A total of, 78 Clostridium septicum (CLSE) isolates were screened for genes encoding: α-toxin, flagellin, and resistance to vancomycin (VANg). The isolates were also tested for their ability to form biofilm and their antibiotic susceptibility. All isolates were positive for α-toxin and flagellin genes. However, only 19 isolates (24.3%) showed prevalence for VANg. We observed the strongest capacity to form a biofilm (100%) in isolates from patients with oncologic or septic and febrile diagnoses. This percentage was also very high in patients with colitis and gastrointestinal hemorrhage (72.7%). No less than 43 isolates showed antibiotic resistance, and 21 were multidrug-resistant (MDR). Interestingly, our studies showed a correlation between antibiotic resistance and biofilm formation. A statistically significant difference was observed between biofilm-forming MDR isolates and those with low/no biofilm-forming ability. However, the most impressive observation was the correlation with mortality rate. While the overall mortality rate for CLSE infections was 16.7% (13/78), the mortality rate for patients infected with MDR isolates forming biofilm moderately or strongly reached 38.1% (8/21). This number increased even further when only infections with the biofilm-forming VANg-positive isolates were considered (61.5%; 8/13). Therefore, the ability of a VANg-positive CLSE isolate to form a biofilm has been suggested as a biomarker of poor prognosis.

• MeSH
• antibakteriální látky * farmakologie   MeSH
• biofilmy MeSH
• Clostridium septicum * MeSH
• flagelin MeSH
• lidé MeSH
• mnohočetná bakteriální léková rezistence genetika   MeSH
• prognóza MeSH
• vankomycin farmakologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

OBJECTIVE: The motility and genotype of the flagellin fliC and fliD genes were investigated in 82 Clostridioides difficile isolates belonging to the ribotypes (RTs): 027 (n = 41), 176 (n = 17), 023 (n = 8), 017 (n = 6) and 046 (n = 10). The reference C. difficile strains 630 and M120 were included as controls for the motility assay. METHODS: A Multiple Locus Variable-number Tandem Repeat Analysis (MLVA) was used to exclude the genetic relatedness of C. difficile isolates belonging to the same RT. The variability of the fliC and fliD genes was determined by PCR-restriction fragment length polymorphism (RFLP) analysis and Sanger sequencing. The motility assay was carried out with 0.175% BHI agar tubes and BHI solid media plates with 0.4% agar. RESULTS: The highest motility was observed in C. difficile RT023 isolates (p < 0.01), followed by RTs 027 and 176. C. difficile isolates of RTs 017 and 046 were less motile than RTs 027, 176 and 023 (p < 0.01). The fliC and fliD genes were present in all clinical isolates irrespective of the motility results. In the fliC gene analysis, four different RFLP groups were identified (I, II, VII, X). The fliC group VII was identified in two RTs (027 and 176), whereas the remaining three groups (I, II and X) belonged to a single RT 046, 017 and 023, respectively. The fliD gene analysis identified four new RFLP groups (a, b, c and d). CONCLUSIONS: C. difficile RT023 is highly motile and its motility is comparable to the hypervirulent RT027 and its genetic relative RT176.

• MeSH
• bakteriální proteiny genetika   MeSH
• Clostridioides difficile * genetika   MeSH
• Clostridioides MeSH
• flagelin genetika   MeSH
• genotyp MeSH
• klostridiové infekce * MeSH
• lidé MeSH
• ribotypizace MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Turkeys and broilers have been identified as important reservoirs for Campylobacter jejuni which is of public health significance. The evaluation of the genotypes among C. jejuni strains within different reservoirs is critical for our understanding of the epidemiology of this infectious agent. The present study aimed to compare the genetic diversity and differences of C. jejuni isolates from turkeys and broilers using flagellin PCR-RFLP typing (flaA typing) technique, in terms of the ease of use and discriminatory power. Sixty C. jejuni isolates were detected biochemically and confirmed by duplex-PCR from turkeys and broilers (30 strains from each bird species). Then, a flaA gene fragment (1725 bp) of C. jejuni isolates was amplified and amplicons were digested with HpyF3I enzyme. Restriction analysis by HpyF3I gave four different flaA patterns (H1, H2, H3, H4) among all tested C. jejuni isolates. In broiler isolates, all four patterns were observed but in turkey isolates, only H2 and H4 patterns were present. The results clearly demonstrated that distribution of the flaA typing patterns differed depending on the host species (broiler/turkey). H1 and H3 flaA types are more prevalent in broiler than turkey isolates, while H2 type is significantly more prevalent within isolates from turkey (p < 0.05). The flaA typing technique by digestion with HpyF3I enzyme can almost give us a clue to the source of infection in local outbreaks.

• MeSH
• Campylobacter jejuni klasifikace   genetika   MeSH
• DNA bakterií genetika   MeSH
• flagelin genetika   MeSH
• genetická variace MeSH
• kampylobakterové infekce mikrobiologie   veterinární   MeSH
• krocani MeSH
• kur domácí MeSH
• nemoci drůbeže mikrobiologie   MeSH
• restrikční mapování veterinární   MeSH
• techniky typizace bakterií veterinární   MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Recognition of pathogen-associated molecular patterns (PAMPs) is crucial for plant defence against pathogen attack. The best characterized PAMP is flg22, a 22 amino acid conserved peptide from flagellin protein. In Arabidopsis thaliana, flg22 is recognized by the flagellin sensing 2 (FLS2) receptor. In this study, we focused on biotic stress responses triggered by flg22 after exposure to temporary heat stress (HS). It is important to study the reactions of plants to multiple stress conditions because plants are often exposed simultaneously to a combination of both abiotic and biotic stresses. Transient early production of reactive oxygen species (ROS) is a well-characterized response to PAMP recognition. We demonstrate the strong reduction of flg22-induced ROS production in A. thaliana after HS treatment. In addition, a decrease in FLS2 transcription and a decrease of the FLS2 presence at the plasma membrane are shown after HS. In summary, our data show the strong inhibitory effect of HS on flg22-triggered events in A. thaliana. Subsequently, temporary HS strongly decreases the resistance of A. thaliana to Pseudomonas syringae. We propose that short exposure to high temperature is a crucial abiotic stress factor that suppresses PAMP-triggered immunity, which subsequently leads to the higher susceptibility of plants to pathogens.

• MeSH
• alarminy metabolismus   MeSH
• Arabidopsis účinky léků   genetika   imunologie   mikrobiologie   MeSH
• flagelin farmakologie   MeSH
• genetická transkripce účinky léků   MeSH
• imunita rostlin * účinky léků   MeSH
• nemoci rostlin imunologie   mikrobiologie   MeSH
• odolnost vůči nemocem imunologie   MeSH
• proteiny huseníčku genetika   metabolismus   MeSH
• Pseudomonas syringae účinky léků   fyziologie   MeSH
• reakce na tepelný šok * účinky léků   MeSH
• regulace genové exprese u rostlin účinky léků   MeSH
• respirační vzplanutí účinky léků   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Proinflammatory cytokines play important roles in the pathogenesis of diseases caused by enterohemorrhagic Escherichia coli (EHEC) O157, but the spectrum of bacterial components involved in the proinflammatory responses is not fully understood. Here, we investigated the abilities of outer membrane vesicles (OMVs), nanoparticles released by EHEC O157 during growth, to induce production of proinflammatory cytokines in human intestinal epithelial cells. OMVs from both EHEC O157:H7 and sorbitol-fermenting (SF) EHEC O157:H- induced production of interleukin-8 (IL-8) in Caco-2, HCT-8, and HT-29 intestinal epithelial cell lines. H7 flagellin was the key IL-8-inducing component of EHEC O157:H7 OMVs, whereas cytolethal distending toxin V and O157 lipopolysaccharide (LPS) largely contributed to IL-8 production elicited by flagellin-lacking OMVs from SF EHEC O157:H-. The H7 flagellin-mediated signaling via Toll-like receptor (TLR) 5, and O157 LPS-mediated signaling via TLR4/MD-2 complex, which were followed by activation of the nuclear factor NF-κB were major pathways underlying IL-8 production induced by EHEC O157 OMVs. The proinflammatory and immunomodulatory capacities of EHEC O157 OMVs have pathogenetic implications and support the OMVs as suitable vaccine candidates.

• MeSH
• buněčná membrána metabolismus   MeSH
• buňky HT-29 MeSH
• Caco-2 buňky MeSH
• epitelové buňky metabolismus   MeSH
• Escherichia coli O157 patogenita   MeSH
• faktory virulence metabolismus   MeSH
• flagelin metabolismus   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   patologie   MeSH
• interleukin-8 biosyntéza   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• NF-kappa B metabolismus   MeSH
• proteiny vnější bakteriální membrány metabolismus   MeSH
• proteiny z Escherichia coli metabolismus   MeSH
• signální transdukce MeSH
• střevní sliznice cytologie   mikrobiologie   patologie   MeSH
• toll-like receptor 4 metabolismus   MeSH
• toll-like receptor 5 metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Biogenesis of the plant secondary cell wall involves many important aspects, such as phenolic compound deposition and often silica encrustation. Previously, we demonstrated the importance of the exocyst subunit EXO70H4 for biogenesis of the trichome secondary cell wall, namely for deposition of the autofluorescent and callose-rich cell wall layer. Here, we reveal that EXO70H4-driven cell wall biogenesis is constitutively active in the mature trichome, but also can be activated elsewhere upon pathogen attack, giving this study a broader significance with an overlap into phytopathology. To address the specificity of EXO70H4 among the EXO70 family, we complemented the exo70H4-1 mutant by 18 different Arabidopsis (Arabidopsis thaliana) EXO70 paralogs subcloned under the EXO70H4 promoter. Only EXO70H4 had the capacity to rescue the exo70H4-1 trichome phenotype. Callose deposition phenotype of exo70H4-1 mutant is caused by impaired secretion of PMR4, a callose synthase responsible for the synthesis of callose in the trichome. PMR4 colocalizes with EXO70H4 on plasma membrane microdomains that do not develop in the exo70H4-1 mutant. Using energy-dispersive x-ray microanalysis, we show that both EXO70H4- and PMR4-dependent callose deposition in the trichome are essential for cell wall silicification.

• MeSH
• Arabidopsis účinky léků   genetika   metabolismus   MeSH
• buněčná membrána účinky léků   metabolismus   MeSH
• buněčná stěna účinky léků   metabolismus   MeSH
• epidermis rostlin cytologie   účinky léků   metabolismus   MeSH
• fenotyp MeSH
• flagelin farmakologie   MeSH
• glukany MeSH
• glukosyltransferasy metabolismus   MeSH
• mutace genetika   MeSH
• oxid křemičitý metabolismus   MeSH
• podjednotky proteinů chemie   metabolismus   MeSH
• proteinové domény MeSH
• proteiny huseníčku chemie   metabolismus   MeSH
• regulace genové exprese u rostlin účinky léků   MeSH
• trichomy metabolismus   MeSH
• upregulace účinky léků   MeSH
• vezikulární transportní proteiny chemie   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: Microbial-associated molecular patterns activate several MAP kinases, which are major regulators of the innate immune response in Arabidopsis thaliana that induce large-scale changes in gene expression. Here, we determine whether microbial-associated molecular pattern-triggered gene expression involves modifications at the chromatin level. RESULTS: Histone acetylation and deacetylation are major regulators of microbial-associated molecular pattern-triggered gene expression and implicate the histone deacetylase HD2B in the reprogramming of defence gene expression and innate immunity. The MAP kinase MPK3 directly interacts with and phosphorylates HD2B, thereby regulating the intra-nuclear compartmentalization and function of the histone deacetylase. CONCLUSIONS: By studying a number of gene loci that undergo microbial-associated molecular pattern-dependent activation or repression, our data reveal a mechanistic model for how protein kinase signaling directly impacts chromatin reprogramming in plant defense.

• MeSH
• Arabidopsis imunologie   MeSH
• chromatin fyziologie   MeSH
• flagelin imunologie   MeSH
• fosforylace MeSH
• fyziologický stres MeSH
• histondeacetylasy metabolismus   MeSH
• histony metabolismus   MeSH
• imunita rostlin * MeSH
• mitogenem aktivované proteinkinasy kinas metabolismus   MeSH
• přirozená imunita MeSH
• proteiny huseníčku metabolismus   MeSH
• restrukturace chromatinu * MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Membrane microdomains play vital roles in the process of bacterial infection. The membrane microdomain-associated protein Flot1 acts in an endocytic pathway and is required for seedling development, however, whether Flot1 is a part of host defense mechanisms remains unknown. During an analysis of callose deposition, we found that Flot1 amiRNAi mutants exhibited defects in response to flg22. Using variable-angle total internal reflection fluorescence microscopy (VA-TIRFM), structured illumination microscopy (SIM) and fluorescence cross spectroscopy (FCS), we determined that the dynamic behavior of GFP-Flot1 in Arabidopsis thaliana cotyledon epidermal cells changed significantly in plants treated with the elicitor flg22. Moreover, we found that Flot1 was constitutively recycled via an endocytic pathway and that flg22 could promote endocytosis. Importantly, targeting of Flot1 to the late endosome/vacuole for degradation increased in response to flg22 treatment; immunoblot analysis showed that when triggered by flg22, GFP-Flot1 was gradually degraded in a time-dependent manner. Taken together, these findings support the hypothesis that the changing of dynamics and oligomeric states can promote the endocytosis and degradation of Flot1 under flg22 treatment in plant cells.

• MeSH
• Arabidopsis účinky léků   genetika   metabolismus   MeSH
• endocytóza účinky léků   genetika   fyziologie   MeSH
• flagelin farmakologie   MeSH
• fluorescenční mikroskopie MeSH
• geneticky modifikované rostliny genetika   metabolismus   MeSH
• kotyledon genetika   metabolismus   MeSH
• proteiny huseníčku genetika   metabolismus   MeSH
• regulace genové exprese u rostlin účinky léků   genetika   MeSH
• Publikační typ
• časopisecké články MeSH

PURPOSE: For simultaneous detection of Borrelia miyamotoi (relapsing fever spirochete) and Borrelia burgdorferi sensu lato, we have developed a duplex real-time PCR targeting the flagellin gene (flaB; p41), a locus frequently used in routine diagnostic PCR for B. burgdorferi s.l. detection. METHODS: Primers and probes were designed using multiple alignments of flaB sequences of B. miyamotoi and B. burgdorferi s.l. species. The sensitivity and specificity of primers and probes were determined using serial dilutions (ranging from 10(4) to 10(-1)) of B. miyamotoi and B. burgdorferi s.l. DNA and of several species of relapsing fever spirochetes. Conventional PCR on recG and glpQ and sequencing of p41 PCR products were used to confirm the species assignment. RESULTS: The detection limit of both singleplex and duplex PCR was 10 genome equivalents except for B. spielmanii and two B. garinii genotypes which showed a detection limit of 10(2) genome equivalents. There was no cross reactivity of the B. miyamotoi primers/probes with B. burgdorferi s.l. DNA, while the B. burgdorferi s.l. primer/probe generated a signal with B. hermsii DNA. Out of 2341 Ixodes ricinus ticks from Germany and Slovakia that were screened simultaneously for the presence of B. miyamotoi and B. burgdorferi s.l., 52 were positive for B. miyamotoi and 276 for B. burgdorferi s.l., denoting an average prevalence of 2.2% for B. miyamotoi and 11.8% for B. burgdorferi s.l., and B. miyamotoi DNA was also detectable by PCR using artificial clinical samples. CONCLUSION: The duplex real-time PCR developed here represents a method that permits simultaneous detection and differentiation of B. burgdorferi s.l. and B. miyamotoi in environmental and potentially clinical samples.

• MeSH
• Borrelia klasifikace   genetika   MeSH
• DNA primery genetika   MeSH
• flagelin genetika   MeSH
• klíště mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• lidé MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• oligonukleotidové sondy genetika   MeSH
• senzitivita a specificita MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• hodnotící studie MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Německo MeSH
• Slovenská republika MeSH

Two different genetic loci, flaB and ospC, were employed to assign genospecies and OspC phylogenetic type to 18 strains isolated from ticks collected in Pisárky, a suburban park in the city of Brno, Czech Republic. The RFLP analysis revealed three different genospecies (B. afzelii, B. garinii, and B. valaisiana). Three samples from the collection contained more than one genospecies. In the other 15 strains, nucleotide sequences of flaB and ospC were determined. The following phylogenetic analysis assigned 12 isolates to genospecies B. garinii and three to B. afzelii. These isolates were further subdivided into seven distinct ospC groups. The most related OspC types were G2, G4, and G5 (B. garinii) and A3 and A8 (B. afzelii).

• MeSH
• antigeny bakteriální genetika   MeSH
• Borrelia burgdorferi komplex klasifikace   MeSH
• DNA bakterií genetika   MeSH
• flagelin genetika   MeSH
• fylogeneze MeSH
• klíště mikrobiologie   MeSH
• lymeská nemoc MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• proteiny vnější bakteriální membrány genetika   MeSH
• sérotypizace * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Česká republika MeSH