Genetic and Structural Analyses of Cytoplasmic Filaments of Wild-Type Treponema phagedenis and a Flagellar Filament-Deficient Mutant

Unique cytoplasmic filaments are found in the treponeme genus of spirochete bacteria. Their function is unknown, but their location underneath the periplasmic flagellar filaments (PFF) suggests a role in motility and/or cell structure. To better understand these unique structures, the gene coding for the cytoplasmic filaments, cfpA, was identified in various treponemal species. Treponema phagedenis cfpA was 2,037 nucleotides long, and the encoded polypeptide showed 78 to 100% amino acid sequence identity with the partial sequence of CfpA from T. denticola, T. vincentii, and T. pallidum subsp. pertenue. Wild-type T. phagedenis and a PFF-deficient isolate were analyzed by electron microscopy to assess the structural relationship of the cytoplasmic filaments and the PFF. The number of cytoplasmic filaments per cell of T. phagedenis (mean, 5.7) was compared with the number of PFF at each end of the cell (mean, 4.7); the results suggest that there is no direct one-to-one correlation at the cell end. Moreover, a structural link between these structures could not be demonstrated. The cytoplasmic filaments were also analyzed by electron microscopy at different stages of cell growth; this analysis revealed that they are cleaved before or during septum formation and before the nascent formation of PFF. A PFF-deficient mutant of T. phagedenis possessed cytoplasmic filaments similar to those of the wild type, suggesting that intact PFF are not required for their assembly and regulation. The extensive conservation of CfpA among pathogenic spirochetes suggests an important function, and structural analysis suggests that it is unlikely that the cytoplasmic filaments and the flagellar apparatus are physically linked

Native cellular architecture of Treponema denticola revealed by cryo-electron tomography

Using cryo-electron tomography, we are developing a refined description of native cellular structures in the pathogenic spirochete Treponema denticola. Tightly organized bundles of periplasmic flagella were readily observed in intact plunge-frozen cells. The periplasmic space was measured in both wild-type and aflagellate strains, and found to widen by less than the diameter of flagella when the latter are present. This suggests that a structural change occurs in the peptidoglycan layer to accommodate the presence of the flagella. In dividing cells, the flagellar filaments were found to bridge the cytoplasmic cylinder constriction site. Cytoplasmic filaments, adjacent to the inner membrane, run parallel to the tightly organized flagellar filaments. The cytoplasmic filaments may be anchored by a narrow plate-like structure. The tapering of the cell ends was conserved between cells, with a patella-shaped structure observed in the periplasm at the tip of each cytoplasmic cylinder. Several incompletely characterized structures have been observed in the periplasm between dividing cells, including a cable-like structure linking two cytoplasmic cylinders and complex foil-shaped structures

Tomographic reconstruction of treponemal cytoplasmic filaments reveals novel bridging and anchoring components

An understanding of the involvement of bacterial cytoplasmic filaments in cell division requires the elucidation of the structural organization of those filamentous structures. Treponemal cytoplasmic filaments are composed of one protein, CfpA, and have been demonstrated to be involved in cell division. In this study, we used electron tomography to show that the filaments are part of a complex with a novel molecular organization that includes at least two distinct features decorating the filaments. One set of components appears to anchor the filaments to the cytoplasmic membrane. The other set of components appears to bridge the cytoplasmic filaments on the cytoplasmic side, and to be involved in the interfilament spacing within the cell. The filaments occupy between 3 and 18% of the inner surface of the cytoplasmic membrane. These results reveal a novel filamentous molecular organization of independent filaments linked by bridges and continuously anchored to the membrane

Semiconductor-enriched single wall carbon nanotube networks applied to field effect transistors

Substantial progress on field effect transistors "FETs" consisting of semiconducting single wall carbon nanotubes "s-SWNTs" without detectable traces of metallic nanotubes and impurities is reported. Nearly perfect removal of metallic nanotubes is confirmed by optical absorption, Raman measurements, and electrical measurements. This outstanding result was made possible in particular by ultracentrifugation (150 000 g) of solutions prepared from SWNT powders using polyfluorene as an extracting agent in toluene. Such s-SWNTs processable solutions were applied to realize FET, embodying randomly or preferentially oriented nanotube networks prepared by spin coating or dielectrophoresis. Devices exhibit stable p-type semiconductor behavior in air with very promising characteristics. The on-off current ratio is 10^5, the on-current level is around 10 $\mu$A, and the estimated hole mobility is larger than 2 cm2 / V s

NextMed, Augmented and Virtual Reality platform for 3D medical imaging visualization

The visualization of the radiological results with more advanced techniques than the current ones, such as Augmented Reality and Virtual Reality technologies, represent a great advance for medical professionals, by eliminating their imagination capacity as an indispensable requirement for the understanding of medical images. The problem is that for its application it is necessary to segment the anatomical areas of interest, and this currently involves the intervention of the human being. The Nextmed project is presented as a complete solution that includes DICOM images import, automatic segmentation of certain anatomical structures, 3D mesh generation of the segmented area, visualization engine with Augmented Reality and Virtual Reality, all thanks to different software platforms that have been implemented and detailed, including results obtained from real patients. We will focus on the visualization platform using both Augmented and Virtual Reality technologies to allow medical professionals to work with 3d model representation of medical images in a different way taking advantage of new technologies

The Unseen World: Environmental Microbial Sequencing and Identification Methods for Ecologists

Microorganisms inhabit almost every environment, comprise the majority of diversity on Earth, are important in biogeochemical cycling, and may be vital to ecosystem responses to large-scale climatic change. In recent years, ecologists have begun to use rapidly advancing molecular techniques to address questions about microbial diversity, biogeography, and responses to environmental change. Studies of microbes in the environment generally focus on three broad objectives: determining which organisms are present, what their functional capabilities are, and which are active at any given time. However, comprehending the range of methodologies currently in use can be daunting. To provide an overview of environmental microbial sequence data collection and analysis approaches, we include case studies of microbiomes ranging from the human mouth to geothermal springs. We also suggest contexts in which each technique can be applied and highlight insights that result from their use

The effect of induced sadness and moderate depression on attention networks

This study investigates how sadness and minor/moderate depression influences the three functions of attention: alerting, orienting, and executive control using the attention network test. The aim of the study is to investigate whether minor to moderate depression is more similar to sadness or clinical depression with regards to attentional processing. It was predicted that both induced sadness and minor to moderate depression will influence executive control by narrowing spatial attention and in turn this will lead to less interference from the flanker items (i.e., less effects of congruency) due to a focused attentional state. No differences were predicted for alerting or orienting functions. The results from the two experiments, the first inducing sadness (Experiment 1) and the second measuring subclinical depression (Experiment 2), show that, as expected, participants who are sad or minor to moderately depressed showed less flanker interference compared to participants who were neither sad nor depressed. This study provides strong evidence, that irrespective of its aetiology, sadness and minor/moderate depression have similar effects on spatial attention

Force and acceleration characteristics of military foot drill: implications for injury risk in recruits

Background: Foot drill involving marching and drill manoeuvres is conducted regularly during basic military recruit training. Characterising the biomechanical loading of foot drill will improve our understanding of the contributory factors to lower limb overuse injuries in recruits. Aim: Quantify and compare forces, loading rates and accelerations of British Army foot drill, within and between trained and untrained personnel. Methods: 24 trained soldiers (12 men and 12 women; TRAINED) and 12 civilian men (UNTRAINED) performed marching and five drill manoeuvres on force platforms; motion capture recorded tibial position. Peak vertical impact force (PF), peak vertical loading rate (PLR), expressed as multiples of body weight (BW) and peak tibial impact acceleration (PTA) were recorded. Results: Drill manoeuvre PF, PLR and PTA were similar, but higher in TRAINED men (PF, PLR: p<0.01; PTA: p<0.05). Peak values in TRAINED men were shown for the halt (mean (SD); PF: 6.5 (1.5) BW; PLR: 983 (333) BW/s PTA; PTA: 207 (57) m/s2) and left turn (PF: 6.6 (1.7) BW; PLR: 928 (300) BW/s; 184 (62) m/s2). Marching PF, PLR, PTA were similar between groups and lower than all drill manoeuvres (PF: 1.1–1.3 BW; PLR: 42–70 BW/s; p<0.01; PTA: 23–38 m/s2; p<0.05). Conclusions: Army foot drill generates higher forces, loading rates and accelerations than activities such as running and load carriage, while marching is comparable to moderate running (10.8 km/h). The large biomechanical loading of foot drill may contribute to the high rate of overuse injuries during initial military training, and strategies to regulate/reduce this loading should be explored

Investigating online training in government agencies: Designing adaptive web-based instructional programmes to reskill the workforce

The government sector relies on continual employee reskilling. This journal paper discusses a three year Australian Research Council (ARC) funded project, which seeks to facilitate cost effective online-learning, using advanced information and communications technology (ICT) tools to enhance workforce training, with assured predictable outcomes

Cryo-Electron Tomography Elucidates the Molecular Architecture of Treponema pallidum, the Syphilis Spirochete

Cryo-electron tomography (CET) was used to examine the native cellular organization of Treponema pallidum, the syphilis spirochete. T. pallidum cells appeared to form flat waves, did not contain an outer coat and, except for bulges over the basal bodies and widening in the vicinity of flagellar filaments, displayed a uniform periplasmic space. Although the outer membrane (OM) generally was smooth in contour, OM extrusions and blebs frequently were observed, highlighting the structure’s fluidity and lack of attachment to underlying periplasmic constituents. Cytoplasmic filaments converged from their attachment points opposite the basal bodies to form arrays that ran roughly parallel to the flagellar filaments along the inner surface of the cytoplasmic membrane (CM). Motile treponemes stably attached to rabbit epithelial cells predominantly via their tips. CET revealed that T. pallidum cell ends have a complex morphology and assume at least four distinct morphotypes. Images of dividing treponemes and organisms shedding cell envelope-derived blebs provided evidence for the spirochete’s complex membrane biology. In the regions without flagellar filaments, peptidoglycan (PG) was visualized as a thin layer that divided the periplasmic space into zones of higher and lower electron densities adjacent to the CM and OM, respectively. Flagellar filaments were observed overlying the PG layer, while image modeling placed the PG-basal body contact site in the vicinity of the stator–P-collar junction. Bioinformatics and homology modeling indicated that the MotB proteins of T. pallidum, Treponema denticola, and Borrelia burgdorferi have membrane topologies and PG binding sites highly similar to those of their well-characterized Escherichia coli and Helicobacter pylori orthologs. Collectively, our results help to clarify fundamental differences in cell envelope ultrastructure between spirochetes and gram-negative bacteria. They also confirm that PG stabilizes the flagellar motor and enable us to propose that in most spirochetes motility results from rotation of the flagellar filaments against the PG