-homologous recombinations. Thus, mutations found among descendants from the linear and open-circular DNAs were qualitatively similar but quantitatively different. The data suggests that producing one double-strand break in DNA by ionizing radiation causes a two-fold increase in both lethality and mutation frequency.     

Mutagenic effects of 5-formyluracil on a plasmid vector during replication in Escherichia coli

Purpose : 5-Formyluracil (5-foU) is a major derivative of thymine produced in DNA by ionizing radiation and various chemical oxidants. It has been previously shown that 5-foU in template DNA directs misincorporation of nucleotides by DNA polymerases during in vitro DNA synthesis. The present experiments were designed to understand the biological effects of 5-foU in vivo. Materials and methods : The modified base was incorporated site-specifically into the recognition site of restriction endonuclease Sal I (5'-GTCGAC) or Afl II (5'-CTTAAG) in vector plasmid pSVK3 and introduced the plasmid into Escherichia coli. Results : When the plasmids were replicated in E. coli, 5-foU caused mutations at the target sites. The induced mutation frequencies were 0.038-0.049%. Sequence analysis revealed that 5-foU preferentially caused T:A →C:G and T:A →A:T base substitutions and -1 deletions at the 5-foU site. 5-FoU also caused mutations at sites near the 5-foU. The alkA mutation did not affect the frequency of mutations in 5-foU-containing plasmids. Conclusions : The present experiments demonstrated that 5-formyluracil in DNA caused mutations in E. coli.     

Mutagenic effects of 5-formyluracil on a plasmid vector during replication in Escherichia coli

PURPOSE: 5-Formyluracil (5-foU) is a major derivative of thymine produced in DNA by ionizing radiation and various chemical oxidants. It has been previously shown that 5-foU in template DNA directs misincorporation of nucleotides by DNA polymerases during in vitro DNA synthesis. The present experiments were designed to understand the biological effects of5-foU in vivo. MATERIALS AND METHODS: The modified base was incorporated site-specifically into the recognition site of restriction endonuclease SalI (5'-GTCGAC) or AflII (5'-CTTAAG) in vector plasmid pSVK3 and introduced the plasmid into Escherichia coli. RESULTS: When the plasmids were replicated in E. coli, 5-foU caused mutations at the target sites. The induced mutation frequencies were 0.038-0.049%. Sequence analysis revealed that 5-foU preferentially caused T:A-->C:G and T:A-->A:T base substitutions and -1 deletions at the 5-foU site. 5-FoU also caused mutations at sites near the 5-foU. The alkA mutation did not affect the frequency of mutations in 5-foU-containing plasmids. CONCLUSIONS: The present experiments demonstrated that 5-formyluracil in DNA caused mutations in E. coli.     

Nucleotide excision repair proteins and their importance for radiation‐enhanced transfection

Purpose: Irradiated cells transfect more efficiently than unirradiated cells because of a radiation‐induced increase in plasmid integration. However, the molecular mechanism is unclear. Because of recent observations that nucleotide excision repair (NER) proteins can be involved in certain types of recombination in yeast, it was hypothesized that NER proteins might play a role in this radiation‐enhanced integration.

Materials and methods: Hamster and human cells with inactivating mutations in NER genes were irradiated at doses from 0 to 6?Gy and then immediately transfected with a linearized selectable marker plasmid. Transfection‐enhancement ratios (TERs) were calculated as the ratio of the number of drug‐resistant colonies in unirradiated cells to the number of transfectants in irradiated cells, corrected for cytotoxicity from radiation.

Results: Transfection into unirradiated rodent cells was unaffected by NER mutation status. Transfection into unirradiated human cells, however, was increased by NER mutation. The TERs were 5 and 100 for CHO and primary human fibroblasts, respectively, after exposure of the cells to 6?Gy. Mutations in ERCC1, XPA, XPB, XPC, XPF, XPG and CSB dramatically reduced TER. Mutations in ERCC1, XPC, XPF, XPG and CSB suppressed transfection so that the TER was significantly below 1.

Conclusions: The mechanism of radiation‐enhanced plasmid integration was distinct from that of plasmid integration in unirradiated cells, and NER gene products were critical for enhanced integration to occur.     

