Recruitment of the TolA Protein to Cell Constriction Sites in Escherichia coli via Three Separate Mechanisms, and a Critical Role for FtsWI Activity in Recruitment of both TolA and TolQ.

The Tol-Pal system of Gram-negative bacteria helps maintain the integrity of the cell envelope and ensures that invagination of the envelope layers during cell fission occurs in a well-coordinated manner. In Escherichia coli, the five Tol-Pal proteins (TolQ, -R, -A, and -B and Pal) accumulate at cell constriction sites in a manner that normally requires the activity of the cell constriction initiation protein FtsN. While septal recruitment of TolR, TolB, and Pal also requires the presence of TolQ and/or TolA, the latter two can recognize constriction sites independently of the other system proteins.

A two-track model for the spatiotemporal coordination of bacterial septal cell wall synthesis revealed by single-molecule imaging of FtsW.

Synthesis of septal peptidoglycan (sPG) is crucial for bacterial cell division. FtsW, an indispensable component of the cell division machinery in all walled bacterial species, was recently identified in vitro as a peptidoglycan glycosyltransferase (PGTase). Despite its importance, the septal PGTase activity of FtsW has not been demonstrated in vivo.

Redesigning an International Orthopaedic CME Course: The Effects on Participant Engagement over 5 Years.

The time required to observe changes in participant evaluation of continuing medical education (CME) courses in surgical fields is unclear. We investigated the time required to observe changes in participant evaluation of an orthopaedic course after educational redesign using aggregate course-level data obtained from 1359 participants who attended one of 23 AO Davos Courses over a 5-year period between 2007 and 2011. Participants evaluated courses using two previously validated, 5-point Likert scales based on content and faculty performance, and we compared results between groups that underwent educational redesign incorporating serial needs assessment, problem-based learning, and faculty training initiatives (Masters Course), and those that did not (Non-Masters Course).

Roles of the DedD Protein in Cell Constriction.

Two key tasks of the bacterial septal-ring (SR) machinery during cell constriction are the generation of an inward-growing annulus of septal peptidoglycan (sPG) and the concomitant splitting of its outer edge into two layers of polar PG that will be inherited by the two new cell ends. FtsN is an essential SR protein that helps trigger the active constriction phase in by inducing a self-enhancing cycle of processes that includes both sPG synthesis and splitting and that we refer to as the sPG loop. DedD is an SR protein that resembles FtsN in several ways.

Roles for both FtsA and the FtsBLQ subcomplex in FtsN-stimulated cell constriction in Escherichia coli.

Escherichia coli FtsN is a bitopic membrane protein that is essential for triggering active cell constriction. A small periplasmic subdomain ((E) FtsN) is required and sufficient for function, but its mechanism of action is unclear. We isolated extragenic (E) FtsN*-suppressing mutations that restore division in cells producing otherwise non-functional variants of FtsN.

Analysis of MreB interactors in Chlamydia reveals a RodZ homolog but fails to detect an interaction with MraY.

Chlamydia is an obligate intracellular bacterial pathogen that has significantly reduced its genome in adapting to the intracellular environment. One class of genes for which the bacterium has few annotated examples is cell division, and Chlamydia lacks FtsZ, a central coordinator of the division apparatus. We have previously implicated MreB as a potential substitute for FtsZ in Chlamydia (Ouellette et al.

Following drug uptake and reactions inside Escherichia coli cells by Raman microspectroscopy.

Raman microspectroscopy combined with Raman difference spectroscopy reveals the details of chemical reactions within bacterial cells. The method provides direct quantitative data on penetration of druglike molecules into Escherichia coli cells in situ along with the details of drug-target reactions. With this label-free technique, clavulanic acid and tazobactam can be observed as they penetrate into E.

Measurement and 3D-visualization of cell-cycle length using double labelling with two thymidine analogues applied in early heart development.

Organ development is a complex spatial process in which local differences in cell proliferation rate play a key role. Understanding this role requires the measurement of the length of the cell cycle at every position of the three-dimensional (3D) structure. This measurement can be accomplished by exposing the developing embryo to two different thymidine analogues for two different durations immediately followed by tissue fixation.

Angulated locking plate in periprosthetic proximal femur fractures: biomechanical testing of a new prototype plate.

Introduction: To improve proximal plate fixation of periprosthetic femur fractures, a prototype locking plate with proximal posterior angulated screw positioning was developed and biomechanically tested.

Methods: Twelve fresh frozen, bone mineral density matched human femora, instrumented with cemented hip endoprosthesis were osteotomized simulating a Vancouver B1 fracture. Specimens were fixed proximally with monocortical (LCP) or angulated bicortical (A-LCP) head-locking screws.

Growth of the developing mouse heart: an interactive qualitative and quantitative 3D atlas.

Analysis of experiments aimed at understanding the genetic mechanisms of differentiation and growth of the heart, calls for detailed insights into cardiac growth and proliferation rate of myocytes and their precursors. Such insights in mouse heart development are currently lacking. We quantitatively assessed the 3D patterns of proliferation in the forming mouse heart and in the adjacent splanchnic mesoderm, from the onset of heart formation till the developed heart at late gestation.

Direct membrane binding by bacterial actin MreB.

Bacterial actin MreB is one of the key components of the bacterial cytoskeleton. It assembles into short filaments that lie just underneath the membrane and organize the cell wall synthesis machinery. Here we show that MreB from both T.

Identification of Escherichia coli ZapC (YcbW) as a component of the division apparatus that binds and bundles FtsZ polymers.

Assembly of the cell division apparatus in bacteria starts with formation of the Z ring on the cytoplasmic face of the membrane. This process involves the accumulation of FtsZ polymers at midcell and their interaction with several FtsZ-binding proteins that collectively organize the polymers into a membrane-associated ring-like configuration. Three such proteins, FtsA, ZipA, and ZapA, have previously been identified in Escherichia coli.

Advances in understanding E. coli cell fission.

Much of what we know about cytokinesis in bacteria has come from studies with Escherichia coli, and efforts to comprehensively understand this fundamental process in this organism continue to intensify. Major recent advances include in vitro assembly of a membrane-tethered version of FtsZ into contractile rings in lipid tubules, in vitro dynamic patterning of the Min proteins and a deeper understanding of how they direct assembly of the FtsZ-ring to midcell, the elucidation of structures, biochemical activities and interactions of other key components of the cell fission machinery, and the uncovering of additional components of this machinery with often redundant but important roles in invagination of the three cell envelope layers.

Bacterial actin MreB assembles in complex with cell shape protein RodZ.

Bacterial actin homologue MreB is required for cell shape maintenance in most non-spherical bacteria, where it assembles into helical structures just underneath the cytoplasmic membrane. Proper assembly of the actin cytoskeleton requires RodZ, a conserved, bitopic membrane protein that colocalises to MreB and is essential for cell shape determination. Here, we present the first crystal structure of bacterial actin engaged with a natural partner and provide a clear functional significance of the interaction.

Self-enhanced accumulation of FtsN at Division Sites and Roles for Other Proteins with a SPOR domain (DamX, DedD, and RlpA) in Escherichia coli cell constriction.

Of the known essential division proteins in Escherichia coli, FtsN is the last to join the septal ring organelle. FtsN is a bitopic membrane protein with a small cytoplasmic portion and a large periplasmic one. The latter is thought to form an alpha-helical juxtamembrane region, an unstructured linker, and a C-terminal, globular, murein-binding SPOR domain.

Functional outcomes after nonoperative management of fractures of the proximal humerus.

Background: Prospective follow-up data after nonoperative treatment for fractures of the proximal humerus are scarce. We studied functional outcomes and rates of complication and failure after conservative management of these common injuries.

Materials And Methods: Consecutive patients aged older than 18 years presenting to the emergency department of a large district hospital with an isolated, closed proximal humeral fracture considered suitable for functional treatment by the surgeon on charge were enrolled in a prospective, externally monitored observational study.

RodZ (YfgA) is required for proper assembly of the MreB actin cytoskeleton and cell shape in E. coli.

The bacterial MreB actin cytoskeleton is required for cell shape maintenance in most non-spherical organisms. In rod-shaped cells such as Escherichia coli, it typically assembles along the long axis in a spiral-like configuration just underneath the cytoplasmic membrane. How this configuration is controlled and how it helps dictate cell shape is unclear.

A caudal proliferating growth center contributes to both poles of the forming heart tube.

Recent studies have shown that the primary heart tube continues to grow by addition of cells from the coelomic wall. This growth occurs concomitantly with embryonic folding and formation of the coelomic cavity, making early heart formation morphologically complex. A scarcity of data on localized growth parameters further hampers the understanding of cardiac growth.

Conditional lethality, division defects, membrane involution, and endocytosis in mre and mrd shape mutants of Escherichia coli.

Maintenance of rod shape in Escherichia coli requires the shape proteins MreB, MreC, MreD, MrdA (PBP2), and MrdB (RodA). How loss of the Mre proteins affects E. coli viability has been unclear.