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Z Dvorák

Publications and source records attributed to Z Dvorák.

At least 19 recordsLinked to original sources
PubMed2006

Expression and transcriptional activities of nuclear receptors involved in regulation of drug-metabolizing enzymes are not altered by colchicine: focus on PXR, CAR, and GR in primary human hepatocytes.

Recent findings show that colchicine (COL) in submicromolar concentrations downregulates the expression of major drug-metabolizing P450 enzymes in human hepatocytes. Concomitantly, the expression of pregnane X receptor (PXR) and constitutive androstane receptor (CAR) was diminished by COL, whereas expression of glucocorticoid receptor (GR) remained unaltered. A tentative mechanism is perturbation of the GR-PXR/CAR-CYP2/3 signaling cascade, resulting in restricted transcriptional activity of GR receptor by colchicine. In this work we focused on 10-demethylcolchicine (colchiceine; EIN), a structural analogue and a putative metabolite of COL that lacks tubulin-binding activity. We investigated the effects of EIN on the expression of PXR, CAR, and GR receptors in primary cultures of human hepatocytes. In contrast with the effects of COL, EIN does not alter the expression of PXR, CAR, and/or GR receptors mRNAs. In addition, EIN had no effects on transcriptional activities of PXR, CAR, and GR receptors in reporter gene assays using transfected cell lines. Considering that COL and EIN are structurally very close and differ only in their tubulin-binding activity, the data presented imply that the deleterious effects of COL on the GR-PXR/CAR-CYP2/3 cascade are primarily due to perturbation of the microtubule network. Our data support the idea of replacing COL by EIN, which is less toxic and does not interact with xenoreceptors.

Cells, Cultured↗
PubMed2006

Silybin and dehydrosilybin inhibit cytochrome P450 1A1 catalytic activity: a study in human keratinocytes and human hepatoma cells.

The flavonolignan silybin and its derivative dehydrosilybin have been proposed as candidate UV-protective agents in skin care products. This study addressed the effect of silybin and dehydrosilybin on the activity of cytochrome P450 isoform CYP1A1 in human keratinocytes (HaCaT) and human hepatoma cells (HepG2). CYP1A1 catalytic activity was assessed as O-deethylation of 7-ethoxyresorufin using fluorescence detection. Silybin and dehydrosylibin inhibited basal and dioxin-inducible CYP1A1 catalytic activity in both cell lines used. The inhibitory effect of tested compounds was more pronounced in HaCaT cells than in HepG2 cells, and dehydrosilybin was a much stronger inhibitor than silybin. Analyses on CYP1A1 human recombinant protein yielded IC(50) values of 22.9 +/- 4.7 micromol/L and 0.43 +/- 0.04 micromol/L for silybin and dehydrosilybin, respectively. Since CYP1A enzymes are some of the most prominent actors in the process of chemically induced carcinogenesis, the inhibitory activity of the flavonolignans tested against CYP1A1 favors their use as cytoprotective agents in terms of skin and hepatic metabolism. In addition, the capability of dehydrosilybin to inhibit CYP1A1 in submicromolar concentrations makes this compound a potential biological probe in CYP1A1 analyses.

Carcinoma, Hepatocellular↗
PubMed2005

Investigation of sanguinarine and chelerythrine effects on CYP1A1 expression and activity in human hepatoma cells.

Quaternary benzo[c]phenanthridine alkaloids (QBA) sanguinarine and chelerythrine exhibit a wide spectrum of biological activities whence they are used in dental care products. Recent studies indicated that cytochrome P450 CYP1A attenuates sanguinarine toxicity both in vivo [Williams, M.K., Dalvi, S., Dalvi, R.R., 2000. Influence of 3-methylcholanthrene pretreatment on sanguinarine toxicity in mice. Vet. Hum. Toxicol. 42, 196-198] and in vitro [Vrba, J., Kosina, P., Ulrichová, J., Modrianský, M., 2004. Involvement of cytochrome P450 1A in sanguinarine detoxication. Toxicol. Lett. 151, 375-387]. However, CYP1A converts sanguinarine to the products that form DNA adducts [Stiborová, M., Simánek, V., Frei, E., Hobza, P., Ulrichová, J., 2002. DNA adduct formation from quaternary benzo[c]phenanthridine alkaloids sanguinarine and chelerythrine as revealed by the 32P-postlabeling technique. Chem. Biol. Interact. 140, 231-242]. In our work we examined the effects of sanguinarine and chelerythrine on CYP1A1 expression and catalytic activity in human hepatoma cells-HepG2. Sanguinarine and chelerythrine did not affect basal and dioxin-inducible expression of CYP1A1 mRNA and protein in HepG2 cells. The enzymatic activity of CYP1A1 was assessed by the fluorescent measurement of 7-ethyxoresorufin-O-deethylase (EROD) activity. We observed a slight decrease of dioxin-induced EROD activity in HepG2 cells by sanguinarine and chelerythrine. This decrease was attributed to the inhibition of CYP1A1 catalytic activity, as revealed by enzyme kinetic studies on recombinant CYP1A1 protein. The IC50 values for the inhibition of CYP1A1 by sanguinarine and chelerythrine were 2.1 and 1.9muM, respectively. In conclusion, albeit the CYP1A modulates QBA cytotoxicity and genotoxicity, the QBA themselves do not affect CYP1A1 expression. The data indicate that studied alkaloids do not have specific cellular target and their biological effects are rather pleiotropic.