Dose-response relationship for radiation-induced mutations at micro- and minisatellite loci in human somatic cells in culture

PURPOSE: The study was designed to determine the dose-response relationship for radiation induction of mutations at mini- and microsatellite loci in human somatic cells. Mutations induced by graded doses of gamma-irradiation were quantified by screening clones derived from single irradiated cells for micro- and minisatellite alterations following irradiation with 1, 2 or 3 Gy. MATERIALS AND METHODS: After irradiation, the moderately radioresistant glioma cell line UVW was seeded at low density into Petri dishes to allow formation of discrete colonies, 100 of which were examined at each dose. All the cells within a colony were presumed to have arisen from a single irradiated cell. Radiation-induced microsatellite alterations were determined at 16 different loci, by PCR amplification and visualization on polyacrylamide gels. Minisatellite alterations were identified at four different minisatellite loci by restriction enzyme digestion and Southern blotting. RESULTS: A dose-response curve for mutation frequency was obtained by analysis of 100 clones, yielding a minisatellite mutation rate of 5.5x10(-3) mutations/locus/Gy/cell and a microsatellite mutation rate of 8.75x10(-4) mutations/locus/ Gy/cell. At microsatellite loci, alterations were predominantly simple loss or gain of repeat units and loss of heterozygosity (LOH). The mutations in minisatellite loci resulted predominantly in LOH and variation in repeat number. The background instability at each locus was determined by analysis of non-irradiated clones. Only 2% and 1% of the micro-and minisatellite loci respectively showed altered bands. CONCLUSIONS: This is the first report of a dose-response relationship for radiation-induced micro- and minisatellite mutations in human somatic cells. Described is a sensitive method for analysis of low-dose radiation mutagenesis in somatic cells that may prove to be a useful tool for radiation protection and dosimetry.     

One-electron oxidation of plasmid DNA by selenium(V) species

PURPOSE: To employ the gamma-radiation-generated selenium(V) one-electron-oxidizing agent SeO3*- for the preparation of guanyl radicals in plasmid DNA, and to compare the behaviour of this reagent with that of other similarly reactive oxidant species. MATERIALS AND METHODS: Plasmid DNA in aerobic aqueous solution was irradiated with 137Cs gamma-rays (662 keV). The solutions also contained up to 4x10(-2) mol x dm(-3) sodium selenate (Na2SeO4) and/or up to 10(-1) mol x dm(-3) sodium biselenite (NaHSeO3), as well as auxiliary scavengers such as DMSO or glycerol. In some cases, reducing agents such as ferrocyanide were also present. After irradiation, the plasmid was incubated with the Escherichia coli base excision-repair endonuclease formamidopyrimidine-DNA N-glycosylase (FPG). These treatments produced strand breaks in the plasmid. The yields of these strand breaks were quantified by agarose gel electrophoresis. RESULTS: In general, gamma-irradiation produced single-strand breaks (SSB) in plasmid DNA. Subsequent incubation with the endonuclease FPG increased the SSB yield by a factor of 2-100-fold. The smallest effects of FPG were observed when only DMSO or glycerol were present during irradiation. FPG incubation produced significantly larger increases in the SSB yield after gamma-irradiation in the additional presence of selenate and/or biselenite. The largest effect of FPG was observed after gamma-irradiation in the presence of 10(-2) mol x dm(-3) sodium selenate and 10(-1) mol x dm(-3) glycerol. This was indicative of extensive oxidative damage to the plasmid under these conditions and provided evidence for guanine oxidation mediated by SeO3*-. The large effect of FPG was strongly attenuated by the addition of reducing agents such as ferrocyanide. The observations suggest that these reducing agents exert their effects through the reduction of an intermediate guanyl radical. CONCLUSION: By comparing the yields of breaks produced after gamma-irradiation under a range of conditions, it is possible to formulate a reaction scheme that describes the chemical reactions responsible for the formation of strand breaks and FPG-sensitive sites. By applying this scheme to the data, we can quantify rate constants for the reduction of DNA guanyl radicals by reducing agents. This reaction is of particular interest to radiation biology because it is the equivalent of the repair of DNA damage by the direct effect of ionizing radiation.     

