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ace2 inhibition assay kit  (BPS Bioscience)


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    BPS Bioscience ace2 inhibition assay kit
    <t>RBD-ACE2</t> interaction <t>inhibition</t> assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).
    Ace2 Inhibition Assay Kit, supplied by BPS Bioscience, used in various techniques. Bioz Stars score: 94/100, based on 17 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide"

    Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

    Journal: International Journal of Molecular Sciences

    doi: 10.3390/ijms262110607

    RBD-ACE2 interaction inhibition assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).
    Figure Legend Snippet: RBD-ACE2 interaction inhibition assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).

    Techniques Used: Inhibition, Enzyme-linked Immunosorbent Assay, Incubation, Recombinant, Binding Assay, Negative Control, Concentration Assay, Control, Standard Deviation, Two Tailed Test

    NanoLuc bioreporter assays. ( a ) A schematic illustration for the NanoLuc bioreporter assay to determine SARS-CoV-2 S1 RBD—ACE2 inhibition by A13 or Nat1. ( b ) NanoLuc bioreporter assay indicates that both peptides reduce luciferase complementation to the same extent. The signal value for no peptide control was set to 1.00 (6.30 × 10 5 RLU) and all other values were normalized to it. In cell lysates, A13 and Nat1 both reduced luminescence, indicating inhibition of RBD-ACE2 binding. The negative control peptide had no significant effect. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).
    Figure Legend Snippet: NanoLuc bioreporter assays. ( a ) A schematic illustration for the NanoLuc bioreporter assay to determine SARS-CoV-2 S1 RBD—ACE2 inhibition by A13 or Nat1. ( b ) NanoLuc bioreporter assay indicates that both peptides reduce luciferase complementation to the same extent. The signal value for no peptide control was set to 1.00 (6.30 × 10 5 RLU) and all other values were normalized to it. In cell lysates, A13 and Nat1 both reduced luminescence, indicating inhibition of RBD-ACE2 binding. The negative control peptide had no significant effect. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Techniques Used: Inhibition, Luciferase, Control, Binding Assay, Negative Control, Standard Deviation, Two Tailed Test

    Pseudovirus infectivity assays. ( a ) A schematic illustration for pseudovirus infectivity assay to determine the inhibition ability of the peptides to prevent psuedovirus to enter the cell. ( b ) Pseudovirus infectivity assays indicates that both peptides exhibit similar effects on pseudovirus infected cells. The signal value for pseudovirus infection with no peptide was set to 1.00 and corresponding values were normalized to it. Cells were incubated with pseudovirus in the absence/presence of A13 or Nat1, and infection was measured using a luciferase reporter. Both peptides reduce pseudovirus entry into ACE2-expressing cells. Neither peptide shows any visible effects on the non-peptide sample. Data represents the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).
    Figure Legend Snippet: Pseudovirus infectivity assays. ( a ) A schematic illustration for pseudovirus infectivity assay to determine the inhibition ability of the peptides to prevent psuedovirus to enter the cell. ( b ) Pseudovirus infectivity assays indicates that both peptides exhibit similar effects on pseudovirus infected cells. The signal value for pseudovirus infection with no peptide was set to 1.00 and corresponding values were normalized to it. Cells were incubated with pseudovirus in the absence/presence of A13 or Nat1, and infection was measured using a luciferase reporter. Both peptides reduce pseudovirus entry into ACE2-expressing cells. Neither peptide shows any visible effects on the non-peptide sample. Data represents the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Techniques Used: Infection, Inhibition, Incubation, Luciferase, Expressing, Standard Deviation, Two Tailed Test



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    RBD-ACE2 interaction inhibition assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).

    Journal: International Journal of Molecular Sciences

    Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

    doi: 10.3390/ijms262110607

    Figure Lengend Snippet: RBD-ACE2 interaction inhibition assays. ( a ) Schematic illustration of ELISA experiment to evaluate the ability of the A13 or Nat1 to prevent the interaction of SARS-CoV-2 S1 protein RBD to human receptor ACE2. First the plates were incubated with immobilized RBD. They were then inoculated with peptides prior to the addition of recombinant human receptor ACE2. ( b ) Inhibition of RBD-ACE2 binding via A13 or Nat1. The level of RBD-ACE2 binding was measured by chemiluminescence. The negative control with no peptide representing RBD-ACE2 interaction was set to 1.00 (3.2 × 10 4 RLU) and other values were normalized to this number. To the same extent, at the concentration of 0.1 µg/mL, both peptides reduced ACE2 binding compared to the control. At 1.0 µg/mL, inhibition was further enhanced. The negative control peptide did not exhibit a notable inhibition. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001).

    Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

    Techniques: Inhibition, Enzyme-linked Immunosorbent Assay, Incubation, Recombinant, Binding Assay, Negative Control, Concentration Assay, Control, Standard Deviation, Two Tailed Test

    NanoLuc bioreporter assays. ( a ) A schematic illustration for the NanoLuc bioreporter assay to determine SARS-CoV-2 S1 RBD—ACE2 inhibition by A13 or Nat1. ( b ) NanoLuc bioreporter assay indicates that both peptides reduce luciferase complementation to the same extent. The signal value for no peptide control was set to 1.00 (6.30 × 10 5 RLU) and all other values were normalized to it. In cell lysates, A13 and Nat1 both reduced luminescence, indicating inhibition of RBD-ACE2 binding. The negative control peptide had no significant effect. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Journal: International Journal of Molecular Sciences

    Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

    doi: 10.3390/ijms262110607

    Figure Lengend Snippet: NanoLuc bioreporter assays. ( a ) A schematic illustration for the NanoLuc bioreporter assay to determine SARS-CoV-2 S1 RBD—ACE2 inhibition by A13 or Nat1. ( b ) NanoLuc bioreporter assay indicates that both peptides reduce luciferase complementation to the same extent. The signal value for no peptide control was set to 1.00 (6.30 × 10 5 RLU) and all other values were normalized to it. In cell lysates, A13 and Nat1 both reduced luminescence, indicating inhibition of RBD-ACE2 binding. The negative control peptide had no significant effect. Data represent the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

    Techniques: Inhibition, Luciferase, Control, Binding Assay, Negative Control, Standard Deviation, Two Tailed Test

    Pseudovirus infectivity assays. ( a ) A schematic illustration for pseudovirus infectivity assay to determine the inhibition ability of the peptides to prevent psuedovirus to enter the cell. ( b ) Pseudovirus infectivity assays indicates that both peptides exhibit similar effects on pseudovirus infected cells. The signal value for pseudovirus infection with no peptide was set to 1.00 and corresponding values were normalized to it. Cells were incubated with pseudovirus in the absence/presence of A13 or Nat1, and infection was measured using a luciferase reporter. Both peptides reduce pseudovirus entry into ACE2-expressing cells. Neither peptide shows any visible effects on the non-peptide sample. Data represents the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Journal: International Journal of Molecular Sciences

    Article Title: Artificial Intelligence Reveals Nature: Functional Parallels Between a Designed and a Natural Peptide

    doi: 10.3390/ijms262110607

    Figure Lengend Snippet: Pseudovirus infectivity assays. ( a ) A schematic illustration for pseudovirus infectivity assay to determine the inhibition ability of the peptides to prevent psuedovirus to enter the cell. ( b ) Pseudovirus infectivity assays indicates that both peptides exhibit similar effects on pseudovirus infected cells. The signal value for pseudovirus infection with no peptide was set to 1.00 and corresponding values were normalized to it. Cells were incubated with pseudovirus in the absence/presence of A13 or Nat1, and infection was measured using a luciferase reporter. Both peptides reduce pseudovirus entry into ACE2-expressing cells. Neither peptide shows any visible effects on the non-peptide sample. Data represents the mean value from at least three independent experiments. Error bars represent standard deviation. Statistical significance was determined using the nonparametric two-tailed Student’s t -test (**** p ≤ 0.0001, ** p ≤ 0.01).

    Article Snippet: The ability of the peptides to block the interaction between the RBD and ACE2 was assessed using a commercial ACE2 inhibition assay kit (BPS Bioscience, San Diego, CA, United States, CAT# 79931).

    Techniques: Infection, Inhibition, Incubation, Luciferase, Expressing, Standard Deviation, Two Tailed Test

    (A) Attached SARS-CoV-2 virions on the cell surface were detected by RT-qPCR. Calu-3 cells were pretreated with Gal-9 for one hour, then cells were incubated with SARS-CoV-2 (MOI=0.01) in solutions with or without Gal-9 at the indicated concentrations at 4°C for two hours. Cells were washed three times with PBS and harvested for RNA isolation and RT-qPCR measurement of SARS-CoV-2 N gene expression. (B) Relative infectivity of SARS-2-S pseudotyped virus and VSV-G pseudotyped virus in Calu-3 treated with Gal-9 at the indicated concentrations. Calu-3 cells were exposed to Gal-9 for six hours and then infected with SARS-2-S pseudotyped virus or VSV-G pseudotyped virus in solutions containing Gal-9 at the indicated concentrations. Pseudotyped viral entry was analyzed by luciferase activity 24 hpi. Positive serum predetermined to possess anti-SARS-CoV-2 neutralizing activity was used as a negative control. Luciferase signals obtained in the absence of Gal-9 were used for normalization. (C) Relative infectivity of SARS-2-S pseudotyped virus and VSV-G pseudotyped virus in Calu-3 cells treated with anti-ACE2 Ab at the indicated concentrations. Calu-3 cells pre-incubated with anti-ACE2 Ab (R&D Systems, AF933) at the indicated concentrations, or control antibody (anti-goat IgG (R&D Systems, AB-108-C), 50 μg/ml) were co-administered with SARS-2-S pseudotyped and VSV-G pseudotyped virus. At 24 hpi, pseudotyped viral entry was analyzed by luciferase activity. Luciferase signals obtained using the control antibody (anti-goat IgG, 50 μg/ml) were used for normalization. (D) The effect of anti-ACE2 Ab on Gal-9-enhanced cell entry of SARS-2-S was evaluated by measuring luciferase activity. Calu-3 cells were pretreated with Gal-9 for six hours and then pre-incubated with anti-ACE2 Ab (25 μg/ml) or control antibody for one hour, and cells were inoculated with SARS-2-S pseudotyped or VSV-G pseudotyped virus in a solution containing Gal-9 at the indicated concentrations. At 24 hpi, pseudotyped viral entry was analyzed by luciferase activity. Luciferase signals obtained in the absence of both Gal-9 and anti-ACE2 Ab were used for normalization. Data are representative of the results of three independent experiments (mean ± SEM). Statistical significance was analyzed by t test. p>0.05 [ns], p≤0.05 [*], p≤ 0.01 [**], p≤0.001 [***], p≤0.0001 [****].

    Journal: bioRxiv

    Article Title: Human Galectin-9 Potently Enhances SARS-CoV-2 Replication and Inflammation In Airway Epithelial Cells

    doi: 10.1101/2022.03.18.484956

    Figure Lengend Snippet: (A) Attached SARS-CoV-2 virions on the cell surface were detected by RT-qPCR. Calu-3 cells were pretreated with Gal-9 for one hour, then cells were incubated with SARS-CoV-2 (MOI=0.01) in solutions with or without Gal-9 at the indicated concentrations at 4°C for two hours. Cells were washed three times with PBS and harvested for RNA isolation and RT-qPCR measurement of SARS-CoV-2 N gene expression. (B) Relative infectivity of SARS-2-S pseudotyped virus and VSV-G pseudotyped virus in Calu-3 treated with Gal-9 at the indicated concentrations. Calu-3 cells were exposed to Gal-9 for six hours and then infected with SARS-2-S pseudotyped virus or VSV-G pseudotyped virus in solutions containing Gal-9 at the indicated concentrations. Pseudotyped viral entry was analyzed by luciferase activity 24 hpi. Positive serum predetermined to possess anti-SARS-CoV-2 neutralizing activity was used as a negative control. Luciferase signals obtained in the absence of Gal-9 were used for normalization. (C) Relative infectivity of SARS-2-S pseudotyped virus and VSV-G pseudotyped virus in Calu-3 cells treated with anti-ACE2 Ab at the indicated concentrations. Calu-3 cells pre-incubated with anti-ACE2 Ab (R&D Systems, AF933) at the indicated concentrations, or control antibody (anti-goat IgG (R&D Systems, AB-108-C), 50 μg/ml) were co-administered with SARS-2-S pseudotyped and VSV-G pseudotyped virus. At 24 hpi, pseudotyped viral entry was analyzed by luciferase activity. Luciferase signals obtained using the control antibody (anti-goat IgG, 50 μg/ml) were used for normalization. (D) The effect of anti-ACE2 Ab on Gal-9-enhanced cell entry of SARS-2-S was evaluated by measuring luciferase activity. Calu-3 cells were pretreated with Gal-9 for six hours and then pre-incubated with anti-ACE2 Ab (25 μg/ml) or control antibody for one hour, and cells were inoculated with SARS-2-S pseudotyped or VSV-G pseudotyped virus in a solution containing Gal-9 at the indicated concentrations. At 24 hpi, pseudotyped viral entry was analyzed by luciferase activity. Luciferase signals obtained in the absence of both Gal-9 and anti-ACE2 Ab were used for normalization. Data are representative of the results of three independent experiments (mean ± SEM). Statistical significance was analyzed by t test. p>0.05 [ns], p≤0.05 [*], p≤ 0.01 [**], p≤0.001 [***], p≤0.0001 [****].

    Article Snippet: Assessment of Gal-9 effects on the binding of the SARS-CoV-2 spike to human ACE2 was performed using a commercially available spike-ACE2 binding assay kit (CoviDrop SARS-CoV-2 Spike-ACE2 Binding Activity/Inhibition Assay Kit, EPIGENTEK, D-1005-48) following the protocol provided by the manufacturer.

    Techniques: Quantitative RT-PCR, Incubation, Isolation, Expressing, Infection, Luciferase, Activity Assay, Negative Control

    (A) Representative flow cytometry plot describing the protein levels of ACE2 and TMPRSS2 on the surface of Calu-3 cells treated with Gal-9. Calu-3 cells were treated with Gal-9 at the indicated concentrations for 24 h. Cells were then washed and detached before antibody staining for flow cytometry. (B) Percentages of cells expressing ACE2 or TMPRSS2 at the cell surface, measured using flow cytometry. Data represent the results of three independent experiments (mean ± SEM). (C) The dose-response of SARS-CoV-2 spike-ACE2 binding activity measured by reading the absorbance at the wavelength of 450 nm. Data represent the results of three independent experiments (mean ± SEM). Statistical significance was analyzed by t test. p>0.05 [ns], p ≤ 0.05 [*], p ≤ 0.01 [**], p ≤ 0.001 [***], p ≤ 0.0001 [****].

    Journal: bioRxiv

    Article Title: Human Galectin-9 Potently Enhances SARS-CoV-2 Replication and Inflammation In Airway Epithelial Cells

    doi: 10.1101/2022.03.18.484956

    Figure Lengend Snippet: (A) Representative flow cytometry plot describing the protein levels of ACE2 and TMPRSS2 on the surface of Calu-3 cells treated with Gal-9. Calu-3 cells were treated with Gal-9 at the indicated concentrations for 24 h. Cells were then washed and detached before antibody staining for flow cytometry. (B) Percentages of cells expressing ACE2 or TMPRSS2 at the cell surface, measured using flow cytometry. Data represent the results of three independent experiments (mean ± SEM). (C) The dose-response of SARS-CoV-2 spike-ACE2 binding activity measured by reading the absorbance at the wavelength of 450 nm. Data represent the results of three independent experiments (mean ± SEM). Statistical significance was analyzed by t test. p>0.05 [ns], p ≤ 0.05 [*], p ≤ 0.01 [**], p ≤ 0.001 [***], p ≤ 0.0001 [****].

    Article Snippet: Assessment of Gal-9 effects on the binding of the SARS-CoV-2 spike to human ACE2 was performed using a commercially available spike-ACE2 binding assay kit (CoviDrop SARS-CoV-2 Spike-ACE2 Binding Activity/Inhibition Assay Kit, EPIGENTEK, D-1005-48) following the protocol provided by the manufacturer.

    Techniques: Flow Cytometry, Staining, Expressing, Binding Assay, Activity Assay

    Effect of synthetized derivatives on  ACE2:SARS-CoV-2  Spike (RBD) inhibition.

    Journal: Bioorganic Chemistry

    Article Title: Synthesis of novel calcium channel blockers with ACE2 inhibition and dual antihypertensive/anti-inflammatory effects: A possible therapeutic tool for COVID-19

    doi: 10.1016/j.bioorg.2021.105272

    Figure Lengend Snippet: Effect of synthetized derivatives on ACE2:SARS-CoV-2 Spike (RBD) inhibition.

    Article Snippet: In order to measure Angiotensin-Converting Enzyme 2 (ACE2) receptor inhibition activity, the ACE2:SARS-CoV-2 Spike (RBD) Inhibitor Screening Colorimetric Assay Kit # 78031 (Bps Bioscience, Cornerstone Court W, Ste B San Diego, CA 92121) was used.

    Techniques: Inhibition

    SAR for the most potent compounds. DHPMs: Dihydropyrimidines, CCBs: calcium channel blockers, ACE2: Angiotensin-converting enzyme 2, THP-1: Human monocytic cell line, IL-6: Interleukine-6, LPS: Lipopolysaccharide, CRP: C-Reactive Protein.

    Journal: Bioorganic Chemistry

    Article Title: Synthesis of novel calcium channel blockers with ACE2 inhibition and dual antihypertensive/anti-inflammatory effects: A possible therapeutic tool for COVID-19

    doi: 10.1016/j.bioorg.2021.105272

    Figure Lengend Snippet: SAR for the most potent compounds. DHPMs: Dihydropyrimidines, CCBs: calcium channel blockers, ACE2: Angiotensin-converting enzyme 2, THP-1: Human monocytic cell line, IL-6: Interleukine-6, LPS: Lipopolysaccharide, CRP: C-Reactive Protein.

    Article Snippet: In order to measure Angiotensin-Converting Enzyme 2 (ACE2) receptor inhibition activity, the ACE2:SARS-CoV-2 Spike (RBD) Inhibitor Screening Colorimetric Assay Kit # 78031 (Bps Bioscience, Cornerstone Court W, Ste B San Diego, CA 92121) was used.

    Techniques: