Fusarium-derived mycotoxin deoxynivalenol (DON) usually induces diarrhea, vomiting and gastrointestinal inflammation. We studied the cytotoxic effect of DON on porcine small intestinal epithelium using the intestinal porcine epithelial cell line IPEC-J2. We screened out differentially expressed genes (DEGs) using RNA-seq and identified 320 upregulated genes and 160 downregulated genes. The enrichment pathways of these DEGs focused on immune-related pathways. DON induced proinflammatory gene expression, including cytokines, chemokines and other inflammation-related genes. DON increased IL1A, IL6 and TNF-α release and DON activated the phosphorylation of extracellular signal-regulated kinase-1 and-2 (ERK1/2), JUN N-terminal kinase (JNK) and p38 MAPK. A p38 inhibitor attenuated DON-induced IL6, TNF-α, CXCL2, CXCL8, IL12A, IL1A, CCL20, CCL4 and IL15 production, while an ERK1/2 inhibitor had only a small inhibitory effect on IL15 and IL6. An inhibitor of p38 MAPK decreased the release of IL1A, IL6 and TNF-α and an inhibitor of ERK1/2 partly attenuated protein levels of IL6. These data demonstrate that DON induces proinflammatory factor production in IPEC-J2 cells by activating p38 and ERK1/2.

• MeSH
• buněčné linie MeSH
• epitelové buňky účinky léků   imunologie   metabolismus   MeSH
• interleukin-1 genetika   MeSH
• interleukin-6 genetika   MeSH
• MAP kinasový signální systém účinky léků   genetika   imunologie   MeSH
• mitogenem aktivované proteinkinasy p38 metabolismus   MeSH
• prasata MeSH
• střevní sliznice účinky léků   imunologie   metabolismus   MeSH
• TNF-alfa genetika   MeSH
• transkriptom účinky léků   MeSH
• trichotheceny toxicita   MeSH
• viabilita buněk účinky léků   MeSH
• zánět MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

This study was to evaluate the efficacy of TOXO-XL (XL), an integrated mycotoxin-mitigating agent, on aflatoxin B1 (AFB1)-induced damage in Leghorn male hepatoma (LMH), porcine jejunum epithelial cell line (IPEC-J2) and porcine alveolar macrophages (3D4/21) cells, and to explore its potential mechanisms. The results showed that 30% inhibition concentration (IC30) of AFB1 in LMH, IPEC-J2 and 3D4/21 cells was 0.5, 15.0, and 2.5 mg/L, respectively. Notably, cell viability, ROS, apoptosis and DNA lesion induced by AFB1 (IC30) could be ameliorated by the supplementation with XL at the dosage of 0.025, 0.025 and 0.005%, respectively. Additionally, the migration and phagocytosis abilities impaired by AFB1 were also restored by XL in 3D4/21. Further experiments revealed that XL supplementation markedly attenuated AFB1-induced inflammatory response by decreasing IL-1β, IL-6 and IL-10 in LMH, IL-6 in IPEC-J2 and IL-1β in 3D4/21 cells. Meanwhile, XL supplementation reversed the alterations of BAX, BCL-2 and caspase-3 induced by AFB1 in the three cells, suggesting that AFB1-induced apoptosis may be suppressed via the mitochondria-dependent pathway. Furthermore, XL may have a protective effect on the intestinal barrier through the restoration of occludin protein. Conclusively, these findings indicated that XL could alleviate AFB1-induced cytotoxicity in the three cells, potentially through the regulation of cytokines, ROS, apoptotic and DNA damage signaling.

• MeSH
• aflatoxin B1 toxicita   metabolismus   MeSH
• apoptóza MeSH
• epitelové buňky MeSH
• hepatocelulární karcinom * metabolismus   MeSH
• interleukin-6 metabolismus   MeSH
• kur domácí metabolismus   MeSH
• nádory jater * metabolismus   MeSH
• prasata MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

This study aimed to evaluate the immunomodulatory effect of the probiotic Limosilactobacillus reuteri L26 BiocenolTM (L26) and its purified exopolysaccharide (EPS) with respect to antiviral innate immune response. In our experiment, we used porcine epithelial IPEC-J2 cells as a model of the intestinal barrier in a homologous infection by porcine Rotavirus A strain OSU6 (RVA). The production of selected molecules of non-specific humoral immunity was evaluated at the mRNA level. The EPS alone significantly increased the level of interferon λ3 (IFN-λ3) mRNA in the non-infected IPEC-J2 cells (P < 0.001). We also tested whether the treatment of IPEC-J2 cells by L26 or EPS influences the replication of RVA by virus titration and real-time PCR. We found that a pre-treatment in combination with subsequent continuous stimulation has no influence on the RVA replication. However, both treatments significantly decreased the RVA-induced production of IFN-λ3 (P < 0.05) and the “SOS” cytokine interleukin 6 (IL-6; P < 0.01), already at the transcription level. In addition, the EPS treatment resulted in significantly increased IL-10 mRNA in the RVA-infected cells. In summary, we assume an immunoregulatory potential of L. reuteri L26 BiocenolTM and its EPS in the local intestinal antiviral immune response.