Nucleotide excision repair proteins and their importance for radiation-enhanced transfection

PURPOSE: Irradiated cells transfect more efficiently than unirradiated cells because of a radiation-induced increase in plasmid integration. However, the molecular mechanism is unclear. Because of recent observations that nucleotide excision repair (NER) proteins can be involved in certain types of recombination in yeast, it was hypothesized that NER proteins might play a role in this radiation-enhanced integration. MATERIALS AND METHODS: Hamster and human cells with inactivating mutations in NER genes were irradiated at doses from 0 to 6 Gy and then immediately transfected with a linearized selectable marker plasmid. Transfection-enhancement ratios (TERs) were calculated as the ratio of the number of drug-resistant colonies in unirradiated cells to the number of transfectants in irradiated cells, corrected for cytotoxicity from radiation. RESULTS: Transfection into unirradiated rodent cells was unaffected by NER mutation status. Transfection into unirradiated human cells, however, was increased by NER mutation. The TERs were 5 and 100 for CHO and primary human fibroblasts, respectively, after exposure of the cells to 6 Gy. Mutations in ERCC1, XPA, XPB, XPC, XPF, XPG and CSB dramatically reduced TER. Mutations in ERCC1, XPC, XPF, XPG and CSB suppressed transfection so that the TER was significantly below 1. CONCLUSIONS: The mechanism of radiation-enhanced plasmid integration was distinct from that of plasmid integration in unirradiated cells, and NER gene products were critical for enhanced integration to occur.     

Therapeutic external irradiation in women of reproductive age: risk estimation of hereditary effects

Exposure of women of childbearing age to ionizing radiation may result in induction of genetic disorders in future generations. This study aims to estimate the risk of hereditary effects attributable to therapeutic external irradiation in women. An anthropomorphic phantom was used to simulate radiotherapy in female patients and ovarian dose was measured for irradiation of brain, breast and lung cancer, and for treatment of Hodgkin's disease. These malignancies are among the most common tumours presenting in women of reproductive age. Dose measurements were undertaken using thermoluminescent dosemeters and all exposures were made with 6 MV X-ray beams. The dose to ovaries was found to be 2-3 cGy, 8-11 cGy and 11-15 cGy depending on the distance from the primary irradiation field during radiotherapy of brain, breast and lung cancer, respectively. The corresponding ovarian dose resulting from treatment of supradiaphragmatic and infradiaphragmatic Hodgkin's disease was 18-25 cGy and 128-356 cGy, respectively. A small excess risk of genetic diseases of (1-15) x 10(-4) was estimated for radiotherapy above the diaphragm. Pelvic irradiation resulted in an increased risk of hereditary effects of (77-214) x 10(-4).     

Reaction of guanyl radicals in plasmid DNA with biological reductants: chemical repair of DNA damage produced by the direct effect of ionizing radiation

Purpose : It has been previously argued that the use of the one-electron oxidants (SCN) 2 ?- and Br 2 ?- with plasmid DNA leads to the formation of DNA guanyl radicals. These guanyl radical species are intermediates in the DNA damage produced by processes such as photo-ionization and ionizing irradiation. The present paper evaluates the use of thallium(II) ions (Tl II OH +) as the one-electron oxidant, and also determines rate constants for the reduction (repair) of guanyl radicals in plasmid DNA by a variety of reducing agents including the biologically important compounds ascorbate and glutathione. Materials and methods : Aqueous solutions of plasmid DNA containing 10 -3 mol dm -3 thiocyanate or thallous ions and a reducing agent (azide, nitrite, ferrocyanide, hexachloroiridate(III), iodide, ascorbate, glutathione, glutathione disulphide, methionine, tyrosine, 5-hydroxyindole-3-acetic acid, 10 -7 -10 -4 mol dm -3) were irradiated with 137 Cs γ-rays (662 keV). After irradiation, the plasmid was incubated with the E. coli base excision repair endonuclease formamidopyrimidine-DNA N -glycosylase (FPG). Strand break yields after incubation were quantified by means of agarose gel electrophoresis. Results : High yields of FPG-sensitive sites produced by the oxidants (SCN)2 ?- and Tl II OH + were strongly attenuated by the presence of the reducing agents. Conclusions : From the results, it is possible to arrive at estimates of the rate constants for the reduction of the DNA guanyl radical by the reducing agents. Values lie in the range 10 4 -10 7 dm 3 mol -1 s -1. Using the values for ascorbate and glutathione, it is possible to estimate an upper limit on the order of milliseconds for the lifetime of DNA guanyl radicals under cellular conditions. The implication is that there may well be a significant chemical repair of DNA base damage by the direct effect of ionizing radiation.     

