Showing papers in "Biomedical Papers-olomouc in 2005"

Silybin and silymarin - new effects and applications

Abstract: This article aims to review critically literature published mainly within this millennium on the new and emerging applications of silymarin, the polyphenolic fraction from the seeds of Silybum marianum and its main component silybin. Silymarin and silybin used so far mostly as hepatoprotectants were shown to have other interesting activities as e.g., anticancer and canceroprotective. These activities were demonstrated in a large variety of illnesses of different organs as e.g., prostate, lungs, CNS, kidneys, pancreas and others. Besides the cytoprotective activity of silybin mediated by its antioxidative and radical-scavenging properties also new activities based on the specific receptor interaction were discovered--e.g., inhibition and modulation of drug transporters, P-glycoproteins, estrogenic receptors, nuclear receptors and some others. New derivatives of silybin open new ways to its therapeutic applications. Pharmacology dealing with optically pure silybin diastereomers may suggest new mechanisms of its action.

Myrrh--Commiphora chemistry.

Abstract: Myrrh and opopanax has been used throughout history in incense and as a perfume. Since Bible times it has been used for the treatment of wounds. The first attempts to identify content compounds were almost 100 years ago. In this review we discuss the present state of knowledge in the chemistry of substances of Commiphora spp.

Increased level of advanced oxidation products (AOPP) as a marker of oxidative stress in patients with acute coronary syndrome.

Abstract: Oxidative stress impairs endothelial function and may play an important role in the pathogenesis of acute cardiovascular diseases. Advanced oxidation protein products (AOPP) were proposed as one of the possible markers of oxidative injury, which originates under oxidative and carbonyl stress and increase global inflammatory activity. The present study was undertaken to compare AOPP concentrations in a control group of healthy individuals without ICHS (I), patients with stable angina pectoris (II), patients with acute coronary syndrome over 48 hours without ST elevations (III), and patients with ST elevation myocardial infarction (IV). Coronaronary angiography, risk factors and anamnestic data were analyzed. We examined 73 probands with signs of myocardial ischemia, mean age of 61.5 years (64% males) subjected to coronarography and 21 healthy individuals. No significant difference was found between venous blood and coronary samples, or between infarction and non-infarction arteries in the group IV. AOPP concentrations in healthy individuals in the group I (82.9 +/- 29.3 mmol/l) did not differ significantly from patients in group II (89.6 +/- 26.7 mmol/l) and group III (112.3 +/- 54.6 mmol/l). A significant difference in AOPP values was found between the groups I and IV, and between the groups II and IV (82.9 +/- 29.3 mmol/l vs. 125.8 +/- 101 mmol/l, p = 0.02, and 89.6 +/- 26.7 mmol/l vs. 125.8 +/- 101 mmol/l, p = 0.02). No correlations were found between AOPP and body mass index (BMI), nicotinism, left ventricular ejection fraction, parameters of glucose and lipid metabolism. ROC analysis revealed that AOPP concentrations of 89 mmol/l had 64% sensitivity and 71% specificity for revealing an acute coronary syndrome (AUC 0.65, 95% CI 0.55-0.80). AOPP are significantly increased in patients with acute coronary syndromes with ST segment elevation, but also tend to increase in patients with non-ST elevation myocardial infarction. Our observations suggest that AOPP may be used as a marker of oxidative stress and as a prognostic factor for severe forms of cardiovascular disease. A cut-off value of 89 mmol/l can be used with 64% sensitivity and 71% specificity for revealing acute coronary syndrome.

Dual role of cathepsin D: ligand and protease.

Abstract: Cathepsin D is peptidase belonging to the family of aspartic peptidases. Its mostly described function is intracellular catabolism in lysosomal compartments, other physiological effect include hormone and antigen processing. For almost two decades, there have been an increasing number of data describing additional roles imparted by cathepsin D and its pro-enzyme, resulting in cathepsin D being a specific biomarker of some diseases. These roles in pathological conditions, namely elevated levels in certain tumor tissues, seem to be connected to another, yet not fully understood functionality. However, despite numerous studies, the mechanisms of cathepsin D and its precursor's actions are still not completely understood. From results discussed in this article it might be concluded that cathepsin D in its zymogen status has additional function, which is rather dependent on a "ligand-like" function then on proteolytic activity.

Hair analysis for drugs of abuse. Plausibility of interpretation.

Abstract: Over more than 20 years hair analysis for drugs has been gaining increasing attention and recognition in various toxicological fields as preemployment and employment screening, forensic sciences, doping control of banned substances, clinical diagnostics in health problems. Hair analysis for drugs can expand the toxicological examination of conventional materials and thus contribute with additional important information to the complex evaluation of a certain case. Hair is a unique material for the retrospective investigation of chronic drug consumption, intentional or unintentional chronic poisoning in criminal cases, gestational drug exposure or environmental exposure to pollutants and adulterants and with specific ultrasensitive procedures allow to demonstrate even a previous single dose administration in a very low amount. Assuming the ideal hair steady and uniform growth, segmental hair analysis can provide the information about the time course of the substance use or exposure. However, the physiological background of hair growth, mechanisms of drug incorporation are not simple, not yet understood in full details and need not be evaluated exactly in all cases. The hair sampling, storage, sample preparation, analytical performance themselves are also very important for final results. Different laboratory attitudes can produce different results. The full information on circumstances of the case examined must be taken into account during interpretation. The pitfalls in hair analysis should be known and avoided to assure the responsible and correct interpretation of laboratory results adequate to an individual case.

Antimutagenic effect of phenolic acids.

Abstract: In the present study, the Salmonella typhimurium tester strain TA 100 was used in the plate-incorporation test to examine the antimutagenic potential of caffeic, ferulic and cichoric acids extracted from plant species of genera Echinacea (L) Moench, as well as of another phenolic acids, on 3-(5-nitro-2-furyl)acrylic acid (5NFAA) and sodium azide mutagenicity. All tested compounds possess antimutagenic activity. In the case of 5NFAA, the antimutagenic potency of tested compounds was in the order of gallic acid > ferulic acid > caffeic acid > syringic acid > vanillic acid. The mutagenic effect of sodium azide was inhibited by tested phenolic acids by about 20-35 %. The most effective compound, gallic acid inhibits this effect by 82 % in the concentration of 500 mug/plate. The only exception from favourable properties of tested phenolic acids is cichoric acid, which in the contrary significantly increased the mutagenic effect of 5NFAA.

Heat shock proteins in autoimmune diseases.

Abstract: Heat shock proteins (hsp's) are among the most conserved proteins in evolution. They have been identified as important pathogen-related antigens as well as autoantigens suitable for construction of novel vaccines. The high evolutionary homology of hsp's has raised the question about the safety of such vaccines. Experimental and clinical observations have confirmed that hsp proteins are involved in the regulation of some autoimmune disease such as autoimmune arthritis, type 1 diabetes mellitus, atherosclerosis, multiple sclerosis, and other autoimmune reactions. It has been shown in experimental animals that some hsp proteins (especially hsp60, hsp70, and hsp10) can either induce or prevent autoimmune reactions depending on the circumstances. This article discusses the involvement of hsp proteins in the etiology of autoimmune diseases and it presents promising experimental data on the effects of immunization with hsp proteins in the prevention and therapy of autoimmune diseases.

Comparison the cytotoxicity of hydroxyapatite measured by direct cell counting and mtt test in murine fibroblast nih-3t3 cells

Abstract: The worldwide growing interest to biomaterials over the last years results from their irreplaceable role in medical clinic. Hydroxyapatite is used in bone reconstruction because of its similar chemical structure compared to the inorganic composition of human bone and it is basic building component of many newly prepared biomaterials. In this study, we evaluated cytotoxic/antiproliferative activity of hydroxyapatite extract using murine fibroblast cell line NIH-3T3 and two in vitro different cytotoxic assays: growth inhibition assay and MTT assay. Hydroxyapatite extract after 72 h of incubation manifested the significant in vitro cytotoxic/antiproliferative effect only at the highest concentration tested (100 %). The antiproliferative effect of hydroxyapatite extract at the other concentrations tested (75 %, 50 %, 25 %, 10 %, 5 % and 1 %) was directly proportional to the concentration and the time of influence. The inhibition of cell proliferation was 86.8 - 0 %. The sensitivity of cell growth inhibition assay (direct counting of viable cells) to the extract influence was higher than that of MTT test.

Differentiation of tumours of ductal and lobular origin: I. Proteomics of invasive ductal and lobular breast carcinomas.

Abstract: The vast majority of invasive breast tumors are ductal and lobular breast carcinomas. Despite the many similarities, some clinical follow-up data and the patterns of metastases suggest that these histological subtypes of breast cancer are biologically distinct. Few papers, however, describe immunohistochemical markers useful for differentiation of these carcinomas. Many investigations suggest that E cadherin protein expression is lost in lobular but not in ductal carcinoma. The absence of E-CD, as a partial loss of epithelial differentiation, may account for the extended spread of lobular carcinoma in situ and the peculiar diffuse invasion mode of invasive lobular carcinoma. Some investigations report the significance of E-CD associated proteins alpha-, beta-, gamma-catenin expression, as well as the usefulness of cytokeratins 5, 6, 8, 7 and thrombospondin in differentiating histological types of breast invasive carcinomas. Several reports have suggested the possibility that invasive ductal and lobular cancers differ with respect to expression of antigens involved in proliferation and cell cycle regulation. It has been shown that vascular endothelial growth factor expression, also the expression of maspin, a tumour suppressor gene product, is higher in ductal, than in lobular carcinoma. Expression of NKX3.1, a member of the NK-class of homeodomain, is highly restricted and is found primarily in lobular carcinoma. Some histological and immunohistochemical characteristics of pleomorphic lobular carcinoma are also discussed.