The aim of the study was to broadly determine the biological activities of purple potato ethanolic extract of the Blue Congo variety (BCE). The antioxidant activity of BCE was determined in relation to liposome membranes, and peroxidation was induced by UVB and AAPH. To clarify the antioxidant activity of BCE, we investigated its interactions with hydrophilic and hydrophobic regions of a membrane using fluorimetric and FTIR methods. Next, we investigated the cytotoxicity and pro-apoptotic activities of BCE in two human colon cancer cell lines (HT-29 and Caco-2) and in normal cells (IPEC-J2). In addition, the ability to inhibit enzymes that are involved in pro-inflammatory reactions was examined. Furthermore, BCE interactions with serum albumin and plasmid DNA were investigated using steady state fluorescence spectroscopy and a single molecule fluorescence technique (TCSPC-FCS). We proved that BCE effectively protects lipid membranes against the process of peroxidation and successfully inhibits the cyclooxygenase and lipoxygenase enzymes. Furthermore, it interacts with the hydrophilic and hydrophobic parts of lipid membranes as well as with albumin and plasmid DNA. It was observed that BCE is more cytotoxic against colon cancer cell lines than normal IPEC-J2 cells; it also induces apoptosis in cancer cell lines, but does not induce cell death in normal cells.

• MeSH
• albuminy MeSH
• antioxidancia chemie   farmakologie   MeSH
• fytogenní protinádorové látky chemie   farmakologie   MeSH
• inhibitory cyklooxygenasy chemie   farmakologie   MeSH
• inhibitory lipoxygenas chemie   farmakologie   MeSH
• lidé MeSH
• lipidy chemie   MeSH
• liposomy MeSH
• nádorové buněčné linie MeSH
• plazmidy MeSH
• reaktivní formy kyslíku MeSH
• rostlinné extrakty chemie   farmakologie   MeSH
• sérový albumin chemie   metabolismus   MeSH
• Solanum tuberosum chemie   MeSH
• vazba proteinů MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

In this study we have compared protein secretion in the wild type of S. Typhimurium and the rfaC mutant. We found out that the rfaC mutant was defective in protein secretion. In addition, the rfaC mutant was defective in its invasion into an IPEC-J2 porcine epithelial cell line and also in motility in semisolid agar. Consistent with this, reduced flagella numbers were observed in the rfaC mutant. In the rfaC mutant, there were no defects in flagellin expression as detected by western blot and immune electron microscopy which demonstrated equal amounts of flagellin in the cytoplasm of both the rfaC mutant and the wild-type S. Typhimurium. However, in the wild-type strain only, the flagellin was assembled to spatially restricted areas on the inner side of cytoplasmic membrane. The oligosaccharide core of LPS is therefore required for the assembly of flagella and T3SS secretion machinery followed by protein secretion.

• MeSH
• bakteriální sekreční systémy MeSH
• buněčné linie MeSH
• cytoplazma chemie   MeSH
• epitelové buňky mikrobiologie   MeSH
• flagelin biosyntéza   sekrece   MeSH
• flagella metabolismus   MeSH
• imunoelektronová mikroskopie MeSH
• lipopolysacharidy chemie   MeSH
• mutace MeSH
• prasata MeSH
• Salmonella enterica genetika   metabolismus   ultrastruktura   MeSH
• sekvenční analýza hybridizací s uspořádaným souborem oligonukleotidů MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

ANNOTATION 1 Analytical gas chromatography methods (GC/FID), (GC/ECD) and (GC/MS) for alcoxycarboxylic acids (ethoxyacetic and butoxyacetic acid) determination using modern derivatization reagents will be drawn up. ANNOTATION 2 System of internal qualitycontrol will be assured by preparation of pooled freeze-dryied urine sample of persons occupationally exposed to glycolethers.The values of alcoxycarboxylic acids concentrations and their uncertainties will be evaluated using statistical program IPECA.ANNOTATION 3 The results will serve for the collection of the background data for the categorization of occupations and for updating the Threshold Limits Values of Biological Indicators in Czech Republic.

ANOTACE 1 Budou vypracovány analytické metody pro stanovení biomarkerů expozice glykoletherům plynovou chromatografií s detekcí plamenoionizační, hmotnostní a detekcí elektronového záchytu s využitím moderních derivatizačních činidel. ANOTACE 2 Systém vnitřní kontroly kvality bude zajištěn přípravou lyofilizovaného vzorku moče exponovaných osob. Hodnoty koncentrace alkoxykarboxylových kyselin a jejich nejistoty budou vypočteny statistickým programem IPECA v souladu s EU normami. ANOTACE 3 Metody budou využity v rámci preventivních prohlídek a k získávání podkladů pro kategorizaci prací. Výsledky studie budou sloužit jako podklady pro úpravu limitních hodnot škodlivých faktorů expozice glykoletherům v pracovním prostředí.

• MeSH
• acetáty analýza   moč   toxicita   MeSH
• biologické markery MeSH
• chromatografie plynová metody   využití   MeSH
• ethylenglykoly analýza   moč   toxicita   MeSH
• inhalační expozice MeSH
• kožní absorpce MeSH
• monitorování životního prostředí MeSH
• pracovní expozice analýza   normy   MeSH

• Konspekt
• Chemie. Mineralogické vědy
• NLK Obory
• chemie, klinická chemie
• pracovní lékařství
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR