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1.
Huan Jing Ke Xue ; 44(5): 2430-2440, 2023 May 08.
Article in Chinese | MEDLINE | ID: covidwho-20237414

ABSTRACT

To investigate the change characteristics of secondary inorganic ions in PM2.5 at different pollution stages before and after COVID-19, the online monitoring of winter meteorological and atmospheric pollutant concentrations in Zhengzhou from December 15, 2019 to February 15, 2020 was conducted using a high-resolution (1 h) online instrument. This study analyzed the causes of the haze process of COVID-19, the diurnal variation characteristics of air pollutants, and the distribution characteristics of air pollutants at different stages of haze.The results showed that Zhengzhou was mainly controlled by the high-pressure ridge during the haze process, and the weather situation was stable, which was conducive to the accumulation of air pollutants. SNA was the main component of water-soluble ions, accounting for more than 90%. Home isolation measures during COVID-19 had different impacts on the distribution characteristics of air pollutants in different haze stages. After COVID-19, the concentration of PM2.5 in the clean, occurrence, and dissipation stages increased compared with that before COVID-19 but significantly decreased in the development stage. The home isolation policy significantly reduced the high value of PM2.5. The concentrations of NO2, SO2, NH3, and CO were the highest in the haze development stage, showing a trend of first increasing and then decreasing. The concentration of O3 was lowest in the pre-COVID-19 development stage but highest in the post-COVID-19 development stage. The linear correlation between[NH4+]/[SO42-] and[NO3-]/[SO42-] at different time periods before and after COVID-19 was strong, indicating that the home isolation policy of COVID-19 did not change the generation mode of NO3-, and the corresponding reaction was always the main generation mode of NO3-. The correlation between[excess-NH4+] and[NO3-] was high in different periods before COVID-19, and NO3- generation was related to the increase in NH3 or NH4+ in the process of PM2.5 pollution in Zhengzhou.


Subject(s)
Air Pollutants , Air Pollution , COVID-19 , Humans , Particulate Matter/analysis , Environmental Monitoring/methods , COVID-19/epidemiology , Respiratory Aerosols and Droplets , Air Pollutants/analysis , Air Pollution/analysis , Ions/analysis , Seasons , China/epidemiology
2.
Acc Chem Res ; 56(12): 1458-1468, 2023 06 20.
Article in English | MEDLINE | ID: covidwho-20234847

ABSTRACT

Native mass spectrometry is nowadays widely used for determining the mass of intact proteins and their noncovalent biomolecular assemblies. While this technology performs well in the mass determination of monodisperse protein assemblies, more real-life heterogeneous protein complexes can pose a significant challenge. Factors such as co-occurring stoichiometries, subcomplexes, and/or post-translational modifications, may especially hamper mass analysis by obfuscating the charge state inferencing that is fundamental to the technique. Moreover, these mass analyses typically require measurement of several million molecules to generate an analyzable mass spectrum, limiting its sensitivity. In 2012, we introduced an Orbitrap-based mass analyzer with extended mass range (EMR) and demonstrated that it could be used to obtain not only high-resolution mass spectra of large protein macromolecular assemblies, but we also showed that single ions generated from these assemblies provided sufficient image current to induce a measurable charge-related signal. Based on these observations, we and others further optimized the experimental conditions necessary for single ion measurements, which led in 2020 to the introduction of single-molecule Orbitrap-based charge detection mass spectrometry (Orbitrap-based CDMS). The introduction of these single molecule approaches has led to the fruition of various innovative lines of research. For example, tracking the behavior of individual macromolecular ions inside the Orbitrap mass analyzer provides unique, fundamental insights into mechanisms of ion dephasing and demonstrated the (astonishingly high) stability of high mass ions. Such fundamental information will help to further optimize the Orbitrap mass analyzer. As another example, the circumvention of traditional charge state inferencing enables Orbitrap-based CDMS to extract mass information from even extremely heterogeneous proteins and protein assemblies (e.g., glycoprotein assemblies, cargo-containing nanoparticles) via single molecule detection, reaching beyond the capabilities of earlier approaches. We so far demonstrated the power of Orbitrap-based CDMS applied to a variety of fascinating systems, assessing for instance the cargo load of recombinant AAV-based gene delivery vectors, the buildup of immune-complexes involved in complement activation, and quite accurate masses of highly glycosylated proteins, such as the SARS-CoV-2 spike trimer proteins. With such widespread applications, the next objective is to make Orbitrap-based CDMS more mainstream, whereby we still will seek to further advance the boundaries in sensitivity and mass resolving power.


Subject(s)
COVID-19 , Humans , SARS-CoV-2 , Mass Spectrometry/methods , Proteins/chemistry , Ions , Macromolecular Substances/chemistry
3.
J Biol Chem ; 299(6): 104787, 2023 06.
Article in English | MEDLINE | ID: covidwho-2308970

ABSTRACT

Understanding the functional properties of severe acute respiratory syndrome coronavirus 2 nonstructural proteins is essential for defining their roles in the viral life cycle, developing improved therapeutics and diagnostics, and countering future variants. Coronavirus nonstructural protein Nsp15 is a hexameric U-specific endonuclease whose functions, substrate specificity, mechanism, and dynamics are not fully defined. Previous studies report that Nsp15 requires Mn2+ ions for optimal activity; however, the effects of divalent ions on Nsp15 reaction kinetics have not been investigated in detail. Here, we analyzed the single- and multiple-turnover kinetics for model ssRNA substrates. Our data confirm that divalent ions are dispensable for catalysis and show that Mn2+ activates Nsp15 cleavage of two different ssRNA oligonucleotide substrates but not a dinucleotide. Biphasic kinetics of ssRNA substrates demonstrates that Mn2+ stabilizes alternative enzyme states that have faster substrate cleavage on the enzyme. However, we did not detect Mn2+-induced conformational changes using CD and fluorescence spectroscopy. The pH-rate profiles in the presence and absence of Mn2+ reveal active-site ionizable groups with similar pKas of ca. 4.8 to 5.2. An Rp stereoisomer phosphorothioate modification at the scissile phosphate had minimal effect on catalysis supporting a mechanism involving an anionic transition state. However, the Sp stereoisomer is inactive because of weak binding, consistent with models that position the nonbridging phosphoryl oxygen deep in the active site. Together, these data demonstrate that Nsp15 employs a conventional acid-base catalytic mechanism passing through an anionic transition state, and that divalent ion activation is substrate dependent.


Subject(s)
Endonucleases , Ions , RNA Cleavage , SARS-CoV-2 , Catalysis , COVID-19/microbiology , Endonucleases/genetics , Endonucleases/metabolism , Kinetics , Metals/chemistry , RNA Cleavage/genetics , SARS-CoV-2/enzymology , Ions/metabolism , Enzyme Activation , Manganese/chemistry , Hydrogen-Ion Concentration , Animals , Mice , Escherichia coli/genetics
4.
Commun Biol ; 6(1): 233, 2023 03 02.
Article in English | MEDLINE | ID: covidwho-2283223

ABSTRACT

Sensitive and rapid point-of-care assays have been crucial in the global response to SARS-CoV-2. Loop-mediated isothermal amplification (LAMP) has emerged as an important diagnostic tool given its simplicity and minimal equipment requirements, although limitations exist regarding sensitivity and the methods used to detect reaction products. We describe the development of Vivid COVID-19 LAMP, which leverages a metallochromic detection system utilizing zinc ions and a zinc sensor, 5-Br-PAPS, to circumvent the limitations of classic detection systems dependent on pH indicators or magnesium chelators. We make important strides in improving RT-LAMP sensitivity by establishing principles for using LNA-modified LAMP primers, multiplexing, and conducting extensive optimizations of reaction parameters. To enable point-of-care testing, we introduce a rapid sample inactivation procedure without RNA extraction that is compatible with self-collected, non-invasive gargle samples. Our quadruplexed assay (targeting E, N, ORF1a, and RdRP) reliably detects 1 RNA copy/µl of sample (=8 copies/reaction) from extracted RNA and 2 RNA copies/µl of sample (=16 copies/reaction) directly from gargle samples, making it one of the most sensitive RT-LAMP tests and even comparable to RT-qPCR. Additionally, we demonstrate a self-contained, mobile version of our assay in a variety of high-throughput field testing scenarios on nearly 9,000 crude gargle samples. Vivid COVID-19 LAMP can be an important asset for the endemic phase of COVID-19 as well as preparing for future pandemics.


