A Method for Rapid Screening of Anilide-Containing AMPK Modulators Based on Computational Docking and Biological Validation

Adenosine 5′-monophsphate-activated protein kinase (AMPK) is a crucial energy sensor for maintaining cellular homeostasis. Targeting AMPK may provide an alternative approach in treatment of various diseases like cancer, diabetes, and neurodegenerations. Accordingly, novel AMPK activators are frequently identified from natural products in recent years. However, most of such AMPK activators are interacting with AMPK in an indirect manner, which may cause off-target effects. Therefore, the search of novel direct AMPK modulators is inevitable and effective screening methods are needed. In this report, a rapid and straightforward method combining the use of in silico and in vitro techniques was established for selecting and categorizing huge amount of compounds from chemical library for targeting AMPK modulators. A new class of direct AMPK modulator have been discovered which are anilides or anilide-like compounds. In total 1,360,000 compounds were virtually screened and 17 compounds were selected after biological assays. Lipinski’s rule of five assessment suggested that, 13 out of the 17 compounds are demonstrating optimal bioavailability. Proton acceptors constituting the structure of these compounds and hydrogen bonds with AMPK in the binding site appeared to be the important factors determining the efficacy of these compounds

Antimicrobial resistance among migrants in Europe: a systematic review and meta-analysis

BACKGROUND: Rates of antimicrobial resistance (AMR) are rising globally and there is concern that increased migration is contributing to the burden of antibiotic resistance in Europe. However, the effect of migration on the burden of AMR in Europe has not yet been comprehensively examined. Therefore, we did a systematic review and meta-analysis to identify and synthesise data for AMR carriage or infection in migrants to Europe to examine differences in patterns of AMR across migrant groups and in different settings. METHODS: For this systematic review and meta-analysis, we searched MEDLINE, Embase, PubMed, and Scopus with no language restrictions from Jan 1, 2000, to Jan 18, 2017, for primary data from observational studies reporting antibacterial resistance in common bacterial pathogens among migrants to 21 European Union-15 and European Economic Area countries. To be eligible for inclusion, studies had to report data on carriage or infection with laboratory-confirmed antibiotic-resistant organisms in migrant populations. We extracted data from eligible studies and assessed quality using piloted, standardised forms. We did not examine drug resistance in tuberculosis and excluded articles solely reporting on this parameter. We also excluded articles in which migrant status was determined by ethnicity, country of birth of participants' parents, or was not defined, and articles in which data were not disaggregated by migrant status. Outcomes were carriage of or infection with antibiotic-resistant organisms. We used random-effects models to calculate the pooled prevalence of each outcome. The study protocol is registered with PROSPERO, number CRD42016043681. FINDINGS: We identified 2274 articles, of which 23 observational studies reporting on antibiotic resistance in 2319 migrants were included. The pooled prevalence of any AMR carriage or AMR infection in migrants was 25·4% (95% CI 19·1-31·8; I2 =98%), including meticillin-resistant Staphylococcus aureus (7·8%, 4·8-10·7; I2 =92%) and antibiotic-resistant Gram-negative bacteria (27·2%, 17·6-36·8; I2 =94%). The pooled prevalence of any AMR carriage or infection was higher in refugees and asylum seekers (33·0%, 18·3-47·6; I2 =98%) than in other migrant groups (6·6%, 1·8-11·3; I2 =92%). The pooled prevalence of antibiotic-resistant organisms was slightly higher in high-migrant community settings (33·1%, 11·1-55·1; I2 =96%) than in migrants in hospitals (24·3%, 16·1-32·6; I2 =98%). We did not find evidence of high rates of transmission of AMR from migrant to host populations. INTERPRETATION: Migrants are exposed to conditions favouring the emergence of drug resistance during transit and in host countries in Europe. Increased antibiotic resistance among refugees and asylum seekers and in high-migrant community settings (such as refugee camps and detention facilities) highlights the need for improved living conditions, access to health care, and initiatives to facilitate detection of and appropriate high-quality treatment for antibiotic-resistant infections during transit and in host countries. Protocols for the prevention and control of infection and for antibiotic surveillance need to be integrated in all aspects of health care, which should be accessible for all migrant groups, and should target determinants of AMR before, during, and after migration. FUNDING: UK National Institute for Health Research Imperial Biomedical Research Centre, Imperial College Healthcare Charity, the Wellcome Trust, and UK National Institute for Health Research Health Protection Research Unit in Healthcare-associated Infections and Antimictobial Resistance at Imperial College London

Safety and efficacy of the ChAdOx1 nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four randomised controlled trials in Brazil, South Africa, and the UK.

