COVID-19 was declared a pandemic on March 11 by WHO, due to its great threat to global public health. The coronavirus main protease (M
, also called 3CLpro) is essential for processing and maturation ...of the viral polyprotein, therefore recognized as an attractive drug target. Here we show that a clinically approved anti-HCV drug, Boceprevir, and a pre-clinical inhibitor against feline infectious peritonitis (corona) virus (FIPV), GC376, both efficaciously inhibit SARS-CoV-2 in Vero cells by targeting M
. Moreover, combined application of GC376 with Remdesivir, a nucleotide analogue that inhibits viral RNA dependent RNA polymerase (RdRp), results in sterilizing additive effect. Further structural analysis reveals binding of both inhibitors to the catalytically active side of SARS-CoV-2 protease M
as main mechanism of inhibition. Our findings may provide critical information for the optimization and design of more potent inhibitors against the emerging SARS-CoV-2 virus.
SARS-CoV-2, a β-coronavirus, has rapidly spread across the world, highlighting its high transmissibility, but the underlying morphogenesis and pathogenesis remain poorly understood. Here, we ...characterize the replication dynamics, cell tropism and morphogenesis of SARS-CoV-2 in organotypic human airway epithelial (HAE) cultures. SARS-CoV-2 replicates efficiently and infects both ciliated and secretory cells in HAE cultures. In comparison, HCoV-NL63 replicates to lower titers and is only detected in ciliated cells. SARS-CoV-2 shows a similar morphogenetic process as other coronaviruses but causes plaque-like cytopathic effects in HAE cultures. Cell fusion, apoptosis, destruction of epithelium integrity, cilium shrinking and beaded changes are observed in the plaque regions. Taken together, our results provide important insights into SARS-CoV-2 cell tropism, replication and morphogenesis.
As one paper in a series reporting on a large reverberation mapping campaign of super-Eddington accreting massive black holes (SEAMBHs) in active galactic nuclei (AGNs), we present the results of 10 ...SEAMBHs monitored spectroscopically during 2015-2017. Six of them are observed for the first time, and have generally higher 5100 luminosities than the SEAMBHs monitored in our campaign from 2012 to 2015; the remaining four are repeat observations to check if their previous lags change. Similar to the previous SEAMBHs, the Hβ time lags of the newly observed objects are shorter than the values predicted by the canonical RHβ-L5100 relation of sub-Eddington AGNs, by factors of ∼2-6, depending on the accretion rate. The four previously observed objects have lags consistent with previous measurements. We provide linear regressions for the RHβ-L5100 relation, solely for the SEAMBH sample and for low-accretion AGNs. We find that the relative strength of Fe ii and the profile of the Hβ emission line can be used as proxies of accretion rate, showing that the shortening of Hβ lags depends on accretion rates. The recent SDSS-RM discovery of shortened Hβ lags in AGNs with low accretion rates provides compelling evidence for retrograde accretion onto the black hole. These evidences show that the canonical RHβ-L5100 relation holds only in AGNs with moderate accretion rates. At low accretion rates, it should be revised to include the effects of black hole spin, whereas the accretion rate itself becomes a key factor in the regime of high accretion rates.
Abstract
Background
The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) first broke out in 2019 and subsequently spread worldwide. Chloroquine has been sporadically used in treating ...SARS-CoV-2 infection. Hydroxychloroquine shares the same mechanism of action as chloroquine, but its more tolerable safety profile makes it the preferred drug to treat malaria and autoimmune conditions. We propose that the immunomodulatory effect of hydroxychloroquine also may be useful in controlling the cytokine storm that occurs late phase in critically ill patients with SARS-CoV-2. Currently, there is no evidence to support the use of hydroxychloroquine in SARS-CoV-2 infection.
Methods
The pharmacological activity of chloroquine and hydroxychloroquine was tested using SARS-CoV-2–infected Vero cells. Physiologically based pharmacokinetic (PBPK) models were implemented for both drugs separately by integrating their in vitro data. Using the PBPK models, hydroxychloroquine concentrations in lung fluid were simulated under 5 different dosing regimens to explore the most effective regimen while considering the drug’s safety profile.
Results
Hydroxychloroquine (EC50 = 0.72 μM) was found to be more potent than chloroquine (EC50 = 5.47 μM) in vitro. Based on PBPK models results, a loading dose of 400 mg twice daily of hydroxychloroquine sulfate given orally, followed by a maintenance dose of 200 mg given twice daily for 4 days is recommended for SARS-CoV-2 infection, as it reached 3 times the potency of chloroquine phosphate when given 500 mg twice daily 5 days in advance.
Conclusions
Hydroxychloroquine was found to be more potent than chloroquine to inhibit SARS-CoV-2 in vitro.
Hydroxychloroquine was found to be more potent than chloroquine at inhibiting SARS-CoV-2 in vitro. Hydroxychloroquine sulfate 400 mg given twice daily for 1 day, followed by 200 mg twice daily for 4 more days, is recommended to treat SARS-CoV-2 infection.
Different analytical methods including Fourier transform infrared (FTIR), atomic force microscopy (AFM), zeta potential measurements, contact angle measurements and spontaneous imbibition tests were ...utilized to make clear the mechanism for wettability alteration of oil-wet sandstone surface using different surfactants. Results show that among three types of surfactants including cationic surfactants, anionic surfactants and nonionic surfactants, the cationic surfactant CTAB demonstrates the best effect on the wettability alteration of oil-wet sandstone surface. The positively charged head groups of CTAB molecules and carboxylic acid groups from crude oil could interact to form ion pairs, which could be desorbed from the solid surface and solubilized into the micelle formed by CTAB. Thus, the water-wetness of the solid surface is improved. Nonionic surfactant TX-100 could be adsorbed on oil-wet sandstone surface through hydrogen bonds and hydrophobic interaction to alter the wettability of oil-wet solid surface. The wettability alteration of oil-wet sandstone surface using the anionic surfactant POE(1) is caused by hydrophobic interaction. Due to the electrostatic repulsion between the anionic surfactant and the negatively charged surface, POE(1) shows less effect on the wettability alteration of oil-wet sandstone surface.
This is the eighth in a series of papers reporting on a large reverberation mapping (RM) campaign to measure black hole (BH) mass in active galactic nuclei with high accretion rates. We employ the ...recently developed dynamical modeling approach for broad-line regions (BLRs) based on the method of Pancoast et al. to analyze the RM data set of Mrk 142 observed in the first monitoring season. In this approach, continuum variations are reconstructed using a damped random walk process, and BLR structure is delineated using a flexible disk-like geometry, in which BLR clouds move around the central BH with Keplerian orbits or inflow/outflow motion. The approach also includes the possibilities of anisotropic emission from BLR clouds, nonlinear response of the line emission to the continuum, and different long-term trends in the continuum and emission-line variations. We implement the approach in a Bayesian framework that is apt for parallel computation and use a Markov chain Monte Carlo technique to recover the parameters and uncertainties for the modeling, including the mass of the central BH. We apply three BLR models with different prescriptions of BLR cloud distributions and find that the best model for fitting the data of Mrk 142 is a two-zone BLR model, consistent with the theoretical BLR model surrounding slim accretion disks. The best model yields a BH mass of , resulting in a virial factor of for the full width at half maximum of the Hβ line measured from the mean spectrum. The virial factors for the other measures of the Hβ line width are also presented.
Soaring cases of coronavirus disease (COVID-19) are pummeling the global health system. Overwhelmed health facilities have endeavored to mitigate the pandemic, but mortality of COVID-19 continues to ...increase. Here, we present a mortality risk prediction model for COVID-19 (MRPMC) that uses patients' clinical data on admission to stratify patients by mortality risk, which enables prediction of physiological deterioration and death up to 20 days in advance. This ensemble model is built using four machine learning methods including Logistic Regression, Support Vector Machine, Gradient Boosted Decision Tree, and Neural Network. We validate MRPMC in an internal validation cohort and two external validation cohorts, where it achieves an AUC of 0.9621 (95% CI: 0.9464-0.9778), 0.9760 (0.9613-0.9906), and 0.9246 (0.8763-0.9729), respectively. This model enables expeditious and accurate mortality risk stratification of patients with COVID-19, and potentially facilitates more responsive health systems that are conducive to high risk COVID-19 patients.
