Flavobacterium Gliding Motility: From Protein Secretion to Cell Surface Adhesin Movements

Flavobacterium johnsoniae exhibits rapid gliding motility over surfaces. At least twenty genes are involved in this process. Seven of these, gldK, gldL, gldM, gldN, sprA, sprE, and sprT encode proteins of the type IX protein secretion system (T9SS). The T9SS is required for surface localization of the motility adhesins SprB and RemA, and for secretion of the soluble chitinase ChiA. This thesis demonstrates that the gliding motility proteins GldA, GldB, GldD, GldF, GldH, GldI and GldJ are also essential for secretion. Cells with mutations in the genes encoding any of these seven proteins had normal levels of gldK mRNA but dramatically reduced levels of GldK protein, which may explain the secretion defects of the motility mutants. GldJ is necessary for stable accumulation of GldK and each mutant lacked GldJ protein. F. johnsoniae cells that produced truncated GldJ, lacking eight to thirteen amino acids from the C-terminus, accumulated GldK but were deficient in gliding motility. SprB was secreted by these cells but was not propelled along their surfaces. This C-terminal region of GldJ is thus required for gliding motility but not for secretion. The identification of mutants that are defective for motility but competent for secretion begins to untangle the F. johnsoniae gliding motility machinery from the T9SS.Proteins secreted by the T9SS typically have conserved C-terminal domains (CTDs) belonging to the type A CTD or type B CTD families. Attachment of 70 to 100 amino acid type A CTDs to a foreign protein allow its secretion. Type B CTDs are common but have received little attention. Secretion of the foreign protein sfGFP fused to regions spanning the SprB type B CTD (sfGFP-CTDSprB) was analyzed. CTDs of 218 amino acids or longer resulted in secretion of sfGFP whereas a 149 amino acid region did not. Some sfGFP was secreted in soluble form whereas the rest was attached on the cell surface. Surface-attached sfGFP was rapidly propelled along the cell, suggesting productive interaction with the motility machinery. This did not result in rapid cell movement, which apparently requires additional regions of SprB. Secretion of sfGFP-CTDSprB required coexpression with sprF, which lies downstream of sprB. SprF is similar in sequence to Porphyromonas gingivalis PorP. Most F. johnsoniae genes encoding proteins with type B CTDs lie immediately upstream of porP/sprF-like genes. sfGFP was fused to the type B CTD from one such protein (Fjoh_3952). This resulted in secretion of sfGFP only when it was coexpressed with its cognate PorP/SprF-like protein. These results highlight the need for extended regions of type B CTDs and for coexpression with the appropriate PorP/SprF-like protein for efficient secretion and cell-surface localization of cargo proteins

Alien Registration- Johnston, Joseph (Saint John Plantation, Aroostook County)

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Comments: A Poisoned Arrow in His Quiver: Why Forbidding an Entire Branch of Government from Communicating with a Reporter Violates the First Amendment

[There exists] a profound national commitment to the principle that debate on public issues should be uninhibited, robust, and wide-open, and that it may well include vehement, caustic, and sometimes unpleasantly sharp attacks on government and public officials. Nothing can now be believed which is seen in a newspaper. Truth itself becomes suspicious by being put into that polluted vehicle

Safety Videos for Major Shop Equipment in Cal Poly BRAE Lab 1

At California Polytechnic State University, San Luis Obispo, students from all majors can take a variety of classes in the BioResource and Agriculture Engineering Department (BRAE). It is essential for future high school mechanics educators to learn the basic shop tools and the importance of safety in the laboratory. In shop 1 of the BRAE Department, there are many different tools students will find in a mechanics classroom. The five main power tools are the Table Saw, Drill Press, Bench Grinder, Radial Arm Saw, and Disk/Belt Sander. In BRAE 121 Agricultural Mechanics, students learn basic shop skills about lab safety and proper tool usage. From time to time the instructor gives directions on how to use particular tools in order to be safe. But, with busy schedules and unexpected events, students may miss a tutorial on how to use a particular machine. Students need alternative resources to easily get caught up on how to safely use shop equipment and this project will create a safety video for future BRAE Lab 1 use

Exploring and improving the escalation of care process for deteriorating patients on surgical wards in UK hospitals

Despite impressive progress in technical skills, the rate of adverse events in surgery remains unfavourably high. The variation seen in surgical outcomes may be dependent on the quality of ward-based surgical care provided to post-operative patients with complications, specifically, the recognition, communication and response to patient deterioration. This process can be termed escalation of care and is an under-explored area of surgical research. This thesis demonstrates the impact of delays in the escalation of care process on patient outcome. The facilitators of, and barriers to, escalation of care are then identified and described in the context of the UK surgical department. In order to prioritise areas within the escalation of care process amenable to intervention, a systematic risk assessment was conducted revealing suboptimal communication technology and a lack of human factors education as key failures. To ensure that communication technology intervention was conducted based on evidence, several exploratory studies describe the current methods of communication in surgery and explore areas of innovation and intervention. Following this, a human factors intervention bundle was implemented within a busy surgical department, which successfully improved supervision, escalation of care and safety culture. This thesis describes, for the first time, escalation of care in surgery and outlines important strategies for intervention in this safety-critical process. To date, ward-based care has been one of the most under-researched areas in surgery, despite its clear importance. The tools to improve escalation of care in surgery have been described and initial attempts at implementation have demonstrated great promise. Future use of these strategies should benefit surgeons and other clinical staff of all grades and ultimately, the surgical patient.Open Acces

Comments: A Poisoned Arrow in His Quiver: Why Forbidding an Entire Branch of Government from Communicating with a Reporter Violates the First Amendment

[There exists] a profound national commitment to the principle that debate on public issues should be uninhibited, robust, and wide-open, and that it may well include vehement, caustic, and sometimes unpleasantly sharp attacks on government and public officials. Nothing can now be believed which is seen in a newspaper. Truth itself becomes suspicious by being put into that polluted vehicle

Accurals quality and price synchronicity

This study examines the relation between accruals quality and price synchronicity, a measure of the relative amount of firm-specific information reflected in price. Higher accruals quality imply better quality earnings news, hence, more firm-specific information is incorporated into price for firms with higher accruals quality. More firm-specific information reduces price synchronicity, hence, we hypothesize a negative relation between accruals quality and price synchronicity. On the other hand, literature shows that accruals quality reduces idiosyncratic volatility which tends to be negatively correlated with price synchronicity. If the latter effects dominate the relation between accruals quality and price synchronicity, we should observe a positive relationship between accruals quality and price synchronicity. Controlling for idiosyncratic volatility, we find a significant negative relation between accruals quality and price synchronicity after controlling for idiosyncratic volatility. We investigate this further by partitioning the sample by analyst following. If earnings information complements analysts’ information, we expect to find a stronger negative relation between accruals quality and price synchronicity for firms that are followed by analysts. If, on the other hand, earnings information and analysts’ information are substitutes, we expect to find a stronger negative relation between accruals quality and price synchronicity for those firms that are not followed by analysts. We find that accruals quality has a greater impact on price synchronicity for firms with an analyst following compared to firm that do not have an analyst following. This is consistent with the notion that earnings information complements analyst information

Regulation of human lung fibroblast alpha 1(I) procollagen gene expression by tumor necrosis factor alpha, interleukin-1 beta, and prostaglandin E2.

We investigated the participation of prostaglandin (PG) E2 in the regulation of the alpha 1(I) procollagen gene expression by tumor necrosis factor alpha (TNF alpha), and interleukin-1 beta (IL-1 beta) in normal adult human lung fibroblasts. TNF alpha (100 units/ml) and IL-1 beta (100 units/ml) stimulated the production of PGE2 and caused a dose-dependent inhibition of up to 54 and 66%, respectively, of the production of type I procollagen. Preincubation of cultures with indomethacin partially reversed the inhibition of procollagen production induced by the cytokines. Cytokine-stimulated endogenous fibroblast PG accounted for 35 and 68% of the inhibition induced by TNF alpha and IL-1 beta, respectively. Steady-state mRNA levels for alpha 1(I) procollagen paralleled the changes in collagen production. The transcription rate of the alpha 1(I) procollagen gene was reduced by 58% by TNF alpha and by 43% by IL-1 beta. Cytokine-stimulated endogenous PG production accounted for half of these effects. These results indicate that TNF alpha and IL-1 beta inhibit the expression of the alpha 1(I) procollagen gene in human lung fibroblasts at the transcriptional level by a PGE2-independent effect as well as through the effect of endogenous fibroblast PGE2 released under the stimulus of the cytokines