Reaction of guanyl radicals in plasmid DNA with biological reductants: chemical repair of DNA damage produced by the direct effect of ionizing radiation.

PURPOSE: It has been previously argued that the use of the one-electron oxidants (SCN)2(*-) and Br2(*-) with plasmid DNA leads to the formation of DNA guanyl radicals. These guanyl radical species are intermediates in the DNA damage produced by processes such as photo-ionization and ionizing irradiation. The present paper evaluates the use of thallium(II) ions (Tl(II)OH(+)) as the one-electron oxidant, and also determines rate constants for the reduction (repair) of guanyl radicals in plasmid DNA by a variety of reducing agents including the biologically important compounds ascorbate and glutathione. MATERIALS AND METHODS: Aqueous solutions of plasmid DNA containing 10(-3) mol dm(-3) thiocyanate or thallous ions and a reducing agent (azide, nitrite, ferrocyanide, hexachloroiridate(III), iodide, ascorbate, glutathione, glutathione disulphide, methionine, tyrosine, 5-hydroxyindole-3-acetic acid, 10(-7)-10(-4) mol dm(-3)) were irradiated with 137Cs gamma-rays (662 keV). After irradiation, the plasmid was incubated with the E. coli base excision repair endonuclease formamidopyrimidine-DNA N-glycosylase (FPG). Strand break yields after incubation were quantified by means of agarose gel electrophoresis. RESULTS: High yields of FPG-sensitive sites produced by the oxidants (SCN)2(*-) and Tl(II)OH(+) were strongly attenuated by the presence of the reducing agents. CONCLUSIONS: From the results, it is possible to arrive at estimates of the rate constants for the reduction of the DNA guanyl radical by the reducing agents. Values lie in the range 10(4)-10(7) dm(3) mol(-1) s(-1). Using the values for ascorbate and glutathione, it is possible to estimate an upper limit on the order of milliseconds for the lifetime of DNA guanyl radicals under cellular conditions. The implication is that there may well be a significant chemical repair of DNA base damage by the direct effect of ionizing radiation.     

Redox reactivity of guanyl radicals in plasmid DNA

Purpose : It has been previously argued that γ-irradiation of plasmid DNA in the presence of thiocyanate ions produces products recognized by the E. coli base excision-repair endonuclease formamidopyrimidine-DNA N -glycosylase (FPG), and there that derive from an intermediate guanyl radical species. The wish was to characterize the reactivity of this intermediate with reducing agents. Materials and methods : Aqueous solutions of plasmid DNA containing either bromide or thiocyanate (10 -3 to 10 -1 mol dm -3) and also one of six other additives (azide, ferrocyanide, iodide, nitrite, promethazine, tryptophan, 10 -7 to 10 -3 mol dm -3) were subjected to 137 Cs γ-irradiation (662 keV). After irradiation, the plasmid was incubated with FPG. Strand break yields before and after incubation were determined by agarose gel electrophoresis under neutral conditions. Results : The very high yields of FPG-sensitive sites in the presence of SCN - or Br - decreased significantly with increasing concentrations of all of the six additives, with promethazine and tryptophan being the most efficient additives, and azide and iodide the least. Conclusions : From the results it is possible to estimate values of the rate constants for the reduction of the DNA guanyl radical (5x10 5, 2x10 5, 10 7 and 10 7 dm 3 mol -1 s -1 for ferrocyanide, nitrite, promethazine and tryptophan respectively).     