New quaternary pyridine aldoximes as casual antidotes against nerve agents intoxications.

Abstract: In this work, the ability of four newly synthesized oximes - K005 (1,3-bis(2-hydroxyiminomethylpyridinium) propane dibromide), K027 (1-(4-hydroxyiminomethylpyridinium)-3-(4-carbamoylpyridinium) propane dibromide), K033 (1,4-bis(2-hydroxyiminomethylpyridinium) butane dibromide) and K048 (1-(4-hydroxyiminomethylpyridinium)-4-(4- carbamoylpyridinium) butane dibromide) to reactivate acetylcholinesterase (AChE, EC 3.1.1.7) inhibited by nerve agents is summarized. Reactivation potency of these compouds was tested using standard in vitro reactivation test. Tabun, sarin, cyclosarin and VX agent were used as appropriate testing nerve agents. Rat brain AChE was used as a source of the enzyme. Efficacies of new reactivators to reactivate tabun-, sarin-, cyclosarin- and VX-inhibited AChE were compared with the currently used AChE reactivators (pralidoxime, obidoxime and HI-6). Oxime K048 seems to be promising reactivator of tabun-inhibited AChE. Its reactivation potency is significantly higher than that of HI-6 and pralidoxime and comparable with the potency of obidoxime. The best reactivator of sarin-inhibited AChE seems to be oxime HI-6. None of the new AChE reactivators reached comparable reactivation potency. The same results were obtained for cyclosarin-inhibited AChE. However, oxime K033 is also potent reactivator of AChE inhibited by this nerve agent. In the case of VX inhibition, obidoxime and new oximes K027 and K048 seem to be the best AChE reactivators. None from the currently tested AChE reactivators is able to reactivate AChE inhibited by all nerve agents used and, therefore, the search for new potential broad spectrum AChE reactivators is needed.

Universal primers for detection of common bacterial pathogens causing prosthetic joint infection.

Abstract: The diagnosis of low grade prosthetic joint infection is difficult and time consuming. Nested-PCR for universal bacterial DNA segments detection of "orthopaedic" bacteria was tested in a laboratory setting. This method is based on amplification of the 16S bacterial ribosomal RNA coding sequences. 11 species of the most frequent bacterial pathogens (Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus pyogenes, Streptococcus agalactiae, Enterococcus faecium, Enterococcus faecalis, Klebsiella pneumoniae, Escherichia coli, Proteus mirabilis, Pseudomonas aeruginosa, Serratia marcescens) involved in prosthetic joint infections were studied. All could be detected rapidly and sensitively by this method.

The efficacy of glucosamine and chondroitin sulfate in the treatment of osteoarthritis: are these saccharides drugs or nutraceuticals?

Abstract: This review summarizes recent knowledge on the efficacy of glucosamine (GS) and/or chondroitin sulfate (CS) in the therapy of mild to moderate osteoarthritis (OA). OA, the most common joint disease is a significant source of disability, quality of life impairment and a considerable burden to any health care system. In the Czech Republic, glucosamine sulfate (GS) and chondroitin sulfate (CS) are available both as prescription drugs and as food supplements. Based on available data both are useful in the earlier stages of OA when combined with other modalities such as weight loss and exercises. They appear to relieve pain and improve range of the joint motion. In addition, they also display mild anti-inflammatory effects. However, controversy still exists over their ability to change significantly the natural history of the osteoarthritic joint. This effect is not easy to demonstrate for any other treatment modalities apart from joint replacement. Monitoring the cure efficacy by X-ray has been recently criticised and hence future techniques are anticipated for this reason. Further, long-term oral administration is required to obtain slightly increased levels of GS and/or CS in human blood. Both reviewed saccharides are well tolerated with negligible adverse reactions. In conclusion, the authors suggest that GS and CS should be classified as food supplements only.

In vitro testing of gentamicin-vancomycin loaded bone cement to prevent prosthetic joint infection.

Abstract: Sepsis is a greatly feared complication of total joint arthroplasty. One key question is how to prevent perioperative bacterial adherence, and therefore the potential for infectious complications. The objective of our study was to appraise the emerging capacity of staphylococcal survival on prosthetic materials and to analyze the in vitro effects of gentamicin and vancomycin loaded polymethylmethacrylate (PMMA) cement on bacterial adherence and growth. Hospital acquired staphylococcal strains were systematically inoculated on four orthopedic materials (ultrahigh molecular weight polyethylene, PMMA without antibiotic, commercially produced PMMA loaded with gentamicin, and manually mixed PMMA loaded with gentamicin and vancomycin). Staphylococci were identified using culture and biochemical tests. The inoculated material was allowed to incubate in a liquid broth growth media and subsequently prepared for scanning electron microscopy and bacterial growth quantification. Materials without antibiotics showed evidence of staphylococcal growth. PMMA loaded with only gentamicin grew methicillin-resistant Staphylococcus aureus. Gentamicin-vancomycin loaded PMMA completely inhibited any bacterial growth. Low-dose gentamicin-vancomycin loaded PMMA prevents staphylococcal colonization better than commercially manufactured PMMA loaded with gentamicin. We recommend this combination in high-risk procedures and revision surgeries requiring bone cement.

Antiproliferative activity of berberine in vitro and in vivo.

Abstract: Berberine, an isoquinoline plant alkaloid acted cytotoxically in vitro on tumour cell lines B16. Its anticancer activity in vivo was studied with the transplanted B16 line in the range of doses from 1 mg/kg to 10 mg/kg. The significant reduction of tumor volume was observed on day 16 at doses of 5 and 10 mg/kg. The dose of 1 mg/kg stimulated the tumor mass, but other tested concentration, 5 and 10 mg/kg, reduced the tumor weight.

Serum leptin in the development of insulin resistance and other disorders in the metabolic syndrome.

Abstract: The metabolic syndrome mostly represented by obesity and hyperinsulinaemia connected with insulin resistance, presents the main mechanism in the pathogenesis of cardiovascular disease. The aim of this study was to analyze the interrelations between several metabolic variables (including leptin) and factors related to insulin resistance in groups of both normal and non-diabetic hyperlipemic postmenopausal women and men of appropriate age, and to attempt to elucidate the gender differences. Two groups of patients (20 men, 20 women) with hypertriglyceridemia were compared with 30 individuals (10 men, 20 women) with normal serum triacylglycerols. Fasting serum leptin concentration, lipid parameters (triacylglycerols, HDL cholesterol, LDL cholesterol) and BMI were measured and compared with changes in insulin parameters influencing insulin resistance (HOMA IR, insulin, intact proinsulin, C-peptide). Statistical analysis was performed using SAS/STAT software including unpaired Student's t-test, Kolmogorov-Smirnov's test, Spearman's rank-order correlation and multiple regression analysis. In men, the insulin sensitivity correlates with leptin only. In women insulin sensitivity is markedly influenced by a complex of factors: leptin and lipid parameters. Increased insulin resistance in men is followed mainly by the increased correlations between leptin, HOMA IR and insulin parameters. In women correlations between leptin, HOMA IR and insulin parameters were smaller, but the inverse correlation with HDL cholesterol was stronger. In postmenopausal women and also in men, serum leptin concentration contributes to insulin resistance. However in women the effect of increase in serum triacylglycerols in contribution of insulin resistance seems to be more dominant.

TNF-α IN THE DEVELOPMENT OF INSULIN RESISTANCE AND OTHER DISORDERS IN METABOLIC SYNDROME

Abstract: Insulin resistance and obesity are very frequent disorders and are described as the dominant risk factors for cardiovascular disease. The aim of this study was to analyze the interrelations between several metabolic variables (including TNF-α) and factors related to insulin resistance in groups of both normal and hyperlipidemic postmenopausal women and men of appropriate age, and to attempt to elucidate the gender differences. The study was carried out on 70 out-patients of the Metabolic Center. From these, 40 patients (20 men and 20 women) were selected with mild hyperlipidemia. Two other groups (10 men and 20 women) with approximately normal serum lipids parameters were taken as “controls”. In hyperlipidemic women the mean serum concentration of the TNF-α was no different from that in the control group in spite of the fact that values of HOMA IR, insulin, proinsulin and lipid parameters increased significantly. In hyperlipidemic men we have found the decrease in TNF-α in comparison with the control group. In all four groups the statistical analysis showed correlations between metabolic parameters (including TNF-α) and parameters related to insulin resistance. Also differences in relation to the gender have been found. Multiple regression analysis demonstrated the important role of TNF-α in the regulation of both the insulin resistance and in the secretion of insulin in women. In men, BMI and HDL-cholesterol played a dominant role, while the role of TNF-α seemed to be minimal.

Relation between pH and the strong ion difference (SID) in body fluids.

Abstract: Acid-base balance evaluation according to the Henderson-Hasselbalch equation enable us to assess the contribution of respiratory (pCO2) and/or non-respiratory (metabolic, HCO3(-)) components to the acid-base balance status. A new approach to acid-base balance evaluation according to Stewart-Fencl, which is based on a detailed physical-chemical analysis of body fluids shows that metabolic acid-base balance disorders are characterized not only by [HCO3(-)]. According to this concept independent variables must be taken into an account. The abnormality of concentration of one or more of the independent variable(s) determines the pH of a solution. The independent variables are: 1. strong ion difference (SID); 2. total concentration of nonvolatile weak acids [A(tot)]; 3. in agreement with the Henderson-Hasselbalch concept also pCO2. Traditional evaluation of acid-base balance disorders is based on the pH of body fluids (though pH may be within normal range if several acid-base balance disturbances are present). In order to maintain this view and simultaneously to respect the Stewart-Fencl principle, we invented a new equation, which uses only the independent variables to define the pH of body fluids. This analysis shows that for a given value of pCO2, the pH of body fluids is determined by a difference between SID and [A(tot)-]. pH = 6.1 + log((SID - [A(tot)-])/(0.03pCO2)) or in itemized form: pH = 6.1 + log((([Na+] + [K+] + [Ca2+] + [Mg2+] - [Cl-] - [UA-]) - (k1[Alb] + k2[P(i)]))/(0.03 x pCO2)). Evaluation of the individual components of this equation enables us to detect, which of the independent variable (or a combination of independent variables) deviates from the normal range and therefore which one or ones is a cause of the acid-base balance disorder. At the end of this paper we give examples of a practical application of this equation.

Effect of coenzyme Q(10) supplementation in the rat model of adjuvant arthritis.

Abstract: Adjuvant arthritis (AA) is a model of chronic inflammation induced by Mycobacterium butyricum and characterized by similar pathophysiological and pathobiochemical changes as rheumatoid arthritis (RA) in humans. In this study the antirheumatic activity of coenzyme Q10 supplementation was tested not only as to its capability to suppress the inflammation edema of the hind paw and to improve the body weight of the arthritic animals, but also to improve so important biochemical parameters as markers of inflammation and oxidative stress, and of mitochondrial bioenergetics. Despite the unfavorable effects on the rheumatic processes observed by monitoring biometric parameters (hind paw volume, relative body weight, relative weight of spleen), a significant protective effect was observed on the level of mitochondrial energetic and antioxidant disbalance. This finding speaks in favor of CoQ10 supplementation in rheumatic patients, presumably as combinatory therapy with classical antirheumatics, e.g. NSAIDs.

Matrix metalloproteinases and their function in myocardium.

Abstract: A significant number of myocardial diseases are accompanied by increased synthesis and degradation of the extracellular matrix (ECM) as well as by changed maturation and incorporation of ECM components. Important groups of enzymes responsible for both normal and pathological processes in ECM remodeling are matrix metaloproteinases (MMPs). These enzymes share a relatively conserved structure with a number of identifiable modules linked to their specific functions. The most important function of MMPs is the ability to cleave various ECM components; including such rigid molecules as fibrillar collagen molecules. The amount and activity of MMPs in cardiac tissue are regulated by a range of activating and inhibiting processes. Although MMPs play multifarious roles in many myocardial diseases, here we have focused on their function in ischemic cardiac tissue, dilated cardiomyopathy and hypertrophied cardiac tissue. The inhibition of MMPs by means of synthetic inhibitors seems to be a promising strategy in cardiac disease treatment. Their effects on diseased cardiac tissue have been successfully tested in several experimental studies.

Soluble cell adhesion molecules s-VCAM-1 and s-ICAM-1 in subjects with familial combined hyperlipidemia.

Abstract: intima-media thickness/Atherosclerosis Familial combined hyperlipidemia (FCH) is the most common familial hyperlipidemia with a high risk for early atherosclerosis. The aim of this study was to compare levels of soluble intercellular cell adhesion molecule 1 (s-ICAM-1) and soluble vascular cell adhesion molecule 1 (s-VCAM-1) in asymptomatic members of FCH families with healthy controls and to determine the relation between s-ICAM-1, s-VCAM-1 and risk factors accompanying FCH. We also investigated the association between adhesion molecules and the intima-media thickness (IMT) of the common carotid artery, a recognized morphological marker of early atherosclerosis. 82 members of 29 FCH families were divided into the 2 groups: HL (probands and hyperlipidemic first-degree relatives, n = 47) and NL (normolipidemic first-degree relatives, n = 35). The control groups – HL-C (n = 20) and NL-C (n = 20) – consisted of sex- and age–matched healthy individuals. Hyperlipidemic members had significantly higher concentration of s-ICAM-1 (633.7 ± 169.6 ng/ml versus 546.2 ± 155.9 ng/ml, p < 0.05). The elevation of s-VCAM-1 was not significant (880.8 ± 202.9 ng/ml versus 826.5 ± 174.6 ng/ml, N.S.). Levels of s-ICAM-1 and of s-VCAM-1 in normolipidemic relatives were not significantly different from the control group (530.8 ± 113.9 ng/ml versus 530.0 ± 101.0 ng/ml and 860.2 ± 265.7 ng/ml versus 822.1 ± 197.0 ng/ml respectively). There was a significant correlation between s-ICAM-1 and apoB (r = 0.42; p < 0.01) in hyperlipidemic subjects and between s-ICAM-1 and proinsulin (r = 0.54; p < 0.01) in normolipidemic subjects. S-ICAM-1 correlated with IMT (r = 0.32; p < 0.05) in all members of FCH families. The increase of s-ICAM-1 in asymptomatic hyperlipidemic members of FCH families reflects their high cardiovascular risk. The positive association between s-ICAM-1 and IMT could indicate s- ICAM-1 as a potential predictor of atherosclerosis manifestation.

Percutaneous osteosynthesis versus cast immobilisation for the treatment of minimally and non-displaced scaphoid fractures. Functional outcomes after a follow-up of at least 12 month.

Abstract: The authors compare the functional outcome of 26 patients successfully treated by percutaneous osteosynthesis and 24 patients successfully treated by a short-arm thumb cast for the acute scaphoid fracture after a follow-up of at least 12 months. The patients treated by percutaneous osteosynthesis showed a significantly better range of wrist motion at the time of re-examination. Differences in persistent complaints and in grip strength compared to the uninjured wrist between both groups were statistically insignificant yet slightly in favour of the surgically treated patients. The higher suitability of percutaneous osteosynthesis is further supported by the significantly lower failure rate compared to conservative treatment. Based on these results, percutaneous osteosynthesis seemed to be the more favourable method of treatment of minimally and non-displaced scaphoid fractures than conservative treatment.

Antitumor drug ellipticine inhibits the activities of rat hepatic cytochromes p450

Abstract: Ellipticine is a potent antineoplastic agent, whose mode of action is considered to be based mainly on DNA intercalation and/or inhibition of topoisomerase II. Recently, we found that ellipticine also forms the cytochrome P450 (CYP)-mediated covalent DNA adducts. Here, we study the effect of ellipticine on CYP enzymes in rat hepatic microsomes, studying its binding to the enzymes and its potential to inhibit the CYP activities measured with their selective substrates. Although ellipticine was reported to be a selective and strong inhibitor of CYP1A1/2, we found that its inhibitory potential is non-specific. Ellipticine is the most potent inhibitor for CYP3A-dependent 6beta-hydroxylation of progesterone, followed by CYP1A1/2-dependent ethoxyresorufin O-deethylation and CYP2B-mediated pentoxyresorufin O-depentylation. Lower inhibition was detected for 1'-hydroxylation of bufurarol, 21-hydroxylation of progesterone and 6-hydroxylation of chlorzoxazone catalyzed by CYP2D, CYP2C and CYP2E1, respectively. Ellipticine binds to several CYPs of rat hepatic microsomes. The binding titration of ellipticine typically give reverse type I spectrum with CYPs in rat hepatic microsomes. The results indicate that inhibition of CYPs by ellipticine cannot be explained only by its differential potency to bind to individual CYPs.

High adiponectin and TNF-alpha levels in moderate drinkers suffering from liver steatosis: comparison with non drinkers suffering from similar hepatopathy.

Abstract: Moderate alcohol consumption is associated with increased insulin sensitivity and a reduced risk for type 2 diabetes. An important endogenous mediator of insulin sensitivity is adiponectin (AN), an adipokine that displays numerous antiatherogenic, antidiabetogenic and antiinflammatory effects. Recently, acute increase in alcohol consumption has been shown to be associated with increase in plasma adiponectin and, concomitantly, insulin sensitivity. Whether chronic alcohol consumption predicts an increase in plasma AN and whether this is independent of adiposity, markers of liver dysfunction, and plasma adipokines such as tumor necrosis factor (TNF)-α is not known. We, therefore, investigated these relationships in 75 men who were diagnosed with liver steatosis using ultrasound/liver biopsy. We examined 75 men, who were diagnosed for having liver steatosis (ultrasound/liver biopsy). Each filled in a questionnaire on alcohol intake. Subjects were divided into two subgroups according to alcohol history and CDT concentrations- drinkers and non-drinkers. All individuals were examined for serum concentrations of AN, glucose, triglycerides, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and glutamate tranferase (GMT) activity; carbohydrate-deficient transferrin (CDT %) a marker of chronic alcohol consumption, insulin and TNF-α. The Quicki insulin sensitivity index was calculated. Forty-eight individuals were found to be moderate drinkers and 27 subjects non-drinkers. Moderate drinkers had significantly higher concentrations of AN (13.8 ± 3,7 versus 9.1 ± 5.4 mg/l, means ± SD, p = 0.012) compared with non-drinkers, independent of adiposity. Plasma AN concentrations in the whole group were positively correlated with TNF-α concentrations (r = 0.6; p = 0.0001), CDT (r = 0.26; p = 0.0084), AST/ALT index (r = 0.3, p = 0.009), AST (r = 0.29; p = 0.011) and GMT (r = 0.29; p = 0.011) and negatively with BMI (r = – 0.48; p = 0.0002) and glycemia (r = – 0.22; p = 0.049). The positive associations of AN with TNF-α (0.8; p = 0.001), CDT (0.55; p = 0.017), AST/ALT index (0.55; p = 0.019) and the negative correlation with glycemia (– 0.35; p = 0.0158) were independent of BMI. Stratified according to alcohol intake, in moderate drinkers, a positive correlation was found between AN and TNF-α concentrations (r = 0.6, p = 0.0001, AST/ALT index (r = 0.34, p = 0.0295) whereas in non-drinkers no such correlations were found. The concentration of AN and BMI displayed a negative correlation in both drinker and nondrinker patients (r = – 0.42, p = 0.01 and – 0.61; p = 0.012, respectively). We concluded that plasma AN is higher in moderate drinkers compared to non-drinkers, even after correction for BMI. Drinkers suffering from liver steatosis were found to have a positive correlation between AN concentrations, laboratory markers of liver disease and TNF-α. Such correlation was absent in non-drinkers suffering from liver steatosis. This suggests that alcohol may modulate the inhibitory effect of TNF-α on AN production, and thus, increase its plasma concentrations.

Microtubule disruptors and their interaction with biotransformation enzymes.

Abstract: Microtubule disruptors, widely known as antimitotics, have broad applications in human medicine, especially as anti-neoplastic agents. They are subject to biotransformation within human body frequently involving cytochromes P450. Therefore antimitotics are potential culprits of drug-drug interactions on the level of activity as well as expression of cytochromes P450. This review discusses the effects of four well-known natural antimitotics: colchicine, taxol (paclitaxel), vincristine, and vinblastine, and a synthetic microtubule disruptor nocodazole on transcriptional activity of glucocorticoid and aryl hydrocarbon receptors. It appears that microtubules disarray restricts the signaling by these two nuclear receptors regardless of cell cycle phase. Consequently, intact microtubules play an important role in the regulation of expression of cytochromes P450, which are under direct or indirect control of the two nuclear receptors.

Phenytoin induced oxidative stress in pre- and postnatal rat development - effect of vitamin E on selective biochemical variables.

Abstract: A pre- and postnatal study was carried out to investigate the effect of high dose (500 mg/kg) of the natural antioxidant vitamin E (VIT E) on biochemical variables in the model of chronic intrauterine hypoxia. Chronic hypoxia was induced by administration of the anticonvulsant phenytoin (PHT) during pregnancy. Rats were orally treated with PHT (150 mg/kg) from day 7 to 18 of gestation and VIT E prior to PHT orally on the same days. The activity of the lysosomal enzyme N-acetyl-ss-D-glucosaminidase (NAGA) and the level of glutathione (GSH) were used as markers of tissue damage. In the prenatal study PHT-induced embryofoetal toxicity was associated with an increase in NAGA activity and decrease of GSH level in maternal serum and heart and with an increase in NAGA activity in the placenta. Administration of VIT E did not inhibit the above given changes. PHT increased the activity of NAGA and decreased the level of GSH in foetal organs (liver, lungs, brain). VIT E did not reverse these changes. In the postnatal study, we did not find any significant differences in NAGA activity in the organs of 1-day-old pups. An increase of liver GSH level was found in PHT and VIT E+PHT groups of pups and in the group VIT E+PHT in the lungs. In conclusion, supplementation with a high-dose of VIT E failed to protect maternal, foetal and new-born rat organs from PHT induced changes of selective biochemical variables.

The role of natural biopolymers in genotoxicity of mutagens/carcinogens elimination.

Abstract: Nowadays naturally occurring compounds with the potential antimutagenic and anticarcinogenic effects are of great importance for their prospective use in cancer chemoprevention and treatment. The new water soluble derivative of microbial polysaccharide beta-D-glucan-carboxymethyl glucan (CMG) belongs to such a category of natural substances. CMG isolated from the cell wall of baker's yeast Saccharomyces cerevisiae is included into the class of biopolymers known as biological response modifiers (BRMs) with a broad range of activities, above all ones interfering with cancer therapy. It was demonstrated on four experimental model systems that biological and consequential medicinal importance of CMG is based on the combined application with another active compound. In the Saccharomyces cerevisiae antimutagenicity assay CMG significantly reduced ofloxacin-induced mutagenicity in the yeast strain D7. CMG exerted bioprotective (anti-toxic and antimutagenic) effect after its simultaneos application with methyl methanesulphonate on the repair-deficient strain uvs10 of the unicellular green alga Chlamydomonas reinhardtii. In the Vicia sativa simultaneous phytotoxicity and anticlastogenicity assay CMG exerted statistically significant anticlastogenic efect against maleic hydrazide-induced clastogenicity in Vicia sativa L. Only in the Salmonella/microsome assay CMG did not exert statistically significant antigenotoxic effect, despite of the fact that it reduced 9-aminoacridine-induced mutagenicity in S. typhimurium TA97, but his(+) revertants decreasing was statistically significant only at the highest CMG concentration used. The data presented unambiguously documented that even biopolysaccharides (e.g., derivatives of beta-glucan) belonging to the most abundant class of natural biopolymers may contribute to cancer prevention and therapy.

What are artificial neural networks and what they can do

Abstract: The artificial neural networks (ANN) are very often applied in many areas of toxicology for the solving of complex problems, such as the prediction of chemical compound properties and quantitative structure-activity relationship. The aim of this contribution is to give the basic knowledge about conception of ANN, theirs division and finally, the typical application of ANN will be discussed. Due to the diversity of architectures and adaptation algorithms, the ANNs are used in the broad spectrum of applications from the environmental processes modeling, through the optimization to quantitative structure-activity relationship (QSAR) methods. In addition, especially ANNs with Kohonen learning are very effective classification tool. The ANNs are mostly applied in cases, where the commonly used methods does not work.

Effect of long-term administration of arsenic (III) and bromine with and without selenium and iodine supplementation on the element level in the thyroid of rat.

Abstract: The aim of this study was to evaluate the influence of arsenic and bromine exposure with or without iodine and selenium supplementation on the element level in the thyroid of rats. Four major groups of Wistar female rats were fed with respective diets: group A - standard diet, group B - iodine rich diet (10 mg I/kg food), group C - selenium rich diet (1 mg Se/kg) and group D - iodine and selenium rich diet (as in group B and C). Each group was divided into four subgroups per 7 animals each receiving either NaAsO(2) ip (6.5 mg.kg(-1) twice a week for two weeks and 3.25 mg.kg(-1) for six weeks) or KBr in drinking water (58.8 mg.l(-1)) for 8 weeks or combined administration of both substances. Remaining subgroup served as controls. After 8 weeks thyroid glands were analyzed by ICP-MS for As, Br, Se, and I content. The exposition of rat to arsenic or bromine causes the accumulation of these elements in the thyroid gland ( approximately 18 ppm of As, approximately 90 ppm of Br) and significantly affects iodine and selenium concentration in the thyroid. In iodine and/or selenium supplemented rats the bromine intake into the thyroid was lowered to approximately 50% of the level in unsupplemented animals. Also selenium thyroid level elevated due to KBr administration was lowered by iodine supplementation in the diet. The accumulation of arsenic in the thyroid was not influenced by selenium or iodine supplementation; however, As(III) administration increased iodine thyroid level and suppressed selenium thyroid level in selenium or iodine supplemented group of animals.

Results of selfmonitoring on glucometer systems Advance and Optium in daily routine.

Abstract: The aim of this prospective clinical study was to compare the results of B-glucose estimations performed simul-taneously on glucometer Advance (with Micro-draw strips) and Optium (G3 strips) by lay healthy volunteers under non-standardized conditions of everyday life, to assess the difficulties dealing with lay – handling of these systems and to demonstrate the possibilities of the software Glucobalance (Hypoguard) and PC-Link (Medisense/Abbott) for the analysis of selfmonitoring. In the course of 5 days, a total of 721 pairs of measurements were carried out on 10 pairs of glucometer Advance and Optium by 10 healthy volunteers aged 16–40 years. The data transfer of all values into computer from glucometer Advance using the Glucobalance software and from glucometer Optium using the PC-Link was carried out to determine the results. The correlation of B-glucose measured on the glucometer Advance and Optium was strong (r = 0.73). Glucometer Advance brings values about 0.21 ± 0.06 mmol/l lower than gluco-meter Optium. The average difference found within each pairs of glucometers Advance – Optium varied. Nevertheless, these differences are acceptable for routine selfmonitoring. The handling of glucometer Advance is not difficult for lay persons. The Glucobalance software simplifies the result evaluation by each tested person. Even though there are some advantages in comparison with the PC-Link, it should be further developed.INTRODUCTIONBlood glucose selfmonitoring has become a prerequi-site in the successful treatment of both type 1 and type 2 diabetes mellitus

Cytotoxicity of pivoxil esters of antiviral acyclic nucleoside phosphonates: adefovir dipivoxil versus adefovir.

Abstract: Biological effectiveness of antiviral acyclic nucleoside phosphonate adefo vir, 9-[2-(phosphonomethoxy)ethy]ade nine (PMEA) and its more lipophilic (bis)pivaloyloxymethyl ester prodrug adefovir dipivoxil (bis-POM-PMEA) were compared under in vitro conditions in mammalian cell systems. Proliferation of murine splenocytes was inhibited in a concentration-dependent manner, the bis-POM-PMEA being more effective than PMEA. In contrast to PMEA, bis-POM-PMEA inhibited production of nitric oxide (NO) in macrophages activated with interferon-γ (IFN-γ) and lipopolysaccharide (LPS). Viability of both splenocytes and macrophages remained uninfluenced by PMEA, whereas pronounced cytocidal effects were exhibited by bis-POM-PMEA. The IC 50 s reached the values of 15 µM and 30 µM in cultures of macrophages and splenocytes, respectively (assayed at the interval of 24 hrs). The effects could partly be mimicked by formaldehyde, a decomposition product of the pivoxil moiety of bis-POM-PMEA. The other possible product, pivalic acid, was ineffective in this respect. The present data are consistent with the view that pivoxil prodrug of PMEA, bis-POM-PMEA possesses enhanced but also broader spectrum of biological effects than the parent compound.