Subject(s)
COVID-19 , Zinc , Humans , Colorimetry , COVID-19/diagnosis , SARS-CoV-2/genetics , DNA Primers , Ions
5.
Rapid Commun Mass Spectrom ; 37(8): e9481, 2023 Apr 30.
Article in English | MEDLINE | ID: covidwho-2219865

ABSTRACT

RATIONALE: The chemical constituents of Chinese patent medicine are usually different from those of crude medicine because of specific preparation processes. Chimonanthus nitens Oliv. leaf granule is widely used for prevention against COVID-19 in China. However, no research has been reported on the chemical constituents of the granule and their variation during the preparation process. METHODS: Fragmentation patterns of reference compounds were investigated using electrospray ionization mass spectrometry, and the new gas-phase reaction was demonstrated by electronic and steric effects and calculated chemistry. Then, a strategy based on new fragmentation patterns was used to profile aromatic constituents. In addition, based on untargeted metabolomics analytical workflow, a comparison was made on the chemical constituents of the leaf and granule. RESULTS: New fragmentation patterns related to two competing reactions, ring-opening and ring-closing reactions for coumarin, have been proposed and investigated in depth. The newly established diagnostic ion at m/z 81.0331 worked strongly in the assignment of OH-7 and substituent at C-8 of coumarin. McLafferty rearrangement occurring in coumarin glycoside while sugar group locating at C-4 was first observed, and new diagnostic ions at m/z 147.0440, 119.0488, and 91.0543 were constructed. CONCLUSIONS: Aromatic constituents of the granule were first profiled. A total of 114 aromatic compounds were identified; of these 85 compounds were identified first. Kaempferol-7-O-neohesperidoside and its homologues were mostly enriched in the granule. Considering their reported bioactivities, these analogues possibly contribute greatly to clinical efficacy. Our results provided a new fragmentation theory for coumarins and a new material basis for the quality control of the granule.


Subject(s)
COVID-19 , Drugs, Chinese Herbal , Spectrometry, Mass, Electrospray Ionization/methods , Drugs, Chinese Herbal/chemistry , Ions/chemistry , Plant Leaves/chemistry , Chromatography, High Pressure Liquid/methods
6.
J Hazard Mater ; 440: 129768, 2022 10 15.
Article in English | MEDLINE | ID: covidwho-2049471

ABSTRACT

Every two years, the Pollutant Toxic Ions and Molecules Conference, PTIM, meets the environmentalist, biologist, chemists and health researchers in Costa de Caparica, Portugal, to showcase the latest technologies, methodologies and research advances in pollution detection, contamination control, remediation, and related health issues, as well as policy implications.


Subject(s)
Environmental Pollutants , Environmental Restoration and Remediation , Environmental Pollutants/toxicity , Environmental Pollution , Ions , Pandemics
7.
J Phys Chem Lett ; 14(1): 88-94, 2023 Jan 12.
Article in English | MEDLINE | ID: covidwho-2185481

ABSTRACT

The receptor-binding domain of the SARS-CoV-2 spike mediates the key to binding the virus to the host receptor, but capturing the molecular signal of this spike RBD remains a formidable challenge. Here, we report a new surface-enhanced Raman spectroscopy (SERS) approach, which used gold nanoparticles prepared by low-speed constant-temperature centrifugation by bromine and calcium ions in two cleaning steps as the enhanced substrate to rapidly and accurately detect spike RBD large protein molecules in body fluids. The detection signal was extremely stable, and the orientation of the spike RBD on the enhanced substrate surface was also determined. This approach was specific in distinguishing different SARS-CoV-2 variants of spike RBD, including Delta, Beta, Gamma, and Omicron. Additionally, the enhanced substrate can identify biologically active or inactive spike RBD. This two-step cleaning enhanced substrate opens up opportunities not only for early diagnostics of SARS-CoV-2 virus but also for developing targeted drugs against viruses.


Subject(s)
Body Fluids , COVID-19 , Metal Nanoparticles , Humans , Bromides , COVID-19/diagnosis , Calcium , Gold , SARS-CoV-2 , Ions
8.
Int J Mol Sci ; 23(23)2022 Dec 02.
Article in English | MEDLINE | ID: covidwho-2143239

ABSTRACT

The ionic E-nanochannel (viroporin) is the weak point of SARS-CoV-2, the virus responsible for the (still threatening) COVID-19 since it is vital to the virus's budding and propagation. Therefore, targeting it to disable its functions ought to incapacitate, or at least weaken, the virus. The ionic currents inside this channel could be affected and disturbed by direct physical attack via the actions of external fields. The paper presents the first step towards the application of such methods in the fight against the current pandemic, numerical simulations of external fields' impact on ionic currents through viral channels. These simulations-based on the actual, detailed physical nanostructure of ionic channels, measured experimentally and reported in the literature-show that external physical fields can diminish the channel's currents and that the lower the channel's selectivity, the stronger the effect. Simulations suggest that SARS-CoV-2 E-viroporin is almost non-selective, which means that the whole virus ought to be highly vulnerable to the actions of external physical fields, much more vulnerable than the much more selective human cell ionic channels. If corroborated by experiment, this observation may result in an innovative method of dealing with the recent pandemic caused by SARS-CoV-2 and other similar viruses.


