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F Oesch

Publications and source records attributed to F Oesch.

At least 109 records · Page 6Linked to original sources
PubMed1995

Cell-contact mediated modulation of the sialylation of contactinhibin.

Contactinhibin was found to be involved in contact-dependent inhibition of growth. The growth inhibitory activity of contactinhibin is mediated by N-linked oligosaccharides with desialylated beta-glycosidically linked, terminal galactose residues. Here we show that in sparse human fibroblasts contactinhibin was expressed in a biologically inactive, highly sialylated form both on the plasma membrane and intracellularly, while in confluent cells plasma membrane localized contactinhibin was present in a biologically active, low sialylated form. Plasma membranes were shown to contain a glycoprotein sialidase which is suggested to be engaged in the activation of contactinhibin in a cell contact-dependent manner.

Antibodies↗
PubMed1995

Non-competitive inhibition of clomipramine N-demethylation by fluvoxamine.

The selective serotonin reuptake inhibitor fluvoxamine interferes with the metabolism of tricyclic antidepressants. The present investigation was set out to characterize these interactions in vitro using rat liver microsomes and in vivo by analysing levels of clomipramine and metabolites in sera of depressed patients treated concomitantly with fluvoxamine and clomipramine. Clomipramine was N-demethylated and hydroxylated in vitro by microsomes to N-desmethyl-clomipramine, 8-hydroxyclomipramine, and 10-hydroxyclomipramine. Kinetic analyses revealed Km values of 6.2 microM for N-demethylation and 1.2 microM for 8-hydroxylation. Fluvoxamine was a non-competitive inhibitor for N-demethylation with mean Ki value of 6 microM. In the sera of patients treated with daily doses of 150 mg clomipramine and varying doses of fluvoxamine, decrease in the formation of N-desmethylclomipramine and 8-hydroxyclomipramine were found in comparison to those in sera of patients receiving clomipramine as monotherapy. Taken together, the data give evidence that fluvoxamine is a potent non-competitive inhibitor of N-demethylation and to a minor extent of 8-hydroxylation of clomipramine. Because of the species differences in the metabolism of xenobiotics between rodents and humans, conclusions from animal studies on the clinical situation must be drawn cautiously. Nevertheless, the in vitro approach was helpful to understand drug interactions between clomipramine and fluvoxamine in psychiatric patients.

Animals↗
PubMed1995

The gap junctional intercellular communication is no prerequisite for the stabilization of xenobiotic metabolizing enzyme activities in primary rat liver parenchymal cells in vitro.

In primary monocultures of adult rat liver parenchymal cells (PC), the activities of the xenobiotic metabolizing enzymes microsomal epoxide hydrolase (mEHb), soluble epoxide hydrolase (sEH), glutathione S-transferases (GST), and phenolsulfotransferase (ST) were reduced after 7 d to values below 33% of the initial activities. Furthermore, the gap junctional intercellular communication (GJIC), measured after microinjection by dye transfer, decreased from 90% on Day 1 to undetectable values after 5 d in monoculture. Co-culture of PC with nonparenchymal rat liver epithelial cells (NEC) increased (98% on Day 1) and stabilized (82% on Day 7) the homotypic GJIC of PC. Additionally, most of the measured xenobiotic metabolizing enzyme activities were well stabilized over 1 wk in co-culture. Because GJIC is one of several mechanisms playing an important role in cell differentiation, the importance of GJIC for the stabilization of xenobiotic metabolizing enzymes in PC was investigated. PC in monoculture were, therefore, treated with 2% dimethyl sulfoxide (DMSO), a differentiation promoting factor, and 1,1,1-trichloro-2,2,-bis (p-chlorophenyl) ethane (DDT) (10 micrograms/ml), a liver tumor promotor and inhibitor of GJIC, was given to co-cultures of PC with NEC. DMSO significantly stabilized (68% on Day 7), while DDT significantly inhibited (8% on Day 7) homotypic GJIC of PC in the respective culture systems. In contrast, the activities of mEHb, sEH, GST, and ST were not affected in the presence of DMSO or DDT. These results lead to the assumption that the differentiation parameters measured in this study (i.e., homotypic GJIC and the activities of xenobiotic metabolizing enzymes) are independently regulated in adult rat liver PC.

