scholarly journals The Effect of Allicin on the Proteome of SARS-CoV-2 Infected Calu-3 Cells

2021 ◽  
Vol 12 ◽  
Author(s):  
Kirstin Mösbauer ◽  
Verena Nadin Fritsch ◽  
Lorenz Adrian ◽  
Jörg Bernhardt ◽  
Martin Clemens Horst Gruhlke ◽  
...  
Keyword(s):  
Organosulfur Compound ◽  
Toxicity Tests ◽  
Immunomodulatory Activity ◽  
Label Free ◽  
Lead Compounds ◽  
Viral Particles ◽  
Protein Thiols ◽  
Thiol Reactivity ◽  
Future Drug ◽  
Bacteria And Fungi

Allicin (diallyl thiosulfinate) is the major thiol-reactive organosulfur compound produced by garlic plants (Allium sativum) upon tissue damage. Allicin exerts its strong antimicrobial activity against bacteria and fungi via S-thioallylation of protein thiols and low molecular weight thiols. Here, we investigated the effect of allicin on SARS-CoV-2 infected Vero E6 and Calu-3 cells. Toxicity tests revealed that Calu-3 cells showed greater allicin tolerance, probably due to >4-fold higher GSH levels compared to the very sensitive Vero E6 cells. Exposure of infected Vero E6 and Calu-3 cells to biocompatible allicin doses led to a ∼60–70% decrease of viral RNA and infectious viral particles. Label-free quantitative proteomics was used to investigate the changes in the Calu-3 proteome after SARS-CoV-2 infection and the effect of allicin on the host-virus proteome. SARS-CoV-2 infection of Calu-3 cells caused a strong induction of the antiviral interferon-stimulated gene (ISG) signature, including several antiviral effectors, such as cGAS, Mx1, IFIT, IFIH, IFI16, IFI44, OAS, and ISG15, pathways of vesicular transport, tight junctions (KIF5A/B/C, OSBPL2, CLTCL1, and ARHGAP17) and ubiquitin modification (UBE2L3/5), as well as reprogramming of host metabolism, transcription and translation. Allicin treatment of infected Calu-3 cells reduced the expression of IFN signaling pathways and ISG effectors and reverted several host pathways to levels of uninfected cells. Allicin further reduced the abundance of the structural viral proteins N, M, S and ORF3 in the host-virus proteome. In conclusion, our data demonstrate the antiviral and immunomodulatory activity of biocompatible doses of allicin in SARS-CoV-2-infected cell cultures. Future drug research should be directed to exploit the thiol-reactivity of allicin derivatives with increased stability and lower human cell toxicity as antiviral lead compounds.

scholarly journals Cellular Dielectric Spectroscopy: A Powerful New Approach to Label-Free Cellular Analysis

2004 ◽  
Vol 9 (6) ◽  
pp. 467-480 ◽  
Author(s):  
Gary J. Ciambrone ◽  
Vivian F. Liu ◽  
Deborah C. Lin ◽  
Ryan P. McGuinness ◽  
Gordon K. Leung ◽  
...  
Keyword(s):  
Signal Transduction ◽  
Dielectric Spectroscopy ◽  
Protein Tyrosine Kinase ◽  
Label Free ◽  
Biologically Relevant ◽  
Lead Compounds ◽  
Cellular Analysis ◽  
Cellular Signal ◽  
Receptor Signaling Pathway ◽  
Specific Frequency

The past decade has seen a number of significant changes in identifying higher quality lead compounds earlier in the drug discovery process. Cell-based assay technologies yielding high-content information have emerged to achieve this goal. Although most of these systems are based on fluorescence detection, this article describes the development and application of an innovative cellular assay technology based on radio frequency spectrometry and bioimpedance measurements. Using this technique, the authors have discovered a link between cellular bioimpedance changes and receptor-mediated signal transduction events. By performing dielectric spectroscopy of cells across as pectrum of frequencies (1 KHz to 110 MHz), a series of receptor-specific, frequency-dependent impedance patterns is collected. These raw data patterns are used to determine the identity of the cellular receptor-signaling pathway being tested and to quantify stimulation endpoints and kinetics. The authors describe the application of this technology to the analysis of ligand-induced cellular responses mediated by the 3 major classes of G-protein-coupled receptors (GPCRs) and protein tyrosine kinase receptors. This single assay platform can be used with ease to monitor Gs, Gi, and Gq GPCRs without the need for chimeric or promiscuous G-proteins, fluorophors, or tagged proteins. In contrast to other methods of monitoring cellular signal transduction, this approach provides high information content in a simplified, noninvasive, and biologically relevant fashion.

scholarly journals Natural Products as Potential Lead Compounds for Drug Discovery Against SARS-CoV-2

2021 ◽  
Author(s):  
Oyere Tanyi Ebob ◽  
Smith B. Babiaka ◽  
Fidele Ntie-Kang
Keyword(s):  
Natural Products ◽  
Drug Discovery ◽  
Infectious Diseases ◽  
Review Article ◽  
Literature Survey ◽  
Lead Compounds ◽  
The Past ◽  
The World ◽  
Bacteria And Fungi ◽  
New Vaccines

AbstractFor the past 2 years, the coronavirus responsible for the COVID-19 infection has become a world pandemic, ruining the lives and economies of several nations in the world. This has scaled up research on the virus and the resulting infection with the goal of developing new vaccines and therapies. Natural products are known to be a rich source of lead compounds for drug discovery, including against infectious diseases caused by microbes (viruses, bacteria and fungi). In this review article, we conducted a literature survey aimed at identifying natural products with inhibitory concentrations against the coronaviruses or their target proteins, which lie below 10 µM. This led to the identification of 42 compounds belonging to the alkaloid, flavonoid, terpenoid, phenolic, xanthone and saponin classes. The cut off concentration of 10 µM was to limit the study to the most potent chemical entities, which could be developed into therapies against the viral infection to make a contribution towards limiting the spread of the disease.

The Genus Glycosmis [Rutaceae]: A Comprehensive Review on its Phytochemical and Pharmacological Perspectives

2019 ◽  
Vol 9 (2) ◽  
pp. 98-124 ◽  
Author(s):  
Mohammad Yasir ◽  
Manish K. Tripathi ◽  
Pushpendra Singh ◽  
Rahul Shrivastava
Keyword(s):  
Complete Information ◽  
Convincing Evidence ◽  
Rich Source ◽  
Biologically Active ◽  
Immunomodulatory Activity ◽  
Anticancer Activities ◽  
Pharmacological Activities ◽  
Lead Compounds ◽  
The Family ◽  
Different Parts

Background:Medicinal plants are rich source of large variety of lead compounds. The plants of genus Glycosmis or its different parts valued for its therapeutic and medicinal qualities. Glycosmis belongs to the family Rutaceae.Objective:The genus Glycosmis is a rich source of pharmacologically and biologically active secondary metabolites, such as alkaloids, flavonoids, phenolic glycosides, quinones, terpenoids, glycerides isolated from different parts of Glycosmis plant. The objectives of this review is to provide updatedand complete information on the distribution, phytochemical, pharmacological, and toxicity research of Glycosmis species.Results:About 233 phytoconstituents are isolated from this genus. Recent interest in this genus has focused on isolating and identifying of different phytoconstituents that exhibit potent antioxidant, antibacterial, antiviral and anticancer activities. They show good cytotoxic activity against various cancer cell lines and also reported for good antiviral and immunomodulatory activity. In this support convincing evidence in experimental animal models are available.Conclusion:This review summarizes information about the isolated compounds their bioactivities related to same compounds present in other plants including pharmacological activities of plant extract of Glycosmis genus.

scholarly journals Label-free SARS-CoV-2 detection and classification using phase imaging with computational specificity

2021 ◽  
Vol 10 (1) ◽  
Author(s):  
Neha Goswami ◽  
Yuchen R. He ◽  
Yu-Heng Deng ◽  
Chamteut Oh ◽  
Nahil Sobh ◽  
...  
Keyword(s):  
Neural Network ◽  
Influenza A ◽  
Digital Pathology ◽  
Ground Truth ◽  
Optical Path Length ◽  
Phase Imaging ◽  
Processing Unit ◽  
Label Free ◽  
Interferometric Method ◽  
Viral Particles

AbstractEfforts to mitigate the COVID-19 crisis revealed that fast, accurate, and scalable testing is crucial for curbing the current impact and that of future pandemics. We propose an optical method for directly imaging unlabeled viral particles and using deep learning for detection and classification. An ultrasensitive interferometric method was used to image four virus types with nanoscale optical path-length sensitivity. Pairing these data with fluorescence images for ground truth, we trained semantic segmentation models based on U-Net, a particular type of convolutional neural network. The trained network was applied to classify the viruses from the interferometric images only, containing simultaneously SARS-CoV-2, H1N1 (influenza-A virus), HAdV (adenovirus), and ZIKV (Zika virus). Remarkably, due to the nanoscale sensitivity in the input data, the neural network was able to identify SARS-CoV-2 vs. the other viruses with 96% accuracy. The inference time for each image is 60 ms, on a common graphic-processing unit. This approach of directly imaging unlabeled viral particles may provide an extremely fast test, of less than a minute per patient. As the imaging instrument operates on regular glass slides, we envision this method as potentially testing on patient breath condensates. The necessary high throughput can be achieved by translating concepts from digital pathology, where a microscope can scan hundreds of slides automatically.

Carbohydrazones of Substituted Salicylaldehydes as Potential Lead Compounds for the Development of Narrow-Spectrum Antimicrobials

2007 ◽  
Vol 62 (7-8) ◽  
pp. 483-486 ◽  
Author(s):  
Eila Pelttari ◽  
Eliisa Karhumäki ◽  
Jane Langshaw ◽  
Hannu Elo
Keyword(s):  
Antimicrobial Properties ◽  
Test Organism ◽  
Diffusion Method ◽  
Antimicrobial Drugs ◽  
Antimicrobial Spectrum ◽  
Lead Compounds ◽  
Narrow Spectrum ◽  
Free Aldehyde ◽  
Bacteria And Fungi ◽  
Derivatives Of

Certain substituted salicylaldehydes are known to have highly potent antimicrobial activity against bacteria and fungi, but the mechanism underlying this remarkable activity is not known, and almost nothing has been reported on the effects of further modification of the structures, such as the formation of hydrazone-type derivatives. We report now a study on the antimicrobial properties of the carbohydrazone derivatives of several substituted salicylaldehydes. The compounds studied were synthesized from ring-substituted salicylaldehydes and carbohydrazide in the mole ratio 2:1. They were tested against Aspergillus niger, Bacillus cereus, Candida albicans, Escherichia coli, Pseudomonas aeruginosa, Saccharomyces cerevisiae and Staphylococcus epidermidis using the agar diffusion method. The carbohydrazone derived from 2,3,4-trihydroxybenzaldehyde had distinctly higher activity than the parent aldehyde in the same molar concentration. This activity was limited to one test organism (S. epidermidis), while the free aldehyde had at least some (in some cases even high) activity against all of the microbes studied. All other ones of the effective carbohydrazone compounds were distinctly less active than the parent salicylaldehydes as such. The hydrazones studied had in general a narrower antimicrobial spectrum than the free aldehydes and are thus of interest as potential lead compounds for the development of narrow-spectrum antimicrobial drugs. The mechanism of action of the aldehydes as well as that of the carbohydrazones is discussed

Label-Free Protein and Pathogen Detection Using the Atomic Force Microscope

2004 ◽  
Vol 9 (6) ◽  
pp. 491-497 ◽  
Author(s):  
Janice L. Huff ◽  
Michael P. Lynch ◽  
Saju Nettikadan ◽  
James C. Johnson ◽  
Srikanth Vengasandra ◽  
...  
Keyword(s):  
Biological Sciences ◽  
Atomic Force Microscope ◽  
Pathogen Detection ◽  
Label Free ◽  
Time Data ◽  
Surface Profiling ◽  
Atomic Force ◽  
Viral Particles ◽  
Label Free Detection ◽  
Direct Label

The atomic force microscope (AFM) uses a sharp micron-scale tip to scan and amplify surface features, providing exceptionally detailed topographical information with magnification on the order of ×106. This instrument is used extensively for quality control in the computer and semiconductor industries and is becoming a progressively more important tool in the biological sciences. Advantages of the AFM for biological application include the ability to obtain information in a direct, label-free manner and the ability to image in solution, providing real-time data acquisition under physiologically relevant conditions. A novel application of the AFM currently under development combines its surface profiling capabilities with fixed immuno-capture using antibodies immobilized in a nanoarray format. This provides a distinctive platform for direct, label-free detection and characterization of viral particles and other pathogens.

scholarly journals Biological Characteristics of “Wartowice” Post-Flotation Tailings Pond (Lower Silesia, Poland)

2014 ◽  
Vol 40 (1) ◽  
pp. 71-86 ◽  
Author(s):  
Justyna Rybak ◽  
Barbara Kołwzan ◽  
Kazimierz Grabas ◽  
Grzegorz Pasternak ◽  
Małgorzata Krawczyńska
Keyword(s):  
Metal Tolerance ◽  
Soil Microflora ◽  
Toxicity Tests ◽  
Dispersal Capacity ◽  
Tailings Pond ◽  
Toxicological Studies ◽  
Bacteria And Fungi ◽  
Microbiological Analyses ◽  
Number Of Bacteria ◽  
Lower Silesia

Abstract “Wartowice” tailings pond was closed in 1989, resulting in 232,4 ha tailings pile requiring reclamation. The major problem is heavy metals presence and poor nutrient conditions and physicochemical structure of soil which disturbs the plants development. In order to assess the real condition of studied area the complete biological characteristic has been done. The physicochemical conditions were assessed altogether with phytosociological, microbiological and toxicological studies of deposits. We recorded only 27 species of vascular plants belonging to 15 families on the tailings pond of which 5 belong to Rosaceae, 4 to Asteraceae and 3 to Poaceae and Saliceae. Species inhabiting the tailings depended on their dispersal capacity, metal tolerance and rhizome strategy. Microbiological analyses revealed the low number of bacteria and fungi on the tailings pond, apart from the small uplift area where the plants were indentified. Bacteria identified on the tailings pond were classified to 8 genera. The low number of bacteria suggests the lack of nutrients which affects the development of soil microflora. Toxicity tests showed that post-flotation sludge is not toxic to microorganisms because of its high pH. Some plants, such as lucerne could even influence positively the microorganisms development what has been proved in our studies. The tailings toxicity was higher towards producers, where Secale cereale appeared to be the most sensitive species. Amendment with topsoil from adjacent areas can influence positively the phytotoxic properties of tailings and enrich them into native seeds.

Electron Microscopy of a Herpesvirus Isolated from Marek's Disease in Duck and Chicken Embryo Fibroblast Cultures

1968 ◽  
Vol 26 ◽  
pp. 222-223 ◽  
Author(s):  
Keyvan Nazerian
Keyword(s):  
Inclusion Bodies ◽  
Chicken Embryo Fibroblast ◽  
Marek's Disease ◽  
Marek’S Disease ◽  
Duck Embryo Fibroblast ◽  
Multinucleated Cells ◽  
Viral Particles ◽  
Infected Cells ◽  
And Control ◽  
Do So

A herpes-like virus has been isolated from duck embryo fibroblast (DEF) cultures inoculated with blood from Marek's disease (MD) infected birds. Cultures which contained this virus produced MD in susceptible chickens while virus negative cultures and control cultures failed to do so. This and other circumstantial evidence including similarities in properties of the virus and the MD agent implicate this virus in the etiology of MD.Histochemical studies demonstrated the presence of DNA-staining intranuclear inclusion bodies in polykarocytes in infected cultures. Distinct nucleo-plasmic aggregates were also seen in sections of similar multinucleated cells examined with the electron microscope. These aggregates are probably the same as the inclusion bodies seen with the light microscope. Naked viral particles were observed in the nucleus of infected cells within or on the edges of the nucleoplasmic aggregates. These particles measured 95-100mμ, in diameter and rarely escaped into the cytoplasm or nuclear vesicles by budding through the nuclear membrane (Fig. 1). The enveloped particles (Fig. 2) formed in this manner measured 150-170mμ in diameter and always had a densely stained nucleoid. The virus in supernatant fluids consisted of naked capsids with 162 hollow, cylindrical capsomeres (Fig. 3). Enveloped particles were not seen in such preparations.

Detection of viruses by electron microscopy: Increased sensitivity by direct micro-ultracentrifugation of samples

1987 ◽  
Vol 45 ◽  
pp. 908-909
Author(s):  
Alain R. Trudel ◽  
M. Trudel
Keyword(s):  
Electron Microscopy ◽  
Fold Increase ◽  
Observation Time ◽  
Clinical Samples ◽  
Maximum Speed ◽  
Viral Particles ◽  
Structural Details ◽  
Latex Spheres ◽  
Increased Sensitivity ◽  
Size Of Particles

AirfugeR (Beckman) direct ultracentrifugation of viral samples on electron microscopy grids offers a rapid way to concentrate viral particles or subunits and facilitate their detection and study. Using the A-100 fixed angle rotor (30°) with a K factor of 19 at maximum speed (95 000 rpm), samples up to 240 μl can be prepared for electron microscopy observation in a few minutes: observation time is decreased and structural details are highlighted. Using latex spheres to calculate the increase in sensitivity compared to the inverted drop procedure, we obtained a 10 to 40 fold increase in sensitivity depending on the size of particles. This technique also permits quantification of viral particles in samples if an aliquot is mixed with latex spheres of known concentration.Direct ultracentrifugation for electron microscopy can be performed on laboratory samples such as gradient or column fractions, infected cell supernatant, or on clinical samples such as urine, tears, cephalo-rachidian liquid, etc..

A rapid method for the direct identification of paramyxovirus in canine CSF by ultracentrifugation and negative staining as a rule out for distemper

1990 ◽  
Vol 48 (3) ◽  
pp. 594-595
Author(s):  
W.L. Steffens ◽  
M.B. Ard ◽  
C.E. Greene ◽  
A. Jaggy
Keyword(s):  
Subacute Sclerosing Panencephalitis ◽  
Clinical Signs ◽  
Viral Disease ◽  
Etiologic Agent ◽  
Mucosal Inflammation ◽  
Worldwide Distribution ◽  
Negative Stain ◽  
Viral Particles ◽  
Systemic Manifestations ◽  
Rule Out

Canine distemper is a multisystemic contagious viral disease having a worldwide distribution, a high mortality rate, and significant central neurologic system (CNS) complications. In its systemic manifestations, it is often presumptively diagnosed on the basis of clinical signs and history. Few definitive antemortem diagnostic tests exist, and most are limited to the detection of viral antigen by immunofluorescence techniques on tissues or cytologic specimens or high immunoglobulin levels in CSF (cerebrospinal fluid). Diagnosis of CNS distemper is often unreliable due to the relatively low cell count in CSF (<50 cells/μl) and the binding of blocking immunoglobulins in CSF to cell surfaces. A more reliable and definitive test might be possible utilizing direct morphologic detection of the etiologic agent. Distemper is the canine equivalent of human measles, in that both involve a closely related member of the Paramyxoviridae, both produce mucosal inflammation, and may produce CNS complications. In humans, diagnosis of measles-induced subacute sclerosing panencephalitis is through negative stain identification of whole or incomplete viral particles in patient CSF.

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