Cytoprotective effect of silybin against lasalocid-induced toxicity in HepG2 cells.

Lasalocid is an ionophore coccidiostatic agent frequently used in poultry. Its extensive use causes the formation of residues in edible tissues and eggs which may pose a risk to consumers. Silybin is the main compound extracted from the herb milk thistle Silybum marianum and its hepatoprotective effect has been reported in literature. The aim of the study was to compare lasalocid and silybin cytotoxic effects followed by their combined use in HepG2 cell line. A cytoprotective effect resulting from the interaction of both pharmacologically active substances was measured. In this study, an MTT test, coomassie brillant blue binding test, and LDH release test determined the effective concentration (EC50) of the compounds. The isobolograms and combination index were used to assess the nature of interaction. The lowest EC50-value for lasalocid was established via the MTT test. This study revealed a lack of silybin cytotoxic effect on the cells. Co-actions of the two drugs led to a significant decrease of lasalocid cytotoxicity. The isobolograms and combination index showed a remarkable antagonistic effect in the course of silybin and lasalocid interaction. The results indicate that silybin revealed a cytoprotective effect when incubated with lasalocid since its cytotoxic impact on HepG2 cells has been significantly diminished.

Circulating endothelial progenitor cells are inversely correlated with in-stent restenosis in patients with non-ST-segment elevation acute coronary syndromes treated with EPC-capture stents (JACK-EPC trial).

AIM: Aim of the study was to evaluate the association between circulating endothelial progenitor cells (EPCs) and angiographic outcomes after implantation of GenousTM stent in patients with non-ST-segment elevation acute coronary syndromes (ACS) (NSTE-ACS) undergoing urgent percutaneous coronary intervention (PCI). METHODS: Sixty patients treated with EPC-capture stent (N.=30) or bare metal stents (BMS) (N.=30) receiving 80 mg atorvastatin and dual antiplatelet therapy (DAT) for 12 months. Restenosis was assessed after 6 months by quantitative coronary angiography (QCA) and major acute coronary events (MACE) evaluated after 6 and 12 months. INCLUSION CRITERIA: de novo lesion >70% in native vessel, diameter 2.5-4 mm, lesion length <30 mm. EXCLUSION CRITERIA: diabetes, previous revascularization, significant left main stenosis, chronic total occlusions (CTO) and multivessel disease. RESULTS: Majority of patients in EPC-capture stent and BMS groups presented with NSTEMI (73.3% and 70%, respectively). Mean stent length was 20.1±8 and 19.9±10 mm, diameter 3±0.97 and 3.1±0.88 mm in respective groups. The binary restenosis was significantly lower in GenousTM (13 vs. 26.6%, P=0.04). Risk of MACE after 6 and 12 months were comparable in both groups. There was no stent thrombosis. Numbers of circulating EPCs were significantly approximately 2-fold higher during the ACS than after 6 months. Mobilization of EPCs during acute ischemia was significantly lower in patients who developed restenosis after 6 months (3 vs. 4.5 cells/µL, P=0.002) and it was negatively correlated with late-loss after 6 months (R=-0.42; P<0.03). CONCLUSION: Use of GenousTM stents in NSTE-ACS is associated with lower restenosis rate than BMS at 6 months. There was no ST through 1 year. The number of circulating EPCs is inversely correlated with in-stent late loss (LL).

Contents of zinc, copper, chromium and manganese in silver foxes according to their age and mineral supplementation.

Serum, livers and kidneys of 30 silver foxes from one breeding farm were subjected to analysis of the four microelements contents. The samples derived from 3 groups of animals (n = 10) selected according to age and developed reproduction disorders. Cu, Cr and Mn were determined by a graphite furnace AAS whereas Zn by the flame AAS methods. Serum levels of Zn were the least variable (x (n = 30) = 4.72 +/- 2.313 microg x mL(-1)) and the mean of Cu content was 0.26 +/- 0.244, of Cr was 0.029 +/- 0.032 and of Mn was 0.074 +/- 0.085 microg x mL(-1). The livers and kidneys contained respectively: 159.9 +/- 23.66 and 74.25 +/- 14.44 microg g(-1) of Zn; 34.03 +/- 12.43 and 13.66 +/- 1.67 microg g(-1) of Cu as well as 6.28 +/- 0.97 and 2.60 +/- 0.33 microg g(-1) of Mn. The most variable was Cr level achieving 1.00 +/- 1.06 and 1.43 +/- 2.64 microg g(-1) (all results per gram of wet weights). The differences between means and medians within the age groups did not exceed 41%, however chromium was an exception, its values differed more than 2 times. High zinc level found both in the serum and the organs resulted from its concentration in feedstuff being 1040,5 microg g(-1), exceeding the recommended level for farming foxes. Over-supplementation of dietary Zn might suggest connection between subtoxic action of Zn or its interaction with Cu or Cr followed by subsequent effect on reproduction events. Whether nature of Zn effects derived from direct target action or from Zn - Cu or Zn - Cr interactions have to be solved experimentally. Some foxes contained copper and chromium in livers and kidneys at levels exceeded significantly those concidered as physiological ranges for mammals, whereas manganese was within these limits. Thus, manganese was considered to keep homeostasis status of this element in the examined foxes.

Binding of 3H-metronidazole in olfactory, respiratory and alimentary epithelia in rats.

Whole-body autoradiography of 3H-metronidazole in rats showed retention of bound metabolites in the epithelia lining the olfactory part of the nose, the tongue, the gingiva, the palate, the pharynx, the oesophagus and the forestomach. In vitro microautoradiography in O2- and N2-atmosphere with some of these tissues indicated reductive formation of bound metabolites in specific cells of the epithelia. Studies with subcellular fractions of the nasal olfactory mucosa showed formation of DNA- and protein-bound metronidazole metabolites. A lower bioactivating capacity was found in experiments with the liver. The bioactivation was dependent on N2-atmosphere, and presence of the P450-inhibitor metyrapone or GSH in the incubation media depressed the protein-binding of metronidazole both in the nasal olfactory mucosa and the liver. These data indicate that the bioactivation is partly P450-dependent and GSH may play an important role in scavaging the bioactivated drug. The epithelial cells with a capacity to bioactivate metronidazole may be potential targets for negative effects of the drug. Whole-body autoradiography also showed a strong binding of radioactivity in the contents of caecum and colon. This can be considered to be due to reductive bioactivation of metronidazole by the intestinal microorganisms and reflects the principal site of action of the drug.

Disposition of metronidazole in hens (Gallus gallus) and quails (Coturnix coturnix japonica): pharmacokinetics and whole-body autoradiography.

Hens were given single intravenous or oral doses (30 mg/kg body weight) of metronidazole and the plasma concentrations of the drug were determined by high-performance liquid chromatography (HPLC) at intervals from 10 min to 24 h after drug administration. Pharmacokinetic variables were calculated by the Lagrange algorithm technique. The elimination half-life (t1/2 beta) after the intravenous injection was 4.2 +/- 0.5 h, the volume of distribution (Vd(ss) 1.1 +/- 0.2 L/kg and the total body clearance (ClB) 131.2 +/- 20 mL/h.kg. Oral bioavailability of the metronidazole was 78 +/- 16%. The plasma maximum concentration (Cmax) 31.9 +/- 2.3 micrograms/mL was reached 2 h after the oral administration and the oral elimination half-life (t1/2 beta) was 4.7 +/- 0.2 h. The binding of metronidazole to proteins in hen plasma was very low (less than 3%). Whole body autoradiography of [3H] metronidazole in hens and quails showed an even distribution of labelled material in various tissues at short survival intervals (1-4 h) after oral or intravenous administration. A high labelling was seen in the contents of the small and large intestines. In the laying quails a labelling was also seen in the albumen and in a ring in the periphery of the yolk at long survival intervals. Our results show that a concentration twofold above the MIC is maintained in the plasma of hens for at least 12 h at an oral dose of 30 mg/kg metronidazole.

[Lead, cadmium, copper and zinc content in vegetables, gooseberry fruit and soil from gardening plots of Lublin]. / Zawartosc olowiu, kadmu, miedzi i cynku w warzywach, owocach agrestu oraz glebie ogrodów dzialkowych Lublina.

Lead, cadmium, copper and zinc contents in vegetables, fruits of gooseberries and in soil of Lublin gardening plots "Pionier" and "Podzamcze" both situated along heavy traffic streets, and "Pionier" additional to the close vicinity of automobile factory were determined by atomic absorption spectrophofometry. Statistical analysis of the results respected the streets or factory distances from the gardening plots, and the species of the vegetables. Levels of the determined elements in the most of the samples were lower than permitted by the Ministry of Health Regulation established in 1993. According to that Regulation a slight exceeding of zinc content in the red beetroots and in the overground parts of the leak were found. Lead content in the parsley root and in the dill was close to the tolerance limit. Both lead and cadmium of the soil of "Podzamcze" gardening plot significantly exceeded the levels considered as tolerable for unpolluted grounds, whereas the soil of "Pionier" contained lead close to limit of tolerance level.

Prochymosin and pepsinogen potential activity and electrophoretic study of proteolytic fractions in the gastric and duodenal mucosal extracts from the first four day old lambs.

Potential proteolytic activity of prochymosin (PCh) and pepsinogen (PG) as well as relative activity of the proteolytic fractions in the mucosal extracts of the whole stomach, pyloric region of the stomach and duodenum were investigated. The main activity in the lambs stomach after birth demonstrated PCh however a small amount of PGA activity was also present. The highest activity of PCh from the whole stomach was observed in day 2 and 3, PG activity from the whole and pyloric region of the stomach increased in the first day of life and later was not significant by changed. Mucosal extracts of the whole stomach contained 3 to 4 of pepsinogen fractions of the fast migration (Pg1, Pg2, Pg3 and Pg4) in which Pg3 was of the highest activity in the majority of lambs. Pyloric region revealed 3 fractions: Pg2, of a small and Pg3 and Pg4 of high and equal activity. In duodenum Pg3 and Pg4 were of equal activity. The beginning of Pg2, Pg3 and Pg4 relative activity changes were observed in lambs stomach in respect to increase of Pg2 and decrease of Pg3 activity. The group of 3-4 fractions of slow electrophoretic migration contained probably progastricsin of small--and prochymosin of the highest activity within this group and a small non-pepsinogen fraction.

Immunohistochemical studies on the development of cells containing progastricsin (minor pepsinogen) in comparison to prochymosin and pepsinogen in bovine abomasal mucosa.

The localization of progastricsin was studied in cells of abomasal mucosa from cattle of different ages and feeding regimes and compared to the localization of prochymosin and pepsinogen in the same material by use of an immunofluorescence technique with specific rabbit antibodies. Immunoreactivity for progastricsin was first found in calves at the age of about 45 days in surface mucous cells in the pit of the fundic gland. In older calves and adults, mucous neck cells also produced progastricsin. In the pyloric mucosa, on the other hand, traces of progastricsin immunoreactivity were found in the lower base of the pyloric gland even in newborn calves. When the calves grew older, progastricsin-immunoreactive cells also developed in the pit and later in the neck of the pyloric gland; and the number of these cells in this region increased with age. The development of progastricsin-producing cells seemed to be influenced only by age and not by the feeding of milk to the calves. The ontogeny of progastricsin, prochymosin, and pepsinogen exhibited an interesting pattern in cattle, as they started to be produced at three different ages and gave three different patterns of development in the cells of abomasal mucosa. The number of cells producing prochymosin was closely correlated with milk-feeding, while the development of progastricsin was most related to the age of the calf. The most stable factor during the development of the cells in the abomasum was the number of cells producing pepsinogen.

Effect of chloramphenicol on the activity of alkaline phosphatase (EC 3.1.3.1.) and the structure of microvilli of the epithelial cells of the small intestine of the hen.

Effect of a therapeutic dose of chloramphenicol (detreomycin) on the intensity of reaction of alkaline phosphatase and on the structure of microvilli of the epithelial cells of the small intestine of the hen was investigated. It was observed that this antibiotic given orally weakened the intensity of the reaction of alkaline phosphatase and changes also the shape of microvilli of intestinal epithelium. 4 weeks after chloramphenicol administration the activity of the enzyme appeared to be normal and the morphology restored. We concluded from the experiments performed, that chloraphenicol may inhibit the process of protein glycosylation in the enterocytes of the small intestine and also weakened reversible the process of absorption.