carbenicillin Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 93
    Millipore carbenicillin
    Genetic dominance limits the phenotypic contribution of horizontally transferred recessive alleles. (A) Pictures of a representative replicate of the conjugation assays. Overnight cultures of spots inoculated from 10-fold dilutions of conjugation mixes (10 0 to 10 −4 , from left to right) on plates selecting for transconjugants. Selection on <t>carbenicillin</t> reveals the actual number of transconjugants; selection on carbenicillin plus nalidixic acid or trimethoprim reveals the number of transconjugants carrying gyrA and folA alleles and expressing the resistant phenotype. (B) Antibiotic resistance level conferred by a plasmid-encoded resistant allele in the recipient bacterium (when a wild-type copy of the gene is present in the chromosome), assuming phenotypic resistance as the product of plasmid copy number and the coefficient of dominance of the allele. Experimental data are presented for gyrA D87N and folA L28R in plasmid pSEVA121.
    Carbenicillin, supplied by Millipore, used in various techniques. Bioz Stars score: 93/100, based on 2075 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/Millipore
    Average 93 stars, based on 2075 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    93/100 stars
      Buy from Supplier

    90
    Millipore carbenicillin disodium salt
    SLC25A1 genetic defects confer susceptibility to mitochondrial proteostasis inhibitors. Oxygen consumption rate (OCR) was analyzed with Seahorse technology in wild type and two clones of SLC25A1 null cells. Cells were incubated in the absence or presence of drugs that target mitochondrial protein synthesis (A-I) or mitochondrial protein quality control (J). <t>Carbenicillin</t> was used as a control drug (O, n=3). All cells were treated for 48h with drugs except for panel A. A) Depicts a stress test after increasing incubation times with 2.25 µM doxycycline. Average ± SD n=8. Arrows A to C mark times of stress test drug injections: oligomycin, FCCP, and antimycin plus rotenone; respectively. B-D) and E-G) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline or minocycline treated SLC25A1 null and wild type cells. Average ± SE, n=4-6. H-J) Basal OCR of cells treated in the absence or presence of chloramphenicol, linezolid, or actinonin. Average ± SE, n=4-7. K-O) Basal OCR ratios for drug treated and untreated wild type and two SLC25A1 null cell clones (Δ and Δ1). P) Same as A, but wild type and SLC25A4 null cells were treated for 48h in the absence or presence of 2.25 µM doxycycline. Q) Basal OCR ratios for drug treated and untreated wild type and SLC25A4 null cells. R) Crystal violet cell survival assay for DU145 and ρ0 cells in the absence or presence of drug. Average ± SE, n=3. S) Same as A but with DU145 and ρ0 cells. DU145 cells treated in the presence of 2.25 µM doxycycline are shown by gray square symbols. Average ± SE, n=3. T-V) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline treated DU145 and ρ0 cells. Average ± SE, n=3. For A-J p values were calculated with Two-Factor ANOVA with Repeated Measures. For K-O and Q p values were calculated by One Way ANOVA followed by multiple comparison correction by the Bonferroni method. For R, p values were calculated One Way ANOVA followed by Fisher’s Least Significant Difference Comparison Test. In panels A-V, wild type and mutants are marked by gray and blue symbols, respectively.
    Carbenicillin Disodium Salt, supplied by Millipore, used in various techniques. Bioz Stars score: 90/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin disodium salt/product/Millipore
    Average 90 stars, based on 35 article reviews
    Price from $9.99 to $1999.99
    carbenicillin disodium salt - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    90
    Thermo Fisher carbenicillin
    SLC25A1 genetic defects confer susceptibility to mitochondrial proteostasis inhibitors. Oxygen consumption rate (OCR) was analyzed with Seahorse technology in wild type and two clones of SLC25A1 null cells. Cells were incubated in the absence or presence of drugs that target mitochondrial protein synthesis (A-I) or mitochondrial protein quality control (J). <t>Carbenicillin</t> was used as a control drug (O, n=3). All cells were treated for 48h with drugs except for panel A. A) Depicts a stress test after increasing incubation times with 2.25 µM doxycycline. Average ± SD n=8. Arrows A to C mark times of stress test drug injections: oligomycin, FCCP, and antimycin plus rotenone; respectively. B-D) and E-G) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline or minocycline treated SLC25A1 null and wild type cells. Average ± SE, n=4-6. H-J) Basal OCR of cells treated in the absence or presence of chloramphenicol, linezolid, or actinonin. Average ± SE, n=4-7. K-O) Basal OCR ratios for drug treated and untreated wild type and two SLC25A1 null cell clones (Δ and Δ1). P) Same as A, but wild type and SLC25A4 null cells were treated for 48h in the absence or presence of 2.25 µM doxycycline. Q) Basal OCR ratios for drug treated and untreated wild type and SLC25A4 null cells. R) Crystal violet cell survival assay for DU145 and ρ0 cells in the absence or presence of drug. Average ± SE, n=3. S) Same as A but with DU145 and ρ0 cells. DU145 cells treated in the presence of 2.25 µM doxycycline are shown by gray square symbols. Average ± SE, n=3. T-V) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline treated DU145 and ρ0 cells. Average ± SE, n=3. For A-J p values were calculated with Two-Factor ANOVA with Repeated Measures. For K-O and Q p values were calculated by One Way ANOVA followed by multiple comparison correction by the Bonferroni method. For R, p values were calculated One Way ANOVA followed by Fisher’s Least Significant Difference Comparison Test. In panels A-V, wild type and mutants are marked by gray and blue symbols, respectively.
    Carbenicillin, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 444 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/Thermo Fisher
    Average 90 stars, based on 444 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    92
    BioShop carbenicillin
    SLC25A1 genetic defects confer susceptibility to mitochondrial proteostasis inhibitors. Oxygen consumption rate (OCR) was analyzed with Seahorse technology in wild type and two clones of SLC25A1 null cells. Cells were incubated in the absence or presence of drugs that target mitochondrial protein synthesis (A-I) or mitochondrial protein quality control (J). <t>Carbenicillin</t> was used as a control drug (O, n=3). All cells were treated for 48h with drugs except for panel A. A) Depicts a stress test after increasing incubation times with 2.25 µM doxycycline. Average ± SD n=8. Arrows A to C mark times of stress test drug injections: oligomycin, FCCP, and antimycin plus rotenone; respectively. B-D) and E-G) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline or minocycline treated SLC25A1 null and wild type cells. Average ± SE, n=4-6. H-J) Basal OCR of cells treated in the absence or presence of chloramphenicol, linezolid, or actinonin. Average ± SE, n=4-7. K-O) Basal OCR ratios for drug treated and untreated wild type and two SLC25A1 null cell clones (Δ and Δ1). P) Same as A, but wild type and SLC25A4 null cells were treated for 48h in the absence or presence of 2.25 µM doxycycline. Q) Basal OCR ratios for drug treated and untreated wild type and SLC25A4 null cells. R) Crystal violet cell survival assay for DU145 and ρ0 cells in the absence or presence of drug. Average ± SE, n=3. S) Same as A but with DU145 and ρ0 cells. DU145 cells treated in the presence of 2.25 µM doxycycline are shown by gray square symbols. Average ± SE, n=3. T-V) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline treated DU145 and ρ0 cells. Average ± SE, n=3. For A-J p values were calculated with Two-Factor ANOVA with Repeated Measures. For K-O and Q p values were calculated by One Way ANOVA followed by multiple comparison correction by the Bonferroni method. For R, p values were calculated One Way ANOVA followed by Fisher’s Least Significant Difference Comparison Test. In panels A-V, wild type and mutants are marked by gray and blue symbols, respectively.
    Carbenicillin, supplied by BioShop, used in various techniques. Bioz Stars score: 92/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/BioShop
    Average 92 stars, based on 4 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    92
    Carl Roth GmbH carbenicillin
    Collateral sensitivity. Validated collateral sensitivity for 38 distinct populations obtained from Barbosa et al. (2017) . Six to seven populations resistant to CIP, ( a ); STR, ( b ); GEN, ( c ); PIT, ( d ); CAR, ( e ); and CEF, ( f ), were tested for collaterals sensitivity against 2 or four drugs, indicated above each of the panels. The ancestor PA14 is shown in black and the various populations in distinct colors. Points indicate averages of 3 technical replicates per population and drug concentration. CAR, <t>carbenicillin;</t> GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; CIP, ciprofloxacin; and CEF, cefsulodin; superscript R denotes resistance against the particular drug. The data for this figure is provided in Figure 5—figure supplement 1—source data 1 . Dose-response curves data of surviving populations in the generalized experiment. The data is shown in Figure 5—figure supplement 1 and summarized in Figure 5 . Optical density values were recorded after 12 hr of incubation at 37 °C.
    Carbenicillin, supplied by Carl Roth GmbH, used in various techniques. Bioz Stars score: 92/100, based on 108 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/Carl Roth GmbH
    Average 92 stars, based on 108 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    92/100 stars
      Buy from Supplier

    90
    Cellgro carbenicillin
    Collateral sensitivity. Validated collateral sensitivity for 38 distinct populations obtained from Barbosa et al. (2017) . Six to seven populations resistant to CIP, ( a ); STR, ( b ); GEN, ( c ); PIT, ( d ); CAR, ( e ); and CEF, ( f ), were tested for collaterals sensitivity against 2 or four drugs, indicated above each of the panels. The ancestor PA14 is shown in black and the various populations in distinct colors. Points indicate averages of 3 technical replicates per population and drug concentration. CAR, <t>carbenicillin;</t> GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; CIP, ciprofloxacin; and CEF, cefsulodin; superscript R denotes resistance against the particular drug. The data for this figure is provided in Figure 5—figure supplement 1—source data 1 . Dose-response curves data of surviving populations in the generalized experiment. The data is shown in Figure 5—figure supplement 1 and summarized in Figure 5 . Optical density values were recorded after 12 hr of incubation at 37 °C.
    Carbenicillin, supplied by Cellgro, used in various techniques. Bioz Stars score: 90/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/Cellgro
    Average 90 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    90/100 stars
      Buy from Supplier

    91
    Chem Impex International carbenicillin
    Collateral sensitivity. Validated collateral sensitivity for 38 distinct populations obtained from Barbosa et al. (2017) . Six to seven populations resistant to CIP, ( a ); STR, ( b ); GEN, ( c ); PIT, ( d ); CAR, ( e ); and CEF, ( f ), were tested for collaterals sensitivity against 2 or four drugs, indicated above each of the panels. The ancestor PA14 is shown in black and the various populations in distinct colors. Points indicate averages of 3 technical replicates per population and drug concentration. CAR, <t>carbenicillin;</t> GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; CIP, ciprofloxacin; and CEF, cefsulodin; superscript R denotes resistance against the particular drug. The data for this figure is provided in Figure 5—figure supplement 1—source data 1 . Dose-response curves data of surviving populations in the generalized experiment. The data is shown in Figure 5—figure supplement 1 and summarized in Figure 5 . Optical density values were recorded after 12 hr of incubation at 37 °C.
    Carbenicillin, supplied by Chem Impex International, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin/product/Chem Impex International
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    carbenicillin - by Bioz Stars, 2020-08
    91/100 stars
      Buy from Supplier

