Biomedical subjects

M Monshouwer

Publications and source records attributed to M Monshouwer.

24 records · Page 2Linked to original sources

PubMed1995

The antibiotic tiamulin is a potent inducer and inhibitor of cytochrome P4503A via the formation of a stable metabolic intermediate complex. Studies in primary hepatocyte cultures and liver microsomes of the pig.

Tiamulin is a semisynthetic antibiotic frequently used in agricultural animals. The drug has been shown to produce clinically important--often lethal--interactions with other compounds that are simultaneously administered. To explain this, it has been suggested that tiamulin selectively inhibits oxidative drug metabolism via the formation of a cytochrome P450 metabolic intermediate complex. The aim of the present study was to provide further support for this hypothesis. When hepatic microsomes and cultured primary pig hepatocytes were incubated with tiamulin, a maximum in the absorbance spectrum at 455 nm was observed, which disappeared after adding KFe(CN)6. When hepatocytes were incubated with tiamulin for 72 hr, cytochrome P450 content and cytochrome P4503A apoprotein levels were increased. Tiamulin strongly inhibited and concentration dependently inhibited the hydroxylation rate of testosterone at the 6 beta-position in both microsomes and hepatocytes, and the microsomal N-demethylation rate of ethylmorphine. Other testosterone hydroxylations were inhibited to a lesser extent or not affected. The relative inhibition of the hydroxylation of testosterone at the 6 beta-position was more pronounced in microsomes from rifampicin- and triacetyloleandomycin-treated pigs. The results indicate that cytochrome P450 complex formation can at least partly explain the interactions observed with tiamulin. Tiamulin seems to be a strong, probably selective, inhibitor of the cytochrome P4503A subfamily and an interesting tool for further research.

Animals↗

PubMed1995

Infection (Actinobacillus pleuropneumoniae)-mediated suppression of oxidative hepatic drug metabolism and cytochrome P4503A mRNA levels in pigs.

The effect of Actinobacillus pleuropneumoniae infection, a well-characterized pig infection model, on both phase I (oxidative) and phase II (conjugative) microsomal enzyme activities was investigated in castrated male conventional pigs. A. pleuropneumoniae infection resulted after 24 hr in a significant suppression of 33% or more of all oxidative enzyme activities determined. After 40 hr, the activities were still suppressed, but did not differ from the results after 24 hr. On the contrary, all glucuronosyltransferase activities measured were not affected by A. pleuropneumoniae infection after both 24 and 40 hr. To elucidate further the mechanism of the suppression of oxidative enzyme activities, analysis of mRNA were conducted by dot-blot analysis using a human cytochrome P4503A4 cDNA probe. The results indicated that A. pleuropneumoniae infection suppressed oxidative enzyme activities. The reduction in cytochrome P4503A activity, specific for 6 beta-hydroxylation of testosterone is at a pretranslational level as measured by a decrease in the amount of mRNA.

Actinobacillus Infections↗

PubMed1994

A reversed-phase high-performance liquid chromatographic method for the determination of Clanfenur in rat and human plasma.

A selective and specific high-pressure liquid chromatographic (HPLC) method for the simultaneous assay of Clanfenur and its metabolites in biological fluids of interest has been developed which is suitable for routine analysis, using micro volumes (0.1 ml) of plasma samples only. After protein precipitation the extract is analysed by reversed-phase HPLC with UV detection. Excellent recovery, linearity, accuracy and precision (less than 5% for plasma) are achieved by the assay which is able to quantify Clanfenur and its metabolites in plasma at concentrations between 0.025 and 5.0 mg l-1.

Animals↗

PubMed1994

Dose-dependent pharmacokinetic interaction between antipyrine and paracetamol in vivo and in vitro when administered as a cocktail in pig.

1. The pharmacokinetic interactions between paracetamol (PA) and antipyrine (AP) were studied in pigs in order to investigate the usefulness of this combination for the simultaneous assessment of oxidative and conjugative metabolism. 2. When both drugs were given at a dose of 5 mg/kg, AP plasma clearance was reduced from 2.22 to 0.96 lh-1 kg-1. PA clearance was not changed in comparison with control values. 3. At a dose of 2 mg/kg no pharmacokinetic interaction between the two drugs was observed. 4. The only oxidative AP metabolite found in urine was 4-hydroxyantipyrine (4-OHA). It accounted for 80% of the dose and, like PA, it was completely glucuronidated. 5. The glucuronidation of PA has been studied in vitro in pig liver microsomes. The apparent Km value for PA glucuronidation was 40 mM with a Vmax = 54 nmol min-1 mg protein-1. To determine if 4-OHA and PA competed for the same UDP-glucuronosyl-transferase, the effect of 4-OHA and AP on PA glucuronidation was studied. It appeared that 4-OHA was a competitive inhibitor with a Ki app = 0.07 microM, whereas AP had no effect. 5. Results suggest a dose-dependent interaction between AP and PA, which may be due to competition at the level of glucuronidation. Therefore, the usefulness of AP and PA in vivo in a cocktail for metabolism studies is limited.

Acetaminophen↗

PubMedDate not supplied

The use of genetically engineered cells for assessing CYP2D6-related polymorphic effects.

As an example of advanced testing in the field of metabolism in an industrial environment, the introduction of some novel approaches, including the use of genetically engineered cell lines for assessing CYP 2D6-related polymorphic effects is illustrated. In this paper, it is demonstrated that novel in vitro test systems can be developed by using these genetically engineered cell lines for evaluating the potential risks associated with proprietary drugs (especially if their metabolism depends to a high extent on CYP 2D6). Moreover, it is demonstrated that, by the use of these in vitro methods, issues such as polymorphism, for which no animal models are available, can be assessed in such a way that predictions can be made on adverse effects which, up to now, could only be detected during clinical trials. Through the use of these new biotechnological in vitro metabolism models, clinically relevant data can be obtained for a scientifically-based human risk assessment, and animal use can be reduced.

Animal Testing Alternatives↗

PubMedDate not supplied

Participation of beta-adrenergic receptors on macrophages in modulation of LPS-induced cytokine release.

For several years it is known that beta-adrenergic receptor agonists have anti-inflammatory effects. However, little is known about the role of beta-adrenergic receptors on macrophages in the modulation of cytokine production by beta-agonists during inflammation. In this study, the presence of beta-receptors on PMA-differentiated U937 human macrophages, and the participation of these receptors in the modulation of LPS-mediated cytokine production by beta-agonists was investigated. Total beta-receptor expression on undifferentiated (monocyte) and PMA-differentiated U937 cells was established using receptor binding studies on membrane fractions with a radio ligand. The expression of beta-receptors proved to be significantly lower on monocytes than on macrophages, additionally a predominant expression of beta 2-receptors was found. Production of the cytokines TNF-alpha, IL-6, and IL-10 by LPS-stimulated differentiated U937 cells was measured in time. Peak concentrations for TNF-alpha, IL-6 and IL-10 occurred at 3, 12 and 9 hrs, respectively. When differentiated U937 cells were incubated with both LPS and the beta-agonist clenbuterol the production of TNF-alpha and IL-6 was significantly reduced. However the production of IL-10 was increased. To study the mechanism of modulation of cytokine production in more detail, U937 macrophages were incubated with LPS/clenbuterol in combination with selective beta 1- and beta 2-antagonists. These results indicated that the beta 2- and not the beta 1-receptor is involved in the anti-inflammatory activity of clenbuterol.

Adrenergic beta-Agonists↗