Minisatellite and Hprt mutations in V79 cells irradiated with helium ions and gamma rays

PURPOSE: To evaluate and compare cytotoxic and mutational effects of graded doses of gamma-rays and 4He++ ions at different LET values (nominally 80 and 123 keV/microm) in V79 cells. MATERIALS AND METHODS: 4He++ ion beams at 80 and 123 keV/microm were supplied by the 7 MV Van de Graaff CN accelerator of the INFN-LNL in the dose range 0.3 2.4 Gy at a dose rate of 1 Gy/min. Gamma-irradiation was performed by the 60Co 'gamma beam' of CNR-FRAE (at the INFN-LNL) in the dose range 0.5 6.0 Gy at a dose rate of 1 Gy/min. After irradiation, the cells were seeded to measure surviving fraction (SF) and mutant frequency (MF) at the Hprt locus on the basis of 6-thioguanine resistance. Alterations at minisatellite sequences (MS) of clones derived from irradiated and unirradiated cells were detected by Southern blot analysis using a multi-locus probe (DNA fingerprinting). RESULTS: Survival data from 4He++ irradiation at two LET values (80 and 123 keV/microm) yielded similar results: alpha = (1.08 +/- 0.04)/Gy and (0.90 +/- 0.03)/Gy, respectively. The best fit for mutant induction at the Hprt locus after 80keV/microm 4He++ was a linear function of the dose in the dose-interval 0-1.5 Gy: alpha= (47.77 +/- 16.01) x 10(-6)/Gy. The best fit for mutant induction after 123 keV/microm 4He++ in the dose-interval 0-1.2 Gv was a linear-quadratic function: alpha=(86.01 +/- 13.80) x 10(-6)/Gy; beta = (42.87 +/- 11.03) x 10(-6)/Gy2. For gamma-irradiation, the best fit of Hprt mutation data gave: alpha = (4.14+2.67)x 10(-6)/Gy: beta = (0.63 +/- 0.86) x 10(-6)/Gy2. The best fitting of MS alteration data with linear-quadratic or linear relationships gave: for gamma-rays, alpha = 0.56 mutants/Gy and beta = 0.52 mutants/Gy2; for 80 keV/microm 4He++, alpha = 3.70 mutants/Gy and beta = 9.00 mutants/Gy2; for 123keV/microm 4He++, alpha = 4.36 mutants/Gy. CONCLUSIONS: The results reported here confirmed the higher cytotoxic and mutagenic effects of helium ions in comparison with gamma-irradiation and the ability of DNA fingerprint analysis to investigate DNA damage induced by different ionizing radiations. The results of the mutagenic effects measured by the two tests are in agreement.     

The DNA Content of the Small Intestine as a Quantitative Measure of Damage and Recovery after Whole Body Irradiation

Summary

(1) The total amount of deoxyribonucleic acid (DNA) in the small intestine was used as a measure of cell population in order to follow the course of radiation damage in mice and rats. Changes in total DNA-P were proportionately larger and intrinsically less variable than changes in fresh weight of the intestine.

(2) In unirradiated control mice intestinal DNA-P concentration was constant but intestinal weight and total intestinal DNA-P were linearly dependent on body weight.

(3) Single doses of whole body irradiation given to mice caused a fall in total intestinal DNA-P which began 12 hours later. Except after 25 000 r which had less effect than 5000 r, the larger the dose of radiation the greater the maximum depression in DNA-P and the later the time at which recovery began. The course of recovery was exponential after 1000 r but not after 750 r.

(4) In both rats and mice β-mercaptoethylamine reduced the damaging effect of a dose of radiation but did not modify the course of recovery.

(5) Chronic irradiation of mice at levels sufficient to reduce body weight slightly caused a loss in total intestinal DNA-P. The loss was probably, but not certainly, greater than that expected from the loss in body weight alone.     

Expression of lethal mutations in progeny of irradiated mammalian cells

The cloning efficiency of the progeny of CHO and BALB/3T3 cells surviving acute exposures of 100-1000 cGy of X-rays was examined by three different experimental protocols. A dose-dependent decrease in cloning efficiency was observed with both cell types up to 13-15 population doublings after exposure. This decrease persisted for longer times after high radiation doses; for example, the cloning efficiency of the progeny of 3T3 cells 28 population doublings after irradiation with 1000 cGy was 44 per cent of that in parallel non-irradiated controls. Confluent holding under conditions which allowed the repair of potentially lethal damage had no effect on this phenomenon. These results are consistent with the hypothesis that large numbers of lethal mutations may be expressed among the progeny of surviving cells for many generations after irradiation.     

Redox reactivity of guanyl radicals in plasmid DNA

PURPOSE: It has been previously argued that gamma-irradiation of plasmid DNA in the presence of thiocyanate ions produces products recognized by the E. coli base excision-repair endonuclease formamidopyrimidine-DNA N-glycosylase (FPG), and there that derive from an intermediate guanyl radical species. The wish was to characterize the reactivity of this intermediate with reducing agents. MATERIALS AND METHODS: Aqueous solutions of plasmid DNA containing either bromide or thiocyanate (10(-3) to 10(-1) mol dm(-3)) and also one of six other additives (azide, ferrocyanide, iodide, nitrite, promethazine, tryptophan, 10(-7) to 10(-3) mol dm(-3)) were subjected to 137Cs gamma-irradiation (662 keV). After irradiation, the plasmid was incubated with FPG. Strand break yields before and after incubation were determined by agarose gel electrophoresis under neutral conditions. RESULTS: The very high yields of FPG-sensitive sites in the presence of SCN- or Br- decreased significantly with increasing concentrations of all of the six additives, with promethazine and tryptophan being the most efficient additives, and azide and iodide the least. CONCLUSIONS: From the results it is possible to estimate values of the rate constants for the reduction of the DNA guanyl radical (5 x 10(5), 2 x 10(5), 10(7) and 10(7) dm3 mol(-1) s(-1) for ferrocyanide, nitrite, promethazine and tryptophan respectively).     

Critical energies for SSB and DSB induction in plasmid DNA by low-energy photons: action spectra for strand-break induction in plasmid DNA irradiated in vacuum

PURPOSE: To measure action spectra for the induction of single-strand breaks (SSB) and double-strand breaks (DSB) in plasmid DNA by low-energy photons and provide estimates for the energy dependence of strand-break formation important for track-structure simulations of DNA damage. MATERIALS AND METHODS: Plasmid pMSG-CAT was irradiated as a monolayer, under vacuum, with 7 150eV photons produced by a synchrotron source. Yields of SSB and DSB were determined by the separation of the three plasmid forms by gel electrophoresis. RESULTS: The yields of SSB per incident photon increased from 1.4x 10(-15) SSB per plasmid per photon/cm2 at 7eV to 7.5 x 10(-14) SSB per plasmid per photon/cm2 at 150 eV. Direct induction of DSB was also detected increasing from 3.4 x 10(-17) DSB per plasmid per photon/cm2 at 7eV to 4.1 x 10(-15) DSB per plasmid per photon/cm2 at 150eV. When the absorption cross-section of the DNA was considered, the quantum efficiency for break formation increased over the energy range studied. Over the entire energy range, the ratio of SSB to DSB remained constant. CONCLUSIONS: These studies provide evidence for the ability of photons as low as 7 eV to induce both SSB and DSB. The common action spectrum for both lesions suggests that they derive from the same initial photoproducts under conditions where the DNA is irradiated in vacuum and a predominantly direct effect is being observed. The spectral and dose-effect behaviour indicates that DSB are induced predominantly by single-event processes in the energy range covered.     

Ultraviolet difference spectral studies in the gamma radiolysis of DNA and model compounds. I. Aqueous solutions of DNA bases

Ultraviolet difference spectra of gamma-irradiated, air-saturated aqueous solutions of DNA bases vs. unirradiated solutions of the same bases are shown to be a very sensitive supplemental tool with which to investigate the yields, postirradiation kinetics, and general nature of DNA base radiation products. Irradiated pyrimidines yield difference spectra which are approximately negative mirror-images of the base absorption spectra in the near-UV, indicating loss of ring conjugation. Difference spectra of irradiated purines yield a more complex pattern containing a positive long-wavelength peak, interpreted as radiation-induced extension of conjugation of the pi electron system beyond that of the unirradiated purine. On the basis of the spectroscopic evidence from these studies, 8-hydroxyguanine appears to be the dominant UV-absorbing radiation product in air-saturated guanine solutions with a G-value of 0.3 molec (100 eV)-1. Difference spectral studies provide isosbestic points which can be used in testing proposed radiation products and their yields. Such spectral studies are a rapid, non-invasive, supplemental tool which can be employed in conjunction with other analytical techniques in radiation-chemical studies, and which is one of the few tools able to detect short-lived chemical intermediates observed in oxygenated solutions of irradiated purines.     

Radiation dose to the fetus from body MDCT during early gestation

OBJECTIVE: The purpose of our study was to determine radiation dose to the fetus at early gestation when contemporary MDCT scanners are used for common clinical indications. MATERIALS AND METHODS: Anthropomorphic phantoms were constructed to reflect a pregnant woman. Thermoluminescence dosimeters (TLDs) and metal oxide semiconductor field effect transistor (MOSFET) detectors were placed in appropriate locations to determine real-time radiation exposure to the fetus at 0 and 3 months' gestation. Imaging was performed on a 16-MDCT scanner using current institutional CT protocols: renal stone (140 kVp, 160 mA, rotation time of 0.5 sec, 16 x 0.625 mm), appendix (140 kVp, 340 mA, rotation time of 0.5 sec, 16 x 0.625 mm), and pulmonary embolus (140 kVp, 380 mA, rotation time of 0.8 sec, 16 x 1.25 mm). RESULTS: The radiation dose to the fetus at 0 and 3 months, respectively, was as follows: renal stone protocol, 0.8-1.2 and 0.4-0.7 cGy; appendix protocol, 1.52-1.68 and 2-4 cGy; and pulmonary embolus protocol, 0.024-0.047 and 0.061-0.066 cGy. CONCLUSION: Radiation doses to the fetus from institutional MDCT protocols that may be used during pregnancy (for pulmonary embolus, appendicitis, and renal colic) are below the level thought to induce neurologic detriment to the fetus. Imaging the mother for appendicitis theoretically may double the fetal risk for developing a childhood cancer. Radiation doses to the fetus from pulmonary embolus chest CT angiography are of the same magnitude as ventilation-perfusion (V/Q) scanning.     

Effects of radiation therapy on oesophageal transit in patients with inner quadrant breast tumour

OBJECTIVES: The aim of this study was to evaluate quantitatively the effect of low doses of radiation therapy on the oesophageal transit in patients with inner quadrant breast carcinoma. METHODS: Eighteen female patients with locally advanced inner quadrant breast tumour were included in this study. A total dose of 5000 cGy in 25 fractions of 200 cGy was applied from four different treatment portals to all patients. Oesophageal motility was evaluated before and immediately after radiotherapy using oesophageal scintigraphy. The oesophageal transit times (ETTs) for the upper one-third, the lower two-thirds portion and the whole oesophagus were calculated. RESULTS: The upper one-third portion of the oesophagus received a mean dose of 600 cGy and the lower two-thirds portion received a mean dose of 1530 cGy as a result of 5000 cGy dose application. All of the patients developed grade 1 oesophageal toxicity. Post-radiation therapy ETT values were significantly higher compared to pre-radiation therapy ETT values (P<0.001). Before irradiation, ETT values for the upper one-third, distal two-thirds of the oesophagus and the whole oesophagus were 4.77+/-1.08, 11.22+/-2.85 and 11.61+/-2.97 s, respectively. After irradiation, ETT values for the upper one-third, distal two-thirds of the oesophagus and the whole oesophagus were 6.92+/-2.15, 23.30+/-5.65 and 23.74+/-5.70 s, respectively. CONCLUSIONS: Oesophageal transit seems to be affected by radiation even without a clinically significant oesophageal symptom and oesophageal scintigraphy allows sensitive, non-invasive and quantifiable assessment of the oesophageal transit time.