PubMed Health⌕ Search

Biomedical subjects

M Näslund

Publications and source records attributed to M Näslund.

At least 19 recordsLinked to original sources
PubMed1998

MNNG-induced [corrected] RecBCD dependent DNA degradation in recA13 mutant cells is not the basis of their hypersensitivity to this agent.

We have examined the hypersensitivity of Escherichia coli recA13 mutant cells to killing by N-methyl-N'-nitro-N-nitro-soguanidine (MNNG) and have shown out that despite MNNG-induced adaptation they remained vastly more sensitive to the cytotoxic effect of this agent than wild type cells. Because this might have been a consequence of a different extent of induction of the adaptive response in the recA13 background, we have measured O6-alkylguanine-DNA alkyltransferase (ATase) activity in extracts of adapted and non-adapted recA13 mutant and wild type cells. Adaptation increased ATase levels by 28- and 34-fold in wild type and recA13 mutant cells, respectively. Thus, the adaptive response was no less inducible in recA13 mutant cells than in wild type cells. This indicates that the extreme sensitivity of recA13 cells to MNNG is not caused by an inability to repair the principal toxic lesions induced in DNA. Low doses of MNNG caused substantial degradation of cellular DNA in recA13 mutant cells but not in the wild type cells. This DNA degradation is shown to be the RecBCD-enzyme dependent. Since recA13 recB21 double mutants were even more sensitive to MNNG than recA single mutants, DNA degradation appears not to be the cause of the MNNG-hypersensitivity in recA13 cells.

Adaptation, Biological↗
PubMed1998

Rejoining of DNA strand breaks induced by propylene oxide and epichlorohydrin in human diploid fibroblasts.

The repair kinetics of DNA single- and double-strand breaks (SSBs, DSBs) induced with two carcinogenic epoxides, propylene oxide (PO) and epichlorohydrin (ECH), was studied in human diploid fibroblasts. The methods used were: alkaline DNA unwinding (ADU), the comet assay, and pulsed field gel electrophoresis (PFGE). About 70% of SSBs, measured by ADU, were rejoined after the treatment with 5 mMh and 10 mMh of PO within 20 hr, and the half-life was estimated to be approximately 15 hr. On the other hand, effective rejoining of SSBs after ECH treatment was observed only at a dose of 1 mMh (a half-life of approximately 15 hr), whereas after 2 mMh treatment, only 26% of SSBs could be rejoined within 20 hr. Furthermore, the use of the comet assay demonstrated that DNA strand breaks were effectively rejoined after PO and ECH treatment at doses of 5-10 mMh and 0.5-1 mMh, respectively. About 76% and 83% of DSBs induced by 5 and 10 mMh of PO, respectively, were rejoined within 4 hr after the treatment (a half-life of approximately 2.5 hr), with little further repair thereafter. At lower dose of ECH (1 mMh) a half-life for DSBs rejoining was estimated to be approximately 2 hr; however, only 29% of DSBs were rejoined within 2 hr at the higher dose of 2 mMh. After 18 hr, the rejoining following treatment with a lower dose was negligible. At a higher dose, a rapid accumulation of DSBs was observed, probably as the result of cell death and DNA degradation. The results demonstrate the capability of human diploid fibroblasts to repair DNA SSBs and DSBs at low-to-moderate doses of the epoxides. A weak capacity to rejoin DNA strand breaks induced by higher doses of ECH may be a consequence of its higher DNA alkylation activity and approximately 10 times higher toxicity compared to PO.

Carcinogens↗
PubMed1997

Propylene oxide and epichlorohydrin induce DNA strand breaks in human diploid fibroblasts.

The induction of DNA strand breaks in human diploid fibroblasts (VH-10) was demonstrated after in vitro exposure with two carcinogenic epoxides, propylene oxide (PO) and epichlorohydrin (ECH). Alkaline DNA unwinding (ADU), pulsed field gel electropharosis (PFGE), and the comet assay were used to measure DNA single. (SSBs) and double-strand breaks (DSBs). A dose-dependent increase of DNA strand breaks, measured by ADU, was observed in the dose range 2.5-20 mMh of PO and 0.25-2 mMh of ECH. The dose-response of ECH was about five times higher compared with that of PO (211 vs. 41 SSBs. 100 Mbp-1.mMh-1). The induction rates of DSBs, measured by PFGE, were found to be 18 times higher for ECH compared to PO (4.8 and 0.27 DSBs.100 Mbp-1.mMh-1 for ECH and PO, respectively). Using these two methods, the SSBs/ DSBs ratio was estimated to be 148 for PO and 44 for ECH. The data obtained by the comet assay also demonstrated a dose-dependent ability of PO and ECH to induce DNA damage. It was found that ECH was about six times more effective as an inducer of DNA strand breaks compared to PO (200 and 32x100 Mbp-1.mMh-1 for ECH and PO, respectively). The SSBs/DSBs ratios calculated using comet assay and PFGE data were 125 for ECH and 41 for PO. In addition, ECH is about 10 times more toxic than PO with respect to survival. These properties of ECH can at least in part be explained by its higher chemical reactivity connected with a higher rate of DNA alkylation.