Subject(s)
COVID-19 , Frailty , Viruses , Humans , SARS-CoV-2 , Pandemics , Viroporin Proteins , Ion Channels , Ions
9.
Biochem J ; 479(20): 2175-2193, 2022 Oct 28.
Article in English | MEDLINE | ID: covidwho-2062282

ABSTRACT

Coronaviruses have been responsible for multiple challenging global pandemics, including coronavirus disease 2019 (COVID-19), which is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Papain-like protease (PLpro), one of two cysteine proteases responsible for the maturation and infectivity of SARS-CoV-2, processes and liberates functional proteins from the viral polyproteins and cleaves ubiquitin and ISG15 modifications to inhibit innate immune sensing. Consequently, PLpro is an attractive target for developing COVID-19 therapies. PLpro contains a zinc-finger domain important for substrate binding and structural stability. However, the impact of metal ions on the activity and biophysical properties of SARS-CoV-2 PLpro has not been comprehensively studied. Here, we assessed the impacts of metal ions on the catalytic activity of PLpro. Zinc had the largest inhibitory effect on PLpro, followed by manganese. Calcium, magnesium, and iron had smaller or no effects on PLpro activity. EDTA at a concentration of 0.5 mM was essential for PLpro activity, likely by chelating trace metals that inhibit PLpro. IC50 values for ZnCl2, ZnSO4, and MnCl2 of 0.42 ± 0.02 mM, 0.35 ± 0.01 mM, and 2.6 ± 0.3 mM were obtained in the presence of 0.5 mM EDTA; in the absence of EDTA, the estimated IC50 of ZnCl2 was 14 µM. Tryptophan intrinsic fluorescence analysis confirmed the binding of zinc and manganese to PLpro, and differential scanning calorimetry revealed that zinc but not manganese reduced ΔHcal of PLpro. The results of this study provide a reference for further work targeting PLpro to prevent and treat COVID-19.


Subject(s)
COVID-19 Drug Treatment , SARS-CoV-2 , Humans , Papain/chemistry , Papain/metabolism , Peptide Hydrolases/metabolism , Magnesium , Calcium , Tryptophan , Edetic Acid , Ubiquitin/metabolism , Polyproteins , Ions , Zinc , Iron
10.
Sci Rep ; 12(1): 16481, 2022 10 01.
Article in English | MEDLINE | ID: covidwho-2050552

ABSTRACT

Observations of air pollution in Krakow have shown that air quality has been improved during the last decade. In the presented study two factors affecting the physicochemical characteristic of PM2.5 fraction at AGH station in Krakow were observed. One is the ban of using solid fuels for heating purposes and the second is COVID-19 pandemic in Krakow. The PM2.5 fraction was collected during the whole year every 3rd day between 2nd March 2020 and 28th February 2021 at AGH station in Krakow. In total 110 PM2.5 fraction samples were collected. The chemical composition was determined for these samples. The elemental analysis was performed by energy dispersive X-ray fluorescence (EDXRF) technique, ions analysis was performed by ion chromatography (IC) and black carbon by optical method. In order to identify the emission sources the positive matrix factorization (PMF) was used. The results of such study were compared to similar analysis performed for PM2.5 for the period from June 2018 to May 2019 at AGH station in Krakow. The PM2.5 concentration dropped by 25% in 2020/2021 in comparison to 2018/2019 at this station. The concentrations of Si, K, Fe, Zn and Pb were lowering by 43-64% in the year 2020/2021 in comparison to 2018/2019. Cu, Mn, Zn and Pb come from mechanical abrasion of brakes and tires while Ti, Fe, Mn and Si are crustal species. They are the indicators of road dust (non-exhaust traffic source). Moreover, the annual average contribution of traffic/industrial/soil/construction work source was reduced in 2020/2021 in comparison to 2018/2019. As well the annual average contribution of fuels combustion was declining by 22% in 2020/2021 in comparison to 2018/2019. This study shows that the ban and lockdown, during COVID-19 pandemic, had significant impact on the characteristic of air pollution in Krakow.


