Wukuhn5560

Bacteriochlorophyll a (BChl) is an essential pigment for anoxygenic photosynthesis. In late steps of the BChl biosynthesis of Rhodobacter capsulatus, the C8 vinyl group and C7=C8 double bond of 8-vinyl chlorophyllide a (8 V-Chlide) are reduced by a C8 vinyl reductase (8VR), BciA, and a nitrogenase-like enzyme, chlorophyllide a oxidoreductase (COR), respectively, to produce 3-vinyl-bacteriochlorphyllide a. Recently, we discovered 8VR activity in COR. However, the kinetic parameters of the COR 8VR activity remain unknown, while those of the COR C7=C8 reductase activity and BciA have been reported. Here, we determined the kinetic parameters of COR 8VR activity by using 8 V-Chlide. The Km value for 8 V-Chlide was 1.4 μM, which is much lower than the 6.2 μM determined for the C7=C8 reduction of Chlide. The kinetic parameters of the dual activities of COR suggest that COR catalyzes the reduction of the C8 vinyl group of 8 V-Chlide preferentially over C7=C8 reduction when both substrates are supplied during BChl biosynthesis. © 2020 Wiley-VCH Verlag GmbH & Co. Rapamycin in vitro KGaA, Weinheim.Electrodes based on organic matter operating in aqueous electrolytes enable new approaches and technologies for assembling and utilizing batteries that are difficult to achieve with traditional electrode materials. Here, we report how thiophene-based trimeric structures with naphthoquinone or hydroquinone redox-active pendent groups can be processed in solution, deposited, dried and subsequently polymerized in solid state to form conductive (redox) polymer layers without any additives. Such post-deposition polymerization offers efficient use of material, high mass loading (up to 10 mg cm-2 ) and good flexibility in the choice of substrate and coating method. By employing these materials as anode and cathode in an acidic aqueous electrolyte a rocking-chair proton battery is built. The battery shows good cycling stability (85 % after 500 cycles), withstands rapid charging, with full capacity (60 mAh g-1 ) reached within 100 seconds, allows for direct integration with photovoltaics, and retains its favorable characteristics even at -24 °C. © 2020 The Authors. Published by Wiley-VCH Verlag GmbH & Co. KGaA.AIM Bowel dysfunction following anterior resection (AR) is termed 'low anterior resection syndrome (LARS)'. It is unclear whether such dysfunction occurs following other bowel/pelvic operations as well. This study aimed to characterise and compare bowel dysfunction following AR, right hemicolectomy (RH) and radical cystectomy (RC). METHOD A prospective study of consecutive patients undergoing AR, RH, and RC (2002-2012) was performed at a tertiary referral centre in Sydney, Australia. Outcome measures included (i) patient-reported (satisfaction with bowel function, self-described bowel function, and self-reported change in bowel function); (ii) objective assessment of bowel function using validated criteria to identify symptoms and stratify patients into those with constipation and/or faecal incontinence (FI); and (iii) health-related quality of life (SF-36v2). RESULTS Of 743 eligible patients, 70% participated (AR n=338, mean age 69.6yrs [SD11.9], 59% male; RH n=150, 75.8yrs [SD10.5], 54% male; RC n=34, 71.1yrs [SD14.1], 71% male). AR patients were three times more likely to report change in bowel function post-surgery, and self-judged their bowel function as 'abnormal' more frequently (64%) than following RH (35%) and RC (35%) (P less then 0.01). AR patients were four times more likely to meet criteria for concomitant constipation and FI. Patients with concomitant constipation and FI had lower physical and mental SF-36v2 scores (P less then 0.001). CONCLUSION Bowel dysfunction occurred after RH and RC but rates were higher following AR. This suggests that LARS occurs due to a direct impact of partial/complete loss of the rectum rather than just due to loss of bowel length and/or the consequence(s) of pelvic dissection. This article is protected by copyright. All rights reserved.Two DNA aptamers that bind the heparin-binding domain (HBD) of the human vascular endothelial growth factor 165 (VEGF-165) have been previously reported. Although VEGF-165 is a homodimeric protein and the two aptamers have different sequences and secondary structures, the aptamers appear to occupy the same binding site and cannot form a 2  1 aptamer/protein complex, thus making them unsuitable for creating a higher-affinity dimeric DNA aptamer. This has motivated us to conduct a new in vitro selection experiment to search for new VEGF-165-binding DNA aptamers with different properties. We undertook a multistream selection strategy in which the concentration of VEGF-165 was varied significantly. We carried out 11 rounds of selection, and next-generation sequencing was conducted for every round in each stream. From comprehensive sequence analysis, we identified four classes of DNA aptamers, of which two were reported before, but two are new DNA aptamers. One of the new aptamers exhibits a unique property that has never been observed before it is capable of forming the 2  1 aptamer/protein complex with VEGF-165. This work has expanded the repertoire of VEGF-165-binding DNA aptamers and creates a possibility to engineer a higher affinity homodimeric aptamer for VEGF-165. © 2020 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.We describe a novel, easy and efficient combinatorial phage display peptide substrate mining method to map the substrate specificity of proteases. The peptides are displayed on the pVII capsid of the M13 bacteriophage, which renders pIII necessary for infectivity and efficient retrieval, unmodified. As capture module, the 3XFLAG was chosen due to its very high binding efficiency to anti-FLAG mAbs and its independency of any posttranslational modification. This library was tested with Factor-VII activating protease (WT-FSAP) and its single-nucleotide polymorphism variant Marburg-I (MI)-FSAP. The WT-FSAP results confirmed the previously reported Arg/Lys centered FSAP cleavage site consensus as dominant, as well as reinforcing MI-FSAP as a loss of function mutant. Surprisingly, rare substrate clones devoid of basic amino acids were also identified, and indeed one of these peptides were cleaved as a free peptide suggesting a broader range of WT-FSAP substrates than previously anticipated. © 2020 WILEY-VCH Verlag GmbH & Co.