Projekt bude zaměřen na detekci salmonel,kampylobakterů a L.monocytogenes z potravinového řetězce. Stanovení vybraných patogenů bude prováděno imunologickými metodami - ELISA a IMS. Pro diagnostiku salmonel bude vypracována a ověřena metoda založená na detekci specifického genu pomocí PCR.; Project will be intented on Samonella, Campylobacter and L.monocytogenes detection from food chain. Determination of selected pathogens will be performed by immunological methods - ELISA and IMS. For the diagnostic of Salmonella, a method will be prepared and verified based on the detection of a specific gene with the help of PCR.

• MeSH
• imunoanalýza využití   metody   MeSH
• imunochemie využití   metody   MeSH
• imunomagnetická separace využití   metody   MeSH
• kontaminace potravin analýza   MeSH
• latex fixační testy využití   metody   MeSH
• polymerázová řetězová reakce využití   metody   MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• nutriční terapie, dietoterapie a výživa
• chemie, klinická chemie
• biologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Mimotělní oběh /MTO/ u srdečních operací způsobuje systémovou zánětlivou resekci /SZR/, která poškozuje pacienta. Cílem studie bylo ovlivnit tuto reakci předoperační aferézou /PA/. Bylo zjištěno, že PA signifikantně snižuje SZR po MTO.

• MeSH
• endotoxiny MeSH
• hemoperfuze MeSH
• kardiochirurgické výkony MeSH
• mimotělní oběh MeSH
• pooperační komplikace prevence a kontrola   MeSH
• zánět prevence a kontrola   etiologie   MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• chirurgie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu IGA MZ ČR

Eleven strains of Lactobacillus collected in the Culture Collection of Dairy Microorganisms (CCDM) were evaluated for selected probiotic properties such as survival in gastrointestinal fluids, antimicrobial activity, and competition with non-toxigenic Escherichia coli O157:H7 for adhesion on Caco-2 cells. The viable count of lactobacilli was reduced during 3-h incubation in gastric fluid followed by 3-h incubation in intestinal fluid. All strains showed antimicrobial activity and the three most effective strains inhibited the growth of at least 16 indicator strains. Antimicrobial metabolites of seven strains active against Lactobacillus and Clostridium indicator strains were found to be sensitive to proteinase K and trypsin, which indicates their proteinaceous nature. The degree of competitive inhibition of non-toxigenic E. coli O157:H7 adhesion on the surface of Caco-2 cells was strain-dependent. A significant decrease (P < 0.05) in the number of non-toxigenic E. coli O157:H7 adhering to Caco-2 cells was observed with all lactobacilli. Three strains were selected for additional studies of antimicrobial activity, i.e., Lactobacillus gasseri CCDM 215, Lactobacillus acidophilus CCDM 149, and Lactobacillus helveticus CCDM 82.

• MeSH
• antibióza * MeSH
• antiinfekční látky metabolismus   MeSH
• bakteriální adheze MeSH
• Caco-2 buňky MeSH
• epitelové buňky mikrobiologie   MeSH
• Escherichia coli O157 fyziologie   MeSH
• Lactobacillus růst a vývoj   fyziologie   MeSH
• lidé MeSH
• mikrobiální viabilita účinky léků   MeSH
• probiotika farmakologie   MeSH
• žaludeční šťáva mikrobiologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakt z konference MeSH

Wedelolactone is one of the active plant polyphenolic compounds. Anti-tumor effects of this drug have been demonstrated recently. We have described that wedelolactone acts as catalytic inhibitor of DNA topoisomerase IIα. The aim of this study was to further characterize the mechanism of its anti-tumor effects. We showed that wedelolactone inhibits binding of DNA topoisomerase IIα to plasmid DNA and antagonizes formation of etoposide-induced DNA cleavage complex. The inhibition of topoisomerase IIα by wedelolactone is reversible by excess of the enzyme but not DNA. The in vitro inhibitory effect of wedelolactone on the topoisomerase IIα activity is redox-dependent as it diminished in the presence of reducing agents. Cytotoxicity of wedelolactone was partially inhibited by N-acetylcysteine and glutathione ethyl ester in breast cancer MDA-MB-231 and MDA-MB-468 cells while the inhibitory effect of catalase was observed only in the former cell line. Finally, we found that wedelolactone can be oxidized in the presence of copper ions resulting in DNA strand break and abasic site formation in vitro. However, wedelolactone induced neither DNA damage in MDA-MB-231 cells nor mutations in bacterial cells detectable by Ames test suggesting that wedelolactone may not be an effective inducer of DNA damage. We conclude that the topoisomerase IIα inhibitory- and DNA damaging activities of wedelolactone in vitro depend on its redox state. Pro-oxidant activity could, however, explain only part of wedelolactone-induced cytotoxicity. Therefore, the major cellular target(s) of wedelolactone and the exact mechanism of wedelolactone-induced cytotoxicity still remain to be identified.

• MeSH
• acetylcystein MeSH
• antigeny nádorové metabolismus   MeSH
• DNA vazebné proteiny antagonisté a inhibitory   metabolismus   MeSH
• DNA-topoisomerasy typu II metabolismus   MeSH
• glutathion analogy a deriváty   MeSH
• imunoblotting MeSH
• katalasa MeSH
• kumariny chemie   metabolismus   farmakologie   MeSH
• lidé MeSH
• molekulární struktura MeSH
• NAD metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádory prsu farmakoterapie   MeSH
• oxidace-redukce MeSH
• protinádorové látky chemie   metabolismus   farmakologie   MeSH
• techniky in vitro MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Bifidobacterium longum is considered to play an important role in health maintenance of the human gastrointestinal tract. Probiotic properties of bifidobacterial isolates are strictly strain-dependent and reliable methods for the identification and discrimination of this species at both subspecies and strain levels are thus required. Differentiation between B. longum ssp. longum and B. longum ssp. infantis is difficult due to high genomic similarities. In this study, four molecular-biological methods (species- and subspecies-specific PCRs, random amplified polymorphic DNA (RAPD) method using 5 primers, repetitive sequence-based (rep)-PCR with BOXA1R and (GTG)(5) primers and amplified ribosomal DNA restriction analysis (ARDRA)) and biochemical analysis, were compared for the classification of 30 B. longum strains (28 isolates and 2 collection strains) on subspecies level. Strains originally isolated from the faeces of breast-fed healthy infants (25) and healthy adults (3) showed a high degree of genetic homogeneity by PCR with subspecies-specific primers and rep-PCR. When analysed by RAPD, the strains formed many separate clusters without any potential for subspecies discrimination. These methods together with arabionose/melezitose fermentation analysis clearly differentiated only the collection strains into B. longum ssp. longum and B. longum ssp. infantis at the subspecies level. On the other hand, ARDRA analysis differentiated the strains into the B. longum/infantis subspecies using the cleavage analysis of genus-specific amplicon with just one enzyme, Sau3AI. According to our results the majority of the strains belong to the B. longum ssp. infantis (75%). Therefore we suggest ARDRA using Sau3AI restriction enzyme as the first method of choice for distinguishing between B. longum ssp. longum and B. longum ssp. infantis.

• MeSH
• Bifidobacterium chemie   klasifikace   genetika   MeSH
• DNA bakterií chemie   izolace a purifikace   MeSH
• dospělí MeSH
• feces mikrobiologie   MeSH
• fylogeneze MeSH
• kojenec MeSH
• kojení MeSH
• lidé MeSH
• molekulární typizace metody   MeSH
• restrikční mapování MeSH
• RNA ribozomální 16S genetika   MeSH
• shluková analýza MeSH
• technika náhodné amplifikace polymorfní DNA MeSH
• Check Tag
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• srovnávací studie MeSH

Magnetic microspheres P(HEMA-co-EDMA) were used for PCR-ready phage DNA isolation from lysogenic strains of Staphylococcus aureus, including two new clinical isolates. The conditions of phage particle lysis were optimized. The quality of eluted phage DNA was evaluated by PCR. It was demonstrated that PCR-ready phage DNA can be isolated from small volumes of phage lysates (150 microl) by magnetic microspheres. The reported method is very expeditious without using toxic compounds such as phenol or chloroform. It can be used for phage identification and phage gene detection.

• MeSH
• DNA virů genetika   izolace a purifikace   MeSH
• extrakce na pevné fázi metody   MeSH
• financování organizované MeSH
• genetické techniky MeSH
• magnetismus MeSH
• mikrosféry MeSH
• polymerázová řetězová reakce MeSH
• stafylokokové bakteriofágy genetika   izolace a purifikace   MeSH
• Staphylococcus aureus virologie   MeSH

• Publikační typ
• abstrakt z konference MeSH

• Publikační typ
• abstrakty MeSH