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1

Ograniczanie wchlaniania trucizn z przewodu pokarmowego u psow i kotow

100%
Bakala A., Wiechetek M.
Życie Weterynaryjne
|
2003
|
tom 78
|
nr 08
450-454
2

Hypothermic storage of equine isolated hepatocytes

80%
Bakala A., Karlik W., Wiechetek M.
Polish Journal of Veterinary Sciences
|
2007
|
tom 10
|
nr 1
The aim of the study was to establish the optimal methods for hypothermic storage of equine isolated hepatocytes. Viability of equine isolated hepatocytes after hypothermic storage was dependent on the type of storage medium as well as on the cell density in the storage suspension and the preservation period. Hepatocytes stored at 4°C in Hanks' Balanced Salt Solution (HBSS) and Williams' Medium E (WE) for 24 h showed very low viability, numerous cell membrane blebs, very low attachment rate (11.9 ± 6.5% and 34.8 ± 19.1%, respectively) and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) reduction rate (6.4 ± 3.9% and 25.1 ± 14.8%, respectively). In contrast, hepatocytes stored in University of Wisconsin Solution (UW) after 24 h of storage at a density of 12.5 x 106 cells/ml showed high viability (over 70%), typical and intact morphology, high cell attachment rates and MTT reduction. Our findings clearly demonstrate that UW is a good preservation solution for equine isolated hepatocytes. Hepatocytes harvested from slaughterhouse organs can be stored at 4°C in UW at a density of 12.5 x 106 cells/ml for at least 24 h without significant decrease in functional integrity.
3

Optimization of the technique for the short-term culture of equine hepatocytes

71%
Bakala A., Karlik W., Wiechetek M., Grono D., Glowala A.
Medycyna Weterynaryjna
|
2007
|
tom 63
|
nr 11 Supl.
1440-1442
The aim of this study was to establish the optimal conditions for the culture of equine hepatocytes in a monolayer configuration. The obtained results show that the rate of MTT metabolism correlated with the number of cultured cells and a linear increase of MTT reduction rate was observed in cases when the cell density varied between 1.25 × 10⁴ to 5 × 10⁴ viable cell/well of 96-well plate. Hepatocytes reached the optimal cell attachment rate and MTT reduction at a cell density of 5 × 10⁴ cells/well. The number of attached cells to a plastic culture dish was also related to incubation time. The greatest ability of hepatocytes to attach to the culture dish was observed after 10 h of incubation and it was found to be 84.1 ± 2.5% of seeded hepatocytes. It was also found that fetal bovine serum was more efficient than horse serum for the attachment of equine isolated hepatocytes in a monolayer culture. The highest rate of cell attachment (assessed microscopically and with MTT reduction test) was observed when cells were plated with the culture medium supplemented with FBS or HS at a concentration of 5%. However, medium supplementation with higher than 5% serum concentration (10% of FBS or HS) significantly decreased MTT reduction rate. The rate of MTT metabolism and cell attachment in hepatocytes cultured in WE supplemented with FBS or HS was also dependent on the plating time and were the highest after 10 h of seeding.
4

Aktywność trzeciej rodziny Cytochromu P450 w izolowanych hepatocytach konia

71%
Grono D., Glowala A., Bakala A., Karlik W., Wiechetek M.
Prace i Materiały Zootechniczne
|
2008
|
tom 66
5

Papaverine should not be used as an indicator for controlling the ability of the gastrointestinal tract muscle to relax in vitro

71%
Chlopecka M., Dziekan N., Bakala A., Wiechetek M., Mendel M.
Medycyna Weterynaryjna
|
2007
|
tom 63
|
nr 11 Supl.
1437-1439
The aim of this research was to study the possibilities of using papaverine as a reference substance to control the ability of isolated gastrointestinal (GI) tract strips to relax. The effects of papaverine hydrochloride (0.001-100 µM) dissolved in distilled water, or in DMSO (0.5%), on the mechanical activity of isolated rat GI strips (stomach fundus and corpus, duodenum and jejunum) were studied. The obtained results show that papaverine provoked various responses of the examined muscle strips dependent on the part of GI tract and papaverine solvents used (water or DMSO). Papaverine applied as water solution caused muscle relaxation of all investigated gastrointestinal strips: the lowest effective (induced relaxation) concentration of papaverine was 10 µM for gastric corpus, jejunum and duodenum and 100 µM for gastric fundus. However, there was no dependence between the concentration of papaverine and the degree of muscle relaxation of the studied GI strips. Moreover, in case of gastric fundus strips, papaverine applied as 0.5% DMSO solution provoked different muscle responses: in the presence of 0.1 and 1 µM papaverine contraction occurred; administering papaverine at higher concentrations (10 and 100 µM) resulted in relaxation. The obtained results clearly indicate that papaverine does not fulfil the criteria set for the reference substance and should not be used as an indicator for controlling of gastrointestinal tract muscle relaxation in vitro.
6

Porównanie kinetyki przemian kumaryny w modelu perfundowanej wątroby i zawiesinie hepatocytów konia

71%
Karlik W., Bakala A., Glowala A., Grono D., Wiechetek M.
Prace i Materiały Zootechniczne
|
2008
|
tom 66
7

Skład medium inkubacyjnego a aktywność metaboliczna izolowanych hepatocytów konia w długoterminowej hodowli

71%
Glowala A., Grono D., Bakala A., Karlik W., Wiechetek M.
Prace i Materiały Zootechniczne
|
2008
|
tom 66
8

Porownanie aktywnosci metabolicznej bioreaktora watrobowego z perfundowana watroba

61%
Karlik W., Sztuk P., Bakala A., Glowala A., Grono D., Wiechetek M.
Medycyna Weterynaryjna
|
2008
|
tom 64
|
nr 03
354-358
The aim this study was to compare metabolic activity of a hollow fiber bioreactor with a perfused liver. A special construction of a hollow fiber bioreactor was used with freshly isolated rat hepatocytes. After isolation, hepatocytes were incubated in the bioreactor for the duration of 3 hrs in the following conditions: 100 ml medium Hanks Balances Salts Solution supplemented with 4% albumin, 2 mM ornithine, 10 mM ammonium chloride and 6 mM ethanol were used; the medium was perfused in a circulated system for 25 ml/min; samples of the medium were taken to estimate ammonia, urea, glucose, lactate, ethanol, AST and ALT activity before and after 5 min and every 15 min of perfusion time. The same experimental condition was used in the perfused rat liver system. Utilization of ammonia was different in both systems and amounted to: 8.89 and 5.23 µmol/h/g hepatocytes in the bioreactor and perfused liver, respectively. Urea production was: 2.35 and 8.22 µmol/h/g hepatocytes, respectively. The largest differences between the compared systems were observed in the glucose and lactate metabolism. The bioreactor did not release glucose and lactate during the entire time of perfusion. In contrast, perfused liver intensively released glucose (31.32 µmol/hr/g cells) and produced lactate (29.42 µmol/hr/g cells). On the other hand, there was no statistically significance difference in the rate of ethanol metabolism between both systems, which amounted to 2.55 and 2.04 umol/h/g hepatocytes in the bioreactor and perfused liver, respectively. The results indicate that a bioreactor with freshly isolated hepatocytes is not bioequivalent to a perfused liver if estimation is made on the basis of ureogenesis and/or glucose utilization. However, ethanol utilization gives evidence that the metabolic activity of the bioreactor is comparable with a perfused liver. On the basis of the obtained results it can be concluded that the difference in metabolic activity of the bioreactor and perfused liver is connected with catabolic state and disturbances of energy metabolism in freshly isolated hepatocytes. In such a condition HBSS is not the proper medium for the recovery of homeostasis. To estimate the metabolic activity of freshly isolated hepatocytes cultivated in various in vitro systems such as a marker of model usefulness it is suggested to use the activity of inducible enzymes, but not constituent enzymes.
9
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Evaluation of the time-stability of an alternative research model based on isolated rat gastrointestinal strips

61%
Chlopecka M., Dziekan N., Mendel M., Bakala A., Maldyk J., Wiechetek M.
Journal of Physiology and Pharmacology. Supplement
|
2007
|
tom 58
|
nr 3
73-86
10

Wplyw kadmu na aktywnosc enzymow antyoksydacyjnych w izolowanych hepatocytach szczura

51%
Czeczot H., Scibior-Bentkowska D., Skrzycki M., Podsiad M., Karlik W., Bakala A., Grono D., Wiechetek M.
Medycyna Weterynaryjna
|
2009
|
tom 65
|
nr 01
55-60
The aim of the study was to evaluate the effect of cadmium on the antioxidant enzyme activity in rat hepatocytes. The experiments were conducted on isolated rat hepatocytes, which were incubated for two hours in modified Williams’ E medium (MWE) with 25, 50, and 200 µM cadmium chloride (CdCl₂). Hepatocytes incubated in MWE medium without cadmium chloride were used as a control. The activity of antioxidant enzymes - superoxide dismutases (SOD1 and SOD2), catalase (CAT), total glutathione peroxidase (tot. GSHPx), selenium-dependent glutathione peroxidase (Se-GSHPx), glutathione S-transferase (GST) and glutathione reductase (GSHR) - and the values of the studied oxidative stress markers - the concentration of tiobarbituric-acid-reacting substances (TBARS) and reduced glutathione (GSH) - indicate that cadmium induces oxidative stress in rat hepatocytes, which impairs antioxidative mechanisms.
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