Search for bosonic superweakly interacting massive dark matter particles with the XMASS-I detector.

Bosonic superweakly interacting massive particles (super-WIMPs) are a candidate for warm dark matter. With the absorption of such a boson by a xenon atom, these dark matter candidates would deposit an energy equivalent to their rest mass in the detector. This is the first direct detection experiment exploring the vector super-WIMPs in the mass range between 40 and 120 keV. With the use of 165.9 day of data, no significant excess above background was observed in the fiducial mass of 41 kg. The present limit for the vector super-WIMPs excludes the possibility that such particles constitute all of dark matter. The absence of a signal also provides the most stringent direct constraint on the coupling constant of pseudoscalar super-WIMPs to electrons. The unprecedented sensitivity was achieved exploiting the low background at a level 10(-4) kg-1 keVee-1 day-1 in the detector.

Somatic-cell mutation induced by UVA and monochromatic UV radiation in repair-proficient and -deficient Drosophila melanogaster.

Near-ultraviolet light (UVA: 320-400 nm) constitutes a major part of sunlight UV. It is important to know the effect of UVA on the biological activities of organisms on the earth. We have previously reported that black light induces somatic-cell mutation in Drosophila larvae. To investigate which wavelength of the UVA is responsible for the mutation we have now carried out a series of monochromatic irradiations (310, 320, 330, 340, 360, 380 and 400 nm) on Drosophila larvae, using the large spectrograph of the National Institute for Basic Biology (Okazaki National Research Institutes, Okazaki, Japan). Mutagenic activity was examined by the Drosophila wing-spot test in which we observe mutant wing hair colonies (spots) on the wings of adult flies obtained from the treated larvae. The induction of mutation was highest by irradiation at 310 nm and decreased as the wavelength became longer. Neither the 380 nor the 400 nm light was mutagenic. Excision repair is known to protect cells from UV damage. In the excision-repair-deficient Drosophila, the mutagenic response induced by 310 nm irradiation was 24-fold higher than that of the wild-type (7.2 +/- 1.5 spots/wing/kJ vs 0.3 +/- 0.08 spots/wing/kJ), and at 320 nm the difference of the response was 14-fold (0.21 vs 0.015 +/- 0.005). In the case of irradiation at 330 and 340 nm the difference of the response was only two-fold (at 330 nm, 6.9 +/- 2.9 x 10(-3) vs 3.1 +/- 1.1 x 10(-3) spots/wing/kJ; at 340 nm, 3.5 +/- 0.9 x 10(-3) vs 2.0 +/- 0.7 x 10(-3). These results suggest that the lesion caused in the larvae by 320 nm irradiation may be similar to the damage induced by 310 nm and that the lights of 330 and 340 nm may induce damage different from the lesions induced by shorter-wavelength lights.

Comparisons of spore dosimetry and spectral photometry of solar-UV radiation at four sites in Japan and Europe.

In order to develop monitoring and assessment systems of biologically effective doses of solar-UV radiation, concurrent measurements of spectral photometry and spore dosimetry were conducted in summer months at four sites in Japan and Europe. Effectiveness spectra were derived by multiplying spectral irradiance in 0.5 nm steps between 290 and 400 nm with the inactivation efficiency of the spores determined using monochromatic radiation of fine wavelength resolution. Shapes of the effectiveness spectra were very similar at the four sites exhibiting major peaks at 303.5, 305.0, 307.5 and 311.0 nm. The dose rates for spore inactivation from direct survival measurements and from calculations by the integration of the effectiveness spectra were compared for 174 data points. The ratios (observed/calculated) of the two values were concordant with a mean of 1.26 (+/- 0.24 standard deviation [SD]). The possible causes for the variations and slightly larger observed values are discussed.

Base substitution spectra of nalidixylate resistant mutations induced by monochromatic soft X and 60Co gamma-rays in Bacillus subtilis spores.

Bacillus subtilis spores were exposed to three types of photons, monochromatic soft X-rays with the energy corresponding to the absorption peak of phosphorus K-shell electron (2,153 eV) and with the slightly lower energy (2,147 eV), and 60Co gamma-rays. From the irradiated spores, 233 mutants exhibiting nalidixic acid resistance were isolated, and together with 94 spontaneous mutants, the sequence changes in the 5'-terminal region of the gyrA gene coding for DNA gyrase subunit A were determined. Among eighteen alleles of the gyrA mutations, eight were single-base substitutions, nine were tandem double-base substitutions, and one was a double substitution skipping a middle base pair. About 6% of the radiation-induced mutations were tandem double-base substitutions, whereas none was observed among the spontaneous ones. Among spontaneous mutations, A:T and G:C pairs were equally subjected to mutations, whereas the substitutions from G:C pairs and those to A:T pairs predominated among those induced with soft X-rays. The peak-energy X-rays were more effective in killing and causing mutations than the low-energy X-rays, however, there seemed no base-change events uniquely attributable to phosphorus K-shell absorption.

On the experimental distinction between ssbs and dsbs in circular DNA.

PURPOSE: To determine directly the minimal distance between two ssbs on complementary strands in circular DNA that are not observed as a dsb by electrophoresis. MATERIALS AND METHODS: 3.2 kbp DNAs with cohesive overhangs of various lengths were systematically generated by a newly devised method and electrophoresed in agarose slabs. RESULTS: At 4 degrees C, 3.2 kbp DNA with cohesive overhangs larger than 6 bp migrated as circular DNA. The minimal overhang size for the DNA to migrate as circular DNA was larger at 25 degrees C. Whether the DNA migrated as a circular or a linear molecule depended also on the nucleotide sequence of its overhangs, most notably at the minimal size. CONCLUSIONS: The minimal distance obtained in the present study agrees with the smaller values of previous indirect estimates. The dependence of the distance on experimental conditions suggests feasibility of obtaining the stagger-size distribution of radiation-induced dsbs.

Induction of unique tandem-base change mutations in bacterial spores exposed to extreme dryness.

Despite the remarkable resistance to desiccation, Bacillus subtilis spores manifest indications of DNA damage when being kept in an extremely dry environment made by high vacuum. Spores of strain TKJ3422 (uvrA10 spl-1 recA4) with triple repair defects lost colony-forming capacity dependent on the duration and strength of the exposure. Mutations to rifampicin resistance were induced in the spores of the strain HA101 with wild-type repair capability and the strain TKJ6312 (uvrA10 spl-1) with double repair defects. The majority of nalidixic acid-resistant mutations induced by the exposure to high vacuum belonged to one particular allele gyrA12 carrying a tandem-base change, 5'-CA to 5'-TT, at codon 84 of the gyrA gene coding for DNA gyrase subunit A. This allele has never been found among more than 500 mutants obtained by various treatments other than vacuum exposure. These results indicate forced dehydration of DNA in the microenvironment of the spore core causes unique damage leading to lethal and mutagenic consequences.

Single- and double-strand breaks in pBR322 plasmid DNA by monochromatic X-rays on and off the K-absorption peak of phosphorus.

Using a synchrotron irradiation system pBR322 plasmid DNA was irradiated under vacuum by monochromatic X-rays having five specific photon energies (2.147, 2.153, 2.159, 2.168 and 2.199 keV) both on and off the K-absorption peak (2.153 keV) of phosphorus. The single- and double-strand breaks (ssb and dsb) were measured as conversions of the closed circular form of DNA (form I) to open circular (form II) and linear (form III) forms respectively. Exposures to induce one strand break per molecule were lowest at the peak (2.153 keV), and highest at 2.147 keV; the ratios were 2.7 for ssb and 3.0 for dsb. The exposures for dsb were 21-26 times higher than those for ssb. When the exposures were converted to absorbed doses in grays the absorbed doses per ssb were almost independent of photon energy. This result indicates that a certain absorbed dose was necessary to induce a ssb, regardless of whether photons were absorbed by the K-shell of phosphorus or by other shells, or by other atoms. However, the absorbed dose per dsb at 2.147 keV was 1.17 times higher than that averaged over four X-ray energies above 2-153 keV, indicating that the K-shell absorption, and the subsequent Auger event, efficiently induce dsb. The results are also discussed concerning the number of photo-absorptions of the constituent atoms per DNA strand break.