We study the long-term variability in the optical monitoring database of Ark 120, a nearby radio-quiet active galactic nucleus (AGN) at a distance of 143 Mpc (z = 0.03271). We compiled the historical ...archival photometric and spectroscopic data since 1974 and conducted a new two-year monitoring campaign in 2015-2017, resulting in a total temporal baseline over four decades. The long-term variations in the optical continuum exhibit a wave-like pattern and the Hβ integrated flux series varies with a similar behavior. The broad Hβ profiles have asymmetric double peaks, which change strongly with time and tend to merge into a single peak during some epochs. The period in the optical continuum determined from various period-search methods is about 20 yr, and the estimated false alarm probability with null hypothesis simulations is about 1 × 10−3. The overall variations of the broad Hβ profiles also follow the same period. However, the present database only covers two cycles of the suggested period, which strongly encourages continued monitoring to track more cycles and confirm the periodicity. Nevertheless, in light of the possible periodicity and the complicated Hβ profile, Ark 120 is one candidate of the nearest radio-quiet AGNs with possible periodic variability, and it is thereby a potential candidate host for a sub-parsec supermassive black hole binary.
Human steroid 5α-reductase 2 (SRD5A2) is an integral membrane enzyme in steroid metabolism and catalyzes the reduction of testosterone to dihydrotestosterone. Mutations in the SRD5A2 gene have been ...linked to 5α-reductase deficiency and prostate cancer. Finasteride and dutasteride, as SRD5A2 inhibitors, are widely used antiandrogen drugs for benign prostate hyperplasia. The molecular mechanisms underlying enzyme catalysis and inhibition for SRD5A2 and other eukaryotic integral membrane steroid reductases remain elusive due to a lack of structural information. Here, we report a crystal structure of human SRD5A2 at 2.8 Å, revealing a unique 7-TM structural topology and an intermediate adduct of finasteride and NADPH as NADP-dihydrofinasteride in a largely enclosed binding cavity inside the transmembrane domain. Structural analysis together with computational and mutagenesis studies reveal the molecular mechanisms of the catalyzed reaction and of finasteride inhibition involving residues E57 and Y91. Molecular dynamics simulation results indicate high conformational dynamics of the cytosolic region that regulate NADPH/NADP
exchange. Mapping disease-causing mutations of SRD5A2 to our structure suggests molecular mechanisms for their pathological effects. Our results offer critical structural insights into the function of integral membrane steroid reductases and may facilitate drug development.
Coronaviruses (CoVs) act as cross-species viruses and have the potential to spread rapidly into new host species and cause epidemic diseases. Despite the severe public health threat of severe acute ...respiratory syndrome coronavirus and Middle East respiratory syndrome CoV (MERS-CoV), there are currently no drugs available for their treatment; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are urgently needed. To search for effective inhibitory agents, we performed high-throughput screening (HTS) of a 2,000-compound library of approved drugs and pharmacologically active compounds using the established genetically engineered human CoV OC43 (HCoV-OC43) strain expressing
luciferase (rOC43-ns2Del-Rluc) and validated the inhibitors using multiple genetically distinct CoVs
We screened 56 hits from the HTS data and validated 36 compounds
using wild-type HCoV-OC43. Furthermore, we identified seven compounds (lycorine, emetine, monensin sodium, mycophenolate mofetil, mycophenolic acid, phenazopyridine, and pyrvinium pamoate) as broad-spectrum inhibitors according to their strong inhibition of replication by four CoVs
at low-micromolar concentrations. Additionally, we found that emetine blocked MERS-CoV entry according to pseudovirus entry assays and that lycorine protected BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This represents the first demonstration of
real-time bioluminescence imaging to monitor the effect of lycorine on the spread and distribution of HCoV-OC43 in a mouse model. These results offer critical information supporting the development of an effective therapeutic strategy against CoV infection.
Currently, there is no approved therapy to treat coronavirus infection; therefore, broad-spectrum inhibitors of emerging and endemic CoVs are needed. Based on our high-throughput screening assay using a compound library, we identified seven compounds with broad-spectrum efficacy against the replication of four CoVs
Additionally, one compound (lycorine) was found to protect BALB/c mice against HCoV-OC43-induced lethality by decreasing viral load in the central nervous system. This inhibitor might offer promising therapeutic possibilities for combatting novel CoV infections in the future.
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