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(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . <t>trachomatis</t> growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .
C Trachomatis E Bour, supplied by DSMZ, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . <t>trachomatis</t> growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .
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(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . <t>trachomatis</t> growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .
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(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . <t>trachomatis</t> growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .
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(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . <t>trachomatis</t> growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .
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Image Search Results


(A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .

Journal: PLOS Biology

Article Title: A multi-strategy antimicrobial discovery approach reveals new ways to treat Chlamydia

doi: 10.1371/journal.pbio.3003123

Figure Lengend Snippet: (A) Chemical diversity of the screening library of 36,785 compounds, illustrated by principal component analysis based on molecular descriptors calculated with PaDEL-Descriptor . “Drugs” denotes 6,798 compounds from the Drug Repurposing Hub , included to represent the chemical space of pharmaceutical drugs. (B) Screening protocol, highlighting major experimental steps, time points, and measurements. Compounds were tested at 10 µM. (C) Bulk fluorescence measurements of GFP and resorufin, representing C . trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (D) Image-based measurements of inclusion area and count and host cell nuclei count, representing C. trachomatis growth and host cell viability, respectively. The dashed lines indicate the cut-offs used for hit selection. (E) Representative image examples of a non-hit and a hit, as well as positive (ciprofloxacin, CIP) and negative (DMSO vehicle) controls for bacterial growth inhibition. Scale bar is 10 µm. (F) The 271 hits from the screen retested in duplicate with the screening assay protocol at 1, 5, and 10 µM, highlighting the overall consistency between the two replicates. (G) Bacterial growth inhibition achieved by the 271 hit compounds, shown at the single-compound level. The compounds are sorted by mean bulk fluorescence across the three concentrations. (H) Estimated IC 50 of the 271 hits. (I) Maximal structural similarity, represented by Tanimoto coefficients, between the hits and 506 known antibiotics in the Drug Repurposing Hub . The dashed line indicates the maximum value among the compounds prioritized. (J) Potency of the 52 priority compounds, determined with the screening assay protocol. (K) Dose–response curves of the four most potent compounds identified by experimental screening (c1 e –c4 e ) (mean ± SD, n = 3). Lines (green, GFP fluorescence; brown, inclusion area; gray, bacterial genome copies) indicate curve fits used for IC 50 calculation. (L) Chemical structures of the four most potent compounds, drawn based on their SMILES strings using OpenBabel (version 3.0.0) . The data underlying this figure can be found in .

Article Snippet: Infection experiments were carried out with the following Chlamydia strains: C. trachomatis L2/434/Bu (CTL2, ATCC VR-902B), an rsGFP-expressing derivative of CTL2 (CTL2-GFP; i.e., CTL2 transformed with plasmid p2TK2-SW2-IncDProm-RSGFP-IncDTerm [ ]), C. trachomatis A/HAR-13 (CTA; DSMZ, 19440), C. trachomatis D/UW-3/Cx (CTD; DSMZ, 19411), C. trachomatis E/Bour (CTE; DSMZ, 19131), C. caviae GPIC (CC; [ ]), and C. muridarum MoPn (CM; DSMZ, 28544).

Techniques: Fluorescence, Selection, Inhibition, Screening Assay