Specific quantification of root-colonizing arbuscular mycorrhizal fungi (AMF) by quantitative real-time PCR is a high-throughput technique, most suitable for determining abundances of AMF species or isolates in previously characterized experimental systems. The principal steps are the choice and validation of an appropriate assay to specifically amplify a gene fragment of the target AMF, preparation of templates from root samples, and quantification of the fungal gene copy numbers in these templates. The use of a suitable assay is crucial for a correct data collection but also highly specific for each experimental system and is therefore covered by general recommendations. Subsequently, specific steps are described for the validation of the assay using a standard dilution series, the determination of appropriate dilutions of DNA extracts from roots, and the quantification of the gene copy numbers in samples including calculations.

• MeSH
• DNA fungální genetika   izolace a purifikace   MeSH
• genová dávka genetika   MeSH
• kořeny rostlin genetika   mikrobiologie   MeSH
• kvantitativní polymerázová řetězová reakce metody   MeSH
• mykorhiza genetika   izolace a purifikace   MeSH
• půda MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Stem cell regulation plays a crucial role during development and homeostasis. Here, an essential source of Wnts from Gli1+ stem/progenitor cells was identified in the murine molar. Loss of Wnt production in Gli1+ apical stem/progenitor cells led to loss of Axin2 at the root apex, mis-regulation of SOX9, loss of BMP and Hh signaling, and truncation of root development. In the absence of Wnt signals, the root epithelium lost its integrity and epithelial identity. This phenotype could be partially mimicked by loss of Sox9 in the Gli1 population. Stabilization of Wnt signaling in the apical papilla led to rapid unordered differentiation of hard tissues and fragmentation of the epithelial root sheath. Wnt signaling from Gli1+ stem/progenitor cells, therefore, orchestrates root development, coordinating mesenchymal and epithelial interactions via SOX9 to regulate stem/progenitor cell expansion and differentiation. Our results demonstrate that disparate stem/progenitor cell populations are unified in their fundamental signaling interactions.

• MeSH
• buněčná diferenciace genetika   MeSH
• kmenové buňky * metabolismus   MeSH
• myši MeSH
• protein Gli1 genetika   metabolismus   MeSH
• signální dráha Wnt * MeSH
• zubní kořen metabolismus   MeSH
• zvířata MeSH
• Check Tag
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Spatial distribution of ectomycorrhizae-associated basidiomycetes was determined in oakbirch forest using terminal restriction fragment length polymorphism (T-RFLP) analysis. The data were correlated with actual soil humidity, pH, electric conductivity of the soil extract, absorbance A(465) and A(665) of water and alkali soil extracts and with the ratio A(465)/A(665) (parameter A4/A6). Natural non-homogeneity of the soil parameters was used as experimental gradient. Distance-based redundancy analysis of the T-RFLP data (with soil parameters being taken as environmental parameters) provided significant results when ITS1F-terminanted restriction fragments were analyzed. Among other fungi, a Mycena galericulata related fungus was observed to correlate negatively with A4/A6, indicating its association with highly humified soil organic matter. Positive association of other, unidentified fungi with A4/A6 was also observed. Several other unidentified fungi negatively correlated with electric conductivity of the soil extract. The results may explain nonhomogeneity of the spatial distribution of the fungi associated with ectomycorrhizae as a result of their interaction with non-homogeneous soil environment.

• MeSH
• Agaricales genetika   izolace a purifikace   růst a vývoj   MeSH
• Basidiomycota genetika   izolace a purifikace   klasifikace   růst a vývoj   MeSH
• bříza mikrobiologie   růst a vývoj   MeSH
• dub (rod) mikrobiologie   růst a vývoj   MeSH
• elektrolyty analýza   MeSH
• financování organizované MeSH
• kořeny rostlin mikrobiologie   MeSH
• mykorhiza MeSH
• organické látky analýza   MeSH
• polymorfismus délky restrikčních fragmentů MeSH
• půda MeSH
• půdní mikrobiologie MeSH
• stromy mikrobiologie   růst a vývoj   MeSH
• Geografické názvy
• Česká republika MeSH

Zalomení nástroje v kořenovém kanálku patří mezi komplikace endodontické terapie. K odstranění fragmentu nástroje z kořenového kanálku ortográdním přístupem bylo vyvinuto několik metod. Úspěšné odstranění fragmentu nástroje však i přesto zůstává velmi komplikovaným výkonem, který je plně vázán na použití ultrazvukových přístrojů a práci pod operačním mikroskopem. Úspěšnost odstranění fragmentu kořenového nástroje se významně zvyšuje s erudicí operatéra a délkou jeho praxe s operačním mikroskopem. Pohybuje se kolem 90 %.

Fracture of an endodontic file is to be seen as a complication of the root canal treatment. Several protocols have been developed for broken endodontics files removal in the orthograde or nonsurgical approach. Nevertheless, a successful removal of a broken file fragment remains to be a very complicated procedure making the use of ultrasonic devices and a microscope necessary. The success rate of broken endodontic file removal is rising significantly according to the operator´s experience in the use of a microscope and can mount up to 90 %.

• MeSH
• peroperační komplikace prevence a kontrola   terapie   MeSH
• stomatochirurgické výkony metody   využití   MeSH
• terapie kořenového kanálku MeSH
• zubní nástroje MeSH

Based on molecular markers (COII and ITS1-ITS2) and morphological data, we describe three new Neotropical species of Gyrodactylus von Nordmann, 1832 from Scleromystax barbatus (Quoy et Gaimard) and Scleromystax macropterus (Regan) from southern Brazil. The three new species can be distinguished from each other by sequences of both molecular markers and morphology of hooks and anchors. Gyrodactylus bueni sp. n. is characterised by having hook with shaft curved, heel straight, shelf straight, toe pointed, anchor with superficial root slender, elongate and male copulatory organ armed with two rows of spinelets. Gyrodactylus major sp. n. presents hook with shaft, point curved, proximal shaft straight, heel convex, shelf convex, toe concave, anchor with superficial root robust and male copulatory organ armed with two rows of spinelets. Gyrodactylus scleromystaci sp. n. presents hook with shaft, point recurved, heel convex, shelf convex, toe pointed, anchor with superficial root curved and male copulatory organ armed with two rows of spinelets. These species appear to be closely related to other species of Gyrodactylus known from other species of Callichthyidae. These new species, however, differ by the comparative morphology of the haptoral hard structures and molecular data. Comparative analysis of sequences from these species of Gyrodactylus suggests that the COII gene may represent an important marker for the taxonomy of species of Gyrodactylidae and, perhaps, for species of other lineages of Monogenoidea.

• MeSH
• druhová specificita MeSH
• genetické markery * MeSH
• infekce červy třídy Trematoda parazitologie   veterinární   MeSH
• nemoci ryb parazitologie   MeSH
• respirační komplex IV genetika   metabolismus   MeSH
• ryby MeSH
• taxonomické DNA čárové kódování MeSH
• Trematoda klasifikace   enzymologie   genetika   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

File breakage is a reality of life. If anyone says, he doesn't separate the tip of file in the canal, don't trust him. He doesn't do much endo. OK, file broke in the canal. What should we do? In this article author sumarises the factors, which are needed to be taken into the consideration, while trying to solve “separated file” case. In the first place anatomy of root canal, location of the fragment, vitality of the pulp, type of broken instrument. All these information show us the way we can follow. Of course, having adequate skills together with adequate tools, including magnification and ultrasonic.

• MeSH
• cizí tělesa * etiologie   chirurgie   radiografie   MeSH
• dospělí MeSH
• lidé MeSH
• preparace kořenového kanálku * metody   přístrojové vybavení   MeSH
• prognóza MeSH
• selhání zařízení * MeSH
• terapie kořenového kanálku metody   přístrojové vybavení   MeSH
• ultrazvuková chirurgie metody   přístrojové vybavení   MeSH
• výsledek terapie MeSH
• Check Tag
• dospělí MeSH
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• kazuistiky MeSH

The complicated crown-root fracture of young permanent teeth is an uncommon traumatic dental injury that is usually treated in a complex way and is demanding not only for the dentist but even for the treated child. In this case report, we present the conservative treatment of a maxillary central incisor in a 10-year-old boy after a traumatic dental injury. Treatment included partial pulpotomy and adhesive fragment reattachment after reflection of the mucoperiosteal flap. The patient was fully asymptomatic at 24-month follow-up, with an aesthetically acceptable outcome. Vital pulp therapy and adhesive fragment reattachment can be a viable treatment option for complicated crown-root fractures, especially when treating immature permanent teeth.

• Publikační typ
• kazuistiky MeSH

Klinické zkušenosti z let 1953-58 v ošetřování bederního a krčního syndromu diskogenní komprese kaudy a míchy, spojené s výsledky výzkumu jeho patogeneze, terapie a prevence. Práce vymezuje kritéria pro indikaci chirurgické léčby a rozebírá její výsledky.

• MeSH
• neurochirurgické výkony MeSH
• radikulopatie chirurgie   MeSH
• výhřez meziobratlové ploténky chirurgie   MeSH

• Konspekt
• Ortopedie. Chirurgie. Oftalmologie
• NLK Obory
• chirurgie
• neurochirurgie
• NLK Publikační typ
• studie

As part of the EU project MULTIBARRIERS, six new endogenous aerobic bacterial isolates able to grow in the presence of BTmX (benzene, toluene, m-xylene) were characterized with respect to their growth specificities. Preliminary analysis included restriction fragment length polymorphism profiles and 16S rDNA sequencing. The diversity of these strains was confirmed by denaturing gradient gel electrophoresis. Additional aerobic bacterial strains were isolated from the rhizospheres of plants grown in polychlorinated biphenyl (PCB)-contaminated soils. Pot experiments were designed to show the beneficial effect of plants on the bacterial degradation of PCBs. The effect of PCB removal from soil was evaluated and bacteria isolated from three different plant species were examined for the presence of the bph operon.

• MeSH
• Actinobacteria genetika   klasifikace   metabolismus   MeSH
• aromatické uhlovodíky metabolismus   MeSH
• biodegradace MeSH
• chemické látky znečišťující vodu metabolismus   MeSH
• dekontaminace metody   MeSH
• ekosystém MeSH
• financování organizované MeSH
• kořeny rostlin mikrobiologie   MeSH
• látky znečišťující půdu metabolismus   MeSH
• Proteobacteria genetika   klasifikace   metabolismus   MeSH
• půdní mikrobiologie MeSH
• rostliny mikrobiologie   MeSH

Miscanthus x giganteus is suggested as a good candidate for phytostabilization of metal-polluted soils. Its late harvest in winter generates large amounts of leaf litter on the soil surface. However, little is known about the mobility and the bioavailability of metals following leaf decomposition and the consequences on the succeeding culture. Ex situ artificial aging for 1, 3, and 6 months was conducted with miscanthus leaf fragments incorporated into three agricultural soils displaying a gradient concentration in Cd (0.6, 3.1 and 7.9 mg kg-1), Pb (32.0, 194.6 and 468.6 mg kg-1), and Zn (48.4, 276.3 and 490.2 mg kg-1) to simulate the leaf litter input over 20 years of miscanthus culture. We investigated the impacts on physicochemical and biological soil parameters, CaCl2-extractable metal, and their subsequent ryegrass shoot concentrations, and hence on ryegrass health. The results showed that the amended soils possessed higher pH along with greater available phosphorous and soil organic carbon values. The respiratory activity and microbial biomass carbon in the amended soils increased mainly after 1 month of aging, and decreased afterwards. Despite the higher Pb- and Zn-CaCl2 extractability in the amended soils, the phytoavailability slightly increased only in the most contaminated soils. Moreover, leaf incorporation did not affect the ryegrass biomass, photosynthetic pigment contents, nor the antioxidative enzyme activities. Conclusively, leaf incorporation induced slight variations in soil physicochemical and biological parameters, as well as metal extractability, but not to an extent that might cause a considerable threat to the subsequent culture. Nevertheless, these results are preliminary data that require confirmation by long-term in-situ experimentations as they reflect the modelization of long-term impact of leaf decomposition on soil-plant system.

• MeSH
• biodegradace MeSH
• biologická dostupnost MeSH
• biomasa MeSH
• jílek růst a vývoj   metabolismus   MeSH
• kořeny rostlin metabolismus   MeSH
• látky znečišťující půdu analýza   metabolismus   MeSH
• lipnicovité * růst a vývoj   metabolismus   MeSH
• listy rostlin metabolismus   MeSH
• půda chemie   MeSH
• teoretické modely * MeSH
• těžké kovy analýza   metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH