Journal: mBio

Article Title: Peptidyl tRNA Hydrolase Is Required for Robust Prolyl-tRNA Turnover in Mycobacterium tuberculosis

doi: 10.1128/mbio.03469-22

Figure Lengend Snippet: Pth depletion sensitizes Mtb to macrolides. (A) Mtb growth in the presence of two macrolides, erythromycin and clarithromycin, is plotted across drug concentrations as determined by fluorescence in an alamarBlue assay. Briefly, antibiotic-containing plates were incubated with Mtb cells for 6 days, at which point resazurin was added, followed by 48 h of additional agitation at 37°C. Fluorescence was normalized to the OD 600 and to the positive control for each strain (no antibiotic). The fractions of bacteria surviving relative to the no-drug control (“Normalized fraction surviving”) are plotted against drug concentrations, along with a least-squares fit of the dose response . The Pth CRISPRi strain was grown to mid-log phase, diluted to an OD 600 of 0.001, and plated with serial dilutions of antibiotics in 96-well plates. The fraction of Mtb cells surviving is plotted by normalizing the fluorescence values to the values for the control wells with no drug. Means with standard errors from three biological replicates are shown. (B) Cu-tRNAseq was performed on Pth CRISPRi strains in the presence and absence of erythromycin. Strains were grown in the presence or absence of the inducer as described in the legend of Fig. 3 . Uninduced strains were then treated with 25 μg/mL of erythromycin overnight, and Pth knockdown-induced strains were treated with 2 μg/mL for the same duration. Means with standard errors from three biological replicates are shown.

Article Snippet: Briefly, strains were diluted to an OD 600 of 0.001 and tested in technical duplicate using serial dilutions of the following antibiotics: erythromycin (GoldBio), clarithromycin (GoldBio), puromycin (GoldBio), clindamycin (GoldBio), chloramphenicol (Sigma-Aldrich), isoniazid (Sigma-Aldrich), ethambutol (Sigma-Aldrich), kanamycin (IBI Scientific), fusidic acid (Sigma-Aldrich), linezolid (Sigma-Aldrich), and capreomycin (Sigma-Aldrich).

Techniques: Fluorescence, Alamar Blue Assay, Incubation, Positive Control, Bacteria, Control, Knockdown

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein-Based Strategy to Reduce Antibiotic Dosing for the Treatment of Pneumococcal Infection

doi: 10.3389/fimmu.2020.620784

Figure Lengend Snippet: Survival of mice infected with pneumococci and treated with different doses of clarithromycin. Clarithromycin was injected four times, at 12, 36, 60 and 84 h, after the administration of pneumococci. Six mice were used for each dose of clarithromycin.

Article Snippet: The antibiotic clarithromycin (Santa Cruz Biotechnology, sc-205634) was reconstituted in acetone at a concentration of 50 mg/ml and stored at 4°C for a maximum of 5 days.

Techniques: Infection, Injection

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein-Based Strategy to Reduce Antibiotic Dosing for the Treatment of Pneumococcal Infection

doi: 10.3389/fimmu.2020.620784

Figure Lengend Snippet: Survival of mice infected with pneumococci and treated with E-CRP-1 and clarithromycin. E-CRP-1 or WT CRP was injected 12 h after administering pneumococci and is indicated by an arrow on the x-axis. Clarithromycin (0.02 mg) was injected four times, at 13, 36, 60 and 84 h, after the administration of pneumococci. The data are combined from two separate experiments with six to eight mice in each group in each experiment. The p -values for the differences in the survival curves between groups A B and A C were 0.004 and 0.006, respectively. The p -value for the difference in the survival curves between groups B and C was 0.94. The p -values for the differences in the survival curves between groups C D and C E were 0.23 and <0.001, respectively. The p -values for the differences in the survival curves between groups B E and D E were <0.001.

Article Snippet: The antibiotic clarithromycin (Santa Cruz Biotechnology, sc-205634) was reconstituted in acetone at a concentration of 50 mg/ml and stored at 4°C for a maximum of 5 days.

Techniques: Infection, Injection

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein-Based Strategy to Reduce Antibiotic Dosing for the Treatment of Pneumococcal Infection

doi: 10.3389/fimmu.2020.620784

Figure Lengend Snippet: Bacteremia in mice infected with pneumococci and treated with E-CRP-1 and clarithromycin. Blood was collected from each surviving mouse shown in Figure 2 . (A) Scatter plots of the bacteremia data. The horizontal red line in each group of mice represents median bacteremia. (B) The median bacteremia values, derived from (A) For 36–92 h, the p -value for the difference between groups A and B was <0.05. For 20–68 h, the p -value for the difference between groups A and C was <0.05. For all time points, the p -values for the differences between groups B C and C D were >0.05. For 36–116 h, the p -values for the differences between groups C E, B E, and D E were <0.05, with most significant difference ( p < 0.005) between 44–92 h.

Article Snippet: The antibiotic clarithromycin (Santa Cruz Biotechnology, sc-205634) was reconstituted in acetone at a concentration of 50 mg/ml and stored at 4°C for a maximum of 5 days.

Techniques: Infection, Derivative Assay

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein-Based Strategy to Reduce Antibiotic Dosing for the Treatment of Pneumococcal Infection

doi: 10.3389/fimmu.2020.620784

Figure Lengend Snippet: Survival of mice infected with pneumococci and treated with E-CRP-2 and clarithromycin. E-CRP-2 was injected 12 h after administering pneumococci and is indicated by an arrow on the x-axis. Clarithromycin (0.02 mg) was injected four times, at 13, 36, 60 and 84 h, after the administration of pneumococci. The data are combined from two separate experiments with six to eight mice in each group in each experiment. The p -values for the differences in the survival curves between groups A B and A C were <0.001 and 0.002, respectively. The p -value for the difference in the survival curves between groups B and C was 0.25. The p -values for the differences in the survival curves between groups B D and C D were 0.01 and 0.002, respectively.

Article Snippet: The antibiotic clarithromycin (Santa Cruz Biotechnology, sc-205634) was reconstituted in acetone at a concentration of 50 mg/ml and stored at 4°C for a maximum of 5 days.

Techniques: Infection, Injection

Journal: Frontiers in Immunology

Article Title: C-Reactive Protein-Based Strategy to Reduce Antibiotic Dosing for the Treatment of Pneumococcal Infection

doi: 10.3389/fimmu.2020.620784

Figure Lengend Snippet: Bacteremia in mice infected with pneumococci and treated with E-CRP-2 and clarithromycin. Blood was collected from each surviving mouse shown in Figure 4 . (A) Scatter plots of the bacteremia data. The horizontal red line in each group of mice represents median bacteremia. (B) The median bacteremia values, derived from (A) For 36–116 h, the p -value for the difference between groups A and B was <0.005. The p -values for the difference between groups A and C was <0.005 and <0.05 for 44–68 h and 92–116 h, respectively. For all time points, the p -values for the differences between groups B C and B D were >0.05. For 60 to 116 h, the p -value for the difference between groups C and D was <0.05.

Article Snippet: The antibiotic clarithromycin (Santa Cruz Biotechnology, sc-205634) was reconstituted in acetone at a concentration of 50 mg/ml and stored at 4°C for a maximum of 5 days.

Techniques: Infection, Derivative Assay

Journal: Circulation

Article Title: CAVIN1-Mediated hERG Dynamics: A Novel Mechanism Underlying the Interindividual Variability in Drug-Induced Long QT

doi: 10.1161/circulationaha.123.063917

Figure Lengend Snippet: Figure 6. Universal mechanism implicating CAVIN1 in response to other hERG blockers. A, Top, Representative superimposed noncorrected field potential (FP) recorded from low-sensitivity (LS) and high-sensitivity (HS) induced pluripotent stem cell–derived cardiomyocytes (iPS-CMs) after treatment with different concentrations of E4031. Bottom: averaged percentage of change in corrected field potential duration (FPDc; corrected to beating frequency) compared with that at baseline after application of 0.1 and 1 µM E4031. *P<0.05 and ***P<0.001 vs LS iPS-CMs (mixed effect models). B, Top, Aligned noncorrected FP recorded from LS and HS iPS-CMs treated with different concentrations of vandetanib. Bottom: averaged percentage of change in FPDc (corrected to beating frequency) compared with that at baseline after application of 0.1, 1, and 10 µM of vandetanib (mixed effect models). C, Top, Aligned noncorrected FP recorded from LS and HS iPS-CMs treated with different concentrations of the antibiotic clarithromycin. Bottom, Averaged percentage of change in FPDc (corrected to beating frequency) compared with that at baseline after application of 10, 30, and 100 µM of clarithromycin. D through F, Averaged percentage of change in FPDc compared with that at baseline measured in LS iPS-CMs infected with either GFP or CAVIN1-T2A GFP adenoviruses in response to increasing concentrations of E4031, vandetanib, and clarithromycin. *P<0.05 vs GFP-infected LS iPS-CMs, respectively (mixed effect models). G through I, Percentage change in FPDc compared with that at baseline after application of different concentrations of (from left to right) E4031, vandetanib, and clarithromycin at day 5 of transfection of HS iPS-CMs with either siNeg or CAVIN1 small interfering RNA. *P<0.05 vs GFP-infected LS iPS-CMs, respectively (mixed effect models).

Article Snippet: Each drug (sotalol [Selleckchem], E4031 [StressMarq], clarithromycin [Selleckchem], and vandetanib [Selleckchem]) was prepared at different concentrations in RPMI medium supplemented with B27 and insulin and directly added to wells during recording at 37 °C.

Techniques: Derivative Assay, Infection, Transfection, Small Interfering RNA

Journal: Polymers

Article Title: Preparation and Application of Molecularly Imprinted Monolithic Extraction Column for the Selective Microextraction of Multiple Macrolide Antibiotics from Animal Muscles

doi: 10.3390/polym11071109

Figure Lengend Snippet: Specific adsorption capacities of MIMC and NIMC for single and multiple macrolides a .

Article Snippet: ROX, SPM, clarithromycin (CLA), and tulathromycin (TUL) were purchased from European Pharmacopoeia (EDQM, Strasbourg, France).

Techniques: Adsorption, Polymer

Journal: Polymers

Article Title: Preparation and Application of Molecularly Imprinted Monolithic Extraction Column for the Selective Microextraction of Multiple Macrolide Antibiotics from Animal Muscles

doi: 10.3390/polym11071109

Figure Lengend Snippet: Effects of ( A ) methanol (MeOH), acetonitrile (ACN) and ethyl acetate (EA) as loading solvents and ( B ) MeOH, ACN, acetone and water as washing solvents on the recoveries of six macrolide drugs: erythromycin (ERY); clarithromycin (CLA); azithromycin (AZI); tulathromycin (TUL); tilmicosin (TIM); and, spiramycin (SPM).

Article Snippet: ROX, SPM, clarithromycin (CLA), and tulathromycin (TUL) were purchased from European Pharmacopoeia (EDQM, Strasbourg, France).

Techniques:

Journal: Polymers

Article Title: Preparation and Application of Molecularly Imprinted Monolithic Extraction Column for the Selective Microextraction of Multiple Macrolide Antibiotics from Animal Muscles

doi: 10.3390/polym11071109

Figure Lengend Snippet: Validation data for six macrolide antibiotics after molecularly imprinted polymer monolith microextraction (MIPMME) procedure in spiked animal muscle samples a .

Article Snippet: ROX, SPM, clarithromycin (CLA), and tulathromycin (TUL) were purchased from European Pharmacopoeia (EDQM, Strasbourg, France).

Techniques: Biomarker Discovery, Polymer