Subject(s)
Air Pollutants , COVID-19 , Air Pollutants/analysis , COVID-19/epidemiology , Carbon/analysis , Communicable Disease Control , Dust/analysis , Environmental Monitoring/methods , Humans , Ions/analysis , Lead/analysis , Pandemics , Particulate Matter/analysis , Poland/epidemiology , Soil , Vehicle Emissions/analysis
11.
Inorg Chem ; 61(39): 15664-15677, 2022 Oct 03.
Article in English | MEDLINE | ID: covidwho-2036737

ABSTRACT

The identification of novel therapeutics against the pandemic SARS-CoV-2 infection is an indispensable new address of current scientific research. In the search for anti-SARS-CoV-2 agents as alternatives to the vaccine or immune therapeutics whose efficacy naturally degrades with the occurrence of new variants, the salts of Bi3+ have been found to decrease the activity of the Zn2+-dependent non-structural protein 13 (nsp13) helicase, a key component of the SARS-CoV-2 molecular tool kit. Here, we present a multilevel computational investigation based on the articulation of DFT calculations, classical MD simulations, and MIF analyses, focused on the examination of the effects of Bi3+/Zn2+ exchange on the structure and molecular interaction features of the nsp13 protein. Our calculations confirmed that Bi3+ ions can replace Zn2+ in the zinc-finger metal centers and cause slight but appreciable structural modifications in the zinc-binding domain of nsp13. Nevertheless, by employing an in-house-developed ATOMIF tool, we evidenced that such a Bi3+/Zn2+ exchange may decrease the extension of a specific hydrophobic portion of nsp13, responsible for the interaction with the nsp12 protein. The present study provides for a detailed, atomistic insight into the potential anti-SARS-CoV-2 activity of Bi3+ and, more generally, evidences the hampering of the nsp13-nsp12 interaction as a plausible therapeutic strategy.


Subject(s)
COVID-19 , SARS-CoV-2 , Bismuth , Humans , Ions , RNA Helicases/chemistry , RNA Helicases/metabolism , Salts , Zinc
12.
Rapid Commun Mass Spectrom ; 36(20): e9373, 2022 Oct 30.
Article in English | MEDLINE | ID: covidwho-2027400

ABSTRACT

RATIONALE: The COVID-19 pandemic demonstrated the importance of high-throughput analysis for public health. Given the importance of surface viral proteins for interactions with healthy tissue, they are targets of interest for mass spectrometry-based analysis. For that reason, the possibility of detecting and quantifying peptides using a high-throughput technique, laser diode thermal desorption-triple quadrupole mass spectrometry (LDTD-QqQMS), was explored. METHODS: Two peptides used as models for small peptides (leu-enkephalin and endomorphin-2) and four tryptic peptides (GVYYPDK, NIDGYFK, IADYNYK, and QIAPGQTGK) specific to the SARS-CoV-2 Spike protein were employed. Target peptides were analyzed individually in the positive mode by LDTD-QqQMS. Peptides were quantified by internal calibration using selected reaction monitoring transitions in pure solvents and in samples spiked with 20 µg mL-1 of a bovine serum albumin tryptic digest to represent real analysis conditions. RESULTS: Low-energy fragment ions (b and y ions) as well as high-energy fragment ions (c and x ions) and some of their corresponding water or ammonia losses were detected in the full mass spectra. Only for the smallest peptides, leu-enkephalin and endomorphin-2, were [M + H]+ ions observed. Product ion spectra confirmed that, with the experimental conditions used in the present study, LDTD transfers a considerable amount of energy to the target peptides. Quantitative analysis showed that it was possible to quantify peptides using LDTD-QqQMS with acceptable calibration curve linearity (R2 > 0.99), precision (RSD < 18.2%), and trueness (bias < 8.3%). CONCLUSIONS: This study demonstrated for the first time that linear peptides can be qualitatively and quantitatively analyzed using LDTD-QqQMS. Limits of quantification and dynamic ranges are still inadequate for clinical applications, but other applications where higher levels of proteins must be detected could be possible with LDTD. Given the high-throughput capabilities of LDTD-QqQMS (>15 000 samples in less than 43 h), more studies are needed to improve the sensitivity for peptide analysis of this technique.


Subject(s)
COVID-19 , Tandem Mass Spectrometry , Enkephalin, Leucine , Humans , Ions , Lasers , Pandemics , Peptides , SARS-CoV-2 , Spike Glycoprotein, Coronavirus , Tandem Mass Spectrometry/methods
13.
Int J Mol Sci ; 23(16)2022 Aug 12.
Article in English | MEDLINE | ID: covidwho-2023734

ABSTRACT

Heavy metal ions can disrupt biological functions via multiple molecular mechanisms, including inhibition of enzymes. We investigate the interactions of human papain-like cysteine endopeptidases cathepsins L, K, and S with gallium and cerium ions, which are associated with medical applications. We compare these results with zinc and lead, which are known to inhibit thiol enzymes. We show that Ga3+, Ce3+, and Ce4+ ions inhibit all tested peptidases with inhibition constants in the low micromolar range (between 0.5 µM and 10 µM) which is comparable to Zn2+ ions, whereas inhibition constants of Pb2+ ions are one order of magnitude higher (30 µM to 150 µM). All tested ions are linear specific inhibitors of cathepsin L, but cathepsins K and S are inhibited by Ga3+, Ce3+, and Ce4+ ions via hyperbolic inhibition mechanisms. This indicates a mode of interaction different from that of Zn2+ and Pb2+ ions, which act as linear specific inhibitors of all peptidases. All ions also inhibit the degradation of insoluble elastin, which is a common target of these peptidases in various inflammatory diseases. Our results suggest that these ions and their compounds have the potential to be used as cysteine cathepsin inhibitors in vitro and possibly in vivo.


Subject(s)
Cerium , Gallium , Cathepsin K/metabolism , Cathepsins/metabolism , Cysteine , Cysteine Proteinase Inhibitors/metabolism , Cysteine Proteinase Inhibitors/pharmacology , Endopeptidases/metabolism , Humans , Ions , Kinetics , Lead
14.
Sci Rep ; 12(1): 3794, 2022 03 08.
Article in English | MEDLINE | ID: covidwho-2004784

ABSTRACT

SARS-CoV-2 virions enter the host cells by docking their spike glycoproteins to the membrane-bound Angiotensin Converting Enzyme 2. After intracellular assembly, the newly formed virions are released from the infected cells to propagate the infection, using the extra-cytoplasmic ACE2 docking mechanism. However, the molecular events underpinning SARS-CoV-2 transmission between host cells are not fully understood. Here, we report the findings of a scanning Helium-ion microscopy study performed on Vero E6 cells infected with mNeonGreen-expressing SARS-CoV-2. Our data reveal, with unprecedented resolution, the presence of: (1) long tunneling nanotubes that connect two or more host cells over submillimeter distances; (2) large scale multiple cell fusion events (syncytia); and (3) abundant extracellular vesicles of various sizes. Taken together, these ultrastructural features describe a novel intra-cytoplasmic connection among SARS-CoV-2 infected cells that may act as an alternative route of viral transmission, disengaged from the well-known extra-cytoplasmic ACE2 docking mechanism. Such route may explain the elusiveness of SARS-CoV-2 to survive from the immune surveillance of the infected host.


Subject(s)
Microscopy/methods , SARS-CoV-2/physiology , Virus Internalization , Angiotensin-Converting Enzyme 2/metabolism , Animals , COVID-19/transmission , COVID-19/virology , Chlorocebus aethiops , Cytoplasm/chemistry , Cytoplasm/ultrastructure , Cytoplasm/virology , Extracellular Vesicles/chemistry , Extracellular Vesicles/ultrastructure , Giant Cells/chemistry , Giant Cells/physiology , Helium/chemistry , Humans , Ions/chemistry , SARS-CoV-2/isolation & purification , Spike Glycoprotein, Coronavirus/chemistry , Spike Glycoprotein, Coronavirus/metabolism , Vero Cells
15.
Langmuir ; 38(30): 9186-9194, 2022 08 02.
Article in English | MEDLINE | ID: covidwho-1947187

ABSTRACT

The spike (S) protein of SARS-CoV-2 has been found to play a decisive role in the cell entry mechanism of the virus and has been the prime target of most vaccine development efforts. Although numerous vaccines are already in use and more than half of the world population has been fully vaccinated, the emergence of new variants of the virus poses a challenge to the existing vaccines. Hence, developing an effective drug therapy is a crucial step in ending the pandemic. Nanoparticles can play a crucial role as a drug or a drug carrier and help tackle the pandemic effectively. Here, we performed explicit all-atom molecular dynamics simulations to probe interactions between S protein and Montmorillonite (MMT) nano clay surface. We built two systems with different counterions (Na+ and Ca2+), namely Na-MMT and Ca-MMT, to investigate the effect of different ions on S protein-MMT interaction. Structural modification of S protein was observed in the presence of MMT surface, particularly the loss of helical content of S protein. We revealed that electrostatic and hydrophobic interactions synergistically govern the S protein-MMT interactions. However, hydrophobic interactions were more pronounced in the Na-MMT system than in Ca-MMT. We also revealed residues and glycans of S protein closely interacting with the MMT surface. Interestingly, N165 and N343, which we found to be closely interacting with MMT in our simulations, also have a critical role in cell entry and in thwarting the cell's immune response in recent studies. Overall, our work provides atomistic insights into S protein-MMT interaction and enriches our understanding of the nanoparticle-S protein interaction mechanism, which will help develop advanced therapeutic techniques in the future.


Subject(s)
COVID-19 , Spike Glycoprotein, Coronavirus , Bentonite/chemistry , Humans , Ions , SARS-CoV-2 , Spike Glycoprotein, Coronavirus/metabolism
16.
Int J Mol Sci ; 21(16)2020 Aug 06.
Article in English | MEDLINE | ID: covidwho-1934101

ABSTRACT

The recently discovered 340-cavity in influenza neuraminidase (NA) N6 and N7 subtypes has introduced new possibilities for rational structure-based drug design. However, the plasticity of the 340-loop (residues 342-347) and the role of the 340-loop in NA activity and substrate binding have not been deeply exploited. Here, we investigate the mechanism of 340-cavity formation and demonstrate for the first time that seven of nine NA subtypes are able to adopt an open 340-cavity over 1.8 µs total molecular dynamics simulation time. The finding that the 340-loop plays a role in the sialic acid binding pathway suggests that the 340-cavity can function as a druggable pocket. Comparing the open and closed conformations of the 340-loop, the side chain orientation of residue 344 was found to govern the formation of the 340-cavity. Additionally, the conserved calcium ion was found to substantially influence the stability of the 340-loop. Our study provides dynamical evidence supporting the 340-cavity as a druggable hotspot at the atomic level and offers new structural insight in designing antiviral drugs.


Subject(s)
Antiviral Agents/pharmacology , Drug Development , Neuraminidase/chemistry , Orthomyxoviridae/enzymology , Binding Sites , Calcium/chemistry , Ions , Models, Molecular , Molecular Dynamics Simulation , N-Acetylneuraminic Acid/chemistry , Principal Component Analysis , Protein Structure, Secondary , Thermodynamics
17.
Cells ; 11(13)2022 06 23.
Article in English | MEDLINE | ID: covidwho-1933986

ABSTRACT

Two pore channels (TPCs) are implicated in vesicle trafficking, virus infection, and autophagy regulation. As Na+- or Ca2+-permeable channels, TPCs have been reported to be activated by NAADP, PI(3,5)P2, and/or high voltage. However, a comparative study on the function and regulation of the three mammalian TPC subtypes is currently lacking. Here, we used the electrophysiological recording of enlarged endolysosome vacuoles, inside-out and outside-out membrane patches to examine the three TPCs of rabbit (Oryctolagus cuniculus, or Oc) heterologously expressed in HEK293 cells. While PI(3,5)P2 evoked Na+ currents with a potency order of OcTPC1 > OcTPC3 > OcTPC2, only OcTPC2 displayed a strict dependence on PI(3,5)P2. Both OcTPC1 and OcTPC3 were activatable by PI3P and OcTPC3 was also activated by additional phosphoinositide species. While OcTPC2 was voltage-independent, OcTPC1 and OcTPC3 showed voltage dependence with OcTPC3 depending on high positive voltages. Finally, while OcTPC2 preferred a luminal pH of 4.6-6.0 in endolysosomes, OcTPC1 was strongly inhibited by extracytosolic pH 5.0 in both voltage-dependent and -independent manners, and OcTPC3 was inhibited by pH 6.0 but potentiated by pH 8.0. Thus, the three OcTPCs form phosphoinositide-activated Na+ channels with different ligand selectivity, voltage dependence, and extracytosolic pH sensitivity, which likely are optimally tuned for function in specific endolysosomal populations.


Subject(s)
Lysosomes , Phosphatidylinositols , Animals , HEK293 Cells , Humans , Hydrogen-Ion Concentration , Ions , Mammals , Phosphatidylinositol Phosphates , Rabbits
18.
World J Microbiol Biotechnol ; 38(9): 158, 2022 Jul 12.
Article in English | MEDLINE | ID: covidwho-1930505