Animals↗
PubMed1995

Viability, attachment efficiency, and xenobiotic metabolizing enzyme activities are well maintained in EDTA isolated rat liver parenchymal cells after hypothermic preservation for up to 3 days in University of Wisconsin solution.

Rat liver parenchymal cells were isolated by EDTA perfusion and were subsequently purified by Percoll centrifugation. The freshly isolated liver cells had a mean viability of 95% as judged by trypan blue exclusion. Isolated liver parenchymal cells were then stored at 0 degrees C for up to 1 wk in University of Wisconsin solution (UW). During this hypothermic preservation, the viability was only slightly reduced to 92% after 1 d and to 85% after 3 d at 0 degrees C. Thereafter, the viability decreased rapidly. After cold storage for up to 3 d, it was possible to use the parenchymal liver cells either in short-term suspension or in cell culture. The attachment efficiency in cell culture was the same for freshly isolated liver cells (84%) and after 2 d cold preservation (81%). The cytochrome P450 content and the enzyme activities of soluble epoxide hydrolase, UDP-glucuronosyl transferase, phenol sulfotransferase, and glutathione S-transferase were not significantly different between freshly isolated cells and cells after 3 d of hypothermic preservation. Furthermore, freshly isolated and intact liver cells stored for 3 d were used in the cell-mediated Salmonella mutagenicity test as a metabolizing system. Both fresh and stored liver parenchymal cells metabolized benzo(a)pyrene,2-aminoanthracene, and cyclophosphamide to their ultimate mutagens.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenosine↗
PubMed1995

Cryopreserved and hypothermically stored rat liver parenchymal cells as metabolizing system in the Salmonella mutagenicity assay.

Freshly isolated and preserved rat liver parenchymal cells were used as metabolizing system in the Salmonella mutagenicity assay. The liver cells were isolated with EDTA perfusion without the addition of collagenase and had a viability of 96% as judged by trypan blue exclusion. When freshly isolated liver parenchymal were cryopreserved with a computer controlled freezing protocol and stored at -196 degrees C they had a mean viability of 89% after thawing. Furthermore, freshly isolated cells were stored at 0 degree C in University of Wisconsin organ transplantation solution. After 1 day of hypothermic storage they had a viability of 95%. Four different indirect mutagens, 2-aminoanthracene, benzo[a]pyrene, 7,12-dimetylbenz[a]anthracene and cyclophosphamide, were used with the liver cells as metabolizing system in the preincubation assay with Salmonella typhimurium TA100. After cryopreservation, liver parenchymal cells were able to activate all tested indirect mutagens to ultimate mutagens. However, the induction of revertants was lower with three of the four tested compounds. Only 2-aminoanthracene was activated to the same extent by freshly isolated and cryopreserved liver cells. 7-Hydroxymethyl-12-methylbenz[a]anthracene, which is activated to its ultimate mutagen by sulfotransferase, also induced a reduced mutagenic effect with cryopreserved liver cells in comparison to freshly isolated liver parenchymal cells. This indicates that phase I and phase II enzyme activities are effected by cryopreservation. However, identical mutation frequencies were obtained when freshly isolated liver parenchymal cells or 1 day hypothermically preserved liver parenchymal cells were used in the cell-mediated Salmonella mutagenicity test. The use of hypothermic short-time storage of liver parenchymal cells could help to make the liver cell-mediated genotoxicity test simpler and thereby more attractive.

Adenosine↗
PubMed1995

DNA damage in nurses handling antineoplastic agents.

In 91 nurses from several divisions of four hospitals in Germany the genotoxic effect caused by the occupational exposure presumably due to mixing of antineoplastic agents was investigated. The amount of DNA single strand breaks and alkali labile sites in the peripheral mononuclear blood cells of the nurses was measured using the alkaline elution method. In ten nurses handling antineoplastic agents not using recommended safety precautions such as safety hoods, gloves or surgical masks a 50% higher level of DNA strand breaks and alkali-labile sites (p < 0.005; U-test) was detected compared to 54 controls. After applying recommended safety precautions a statistically significant decrease (p < 0.01) in the level of DNA strand breaks to the level of controls was observed. In other nurses handling antineoplastic agents by using adequate safety equipment no significantly different amount of DNA strand breaks compared to that of controls was detected. No significant correlation between the level of DNA strand breaks and the weekly contact frequency, the life-time exposure to antineoplastic agents, or the time elapsed since the last handling of the drugs was found in this study.

Adult↗
PubMed1995

DNA damage in mononuclear blood cells of metal workers exposed to N-nitrosodiethanolamine in synthetic cutting fluids.

In 28 smoking and 37 non-smoking male metal workers from 7 small to medium sized plants the genotoxic effect caused by the occupational exposure measured as DNA single strand breaks in the peripheral mononuclear blood cells was investigated. Metal workers using synthetic cutting fluids are possibly exposed to the carcinogenic N-nitrosodiethanolamine (NDELA). In this investigation NDELA was detected in the air of the working halls in a concentration up to 1000 ng/m3 and in the cutting fluids in a concentration up to 135 mg/l (mean values per plant). Workers staying in rooms with a mean concentration of NDELA in the air of about 1 microgram/m3 revealed two times more DNA strand breaks than workers staying in an environment with less than 50 ng/m3 of NDELA (p < 0.01). Non-smoking workers with more than 4.5 h contact to cutting fluids per day showed an 1.5 times higher mean level of DNA strand breaks than their nonsmoking colleagues having had less than 4.5 h contact to cutting fluids (p < 0.02). Also workers having had work place-related complaints showed a statistically significantly higher level of DNA strand breaks compared to workers with no or no work place-related complaints. No significant correlation was obtained between the extent of DNA damage and the estimated extent of skin contact or the concentration of NDELA found in the cutting fluids. Therefore, for workers in this investigation NDELA incorporated by inhalation is probably more relevant for genotoxic damage than NDELA resorbed by skin. An increased level of DNA damage was found in metal workers depending on the concentration of NDELA in the air of the work places. However, without further investigations it cannot be excluded that also other concomitant agents in the environment were responsible for the observed genotoxic effect.

Adult↗
PubMed1995

Gene evolution of epoxide hydrolases and recommended nomenclature.

We have analyzed amino acid sequence relationships among soluble and microsomal epoxide hydrolases, haloacid dehalogenases, and a haloalkane dehalogenase. The amino-terminal residues (1-229) of mammalian soluble epoxide hydrolase are homologous to a haloacid dehalogenase. The carboxy-terminal residues (230-554) of mammalian soluble epoxide hydrolase are homologous to haloalkane dehalogenase, to plant soluble epoxide hydrolase, and to microsomal epoxide hydrolase. The shared identity between the haloacid and haloalkane dehalogenases does not indicate relatedness between these two types of dehalogenases. The amino-terminal and carboxy-terminal homologies of mammalian soluble epoxide hydrolase to the respective dehalogenases suggests that this epoxide hydrolase, but not the soluble epoxide hydrolase of plant or the microsomal epoxide hydrolase, derives from a gene fusion. The homology of microsomal to soluble epoxide hydrolase suggests they derive from a gene duplication, probably of an ancestral bacterial (epoxide) hydrolase gene. Based on homology to haloalkane dehalogenase, the catalytic residues for the soluble and microsomal epoxide hydrolases are predicted. A nomenclature system based on divergent molecular evolution is proposed for these epoxide hydrolases.

Amino Acid Sequence↗
PubMed1995

Malignantly transformed non-parenchymal liver epithelial cells and transformed oval cells suppress the homotypical gap junctional intercellular communication of co-cultured rat liver parenchymal cells.

Isolated rat liver parenchymal cells (PC) were co-cultured with a non-parenchymal rat liver epithelial cell line (NEC) or with an oval cell line. The homotypical gap junctional intercellular communication (GJIC) between the liver PC was measured after microinjection of Lucifer Yellow by dye transfer. The rat liver PC were dye coupled between 87% and 100% for at least 1 week in both co-cultures, in contrast to PC In monoculture between which no dye coupling was left after 1 week. When liver PC were co-cultured with a transformed and tumorigenic NEC or with a transformed and tumorigenic oval cell line the homotypical GJIC between the liver PC was drastically decreased with culture time, and the PC were then compressed and displaced by the expansive growth of the transformed cell lines. The disturbance of the GJIC between normal cells by adjacent tumorigenic cells might be a new mechanism to explain the expansive growth of tumors.

Animals↗
PubMed1995

Malignant transformation of the liver tumour precursor cell line OC/CDE 22 by the four stereoisomeric fjord region 3,4-dihydrodiol 1,2-epoxides of benzo[c]phenanthrene.

In previous work we established the rat liver oval cell line OC/CDE 22 in order to study in vitro mechanisms of liver cell transformation. We have now exposed OC/CDE 22 cells to each of the four optically active fjord region dihydrodiol epoxides of benzo[c]phenanthrene to investigate their capacity for malignant transformation of liver cells. All four configurational isomers, which are among the most potent carcinogenic metabolites of polycyclic aromatic hydrocarbons tested in murine tumour models, malignantly transform OC/CDE 22 cells at a 2 microM dose level, resulting in a similar colony-forming efficiency in soft agar. Inoculation of the transformed cells into newborn syngeneic rats produced an extremely high incidence of carcinomas with a short latency period. The induced carcinomas displayed cholangiocellular, adenoid and solid growing structures. Neither cell growth in soft agar nor induction of tumor formation in newborn rats were achieved if confluent OC/CDE 22 cell cultures were exposed to each of the four stereoisomers and left in the confluent state for 4 weeks. In contrast, if confluent cells were exposed to the four stereoisomers, immediately split and then subcultured as usual, full transformation was accomplished. Our results indicate that the fjord region dihydrodiol epoxides of benzo[c]phenanthrene are highly efficient transforming agents for rat liver cells and that proliferation plays a pivotal role in the liver cell transformation process induced by polycylic aromatic hydrocarbons.

Animals↗
PubMed1995

Detection of primary DNA damage: applicability to biomonitoring of genotoxic occupational exposure and in clinical therapy.

The biological effect of putative genotoxic chemicals in the work place environment was monitored in peripheral mononuclear blood cells of exposed workers. DNA strand breaks, alkali-labile sites of DNA and DNA cross-links were measured using the alkaline filter elution method. A dose dependent increase in DNA damage was found in sterilization workers exposed to ethylene oxide and metal workers with exposure towards N-nitrosodiethanolamine. Two subpopulations with different response to the external exposure were found in nonsmoking sterilization workers. Nurses handling antineo-plastic agents without adequate safety provisions showed a statistically significantly higher rate of DNA strand breaks compared to other nurses handling cytostatic drugs with recommended safety equipment and also compared to not exposed controls. Also in several other occupational groups such as fire fighters possibly exposed to several genotoxic chemicals after an accident in a chemical plant, roofers and petrol pump attendants a significantly higher amount of DNA damage was found compared to controls. No statistically significant differences in the amount of DNA strand breaks were found in cabinet makers and car mechanics compared to controls. In peripheral mononuclear blood cells of ovarial carcinoma patients as well as of patients with Morbus Hodgkin an increased DNA strand break rate was found after application of cytostatic drugs. The individual patients showed a very different response after drug intake. The increase in DNA damage after drug application is possibly related to the success of the chemotherapy.

Animals↗
PubMed1995

Cell systems for use in studies on the relationship between foreign compound metabolism and toxicity.

Since the metabolism of most foreign compounds is predominantly controlled by hepatic in metabolism, isolated hepatocytes in most cases quite well predict the pattern of the overall metabolism of a given compound. Methods have been developed for cryopreserving isolated hepatocytes from man and other species with satisfactory maintenance of foreign compound metabolizing enzyme activities. The installation of a bank of cryopreserved hepatocytes from different species is possible and may be used for rational species extrapolation. It is necessary for some toxicological investigations to have hepatocytes which retain their differentiated status in culture for a sufficient time period. This might be achieved by co-culturing hepatocytes with diverse cell lines. However, from one cell line to the other differences in the pattern of stabilization of individual hepatocyte functions are found. In addition, questions on metabolic action of individual isoenzymes can also be addressed by the use of genetically engineered cell lines. All the in vitro systems mentioned, especially those which contain differentiated human cells or human isoenzymes are helpful in the rational species extrapolation of toxic effects from animal to man.

Animals↗
PubMed1995

Induction of rat liver microsomal epoxide hydrolase by its endogenous substrate 16 alpha, 17 alpha-epoxyestra-1,3,5-trien-3-ol.

1. The influence of the endogenous steroid epoxides 16 alpha, 17 alpha-epoxyestra-1,3,5(10)-trien-3-ol (estroxide) and 16 alpha, 17 alpha-expoxiandrost-4-en-3-one (androstene oxide) and their metabolic precursors estra-1,3,5(10), 16-tetraen-3-ol (estratetraenol) and androsta-4, 16-dien-3-one (androstadienone) on the specific activities of hepatic microsomal and soluble epoxide hydrolase, glutathione S-transferase, dihydrodiol dehydrogenase, and 7-ethoxycoumarin deethylase was investigated in the male Sprague-Dawley rat. 2. Both estroxide and estratetraenol induced microsomal epoxide hydrolase activity towards styrene oxide and estroxide itself 2-2.5-fold and glutathione conjugation of 1-chloro-2,4-dinitrobenzene (CDNB) 1.6-fold after intraperitoneal administration of a high dose of compound (300 mg per kg of body weight). 3. In addition, estroxide decreased 7-ethoxycoumarin deethylation down to 20% of the activity observed in the untreated rat, whereas estratetraenol enhanced the activity of soluble epoxide hydrolase towards trans-stilbene oxide by a factor of 1.7. 4. In contrast, neither androstene oxide nor androstadienone showed a significant influence on any of the parameters under investigation. Dihydrodiol dehydrogenase was not significantly changed by any of the treatments.

7-Alkoxycoumarin O-Dealkylase↗
PubMed1995

Single-strand breaks in deoxyribonucleic acid in fire fighters accidentally exposed to o-nitroanisole and other chemicals.

OBJECTIVES: The aim of the study was to detect single-strand breaks in deoxyribonucleic acid (DNA) in mononuclear blood cells of fire fighters exposed to o-nitroanisole and other substances released into the environment during an accident in a chemical plant. METHODS: The level of DNA single-strand breaks in mononuclear blood cells was detected by alkaline elution. The results were compared for 16 fire fighters who worked in a contaminated area for about 8 h and two reference groups (one of fire fighters who had not worked in the contaminated area, group I, and one of persons without any apparent occupational exposure to genotoxic substances, group II). RESULTS: The mean normalized elution rate (nER) 19 d after the accident was slightly but statistically significantly (P < 0.05) higher for the exposed fire fighters [mean 1.48 +/- 95% confidence interval (95% CI) 0.21] than for reference group I (mean 1.21 +/- 95% CI 0.21) or reference group II (mean 1.17 +/- 95% CI 0.18). No statistically significant difference was found between reference groups I and II. Another analysis was performed three months after the first. The level of DNA single-strand breaks (mean nER 1.12 +/- 95% CI 0.11) was no longer increased in comparison with the levels of the reference groups. CONCLUSIONS: DNA single-strand breaks were increased in fire fighters exposed to o-nitroanisole and other substances. In comparison with the extent of DNA strand breaks found in other occupational groups the increase was only moderate. The observed decrease in DNA single-strand breaks to the reference level in exposed fire fighters three months later suggests a DNA repair mechanism for DNA single-strand breaks caused by o-nitroanisole.

Accidents↗
PubMed1994

The microsomal epoxide hydrolase has a single membrane signal anchor sequence which is dispensable for the catalytic activity of this protein.

The microsomal epoxide hydrolase (mEH) catalyses the hydrolysis of reactive epoxides which are formed by the action of cytochromes P-450 from xenobiotics. In addition it has been suggested that mEH might mediate the transport of bile acids. For the mEH it has been shown that it is co-translationally inserted into the endoplasmic reticulum. Here we demonstrate that the N-terminal 20 amino acid residues of this protein serve as its single membrane anchor signal sequence and that the function of this sequence can also be supplied by a cytochrome P-450 (CYP2B1) anchor signal sequence. The evidence supporting this conclusion is as follows: (i) the rat mEH and a CYP2B1-mEH fusion protein, in which the CYP2B1 membrane anchor signal sequence replaced the N-terminal 20 amino acid residues of mEH, was co-translationally inserted into dog pancreas microsomes in a cell-free translation system, whereas a truncated epoxide hydrolase with a deletion of the 20 N-terminal amino acid residues was not co-translationally inserted. (ii) The mEH and the CYP2B1-mEH fusion protein, but not the truncated epoxide hydrolase, were anchored in microsomes in a cell-free translation system and in membrane fractions derived from fibroblasts which expressed these proteins heterologously. These fibroblasts were also used to evaluate the significance of the mEH membrane anchor for the catalytic activity of mEH. The mEH, the truncated mEH and the CYP-EH fusion protein were found to be enzymically active. This result shows that the membrane anchor signal sequence of mEH is dispensable for the catalytic activity of this protein. However, truncated mEH was only expressed at low levels, which might indicate that this protein is unstable.

Amino Acid Sequence↗
PubMed1994

Fjord-region diol-epoxides of benzo[c]chrysene are potent inducers of micronuclei in murine bone marrow.

Vicinal diol-epoxides are the best established carcinogenic metabolites of polycyclic aromatic hydrocarbons. Numerous studies have demonstrated their high genotoxic activity in various in vitro test systems. However, in vivo mutagenicity data are not available. The fjord-region diol-epoxides of benzo[c]chrysene combine high mutagenic activity in vitro with high hydrolytic stability. They were tested for the induction of micronuclei in the bone marrow following intraperitoneal administration to NMRI mice. The anti diastereomer of the diol-epoxide enhanced the frequency of micronucleated polychromatic erythrocytes strongly (7-19-fold above the value in untreated controls) over a very wide dose range (2.5-300 mumol/kg body weight). The syn diastereomer demonstrated similar effects, but required about 5 times higher doses. The corresponding proximate mutagen, benzo[c]chrysene-trans-9,10-dihydrodiol, was only moderately active, whereas the positive control substance, benzo[a]pyrene-trans-7,8-dihydrodiol, was strongly active. The study indicates that intraperitoneally administered diol-epoxides of benzo[c]chrysene may reach the bone marrow. Therefore, it will be possible to study the influence of metabolic modulation, e.g. by enzyme induction, on the effects of these ultimate mutagens and their metabolic precursors in vivo.

Animals↗
PubMed1994

Isolation of a putative hydroxyacyl enzyme intermediate of an epoxide hydrolase.

A putative covalent, alpha-hydroxyacyl intermediate was isolated by the brief exposure of murine soluble epoxide hydrolase to its substrate. The reaction was reversed by time and blocked by competitive inhibitors. The formation of the intermediate was dependent upon the concentration of the enzyme and was increased by incubation under acidic conditions. The structure of the intermediate was supported by microchemical methods.

Acylation↗