Ottesensantana0764

(-)-5-Epieremophilene, an epimer of the versatile sesquiterpene (+)-valencene, is an inaccessible natural product catalyzed by three sesquiterpene synthases (SmSTPSs1-3) of the Chinese medicinal herb Salvia miltiorrhiz , and its biological activity remains less explored. In this study, three metabolically engineered Escherichia coli strains were constructed for (-)-5-epieremophilene production, with yields of 42.4-76.0 mg/L in shake-flask culture. Introducing an additional copy of farnesyl diphosphate synthase (FDPS) gene through fusion expression of SmSTPS1-FDPS or dividing the FDP synthetic pathway into two modules resulted in significantly improved the production, and ultimately 250 mg of (-)-5-epieremophilene was achieved. Biological assay indicated that (-)-5-epieremophilene showed significant antifeedant activity against Helicoverpa armigera (EC 50 = 1.25 μg/cm 2 ), a common pest of S. miltiorrhiza , implying its potential defensive role in the plant. The results provided an ideal material supply for studying other potential biological activities of (-)-5-epieremophilene, and also a strategy for manipulating terpene production in engineered E. coli using synthetic biology. © 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.Human Guanylate-Binding Protein 1 (hGBP-1) shows a dimer-induced acceleration of the GTPase activity yielding GDP as well as GMP. While the head-to-head dimerization of the large GTPase (LG) domain is well understood, the role of the rest of the protein, particularly of the GTPase effector domain (GED), in dimerization and GTP hydrolysis is still obscure. In this study, with truncations and point mutations on hGBP-1 and by means of biochemical and biophysical methods, we demonstrate that the intramolecular communication between the LG domain and the GED (LGGED) is crucial for protein dimerization and dimer-stimulated GTP hydrolysis. In the course of GTP binding and γ-phosphate cleavage, conformational changes within hGBP-1 are controlled by a chain of amino acids ranging from the region near the nucleotide binding pocket to the distant LGGED interface and lead to the release of the GED from the LG domain. click here This opening of the structure allows the protein to form GEDGED contacts within the dimer, in addition to the established LGLG interface. After releasing the cleaved γ-phosphate, the dimer either dissociates yielding GDP as the final product or it stays dimeric to further cleave the β-phosphate yielding GMP. The second phosphate cleavage step, i.e. the formation of GMP, is even more strongly coupled to structural changes and thus more sensitive to structural restraints imposed by the GED. Altogether, we depict a comprehensive mechanism of GTP hydrolysis catalyzed by hGBP-1, which provides a detailed molecular understanding of the enzymatic activity connected to large structural rearrangements of the protein. This article is protected by copyright. All rights reserved.BACKGROUND AND AIMS Mixed hepatocellular-cholangiocarcinoma (HCC-CC) is a biphenotypic liver cancer thought to have unfavorable tumor biology and a poor prognosis. Surgical outcomes of HCC-CC remain unclear. We aimed to evaluate the clinical and characteristics and surgical outcomes of HCC-CC. APPROACH AND RESULTS Case series of patients undergoing resection for HCC-CC (n=47), hepatocellular carcinoma (HCC) (n=468), and intrahepatic cholangiocarcinoma (ICC) (N=108) at a single Western center between 2001 and 2015. Cases of HCC-CC were matched to cases of HCC and ICC on important clinical factors including tumor characteristics (size, vascular invasion, differentiation), and underlying cirrhosis. Patients with HCC-CC had rates of viral hepatitis comparable to patients with HCC (78.5% vs. 80%) and 42.5% had underlying cirrhosis. When matched on tumor size, HCC-CC was more poorly differentiated than HCC (68.3% vs. 27.3%, respectively; P less then 0.001) and ICC (68.3% vs. 34.8%, respectively, P=0.01) but had similar post-resection survival (5-year survival HCC-CC 49.7%, HCC 54.8%, ICC 68.7%, P=0.61) and recurrence (3-year recurrence HCC-CC 57.9%, HCC 61.5%, and ICC 56%, P=0.55). Outcomes were similar between HCC-CC and HCC when matched on underlying cirrhosis and tumor size. Cancer type was not predictive of survival or tumor recurrence. CONCLUSIONS Survival after resection of HCC-CC is similar to HCC when matched for tumor size, despite HCC-CC tumors being more poorly differentiated. Exclusion of HCC-CC from management strategies recommended for HCC, including consideration for liver transplantation, may not be warranted. This article is protected by copyright. All rights reserved.Although mice are widely used to elucidate factors contributing to penile disorders and develop treatment options, quantification of tissue changes upon intervention is either limited to minuscule tissue volume (histology) or acquired with limited spatial resolution (MRI/CT). Thus, imaging method suitable for expeditious acquisition of the entire mouse penis with subcellular resolution is described that relies on both aqueous- (CUBIC) and solvent-based (FDISCO) tissue optical clearing (TOC). The combined TOC approach allows to image mouse penis innervation and vasculature with unprecedented detail and, for the first time, reveals the three-dimensional structure of murine penis fibrocartilage. This article is protected by copyright. All rights reserved. This article is protected by copyright. All rights reserved.OBJECTIVE Adoption of emergency department (ED)-initiation of buprenorphine (BUP) for opioid use disorder (OUD) into routine emergency care has been slow, partly due to clinicians' unfamiliarity with this practice and perceptions that it is complicated and time-consuming. To address these barriers and guide emergency clinicians through the process of BUP initiation, we implemented a user-centered computerized clinical decision support system (CDS). This study was conducted to assess the feasibility of implementation and to evaluate the preliminary efficacy of the intervention to increase the rate of ED-initiated BUP. METHODS An interrupted time series study was conducted in an urban, academic ED from April 2018-February 2019 (pre-implementation phase), March 2019-August 2019 (implementation phase), and September 2019-December 2019 (maintenance phase) to study the effect of the intervention on adult ED patients identified by a validated EHR-based computable phenotype consisting of structured data consistent wiates, the relative risk of BUP initiation rate was 2.73 (95% CI 0.62, 12.0; p=0.18). Similarly, the number of unique attendings who initiated BUP increased modestly 7/53 (13.0%) to 13/57 (22.8%, p=0.10) after offering just-in-time training during the implementation period. The rate of naloxone prescribed at discharge also increased (6.5%, pre-implementation; 11.5%, implementation; p less then 0.01). The intervention received a System Usability Scale score of 82.0 (95% CI 76.7-87.2). CONCLUSION Implementation of user-centered CDS at a single ED was feasible, acceptable, and associated with increased rates of ED-initiated BUP and naloxone prescribing in patients with OUD and a doubling of the number of unique physicians adopting the practice. We have implemented this intervention across several health systems in an ongoing trial to assess its effectiveness, scalability, and generalizability. This article is protected by copyright. All rights reserved.A copper-catalyzed electrophilic etherification of arylboronic esters is reported. Isoxazolidines are utilized as easily available and stable [RO] + surrogates to give 1,3-amino aryl ethers. The O -selective arylation of isoxazolidines takes place without causing competitive N -arylation. In contrast to previously reported anionic conditions, our copper-catalyzed conditions are mild enough to achieve high functional group tolerance. Preliminary mechanistic studies and DFT calculations support that the reaction proceeds via a transmetalation/oxidative addition pathway, followed by a Lewis acid-promoted reductive elimination to induce the crucial O -selectivity. © 2020 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.BACKGROUND Hospital affiliated freestanding emergency departments (FREDs) are rapidly proliferating in some states and have been the subject of recent policy debate. As FREDs' role in acute care delivery is expanding in certain regions, little is known about the quality of care that they provide for their sickest patients. Our aim was to compare timeliness of emergent care at FREDs and hospital-based EDs (HEDs) for patient visits with selected high acuity and time-sensitive conditions. METHODS We performed a retrospective observational analysis of adult patient visit data from 19 FREDs and five HEDs from one health system over a one-year period. Median time-to-events and hazard ratios (HRs) with 95% confidence intervals (CIs) were calculated via cox regression. RESULTS The median time to electrocardiogram for visits with chest pain was 10 minutes at FREDs and 9 minutes at HEDs [HR 0.91 (CI 0.87 - 0.96)]. Time to cardiac catheterization lab for visits with ST-segment elevation myocardial infarction (STEMI) wasved.BACKGROUND More than 90% of gastric cancer cases are caused by Helicobacter pylori infections. To prevent gastric cancer, an H pylori test-and-treat strategy targeting young people has been implemented in various places in Japan. In this study, we evaluated the effectiveness of an H pylori test-and-treat strategy for second-year junior high school students in Takatsuki City. MATERIALS AND METHODS In 2014-2017, a urine-based H pylori test was used for initial screening. The final infection status was determined by a 13 C-urea breath test (13 C-UBT). link2 Successful H pylori eradication was confirmed by 13 C-UBT 3 months after treatment. First-line eradication therapy was changed from 10 mg of rabeprazole, 750 mg of amoxicillin, and 200 mg of clarithromycin twice daily for 7 days in 2014 to 20 mg of vonoprazan, 750 mg of amoxicillin, and 200 mg of clarithromycin twice daily for 7 days in 2015-2017. Second-line eradication therapy included 10 mg of rabeprazole, 750 mg of amoxicillin, and 250 mg of metronidazole twice daily for 7 days. link3 RESULTS In total, 8067 of 13 055 students participated this project and 206 students were diagnosed with H pylori infection. The success rate of first-line therapy was 45.9% in 2014 and 83.8% after the revised first-line therapy was administered. The final eradication rate was 98.5%. There were no severe side effects. CONCLUSION Our results support the use of the H pylori test-and-treat strategy for junior high school students as a safe approach for the prevention of gastric cancer. H pylori eradication therapy with vonoprazan could be a standard therapy in children. © 2020 John Wiley & Sons Ltd.BACKGROUND The clinical presentation of European patients with mild-to-moderate Covid-19 infection is still unknown. OBJECTIVE To study the clinical presentation of Covid-19 in Europe. METHODS Patients with positive diagnosis of Covid-19 were recruited from 18 European hospitals. Epidemiological and clinical data were obtained through a standardized questionnaire. Bayesian analysis was used for analyzing the relationship between outcomes. RESULTS 1,420 patients completed the study (962 females, 30.7% of health care workers). The mean age of patients was 39.17±12.09 years. The most common symptoms were headache (70.3%), loss of smell (70.2%), nasal obstruction (67.8%), cough (63.2%), asthenia (63.3%), myalgia (62.5%), rhinorrhea (60.1%), gustatory dysfunction (54.2%) and sore throat (52.9%). Fever was reported by on 45.4%. The mean duration of Covid-19 symptoms of mild-to-moderate cured patients was 11.5±5.7 days. The prevalence of symptoms significantly varied according to age and sex. Young patients more frequently had ear, nose, and throat complaints, whereas elderly individuals often presented fever, fatigue and loss of appetite.