Legionella pneumophila, the causative agent of Legionnaires' disease, is a facultative intracellular pathogen that survives inside phagocytic host cells by establishing a protected replication niche, termed the "-containing vacuole" (LCV). To form an LCV and subvert pivotal host pathways, L. pneumophila employs a type IV secretion system (T4SS), which translocates more than 300 different effector proteins into the host cell.
View Article and Find Full Text PDFThe swimming larvae of many marine animals identify a location on the sea floor to undergo metamorphosis based on the presence of specific bacteria. Although this microbe-animal interaction is critical for the life cycles of diverse marine animals, what types of biochemical cues from bacteria that induce metamorphosis has been a mystery. Metamorphosis of larvae of the tubeworm is induced by arrays of phage tail-like contractile injection systems, which are released by the bacterium .
View Article and Find Full Text PDFElectron cryotomography is able to visualize macromolecular complexes in their cellular context, in a frozen-hydrated state, and in three dimensions. The method, however, is limited to relatively thin samples. Cryo-focused ion beam (FIB) milling is emerging as a powerful technique for sample thinning prior to cryotomography imaging.
View Article and Find Full Text PDFBacterium-host interactions are important for diverse ecological settings including pathogenicity and symbiosis. Electron cryotomography is a powerful method for studying the macromolecular complexes that mediate such interactions in situ. The main limitation of electron cryotomography is its restriction to relatively thin samples such as individual bacterial cells.
View Article and Find Full Text PDFContractile injection systems mediate bacterial cell-cell interactions by a bacteriophage tail-like structure. In contrast to extracellular systems, the type 6 secretion system (T6SS) is defined by intracellular localization and attachment to the cytoplasmic membrane. Here we used cryo-focused ion beam milling, electron cryotomography, and functional assays to study a T6SS in The in situ architecture revealed three modules, including a contractile sheath-tube, a baseplate, and an anchor.
View Article and Find Full Text PDFMethods Mol Biol
March 2018
The unique property of electron cryotomography (ECT) is its capability to resolve the structure of macromolecular machines in their cellular context. The integration of ECT data with high-resolution structures of purified subcomplexes and live-cell fluorescence light microscopy can generate pseudo-atomic models that lead to a mechanistic understanding across size and time scales. Recent advances in electron detection, sample thinning, data acquisition, and data processing have significantly enhanced the applicability and performance of ECT.
View Article and Find Full Text PDFDiverse animal taxa metamorphose between larval and juvenile phases in response to bacteria. Although bacteria-induced metamorphosis is widespread among metazoans, little is known about the molecular changes that occur in the animal upon stimulation by bacteria. Larvae of the tubeworm Hydroides elegans metamorphose in response to surface-bound Pseudoalteromonas luteoviolacea bacteria, producing ordered arrays of phage tail-like metamorphosis-associated contractile structures (MACs).
View Article and Find Full Text PDFSEM analysis of endodontic instruments from a Ni-Ti rotary system was assessed, before and after using them, considering their defects and deformations. Twenty Twisted File®, BioRąCe®, Mtwo®, and EndoWave® instruments were micrographed at 190× magnification. The files were washed and micrographed again to view alterations as to the presence or absence of irregular edges, grooves, microcavities, and scraping.
View Article and Find Full Text PDFPhotomed Laser Surg
December 2011
Background And Objective: Photodynamic therapy (PDT) is a new technique introduced in endodontics that combines the action of a photosensitizer (dye) and a low intensity light source. Currently, there are no PDT studies evaluating the microbial disinfection of root canals in order to compare the effects of light delivery systems in the photosensitizer activation. The aim of this study was to evaluate the PDT effectiveness in reducing Enterococcus faecalis, with and without the aid of an intracanal optical fiber.
View Article and Find Full Text PDFBackground: The TolC protein from Sinorhizobium meliloti has previously been demonstrated to be required for establishing successful biological nitrogen fixation symbiosis with Medicago sativa. It is also needed in protein and exopolysaccharide secretion and for protection against osmotic and oxidative stresses. Here, the transcriptional profile of free-living S.
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