BACKGROUND: A safe and efficacious vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), if deployed with high coverage, could contribute to the control of the COVID-19 pandemic. We evaluated the safety and efficacy of the ChAdOx1 nCoV-19 vaccine in a pooled interim analysis of four trials. METHODS: This analysis includes data from four ongoing blinded, randomised, controlled trials done across the UK, Brazil, and South Africa. Participants aged 18 years and older were randomly assigned (1:1) to ChAdOx1 nCoV-19 vaccine or control (meningococcal group A, C, W, and Y conjugate vaccine or saline). Participants in the ChAdOx1 nCoV-19 group received two doses containing 5 × 1010 viral particles (standard dose; SD/SD cohort); a subset in the UK trial received a half dose as their first dose (low dose) and a standard dose as their second dose (LD/SD cohort). The primary efficacy analysis included symptomatic COVID-19 in seronegative participants with a nucleic acid amplification test-positive swab more than 14 days after a second dose of vaccine. Participants were analysed according to treatment received, with data cutoff on Nov 4, 2020. Vaccine efficacy was calculated as 1 - relative risk derived from a robust Poisson regression model adjusted for age. Studies are registered at ISRCTN89951424 and ClinicalTrials.gov, NCT04324606, NCT04400838, and NCT04444674. FINDINGS: Between April 23 and Nov 4, 2020, 23 848 participants were enrolled and 11 636 participants (7548 in the UK, 4088 in Brazil) were included in the interim primary efficacy analysis. In participants who received two standard doses, vaccine efficacy was 62·1% (95% CI 41·0-75·7; 27 [0·6%] of 4440 in the ChAdOx1 nCoV-19 group vs71 [1·6%] of 4455 in the control group) and in participants who received a low dose followed by a standard dose, efficacy was 90·0% (67·4-97·0; three [0·2%] of 1367 vs 30 [2·2%] of 1374; pinteraction=0·010). Overall vaccine efficacy across both groups was 70·4% (95·8% CI 54·8-80·6; 30 [0·5%] of 5807 vs 101 [1·7%] of 5829). From 21 days after the first dose, there were ten cases hospitalised for COVID-19, all in the control arm; two were classified as severe COVID-19, including one death. There were 74 341 person-months of safety follow-up (median 3·4 months, IQR 1·3-4·8): 175 severe adverse events occurred in 168 participants, 84 events in the ChAdOx1 nCoV-19 group and 91 in the control group. Three events were classified as possibly related to a vaccine: one in the ChAdOx1 nCoV-19 group, one in the control group, and one in a participant who remains masked to group allocation. INTERPRETATION: ChAdOx1 nCoV-19 has an acceptable safety profile and has been found to be efficacious against symptomatic COVID-19 in this interim analysis of ongoing clinical trials. FUNDING: UK Research and Innovation, National Institutes for Health Research (NIHR), Coalition for Epidemic Preparedness Innovations, Bill & Melinda Gates Foundation, Lemann Foundation, Rede D'Or, Brava and Telles Foundation, NIHR Oxford Biomedical Research Centre, Thames Valley and South Midland's NIHR Clinical Research Network, and AstraZeneca

Safety and efficacy of the ChAdOx1 nCoV-19 vaccine (AZD1222) against SARS-CoV-2: an interim analysis of four randomised controlled trials in Brazil, South Africa, and the UK

Background A safe and efficacious vaccine against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), if deployed with high coverage, could contribute to the control of the COVID-19 pandemic. We evaluated the safety and efficacy of the ChAdOx1 nCoV-19 vaccine in a pooled interim analysis of four trials. Methods This analysis includes data from four ongoing blinded, randomised, controlled trials done across the UK, Brazil, and South Africa. Participants aged 18 years and older were randomly assigned (1:1) to ChAdOx1 nCoV-19 vaccine or control (meningococcal group A, C, W, and Y conjugate vaccine or saline). Participants in the ChAdOx1 nCoV-19 group received two doses containing 5 × 1010 viral particles (standard dose; SD/SD cohort); a subset in the UK trial received a half dose as their first dose (low dose) and a standard dose as their second dose (LD/SD cohort). The primary efficacy analysis included symptomatic COVID-19 in seronegative participants with a nucleic acid amplification test-positive swab more than 14 days after a second dose of vaccine. Participants were analysed according to treatment received, with data cutoff on Nov 4, 2020. Vaccine efficacy was calculated as 1 - relative risk derived from a robust Poisson regression model adjusted for age. Studies are registered at ISRCTN89951424 and ClinicalTrials.gov, NCT04324606, NCT04400838, and NCT04444674. Findings Between April 23 and Nov 4, 2020, 23 848 participants were enrolled and 11 636 participants (7548 in the UK, 4088 in Brazil) were included in the interim primary efficacy analysis. In participants who received two standard doses, vaccine efficacy was 62·1% (95% CI 41·0–75·7; 27 [0·6%] of 4440 in the ChAdOx1 nCoV-19 group vs71 [1·6%] of 4455 in the control group) and in participants who received a low dose followed by a standard dose, efficacy was 90·0% (67·4–97·0; three [0·2%] of 1367 vs 30 [2·2%] of 1374; pinteraction=0·010). Overall vaccine efficacy across both groups was 70·4% (95·8% CI 54·8–80·6; 30 [0·5%] of 5807 vs 101 [1·7%] of 5829). From 21 days after the first dose, there were ten cases hospitalised for COVID-19, all in the control arm; two were classified as severe COVID-19, including one death. There were 74 341 person-months of safety follow-up (median 3·4 months, IQR 1·3–4·8): 175 severe adverse events occurred in 168 participants, 84 events in the ChAdOx1 nCoV-19 group and 91 in the control group. Three events were classified as possibly related to a vaccine: one in the ChAdOx1 nCoV-19 group, one in the control group, and one in a participant who remains masked to group allocation. Interpretation ChAdOx1 nCoV-19 has an acceptable safety profile and has been found to be efficacious against symptomatic COVID-19 in this interim analysis of ongoing clinical trials

Lutein Attenuates Both Apoptosis and Autophagy upon Cobalt (II) Chloride-Induced Hypoxia in Rat Műller Cells

<div><p>Retinal ischemia/reperfusion injury is a common feature of various retinal diseases such as glaucoma and diabetic retinopathy. Lutein, a potent anti-oxidant, is used to improve visual function in patients with age-related macular degeneration (AMD). Lutein attenuates apoptosis, oxidative stress and inflammation in animal models of acute retinal ischemia/hypoxia. Here, we further show that lutein improved Műller cell viability and enhanced cell survival upon hypoxia-induced cell death through regulation of intrinsic apoptotic pathway. Moreover, autophagy was activated upon treatment of cobalt (II) chloride, indicating that hypoxic injury not only triggered apoptosis but also autophagy in our <i>in vitro</i> model. Most importantly, we report for the first time that lutein treatment suppressed autophagosome formation after hypoxic insult and lutein administration could inhibit autophagic event after activation of autophagy by a pharmacological approach (rapamycin). Taken together, lutein may have a beneficial role in enhancing glial cell survival after hypoxic injury through regulating both apoptosis and autophagy.</p></div

Anti-autophagic property of lutein was involved in mTOR-mediated autophagy pathway.

<p><b>(A)</b> Western blotting and the densitometric analysis showed that the phosphorylated AMPK was up-regulated in CoCl₂-treated cells. <b>(B-D)</b> Protein levels of mTOR-associated proteins including P-mTOR, P-p70S6K and P-ULK1 (Ser757) were measured by Western blotting (normalized by β-actin) and quantified by densitometry. 20μM of lutein was able to restore the phosphorylation levels of P-mTOR and P-p70S6K upon CoCl₂-induced hypoxia. (E) Rapamycin was used to induce autophagy and chloroquine was also added to block the formation of autolysosome upon rapamycin-mediated autophagy. Densitometry analysis showed that LC3II protein expression was up-regulated in rapamycin-induced autophagy and accumulated in the presence of chloroquine (lane 6). Lutein treatment was able to decrease LC3II expression in rMC-1 cells upon rapamycin and chloroquine co-treatment (lane 7). n = 5 in each group. ^*<i>P</i>< 0.05, ^**<i>P<0</i>.<i>01</i>, ^***<i>P</i><0.001 versus normal control group; ^#<i>P</i>< 0.05, ^##<i>P</i><0.01 versus Lutein-treated group; ^#<i>P</i>< 0.05, rapamycin and chloroquine co-treatment group.</p

The length of lutein treatment in rMC-1 cells upon CoCl₂-induced hypoxia.

<p>rMC-1 cells were exposed to CoCl₂ (300μM) with or without lutein (20μM) for different periods of time. <b>(A-D)</b> Protein levels of different apoptotic-related proteins including Bcl-2, Bcl-X_L, Bax, and cleaved caspase 3 were measured by Western blotting (normalized by β-actin) and quantified by densitometry. Lutein treatment was able to rescue cells upon CoCl₂-induced cell death by up-regulating protein expression of Bcl-2 and suppressing cleaved caspase 3 at 24 hours. <b>(E)</b> Densitometry analysis of ratio of Bax and Bcl-2 protein expression at different time points. Lutein improved rMC-1 cell survival by decreasing Bax/Bcl-2 ratio at 24 hours. n = 5 in each group. ^*<i>P</i><0.05, ^**<i>P</i>< 0.01, ^***<i>P</i><0.001 versus normal control group; ^#<i>P</i>< 0.05, ^###<i>P</i><0.001 versus vehicle-treated group.</p

Lutein rescued rMC-1 cells from CoCl₂-induced hypoxic injury.

<p>rMC-1 cells were exposed to CoCl₂ (300μM) with or without lutein for various periods. Representative photographs of rMC-1 cells <b>(A-C)</b> normal control, <b>(D-F)</b> hypoxia with vehicle (0.01% DMSO), <b>(G-I)</b> hypoxia with lutein (20μM). <b>(J)</b> Percentage of cell viability. Treatment of lutein only without hypoxia did not affect the viability when compared with the normal control. Lutein-treated rMC-1 cells showed higher cell viability when compared with the vehicle-treated group at 24 hours. <b>(K)</b> Percentage of lactate dehydrogenase (LDH) release from damaged cells. Lutein attenuated LDH release after CoCl₂-induced injury when compared with that in vehicle-treated group at 24 hours. n = 5 in each group. ^**<i>P</i>< 0.01, ^***<i>P</i><0.001 versus normal control group; ^#<i>P</i>< 0.05, ^##<i>P</i><0.01 versus vehicle-treated group. Scale bar, 100 μm.</p

Number of apoptotic nuclei was attenuated in lutein-treated cells.

<p>Apoptotic nuclei were revealed by TUNEL assay after CoCl₂ treatment at different time points. <b>(A)</b> Representative images of TUNEL-positive nuclei (green) and DAPI-stained nuclei (blue) in rMC-1 cells. <b>(B)</b> Quantification of TUNEL-positive cells. n = 5 in each group. Lutein administration significantly reduced the number of TUNEL-stained nuclei. Scale bar, 100 μm. ^*<i>P</i><0.05, ^**<i>P</i>< 0.01, ^***<i>P</i><0.001 versus normal control group; ^#<i>P</i>< 0.05 versus vehicle-treated group.</p

Anti-apoptotic effects of lutein in rMC-1 cells upon CoCl₂-induced hypoxia.

<p>rMC-1 cells were exposed to CoCl₂ (300μM) with various concentration of lutein for 24 hours. <b>(A-D)</b> Protein levels of different apoptotic-related proteins including Bcl-2, Bcl-X_L, Bax, and cleaved caspase 3 were measured by Western blotting (normalized by β-actin) and quantified by densitometry. 20μM of Lutein was able to up-regulate protein expression of Bcl-2 and protein level of cleaved caspase 3 was inhibited by both 10μM and 20μM of lutein. <b>(E)</b> Densitometry analysis of ratio of Bax and Bcl-2 protein expression with different concentration of lutein. Lutein improved rMC-1 cell survival by decreasing Bax/Bcl-2 ratio. n = 5 in each group. ^*<i>P</i><0.05, ^***<i>P</i><0.001 versus normal control group; ^#<i>P</i>< 0.05, ^##<i>P</i><0.01 versus vehicle-treated group. N, Normal Control; Veh, Vehicle (0.01% DMSO).</p