Experimental correspondence between spore dosimetry and spectral photometry of solar ultraviolet radiation.

The biologically effective dose of solar UV radiation was estimated from the inactivation of UV-sensitive Bacillus subtilis spores. Two types of independent measurements were carried out concurrently at the Aerological Observatory in Tsukuba: one was the direct measurement of colony-forming survival that provided the inactivation dose per minute (ID/min) and the other was the measurement of the spectral irradiance by a Brewer spectrophotometer. To obtain the effective spectrum, the irradiance for each 1 nm wavelenght interval from 290 to 400 nm was multiplied with the efficiency for inactivation derived from the inactivation action spectrum of identically prepared spore samples. Integration of the effective spectrum provided the estimate for ID/min. The observed values of ID/min were closely concordant with the calculated values for the data obtained in four afternoons in 1993. The average ratio (+/- SD) between them was 1.24 (+/- 0.16) for 14 data points showing high inactivation rates (> 0.05 ID/min). Considering difficulties in the absolute dosimetry of UV radiation, the concordance was satisfactory and improved credibility of the two types of monitoring systems of biologically effective dose of solar UV radiation.

Radiolytic products of bromodeoxyuridine in solids by 60Co gamma-rays and monoenergetic soft x-rays at the K-absorption edge of bromine.

In order to investigate DNA damage due to Auger cascades in bromodeoxyuridine (BrdU), BrdU mixed with other nucleosides, as a model of DNA, was irradiated in solids by gamma-rays and monoenergetic x-rays at around the K-absorption edge of bromine (13.47 keV). The main products of BrdU were deoxyuridine produced through debromination, and bromouracil produced through the decomposition of a sugar group. The rates of the debromination and the nucleobases release of additives were markedly increased in the mixed sample. This observation indicated that the additives surrounding BrdU efficiently supplied protons and then decomposed. The major products by x-rays were the same as those by gamma-rays, indicating that Auger cascades in bromine atoms did not produce specific products. The production rates for all products from the mixed sample were about 2.5 times higher at 13.51 (above the K-absorption edge) keV than at 13.43-keV x-rays.

Lethal effect of K-shell absorption of intracellular phosphorus on wild-type and radiation sensitive mutants of Escherichia coli.

The present study was conducted to clarify the lethality of Auger cascades induced by the K-shell photoabsorption of phosphorus in Escherichia coli. Killing of wild-type and radiation-sensitive mutants of E. coli was examined. Three x-ray energies were chosen for irradiation; at 2.153 keV: the resonance peak of K-shell photoabsorption of phosphorus; at 2.146 and 2.160 keV: off-peak. Enhancement ratio, which was defined as the ratio of the killing sensitivity of 2.153 keV to that at 2.146 keV, were 1.32 to 1.54 for examined strains. Increment of absorbed energy calculated in entire cells for 2.153 keV radiation could not explain the degree of observed enhancement of killing. Lethality of Auger cascades depended on the killing sensitivity with x-rays which did not induce Auger cascades. The lethality for wild-type was lower than that for recombination repair-deficient mutants. It was concluded that one part of damages produced by Auger cascades was repaired in wild-type strains.

DNA damage induced by vacuum and soft X-ray photons from synchrotron radiation.

The recent development of irradiation systems using synchrotron radiation (SR) as a source is enabling researchers to obtain intense monochromatic photons having a narrow bandwidth in the vacuum-UV (VUV) and soft X-ray regions. We can thus systematically study the photon energy dependence of DNA damage formation in these energy regions. The photon energy dependence provides useful information about how energy-absorbing modes--excitations, so-called superexcitations, outer- and inner-shell ionizations--affect the type and amount of DNA damage. Furthermore, low energy electrons produced by low energy photons through photoelectric interactions are useful for studying how the electron energy affects the induction of DNA damage. A report is given on the present status of the SR irradiation systems in Japan as well as some results concerning the formation of DNA damage, in vitro and in vivo, by monochromatic photons in the VUV and soft X-ray regions.

Dithymine photodimers and photodecomposition products of thymidylyl-thymidine induced by ultraviolet radiation from 150 to 300 nm.

Solid thymidylyl-(3'-->5')-thymidine (dTpdT) was irradiated in a vacuum with monochromatic photons from 150 to 300 nm; the photoproducts were analyzed quantitatively by high-performance liquid chromatography. The results of the experiment were as follows: (1) above 210 nm the major photoproducts were three dithymine photodimers [the cis-syn and trans-syn cyclobutane thymine dimers and the thymine (6-4) photoproduct]; below 210 nm, they were three photodecomposition products (thymine, thymidine 5'-monophosphate and thymidine 3'-monophosphate). This shows that 210 nm is the wavelength at which the major photoproducts change from dithymine photodimers (far-UV type) to photodecomposition products (X-ray type). (2) The yields of the three dithymine photodimers had a similar wavelength dependence with each other: the yields had a peak at 260 nm and gradually decreased toward shorter wavelengths to 150 nm. (3) The yields of the three photodecomposition products also had a very similar wavelength dependence with each other: the yields increased exponentially with a decrease in the wavelength. (4) The average ratios of the yield of the (6-4) photoproduct to that of the cis-syn dimer were 0.30 between 170 and 220 nm, but 0.16 between 240 and 290 nm.

Single- and double-strand breaks in pBR322 DNA by vacuum-UV from 8.3 to 20.7 eV.

Solid pBR322 DNA was irradiated in a vacuum by monochromatic photons from 8.3 eV (150 nm) to 20.7 eV (60 nm), and the formation of single- and double-strand breaks (ssb and dsb) was determined using agarose gel electrophoresis. The action cross sections increased 20 times (ssb) and 43 times (dsb) from 8.3 eV to 20.7 eV; the quantum yields increased 5.4 times (ssb) and 12 times (dsb). The cross sections for dsb were 0.0059 at 8.3 eV and 0.013 at 20.7 eV of those of ssb; these values were smaller than about 0.04 for 2.1 keV photons, or about 0.18 for 60Co gamma-rays. Although vacuum UV photons were proved to induce dsb, they had a lower efficiency than when using soft X-rays or gamma-rays.

Uracil-DNA glycosylase produces excess lethal damage induced by an Auger cascade in BrdU-labelled bacteriophage T1.

T1 phages with and without 5-bromo-2'-deoxyuridine (BrdU) labelling were irradiated in solids with monochromatic X-rays at 12.40 and 13.51 keV, below and just above the K-absorption edge of bromine (13.47 keV) in vacuum and wet states. Irradiated phages were assayed on uracil-DNA glycosylase (Udg) deficient (ung-1) and sufficient (ung+) host strains of Escherichia coli, in order to investigate the nature of the lethal damage induced by Auger cascade following X-ray absorption at the K-shell of bromine as a key atom. The results were: (1) An Auger-specific enhancement (1.15) was observed only when BrdU-labelled phages were irradiated in the wet state and assayed on ung+ host cells. (2) A Udg-specific enhancement was observed only for BrdU-labelled phages, not for unlabelled phages. (3) The sensitivities of BrdU-unlabelled phages were almost the same, despite the irradiation states and strains of the host cell, indicating that this sensitivity was a common fraction of the sensitivity under all conditions. (4) The lethal damage for the T1 phage was categorized into four fractions according to the sensitivities under different conditions: the general fraction was defined as being the sensitivity of unlabelled phages (G-fraction); BrdU-specific, but unrecognizable by Udg (B-fraction); Udg specific, but not Auger-specific (U-fraction); and Auger-specific (A-fraction). (5) Although the so-called indirect action of water radicals increased only the G-fraction by about three-fold, the B- and U-fractions were not affected by any change in the irradiation states, thus indicating that these two fractions were caused by the so-called direct action.

Effects of the K-shell X-ray absorption of phosphorus on the scission of the pentadeoxythymidylic acid.

The scission of pentadeoxythymidylic acid, d(pT)5, by monochromatic soft X-rays on (2153 eV) and below (2147 eV) the K-shell absorption peak of phosphorus was studied as a model of strand breakage in DNA. Samples dried on glass plates were irradiated by monochromatic soft X-rays in vacuo, and the products were analysed by HPLC. The main products, in ascending order of the retention time, were thymine, 5'-dTMP, d(pT)2, d(pT)3, d(pT)4, d(pTp) and three unknown products (UK 1, 2 and 3), which were presumed to be d(pTpTp), d(pTpTpTp) and d(pTpTpTpTp), respectively. No difference between 2147 and 2153 eV irradiation in the nature of the induced products was detected, indicating that the K-shell absorption of phosphorus and the following Auger process produced the same types of products as those produced by photoabsorption at the other shells of phosphorus and at other atoms. The cross-sections for the induction of products at 2153 eV were 3.3-4.0 times larger than those at 2147 eV, the ratios of these values being scattered around the ratio (3.65) of absorbance of the sample between 2153 and 2147 eV. The dependence on the X-ray energy, however, almost disappeared after conversion from exposure to absorbed dose; the ratios of the G-values (number of products per 100 eV) of the products were 0.92-1.11. Photoabsorption at the K-shell of phosphorus induced products comparable or slightly less effectively than photoabsorption at the others. These results indicate that the K-shell absorption of phosphorus and the following Auger process do not have any characteristic effect on strand breakage in dry DNA, either qualitatively or quantitatively.

Inactivation action spectra of Bacillus subtilis spores with monochromatic soft X rays (0.1-0.6 nm) of synchrotron radiation.

Five types of Bacillus subtilis spores differing in DNA repair and recombinational capacities were exposed in vacuum to monochromatic soft X rays from synchrotron radiation. The inactivation rate constants were obtained from exposure-survival curves upon irradiations at 12 wavelengths in the range of 0.1000 nm (12.40 keV) to 0.6000 nm (2.066 keV). Spores of two repair-deficient strains, UVS (uvrA ssp) and UVP (uvrA ssp polA), exhibited almost equal sensitivities to those of wild-type UVR+, while those of two recombination-deficient strains, RCE (recE) and RCF (recF), exhibited higher sensitivities in the whole wavelength range. This suggested that the repair of DNA damage produced by soft X rays was dependent on the recombinational capabilities. Inactivation action spectra based on photon fluence showed that the effectiveness of the radiation increased as the wavelengths became longer. Abrupt changes in the effectiveness occurred around the wavelengths corresponding to the absorption edges of K-shell electrons of phosphorus and calcium. In both cases, the sensitivity was the highest at the wavelengths of the resonance absorption peak, the next highest at those of the higher energy, and the lowest at the lower energy. Mass energy absorption coefficients of spores were obtained from the transmission of a flake made of spores. They were used to derive inactivation action spectra based on absorbed doses. In these spectra, basal levels of the sensitivity seemed constant, and enhancements of the sensitivity were observed consistent with the absorption by calcium and phosphorus. Thus calcium and phosphorus atoms were the predominant targets for the absorption events leading to the inactivation of spores in the wavelength range examined.

Wavelength dependence (150-290 nm) of the formation of the cyclobutane dimer and the (6-4) photoproduct of thymine.

The action cross sections for the formation of the cyclobutane dimer and the (6-4) photoproduct of thymine as well as the absorption cross sections of thymine were determined in the wavelength region between 150 and 290 nm. Thymine films sublimed on glass plates were irradiated by monochromatic photons in a vacuum; the induced photoproducts were quantitatively analyzed by high-performance liquid chromatography (HPLC). Under our conditions, two major peaks appeared on the HPLC chromatograms of irradiated samples. The two peaks were identified as being the cis-syn cyclobutane dimer and the (6-4) photoproduct, based on their HPLC retention times, absorption spectra in the effluent, and photochemical reactivity. The fractions of the two photoproducts increased linearly with the fluence at low fluences over the entire wavelength range. Their action cross sections were determined by the slopes of the linear fluence response curve at 10 nm intervals between 150 and 290 nm. The two action spectra showed a similar wavelength dependence and had a maximum at 270 nm as well as two minor peaks at 180 and 220 nm, at which wavelengths the peaks of the absorption spectrum of thymine sublimed on a CaF2 crystal plate appeared. The quantum yields had relatively constant values of around 0.008 for the dimer and 0.013 for the (6-4) photoproduct above 200 nm, decreasing to 0.003 and 0.006, respectively, at 150 nm as the wavelength became shorter.

Repair of ultraviolet light damage in Saccharomyces cerevisiae as studied with double- and single-stranded incoming DNAs.

Purified double- and single-stranded DNAs of the autonomously replicating vector M13RK9-T were irradiated with ultraviolet light (UV) in vitro and introduced into competent whole cells of Saccharomyces cerevisiae. Incoming double-stranded DNA was more sensitive to UV in excision repair-deficient rad2-1 cells than in proficient repair RAD+ cells, while single-stranded DNA exhibited high sensitivity in both host cells. The results indicate that in yeast there is no effective rescue of UV-incoming single-stranded DNA by excision repair or other constitutive dark repair processes.

Iron(II) sulphate (Fricke solution) oxidation yields for 8.9 and 13.6 keV X-rays from synchrotron radiation.

The oxidation yields (G) for 8.86 and 13.55 keV X-rays produced by synchrotron radiation were measured using an iron(II) sulphate (Fricke) solution. Monoenergetic X-rays were produced using a silicon crystal monochromator. The X-rays were absorbed in 0.4 M sulphuric acid-iron(II) sulphate solution and FeIII ion yields were measured and corrected for escape fractions resulting from scattering using Monte Carlo calculations. Doses in the solution were determined using a thin window, parallel plate chamber calibrated against a primary standard free-air chamber at the Electrotechnical Laboratory (Osaka, Japan). Yields (G) of 1.50 +/- 0.06 and 1.43 +/- 0.06 mumol J-1 were obtained for 8.86 and 13.55 keV X-rays respectively.

Inactivation action spectra of Bacillus subtilis spores in extended ultraviolet wavelengths (50-300 nm) obtained with synchrotron radiation.

Five types of Bacillus subtilis spores (UVR, UVS, UVP, RCE, and RCF) differing in repair and/or recombinational capabilities were exposed to monochromatic radiations at 13 wavelengths from 50 to 300 nm in vacuum. An improved biological irradiation system connected to a synchrotron radiation source was used to produce monochromatic UV radiation in this extended wavelength range with sufficient fluence to inactivate bacterial spores. From the survival curves obtained, the action spectra for the inactivation of the spores were depicted. Recombination-deficient RCE (recE) and RCF (recF) spores were more sensitive than the wild-type UVR spores in the entire range of wavelengths. This was considered to mean that DNA was the major target for the inactivation of the spores. Vacuum-UV radiations of 125-175 nm were effective in killing the spores, and distinct peaks of the sensitivity were seen with all types of the spores. Insensitivities at 190 and 100 nm were common to all five types of spores, indicating that these wavelengths were particularly impenetrant and absorbed by the outer layer materials. The vacuum-UV peaks centering at 150 nm were prominent in the spores defective in recombinational repair, while the far-UV peaks at around 235 and 270 nm were prominent in the UVS (uvrA ssp) and UVP (uvrA ssp polA) spores deficient in removal mechanisms of spore photoproducts. Thus, the profiles of the action spectra were explained by three factors; the penetration depth of each radiation in a spore, the efficiency of producing DNA damage that could cause inactivation, and the repair capacity of each type of spore.