A novel bleb-dependent polysaccharide export system in nitrogen-fixing Azotobacter vinelandii subjected to low nitrogen gas levels

The alginate biofilm-producing bacterium Azotobacter vinelandii aerobically fixes nitrogen by oxygensensitivenitrogenases. Here we investigated the bacterial response to nitrogen/oxygen gas mixtures. A. vinelandii cellswere cultured in nitrogen-free minimal media containing gas mixtures differing in their ratios of nitrogen and oxygen.The bacteria did not grow at oxygen concentrations >75 % but grew well in the presence of 5 % nitrogen/25 % oxygen.Growth of wild-type and alginate-deficient strains when cultured with 50 % oxygen did not differ substantially, indicatingthat alginate is not required for the protection of nitrogenases from oxygen damage. In response to decreasing nitrogenlevels, A. vinelandii produced greater amounts of alginate, accompanied by the formation of blebs on the cell surface. Theencystment of vegetative cells occurred in tandem with the release of blebs and the development of a multilayered exine.Immunoelectron microscopy using anti alginate-antibody revealed that the blebs contained alginate molecules. Bycontrast, alginate-deficient mutants could not form blebs. Taken together, our data provide evidence for a novel blebdependentpolysaccharide export system in A. vinelandii that is activated in response to low nitrogen gas levels. [IntMicrobiol 2013; 16(1):35-44

Spatiotemporal expression of TRPM4 in the mouse cochlea

The present study was conducted to elucidate the presence of the transient receptor potential cation channel subfamily M member 4, TRPM4, in the mouse inner ear. TRPM4 immunoreactivity (IR) was found in the cell body of inner hair cells (IHCs) in the organ of Corti in the apical side of marginal cells of the stria vascularis, in the apical portion of the dark cells of the vestibule, and in a subset of the type II neurons in the spiral ganglion. Subsequently, changes in the distribution and expression of TRPM4 in the inner ear during embryonic and postnatal developments were also evaluated. Immunohistochemical localization demonstrated that the emergence of the TRPM4-IR in IHCs occurs shortly before the onset of hearing, whereas that in the marginal cells happens earlier, at the time of birth, coinciding with the onset of endolymph formation. Furthermore, semiquantitative real-time PCR assay showed that expressions of TRPM4 in the organ of Corti and in the stria vascularis increased dramatically at the onset of hearing. Because TRPM4 is a Ca2+-activated monovalent-selective cation channel, these findings imply that TRPM4 contributes to potassium ion transport, essential for the signal transduction in IHCs and the formation of endolymph by marginal cells. © 2014 Wiley Periodicals, Inc

Essential role of autoactivation circuitry on Aurora B-mediated H2AX-pS121 in mitosis

Shimada, M., Goshima, T., Matsuo, H. et al. Essential role of autoactivation circuitry on Aurora B-mediated H2AX-pS121 in mitosis. Nat Commun 7, 12059 (2016). https://doi.org/10.1038/ncomms1205

A Method to Measure Visual Field Sensitivity at the Edges of Glaucomatous Scotomata

METHODS. Subjects comprised 22 glaucomatous patients. Gradients of sensitivity were calculated for ''squares'' of test points in a patient's 24-2/30-2 VF results, so that the edges of scotomata could be identified where gradients were steep. Next, 10 new VF points were placed in these locations for each patient. Each patient's VF was then measured using this novel test grid (52 standard 24-2 test points and 10 additional points examined concurrently) on two separate occasions. The absolute difference between the measured sensitivity at each new additional test point and the average of the sensitivities of its surrounding four test points was calculated (D ave ). The intra-and intervisit reproducibility of the additional test points' thresholds was calculated. Finally, fluctuation of overall VF damage was estimated using the intraclass correlation coefficient (ICC) and the coefficient of variation (CV). RESULTS. The average of the sensitivities (D ave ) increased as the gradient of the plane steepened, whereas the reproducibility of the additional test points' thresholds remained stable. ICC was significantly higher and CV was significantly lower for the novel test grid compared with the standard 24-2 test pattern. CONCLUSIONS. It may be advantageous to increase the density of VF test points where there are large local differences in VF sensitivity. These additional measurements may result in more reproducible and well-defined estimates of scotomata

New diagnostic criteria and severity assessment of acute cholangitis in revised Tokyo guidelines

Background: The Tokyo Guidelines for the management of acute cholangitis and cholecystitis were published in 2007 (TG07) and have been widely cited in the world literature. Because of new information that has been published since 2007, we organized the Tokyo Guidelines Revision Committee to conduct a multicenter analysis to develop the updated Tokyo Guidelines (TG13). Methods/materials : We retrospectively analyzed 1,432 biliary disease cases where acute cholangitis was suspected. The cases were collected from multiple tertiary care centers in Japan. The 'gold standard' for acute cholangitis in this study was that one of the three following conditions was present: (1) purulent bile was observed; (2) clinical remission following bile duct drainage; or (3) remission was achieved by antibacterial therapy alone, in patients in whom the only site of infection was the biliary tree. Comparisons were made for the validity of each diagnostic criterion among TG13, TG07 and Charcot's triad. Results: The major changes in diagnostic criteria of TG07 were re-arrangement of the diagnostic items and exclusion of abdominal pain from the diagnostic list. The sensitivity improved from 82.8 % (TG07) to 91.8 % (TG13). While the specificity was similar to TG07, the false positive rate in cases of acute cholecystitis was reduced from 15.5 to 5.9 %. The sensitivity of Charcot's triad was only 26.4 % but the specificity was 95.6 %. However, the false positive rate in cases of acute cholecystitis was 11.9 % and not negligible. As for severity grading, Grade II (moderate) acute cholangitis is defined as being associated with any two of the significant prognostic factors which were derived from evidence presented recently in the literature. The factors chosen allow severity assessment to be performed soon after diagnosis of acute cholangitis. Conclusion: TG13 present a new standard for the diagnosis, severity grading, and management of acute cholangitis. © 2012 The Author(s).link_to_subscribed_fulltex

New diagnostic criteria and severity assessment of acute cholecystitis in revised Tokyo guidelines

Background: The Tokyo Guidelines for the management of acute cholangitis and cholecystitis (TG07) were published in 2007 as the world's first guidelines for acute cholangitis and cholecystitis. The diagnostic criteria and severity assessment of acute cholecystitis have since been widely used all over the world. A validation study of TG07 has shown that the diagnostic criteria for acute cholecystitis are highly reliable but that the definition of definite diagnosis is ambiguous. In addition, considerable new evidence referring to acute cholecystitis as well as evaluations of TG07 have been published. Consequently, we organized the Tokyo Guidelines Revision Committee to evaluate TG07, recognize new evidence, and conduct a multi-center analysis to revise the guidelines (TG13). Methods and materials: We retrospectively analyzed 451 patients with acute cholecystitis from multiple tertiary care centers in Japan. All 451 patients were first evaluated using the criteria in TG07. The "gold standard" for acute cholecystitis in this study was a diagnosis by pathology. The validity of TG07 diagnostic criteria was investigated by comparing clinical with pathological diagnosis. Results: Of 451 patients evaluated, a total of 227 patients were given a diagnosis of acute cholecystitis by pathological examination (prevalence 50.3 %). TG07 criteria provided a definite diagnosis of acute cholecystitis in 224 patients. The sensitivity of TG07 diagnostic criteria for acute cholecystitis was 92.1 %, and the specificity was 93.3 %. Based on the preliminary results, new diagnostic criteria for acute cholecystitis were proposed. Using the new criteria, the sensitivity of definite diagnosis was 91.2 %, and the specificity was 96.9 %. The accuracy rate was improved from 92.7 to 94.0 %. In regard to severity grading among 227 patients, 111 patients were classified as Mild (Grade I), 104 as Moderate (Grade II), and 12 as Severe (Grade III). Conclusion: The proposed new diagnostic criteria achieved better performance than the diagnostic criteria in TG07. Therefore, the proposed criteria have been adopted as new diagnostic criteria for acute cholecystitis and are referred to as the 2013 Tokyo Guidelines (TG13). Regarding severity assessment, no new evidence was found to suggest that the criteria in TG07 needed major adjustment. As a result, TG07 severity assessment criteria have been adopted in TG13 with minor changes. © 2012 The Author(s).link_to_subscribed_fulltex