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Biomedical subjects

M V Lafleur

Publications and source records attributed to M V Lafleur.

At least 55 records · Page 3Linked to original sources
PubMed1986

Radiation damage to phi X174 DNA and biological effects.

Dilute aqueous solutions of biologically active DNA can serve as a simplified model system of the cell. As a biological endpoint the survival of the DNA (after transfection to E. coli spheroplasts) is used. Damage in the DNA, irradiated in water with gamma rays, can be ascribed to reactions with primary waterradicals. By introducing additives in such solutions, which will scavenge the primary waterradicals, competition between a scavenger and DNA for such radicals can be studied. Comparison of different additives makes it possible to decide whether a compound behaves like a simple scavenger, radiosensitizer or like a radioprotector. In this context work has been done with the electron-affinic radiosensitizers metronidazole, misonidazole and nifuroxime. We have found that these wellknown cellular sensitizers do not enhance the inactivation of biologically active DNA. They act as simple competitive scavenger for waterradicals. However, if besides a sensitizer a trace of a metalloporphyrin containing compound (e.g. cyt. c) is present during irradiation an enhanced DNA inactivation, which can be interpreted as sensitization, is observed. Without sensitizer metalloporphyrins induce an enhanced protection of DNA. Apart from these effects the consequences of both chemical-(sulphydryl) and enzymatic-(excision; recombination) repair has been studied. It has been found that sulphydryl compounds are able to react with DNA radicals, modifying the radiation damage in such a way that e.g. breaks are prevented. Further in double-stranded DNA a considerable amount of OH and also H radical damage appeared to be reparable by the excision-repair mechanism. However, post-replication repair had only very small or no effect on the amount of damage.

Bacteriophage phi X 174↗
PubMed1986

Effect of radiation-induced reduction of nitroimidazoles on biologically active DNA.

Radiation-chemical reductions have been carried out with several nitroimidazoles. Reduction of these drugs in the presence of single-stranded phi chi 174 DNA causes extensive lethal damage. However, relatively stable (end) products, do not contribute to the damage, although glyoxal is potentially toxic. This demonstrates that a short-lived intermediate in the reduction process is responsible. Further, the quantity of damage in the DNA depends on both dose (reduction)-rate and also the nature of the drug.

Coliphages↗
PubMed1985

Inactivation of phi X174 DNA by the ortho-quinone derivative or its reduction product of the antitumor agent VP 16-213.

Biologically active phi X174 DNA is inactivated by the ortho-quinone derivative of the antitumor agent VP 16-213, but not by VP 16-213 itself, VP 16-213 phenoxy radical or aqueous decomposition product(s) of the ortho-quinone. Reduction of the ortho-quinone by cytochrome P-450 reductase and NADPH results in deactivation of the ortho-quinone towards anti-phi X174 DNA activity. However, compared with the parent compound VP 16-213, reduction of the ortho-quinone results in substantial damage towards DNA.

Bacteriophage phi X 174↗
PubMed1985

Electrolytic reduction of nitroheterocyclic drugs leads to biologically important damage in DNA.

The effect of electrolytic reduction of nitroimidazole drugs on biologically active DNA was studied. The results show that reduction of the drugs in the presence of DNA affects inactivation for both double-stranded (RF) and single-stranded phi X174 DNA. However, stable reduction products did not make a significant contribution to the lethal damage in DNA. This suggests that probably a short-lived intermediate of reduction of nitro-compounds is responsible for damage to DNA.

Bacteriophage phi X 174↗
PubMed1984

Contrasting effects of cytochrome C on the radiosensitivity of single-stranded phi X174 DNA in the presence of misonidazole or phenol under anoxia.

The effect of misonidazole and cytochrome c on biologically active DNA was studied. In a solution of single-stranded phi X174 DNA the presence of both sensitizer and cytochrome c is needed to evoke radiosensitization. However, sensitizer or cytochrome c alone protects the DNA. Cytochrome c offers even an extra protection. The sensitization is dependent on the concentration of misonidazole. This finding opens up the possibility of studying sensitizers in a relatively simple system.

Bacteriophage phi X 174↗
PubMed1984

Repair of damage in double-stranded phi X174 (RF) DNA due to radiation-induced water radicals.

Experiments in which the yields of radiation-induced OH and H radicals were varied, showed that both types of water radicals inactivate phi X174 RF DNA to about the same extent as measured by transfection of the (irradiated) DNA to E. coli wild-type spheroplasts. On the other hand, using spheroplasts prepared from E. coli strains, deficient in one of the proteins involved in excision DNA repair (uvrA- or uvrC-) or in post-replication repair (recA-), clear differences between damage originating from OH or H radical attack were found. Part of the radiation damage due to H radicals appeared to be repairable by an uvrA-gene-dependent repair mechanism, whereas this repair pathway does not play an important role in the case of OH radical damage. The reverse applies to uvrC-gene-dependent repair, which only affects OH radical damage (obtained under anoxic conditions), but has no influence on damage due to H radicals. Irradiation of double-stranded phi X174 (RF) DNA in the presence of oxygen however, yields damage--due to OH radicals only--which appeared not to be sensitive to either uvrC- or uvrA-gene-dependent repair. Furthermore, post-replication repair (recA) has only very little effect on the amount of inactivation by H or OH radicals, when irradiation is carried out under anoxic conditions. We did not find significant inactivation due to hydrated electrons, whether the biological activity was determined by use of wild-type spheroplasts or of strains deficient in excision or post-replication repair proteins.

Bacteriophage phi X 174↗
PubMed1983

Heat-sensitive radiation damage in single-stranded DNA irradiated in a bacterial extract.

The effect of heat treatment (45 degrees C) on gamma-irradiated biologically active single-stranded phi X174 DNA, dissolved in a bacterial extract, was studied. The results show that under these circumstances heat-sensitive damage is found, which is absent after irradiation in pure buffer. The damage is non-lethal and probably becomes an apurinic/apyrimidinic site, which is lethal, upon the post-irradiation heat treatment. Prolonged heating converts it into a break. The amount of the initial damage depends on the conditions of irradiation.

Bacteriophage phi X 174↗
PubMed1982

Influence of anoxic sensitizers on the radiation damage in biologically active DNA in aqueous solution.

The competition between biologically active single-stranded phiX174 DNA and the anoxic radiosensitizers metronidazole, misonidazole, paranitroacetophenone or nifuroxime for OH radicals is studied. The results are compared with experiments in which the protection of the DNA by t-butanol is determined. Also the effects of the sensitizers on the chemical nature of the damage (immediate and potential break, immediate and potential base damage) is studied. It is found that in diluted aqueous solutions of DNA these radiosensitizers do not sensitize with respect to the biological inactivation. The only effect observed is a shift from potential to immediate breaks with misonidazole and also nifuroxime.

Acetophenones↗
PubMed1981

Some characteristics of apurinic sites in single- and double-stranded biologically active phi X174 DNA.

With biologically active DNA of the bacteriophage phi X174, both single and double-stranded, some physico-chemical and biological parameters of the depurination reaction are studied. It is shown that in single-stranded DNA each apurinic site is lethal, while in double-stranded RFI-DNA only about 5% of these sites are lethal. Furthermore it is concluded that the apurinic sites are formed at different rates in single- and double-stranded DNA and also the conversion into breaks of the apurinic sites is different for both forms of DNA.

Apurinic Acid↗
PubMed1981

Alkali-labile sites in biologically active DNA: comparison of radiation induced potential breaks and apurinic sites.

Gamma-ray induced alkali-labile sites, which are converted into breaks by alkali or heat, are compared with apurinic sites in both single-stranded and phi X174RF-DNA. It is shown that the kinetics of the conversion into breaks are completely different for both sites. The conclusion is: the gamma-ray induced alkali-labile sites are not identical with apurinic or/and apyrimidinic sites.

Alkalies↗
PubMed1980

Effects of sulphydryl compounds on the radiation damage in biologically active DNA.

The effect of sulphydryl compounds on the induction of alkali-labile sites and on the contribution of such sites to the inactivation of single-stranded phi X174 DNA was studied. Cysteamine is capable of reacting with DNA radicals, thereby modifying the radiation damage in such a way that the induction of immediate and latent breaks is reduced. This depends on the pH of the solution. With cysteine only, a pH dependent protection, against lethal alkali-labile potential breaks could be observed. The damage other than breaks is not influenced by the presence of sulphydryl compounds.

Alkalies↗
PubMed1979

Alkali-labile sites and post-irradiation effects in gamma-irradiated biologically active double-stranded DNA in aqueous solution.

Gamma-irradiation of double-stranded RF-DNA of bacteriophage phi X174 in aqueous solution in the presence of oxygen produces at least one type of alkali-labile site. It is lethal and gives rise to breaks by alkali and is identical with the damage which becomes manifest by post-irradiation heat treatment. The effect of alkali is dependent on temperature. Furthermore, the excision repair system is not involved in eliminating lethal nucleotide damage in RFI-DNA.

Alkalies↗
PubMed1978

Alkali-labile sites and post-irradiation effects in single-stranded DNA induced by H radicals.

Single-stranded phiX174 DNA in aqueous solutions has been irradiated in the absence of oxygen, under conditions in which only H radicals react with the DNA. It was shown that H radical reactions result in breaks, which contribute approximately 10 per cent inactivation. Further, two types of alkali-labile sites are formed. One is lethal and gives rise to single-strand breaks by alkali and is most probably identical with post-irradiation heat damage and contributes about 33 per cent to the inactivation mentioned above. The other consists of non-lethal damage, partly dihydropyrimidine derivatives, and is converted to lethal damage by alkali. This follows from experiments in which the DNA was treated with osmium-tetroxide, which oxidizes thymine to 5,6-dihydroxy-dihydrothymine. Treatment with alkali of this DNA gives the same temperature dependence as found for the non-lethal alkali-labile sites in irradiated DNA. A similar temperature dependence is found for dihydrothymine and irradiated pyrimidines with alkali.

Alkalies↗