ABSTRACT

In this mini-review, after a brief introduction into the widespread antimicrobial use of silver ions and nanoparticles against bacteria, fungi and viruses, the toxicity of silver compounds and the molecular mechanisms of microbial silver resistance are discussed, including recent studies on bacteria and fungi. The similarities and differences between silver ions and silver nanoparticles as antimicrobial agents are also mentioned. Regarding bacterial ionic silver resistance, the roles of the sil operon, silver cation efflux proteins, and copper-silver efflux systems are explained. The importance of bacterially produced exopolysaccharides as a physiological (biofilm) defense mechanism against silver nanoparticles is also emphasized. Regarding fungal silver resistance, the roles of metallothioneins, copper-transporting P-type ATPases and cell wall are discussed. Recent evolutionary engineering (adaptive laboratory evolution) studies are also discussed which revealed that silver resistance can evolve rapidly in bacteria and fungi. The cross-resistance observed between silver resistance and resistance to other heavy metals and antibiotics in bacteria and fungi is also explained as a clinically and environmentally important issue. The use of silver against bacterial and fungal biofilm formation is also discussed. Finally, the antiviral effects of silver and the use of silver nanoparticles against SARS-CoV-2 and other viruses are mentioned. To conclude, silver compounds are becoming increasingly important as antimicrobial agents, and their widespread use necessitates detailed understanding of microbial silver response and resistance mechanisms, as well as the ecological effects of silver compounds. Figure created with BioRender.com.


Subject(s)
Anti-Infective Agents , Bacterial Infections , COVID-19 , Metal Nanoparticles , Anti-Bacterial Agents/metabolism , Anti-Bacterial Agents/pharmacology , Anti-Infective Agents/metabolism , Anti-Infective Agents/pharmacology , Bacteria/metabolism , Copper/metabolism , Humans , Ions/metabolism , Ions/pharmacology , SARS-CoV-2 , Silver/metabolism , Silver/pharmacology , Silver Compounds/metabolism , Silver Compounds/pharmacology
19.
Bull Environ Contam Toxicol ; 108(5): 819-823, 2022 May.
Article in English | MEDLINE | ID: covidwho-1919758

ABSTRACT

Fine particulate matter (named PM2.5) has become a prominent and dangerous form of air pollution. The chemical composition of PM2.5 mainly includes inorganic elements, water soluble ions, elemental carbon (EC), organic carbon (OC), and organic compounds. The detection method for inorganic elements mainly includes X ray fluorescence, inductively coupled plasma-atomic emission spectrometry, and inductively coupled plasma mass spectrometry. As for water soluble ions, ion chromatography is the most common detection method. EC and OC are usually detected by carbon analyzer. The organic compounds are determined by gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry. In this paper, the merits and drawbacks of each analytical methods for the determination of PM2.5 chemical composition are summarized. This review also includes our discussion on the improvement of the analytical accuracy for the determination of PM2.5 chemical composition owing to the development of reference materials.


Subject(s)
Air Pollutants , Aerosols/analysis , Air Pollutants/analysis , Carbon/analysis , China , Environmental Monitoring/methods , Ions/analysis , Organic Chemicals/analysis , Particulate Matter/analysis , Seasons , Water/chemistry
20.
J Trace Elem Med Biol ; 73: 127015, 2022 Sep.
Article in English | MEDLINE | ID: covidwho-1867436

ABSTRACT

OBJECTIVE: Severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2), a worldwide health problem, is the cause of 2019 coronavirus disease. This study aimed to compare the trace element (selenium and iron), electrolyte (calcium and sodium), and physical activity levels of COVID-19 patients before and after COVID-19 treatment. METHOD: This prospective study was conducted in patients diagnosed with COVID-19 (n = 15). Trace element (selenium and iron), electrolyte (calcium and sodium), and physical activity levels of the patients were compared before and after the treatment. RESULT: Most of patients had selenium deficiency (86.7 %), iron deficiency (73.3 %), calcium deficiency (66.7 %) and sodium deficiency (46.7 %) before COVID-19 treatment. The most important improvements were seen in iron deficiency (from 73.3 % to 26.7 %) and sodium deficiency (from 46.7 % to 13.3 %) after the treatment. Selenium, iron, calcium, and sodium levels of the patients were significantly higher after the treatment (p < 0.05). The patients had low physical activity before and after COVID-19 treatment. In addition, no statistically significant difference was found in the comparison of physical activity levels (p > 0.05). CONCLUSION: This study indicated that selenium, iron, calcium, and sodium levels and deficiencies might improve after treating patients with COVID-19. However, the results of this study showed that the physical activity levels of COVID-19 patients might remain stable and low throughout the treatment process.


Subject(s)
COVID-19 Drug Treatment , Selenium , Trace Elements , Calcium , Electrolytes , Exercise , Humans , Ions , Iron , Prospective Studies , SARS-CoV-2 , Selenium/therapeutic use , Sodium , Trace Elements/therapeutic use
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