    86
    Duchefa carbenicillin disodium
    Effect of β-lactam antibiotics on plant regeneration of different carrot accessions from cefotaxime-, <t>carbenicillin-,</t> and timentin-containing protoplast culture media. Bars represent the standard error. D ‘Dolanka’, A ‘Amsterdamska’, K ‘Koral’. Means denoted by different letters are significantly different ( P ≤ .001).
    Carbenicillin Disodium, supplied by Duchefa, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/carbenicillin disodium/product/Duchefa
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    carbenicillin disodium - by Bioz Stars, 2020-08
    86/100 stars
      Buy from Supplier

    Image Search Results


    Genetic dominance limits the phenotypic contribution of horizontally transferred recessive alleles. (A) Pictures of a representative replicate of the conjugation assays. Overnight cultures of spots inoculated from 10-fold dilutions of conjugation mixes (10 0 to 10 −4 , from left to right) on plates selecting for transconjugants. Selection on carbenicillin reveals the actual number of transconjugants; selection on carbenicillin plus nalidixic acid or trimethoprim reveals the number of transconjugants carrying gyrA and folA alleles and expressing the resistant phenotype. (B) Antibiotic resistance level conferred by a plasmid-encoded resistant allele in the recipient bacterium (when a wild-type copy of the gene is present in the chromosome), assuming phenotypic resistance as the product of plasmid copy number and the coefficient of dominance of the allele. Experimental data are presented for gyrA D87N and folA L28R in plasmid pSEVA121.

    Journal: bioRxiv

    Article Title: Genetic dominance governs the evolution and spread of mobile genetic elements in bacteria

    doi: 10.1101/863472

    Figure Lengend Snippet: Genetic dominance limits the phenotypic contribution of horizontally transferred recessive alleles. (A) Pictures of a representative replicate of the conjugation assays. Overnight cultures of spots inoculated from 10-fold dilutions of conjugation mixes (10 0 to 10 −4 , from left to right) on plates selecting for transconjugants. Selection on carbenicillin reveals the actual number of transconjugants; selection on carbenicillin plus nalidixic acid or trimethoprim reveals the number of transconjugants carrying gyrA and folA alleles and expressing the resistant phenotype. (B) Antibiotic resistance level conferred by a plasmid-encoded resistant allele in the recipient bacterium (when a wild-type copy of the gene is present in the chromosome), assuming phenotypic resistance as the product of plasmid copy number and the coefficient of dominance of the allele. Experimental data are presented for gyrA D87N and folA L28R in plasmid pSEVA121.

    Article Snippet: Antibiotics were supplied by Sigma-Aldrich and were used at the following concentrations: carbenicillin 100 μg/ml, chloramphenicol 30 μg/ml, nalidixic acid 30 μg/ml, streptomycin 100 μg/ml, rifampicin 100 μg/ml, trimethoprim 2 μg/ml and 16 μg/ml, and tetracycline 15 μg/ml.

    Techniques: Conjugation Assay, Selection, Expressing, Plasmid Preparation

    Antibiotic resistance increases in final bacterial populations relative to the ancestor (assigned as MIC = 1) regarding the presence and type of antibiotic. ‘No‐phage’ and ‘phage’ indicate whether bacteria evolved with phages. Antibiotic types are assigned as ‘0’ for nonantibiotic treatments, ‘Carb’ for carbenicillin, ‘Gent’ for gentamicin, and ‘Trim’ for trimethoprim. Different antibiotic doses were grouped. Bars represent standard errors.

    Journal: Evolutionary Applications

    Article Title: Long‐term effects of single and combined introductions of antibiotics and bacteriophages on populations of Pseudomonas aeruginosa

    doi: 10.1111/eva.12364

    Figure Lengend Snippet: Antibiotic resistance increases in final bacterial populations relative to the ancestor (assigned as MIC = 1) regarding the presence and type of antibiotic. ‘No‐phage’ and ‘phage’ indicate whether bacteria evolved with phages. Antibiotic types are assigned as ‘0’ for nonantibiotic treatments, ‘Carb’ for carbenicillin, ‘Gent’ for gentamicin, and ‘Trim’ for trimethoprim. Different antibiotic doses were grouped. Bars represent standard errors.

    Article Snippet: We used the antibiotics carbenicillin, gentamicin, and trimethoprim (Sigma‐Aldrich, St. Louis, MO, USA), belonging to the following families (bacterial pathways targeted): β ‐lactam (inhibits cell wall synthesis); aminoglycoside (blocks protein synthesis); sulfamide (interferes with nucleic acid synthesis), respectively.

    Techniques:

    Synergistic effects of treatments through time depending on antibiotic type (Panels (A) carbenicillin; (B) gentamicin; (C) trimethoprim). Expected additive versus observed effects of combined phage‐antibiotic treatments in preventing growth in bacterial populations. Effects on growth are measured as decreases in density relative to the control. A synergistic effect was identified when the observed OD difference was significantly higher than the expected additive effect. The expected additive effect was calculated by summing the individual effects of phage and antibiotic treatments. Different antibiotic doses were grouped. Bars represent standard errors.

    Journal: Evolutionary Applications

    Article Title: Long‐term effects of single and combined introductions of antibiotics and bacteriophages on populations of Pseudomonas aeruginosa

    doi: 10.1111/eva.12364

    Figure Lengend Snippet: Synergistic effects of treatments through time depending on antibiotic type (Panels (A) carbenicillin; (B) gentamicin; (C) trimethoprim). Expected additive versus observed effects of combined phage‐antibiotic treatments in preventing growth in bacterial populations. Effects on growth are measured as decreases in density relative to the control. A synergistic effect was identified when the observed OD difference was significantly higher than the expected additive effect. The expected additive effect was calculated by summing the individual effects of phage and antibiotic treatments. Different antibiotic doses were grouped. Bars represent standard errors.

    Article Snippet: We used the antibiotics carbenicillin, gentamicin, and trimethoprim (Sigma‐Aldrich, St. Louis, MO, USA), belonging to the following families (bacterial pathways targeted): β ‐lactam (inhibits cell wall synthesis); aminoglycoside (blocks protein synthesis); sulfamide (interferes with nucleic acid synthesis), respectively.

    Techniques:

    The Dependence between Persister Formation Frequency and TolC Expression Level (A) TolC expression level in total cells of TC tag-TolC strain measured by Tetracysteine-based protein detection (FlAsH imaging, upper panel); TolC expression level in persister cells that survived antibiotic treatment (lower panel). (B) Time-lapse microscopy showing that persister cells express a high level of TolC (from Movie S3 ). The first and fifth images are the merged bright field and fluorescent images (separate bright field and fluorescent images are shown in Figure S3 D). The different media added during the experiment are indicated below (killing medium: 90% [v/v] M9 + 10% [v/v] LB + 150 μg/ml carbenicillin + 0.15% [w/v] methylcellulose; growth medium: 90% [v/v] M9 + 10% [v/v] LB + 5% [w/v] methylcellulose). The persister cell (arrow), shows a significantly higher expression of TolC, and regrows after removal of antibiotic (from Movie S3 ). (Scale bar, 3 μm; t = time in min). (C) Experimental procedure for sorting cells by expression level of persistence related genes. (D) Distribution of fluorescence intensity indicating TolC expression levels of 76RP strain. Stationary-phase cells were sorted into three groups: group A containing the total cells (100.0%), group B including the majority of total cells except for those with highest fluorescence intensity (97.8%), and group C containing a sub-population with highest fluorescence intensity (1.0%). (E) Bar plot representing cell survival rate (percentage log scale) after 4 hr carbenicillin treatment of three groups sorted by flow cytometer, revealing that the survival rate significantly increases in group C. The bars indicate mean of at least three independent experiments; error bar indicates SD. See also Figure S3 and Movie S3 .

    Journal: Molecular Cell

    Article Title: Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells

    doi: 10.1016/j.molcel.2016.03.035

    Figure Lengend Snippet: The Dependence between Persister Formation Frequency and TolC Expression Level (A) TolC expression level in total cells of TC tag-TolC strain measured by Tetracysteine-based protein detection (FlAsH imaging, upper panel); TolC expression level in persister cells that survived antibiotic treatment (lower panel). (B) Time-lapse microscopy showing that persister cells express a high level of TolC (from Movie S3 ). The first and fifth images are the merged bright field and fluorescent images (separate bright field and fluorescent images are shown in Figure S3 D). The different media added during the experiment are indicated below (killing medium: 90% [v/v] M9 + 10% [v/v] LB + 150 μg/ml carbenicillin + 0.15% [w/v] methylcellulose; growth medium: 90% [v/v] M9 + 10% [v/v] LB + 5% [w/v] methylcellulose). The persister cell (arrow), shows a significantly higher expression of TolC, and regrows after removal of antibiotic (from Movie S3 ). (Scale bar, 3 μm; t = time in min). (C) Experimental procedure for sorting cells by expression level of persistence related genes. (D) Distribution of fluorescence intensity indicating TolC expression levels of 76RP strain. Stationary-phase cells were sorted into three groups: group A containing the total cells (100.0%), group B including the majority of total cells except for those with highest fluorescence intensity (97.8%), and group C containing a sub-population with highest fluorescence intensity (1.0%). (E) Bar plot representing cell survival rate (percentage log scale) after 4 hr carbenicillin treatment of three groups sorted by flow cytometer, revealing that the survival rate significantly increases in group C. The bars indicate mean of at least three independent experiments; error bar indicates SD. See also Figure S3 and Movie S3 .

    Article Snippet: To one flask carbenicillin (150 μg/ml, Sigma) was added at a desired concentration to yield persisters.

    Techniques: Expressing, Imaging, Time-lapse Microscopy, Fluorescence, Flow Cytometry, Cytometry

    Persister Formation Frequency Positively Correlates with Efflux Gene Expression Level and Negatively Correlates with Intracellular Antibiotic Accumulation. Antibiotics Lethal Effects Were Enhanced by Addition of Efflux Inhibitors (A) Relative tolC expression level of the four strains measured by qPCR. WT (wild-type strain BW25113), OVER ( tolC overexpression strain, by pBAD:: tolC in BW25113, induced by 10 -5% arabinose), ΔtolC ( tolC knockout strain, Key::JW5503), and RESC ( tolC rescued strain, by pBAD:: tolC in ΔtolC strain, induced by 10 -6% arabinose). (B) Relative antibiotic accumulation in the four strains determined by fluorescence microscopy. (C) Persister formation frequency of the four strains determined by antibiotic susceptibility measurement. The correlation coefficiency between intracellular antibiotic accumulation level and tolC expression level is −0.776; between intracellular antibiotic accumulation level and probability of persistence is −0.989; between tolC expression level and probability of persistence is 0.848. (D–G) (D) Persister formation frequency of the four strains under antibiotic treatment of different concentrations. Antibiotics lethal effects were enhanced by efflux inhibitors; carbenicillin (E), cloxacillin (F), and nalidixic acid (G). Abbreviations: Car, carbenicillin. NA, nalidixic acid. Clo, cloxacillin. PAβN, Phenylalanine arginyl β-naphthylamide. NMP, 1-(1-Naphthylmethyl) piperazine. The bars indicate mean of at least three independent experiments; error bar indicates SD ( ∗ p value

    Journal: Molecular Cell

    Article Title: Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells

    doi: 10.1016/j.molcel.2016.03.035

    Figure Lengend Snippet: Persister Formation Frequency Positively Correlates with Efflux Gene Expression Level and Negatively Correlates with Intracellular Antibiotic Accumulation. Antibiotics Lethal Effects Were Enhanced by Addition of Efflux Inhibitors (A) Relative tolC expression level of the four strains measured by qPCR. WT (wild-type strain BW25113), OVER ( tolC overexpression strain, by pBAD:: tolC in BW25113, induced by 10 -5% arabinose), ΔtolC ( tolC knockout strain, Key::JW5503), and RESC ( tolC rescued strain, by pBAD:: tolC in ΔtolC strain, induced by 10 -6% arabinose). (B) Relative antibiotic accumulation in the four strains determined by fluorescence microscopy. (C) Persister formation frequency of the four strains determined by antibiotic susceptibility measurement. The correlation coefficiency between intracellular antibiotic accumulation level and tolC expression level is −0.776; between intracellular antibiotic accumulation level and probability of persistence is −0.989; between tolC expression level and probability of persistence is 0.848. (D–G) (D) Persister formation frequency of the four strains under antibiotic treatment of different concentrations. Antibiotics lethal effects were enhanced by efflux inhibitors; carbenicillin (E), cloxacillin (F), and nalidixic acid (G). Abbreviations: Car, carbenicillin. NA, nalidixic acid. Clo, cloxacillin. PAβN, Phenylalanine arginyl β-naphthylamide. NMP, 1-(1-Naphthylmethyl) piperazine. The bars indicate mean of at least three independent experiments; error bar indicates SD ( ∗ p value

    Article Snippet: To one flask carbenicillin (150 μg/ml, Sigma) was added at a desired concentration to yield persisters.

    Techniques: Expressing, Real-time Polymerase Chain Reaction, Over Expression, Knock-Out, Fluorescence, Microscopy

    Lower Intracellular Antibiotic Accumulation in Persister Cells of Wild-Type E. coli (A) Experimental procedure for measuring antibiotic accumulation in bacterial cells. (B and C) Histogram of antibiotic accumulation in total cells (n = 209) and persister cells (n = 86) of wild-type E. coli . (D) Time-lapse microscopy showing reduced accumulation of antibiotic in persisters (from Movie S1 ). The first five images are merged bright field and fluorescence images, revealing that antibiotic accumulation accompanies cell death (separate images are shown in Figure S1 F). The different media added during experiment is indicated below (killing medium: 90% [v/v] M9 + 10% [v/v] LB + 150 μg/ml carbenicillin + 20 μg/ml BOCILLIN + 0.15% [w/v] methylcellulose; growth medium: 90% [v/v] M9 + 10% [v/v] LB + 5% [w/v] methylcellulose) ( Maisonneuve et al., 2013 ). The persister cell (arrow) shows low antibiotic accumulation and regrows after removal of antibiotic (Scale bar, 3 μm; t = time in min). See also Figure S1 and Movie S1 .

    Journal: Molecular Cell

    Article Title: Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells

    doi: 10.1016/j.molcel.2016.03.035

    Figure Lengend Snippet: Lower Intracellular Antibiotic Accumulation in Persister Cells of Wild-Type E. coli (A) Experimental procedure for measuring antibiotic accumulation in bacterial cells. (B and C) Histogram of antibiotic accumulation in total cells (n = 209) and persister cells (n = 86) of wild-type E. coli . (D) Time-lapse microscopy showing reduced accumulation of antibiotic in persisters (from Movie S1 ). The first five images are merged bright field and fluorescence images, revealing that antibiotic accumulation accompanies cell death (separate images are shown in Figure S1 F). The different media added during experiment is indicated below (killing medium: 90% [v/v] M9 + 10% [v/v] LB + 150 μg/ml carbenicillin + 20 μg/ml BOCILLIN + 0.15% [w/v] methylcellulose; growth medium: 90% [v/v] M9 + 10% [v/v] LB + 5% [w/v] methylcellulose) ( Maisonneuve et al., 2013 ). The persister cell (arrow) shows low antibiotic accumulation and regrows after removal of antibiotic (Scale bar, 3 μm; t = time in min). See also Figure S1 and Movie S1 .

    Article Snippet: To one flask carbenicillin (150 μg/ml, Sigma) was added at a desired concentration to yield persisters.

    Techniques: Time-lapse Microscopy, Fluorescence

    System-wide Comparison of the Contribution of Persistence Genes to Bacterial Drug Tolerance (A) Heatmap showing relative transcript abundance of persistence related genes in total cells and persister cells. Scale below the heatmap indicates log2-normalized transcript abundance relative to the mean expression level (T, total cells; P, persister cells). See also Table S1 . (B) Bar plot representing cell survival rate (percentage log scale) after 4 hr carbenicillin treatment of two groups sorted by flow cytometer from fluorescently labeled tolC , soxS , uvrD , hipA , plsB , pspA , relE , and phoU strains, respectively. (C) Bar plot representing cell survival rate (in logarithm scale) after 4 hr carbenicillin treatment of knockout strains ΔtolC , ΔansA , Δcrp , ΔdinG , ΔdnaK , ΔdnaJ , ΔdksA , ΔhipA , ΔhupA , ΔhupB , Δlon , ΔplsB , ΔpspA , ΔruvA , ΔruvB , ΔuvrD , and ΔyigB . The bars indicate mean of at least three independent experiments; error bar indicates SD ( ∗ p value

    Journal: Molecular Cell

    Article Title: Enhanced Efflux Activity Facilitates Drug Tolerance in Dormant Bacterial Cells

    doi: 10.1016/j.molcel.2016.03.035

    Figure Lengend Snippet: System-wide Comparison of the Contribution of Persistence Genes to Bacterial Drug Tolerance (A) Heatmap showing relative transcript abundance of persistence related genes in total cells and persister cells. Scale below the heatmap indicates log2-normalized transcript abundance relative to the mean expression level (T, total cells; P, persister cells). See also Table S1 . (B) Bar plot representing cell survival rate (percentage log scale) after 4 hr carbenicillin treatment of two groups sorted by flow cytometer from fluorescently labeled tolC , soxS , uvrD , hipA , plsB , pspA , relE , and phoU strains, respectively. (C) Bar plot representing cell survival rate (in logarithm scale) after 4 hr carbenicillin treatment of knockout strains ΔtolC , ΔansA , Δcrp , ΔdinG , ΔdnaK , ΔdnaJ , ΔdksA , ΔhipA , ΔhupA , ΔhupB , Δlon , ΔplsB , ΔpspA , ΔruvA , ΔruvB , ΔuvrD , and ΔyigB . The bars indicate mean of at least three independent experiments; error bar indicates SD ( ∗ p value

    Article Snippet: To one flask carbenicillin (150 μg/ml, Sigma) was added at a desired concentration to yield persisters.

    Techniques: Expressing, Flow Cytometry, Cytometry, Labeling, Knock-Out

    SLC25A1 genetic defects confer susceptibility to mitochondrial proteostasis inhibitors. Oxygen consumption rate (OCR) was analyzed with Seahorse technology in wild type and two clones of SLC25A1 null cells. Cells were incubated in the absence or presence of drugs that target mitochondrial protein synthesis (A-I) or mitochondrial protein quality control (J). Carbenicillin was used as a control drug (O, n=3). All cells were treated for 48h with drugs except for panel A. A) Depicts a stress test after increasing incubation times with 2.25 µM doxycycline. Average ± SD n=8. Arrows A to C mark times of stress test drug injections: oligomycin, FCCP, and antimycin plus rotenone; respectively. B-D) and E-G) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline or minocycline treated SLC25A1 null and wild type cells. Average ± SE, n=4-6. H-J) Basal OCR of cells treated in the absence or presence of chloramphenicol, linezolid, or actinonin. Average ± SE, n=4-7. K-O) Basal OCR ratios for drug treated and untreated wild type and two SLC25A1 null cell clones (Δ and Δ1). P) Same as A, but wild type and SLC25A4 null cells were treated for 48h in the absence or presence of 2.25 µM doxycycline. Q) Basal OCR ratios for drug treated and untreated wild type and SLC25A4 null cells. R) Crystal violet cell survival assay for DU145 and ρ0 cells in the absence or presence of drug. Average ± SE, n=3. S) Same as A but with DU145 and ρ0 cells. DU145 cells treated in the presence of 2.25 µM doxycycline are shown by gray square symbols. Average ± SE, n=3. T-V) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline treated DU145 and ρ0 cells. Average ± SE, n=3. For A-J p values were calculated with Two-Factor ANOVA with Repeated Measures. For K-O and Q p values were calculated by One Way ANOVA followed by multiple comparison correction by the Bonferroni method. For R, p values were calculated One Way ANOVA followed by Fisher’s Least Significant Difference Comparison Test. In panels A-V, wild type and mutants are marked by gray and blue symbols, respectively.

    Journal: bioRxiv

    Article Title: Mitochondrial Proteostasis Requires Genes Encoded in a Neurodevelopmental Syndrome Locus that are Necessary for Synapse Function

    doi: 10.1101/2020.02.22.960971

    Figure Lengend Snippet: SLC25A1 genetic defects confer susceptibility to mitochondrial proteostasis inhibitors. Oxygen consumption rate (OCR) was analyzed with Seahorse technology in wild type and two clones of SLC25A1 null cells. Cells were incubated in the absence or presence of drugs that target mitochondrial protein synthesis (A-I) or mitochondrial protein quality control (J). Carbenicillin was used as a control drug (O, n=3). All cells were treated for 48h with drugs except for panel A. A) Depicts a stress test after increasing incubation times with 2.25 µM doxycycline. Average ± SD n=8. Arrows A to C mark times of stress test drug injections: oligomycin, FCCP, and antimycin plus rotenone; respectively. B-D) and E-G) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline or minocycline treated SLC25A1 null and wild type cells. Average ± SE, n=4-6. H-J) Basal OCR of cells treated in the absence or presence of chloramphenicol, linezolid, or actinonin. Average ± SE, n=4-7. K-O) Basal OCR ratios for drug treated and untreated wild type and two SLC25A1 null cell clones (Δ and Δ1). P) Same as A, but wild type and SLC25A4 null cells were treated for 48h in the absence or presence of 2.25 µM doxycycline. Q) Basal OCR ratios for drug treated and untreated wild type and SLC25A4 null cells. R) Crystal violet cell survival assay for DU145 and ρ0 cells in the absence or presence of drug. Average ± SE, n=3. S) Same as A but with DU145 and ρ0 cells. DU145 cells treated in the presence of 2.25 µM doxycycline are shown by gray square symbols. Average ± SE, n=3. T-V) Basal, ATP-dependent, and Maximal OCR of untreated and doxycycline treated DU145 and ρ0 cells. Average ± SE, n=3. For A-J p values were calculated with Two-Factor ANOVA with Repeated Measures. For K-O and Q p values were calculated by One Way ANOVA followed by multiple comparison correction by the Bonferroni method. For R, p values were calculated One Way ANOVA followed by Fisher’s Least Significant Difference Comparison Test. In panels A-V, wild type and mutants are marked by gray and blue symbols, respectively.

    Article Snippet: Drugs used were doxycycline (Sigma D9891), minocycline (Sigma M9511), chloramphenicol (Sigma C0378), Linezolid (Sigma PZ0014), Actinonin (Sigma A6671), Carbenicillin (Sigma C1389).

    Techniques: Clone Assay, Incubation, Clonogenic Cell Survival Assay

    Collateral sensitivity. Validated collateral sensitivity for 38 distinct populations obtained from Barbosa et al. (2017) . Six to seven populations resistant to CIP, ( a ); STR, ( b ); GEN, ( c ); PIT, ( d ); CAR, ( e ); and CEF, ( f ), were tested for collaterals sensitivity against 2 or four drugs, indicated above each of the panels. The ancestor PA14 is shown in black and the various populations in distinct colors. Points indicate averages of 3 technical replicates per population and drug concentration. CAR, carbenicillin; GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; CIP, ciprofloxacin; and CEF, cefsulodin; superscript R denotes resistance against the particular drug. The data for this figure is provided in Figure 5—figure supplement 1—source data 1 . Dose-response curves data of surviving populations in the generalized experiment. The data is shown in Figure 5—figure supplement 1 and summarized in Figure 5 . Optical density values were recorded after 12 hr of incubation at 37 °C.

    Journal: eLife

    Article Title: Evolutionary stability of collateral sensitivity to antibiotics in the model pathogen Pseudomonas aeruginosa

    doi: 10.7554/eLife.51481

    Figure Lengend Snippet: Collateral sensitivity. Validated collateral sensitivity for 38 distinct populations obtained from Barbosa et al. (2017) . Six to seven populations resistant to CIP, ( a ); STR, ( b ); GEN, ( c ); PIT, ( d ); CAR, ( e ); and CEF, ( f ), were tested for collaterals sensitivity against 2 or four drugs, indicated above each of the panels. The ancestor PA14 is shown in black and the various populations in distinct colors. Points indicate averages of 3 technical replicates per population and drug concentration. CAR, carbenicillin; GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; CIP, ciprofloxacin; and CEF, cefsulodin; superscript R denotes resistance against the particular drug. The data for this figure is provided in Figure 5—figure supplement 1—source data 1 . Dose-response curves data of surviving populations in the generalized experiment. The data is shown in Figure 5—figure supplement 1 and summarized in Figure 5 . Optical density values were recorded after 12 hr of incubation at 37 °C.

    Article Snippet: The resistant populations were previously selected for high levels of resistance against protein synthesis inhibitors from the aminoglycoside family, gentamicin (GEN; Carl Roth, Germany; Ref. HN09.1) and streptomycin (STR; Sigma-Aldrich, USA; Ref. S6501-5G), or alternatively cell-wall synthesis inhibitors from the β-lactam family, carbenicillin (CAR; Carl Roth, Germany; Ref. 6344.2) and piperacillin/tazobactam (PIT; Sigma-Aldrich, USA; Refs. P8396-1G and T2820-10MG).

    Techniques: Concentration Assay, Incubation

    Reciprocal collateral sensitivity and experimental design. ( a ) Two-step experimental evolution: resistant populations of P. aeruginosa PA14 were previously experimentally evolved ( Barbosa et al., 2017 ) with increasing concentrations of a particular drug (here labeled A), resulting in bacteria becoming hypersensitive to other drugs (here labeled B). In a second step, A-resistant clones were experimentally evolved in the presence of drug B, using four selection regimes: (i) strong dose increase of drug B in the presence of a constant high dose of drug A; (ii) mild dose increase of B in the presence of A; (iii) strong dose increases of B in the absence of A; and (iv) mild dose increase of B in the absence of A. Concentrations of B were increased using linear ramps starting at IC 50 (dashed black lines) and ending at levels above the IC 95 of the collaterally sensitive clone (mild increases, dashed orange line), or that of the PA14 wild type strain (strong increases, dashed black lines; detailed information on concentrations in Supplementary Table 1). ( b ) Validated reciprocity of collateral sensitivity for the isolated resistant clones and drug pair PIT/STR, and ( c ) CAR/GEN. Mean ±CI95, n = 8. Vertical dashed lines indicate the starting (black) and final doses of the mild (light orange) and strong drug increases (light blue). CAR, carbenicillin; GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; superscript R denotes resistance against the particular drug. The following supplementary table and source data are available for Figure 1b and c : Supplementary file 1 -Figure 1-supplementary table 1 and Figure 1—source data 1 . Mean optical density and CI95 values obtained after 12 hr of growth in minimal media and different antibiotics as reported in Figure 1 . The populations tested here include the PA14 wild type, and four resistant populations. Each value is the average of 8 technical replicates per bacterial population.

    Journal: eLife

    Article Title: Evolutionary stability of collateral sensitivity to antibiotics in the model pathogen Pseudomonas aeruginosa

    doi: 10.7554/eLife.51481

    Figure Lengend Snippet: Reciprocal collateral sensitivity and experimental design. ( a ) Two-step experimental evolution: resistant populations of P. aeruginosa PA14 were previously experimentally evolved ( Barbosa et al., 2017 ) with increasing concentrations of a particular drug (here labeled A), resulting in bacteria becoming hypersensitive to other drugs (here labeled B). In a second step, A-resistant clones were experimentally evolved in the presence of drug B, using four selection regimes: (i) strong dose increase of drug B in the presence of a constant high dose of drug A; (ii) mild dose increase of B in the presence of A; (iii) strong dose increases of B in the absence of A; and (iv) mild dose increase of B in the absence of A. Concentrations of B were increased using linear ramps starting at IC 50 (dashed black lines) and ending at levels above the IC 95 of the collaterally sensitive clone (mild increases, dashed orange line), or that of the PA14 wild type strain (strong increases, dashed black lines; detailed information on concentrations in Supplementary Table 1). ( b ) Validated reciprocity of collateral sensitivity for the isolated resistant clones and drug pair PIT/STR, and ( c ) CAR/GEN. Mean ±CI95, n = 8. Vertical dashed lines indicate the starting (black) and final doses of the mild (light orange) and strong drug increases (light blue). CAR, carbenicillin; GEN, gentamicin; STR, streptomycin; PIT, piperacillin with tazobactam; superscript R denotes resistance against the particular drug. The following supplementary table and source data are available for Figure 1b and c : Supplementary file 1 -Figure 1-supplementary table 1 and Figure 1—source data 1 . Mean optical density and CI95 values obtained after 12 hr of growth in minimal media and different antibiotics as reported in Figure 1 . The populations tested here include the PA14 wild type, and four resistant populations. Each value is the average of 8 technical replicates per bacterial population.

    Article Snippet: The resistant populations were previously selected for high levels of resistance against protein synthesis inhibitors from the aminoglycoside family, gentamicin (GEN; Carl Roth, Germany; Ref. HN09.1) and streptomycin (STR; Sigma-Aldrich, USA; Ref. S6501-5G), or alternatively cell-wall synthesis inhibitors from the β-lactam family, carbenicillin (CAR; Carl Roth, Germany; Ref. 6344.2) and piperacillin/tazobactam (PIT; Sigma-Aldrich, USA; Refs. P8396-1G and T2820-10MG).

    Techniques: Labeling, Clone Assay, Selection, Isolation

    Re-sensitization to gentamicin (GEN) upon adaptation to carbenicillin (CAR). We calculated ( a ) dose-response relationships against GEN of 15 populations adapted to strong (n = 7, light blue) and mild (n = 8, light orange) drug increases compared to the PA14 ancestor (black, bottom-right panel). Mean ±CI95, n = 3 technical replicates. In most cases, the evolved population had the same MIC as PA14. Two populations (aF7 and aG7) showed lower MICs than PA14, while two (aH8 and bH8) showed slightly higher ones. The labels within each graph correspond to the code used during experimental evolution. Data from Source Data 4 ( b ) Difference in the area under the curve (AUC) between each evaluated population and the PA14 ancestor. Scaling of the y-axis is equivalent to Figure 3d . None of the populations was significantly different from the ancestor (Wilcoxon’s test, n = 3, adjusted P values min > 0.4, and max

    Journal: eLife

    Article Title: Evolutionary stability of collateral sensitivity to antibiotics in the model pathogen Pseudomonas aeruginosa

    doi: 10.7554/eLife.51481

    Figure Lengend Snippet: Re-sensitization to gentamicin (GEN) upon adaptation to carbenicillin (CAR). We calculated ( a ) dose-response relationships against GEN of 15 populations adapted to strong (n = 7, light blue) and mild (n = 8, light orange) drug increases compared to the PA14 ancestor (black, bottom-right panel). Mean ±CI95, n = 3 technical replicates. In most cases, the evolved population had the same MIC as PA14. Two populations (aF7 and aG7) showed lower MICs than PA14, while two (aH8 and bH8) showed slightly higher ones. The labels within each graph correspond to the code used during experimental evolution. Data from Source Data 4 ( b ) Difference in the area under the curve (AUC) between each evaluated population and the PA14 ancestor. Scaling of the y-axis is equivalent to Figure 3d . None of the populations was significantly different from the ancestor (Wilcoxon’s test, n = 3, adjusted P values min > 0.4, and max

    Article Snippet: The resistant populations were previously selected for high levels of resistance against protein synthesis inhibitors from the aminoglycoside family, gentamicin (GEN; Carl Roth, Germany; Ref. HN09.1) and streptomycin (STR; Sigma-Aldrich, USA; Ref. S6501-5G), or alternatively cell-wall synthesis inhibitors from the β-lactam family, carbenicillin (CAR; Carl Roth, Germany; Ref. 6344.2) and piperacillin/tazobactam (PIT; Sigma-Aldrich, USA; Refs. P8396-1G and T2820-10MG).

    Techniques:

    Effect of β-lactam antibiotics on plant regeneration of different carrot accessions from cefotaxime-, carbenicillin-, and timentin-containing protoplast culture media. Bars represent the standard error. D ‘Dolanka’, A ‘Amsterdamska’, K ‘Koral’. Means denoted by different letters are significantly different ( P ≤ .001).

    Journal: In Vitro Cellular & Developmental Biology

    Article Title: Effect of β-lactam antibiotics on plant regeneration in carrot protoplast cultures

    doi: 10.1007/s11627-014-9626-0

    Figure Lengend Snippet: Effect of β-lactam antibiotics on plant regeneration of different carrot accessions from cefotaxime-, carbenicillin-, and timentin-containing protoplast culture media. Bars represent the standard error. D ‘Dolanka’, A ‘Amsterdamska’, K ‘Koral’. Means denoted by different letters are significantly different ( P ≤ .001).

    Article Snippet: Three types of β-lactam antibiotics were used in the experiments: cefotaxime sodium (Polfa-Tarchomin S.A., Warszawa, Poland), carbenicillin disodium (Duchefa), and timentin (ticarcillin disodium/clavulanate potassium = 1,500/100; GlaxoSmithKline, London, UK).

    Techniques: