This study was based on our previously developed double-layered enterohepatic co-culture system, ... more This study was based on our previously developed double-layered enterohepatic co-culture system, composed of nontumorigenic porcine intestinal epithelial cell line (IPEC-J2) and primary culture of porcine hepatocytes. The anti-inflammatory effect of spent culture supernatant of Lactobacillus plantarum 2142 (Lp2142; 13.3%) and sodium n-butyrate (2 mM) was tested on IPEC-J2 and hepatocyte monocultures as well as on the gut-liver co-culture. To mimic inflammation, lipopolysaccharide (LPS; 1 and 10 μg/mL) was applied. Production of IL-8 and IL-6 was measured as a marker of inflammatory responses. The paracellular permeability of the intestinal epithelium was also monitored by fluoresceinisothiocyanate-labeled dextran 4 assay. Significant increase of IL-8 concentration was observed in the IPEC-J2 monoculture (P < 0.01) while the level of IL-6 was not changed following LPS treatment. Concentration of IL-8 and IL-6 was grown significantly in hepatocyte monocultures (P < 0.05 and P < 0.001) as well as in the co-culture after 10 μg/mL LPS treatment (P < 0.001 and P < 0.001). One microgram per milliliter LPS caused elevated IL-8 level in the co-culture (P < 0.001) and in the hepatocyte monoculture (P < 0.01), while it caused increased IL-6 level only in the hepatocytes (P < 0.001). Production of IL-8 was significantly decreased by butyrate in case of 1 μg/mL as well as 10 μg/mL LPS exposure in the co-culture (P < 0.001). Application of butyrate also reduced IL-6 level in the co-culture after 10 μg/ mL LPS treatment (P < 0.01). Lactobacillus plantarum 2142 decreased IL-8 level after incubation with 1 μg/ mL LPS (P < 0.001), while in case of 10 μg/mL LPS treatment only a marginal lowering in IL-8 (P = 0.064) release was measured. The IL-6 concentration was significantly reduced (P < 0.01 in case of 1 μg/mL LPS treatment) by Lp2142 in the co-culture. Contrarily, the elevated IL-8 and IL-6 level of hepatocytes has not been reduced in case of either butyrate or Lp2142 addition. The enterohepatic co-culture model offers a possibility for fast and reliable screening of new candidates against enteric inflammation, which are of special interest in porcine medicine and health management. According to our results, Lp2142 and butyrate both seem to be effective as anti-inflammatory agents in LPS-triggered inflammatory response, tested in the gut-liver co-culture model.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
The authors investigated the possibility of the presence of VTEC strains in improperly pasteurize... more The authors investigated the possibility of the presence of VTEC strains in improperly pasteurized milk samples. A total of 64 Escherichia coli strains were isolated from 135 pasteurized milk samples originating from the same producer. The examined isolates contained 29 haemolysin-, 9 colicin-and 5 aerobactin-producing strains, but the investigations concerning heat-resistant and heat-sensitive toxins gave negative results. Six O128-type E. coli strains exerted a cytotoxic effect on the VERO cell line; 5 of them contained H12 antigen, while one could not be typed. Four of the 6 verocytotoxin-producing strains belonged in phage group 20, one in phage group (2)3(7), and one in phage group 4; four strains were of B3, one of A1, and one of A1(A2) phage type. Because of a technical failure the milk was pasteurized at 69 ºC for 15 s, which is 2 ºC less than required. The results underline the importance of the appropriate pasteurization temperature, as otherwise the milk may contain verocytotoxin-producing E. coli, which is a potentially great hazard for public health.
Cytochrome P450 (CYP) oxidases are among the main metabolizing enzymes that are responsible for t... more Cytochrome P450 (CYP) oxidases are among the main metabolizing enzymes that are responsible for the transformation of xenobiotics, including clinically important drugs. Their activity can be influenced by several compounds leading to decreased efficacy or increased toxicity of co-administered medicines. Flavonoids exert various beneficial effects on human and animal health; therefore they are used as food and feed supplements. However, they are also well-known for their CYP modulating potential. Since the amount of CYP enzymes is highest in the liver, interaction studies are mainly conducted in hepatocytes, however, CYP activity in the gastrointestinal tract is also remarkable. In this study, effects of apigenin (API), quercetin (QUE) and their methylated derivatives trimethylapigenin (TM-API), 3-O-methylquercetin (3M-QUE) and 3′,7-di-O-methylquercetin (3′7DM-QUE) on the CYP enzyme activity was examined in IPEC-J2 porcine intestinal epithelial cells. Potential food-drug interactions...
Journal of Veterinary Pharmacology and Therapeutics, 2014
Butyrate, a commonly applied feed additive in poultry nutrition, can modify the expression of cer... more Butyrate, a commonly applied feed additive in poultry nutrition, can modify the expression of certain genes, including those encoding cytochrome P450 (CYP) enzymes. In comparative in vitro and in vivo experiments, the effect of butyrate on hepatic CYP genes was examined in primary cultures of chicken hepatocytes and in liver samples of chickens collected from animals that had been given butyrate as a feed additive. Moreover, the effect of butyrate on the biotransformation of erythromycin, a marker substance for the activity of enzymes of the CYP3A family, was investigated in vitro and in vivo. Butyrate increased the expression of the avian-specific CYP2H1 both in vitro and in vivo. In contrast, the avian CYP3A37 expression was decreased in hepatocytes following butyrate exposure, but not in the in vivo model. CYP1A was suppressed by butyrate in the in vitro experiments, and overexpressed in vivo in butyrate-fed animals. The concomitant incubation of hepatocytes with butyrate and erythromycin led to an increased CYP2H1 expression and a less pronounced inhibition of CYP3A37. In in vivo pharmacokinetic experiments, butyrate-fed animals given a single i.m. injection of erythromycin, a slower absorption phase (longer T(half-abs) and delayed T(max)) but a rapid elimination phase of this marker substrate was observed. Although these measurable differences were detected in the pharmacokinetics of erythromycin, it is unlikely that a concomitant application of sodium butyrate with erythromycin or other CYP substrates will cause clinically significant feed-drug interaction in chickens.
Effects of orally applied butyrate bolus on histone me P450 enzyme activity Conclusion: Orally ad... more Effects of orally applied butyrate bolus on histone me P450 enzyme activity Conclusion: Orally added butyrate in bolus could cause in vivo hyperacetylation of the hepatic core histones, Mátis et al. Nutrition & Metabolism 2013, 10:12
Az akut oxidativ stressz es a probiotikumok hatasanak vizsgalatahoz kollagennel bevont polieszter... more Az akut oxidativ stressz es a probiotikumok hatasanak vizsgalatahoz kollagennel bevont polieszter membran inzerten tenyesztett, ujszulott sertes jejunum eredetű IPEC-J2 sejtvonalat alkalmaztunk. A peroxiddal kezelt IPEC-J2 sejtekben a gyulladasos citokinek (IL-8 es TNF-?) es a vedő hatasu hősokk feherje 70 (Hsp 70) relativ genexpressziojat kvantitativ real-time PCR-rel hataroztuk meg a probiotikus bakteriumtorzsek (Lactobacillus plantarum 2142, Lactobacillus casei Shirota, Bifidobacterium animalis subsp. lactis BB-12, Bacillus amyloliquefaciens CECT 5940, Enterococcus faecium CECT 4515) feluluszojanak (SMSz), illetve tejsav hozzaadasat kovetően. A Lactobacillus plantarum 2142 SMSz hatekonyan csokkentette az IL-8 es a TNF-? szintjet illetve novelte a Hsp70 genexpressziojat. Kimutattuk, hogy a gyulladascsokkentő hatas alapja nem a feluluszo es a hidrogen-peroxid kozotti kolcsonhatas, ezert a Lactobacillus plantarum 2142 gyulladascsokkentő hatasanak alapja nem az SMSz peroxid bontasa. ...
Journal of the International Society of Antioxidants in Nutrition & Health, 2016
Cytochrome P450 enzymes have a remarkable role in the xenobiotics metabolism. Oxidation of drugs ... more Cytochrome P450 enzymes have a remarkable role in the xenobiotics metabolism. Oxidation of drugs and food components catalysed by the same isoenzyme can lead to drug-food interactions. Greatest quantity of CYP enzymes can be found in the liver. Nevertheless, the intestinal CYP garniture also contributes to the metabolism of clinically important drugs. Our research focused on the effects of flavonoids on the intestinal CYP enzymes. Intestinal epithelial cells (IPEC-J2) were treated by apigenin and apigenin-trimethylether. Phenobarbital was used as CYP inducer, mixture of naphtoflavone and ketoconazole as inhibitor. The CYP1A1, CYP1A2 and CYP3A activities were measured by chemiluminescent assay. Treatment of enterocytes with antipyrine was also performed in order to test possible drug interactions. Both flavones worked as significant CYP3A4 inhibitors. There was no significant difference between the inhibitory effects of the flavones at the same dose. However, apigenin-trimethylether ...
Background Chlorine dioxide (ClO2) is an inorganic, potent biocide and is available in highly pur... more Background Chlorine dioxide (ClO2) is an inorganic, potent biocide and is available in highly purified aqueous solution. It can be administered as an oral antiseptic in this form. Objectives Our aim is to determine the level of inflammatory markers and cytochrome genes expressed by enterocytes exposed to different concentrations of hyperpure chlorine dioxide solution. Methods Porcine jejunal enterocyte cell (IPEC‐J2) cultures were treated with the aqueous solution of hyper‐pure chlorine dioxide of various concentrations. We determined the alterations in mRNA levels of inflammatory mediators, such as IL6, CXCL8/IL8, TNF, HSPA6 (Hsp70), CAT and PTGS2 (COX2); furthermore, the expression of three cytochrome genes (CYP1A1, CYP1A2, CYP3A29) were analysed by quantitative PCR method. Results The highest applied ClO2 concentration reduced the expression of all three investigated CYP genes. The gene expression of PTGS2 and CAT were not altered by most concentrations of ClO2. The expression of IL8 gene was reduced by all applied concentrations of ClO2. TNF mRNA level was also decreased by most ClO2 concentrations used. Conclusions Different concentrations of chlorine dioxide exhibited immunomodulatory activity and caused altered transcription of CYP450 genes in porcine enterocytes. Further studies are needed to determine the appropriate ClO2 concentration for oral use in animals.
Orally applied butyrate as a nutritional supplement has a huge impact on the health of the gastro... more Orally applied butyrate as a nutritional supplement has a huge impact on the health of the gastrointestinal epithelium and the gut microflora. In addition, as an epigenetically active molecule, it can also influence the expression of certain genes. Regarding the results of some in vitro studies, butyrate, absorbed from the intestines and taken to the liver by the portal circulation, may alter the action of hepatic microsomal cytochrome P450 (CYP) enzymes, involved in the biotransformation of xenobiotics, having a remarkable importance in food safety, as well. The authors aimed to study the possible effects of orally applied butyrate on hepatic CYP activity in rats after weaning, also serving as a monogastric mammalian model. Animals were fed with normal pelleted stock diet without or with sodium butyrate (1.5 g/kg diet) for 21 days. As a positive control, a group of rats was treated with intraperitoneal phenobarbital injection. Animals were euthanized on day 21, microsomal fractions were separated from the exsanguinated livers by differential centrifugation. The activity of the most important (VP subfamilies was screened by specific enzyme assays (aminopyrine N-demethylation: CYP2B/3A, testosterone 6 beta-hydroxylation: CYP3A). According to the results, butyrate as a feed supplement did not cause any changes in the drug-metabolising activity of the examined hepatic microsomal CYP enzymes. However, phenobarbital significantly (P<0,05) increased the microsomal CYP2B and CYP3A activity of the liver. These data indicate that the supplementation of the diet with butyrate probably may not have any pharmacokinetic interactions with simultaneously applied xenobiotics in monogastric mammals, so it can be applied safely as a nutritional supplement.
To investigate whether the use of a novel synthetic medium in conjunction with impedimetric techn... more To investigate whether the use of a novel synthetic medium in conjunction with impedimetric technology could provide a rapid and automated detection of Pseudomonas aeruginosa in water samples. A selective synthetic medium (Z-broth) in which the only carbon and nitrogen source is acetamide was applied in direct impedimetric examination for the selective isolation of P. aeruginosa. A total of 1036 tap-water, well-water, swimming-pool water and dialysis water samples were investigated, and any P. aeruginosa contamination was detected in 7-24 h. Neither false-negative nor false-positive results were observed. The results of the present evaluation demonstrate that impedance measurement with the use of Z-broth is suitable for the rapid and automatic detection of P. aeruginosa in water. The main advantages of the method: 240 samples can be examined in one step, the procedure is fully automated, the results are obtained quickly and the labour and media requirements are low.
Journal of Veterinary Pharmacology and Therapeutics, 1994
Tiamulin selectively inhibits oxidative hepatic steroid and drug metabolism in vitro in the pig.]... more Tiamulin selectively inhibits oxidative hepatic steroid and drug metabolism in vitro in the pig.]. vet. Pharmacol. Therap. 17. 317-322. The simultaneous use of the antibiotic tiamulin with certain ionophoric antibiotics (monensin, salinomycin) may give rise to a toxic interaction in pigs and poultry. In the present study, effects of tiamulin on hepatic cytochrome P450 activities in vitro were studied using pig liver microsomes. When tiamulin was added to the incubation medium the N-demethylation rate of ethylmorphine and the hydroxylation of testosterone at the 68-and 1 la-positions was sirongly inhibited. Tiamulin inhibited these activities more than SKF525A or cimetidine, but less than ketoconazole. The microsomal N-demethylation rate of erythromycin and the hydroxylation of testosterone at the 28-position were inhibited to a lesser degree, whereas the ethoxyresorufin-0-deethylation, aniline hydroxylation and testosterone hydroxylations at the 15a-and 158-positions were not affected by tiamulin. No in vitro complexation by tiamulin of cytochrome P450 resulting in a loss of CO-binding capacity could be demonstrated. Results from the present study suggest a selective inhibition of cytochrome P450 enzymes in pigs, probably belonging to the P4503A subfamily. The mechanism of this interaction is still unclear. However, interactions between tiamulin and those veterinary drugs or endogenous compounds which undergo oxidative metabolism by P450 enzymes must be considered. More research is needed to reveal which of the P450 enzymes are affected by tiamulin in order to improve the understanding and probably the predictability of this interaction.
Food samples were monitored for contamination with Listeria monocytogenes, and the incidence of h... more Food samples were monitored for contamination with Listeria monocytogenes, and the incidence of human listeriosis was evaluated according to the data obtained in Hungary in the year 2004. Of the food samples tested, the bacterium was most often detectable in milk and dairy products, as 72.1% of all L. monocytogenes strains were isolated from these samples. The food samples most commonly yielded strains of serotype 1/2a (45.1%) and 4b (27.0%). In 2004, 3 perinatal and 14 nonperinatal human listeriosis cases were diagnosed in Hungary. These disease cases were most often caused by strains belonging to serotype 4b (52.8%) and serotype 1/2a (23.5%). On the basis of the antibiotic sensitivity test results of strains isolated from the disease cases, penicillin and aminoglycoside antibiotics or a combination thereof were found to be effective.
A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 e... more A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 epithelial cells as upper layer, was developed to study the effects of lipopolysaccharides (LPS) on the gene expression profile of pro-inflammatory cytokines (interleukin-8 (IL-8) and tumor necrosis factor-α) and CYP enzymes (CYP1A1, CYP1A2, CYP3A29). The barrier integrity of IPEC-J2 cells was investigated by transepithelial electrical resistance measurements and by fluorescein isothiocyanate-dextran-based test. Basolateral IL-8 production was significantly elevated in LPS-treated IPEC-J2 and primary hepatocyte mono-cultures as well as in the co-culture system, in a dose-independent manner. The LPS-induced changes in the expression of the CYP1A2 and CYP3A29 genes in hepatocyte mono-cultures differed from those in co-culture after LPS treatment on the apical side of the IPEC-J2 cell layer. CYP1A2 was downregulated by the LPS treatment in mono-cultures but upregulated at 10 μg/ml LPS in co-culture; gene expression of CYP3A29 showed no significant LPS-induced change in the hepatocyte mono-culture but was significantly downregulated in co-culture. The newly established co-culture system capable of mimicking enterohepatic interplay in LPS-induced inflammatory responses in vitro can be used in the future for reliable screening of potential anti-inflammatory compounds.
To investigate the role of reactive oxygen species (ROS) induced by butyrate in tumor cells, we c... more To investigate the role of reactive oxygen species (ROS) induced by butyrate in tumor cells, we compared HT29R, an HT29-derived human colon cancer cell line refractory to butyrate-induced cell differentiation but highly sensitive to cell death, with the differentiationpositive HT29-12 and HT29-21 cell lines (exhibiting low sensitivity to butyrate-induced cell death), with respect to levels of butyrate-induced free radicals (FRs), ROS, and H 2 O 2. Dose-dependent increase of FRs (as determined by electron spin resonance spectroscopy) and ROS (dichlorofluorescein assay) was induced in HT29R, but not in HT29-12 and HT29-21 cells, where, in contrast to HT29R, a dose-dependent increase of H 2 O 2 release (phenol red assay) was induced by butyrate. The mode of butyrateinduced cell death in HT29R cells was of a mixed type with necrosis predominating, which, however, switched to apoptosis as the major type of cell death in the presence of the drugs 1,5-dihydroxyisoquinoline, resveratrol, or cyclosporine A. The results suggest that FRs and ROS induced by butyrate in HT29R cells are products of cell death, while H 2 O 2 induced in HT29-12 and HT29-21 cells is functionally related to cell differentiation. Keywords Butyrate Á Colon cancer Á HT29 Á Differentiation Á Cell death Á ROS