Carcinogens↗
PubMed1995

Expression of Escherichia coli recA and ada genes in Saccharomyces cerevisiae using a vector with geneticin resistance.

Construction of E. coli-yeast shuttle plasmids containing the neo selection gene is described. The protein-coding regions of the E. coli ada or recA genes under the control of the ADH1 promoter and terminator were ligated into the SphI unique site of pNF2 to produce pMSada and pMSrecA, respectively. The plasmids were used for transformation of the haploid and diploid pso4-1 strains of S. cerevisiae and their corresponding wild types. Transformants were obtained by selection for geneticin (G418) resistance. Crude protein samples were extracted from the individual transformants. Both the RecA and Ada proteins were present in all strains containing the recA and ada genes on plasmids, respectively. Thus the geneticin selection system was successfully used for the preparation of model yeast strains.

Bacterial Proteins↗
PubMed1992

Inhibition of recA induction by the radioprotector 2-mercaptoethylamine.

Our earlier finding that the radioprotective action of 2-mercaptoethylamine (MEA) is counteracted by ascorbate suggests a biochemical mechanism of action, which is supported by observations that MEA is not radioprotective in Rec- E. coli strains. In this study we show that MEA inhibits the induction of the recA gene by UV- or gamma-irradiation or by nalidixic acid in Escherichia coli strain GE94, which contains a recA-lacZ fusion. This effect, which may be counteracted by cysteine, indicates that in general MEA inhibits the induction of SOS functions.

Cysteamine↗
PubMed1990

Studies of the rad-equivalence of ethylene oxide in the presence and absence of 12-O-tetradecanoylphorbol-13-acetate (TPA) in C3H/10T1/2 cells.

Cell transformation in vitro of C3H/10T1/2 cells, using gamma-radiation and ethylene oxide (EtO), in both the absence and presence of the cancer promoter, 12-O-tetradecanoylphorbol-13-acetate (TPA), was studied. TPA promotes transformation of C3H/10T1/2 cells to the same extent. In the dose ranges studied the average enhancement of the transformation frequency was 2.4 and 2.5 for EtO and gamma-radiation, respectively. The rad-equivalence of EtO in the presence of TPA was calculated to be 75 +/- 52 rad/mMh (95% confidence interval) which is consistent with the value 78 +/- 14 rad/mMh (95% confidence interval) obtained without TPA treatment.

Animals↗
PubMed1989

Modifying action of gamma-radiation in mutagenesis of E. coli WU36-10 induced by ethylene oxide, ethyl methanesulfonate and methyl methanesulfonate.

The influence of gamma-radiation pre-exposure on ethylene oxide, ethyl methanesulfonate and methyl methanesulfonate mutagenesis in Escherichia coli WU36-10 was studied. Pretreatment with gamma-radiation resulted, in the case of subsequent treatment with ethylene oxide and ethyl methanesulfonate, in a decrease of the frequency of leu+ revertants, and in the case of subsequent treatment with methyl methanesulfonate, in an increase of this mutation frequency.

DNA Repair↗
PubMed1989

Comparative studies of in vitro transformation by ethylene oxide and gamma-radiation of C3H/10T1/2 cells.

A dose-dependent transforming ability of the direct-acting alkylating agent, ethylene oxide (EtO), was demonstrated in C3H/10T1/2 mouse embryo fibroblasts. Morphologically transformed colonies were observed 5-6 weeks after the treatment with EtO. The transforming effectiveness of EtO was compared with that of gamma-radiation, and the rad-equivalence of EtO was calculated to be approximately 90 rad/mMh. This value is consistent with earlier estimates based on induced mutation in various biological systems and thus indicates the applicability of the C3H/10T1/2 cell system in the risk estimation of human exposure to genotoxic compounds.

Animals↗
PubMed1988

Methylations in human hemoglobin.