Alkaloids↗
PubMed2005

Role of microtubules network in CYP genes expression.

Superfamily of cytochrome P450 enzymes (CYPs), a distinctive enzyme system by which human body defends itself against toxic compounds, is the subject of a complex regulation process involving various mechanisms, on the levels of expression and activity. Apart from physiological factors, several patho-physiological ones such as inflammation, infection, and stress affect CYP expression. The aim of this review is to summarize the current knowledge on the role of microtubules network in the regulation of drug metabolizing CYPs. Experiments on human and animal cell models revealed that microtubules disruption severely impaired basal and inducible expression of human CYP 1A1, 2B6, 2C8, 2C9, 2C19, and 3A4, and rat CYP 1A2, 2B1, 2B2, and 3A23. Inhibition of aryl hydrocarbon receptor (AhR) and glucocorticoid receptor (GR) transcriptional activity by microtubules disarray was found to be responsible for the suppressed CYP enzymes expression. However, the mechanism by which microtubules interfering agents (MIAs) inhibit GR and AhR transcriptional activities is not fully understood yet. Several lines of evidence indicate that: i) the cell cycle, G2/M phase in particular, has an influence on AhR and GR transcriptional activity, and ii) MIAs negatively modulate GR transcriptional activity via the activation of c-Jun-N-terminal kinase. In conclusion, down-regulation of major CYP enzymes by microtubules disarray is intriguing from the mechanistic point of view and in relation to the cell differentiation.

Animals↗
PubMed2004

Speculations on the role of the microtubule network in glucocorticoid receptor signaling.

The glucocorticoid receptor (GR) is an important player in the life of a cell. This is underlined by a cohort of protein and nucleic acid structures interacting with the GR. Among many issues surrounding GR activity that are under active investigation, the role of microtubules (MTs) is still unclear. This article aims to evaluate the ayes and noes in favor of microtubule importance and then form a hypothesis on their function in GR activity.

Animals↗
PubMed2004

[Mini-invasive treatment of hemorrhoids using the Hemoron apparatus. Personal experience].

The miniinvasion therapy of hemorrhoids using the Hemoron instrument is counted among the modern methods of the out-patient treatment for the internal hemorrhoids. The aim of this work was to assess the miniinvasion therapy of the internal hemorrhoids, applying D.C. generated unipolar impulse current via endoscopy. As the result of the current applied, the arterial influx to the hemorrhoids closes down, which results in their involution.

Electrosurgery↗
PubMed2001

Cytotoxicity of natural compounds in hepatocyte cell culture models. The case of quaternary benzo[c]phenanthridine alkaloids.

The quaternary benzo[c]phenanthridine alkaloids (QBA) produce a plethora of species- and tissue-specific effects but the molecular basis of their biological activities remain mysterious. The objective of the present study was to investigate the cytotoxicity of QBA alkaloids, sanguinarine (SA), chelerythrine (CHE), fagaronine (FA), and the extract from Macleaya cordata in primary cultures of human and porcine hepatocytes. The cellular damage was assessed by the MTT assay, lactate dehydrogenase (LDH) leakage and the determination of intracellular glutathione (GSH) levels. The results are summarised as follows: (i) The alkaloids tested in doses 0.1 and 10 microM did not display statistically significant cytotoxicity for 0-3 h incubation; (ii) SA and CHE showed the dose- and time-dependent toxicity within the range 25-100 microM whereas FA was not toxic; (iii) the LDH leakage into the medium was higher for SA than for CHE, thus revealing a potent potential of SA to disturb cell-membrane integrity; (iv) after 3 h incubation with 100 microM SA/CHE, mitochondrial dehydrogenase activity (MTT assay) and the cellular GSH levels decreased to residual values of about 40% suggesting that mitochondria are unlikely to be a primary target for SA/CHE in the cell; (v) no differences were found in the response to QBA application in human vs porcine hepatocyte.

Alkaloids↗
PubMed2000

[The human hepatocyte. II. Cryopreservation].