This study was based on our previously developed double-layered enterohepatic co-culture system, ... more This study was based on our previously developed double-layered enterohepatic co-culture system, composed of nontumorigenic porcine intestinal epithelial cell line (IPEC-J2) and primary culture of porcine hepatocytes. The anti-inflammatory effect of spent culture supernatant of Lactobacillus plantarum 2142 (Lp2142; 13.3%) and sodium n-butyrate (2 mM) was tested on IPEC-J2 and hepatocyte monocultures as well as on the gut-liver co-culture. To mimic inflammation, lipopolysaccharide (LPS; 1 and 10 μg/mL) was applied. Production of IL-8 and IL-6 was measured as a marker of inflammatory responses. The paracellular permeability of the intestinal epithelium was also monitored by fluoresceinisothiocyanate-labeled dextran 4 assay. Significant increase of IL-8 concentration was observed in the IPEC-J2 monoculture (P < 0.01) while the level of IL-6 was not changed following LPS treatment. Concentration of IL-8 and IL-6 was grown significantly in hepatocyte monocultures (P < 0.05 and P < 0.001) as well as in the co-culture after 10 μg/mL LPS treatment (P < 0.001 and P < 0.001). One microgram per milliliter LPS caused elevated IL-8 level in the co-culture (P < 0.001) and in the hepatocyte monoculture (P < 0.01), while it caused increased IL-6 level only in the hepatocytes (P < 0.001). Production of IL-8 was significantly decreased by butyrate in case of 1 μg/mL as well as 10 μg/mL LPS exposure in the co-culture (P < 0.001). Application of butyrate also reduced IL-6 level in the co-culture after 10 μg/ mL LPS treatment (P < 0.01). Lactobacillus plantarum 2142 decreased IL-8 level after incubation with 1 μg/ mL LPS (P < 0.001), while in case of 10 μg/mL LPS treatment only a marginal lowering in IL-8 (P = 0.064) release was measured. The IL-6 concentration was significantly reduced (P < 0.01 in case of 1 μg/mL LPS treatment) by Lp2142 in the co-culture. Contrarily, the elevated IL-8 and IL-6 level of hepatocytes has not been reduced in case of either butyrate or Lp2142 addition. The enterohepatic co-culture model offers a possibility for fast and reliable screening of new candidates against enteric inflammation, which are of special interest in porcine medicine and health management. According to our results, Lp2142 and butyrate both seem to be effective as anti-inflammatory agents in LPS-triggered inflammatory response, tested in the gut-liver co-culture model.
This article is an open access article distributed under the terms and conditions of the Creative... more This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY
The authors investigated the possibility of the presence of VTEC strains in improperly pasteurize... more The authors investigated the possibility of the presence of VTEC strains in improperly pasteurized milk samples. A total of 64 Escherichia coli strains were isolated from 135 pasteurized milk samples originating from the same producer. The examined isolates contained 29 haemolysin-, 9 colicin-and 5 aerobactin-producing strains, but the investigations concerning heat-resistant and heat-sensitive toxins gave negative results. Six O128-type E. coli strains exerted a cytotoxic effect on the VERO cell line; 5 of them contained H12 antigen, while one could not be typed. Four of the 6 verocytotoxin-producing strains belonged in phage group 20, one in phage group (2)3(7), and one in phage group 4; four strains were of B3, one of A1, and one of A1(A2) phage type. Because of a technical failure the milk was pasteurized at 69 ºC for 15 s, which is 2 ºC less than required. The results underline the importance of the appropriate pasteurization temperature, as otherwise the milk may contain verocytotoxin-producing E. coli, which is a potentially great hazard for public health.
Cytochrome P450 (CYP) oxidases are among the main metabolizing enzymes that are responsible for t... more Cytochrome P450 (CYP) oxidases are among the main metabolizing enzymes that are responsible for the transformation of xenobiotics, including clinically important drugs. Their activity can be influenced by several compounds leading to decreased efficacy or increased toxicity of co-administered medicines. Flavonoids exert various beneficial effects on human and animal health; therefore they are used as food and feed supplements. However, they are also well-known for their CYP modulating potential. Since the amount of CYP enzymes is highest in the liver, interaction studies are mainly conducted in hepatocytes, however, CYP activity in the gastrointestinal tract is also remarkable. In this study, effects of apigenin (API), quercetin (QUE) and their methylated derivatives trimethylapigenin (TM-API), 3-O-methylquercetin (3M-QUE) and 3′,7-di-O-methylquercetin (3′7DM-QUE) on the CYP enzyme activity was examined in IPEC-J2 porcine intestinal epithelial cells. Potential food-drug interactions...