Levels of N-Methylvaline (MeVal) and N tau-methylhistidine (MeHis) were measured in male smokers and non-smokers in a program aimed at mapping background alkylations of hemoglobin (Hb) as potential indicators of doses of exogenous and endogenous genotoxic agents. MeVal was also determined in Hb from rats, Syrian golden hamsters, mice and chickens. MeVal was found to occur at levels around 0.5 nmole/g Hb, with relatively little variation between individuals and species. MeVal was not significantly affected by smoking. This result contrasts with elevated levels of N-hydroxyethylvaline (HOEtVal) measured in the same persons (Törnqvist et al., 1986b). Levels of S-methylcysteine (MeCys) (Bailey et al., 1981) and MeHis were much higher than those of MeVal. The high levels of MeCys and MeHis may be due partly to misincorporation during protein synthesis and to artifacts. S-Adenosylmethionine and formaldehyde are possible endogenous sources of MeVal. One individual (smoker) out of 21 selected for measurement of MeVal was an outlier, with raised levels of both MeVal and HOEtVal, as would be expected in case of a defective detoxification system.

Animals↗
PubMed1986

Mutagenic and comutagenic action of 5'-deoxy-5'-(methylthio)adenosine.

5'-Deoxy-5'-(methylthio)adenosine (MTA) alone and in combination with methionine was found to increase frequencies of background HGPRT mutation and SCE in V79 Chinese hamster cells. The same agents exert a comutagenic action on these effects as well as upon mutation induction in Escherichia coli stain AB1157 (but not in its non-adaptable derivative ada-6) following treatment with N-methyl-N-nitrosourea (MNU). MTA plus methionine also enhanced the lethal action of MNU on hamster cells. The effects observed may tentatively be ascribed to hypomethylation due to inhibition of DNA methylase by MTA.

Adenosine↗
PubMed1986

Enhancement by cysteamine of N-methyl-N-nitrosourea mutagenesis in E. coli.

Cysteamine (MEA) is comutagenic to methylnitrosourea (MNU) in E. coli AB 1157 but not in the nonadaptable mutant derivative ada-6 of that strain. The comutagenic action of MEA was eliminated by cysteine at low concentrations, which also lowered mutation frequencies in AB1157 but not in ada-6. In model experiments it was shown that cysteine counteracted the inhibition by MEA of beta-galactosidase induction in both bacterium strains. The comutagenic action of MEA is interpreted as being due to an inhibition of induction of methyltransferase during treatment with MNU.

Bacterial Proteins↗
PubMed1983

Lack of additive effect in mutagenesis of E. coli by UV-light and ethylene oxide.

The interaction of UV-irradiation and ethylene oxide (EtO) on forward mutation frequencies in the lacI gene of E. coli strains 3835 and 3951 and on the frequency of leu+ revertants in E. coli WU36-10-89 were studied. Pre-exposure to low doses of UV-light with the following treatment by low and intermediate doses of EtO showed lack of additive effect in the mutagenic response in all strains studied. The number of mutants actually obtained in the respective experiments was much lower than the additive model predicted.

DNA Repair↗
PubMed1978

Suppression of induced beta-galactosidase synthesis by cysteamine and its reversion by gamma-irradiation in the presence of ascorbate.

The induced synthesis of beta-galactosidase in E. coli was found to be inhibited by cysteamine. This inhibitory effect of the SH compound was antagonized by the addition of ascorbate followed by gamma-irradiation with relatively low doses. The cAMP level which, it has been suggested, plays a role in the radioprotective action of cysteamine, is stabilized by ascorbate against changes induced by irradiation.

Ascorbic Acid↗
PubMed1976

Role of peroxide in the radioprotective action of thiols in E. coli.

The radioprotective action of cysteamine (MEA) and cysteine in E. coli is due partly to autoxidatively generated hydrogen peroxide (H2O2). This effect, which predominates at low concentrations of the thiols (1-2 mM in neutral solution), is regularly correlated with a metabolic block, measured as inhibition of RNA synthesis. In experiments with E. coli 15 (autotroph) under exponential growth in complete medium, the role of H2O2 was demonstrated by (a) a decreased radioprotective action if catalase was present in the medium; (b) a radioprotective action of H2O2 added to the medium; (c) a decreased protective action in the absence of catalytically active copper; and (d) oxygen being required for the radioprotective action to develop. At higher concentrations of the thiols, their radioprotective action, and the accompanying metabolic block, are less dependent on H2O2 generation and presumably due to a different mechanism. The radioprotective action of H2O2 is possibly related to the radioprotective action in mammals of catalase inhibitors.

Catalase↗