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Biomedical subjects

I Witte

Publications and source records attributed to I Witte.

At least 19 recordsLinked to original sources
PubMed2004

Synergistic DNA damage by oxidative stress (induced by H2O2) and nongenotoxic environmental chemicals in human fibroblasts.

Genotoxic combination effects of oxidative stress (induced by H2O2) and eight nongenotoxic environmental chemicals (4-chloroaniline, 2,3,4,6-tetrachlorophenol, lindane, 2,4-dichloroacetic acid (2,4-D), m-xylene, glyphosate, nitrilotriacetic acid and n-hexanol) were determined in human fibroblasts. Genotoxicity was measured quantitatively by the single cell gel electrophoresis assay. The nongenotoxic chemicals were used in non cytotoxic concentrations. H2O2 was used in concentrations producing low (50 microM) and no cytotoxicity (40 microM). All environmental chemicals acted in a synergistic way with H2O2 except DMSO which effectively inhibited H2O(2)-induced DNA damage. The most effective enhancers were 4-chloroaniline, 2,3,4,6-tetrachlorophenol, m-xylene, and n-hexanol. Synergistic effects of hexanol/H2O2 were still evident at a concentration of 0.09 noec (no observed effect concentration). In contrast to synergistic DNA damage in the cell antagonism was found measuring DNA breakage in isolated PM2 DNA. From the results we concluded that synergisms between H2O2 and nongenotoxic chemicals may be a general phenomenon which is not observed on the level of isolated DNA.

Cell Line↗
PubMed2000

Synergistic DNA damaging effects of 4-nitroquinoline-1-oxide and non-effective concentrations of methyl methanesulfonate in human fibroblasts.

DNA damage and DNA repair in human fibroblasts induced by the combination mixture of the genotoxic agents methyl methanesulfonate (MMS) and 4-nitroquinoline-1-oxide (4-NQO) were studied using the comet assay and the unscheduled DNA synthesis (UDS), respectively. Cells were simultaneously treated for 1h with the no observed effect concentration (noec) of MMS and increasing concentrations of 4-NQO or vice versa. Different results were obtained with the two types of mixtures. When the noec of 4-NQO was combined with increasing concentrations of MMS, no combination effects were observed. However, in experiments with increasing concentrations of 4-NQO and the noec of MMS, an increase in DNA damage and repair (and an enhancement of cytotoxicity) was demonstrated. Quantitative analysis of the effects by the isobologram method confirmed synergistic responses in both tests. We are proposing interactive actions between 4-NQO and MMS, whereby 4-NQO facilitates the attack of MMS on the DNA bases.

4-Nitroquinoline-1-oxide↗
PubMed2000

Protection by desferrioxamine and other hydroxamic acids against tetrachlorohydroquinone-induced cyto- and genotoxicity in human fibroblasts.

Tetrachlorohydroquinone (TCHQ) has been identified as a major toxic metabolite of the widely used wood preservative pentachlorophenol and has also been implicated in its genotoxicity. We have recently demonstrated that protection by the trihydroxamate iron chelator desferrioxamine (DFO) on TCHQ-induced single-strand breaks in isolated DNA was not the result of its chelation of iron but rather of its efficient scavenging of the reactive tetrachlorosemiquinone (TCSQ) radical. In this study, we extended our research from isolated DNA to human fibroblasts. We found that DFO provided marked protection against both the cyto- and genotoxicity induced by TCHQ in human fibroblasts when it was incubated simultaneously with TCHQ. Pretreatment of the cells with DFO followed by washing also provided marked protection, although less efficiently compared with the simultaneous treatment. Similar patterns of protection were also observed for three other hydroxamic acids (HAs): aceto-, benzo-, and salicylhydroxamic acid. Dimethyl sulfoxide, an efficient hydroxyl radical scavenger, provided only partial protection even at high concentrations. In vitro studies showed that the HAs tested effectively scavenged the reactive TCSQ radical and enhanced the formation of the less reactive and less toxic 2,5-dichloro-3, 6-dihydroxy-1,4-benzoquinone (chloranilic acid). The results of this study demonstrated that the protection provided by DFO and other HAs against TCHQ-induced cyto- and genotoxicity in human fibroblasts is mainly through scavenging of the observed reactive TCSQ radical and not through prevention of the Fenton reaction by the binding of iron in a redox-inactive form.

Cell Line↗
PubMed1999

Induction of lipid peroxidation in human fibroblasts by the antioxidant propyl gallate in combination with copper(II).

The antioxidant propyl gallate (PG) induced lipid peroxidation in combination with non-toxic Cu(II) concentrations in human fibroblasts. This was measured by the thiobarbituric acid assay (TBA assay) and by detection of accumulating fluorescent products after a 1-h treatment of cells with CuCl2/PG at concentrations higher than 0.125 mM. PG alone led to a significant reduction of thiobarbituric acid-reactive substances (TBARS) demonstrating its antioxidative properties. Time course studies of lipid peroxidation by PG/Cu(II) showed that formation of TBARS was preceded by a lag phase of 60 min. Thereafter, the TBARS value increased rapidly for 1 h and then reached a constant maximum or slightly decreased. The induction of lipid peroxidation by PG/Cu(II) is probably due to the formation of reactive species like reactive oxygen species (ROS), Cu(I) and semiquinone radicals which are able to participate in initiation and propagation of lipid peroxidation. Combination effects of PG/Cu(II) were demonstrated also on inhibition of membrane-bound succinate dehydrogenase. Cytosolic esterases were affected only slightly. The greater susceptibility of membrane-bound enzymes is in accordance with the lipid peroxidation-inducing effects of PG/Cu(II).

Antioxidants↗
PubMed1998

Cytotoxicity and mutagenicity of a 2,4,6-trinitrotoluene (TNT) and hexogen contaminated soil in S. typhimurium and mammalian cells.

The toxicity and mutagenicity of aqueous and organic extracts of soil contaminated with TNT, TNT metabolites and hexogen was determined in mammalian cell lines and in prokaryotic cells. The prokaryotic toxicity was determined via the colony forming ability of Salmonella typhimurium (strains TA 98 and TA 100). The same strains were used to test mutagenicity in the Ames test. The mammalian toxicity was analyzed in human fibroblasts by the inhibition of cell growth and cell viability (MTT assay). The mammalian mutagenicity was tested with the HPRT test in V79 cells (hamster lung). The aqueous soil extract did not reveal toxicity or mutagenicity in any of the tests performed. The DMSO/ethanol extract showed toxicity and mutagenicity in S. typhimurium. Thereby strain TA 98 was more sensitive than strain TA 100. In human fibroblasts cell growth was strongly inhibited, whereas no reduction of cell viability was found in the MTT test. Mutagenicity of the DMSO/ethanol extract of the soil was demonstrated in V79 cells.

Animals↗
PubMed1998

Synergistic DNA damaging effects of malondialdehyde/Cu(II) in PM2 DNA and in human fibroblasts.

Malondialdehyde (MDA) is a product of lipid peroxidation (LPO). In combination with CuCl2 MDA induced single strand breaks in PM2 DNA whereas MDA or CuCl2 alone had no effect. Cu(II) oxidized MDA by a radical mechanism under formation of Cu(I). DNA strand break induction was inhibited by catalase (98%), neocuproine (76%) and DMSO (61%). The synergistic damaging effect of MDA and Cu(II) was also demonstrated in human fibroblasts measured by alkaline elution. The combination MDA/CuCl2 caused extensive DNA breakage while neither MDA nor CuCl2 alone induced DNA damage within the cell. Synergistic cytotoxic effects were observed 18 h after a simultaneous treatment of the cells with MDA and CuCl2 for 1 h.

Administration, Topical↗
PubMed1998

DNA strand break induction and enhanced cytotoxicity of propyl gallate in the presence of copper(II).

The antioxidant propyl gallate (PG) induced single strand breaks in PM2 DNA at concentrations higher than 0.25 microM when it was combined with copper concentrations at 5 microM and above. In combination with 100 microM CuCl2, extensive double strand breakage was also observed. Neither PG alone nor CuCl2 showed any strand breaking properties. DNA strand breakage was inhibited by addition of catalase or the Cu(I) chelator neocuproine, indicating the involvement of H2O2 and a Cu(II)/Cu(I) redox cycle in the DNA damage. DNA damage of PG/Cu(II) was also observed in human fibroblasts. Using the alkaline elution technique concentrations of 0.15-0.5 mM PG induced DNA strand breaks in combination with 2.5 mM CuCl2, while the single substances did not show any effect. At these concentrations cell viability measured by the MTT assay was not reduced by more than 10%; however, cell growth was inhibited by PG in combination with Cu(II). This growth inhibition was apparently due to the DNA damage incurred by PG/Cu(II). The synergistic interaction between PG and Cu(II) is probably caused by a redox reaction between both compounds, whereby reactive species such as ROS are formed, which are responsible for the observed genotoxic and cytotoxic effects. Our results demonstrate that the antioxidative and cytoprotective properties of propyl gallate may change to prooxidative, cytotoxic and genotoxic properties in the presence of Cu(II).

Catalase↗
PubMed1998

Different DNA damaging species as a result of oxidation of n-butyraldehyde and iso-butyraldehyde by Cu(II).

The isomers n- and iso-butyraldehyde (BuA) in combination with Cu(II) induced single and double strand breaks in PM2 DNA, whereas the aldehydes, or Cu(II) alone had only negligible effect. The DNA damage was the result of radical oxidations of the aldehydes under formation of Cu(I). Cu(I) formation was independent of molecular oxygen. Extensive DNA degradation was only observed in the presence of molecular oxygen. Characterization of DNA damage pointed to different ultimate DNA damaging species. While catalase and neocuproine inhibited strand break formation induced by iso-BuA/Cu(II) to a high degree, these inhibitors were less effective in the n-BuA/Cu(II) reaction. On the other hand, sodium azide showed a high strand break inhibition in the n-BuA/Cu(II) reaction, but low inhibition in the iso-BuA/Cu(II) reaction. 2-Deoxyguanosine was hydroxylated in the 8-position by iso-BuA/Cu(II) but little reaction occurred with n-BuA/Cu(II). Chemiluminescence was detected during both BuA/Cu(II) reactions, whereby the intensity of the luminescence signal was 3.5-fold higher for n-BuA/Cu(II) than for iso-BuA/Cu(II). We suppose that the copper(II)-driven oxidation of n- and iso-BuA proceeds via different pathways with different DNA damaging consequences. Whereas the oxidation of iso-BuA mainly results in damage by .OH-radicals, the oxidation of n-BuA may lead to a radical reaction chain whereby excited states are involved and the resulting DNA-damaging species are not .OH-radicals.

8-Hydroxy-2'-Deoxyguanosine↗
PubMed1996

Correlation of the lipophilicity of xenobiotics with their synergistic effects on DNA synthesis in human fibroblasts.

The binary combination effects of DNA synthesis of human fibroblasts were investigated using 2,4-D with 15 xenobiotics of different chemical substance classes. Results were compared with previous investigations on cell growth. Each of the 15 chemicals tested at their no effect concentrations (NOEC's) increased the effects of 2,4-D on DNA synthesis. Thereby, the EC20 value of 2,4-D was reduced by approximately 40% in the combinations. The NOEC's of the xenobiotics used in the combinations varied by a factor of 1,600 and depended strongly on the lipophilicity of the agents combined with 2,4-D. A significant statistical correlation of r = 0.90 was found between the NOEC's of the 15 combined xenobiotics and their lipophilicity. The combination effects on DNA synthesis were similar to those on cell growth. The regression lines of the relationship between the NOEC's and lipophilicity in both assays showed only slight differences in the slopes. This is an additional confirmation of our hypothesis on a facilitated uptake of 2,4-D in the binary combinations.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1996

Suitability of different cytotoxicity assays for screening combination effects of environmental chemicals in human fibroblasts.

Cytotoxicity screening assays measuring survival, growth, colony forming ability, DNA and protein synthesis in human fibroblasts were tested for their suitability to determine combination effects. Thereby, the dose-response curves of a hydrophilic substance A alone and after pretreatment with a membrane damaging substance B were compared. Substances B were applied at concentrations which did not induce toxic effects in the assays (noec). Synergistic combination effects were demonstrated by reduction of the EC20 value of substances A in the combination in comparison to substances A alone. The following substance pairs (substance B/substance A = membrane damaging/hydrophilic) were tested: n-dodecylbenzenesulfonic acid/2,4-dichlorophenoxyacetic acid, 2,4,6-trichlorophenol/2,4-dichlorophenoxyacetic acid, 1,1,2,2-tetrachloroethane/4-chloroaniline, pentachlorophenol/CrCl3. While survival, growth, and DNA synthesis assays were suitable methods for detecting synergistic combination effects, the growth assay was the most sensitive. Here, all four substance pairs showed synergistic combination effects.

Cell Division↗
PubMed1996

DNA single and double strand breaks induced by aliphatic and aromatic aldehydes in combination with copper (II).

The aliphatic n-butyr-and n-valeraldehyde as well as the aromatic benz- and anisaldehyde induced DNA strand breaks in PM2 DNA in the presence of CuCl2. Neither aldehydes nor CuCl2 alone showed DNA breakage properties. The maximum of single strand breaks (SSBs) induced by the combination of CuCl2 and aldehydes was dependent on the CuCl2-concentration. The aliphatic aldehydes induced SSBs and double strand breaks (DSBs) at lower concentrations than aromatic aldehydes when optimal CuCl2 concentration were used. Catalase and neocuproine nearly completely inhibited strand break formation induced by aromatic aldehydes/CuCl2. The prevention of strand breaks induced by aliphatic aldehydes/CuCl2 was less effective. While the inhibition by neocuproine was only 25%, catalase was totally ineffective. In all aldehydes/CuCl2 mixtures the formation of Cu(I) was observed. The results point to different DNA damaging species produced during redox reactions of aromatic and aliphatic aldehydes in combination with CuCl2.

Aldehydes↗
PubMed1995

Correlation of synergistic cytotoxic effects of environmental chemicals in human fibroblasts with their lipophilicity.

The cytotoxic combination effects of 2,4-D with 12 xenobiotics having different lipophilicity were investigated in human fibroblasts at their no effect concentrations (NOEC). Each of the chemicals tested in binary combinations enhanced the toxicity of 2,4-D. These synergistic combination effects were independent of the chemical structure of the test compounds. However, the NOEC's of the xenobiotics used in the combinations varied by a factor of 10,000. For strongly lipophilic compounds the lowest NOEC's were needed to induce synergistic cytotoxicity. A linear regression analysis of the concentrations (NOEC's) of the 12 combined xenobiotics against their lipophilicity revealed a correlation with r = 0.96 for 11 agents. This close correlation may be explained by the membrane damaging properties of lipophilic compounds which enhance the uptake of hydrophilic agents.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1995

Enhanced cyto- and genotoxicity of tetracycline in Wilson disease fibroblasts.

Tetracycline (TC) exerts DNA damaging properties which are accelerated in the presence of copper(II). Thereby, reactive oxygen species are generated. We investigated, if copper-accumulating cells show a higher sensitivity to TC compared to normal cells. Fibroblasts with an increased copper content were derived from patients of two genetic disorders, Wilson disease (WD) and Menkes disease (MD). Cytotoxic and genotoxic effects of TC were investigated in different human fibroblasts. The inhibition of cell growth by TC was measured in two normal fibroblast lines, fibroblast lines of two patients with WD and one patient with MD. While TC inhibited cell growth at similar concentrations in normal fibroblasts and the MD fibroblasts, the WD cells were much more sensitive. Furthermore, an increased inhibition of DNA synthesis and an enhanced induction of unscheduled DNA synthesis (UDS) was found in WD cells after a TC-treatment compared to normal cells.

Cell Division↗
PubMed1994

A comparison of the DNA-damaging, the cytotoxic and genotoxic properties of tetracycline in human fibroblasts in the presence and absence of light.

The reduction of the colony-forming ability, the induction of DNA strand breaks and DNA repair were determined in human fibroblasts after treatment with tetracycline (TC) in the presence and absence of light. In all experiments human fibroblasts were more sensitive to incubations of TC in the light than in the dark. Induction of DNA single-strand breaks and DNA repair were detected in the cells after a 1-h incubation with TC under light but not in the dark. In contrast to these results, TC induced single-strand breaks in isolated PM2 DNA in the dark, however, to a lower extent than in the presence of light. In both cases strand break formation was totally suppressed by adding catalase. The formation of a TC-derived radical by ESR and a decomposition product by UV-vis spectroscopy was observed in the presence and absence of light; their rate of formation in the dark was much smaller than in the light.

Cell Line↗
PubMed1994

Induction and mechanism of DNA single- and double-strand breaks by tetracycline/Cu(II) in the absence of light.

In the absence of light, tetracycline (TC) induced single- and double-strand breaks in PM2 DNA at micromolar concentrations in combination with CuCl2, whereas TC or CuCl2 alone had no effect. Strand break formation was completely suppressed by catalase and the specific Cu(I) scavenger neocuproine. The extent of strand break formation depended on the ratio of Cu(II):TC. At a ratio of > or = 2 most DNA damage was observed. The influence of the kind of Cu(II)/TC complexation on DNA strand break formation is discussed. The DNA damage in PM2 DNA provoked by TC/CuCl2 was indirectly detected also in human fibroblasts by the induction of DNA repair. The results are discussed with regard to human risk from TC/Cu(II).

Bacteriophages↗
PubMed1992

Synergistic effects of Cu(II) and dimethylammonium 2,4-dichlorophenoxyacetate (U46 D fluid) on PM2 DNA and mechanism of DNA damage.

Dimethylammonium 2,4-dichlorophenoxyacetate (2,4-D . DMA) induced strand breaks in PM2 DNA when incubated with CuCl2, whereas 2,4-D . DMA alone or CuCl2 alone did not show any or only a negligible effect. The formation of single strand breaks increased linearly with time and concentration of 2,4-D . DMA. Neocuproine, a specific Cu(I) chelator totally prevented strand break formation. So did catalase (up to 100 mM 2,4-D . DMA), but DMSO had only a small protective effect. 2,4-Dichlorophenol, CO2 and formaldehyde were detected as reaction products of 2,4-D and CuCl2. From these results a redox reaction of Cu(II) and 2,4-D is proposed, which could explain the DNA damaging properties of CuCl2/2,4-D . DMA.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1991

Synergistic effects of U46 D fluid (dimethylammonium salt of 2,4-D) and CuCl2 on cytotoxicity and DNA repair in human fibroblasts.

The cytotoxicity of U46 D Fluid was tested in human fibroblasts after pretreatment with non-toxic or slightly toxic concentrations of CuCl2. While cell survival, colony-forming ability and protein synthesis were not affected by pretreatment with CuCl2, the inhibition of cell growth was enhanced as was inhibition of DNA synthesis. Synergistic effects of CuCl2 and U46 D Fluid were also detected on the induction of DNA repair measured by unscheduled DNA synthesis. While neither U46 D Fluid nor CuCl2 alone induced DNA repair, preincubation with CuCl2 followed by treatment with U46 D Fluid strongly provoked DNA repair.

2,4-Dichlorophenoxyacetic Acid↗