Journal of Veterinary Pharmacology and Therapeutics, 2014
Butyrate, a commonly applied feed additive in poultry nutrition, can modify the expression of cer... more Butyrate, a commonly applied feed additive in poultry nutrition, can modify the expression of certain genes, including those encoding cytochrome P450 (CYP) enzymes. In comparative in vitro and in vivo experiments, the effect of butyrate on hepatic CYP genes was examined in primary cultures of chicken hepatocytes and in liver samples of chickens collected from animals that had been given butyrate as a feed additive. Moreover, the effect of butyrate on the biotransformation of erythromycin, a marker substance for the activity of enzymes of the CYP3A family, was investigated in vitro and in vivo. Butyrate increased the expression of the avian-specific CYP2H1 both in vitro and in vivo. In contrast, the avian CYP3A37 expression was decreased in hepatocytes following butyrate exposure, but not in the in vivo model. CYP1A was suppressed by butyrate in the in vitro experiments, and overexpressed in vivo in butyrate-fed animals. The concomitant incubation of hepatocytes with butyrate and erythromycin led to an increased CYP2H1 expression and a less pronounced inhibition of CYP3A37. In in vivo pharmacokinetic experiments, butyrate-fed animals given a single i.m. injection of erythromycin, a slower absorption phase (longer T(half-abs) and delayed T(max)) but a rapid elimination phase of this marker substrate was observed. Although these measurable differences were detected in the pharmacokinetics of erythromycin, it is unlikely that a concomitant application of sodium butyrate with erythromycin or other CYP substrates will cause clinically significant feed-drug interaction in chickens.
Effects of orally applied butyrate bolus on histone me P450 enzyme activity Conclusion: Orally ad... more Effects of orally applied butyrate bolus on histone me P450 enzyme activity Conclusion: Orally added butyrate in bolus could cause in vivo hyperacetylation of the hepatic core histones, Mátis et al. Nutrition & Metabolism 2013, 10:12
Az akut oxidativ stressz es a probiotikumok hatasanak vizsgalatahoz kollagennel bevont polieszter... more Az akut oxidativ stressz es a probiotikumok hatasanak vizsgalatahoz kollagennel bevont polieszter membran inzerten tenyesztett, ujszulott sertes jejunum eredetű IPEC-J2 sejtvonalat alkalmaztunk. A peroxiddal kezelt IPEC-J2 sejtekben a gyulladasos citokinek (IL-8 es TNF-?) es a vedő hatasu hősokk feherje 70 (Hsp 70) relativ genexpressziojat kvantitativ real-time PCR-rel hataroztuk meg a probiotikus bakteriumtorzsek (Lactobacillus plantarum 2142, Lactobacillus casei Shirota, Bifidobacterium animalis subsp. lactis BB-12, Bacillus amyloliquefaciens CECT 5940, Enterococcus faecium CECT 4515) feluluszojanak (SMSz), illetve tejsav hozzaadasat kovetően. A Lactobacillus plantarum 2142 SMSz hatekonyan csokkentette az IL-8 es a TNF-? szintjet illetve novelte a Hsp70 genexpressziojat. Kimutattuk, hogy a gyulladascsokkentő hatas alapja nem a feluluszo es a hidrogen-peroxid kozotti kolcsonhatas, ezert a Lactobacillus plantarum 2142 gyulladascsokkentő hatasanak alapja nem az SMSz peroxid bontasa. ...
Journal of the International Society of Antioxidants in Nutrition & Health, 2016
Cytochrome P450 enzymes have a remarkable role in the xenobiotics metabolism. Oxidation of drugs ... more Cytochrome P450 enzymes have a remarkable role in the xenobiotics metabolism. Oxidation of drugs and food components catalysed by the same isoenzyme can lead to drug-food interactions. Greatest quantity of CYP enzymes can be found in the liver. Nevertheless, the intestinal CYP garniture also contributes to the metabolism of clinically important drugs. Our research focused on the effects of flavonoids on the intestinal CYP enzymes. Intestinal epithelial cells (IPEC-J2) were treated by apigenin and apigenin-trimethylether. Phenobarbital was used as CYP inducer, mixture of naphtoflavone and ketoconazole as inhibitor. The CYP1A1, CYP1A2 and CYP3A activities were measured by chemiluminescent assay. Treatment of enterocytes with antipyrine was also performed in order to test possible drug interactions. Both flavones worked as significant CYP3A4 inhibitors. There was no significant difference between the inhibitory effects of the flavones at the same dose. However, apigenin-trimethylether ...
Background Chlorine dioxide (ClO2) is an inorganic, potent biocide and is available in highly pur... more Background Chlorine dioxide (ClO2) is an inorganic, potent biocide and is available in highly purified aqueous solution. It can be administered as an oral antiseptic in this form. Objectives Our aim is to determine the level of inflammatory markers and cytochrome genes expressed by enterocytes exposed to different concentrations of hyperpure chlorine dioxide solution. Methods Porcine jejunal enterocyte cell (IPEC‐J2) cultures were treated with the aqueous solution of hyper‐pure chlorine dioxide of various concentrations. We determined the alterations in mRNA levels of inflammatory mediators, such as IL6, CXCL8/IL8, TNF, HSPA6 (Hsp70), CAT and PTGS2 (COX2); furthermore, the expression of three cytochrome genes (CYP1A1, CYP1A2, CYP3A29) were analysed by quantitative PCR method. Results The highest applied ClO2 concentration reduced the expression of all three investigated CYP genes. The gene expression of PTGS2 and CAT were not altered by most concentrations of ClO2. The expression of IL8 gene was reduced by all applied concentrations of ClO2. TNF mRNA level was also decreased by most ClO2 concentrations used. Conclusions Different concentrations of chlorine dioxide exhibited immunomodulatory activity and caused altered transcription of CYP450 genes in porcine enterocytes. Further studies are needed to determine the appropriate ClO2 concentration for oral use in animals.
Orally applied butyrate as a nutritional supplement has a huge impact on the health of the gastro... more Orally applied butyrate as a nutritional supplement has a huge impact on the health of the gastrointestinal epithelium and the gut microflora. In addition, as an epigenetically active molecule, it can also influence the expression of certain genes. Regarding the results of some in vitro studies, butyrate, absorbed from the intestines and taken to the liver by the portal circulation, may alter the action of hepatic microsomal cytochrome P450 (CYP) enzymes, involved in the biotransformation of xenobiotics, having a remarkable importance in food safety, as well. The authors aimed to study the possible effects of orally applied butyrate on hepatic CYP activity in rats after weaning, also serving as a monogastric mammalian model. Animals were fed with normal pelleted stock diet without or with sodium butyrate (1.5 g/kg diet) for 21 days. As a positive control, a group of rats was treated with intraperitoneal phenobarbital injection. Animals were euthanized on day 21, microsomal fractions were separated from the exsanguinated livers by differential centrifugation. The activity of the most important (VP subfamilies was screened by specific enzyme assays (aminopyrine N-demethylation: CYP2B/3A, testosterone 6 beta-hydroxylation: CYP3A). According to the results, butyrate as a feed supplement did not cause any changes in the drug-metabolising activity of the examined hepatic microsomal CYP enzymes. However, phenobarbital significantly (P<0,05) increased the microsomal CYP2B and CYP3A activity of the liver. These data indicate that the supplementation of the diet with butyrate probably may not have any pharmacokinetic interactions with simultaneously applied xenobiotics in monogastric mammals, so it can be applied safely as a nutritional supplement.
To investigate whether the use of a novel synthetic medium in conjunction with impedimetric techn... more To investigate whether the use of a novel synthetic medium in conjunction with impedimetric technology could provide a rapid and automated detection of Pseudomonas aeruginosa in water samples. A selective synthetic medium (Z-broth) in which the only carbon and nitrogen source is acetamide was applied in direct impedimetric examination for the selective isolation of P. aeruginosa. A total of 1036 tap-water, well-water, swimming-pool water and dialysis water samples were investigated, and any P. aeruginosa contamination was detected in 7-24 h. Neither false-negative nor false-positive results were observed. The results of the present evaluation demonstrate that impedance measurement with the use of Z-broth is suitable for the rapid and automatic detection of P. aeruginosa in water. The main advantages of the method: 240 samples can be examined in one step, the procedure is fully automated, the results are obtained quickly and the labour and media requirements are low.
Journal of Veterinary Pharmacology and Therapeutics, 1994
Tiamulin selectively inhibits oxidative hepatic steroid and drug metabolism in vitro in the pig.]... more Tiamulin selectively inhibits oxidative hepatic steroid and drug metabolism in vitro in the pig.]. vet. Pharmacol. Therap. 17. 317-322. The simultaneous use of the antibiotic tiamulin with certain ionophoric antibiotics (monensin, salinomycin) may give rise to a toxic interaction in pigs and poultry. In the present study, effects of tiamulin on hepatic cytochrome P450 activities in vitro were studied using pig liver microsomes. When tiamulin was added to the incubation medium the N-demethylation rate of ethylmorphine and the hydroxylation of testosterone at the 68-and 1 la-positions was sirongly inhibited. Tiamulin inhibited these activities more than SKF525A or cimetidine, but less than ketoconazole. The microsomal N-demethylation rate of erythromycin and the hydroxylation of testosterone at the 28-position were inhibited to a lesser degree, whereas the ethoxyresorufin-0-deethylation, aniline hydroxylation and testosterone hydroxylations at the 15a-and 158-positions were not affected by tiamulin. No in vitro complexation by tiamulin of cytochrome P450 resulting in a loss of CO-binding capacity could be demonstrated. Results from the present study suggest a selective inhibition of cytochrome P450 enzymes in pigs, probably belonging to the P4503A subfamily. The mechanism of this interaction is still unclear. However, interactions between tiamulin and those veterinary drugs or endogenous compounds which undergo oxidative metabolism by P450 enzymes must be considered. More research is needed to reveal which of the P450 enzymes are affected by tiamulin in order to improve the understanding and probably the predictability of this interaction.
Food samples were monitored for contamination with Listeria monocytogenes, and the incidence of h... more Food samples were monitored for contamination with Listeria monocytogenes, and the incidence of human listeriosis was evaluated according to the data obtained in Hungary in the year 2004. Of the food samples tested, the bacterium was most often detectable in milk and dairy products, as 72.1% of all L. monocytogenes strains were isolated from these samples. The food samples most commonly yielded strains of serotype 1/2a (45.1%) and 4b (27.0%). In 2004, 3 perinatal and 14 nonperinatal human listeriosis cases were diagnosed in Hungary. These disease cases were most often caused by strains belonging to serotype 4b (52.8%) and serotype 1/2a (23.5%). On the basis of the antibiotic sensitivity test results of strains isolated from the disease cases, penicillin and aminoglycoside antibiotics or a combination thereof were found to be effective.
A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 e... more A porcine enterohepatic co-culture system, with primary hepatocytes as bottom layer and IPEC-J2 epithelial cells as upper layer, was developed to study the effects of lipopolysaccharides (LPS) on the gene expression profile of pro-inflammatory cytokines (interleukin-8 (IL-8) and tumor necrosis factor-α) and CYP enzymes (CYP1A1, CYP1A2, CYP3A29). The barrier integrity of IPEC-J2 cells was investigated by transepithelial electrical resistance measurements and by fluorescein isothiocyanate-dextran-based test. Basolateral IL-8 production was significantly elevated in LPS-treated IPEC-J2 and primary hepatocyte mono-cultures as well as in the co-culture system, in a dose-independent manner. The LPS-induced changes in the expression of the CYP1A2 and CYP3A29 genes in hepatocyte mono-cultures differed from those in co-culture after LPS treatment on the apical side of the IPEC-J2 cell layer. CYP1A2 was downregulated by the LPS treatment in mono-cultures but upregulated at 10 μg/ml LPS in co-culture; gene expression of CYP3A29 showed no significant LPS-induced change in the hepatocyte mono-culture but was significantly downregulated in co-culture. The newly established co-culture system capable of mimicking enterohepatic interplay in LPS-induced inflammatory responses in vitro can be used in the future for reliable screening of potential anti-inflammatory compounds.
To investigate the role of reactive oxygen species (ROS) induced by butyrate in tumor cells, we c... more To investigate the role of reactive oxygen species (ROS) induced by butyrate in tumor cells, we compared HT29R, an HT29-derived human colon cancer cell line refractory to butyrate-induced cell differentiation but highly sensitive to cell death, with the differentiationpositive HT29-12 and HT29-21 cell lines (exhibiting low sensitivity to butyrate-induced cell death), with respect to levels of butyrate-induced free radicals (FRs), ROS, and H 2 O 2. Dose-dependent increase of FRs (as determined by electron spin resonance spectroscopy) and ROS (dichlorofluorescein assay) was induced in HT29R, but not in HT29-12 and HT29-21 cells, where, in contrast to HT29R, a dose-dependent increase of H 2 O 2 release (phenol red assay) was induced by butyrate. The mode of butyrateinduced cell death in HT29R cells was of a mixed type with necrosis predominating, which, however, switched to apoptosis as the major type of cell death in the presence of the drugs 1,5-dihydroxyisoquinoline, resveratrol, or cyclosporine A. The results suggest that FRs and ROS induced by butyrate in HT29R cells are products of cell death, while H 2 O 2 induced in HT29-12 and HT29-21 cells is functionally related to cell differentiation. Keywords Butyrate Á Colon cancer Á HT29 Á Differentiation Á Cell death Á ROS
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Papers by György Csikó