Structure-Based Identification of Natural Products as SARS-CoV-2 Mpro Antagonist from Echinacea angustifolia Using Computational Approaches

Coronavirus disease-19 (COVID-19) pandemic, caused by the novel SARS-CoV-2 virus, continues to be a global threat. The number of cases and deaths will remain escalating due to the lack of effective therapeutic agents. Several studies have established the importance of the viral main protease (M^pro) in the replication of SARS-CoV-2 which makes it an attractive target for antiviral drug development, including pharmaceutical repurposing and other medicinal chemistry approaches. Identification of natural products with considerable inhibitory potential against SARS-CoV-2 could be beneficial as a rapid and potent alternative with drug-likeness by comparison to de novo antiviral drug discovery approaches. Thereof, we carried out the structure-based screening of natural products from Echinacea-angustifolia, commonly used to prevent cold and other microbial respiratory infections, targeting SARS-CoV-2 M^pro. Four natural products namely, Echinacoside, Quercetagetin 7-glucoside, Levan N, Inulin from chicory, and 1,3-Dicaffeoylquinic acid, revealed significant docking energy (>−10 kcal/mol) in the SARS-CoV-2 M^pro catalytic pocket via substantial intermolecular contacts formation against co-crystallized ligand (<−4 kcal/mol). Furthermore, the docked poses of SARS-CoV-2 M^pro with selected natural products showed conformational stability through molecular dynamics. Exploring the end-point net binding energy exhibited substantial contribution of Coulomb and van der Waals interactions to the stability of respective docked conformations. These results advocated the natural products from Echinacea angustifolia for further experimental studies with an elevated probability to discover the potent SARS-CoV-2 M^pro antagonist with higher affinity and drug-likeness.     

Preparation of Nitrogen-doped Holey Multilayer Graphene Using High-Energy Ball Milling of Graphite in Presence of Melamine

Holey graphene, consisting of graphene sheets with in-plane nanopores, has recently attracted more attention as it expands graphene applications to other fields inaccessible by the pristine graphene. To ensure an effective implementation of holey graphene in the market, it is crucial to explore new preparation methods that are simple, cost effective, eco-friendly, versatile, and scalable. While ball milling of graphite in presence of exfoliating agents was found very effective in the preparation of graphene (doped and undoped) and graphene-composites, this technique remains unexplored for the preparation of holey graphene. In the present work, Nitrogen-doped multilayer holey graphene sheets were prepared by an all-solid, one-step procedure based on high-energy ball milling of graphite as the starting material in presence of melamine in a shaker-type mill for 1 hour under ambient conditions. Melamine acted simultaneously as an exfoliating agent to enhance the exfoliation of graphene layers and a diluent to protect graphite against the continuous fragmentation into amorphous carbon during the high-energy “shock” mode of ball milling. The high-energy “shock” mode of ball milling of graphite in presence of melamine induced the formation of multilayer defective graphene as an intermediate product before being converted into N-doped multilayer holey graphene after the removal of the in-plane defects during the milling process. The characterization of the final product confirmed the formation of N-doped multilayer holey graphene with a content in nitrogen as high as 12.96 at.%, making it promising for energy storage and energy conversion applications.     

Aerobic activity before and following short-duration static stretching improves range of motion and performance vs. a traditional warm-up

Many activities necessitate a high degree of static joint range of motion (ROM) for an extended duration. The objective of this study was to examine whether ROM could be improved with a short duration and volume of static stretching within a warm-up, without negatively impacting performance. Ten male recreationally active participants completed 2 separate protocols to examine changes in ROM and performance, respectively, with different warm-ups. The warm-up conditions for the ROM protocol were static stretching (SS), consisting of 6 repetitions of 6 s stretches; 10?min of running prior to the SS (AS); and 5 min of running before and after the SS (ASA). The performance protocol included a control condition of 10?min of running. Measures for the ROM protocol included hip flexion ROM, passive leg extensor tension, and hamstring electromyographic (EMG) activity at pre-warm-up, and at 1, 10, 20, and 30?min post-warm-up. Performance measures included countermovement jump (CMJ) height, reaction time (RT), movement time (MT), and balance at pre-warm-up and at 1 and 10?min post-warm-up. The ASA produced greater ROM overall than the SS and AS conditions (p?< 0.0001), persisting for 30?min. There were no significant alterations in passive muscle tension or EMG. For the performance protocol, there were no main effects for condition, but there was a main effect for time, with CMJ height being greater at 1 and 10?min post-warm-up (p = 0.0004). Balance ratios and MT improved at 10?min post-warm-up (p?< 0.0001). Results indicate that the ASA method can provide ROM improvements for 30?min with either facilitation or no impairment in performance. This may be especially important for athletes who substitute later into a game with minimal time for a full warm-up.     

The basic C-terminal amino acids of calcium-binding protein S100A4 promote metastasis

The calcium-binding protein S100A4 can induce a metastatic phenotype in animal model systems and its expression in various human cancers has been shown to be associated with metastasis and reduced patient survival. Using a series of nested deletion mutants, it is now shown that the two C-terminal lysine residues are required for the enhanced metastasis, invasion and migration abilities that S100A4 confers on cells in a model system of metastasis. Basic C-terminal residues enhance the affinity between S100A4 and its best characterized target, a recombinant C-terminal fragment of non-muscle myosin II heavy chain isoform A (NMMHC-IIA). In wild-type S100A4 protein, the presence of the C-terminal lysine, residue 101, enhances the rate of association between S100A4 and NMMHC-IIA. These results identify the amino acids of S100A4 that are involved in metastasis induction and show that the C-terminal region of S100A4 is a possible target for inhibitors of its metastatic action.     

Apremilast reversed carfilzomib-induced cardiotoxicity through inhibition of oxidative stress,NF-κB and MAPK signaling in rats

Carfilzomib (CFZ), is a potent, selective second generation proteasome inhibitor, used for the treatment of multiple myeloma. The aim of the present study was to investigate the possible protective effect of apremilast (AP) on the CFZ -induced cardiotoxicity. Rats were randomly divided into four groups: Group 1, served as the control group, received normal saline. Group 2, served as the toxic group, received CFZ (4?mg/kg, intraperitoneally [i.p.]). Groups 3 and 4, served as treatment groups, and received CFZ with concomitant oral administration of AP in doses of 10 and 20?mg/kg/day, respectively. In the present study, administration of CFZ resulted in a significant increase in serum aspartate transaminase (AST), lactate dehydrogenase (LDH), creatine kinase (CK) and creatine kinase-MB (CK-MB), which were reversed by treatment with AP. CFZ resulted in a significant increase in heart malondialdehyde (MDA) contents and decrease in cardiac glutathione (GSH) level and catalase (CAT) enzyme activity which were significantly reversed by treatment with AP. Induction of cardiotoxicity by CFZ significantly increased caspase-3 enzyme activity which were reversed by treatment with AP. RT-PCR analysis revealed an increased mRNA expression of NF-κB, ERK and JNK which were reversed by treatment with AP in cardiac tissues. Western blot analysis revealed an increased expression of caspase-3 and NF-κB p65 and a decrease expression of inhibitory kappa B-alpha (Iκbα) with CFZ, which were reversed by treatment with AP. In conclusion, apremilast showed protective effect against CFZ-induced cardiotoxicity.     

Metformin attenuates V-domain Ig suppressor of T-cell activation through the aryl hydrocarbon receptor pathway in Melanoma: In Vivo and In Vitro Studies

Melanoma is an aggressive skin cancer with a high rate of metastasis to other organs. Recent studies specified the overexpression of V-domain Ig suppressor of T-cell activation (VISTA) and Aryl Hydrocarbon Receptor (AHR) in melanoma. Metformin shows anti-tumor activities in several cancer types. However, the mechanism is unclear. This study aims to investigate the inhibitory effect of metformin on VISTA via AHR in melanoma cells (CHL-1, B16) and animal models. VISTA and AHR levels were assessed by qPCR, Western blot, immunofluorescence microscope, flow cytometry, and immunohistochemistry. Here, metformin significantly decreased VISTA and AHR levels in vitro and in vivo. Furthermore, metformin inhibited all AHR-regulated genes. VISTA levels were dramatically inhibited by AHR modulations using shRNA and αNF, confirming the central role of AHR in VISTA. Finally, melanoma cells were xenografted in C57BL/6 and nude mice. Metformin significantly reduced the tumor volume and growth rate. Likewise, VISTA and AHR-regulated protein levels were suppressed in both models. These findings demonstrate for the first time that VISTA is suppressed by metformin and identified a new regulatory mechanism through AHR. The data suggest that metformin could be a new potential therapeutic strategy to treat melanoma patients combined with targeted immune checkpoint inhibitors.