Journal: Biology Methods & Protocols

Article Title: An array-based melt curve analysis method for the identification and classification of closely related pathogen strains

doi: 10.1093/biomethods/bpy005

Figure Lengend Snippet: HEV and HRV target sequences

Article Snippet: Target-HRV-F , ATCC: VR-1663D , Commercial strain , na.

Techniques: Amplification, Sequencing, Binding Assay

Journal: Journal of drug delivery science and technology

Article Title: MATRIX-BASED CONTROLLED RELEASE DELIVERY OF ACYCLOVIR FROM POLY-(ETHYLENE CO-VINYL ACETATE) RINGS

doi: 10.1016/j.jddst.2019.101391

Figure Lengend Snippet: In vitro infection assays were performed to analyze the suppressive capabilities of the experimental rings compared to normally infected Vero cells with no treatment. Vero cells were either mock-infected (data not shown) or infected with either (A) HSV-1 (17); (B) HSV-1 (KOS); (C) HSV-2 (MS); or (D) HSV-2 (G) in the presence of no treatment, EVA25 rings, 65% ACV (w:w) EVA25 rings, or 50 μg/mL ACV diluted in α-MEM. Titers are expressed as log (pfu/mL) for each infection/treatment combination. Experiments were performed in triplicate.

Article Snippet: The monolayer of cells was infected with 100 μL of either HSV-1 (KOS), HSV-1 (17), HSV-2 (MS), HSV-2 (G) (ATCC #VR-1493 #VR-1789, #VR-540, and #VR-734, respectively) in Dulbecco’s phosphate buffered saline (PBS; Corning CellGro) at an MOI of approximately 1, or mock-infected with PBS.

Techniques: In Vitro, Infection

Journal: medRxiv

Article Title: COVID-19 wastewater based epidemiology: long-term monitoring of 10 WWTP in France reveals the importance of the sampling context

doi: 10.1101/2021.05.06.21256751

Figure Lengend Snippet: N1 recovery yield determination on a negative sample (NEG) after addition of heat inactivated SARS-CoV-2 (POS) in known quantity.

Article Snippet: To this end, a SARS-CoV-2 negative sample from WWTP1 (thereafter named NEG) sampled on August, 17 2020 was spiked with a known quantity of heat-inactivated SARS-CoV-2 (ATCC VR-1986HK™, thereafter referred as POS).

Techniques:

Journal: medRxiv

Article Title: COVID-19 wastewater based epidemiology: long-term monitoring of 10 WWTP in France reveals the importance of the sampling context

doi: 10.1101/2021.05.06.21256751

Figure Lengend Snippet: Raw N1-SARS-CoV-2 quantification (SARS-CoV-2, black circles and black moving average), incidence rate of “Département” (light grey curve) and rain (light grey bars) along with date for the 10 studied WWTPs.

Article Snippet: To this end, a SARS-CoV-2 negative sample from WWTP1 (thereafter named NEG) sampled on August, 17 2020 was spiked with a known quantity of heat-inactivated SARS-CoV-2 (ATCC VR-1986HK™, thereafter referred as POS).

Techniques:

Journal: medRxiv

Article Title: COVID-19 wastewater based epidemiology: long-term monitoring of 10 WWTP in France reveals the importance of the sampling context

doi: 10.1101/2021.05.06.21256751

Figure Lengend Snippet: Raw N1-SARS-CoV-2 quantification and contextualized N1-SARS-CoV-2 load (SARS-CoV-2 UG/mL, black circles and black moving average and SARS-CoV-2 10 10 UG/ 100 000 PE/d, black diamonds and dark grey moving average), incidence rate of “Département” (light grey curve) and rain (light grey bars) along with date for the 10 studied WWTPs.

Article Snippet: To this end, a SARS-CoV-2 negative sample from WWTP1 (thereafter named NEG) sampled on August, 17 2020 was spiked with a known quantity of heat-inactivated SARS-CoV-2 (ATCC VR-1986HK™, thereafter referred as POS).

Techniques:

Journal: bioRxiv

Article Title: Enisamium is a small molecule inhibitor of the influenza A virus and SARS-CoV-2 RNA polymerases

doi: 10.1101/2020.04.21.053017

Figure Lengend Snippet: Inhibition of IAV replication and FluPol activity by enisamium. (A) Effect of FAV00B on the replication of influenza A/WSN/33 (H1N1) virus. Data points represent mean and standard deviation of n=3 independent enisamium titrations and matching virus plaque experiments. (B) Effect of FAV00B on the steady-state levels of IAV vRNA, 5S rRNA and the expression of GFP. Levels of 5S rRNA and IAV vRNA were analysed by primer extension (middle panel). PA and tubulin expression were analysed by western blot (bottom panel). A mutant IAV FluPol containing two amino acid substitutions in the PB1 active site (PB1a) was used as negative control. Data points represent mean and standard deviation of n=3 independent enisamium titrations and matching GFP measurements or primer extensions. (C) Effect of T-705 triphosphate, FAV00A and VR17-04 on FluPol activity in vitro . Transcription products synthesised by FluPol in the presence of VR17-04 were analysed by PAGE (lower panel). Data points represent mean and standard deviation of n=3 independent enisamium titrations in RNA polymerase assays. (D) Effect of extracts from A549 cells treated with FAV00A on FluPol activity in vitro . RNA polymerase products were analysed by PAGE.

Article Snippet: Furthermore, the IC 50 value of VR17-04 against nsp7/8/12 is similar to that of remdesivir triphosphate ( ).

Techniques: Inhibition, Activity Assay, Standard Deviation, Expressing, Western Blot, Mutagenesis, Negative Control, In Vitro

Journal: bioRxiv

Article Title: Enisamium is a small molecule inhibitor of the influenza A virus and SARS-CoV-2 RNA polymerases

doi: 10.1101/2020.04.21.053017

Figure Lengend Snippet: List of compounds tested and their IC 50 values in FluPol in vitro RNA synthesis assays.

Article Snippet: Furthermore, the IC 50 value of VR17-04 against nsp7/8/12 is similar to that of remdesivir triphosphate ( ).

Techniques: In Vitro

Journal: bioRxiv

Article Title: Enisamium is a small molecule inhibitor of the influenza A virus and SARS-CoV-2 RNA polymerases

doi: 10.1101/2020.04.21.053017

Figure Lengend Snippet: Inhibition of FluPol transcription by VR17-04. (A) Schematic of FluPol transcription initiation using a capped RNA primer. The terminal AG bases of the primer can base pair with the terminal U (orange), or the terminal UC (red) of the influenza virus template, resulting in two possible extension products called the C- and G-product, respectively. (B) Effect of VR17-04 on FluPol transcription in vitro using a radiolabelled capped RNA as primer. An active site mutant containing a mutation in the PB1 active site was used as negative control. DMSO was used a solvent control. C- and G-products are indicated as well as realignment products that can form when the polymerase slips at position 4 of the template.

Article Snippet: Furthermore, the IC 50 value of VR17-04 against nsp7/8/12 is similar to that of remdesivir triphosphate ( ).

Techniques: Inhibition, In Vitro, Mutagenesis, Negative Control

Journal: bioRxiv

Article Title: Enisamium is a small molecule inhibitor of the influenza A virus and SARS-CoV-2 RNA polymerases

doi: 10.1101/2020.04.21.053017

Figure Lengend Snippet: Purification and RNA synthesis activity of SARS-CoV-2 nsp7/8/12 complex. (A) SARS-CoV-2 nsp7, nsp8 and nsp12 were expressed and purified from E. coli (nsp7 and nsp8) or Sf9 (nsp12) cells and visualised by SDS PAGE. (B) Schematic of 40 nt RNA template and 20 nt radiolabelled primer. (C) Purified nsp7/8/12 complex was tested for in vitro activity in the presence or absence of NTPs and RNA template. The 20 nt primer and 40 nt product ran slightly slower than the markers, possibly due to differences in the RNA sequence or phosphorylation state. Quantification of the transcription products is shown on the right. Quantification is from n=3 independently prepared reactions using the same nsp7/8/12 protein preparation, error bars represent standard deviation.

Article Snippet: Furthermore, the IC 50 value of VR17-04 against nsp7/8/12 is similar to that of remdesivir triphosphate ( ).

Techniques: Purification, Activity Assay, SDS Page, In Vitro, Sequencing, Standard Deviation

Journal: bioRxiv

Article Title: Enisamium is a small molecule inhibitor of the influenza A virus and SARS-CoV-2 RNA polymerases

doi: 10.1101/2020.04.21.053017

Figure Lengend Snippet: Inhibition of SARS-CoV-2 nsp7/8/12 by remdesivir triphosphate, T-705 triphosphate, enisamium and VR17-04. (A) Effect of remdesivir triphosphate and T-705 triphosphate on nsp7/8/12 activity in vitro . Polymerase activity was plotted against drug concentration and dose-response curves were fit to the data (right). (B) Effect of FAV00A and VR17-04 on nsp7/8/12 activity in vitro . Polymerase activity was plotted against drug concentration and dose-response curves were fit to the data (right). Quantification is from n=3-6 independently prepared reactions using the same nsp7/8/12 protein preparation, error bars represent standard deviation. (C) IC 50 values of remdesivir triphosphate, T-705 triphosphate, FAV00A and VR17-04 for IAV FluPol and SARS-CoV-2 nsp7/8/12. The IC 50 value for VR17-04 on SARS-CoV-2 nsp7/8/12 could not be calculated and therefore it was estimated from the dose-response curve.

Article Snippet: Furthermore, the IC 50 value of VR17-04 against nsp7/8/12 is similar to that of remdesivir triphosphate ( ).

Techniques: Inhibition, Activity Assay, In Vitro, Concentration Assay, Standard Deviation