Human hepatocytes (HH) are an optimal in vitro model for the study of xenobiotics metabolism and toxicity. However, there is still a problem of HH availability. A promising possibility for the storage of HH for later utilization is cryopreservation. Procedures and conditions used for cryopreservation of human hepatocytes and hepatocytes of other species are presented in the present paper. Compared are some biochemical parameters characterizing viability and functionality of fresh and thawed HH.

Cryopreservation↗
PubMed2000

Effect of colchicine and its derivatives on the expression of selected isoforms of cytochrome P450 in primary cultures of human hepatocytes.

The study addressed the effect of colchicine and its derivatives on the protein levels of cytochrome P450 (CYP) 1A2, 2A6, 3A4, 2C9/19, and 2E1 isoforms. Primary human hepatocyte culture was the model of choice. Levels of individual CYP isoforms were detected using immunoblotting. Colchicine caused an increase of CYP2E1 protein content, colchiceine and N-deacetylcolchiceine induced isoforms CYP2C9, 2E1 and 3A4 whereas colchicoside induced CYP2C9 and 2E1. The levels of CYP1A2 and 2A6 were unaffected by any of tested compounds. Demecolcine and 3-demethylcolchicine had no effect on any studied P450 isoform. Since colchicine is an exclusive substrate of CYP3A4 whereas it induces CYP2E1, there is a suspicion rather at protein stabilization than at gene induction concerning induction origin.

Cells, Cultured↗
PubMed1999

[The human hepatocyte: I. A model for studying metabolism and toxicity of xenobiotics].

Isolated human hepatocytes have become one of the most attractive experimental approaches to the study of the specific metabolic functions of the human liver, interactions between liver cells and infectious agents, and metabolism and pharmacotoxicity of drugs. Particularly the primary cell culture model provides an in vitro system for investigating specific mechanisms in a precisely controlled conditions. In the present paper the legislative and ethical problems concerning availability of human liver samples, techniques developed for isolation, preservation and cultivation of hepatocytes are discussed. In addition, a comparison of human and rat hepatocyte models in the study of the metabolism and cytotoxicity of taxol is reviewed.

Animals↗
PubMed1990

Mechanical engineering problems in preserving biological objects by temperature lowering.

Analysis of dangers caused by mechanical refrigerating and liquid nitrogen systems used for low temperature preserving of biological material and safety measures to be adopted. Hazards are caused by moving or protruding parts of the machinery, its hot parts, noise and vibration, work in cold rooms, possible destruction of pressure vessels, refrigerant inflammation or explosion, breathing the refrigerant or its decomposition products, direct contact of the refrigerant with the skin or mucous tissues, depletion of stratospheric ozone or contamination of food-stuffs.

Equipment and Supplies↗
PubMed1986

Evaluation of protein mixtures in adults on the basis of scoring pattern deduced from Kofrányi's results.

Minimum protein requirements for adult males found by Kofrányi in testing protein mixtures were compared with amino acid scoring pattern in which total non-essential amino acids were also included. For protein mixtures composed of different ratios of animal and plant proteins the least minimum of protein required corresponded to the maximum of the courses of chemical score when non-essential amino acids, lysine, sulfur amino acids or tryptophan respectively were taken as limiting. Particular comparisons indicate that total non-essential amino acids ought to be included in amino acid scoring pattern.

Adult↗
PubMed1986

[Acid-base equilibrium in the blood in cattle in relation to the incidence of DFD (dark, firm, dry) meat].

We studied changes in the parameters of the acid-base balance of the blood in cattle before animal transport to the slaughterhouses, after transport, after stunning and before bleeding. At the same time we studied the occurrence of DFD meat by measuring the pH value of the muscle and by determining the meat color. We found out that the disorder of acid-base balance was less manifest, with respect to the large capacity of the buffer systems of the blood, than the changes caused by glycogenolysis in the muscle.

Acid-Base Equilibrium↗
PubMed1975

[Loss of nutritional value of proteins in meat products caused by thermal processing].

In certain cases, the proteins in meat products may be damaged during thermal processing to such an extent that their nutritional value is reduced. This might be caused above all by a loss in the availability of the essential and semi-essential amino-acids. Cooking of the meat produces no marked change in the availability of the amino acids. More marked changes are observed at temperatures needed for sterilization. The availability of cystine is in most cases affected by the length and temperature of sterilization, which is due to a reduction of its digestibility and to its degradation. The extent of the cystine degradation follows from the difference from the average reduction of the availability of the other amino acids. Since the sulphur-containing amino acids in meat proteins are limiting ones, this loss results in a reduction of the nutritional value of the proteins. Apart from cystine, lysine may also become less available if greater amounts of polysaccharides are present. The reduction of the availability can be so marked that lysine may be considered to be a limiting amino acid. In these cases, lysine is essential for the nutritional value of the proteins in such products.

Amino Acids↗