Isolation and characterization of the outer membrane of Escherichia coli with autodisplayed Z-domains

Abstract“Autodisplay technology” is an expression technique used to display the various recombinant proteins on the outer membrane (OM) of Escherichia coli. The resulting autodisplayed Z-domain has been used to improve the sensitivity of immunoassays. In this work, a facile isolation method of the OM fraction of E. coli with autodisplayed Z-domains was presented using (1) an enzyme reaction for the hydrolysis of the peptidoglycan layer and (2) short centrifugation steps. The purity of the isolated OM fraction was analyzed. For the estimation of contamination with bacterial proteins from other parts of E. coli, Western blots of marker proteins for the OM (OmpA), periplasm (β-lactamase), inner membrane (SecA), and cytoplasm (β-galactosidase) were performed. Additionally, assays of marker components or enzymes from each part of E. coli were carried out including the OM (KDO), inner membrane (NADH oxidase), periplasm (β-lactamase), and cytoplasm (β-galactosidase). The yield of OM isolation using this new method was determined to be 80% of the total OM amount, with less than 1% being contaminants from other parts of E. coli

2,9,16,19,22,25-Hexaoxatetra­cyclo­[24.4.0.24,7.010,15]dotriaconta-1(26),4,6,10(15),11,13,27,29,31-nona­ene

The title 22-crown-6 unit, C26H28O6, comprising of three benzo groups and triethyl­ene glycol, was prepared by the reaction of α,α′-dibromo-p-xylene with 1,8-bis­(2-hydroxy­phen­oxy)-3,6-dioxaoctane in the presence of Cs2CO3 with tetra­hydro­furan (THF) and recrystallized from dichloro­methane–hexane (1:20 v/v) at room temperature. In the mol­ecular structure, two O atoms of the central ethyl­ene glycol in the triethyl­ene glycol unit exhibit exo conformations due to intra­molecular C—H⋯O inter­actions. A number of C—H⋯O and C—H⋯π inter­molecular inter­actions contribute to the stabilization of the crystal packing

Trends and Associated Factors of Use of Opioid, Heroin, and Cannabis Among Patients for Emergency Department Visits in Nevada: 2009–2017

To examine trends and contributing factors of opioid, heroin, and cannabis-associated emergency department (ED) visits in Nevada. The 2009 to 2017 Nevada State ED database (n = 7,950,554 ED visits) were used. Use of opioid, heroin, and cannabis, respectively, was identified by the International Classification of Diseases, 9th & 10th Revisions. Three multivariable models, one for each of the 3 dependent variables, were conducted. Independent variables included year, insurance status, race/ethnicity, use of other substance, and mental health conditions. The number of individuals with opioid, heroin, cannabis-associated ED visits increased 3%, 10%, and 23% annually from 2009 to 2015, particularly among 21 to 29 age group, females, and African Americans. Use of other substance (odds ratio [OR] = 3.91; 95% confidence interval [CI] = 3.84, 3.99; reference - no use of other substance), mental health conditions (OR = 2.48; 95% CI = 2.43, 2.53; reference – without mental health conditions), Medicaid (OR = 1.41; 95% CI = 1.38, 1.44; reference – non-Medicaid), Medicare (OR = 1.44; 95% CI = 1.39, 1.49; reference – non-Medicare) and uninsured patients (OR = 1.52; 95% CI = 1.49, 1.56; reference - insured) were predictors of all three substance-associated ED visits. With a steady increase in trends of opioid, heroin, and cannabis-associated ED visits in recent years, the main contributing factors include patient sociodemographic factors, mental health conditions, and use of other substances

Surface currents from hourly variations of suspended particulate matter from Geostationary Ocean Color Imager data

Surface currents in Korean coastal regions were obtained using the maximum cross-correlation method applied to hourly suspended particulate matter images from the Geostationary Ocean Color Imager. Preliminary current vectors were filtered out by applying a series of quality-control procedures. The current vectors resulting from the tests were compared with the currents from a numerical model with tide and wind field. It was found that the estimated currents were more similarly to the currents caused by both tide and wind. A high degree of discrepancy was detected in regions of strong tidal currents, where the fundamental assumption of horizontal movement was limited due to the dominant vertical tidal mixing in the shallow region. The hourly rotations of the current vectors within a day were clarified by a comparison of the time-varying orientation angles of tidal ellipses. This study emphasized how to understand the short-term surface flows from hourly high-resolution geostationary satellite images

Flightless-I Controls Fat Storage in Drosophila

Triglyceride homeostasis is a key process of normal development and is essential for the maintenance of energy metabolism. Dysregulation of this process leads to metabolic disorders such as obesity and hyperlipidemia. Here, we report a novel function of the Drosophila flightless-I (fliI) gene in lipid metabolism. Drosophila fliI mutants were resistant to starvation and showed increased levels of triglycerides in the fat body and intestine, whereas fliI overexpression decreased triglyceride levels. These flies suffered from metabolic stress indicated by increased levels of trehalose in hemolymph and enhanced phosphorylation of eukaryotic initiation factor 2 alpha (eIF2??). Moreover, upregulation of triglycerides via a knockdown of fliI was reversed by a knockdown of desat1 in the fat body of flies. These results indicate that fliI suppresses the expression of desat1, thereby inhibiting the development of obesity; fliI may, thus, serve as a novel therapeutic target in obesity and metabolic diseases

The impact of non-pharmaceutical interventions on premature births during the COVID-19 pandemic: a nationwide observational study in Korea

BackgroundNon-pharmaceutical interventions (NPIs), such as social distancing and hand washing, have been associated with a decline in the preterm birth rate worldwide. We aimed to evaluate whether the preterm birth rate in Korea during the coronavirus disease 2019 lockdown has changed compared to that in previous years.MethodA birth registry from the Korea Statistical Information Service, which is a nationwide official database, was used to include all births claimed to have occurred between 2011 and 2020. Newborns with gestational age (GA) less than 22 weeks and birth weight less than 220 g were excluded. The pre-NPI period was designated as January 2011 to January 2020, and the NPI period was defined as February 2020 to December 2020. We assessed the effect of NPI on the incidence of prematurity per 100 births using an interrupted time-series quasi-experimental design and implementing an autoregressive integrated moving average (ARIMA) model.ResultsFrom 2011 to 2020, a total of 3,931,974 live births were registered, among which 11,416 were excluded. Consequently, the final study population included 3,920,558 live births (both singleton and multiple births) among which 275,009 (7.0%) were preterm. The preterm birth rate was significantly higher during the NPI period (8.68%) compared to that in the pre-NPI period (6.92%) (P < 0.001). The ARIMA model showed that in all singleton and multiple births, except those in July (observed 9.24, expected 8.54, [95% prediction interval {PI} 8.13–8.96], percent difference 7.81%), September (observed 7.89, expected 8.35, [95% PI 7.93–8.76], percent difference −5.66%), and December (observed 9.90, expected 9.40, [95% PI 8.98–9.82], percent difference 5.2%), most observed values were within the 95% PI of the expected values and showed an increasing trend.ConclusionIn this nationwide observational study, the trend in premature birth rate did not significantly change due to NPI implementation in Korea, as it had been increasing since 2011. The trend of Korea's birth rate appears to be unaffected by the implementation of NPIs; however, further studies with a longer follow-up period are needed

Controller optimization with constraints on probabilistic peak responses

Peak response is a more suitable index than mean response in the light of structural safety. In this study, a controller optimization method is proposed to restrict peak responses of building structures subject to earthquake excitations, which are modeled as partially stationary stochastic process. The constraints are given with specified failure probabilities of peak responses. LQR is chosen to assure stability in numerical process of optimization. Optimization problem is formulated with weightings on controlled outputs as design variables and gradients of objective and constraint functions are derived. Full state feedback controllers designed by the proposed method satisfy various design objectives and output feedback controllers using LQG also yield similar results without significant performance deterioration.The work presented in this paper was partially supported by Research Fund of the National Research Laboratory Program (Project No. M1-0203-00-0068) from the Ministry of Science and Technology in Korea. The authors also gratefully acknowledge the support of this research by the Smart Infra-Structure Technology Center (SISTeC) (Project No. R11-2022-101-03004-0(2002))