Journal: mBio

Article Title: Hijacking the Fusion Complex of Human Parainfluenza Virus as an Antiviral Strategy

doi: 10.1128/mBio.03203-19

Figure Lengend Snippet: Dose-response of the new preactivating compound compared to that of the previously published prototype. (A) The HPIV3 clinical isolate was incubated with the indicated concentrations of compounds or left untreated for 60 min at 4°C or 37°C. The compounds were removed, and viral infectivity was determined by a plaque reduction assay. The effect of pretreatment with the compounds on infectivity is shown as the level of inhibition of the plaques as a percentage of the number inhibited without preincubation with compounds. (B) Chemical structures of PAC-3066 and CM9. (C) HPIV3 CI-1 eGFP was treated with different concentrations (5 mM, serially diluted 2-fold six times down to 0.078 mM) of the indicated small-molecule compounds for 1 h at 37°C. RNA was extracted from the remaining virus-compound solutions and used for viral genome quantification. The results shown are those obtained after treatment with 5 mM each compound. GCN, genome copy number.

Article Snippet: Using a one-step quantitative real time-PCR HPIV3 detection kit (Primer Design Ltd.) and Realplex2 Mastercycler (Eppendorf), the total amount of viral RNA in each sample was determined ( ).

Techniques: Incubation, Infection, Inhibition, Virus

Journal: mBio

Article Title: Hijacking the Fusion Complex of Human Parainfluenza Virus as an Antiviral Strategy

doi: 10.1128/mBio.03203-19

Figure Lengend Snippet: PAC-3066 does not affect HN-receptor interaction or neuraminidase activity. (A) Sensitivity of HN-receptor binding to viral entry inhibitors of cell monolayers expressing clinical isolate HPIV3 HN, incubated with either PAC-3066, zanamivir, or DMSO. Percent inhibition relative to that for DMSO-treated cells is shown. Each point represents the mean from 3 experiments, each of which was performed in triplicate, ± SEM. (B) The inhibition of neuraminidase activity in the presence or absence of the indicated compounds was assayed at 37°C and pH 5 on cell monolayers transiently expressing HPIV3 clinical isolate HN. Each bar represents the results from triplicate experiments, each of which was performed in triplicate, ± SEM.

Article Snippet: Using a one-step quantitative real time-PCR HPIV3 detection kit (Primer Design Ltd.) and Realplex2 Mastercycler (Eppendorf), the total amount of viral RNA in each sample was determined ( ).

Techniques: Activity Assay, Binding Assay, Expressing, Incubation, Inhibition

Journal: mBio

Article Title: Hijacking the Fusion Complex of Human Parainfluenza Virus as an Antiviral Strategy

doi: 10.1128/mBio.03203-19

Figure Lengend Snippet: Conformation change in F induced by PAC-3066 at 37°C visualized by cryo-electron tomography. (A, B) Overview of HPIV3 without (A) and with (B) PAC-3066. Bars, 50 nm. (C, D) Close-up of the viral surface glycoproteins without (C) and with (D) PAC-3066. (E) Subtomogram averages from a subset of viral surface glycoproteins where prefusion F can be identified without PAC-3066. (F, G) Structural comparison coordinates of HN (PDB accession number 4MZA ) and prefusion F (PDB accession number 6MJZ ) are shown in pink and cyan, respectively. (H) Subtomogram averages where postfusion F can be identified after PAC-3066 incubation. (I) Structural comparison coordinates of the postfusion F (PDB accession number 1ZTM ) are shown in light blue.

Article Snippet: Using a one-step quantitative real time-PCR HPIV3 detection kit (Primer Design Ltd.) and Realplex2 Mastercycler (Eppendorf), the total amount of viral RNA in each sample was determined ( ).

Techniques: Tomography, Comparison, Incubation

Journal: PLoS Neglected Tropical Diseases

Article Title: Susceptibility of Armigeres subalbatus Coquillett (Diptera: Culicidae) to Zika virus through oral and urine infection

doi: 10.1371/journal.pntd.0008450

Figure Lengend Snippet: ZIKV infection rate and quantification of Zika viral RNA copies, by real-time RT-PCR, detected in the salivary gland, midgut, ovary and collected saliva of Ar . subalbatus females at 4, 7, and 10 dpe (blood meal).

Article Snippet: ZIKV in these organs was detected using a One-Step Detection Kit for Zika Virus RNA (Daan Gene, Guangzhou, China), and PCR was conducted as previously described Li et al., 2017 [ ].

Techniques: Infection, Quantitative RT-PCR