Hosts utilize macroautophagy/autophagy to clear invading bacteria; however, bacteria have also developed a specific mechanism to survive by manipulating the host cell autophagy mechanism. One pathogen, Legionella pneumophila, can hinder host cell autophagy by using the specific effector protein RavZ that cleaves phosphatidylethanolamine-conjugated LC3 on the phagophore membrane. However, the detailed molecular mechanisms associated with the function of RavZ have hitherto remained unclear. Here, we report on the biochemical characteristics of the RavZ-LC3 interaction, the solution structure of the 1:2 complex between RavZ and LC3, and crystal structures of RavZ showing different conformations of the active site loop without LC3. Based on our biochemical, structural, and cell-based analyses of RavZ and LC3, both distant flexible N-and C-terminal regions containing LC3-interacting region (LIR) motifs are important for substrate recognition. These results suggest a novel mechanism of RavZ action on the phagophore membrane and lay the groundwork for understanding how bacterial pathogens can survive autophagy.

Effectiveness of a daytime rapid response system in hospitalized surgical ward patients Background: Clinical deteriorations during hospitalization are often preventable with a rapid response system (RRS). We aimed to investigate the effectiveness of a daytime RRS for surgical hospitalized patients. Methods: A retrospective cohort study was conducted in 20 general surgical wards at a 1,779-

Kynurenine 3-monooxygenase (KMO) inhibitors have been developed for the treatment of neurodegenerative disorders. The mechanisms of flavin reduction and hydrogen peroxide production by KMO inhibitors are unknown. Herein, we report the structure of human KMO and crystal structures of Saccharomyces cerevisiae (sc) and Pseudomonas fluorescens (pf) KMO with Ro 61-8048. Proton transfer in the hydrogen bond network triggers flavin reduction in p-hydroxybenzoate hydroxylase, but the mechanism triggering flavin reduction in KMO is different. Conformational changes via π-π interactions between the loop above the flavin and substrate or non-substrate effectors lead to disorder of the C-terminal α helix in scKMO and shifts of domain III in pfKMO, stimulating flavin reduction. Interestingly, Ro 61-8048 has two different binding modes. It acts as a competitive inhibitor in scKMO and as a non-substrate effector in pfKMO. These findings provide understanding of the catalytic cycle of KMO and insight for structure-based drug design of KMO inhibitors.

Highlights d Leucine binding induces conformational change of LARS1 for RagD binding d ATP increases leucine binding to LARS1 for enhanced RagD binding d Leucine-controlled R-lever triggers long-range conformational change of LARS1 d LARS1 cycles between ''sensing on (leucine+)'' and ''sensing off (leucineÀ)'' states.

PurposeThe purpose of this study was to examine whether periodontal pocket could be satisfactorily visualized by optical coherence tomography (OCT) and to suggest quantitative methods for measuring periodontal pocket depth.MethodsWe acquired OCT images of periodontal pockets in a porcine model and determined the actual axial resolution for measuring the exact periodontal pocket depth using a calibration method. Quantitative measurements of periodontal pockets were performed by real axial resolution and compared with the results from manual periodontal probing.ResultsThe average periodontal pocket depth measured by OCT was 3.10±0.15 mm, 4.11±0.17 mm, 5.09±0.17 mm, and 6.05±0.21 mm for each periodontal pocket model, respectively. These values were similar to those obtained by manual periodontal probing.ConclusionsOCT was able to visualize periodontal pockets and show attachment loss. By calculating the calibration factor to determine the accurate axial resolution, quantitative standards for measuring periodontal pocket depth can be established regardless of the position of periodontal pocket in the OCT image.