Bacteria:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities.
Article Snippet: Antimicrobial Susceptibility Assay against ESKAPEE Pathogens. (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2−8 × 105 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii; and MIC was additionally confirmed by OD600. (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar https://doi.org/10.1021/acs.jmedchem.4c00912 J. Med. ..
Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Concentration Assay:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities.
Article Snippet: Antimicrobial Susceptibility Assay against ESKAPEE Pathogens. (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2−8 × 105 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii; and MIC was additionally confirmed by OD600. (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar https://doi.org/10.1021/acs.jmedchem.4c00912 J. Med. ..
Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Incubation:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities.
Article Snippet: Antimicrobial Susceptibility Assay against ESKAPEE Pathogens. (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2−8 × 105 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii; and MIC was additionally confirmed by OD600. (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar https://doi.org/10.1021/acs.jmedchem.4c00912 J. Med. ..
Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Generated:Article Title: Detection of Human Intestinal Catalase-Negative, Gram-Positive Cocci by rRNA-Targeted Reverse Transcription-PCR
Article Snippet: .. Each analytical curve ranging from 103 or 104 to 109 cells per 10 ml was generated with RT-qPCR data, threshold cycle (CT) value, and the corresponding cell count, which was determined microscopically with the DAPI staining method, of the dilution series of the following representative strains: E. faecalis ATCC 19433T (for g-Encoc-F/R and s-Efs-F/R), E. caccae DSM 19114T (for s-EcaccF/R), E. cecorum JCM 8724T (for s-Ececo-F/R), E. sulfureus DSM 6905T (for sg-Esulf-F/R), E. casseliflavus JCM 8723T (for sg-Ecass-F/R), E. avium JCM 8722T (for sg-Eavi-F/R), E. dispar DSM 6630T (for s-Edis-F/R), E. faecium ATCC 19434T (for sg-Efm-F/R and s-Efm-F/R), S. salivarius JCM 5707T (for g-Str-F/R and s-Ssal-F/R), S. mitis GIFU 12458T (for s-Spn-F/R), S. agalactiae JCM 5671T (for s-Sag-F/R), S. pyogenes ATCC 12344T (for s-Spy-F/R), L. lactis subsp. lactis ATCC 19435T (for sg-Lclac-F/R) and L. piscium DSM 6634T (for sg-Lcpis-F/R). ..
Quantitative RT-PCR:Article Title: Detection of Human Intestinal Catalase-Negative, Gram-Positive Cocci by rRNA-Targeted Reverse Transcription-PCR
Article Snippet: .. Each analytical curve ranging from 103 or 104 to 109 cells per 10 ml was generated with RT-qPCR data, threshold cycle (CT) value, and the corresponding cell count, which was determined microscopically with the DAPI staining method, of the dilution series of the following representative strains: E. faecalis ATCC 19433T (for g-Encoc-F/R and s-Efs-F/R), E. caccae DSM 19114T (for s-EcaccF/R), E. cecorum JCM 8724T (for s-Ececo-F/R), E. sulfureus DSM 6905T (for sg-Esulf-F/R), E. casseliflavus JCM 8723T (for sg-Ecass-F/R), E. avium JCM 8722T (for sg-Eavi-F/R), E. dispar DSM 6630T (for s-Edis-F/R), E. faecium ATCC 19434T (for sg-Efm-F/R and s-Efm-F/R), S. salivarius JCM 5707T (for g-Str-F/R and s-Ssal-F/R), S. mitis GIFU 12458T (for s-Spn-F/R), S. agalactiae JCM 5671T (for s-Sag-F/R), S. pyogenes ATCC 12344T (for s-Spy-F/R), L. lactis subsp. lactis ATCC 19435T (for sg-Lclac-F/R) and L. piscium DSM 6634T (for sg-Lcpis-F/R). ..
Cell Characterization:Article Title: Detection of Human Intestinal Catalase-Negative, Gram-Positive Cocci by rRNA-Targeted Reverse Transcription-PCR
Article Snippet: .. Each analytical curve ranging from 103 or 104 to 109 cells per 10 ml was generated with RT-qPCR data, threshold cycle (CT) value, and the corresponding cell count, which was determined microscopically with the DAPI staining method, of the dilution series of the following representative strains: E. faecalis ATCC 19433T (for g-Encoc-F/R and s-Efs-F/R), E. caccae DSM 19114T (for s-EcaccF/R), E. cecorum JCM 8724T (for s-Ececo-F/R), E. sulfureus DSM 6905T (for sg-Esulf-F/R), E. casseliflavus JCM 8723T (for sg-Ecass-F/R), E. avium JCM 8722T (for sg-Eavi-F/R), E. dispar DSM 6630T (for s-Edis-F/R), E. faecium ATCC 19434T (for sg-Efm-F/R and s-Efm-F/R), S. salivarius JCM 5707T (for g-Str-F/R and s-Ssal-F/R), S. mitis GIFU 12458T (for s-Spn-F/R), S. agalactiae JCM 5671T (for s-Sag-F/R), S. pyogenes ATCC 12344T (for s-Spy-F/R), L. lactis subsp. lactis ATCC 19435T (for sg-Lclac-F/R) and L. piscium DSM 6634T (for sg-Lcpis-F/R). ..
Staining:Article Title: Detection of Human Intestinal Catalase-Negative, Gram-Positive Cocci by rRNA-Targeted Reverse Transcription-PCR
Article Snippet: .. Each analytical curve ranging from 103 or 104 to 109 cells per 10 ml was generated with RT-qPCR data, threshold cycle (CT) value, and the corresponding cell count, which was determined microscopically with the DAPI staining method, of the dilution series of the following representative strains: E. faecalis ATCC 19433T (for g-Encoc-F/R and s-Efs-F/R), E. caccae DSM 19114T (for s-EcaccF/R), E. cecorum JCM 8724T (for s-Ececo-F/R), E. sulfureus DSM 6905T (for sg-Esulf-F/R), E. casseliflavus JCM 8723T (for sg-Ecass-F/R), E. avium JCM 8722T (for sg-Eavi-F/R), E. dispar DSM 6630T (for s-Edis-F/R), E. faecium ATCC 19434T (for sg-Efm-F/R and s-Efm-F/R), S. salivarius JCM 5707T (for g-Str-F/R and s-Ssal-F/R), S. mitis GIFU 12458T (for s-Spn-F/R), S. agalactiae JCM 5671T (for s-Sag-F/R), S. pyogenes ATCC 12344T (for s-Spy-F/R), L. lactis subsp. lactis ATCC 19435T (for sg-Lclac-F/R) and L. piscium DSM 6634T (for sg-Lcpis-F/R). ..
Suspension:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Control:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Mutagenesis:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
Cell Culture:Article Title: Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
Article Snippet: (Protocol a) Antimicrobial activity test against ESKAPEE strains S. aureus NRS 1, K. pneumoniae ATCC 700603, A. baumannii GSAB 164, P. aeruginosa NCTC 13437, and E. coli ATCC 25922 was purchased from Evotec (UK). .. The protocol was similar to only a few exceptions: E. faecium was recovered on Columbia blood agar; buffer mixture of 1% (v/v) DMSO and 0.1 M PBS; bacteria ∼2–8 × 10 5 CFU/mL; round-bottom well plate; the highest tested concentration of peptides ∼15 μM; incubation for 18 and 20 h for A. baumannii ; and MIC was additionally confirmed by OD 600 . (Protocol b) Test against ESKAPEE strains E. faecalis JH2-2 and UCN41, E. faecium ATCC 19434T and BM4147, S. aureus ATCC 25923, and A. baumannii CIP7010, was performed using a similar protocol with only a few exceptions: cation-adjusted MHB was used; bacterial inoculum was prepared directly from the 0.5 McFarland (McF)-adjusted colony suspension, diluted 1:20 to a final concentration of 5 × 10 5 CFU/mL; the highest tested concentration of peptides ∼30 μM; incubation for 16 to 20 h at 35 ± 2 °C; and the quality control strain of P. aeruginosa (ATCC 27853) was used as the control using ceftazidime as the reference antibiotic. (Protocol c) Test against ESKAPEE strains E. faecium Z906, S. aureus ATCC 29213, S. epidermidis 30 WT, K. pneumoniae E1120, E1267, and 4371, A. baumannii Z13, P. aeruginosa K11 and K12 and E. coli E1098, S. aureus protease mutants: NE1506, NE934, NE163, and NE1740, protease null mutant ANG 2038 (AH1919) and control strains ANG 1575 (AH1263), and S. aureus USA 300 derivative JE2 was performed using a similar protocol with only a few exceptions: bacteria were cultured for 18 h; several colonies were scraped and resuspended in 0.9% NaCl to 10 8 CFU/mL (0.5 McF); the highest tested concentration of peptides ∼30 μM (128 mg/L); the MIC was considered as the concentration that reduced ≥80% bacterial growth (OD 600 ) compared to control. (Protocol d) Test against ESKAPEE strains E. faecium ATCC 700221 (vancomycin-resistant), E. faecalis ATCC 700802 (vancomycin-resistant), S. aureus ATCC 12600 and ATCC BAA-1556 (methicillin-resistant), K. pneumoniae ATCC 13883, A. baumannii ATCC 19606, P. aeruginosa PAO1 and PA14, E. coli ATCC 11775, and AIC221 and AIC222 (colistin-resistant) was performed using a similar protocol with only a few exceptions: bacteria ∼2 × 10 6 cells mL –1 , Luria–Bertani (LB) medium, and 20 h incubation before the visual assessment of MIC. ..
other:
Sequencing:Article Title: Enterococcus Xinjiangensis sp. nov., Isolated from Yogurt of Xinjiang, China.
Article Snippet: A Gram-strain-positive bacterial strain 48 was isolated from traditional yogurt in Xinjiang Province, China.. The bacterium was characterized by a polyphasic approach, including 16S rRNA gene sequence analysis, polymerase a subunit (rpoA) gene sequence analysis, determination of DNA G?C content, DNA–DNA hybridization with the type strain of Enterococcus ratti and analysis of phenotypic features.. Strain 48 accounted for 96.1, 95.8, 95.8, and 95.7 % with Enterococcus faecium CGMCC 1.2136, Enterococcus hirae ATCC 9790, Enterococcus durans CECT 411, and E. ratti ATCC 700914 in the 16S rRNA gene sequence similarities, respectively.
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