Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: VeroE6 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir, narlaprevir, simeprevir, paritaprevir, grazoprevir, glecaprevir, voxilaprevir, vaniprevir, danoprevir, deldeprevir, asunaprevir and faldaprevir, as well as HCV NS4A inhibitor ACH-806, as described in Materials and Methods. After 46-50 hours of incubation, SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of counts from 7 replicate cultures ± standard errors of the means (SEM) and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay, as described in Materials and Methods. Datapoints (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance from 12 replicate nontreated control cultures. Sigmoidal concentration response curves were fitted and CC50 values were determined as shown in Supplementary Figure 3. The red / blue stippled line represents the drug concentrations at which DMSO is expected to induce antiviral effects with reduction of residual infectivity to <70% / cytotoxicity with reduction of cell viability to <90%, according to Supplementary Figure 2.

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: Huh7.5 cells were seeded in 96-well plates and the following day infected with SARS-CoV-2 followed by treatment with specified concentrations of the PI boceprevir, telaprevir and simeprevir, as described in Materials and Methods. After 70-74 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for the SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Datapoints (red dots) are means of 7 replicates ± SEM and represent % residual infectivity, determined as % SARS-CoV-2 positive cells relative to means of counts from 14 replicate infected nontreated control cultures. Sigmoidal concentration response curves (red lines) were fitted and EC50 values were determined, as described in Materials and Methods. Cell viability data were obtained in replicate assays with noninfected cells using a colorimetric assay as described in Materials and Methods. Data points (blue triangles) are means of 3 replicate cultures ± SEM and represent % cell viability relative to mean absorbance of 12 nontreated controls. Sigmoidal concentration response curves were fitted and CC50 values were determined, as shown in Supplementary Figure 4. The blue stippled line represents the drug concentrations at which DMSO is expected to induce cytotoxicity with reduction of cell viability to <90%, according to Supplementary Figure 2; DMSO did not induce antiviral effects in the tested dose ranges (Supplementary Figure 2).

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Incubation, Immunostaining, Concentration Assay, Colorimetric Assay

Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: VeroE6 cells seeded in 96-well plates were infected the following day with SARS-CoV-2 followed by treatment with specified concentrations of the linear PI boceprevir (BOC) and narlaprevir (NAR), or the macrocyclic PI simeprevir (SIM), paritaprevir (PAR) and grazoprevir (GRA), or polymerase inhibitor remdesivir (REM), or a combination of these PI and remdesivir, as described in Materials and Methods. After 46-50 hours incubation SARS-CoV-2 infected cells were visualized by immunostaining for SARS-CoV-2 Spike protein and quantified by automated counting, as described in Materials and Methods. Fractional effect (Fa) values were calculated by relating counts from infected and treated cultures to the mean count from at least 21 infected nontreated cultures and were entered into CompuSyn software. Datapoints are means of 6 to 7 replicates, and for each treatment experiment 6 to 10 datapoints were entered. For each inhibitor combination depicted per row, the following curves were fitted using Compusyn: (A) concentration-Fa curves plotting Fa values ranging from 0.01 to 0.99 against specified inhibitor concentrations. (B) Fa-CI curves plotting CI values ranging from 0 to 2 against Fa values ranging from 0.01 to 0.99. (C) Fa-Log 10 CI curves plotting logarithmic CI values ranging from 0.01 to 100 against Fa values ranging from 0.01 to 0.99. (B and C) Overall, CI values ≥1.1 suggest antagonism “A”, while CI values <0.9 suggest synergism “S” indicated by green and red colour coding of the y-axis, respectively. (D) Fa-DRI curves plotting DRI values ranging from 0 to 5 against Fa values ranging from 0.01 to 0.99; the datapoints and curve for simeprevir are outside the graph area and can be seen in (E). (E) Fa-Log 10 DRI curves plotting logarithmic DRI values ranging from 0.01 to 100 against Fa values ranging from 0.01 to 0.99. For an overview of CI and DRI values see .

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Incubation, Immunostaining, Software, Concentration Assay

Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: VeroE6 cells seeded the previous day in T25 flasks were infected with SARS-CoV-2 followed by treatment with 1-fold EC50 of PI boceprevir, telaprevir, narlaprevir, simeprevir, paritaprevir, grazoprevir, vaniprevir, danoprevir, asunaprevir and faldaprevir, which were administered immediately after infection and on day 1, 3, 5 and 7 post infection when cells were split, as described in Materials and Methods. Left panel, the % of SARS-CoV-2 infected cells on the specified days post infection, was determined by immunostaining. Right panel, replicate cultures were derived following cell splitting and treatment and immunostained for the SARS-CoV-2 Spike protein and counterstained with Hoechst dye and images were acquired, as descried in Materials and Methods. Cultures summarized in this figure are derived from different experimental setups, each including an infected nontreated control culture, which showed viral spread comparable to that in the depicted representative culture. *Culture was terminated, or infection data not recorded, due to virus induced cell death. **Culture was terminated due to drug induced cytotoxicity, possibly enhanced by viral infection.

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Immunostaining, Derivative Assay

Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: VeroE6 cells seeded the previous day in T25 flasks were infected with SARS-CoV-2 followed by treatment with 1-, 1.5-, 2-, 2.5-, 3- and 5-fold EC50 boceprevir, which was administered immediately after infection and subsequently at the indicated timepoints when cells were split, as described in Materials and Methods. Left panel, the % of SARS-CoV-2 infected cells on the specified days post infection was determined by immunostaining. Right panel, replicate cultures were derived following cell splitting and treatment and immunostained for the SARS-CoV-2 Spike protein and counterstained with Hoechst dye and images were acquired, as described in Materials and Methods. Cultures summarized in this figure are derived from different experimental setups, each including an infected nontreated control culture, which showed viral spread comparable to that in the depicted representative culture. *Culture was terminated, or infection data not recorded, due to virus induced cell death. ** Culture was maintained for a total of 17 days without indication of infection (no observation of single SARS-CoV-2 Spike protein positive cells).

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Immunostaining, Derivative Assay

Journal: bioRxiv

Article Title: Hepatitis C Virus Protease Inhibitors Show Differential Efficacy and Interactions with Remdesivir for Treatment of SARS-CoV-2 in Vitro

doi: 10.1101/2020.12.02.408112

Figure Lengend Snippet: VeroE6 cells seeded the previous day in T25 flasks were infected with SARS-CoV-2 followed by treatment with 0.4-fold EC50 of remdesivir (REM) or 1-fold EC50 of PI boceprevir (BOC) or simeprevir (SIM), or a combination of remdesivir with either PI, including an infected, nontreated culture serving as a positive control for viral spread, as described in Materials and Methods. Treatment was administered immediately after infection and subsequently at the indicated timepoints when cells were split, as described in Materials and Methods. Left panel, the % of SARS-CoV-2 infected cells on the specified days post infection was determined by immunostaining. Right panel, replicate cultures were derived following cell splitting and treatment and immunostained for the SARS-CoV-2 Spike protein and counterstained with Hoechst dye and images were acquired, as described in Materials and Methods. *Culture was terminated, or infection data not recorded, due to virus induced cell death. # Culture was maintained for a total of 15 days without indication of infection (no observation of single SARS-CoV-2 Spike protein positive cells).

Article Snippet: In conclusion, following initial partially contradictory reports suggesting efficacy of selected HCV PI against SARS-CoV-2, we here provide a head-to-head comparison of the efficacy of a panel of clinically relevant HCV PI against SARS-CoV-2, including detailed studies of interaction with remdesivir.

Techniques: Infection, Positive Control, Immunostaining, Derivative Assay

Journal: Asaio Journal

Article Title: A Versatile Hybrid Mock Circulation for Hydraulic Investigations of Active and Passive Cardiovascular Implants

doi: 10.1097/MAT.0000000000000851

Figure Lengend Snippet: Electric analog of the numerical models of the cardiovascular system used in this study. Path 1 (P1) corresponds to the BiVAD test case, path 2 (P2) to the TAH test case, and path 3 (P3) to the TCPC test case. Gray lines indicate the interfaces of the respective implant to the numerical model. The additional control mechanisms for heart rate and maximum elastance are only used for the Fontan circulation based on Granegger et al. BiVAD, biventricular assist device; TAH, total artificial heart; TCPC, total cavopulmonary connection.

Article Snippet: The Donovan mock circulation, which is the best-known conventional system, was developed in 1975 and was recently used to evaluate the SynCardia TAH in vitro .

Techniques:

Journal: Asaio Journal

Article Title: A Versatile Hybrid Mock Circulation for Hydraulic Investigations of Active and Passive Cardiovascular Implants

doi: 10.1097/MAT.0000000000000851

Figure Lengend Snippet: Left: Picture of the HMC during the BiVAD and TAH configuration experiments with the two HeartWare HVADs installed. Right: Picture of the HMC during the Fontan experiments with the three-dimensional-printed TCPC. AoP, aortic pressure; BiVAD, biventricular assist device; HMC, hybrid mock circulation; IVCP, inferior vena cava pressure; LAP, left atrial pressure; LPAP, left pulmonary arterial pressure; LVP, left ventricular pressure; PAP, pulmonary arterial pressure; RAP, right atrial pressure; RPAP, right pulmonary arterial pressure; RVP, right ventricular pressure; SVCP, superior vena cava pressure; TAH, total artificial heart; TCPC, total cavopulmonary connection.

Article Snippet: The Donovan mock circulation, which is the best-known conventional system, was developed in 1975 and was recently used to evaluate the SynCardia TAH in vitro .

Techniques:

Journal: Asaio Journal

Article Title: A Versatile Hybrid Mock Circulation for Hydraulic Investigations of Active and Passive Cardiovascular Implants

doi: 10.1097/MAT.0000000000000851

Figure Lengend Snippet: In-vitro performance of a TAH configuration consisting of two HeartWare HVADs and operating either at a constant speed or with physiologic control during an increase of PVR. The signals of the pump speeds and flows as well as of the LAP, the AoP, the RAP, and the PAP are depicted. AoP, aortic pressure; LAP, left atrial pressure; LVAD, left ventricular assist device; PAP, pulmonary arterial pressure; PVR, pulmonary vascular resistance; RAP, right atrial pressure; RVAD, right ventricular assist device; TAH, total artificial heart.

Article Snippet: The Donovan mock circulation, which is the best-known conventional system, was developed in 1975 and was recently used to evaluate the SynCardia TAH in vitro .

Techniques: In Vitro

Journal: Pharmaceutical Research

Article Title: Transferrin Receptor Binding BBB-Shuttle Facilitates Brain Delivery of Anti-Aβ-Affibodies

doi: 10.1007/s11095-022-03282-2

Figure Lengend Snippet: Indirect ELISA of the four affibodies and bispecific Aβ-antibody fusion protein di-scFv3D6-8D3. Plates were coated with ( a ) 250 nM Aβ protofibrils ( b ) 2 μg/ml mTfR1.

Article Snippet: The four affibody proteins were initially assessed for binding to Aβ protofibrils and mTfR1 in vitro .

Techniques: Indirect ELISA

Journal: Pharmaceutical Research

Article Title: Transferrin Receptor Binding BBB-Shuttle Facilitates Brain Delivery of Anti-Aβ-Affibodies

doi: 10.1007/s11095-022-03282-2

Figure Lengend Snippet: Affinities (K D ) in nM for Aβ-Protofibrils and mTfR1 Determined by Indirect ELISA for the Affibodies and di-scFv3D6-8D3. Mean Fold Difference in K D Between Unlabeled and 125 I-Labeled Affibody and di-scFv3D6-8D3, Towards Aβ-protofibrils and mTfR1 Determined by Indirect ELISA (Values Expressed as Mean ± SD)

Article Snippet: The four affibody proteins were initially assessed for binding to Aβ protofibrils and mTfR1 in vitro .

Techniques: Indirect ELISA, Radioactivity