a baumannii atcc 19606  (ATCC)


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    Name:
    Acinetobacter baumannii 2208 81 DSM 6974
    Description:

    Catalog Number:
    19606
    Price:
    None
    Applications:
    Media testingQuality control strainQuality control strain for bioMérieux VITEK and Sensititre productsFood testing
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    Structured Review

    ATCC a baumannii atcc 19606
    Overview of the complete A. <t>baumannii</t> ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

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    Images

    1) Product Images from "Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606"

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    Journal: International journal of medical microbiology : IJMM

    doi: 10.1016/j.ijmm.2020.151412

    Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).
    Figure Legend Snippet: Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Techniques Used: Plasmid Preparation

    Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.
    Figure Legend Snippet: Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Techniques Used: Sequencing

    Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.
    Figure Legend Snippet: Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Techniques Used:

    2) Product Images from "Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606"

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    Journal: International journal of medical microbiology : IJMM

    doi: 10.1016/j.ijmm.2020.151412

    Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).
    Figure Legend Snippet: Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Techniques Used: Plasmid Preparation

    Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.
    Figure Legend Snippet: Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Techniques Used: Sequencing

    Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.
    Figure Legend Snippet: Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Techniques Used:

    3) Product Images from "Polymyxin B in combination with doripenem against heteroresistant Acinetobacter baumannii: pharmacodynamics of new dosing strategies"

    Article Title: Polymyxin B in combination with doripenem against heteroresistant Acinetobacter baumannii: pharmacodynamics of new dosing strategies

    Journal: Journal of Antimicrobial Chemotherapy

    doi: 10.1093/jac/dkw293

    An HFIM was utilized to simulate various mono and combination regimens against a high inoculum of 10 9 cfu/mL of A. baumannii ATCC 19606. PMB, polymyxin B; DOR, doripenem. The total bacterial counts for the following regimens simulated: Control, growth control with no antibiotic; DOR (25 mg/L q8h) monotherapy, DOR traditional monotherapy ( fC max of 25 mg/L every 8 h); PMB (2 mg/L CI) monotherapy, PMB traditional monotherapy ( fC ss of 2 mg/L administered as a continuous infusion); PMB (5 mg/L CI) monotherapy, fC ss of 5 mg/L administered as a continuous infusion; PMB (2 mg/L CI) + DOR Burst (25 mg/L q8h for 24 h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ‘burst’ ( fC max of 25 mg/L every 8 h × 3 doses followed by no DOR thereafter); PMB (2 mg/L CI) + DOR (25 mg/L q8h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (2 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (5 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 2 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Front-Loading (5 mg/L CI on day 1 then 2 mg/L CI thereafter) + DOR (25 mg/L q8h), combination of PMB ‘front-loading’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by fC ss of 2 mg/L thereafter) and DOR ( fC max of 25 mg/L every 8 h). This figure appears in colour in the online version of JAC and in black and white in the print version of JAC .
    Figure Legend Snippet: An HFIM was utilized to simulate various mono and combination regimens against a high inoculum of 10 9 cfu/mL of A. baumannii ATCC 19606. PMB, polymyxin B; DOR, doripenem. The total bacterial counts for the following regimens simulated: Control, growth control with no antibiotic; DOR (25 mg/L q8h) monotherapy, DOR traditional monotherapy ( fC max of 25 mg/L every 8 h); PMB (2 mg/L CI) monotherapy, PMB traditional monotherapy ( fC ss of 2 mg/L administered as a continuous infusion); PMB (5 mg/L CI) monotherapy, fC ss of 5 mg/L administered as a continuous infusion; PMB (2 mg/L CI) + DOR Burst (25 mg/L q8h for 24 h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ‘burst’ ( fC max of 25 mg/L every 8 h × 3 doses followed by no DOR thereafter); PMB (2 mg/L CI) + DOR (25 mg/L q8h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (2 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (5 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 2 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Front-Loading (5 mg/L CI on day 1 then 2 mg/L CI thereafter) + DOR (25 mg/L q8h), combination of PMB ‘front-loading’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by fC ss of 2 mg/L thereafter) and DOR ( fC max of 25 mg/L every 8 h). This figure appears in colour in the online version of JAC and in black and white in the print version of JAC .

    Techniques Used:

    4) Product Images from "Polymyxin B in combination with doripenem against heteroresistant Acinetobacter baumannii: pharmacodynamics of new dosing strategies"

    Article Title: Polymyxin B in combination with doripenem against heteroresistant Acinetobacter baumannii: pharmacodynamics of new dosing strategies

    Journal: Journal of Antimicrobial Chemotherapy

    doi: 10.1093/jac/dkw293

    An HFIM was utilized to simulate various mono and combination regimens against a high inoculum of 10 9 cfu/mL of A. baumannii ATCC 19606. PMB, polymyxin B; DOR, doripenem. The total bacterial counts for the following regimens simulated: Control, growth control with no antibiotic; DOR (25 mg/L q8h) monotherapy, DOR traditional monotherapy ( fC max of 25 mg/L every 8 h); PMB (2 mg/L CI) monotherapy, PMB traditional monotherapy ( fC ss of 2 mg/L administered as a continuous infusion); PMB (5 mg/L CI) monotherapy, fC ss of 5 mg/L administered as a continuous infusion; PMB (2 mg/L CI) + DOR Burst (25 mg/L q8h for 24 h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ‘burst’ ( fC max of 25 mg/L every 8 h × 3 doses followed by no DOR thereafter); PMB (2 mg/L CI) + DOR (25 mg/L q8h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (2 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (5 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 2 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Front-Loading (5 mg/L CI on day 1 then 2 mg/L CI thereafter) + DOR (25 mg/L q8h), combination of PMB ‘front-loading’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by fC ss of 2 mg/L thereafter) and DOR ( fC max of 25 mg/L every 8 h). This figure appears in colour in the online version of JAC and in black and white in the print version of JAC .
    Figure Legend Snippet: An HFIM was utilized to simulate various mono and combination regimens against a high inoculum of 10 9 cfu/mL of A. baumannii ATCC 19606. PMB, polymyxin B; DOR, doripenem. The total bacterial counts for the following regimens simulated: Control, growth control with no antibiotic; DOR (25 mg/L q8h) monotherapy, DOR traditional monotherapy ( fC max of 25 mg/L every 8 h); PMB (2 mg/L CI) monotherapy, PMB traditional monotherapy ( fC ss of 2 mg/L administered as a continuous infusion); PMB (5 mg/L CI) monotherapy, fC ss of 5 mg/L administered as a continuous infusion; PMB (2 mg/L CI) + DOR Burst (25 mg/L q8h for 24 h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ‘burst’ ( fC max of 25 mg/L every 8 h × 3 doses followed by no DOR thereafter); PMB (2 mg/L CI) + DOR (25 mg/L q8h), combination of PMB traditional ( fC ss of 2 mg/L continuous infusion) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (2 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Burst (5 mg/L CI on day 1) + DOR (25 mg/L q8h), combination of PMB ‘burst’ ( fC ss of 2 mg/L continuous infusion for 24 h followed by no PMB thereafter) and DOR ( fC max of 25 mg/L every 8 h); PMB Front-Loading (5 mg/L CI on day 1 then 2 mg/L CI thereafter) + DOR (25 mg/L q8h), combination of PMB ‘front-loading’ ( fC ss of 5 mg/L continuous infusion for 24 h followed by fC ss of 2 mg/L thereafter) and DOR ( fC max of 25 mg/L every 8 h). This figure appears in colour in the online version of JAC and in black and white in the print version of JAC .

    Techniques Used:

    5) Product Images from "Different surface charge of colistin-susceptible and -resistant Acinetobacter baumannii cells measured with zeta potential as a function of growth phase and colistin treatment"

    Article Title: Different surface charge of colistin-susceptible and -resistant Acinetobacter baumannii cells measured with zeta potential as a function of growth phase and colistin treatment

    Journal: Journal of Antimicrobial Chemotherapy

    doi: 10.1093/jac/dkq422

    (a) The increase in mean zeta potential for a series of paired A. baumannii strains at mid-logarithmic phase following treatment with 8 mg/L colistin and PBN. (b and c) The effect of increasing concentrations of colistin and PBN, respectively, on the zeta potential of paired colistin-susceptible and -resistant A. baumannii ATCC 19606 and FADDI-AB016 at mid-logarithmic phase. Data shown represent the mean ± SD.
    Figure Legend Snippet: (a) The increase in mean zeta potential for a series of paired A. baumannii strains at mid-logarithmic phase following treatment with 8 mg/L colistin and PBN. (b and c) The effect of increasing concentrations of colistin and PBN, respectively, on the zeta potential of paired colistin-susceptible and -resistant A. baumannii ATCC 19606 and FADDI-AB016 at mid-logarithmic phase. Data shown represent the mean ± SD.

    Techniques Used:

    Zeta potential (mean ± SD) of A. baumannii ATCC 19606, and paired colistin-resistant strain ATCC 19606R, at mid-logarithmic phase as a function of KCl ionic strength at pH 5.5.
    Figure Legend Snippet: Zeta potential (mean ± SD) of A. baumannii ATCC 19606, and paired colistin-resistant strain ATCC 19606R, at mid-logarithmic phase as a function of KCl ionic strength at pH 5.5.

    Techniques Used:

    6) Product Images from "Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains"

    Article Title: Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains

    Journal: BMC Microbiology

    doi: 10.1186/s12866-020-1722-1

    Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains
    Figure Legend Snippet: Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains

    Techniques Used: Blocking Assay, Expressing

    Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively
    Figure Legend Snippet: Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively

    Techniques Used: Expressing

    Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms
    Figure Legend Snippet: Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms

    Techniques Used: Software

    a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains
    Figure Legend Snippet: a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains

    Techniques Used:

    qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630
    Figure Legend Snippet: qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630

    Techniques Used: Quantitative RT-PCR

    Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline
    Figure Legend Snippet: Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline

    Techniques Used: Transmission Assay

    7) Product Images from "Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains"

    Article Title: Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains

    Journal: BMC Microbiology

    doi: 10.1186/s12866-020-1722-1

    Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains
    Figure Legend Snippet: Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains

    Techniques Used: Blocking Assay, Expressing

    Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively
    Figure Legend Snippet: Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively

    Techniques Used: Expressing

    Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms
    Figure Legend Snippet: Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms

    Techniques Used: Software

    a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains
    Figure Legend Snippet: a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains

    Techniques Used:

    qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630
    Figure Legend Snippet: qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630

    Techniques Used: Quantitative RT-PCR

    Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline
    Figure Legend Snippet: Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline

    Techniques Used: Transmission Assay

    8) Product Images from "Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains"

    Article Title: Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains

    Journal: BMC Microbiology

    doi: 10.1186/s12866-020-1722-1

    Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains
    Figure Legend Snippet: Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains

    Techniques Used: Blocking Assay, Expressing

    Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively
    Figure Legend Snippet: Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively

    Techniques Used: Expressing

    Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms
    Figure Legend Snippet: Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms

    Techniques Used: Software

    a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains
    Figure Legend Snippet: a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains

    Techniques Used:

    qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630
    Figure Legend Snippet: qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630

    Techniques Used: Quantitative RT-PCR

    Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline
    Figure Legend Snippet: Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline

    Techniques Used: Transmission Assay

    9) Product Images from "Genes Involved in the Biosynthesis and Transport of Acinetobactin in Acinetobacter baumannii"

    Article Title: Genes Involved in the Biosynthesis and Transport of Acinetobactin in Acinetobacter baumannii

    Journal: Genomics & Informatics

    doi: 10.5808/GI.2015.13.1.2

    Genetic map of the gene cluster containing the entA ortholog. The arrows represent the coding regions of the genes identified in the genomes of Acinetobacter baumannii ATCC 19606 and ATCC 17978. The names of the genes are shown above each coding region.
    Figure Legend Snippet: Genetic map of the gene cluster containing the entA ortholog. The arrows represent the coding regions of the genes identified in the genomes of Acinetobacter baumannii ATCC 19606 and ATCC 17978. The names of the genes are shown above each coding region.

    Techniques Used:

    Physical map of siderophore gene clusters in Acinetobacter baumannii . Siderophore gene clusters in different A. baumannii strains. The gene clusters are based on the database of A. baumannii ATCC 19606, AB0057, ACICU, and AB307-294, which were retrieved separately from GenBank (accession Nos. ACQB00000000, CP001182-3, CP000863, and CP001172). Each color indicates the biological functions in the biosynthesis and transport of acinetobactin. Red arrows represent the genes that were added in the acinetobactin gene cluster.
    Figure Legend Snippet: Physical map of siderophore gene clusters in Acinetobacter baumannii . Siderophore gene clusters in different A. baumannii strains. The gene clusters are based on the database of A. baumannii ATCC 19606, AB0057, ACICU, and AB307-294, which were retrieved separately from GenBank (accession Nos. ACQB00000000, CP001182-3, CP000863, and CP001172). Each color indicates the biological functions in the biosynthesis and transport of acinetobactin. Red arrows represent the genes that were added in the acinetobactin gene cluster.

    Techniques Used:

    10) Product Images from "Role of Fibronectin in the Adhesion of Acinetobacter baumannii to Host Cells"

    Article Title: Role of Fibronectin in the Adhesion of Acinetobacter baumannii to Host Cells

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0033073

    Interaction of A. baumannii with immobilized fibronectin. ( A ) A. baumannii binding to fibronectin. ATCC 19606, 77 or 113-16 strains were incubated in BSA or in fibronectin-coated wells for 3 h at room temperature. Adherent bacteria were quantified by serial dilutions as described in materials and methods . ( B ) Inhibition of A. baumannii adherence to immobilized fibronectin by free fibronectin. ATCC 19606, 77 or 113-16 strain were incubated in fibronectin-coated wells containing increasing concentrations of free fibronectin (0, 10, 100 and 1,000 µg/mL) or BSA (1,000 µg/mL). Adherent bacteria were quantified by serial dilutions as described in materials and methods . Results were expressed as the percentage of total untreated A. baumannii adhered to immobilized fibronectin. Representative results of three independent experiments are shown and data are the means ± SEM. P
    Figure Legend Snippet: Interaction of A. baumannii with immobilized fibronectin. ( A ) A. baumannii binding to fibronectin. ATCC 19606, 77 or 113-16 strains were incubated in BSA or in fibronectin-coated wells for 3 h at room temperature. Adherent bacteria were quantified by serial dilutions as described in materials and methods . ( B ) Inhibition of A. baumannii adherence to immobilized fibronectin by free fibronectin. ATCC 19606, 77 or 113-16 strain were incubated in fibronectin-coated wells containing increasing concentrations of free fibronectin (0, 10, 100 and 1,000 µg/mL) or BSA (1,000 µg/mL). Adherent bacteria were quantified by serial dilutions as described in materials and methods . Results were expressed as the percentage of total untreated A. baumannii adhered to immobilized fibronectin. Representative results of three independent experiments are shown and data are the means ± SEM. P

    Techniques Used: Binding Assay, Incubation, Inhibition

    Involvement of A. baumannii OMPA in the adherence of A. baumannii to human lung epithelial cells. A549 cells were infected for 2 h with 10 8 cfu/mL of A. baumannii ATCC 19606, 77 or 113-16 strain previously incubated for 1 h with mouse anti-OMPA of A. baumannii (1∶1,000 and 1∶250), rabbit anti-CarO of A. baumannii (1∶25) and control mouse IgG (1∶25). Adherence assay was performed as described in materials and methods . The effect of treatment on A. baumannii adherence to A549 cells is expressed as the percentage of total untreated A. baumannii adhered to A549 cells. Representative results of three independent experiments are shown and data are the means ± SEM. P
    Figure Legend Snippet: Involvement of A. baumannii OMPA in the adherence of A. baumannii to human lung epithelial cells. A549 cells were infected for 2 h with 10 8 cfu/mL of A. baumannii ATCC 19606, 77 or 113-16 strain previously incubated for 1 h with mouse anti-OMPA of A. baumannii (1∶1,000 and 1∶250), rabbit anti-CarO of A. baumannii (1∶25) and control mouse IgG (1∶25). Adherence assay was performed as described in materials and methods . The effect of treatment on A. baumannii adherence to A549 cells is expressed as the percentage of total untreated A. baumannii adhered to A549 cells. Representative results of three independent experiments are shown and data are the means ± SEM. P

    Techniques Used: Infection, Incubation

    Immunodetection of binding of fibronectin to outer membrane proteins of A. baumannii and rOMPA. Immunodetection of FBPs outer membrane of A.baumannii ATCC 19606, 77 or 113-16 strain ( A ) and FBP rOMPA ( B ). OMPs were extracted from outer membrane and rOMPA was produced in E. coli as described in materials and methods and stained with SimplyBlue™ SafeStain (SDS-PAGE) or electrotransferred onto nitrocellulose membrane, and incubated with Fn. FBPs were probed with rabbit anti- human fibronectin and HRP-conjugated goat anti-rabbit IgG (WB). Molecular mass standards (kDa) are shown on the right. FBPs: fibronectin binding proteins, Fn: fibronectin, MW: molecular weight, WB: western blot.
    Figure Legend Snippet: Immunodetection of binding of fibronectin to outer membrane proteins of A. baumannii and rOMPA. Immunodetection of FBPs outer membrane of A.baumannii ATCC 19606, 77 or 113-16 strain ( A ) and FBP rOMPA ( B ). OMPs were extracted from outer membrane and rOMPA was produced in E. coli as described in materials and methods and stained with SimplyBlue™ SafeStain (SDS-PAGE) or electrotransferred onto nitrocellulose membrane, and incubated with Fn. FBPs were probed with rabbit anti- human fibronectin and HRP-conjugated goat anti-rabbit IgG (WB). Molecular mass standards (kDa) are shown on the right. FBPs: fibronectin binding proteins, Fn: fibronectin, MW: molecular weight, WB: western blot.

    Techniques Used: Immunodetection, Binding Assay, Produced, Staining, SDS Page, Incubation, Western Blot, Molecular Weight

    Involvement of fibronectin in the adherence of A. baumannii to human lung epithelial cells. A549 cells were pretreated with RGD (0.5 and 5 mg/mL) ( A ), rabbit anti-human fibronectin (1∶25), rat anti-human E-cadherin (1∶25) or control mouse IgG (1∶25) ( B ) and infected with 10 8 cfu/mL of A. baumannii ATCC 19606, 77 or 113-16 strain for 2 h. Adherence assay was performed as described in materials and methods . The effect of treatment on A. baumannii adherence to A549 cells is expressed as the percentage of total untreated A. baumannii adhered to A549 cells. Immunostaining for A549 cells fibronectin and A. baumannii OMPs in infected A549 cells with 10 8 cfu/mL A. baumannii ATCC 19606, 77 or 113 strains for 2 h ( C ) and 24 h ( D ) were performed and imaged by immunofluorescence microscopy. Fibronectin of A549 cells and OMPs of A. baumannii strains were detected by rabbit anti-human fibronectin and mouse anti- A. baumannii OMPs antibodies and labeled with Alexa594 and Alexa488-tagged the secondary antibodies that appeared red and green, respectively. Blue staining shows the location of A549 cells nucleus. Representative results of three independent experiments are shown and data are the means ± SEM. P
    Figure Legend Snippet: Involvement of fibronectin in the adherence of A. baumannii to human lung epithelial cells. A549 cells were pretreated with RGD (0.5 and 5 mg/mL) ( A ), rabbit anti-human fibronectin (1∶25), rat anti-human E-cadherin (1∶25) or control mouse IgG (1∶25) ( B ) and infected with 10 8 cfu/mL of A. baumannii ATCC 19606, 77 or 113-16 strain for 2 h. Adherence assay was performed as described in materials and methods . The effect of treatment on A. baumannii adherence to A549 cells is expressed as the percentage of total untreated A. baumannii adhered to A549 cells. Immunostaining for A549 cells fibronectin and A. baumannii OMPs in infected A549 cells with 10 8 cfu/mL A. baumannii ATCC 19606, 77 or 113 strains for 2 h ( C ) and 24 h ( D ) were performed and imaged by immunofluorescence microscopy. Fibronectin of A549 cells and OMPs of A. baumannii strains were detected by rabbit anti-human fibronectin and mouse anti- A. baumannii OMPs antibodies and labeled with Alexa594 and Alexa488-tagged the secondary antibodies that appeared red and green, respectively. Blue staining shows the location of A549 cells nucleus. Representative results of three independent experiments are shown and data are the means ± SEM. P

    Techniques Used: Infection, Immunostaining, Immunofluorescence, Microscopy, Labeling, Staining

    11) Product Images from "Monoclonal antibodies against the iron regulated outer membrane Proteins of Acinetobacter baumannii are bactericidal"

    Article Title: Monoclonal antibodies against the iron regulated outer membrane Proteins of Acinetobacter baumannii are bactericidal

    Journal: BMC Microbiology

    doi:

    Immunoblot of monoclonal antibodies against IROMPs of A. baumannii ATCC 19606 Immunoblot of Monoclonal antibodies with OMP of A. baumannii grown in CDM-Fe medium. Molecular Weight markers were marked in kDa. Lane A . 2G g ABIR; B . 3D 5 ABIR C . 1D 11 ABIR D . 5D 6 ABIR E . 1F 7 ABIR F . +ve control G. -ve control
    Figure Legend Snippet: Immunoblot of monoclonal antibodies against IROMPs of A. baumannii ATCC 19606 Immunoblot of Monoclonal antibodies with OMP of A. baumannii grown in CDM-Fe medium. Molecular Weight markers were marked in kDa. Lane A . 2G g ABIR; B . 3D 5 ABIR C . 1D 11 ABIR D . 5D 6 ABIR E . 1F 7 ABIR F . +ve control G. -ve control

    Techniques Used: Molecular Weight

    Effect of monoclonal antibody on iron Uptake by Acinetobacter baumannii ATCC 19606 cells using purified siderophore radiolabelled with 55 Fe. Each experiment was carried out three independent times and mean values are shown here. The bar represents the standard deviation
    Figure Legend Snippet: Effect of monoclonal antibody on iron Uptake by Acinetobacter baumannii ATCC 19606 cells using purified siderophore radiolabelled with 55 Fe. Each experiment was carried out three independent times and mean values are shown here. The bar represents the standard deviation

    Techniques Used: Purification, Standard Deviation

    SDS-PAGE profile of Outer Membrane protein of Acinetobacter baumannii ATCC 19606. Lane A is OMP form bacteria grown in presence of iron and Lane B is OMP from bacteria grown in absence of iron. Molecular weights are shown in the left most lane.
    Figure Legend Snippet: SDS-PAGE profile of Outer Membrane protein of Acinetobacter baumannii ATCC 19606. Lane A is OMP form bacteria grown in presence of iron and Lane B is OMP from bacteria grown in absence of iron. Molecular weights are shown in the left most lane.

    Techniques Used: SDS Page

    12) Product Images from "Recombinant Scorpine Produced Using SUMO Fusion Partner in Escherichia coli Has the Activities against Clinically Isolated Bacteria and Inhibits the Plasmodium falciparum Parasitemia In Vitro"

    Article Title: Recombinant Scorpine Produced Using SUMO Fusion Partner in Escherichia coli Has the Activities against Clinically Isolated Bacteria and Inhibits the Plasmodium falciparum Parasitemia In Vitro

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0103456

    Inhibition of recombinant Scorpine against the biofilm formation indices of bacteria. (A) S. aureus ATCC 29213 (B) S. aureus S (C) S. aureus R (D) A. baumannii ATCC 19606 (E) A. baumannii S (F) A. baumannii R cultured in the presence of recombinant Scorpine for 24 h. Different letters (A–C) are significantly different within treatments at p
    Figure Legend Snippet: Inhibition of recombinant Scorpine against the biofilm formation indices of bacteria. (A) S. aureus ATCC 29213 (B) S. aureus S (C) S. aureus R (D) A. baumannii ATCC 19606 (E) A. baumannii S (F) A. baumannii R cultured in the presence of recombinant Scorpine for 24 h. Different letters (A–C) are significantly different within treatments at p

    Techniques Used: Inhibition, Recombinant, Cell Culture

    Inhibition of recombinant Scorpine against the growth of planktonic bacteria. (A) S. aureus ATCC 29213 (B) S. aureus S (C) S. aureus R (D) A. baumannii ATCC 19606 (E) A. baumannii S (F) A. baumannii R cultured in the presence of recombinant Scorpine for 24 h. Different letters (A–C) are significantly different within treatments at p
    Figure Legend Snippet: Inhibition of recombinant Scorpine against the growth of planktonic bacteria. (A) S. aureus ATCC 29213 (B) S. aureus S (C) S. aureus R (D) A. baumannii ATCC 19606 (E) A. baumannii S (F) A. baumannii R cultured in the presence of recombinant Scorpine for 24 h. Different letters (A–C) are significantly different within treatments at p

    Techniques Used: Inhibition, Recombinant, Cell Culture

    13) Product Images from "Immunoprotective Efficacy of Acinetobacter baumannii Outer Membrane Protein, FilF, Predicted In silico as a Potential Vaccine Candidate"

    Article Title: Immunoprotective Efficacy of Acinetobacter baumannii Outer Membrane Protein, FilF, Predicted In silico as a Potential Vaccine Candidate

    Journal: Frontiers in Microbiology

    doi: 10.3389/fmicb.2016.00158

    Cytokine levels in the sera . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21. The mice were intra-tracheally challenged with 10 8 CFU A. baumannii ATCC 19606 at day 29, and sacrificed at 12 and 24 h post-challenge. The detection limit for all cytokines and chemokines is
    Figure Legend Snippet: Cytokine levels in the sera . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21. The mice were intra-tracheally challenged with 10 8 CFU A. baumannii ATCC 19606 at day 29, and sacrificed at 12 and 24 h post-challenge. The detection limit for all cytokines and chemokines is

    Techniques Used: Mouse Assay, Immunofluorescence

    Bacterial burden in lungs . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21. The mice were intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. Immunization with FilF reduced the bacterial burden by 2 and 4 log cycles in the lungs of pneumonia model mice sacrificed 12 and 24 h post infection, respectively. The data are presented as mean ± SD ( n = 6). p -value was determined by the one way analysis of variance (ANOVA). *** p
    Figure Legend Snippet: Bacterial burden in lungs . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21. The mice were intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. Immunization with FilF reduced the bacterial burden by 2 and 4 log cycles in the lungs of pneumonia model mice sacrificed 12 and 24 h post infection, respectively. The data are presented as mean ± SD ( n = 6). p -value was determined by the one way analysis of variance (ANOVA). *** p

    Techniques Used: Mouse Assay, Immunofluorescence, Infection

    Lung histopathology . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21, and intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. The mice were sacrificed at 12 and 24 h post-challenge and lungs were collected for histopathology. (A) The lung from an unimmunized uninfected mouse showing normal histological characters. (B) Unimmunized infected mouse lung showing increased inflammatory cell infiltration in the perivascular and peribronchial areas, and within the airway lumen (arrows) 12 h postinfection. (C) The lung from an immunized infected mouse showing mild inflammatory cell infiltration in the perivascular and peribronchial areas (arrows) 12 h postinfection. (D) The lung from an immunized infected mouse showing significantly reduced infiltration of inflammatory cells 24 h postinfection. H E, Magnification 100X.
    Figure Legend Snippet: Lung histopathology . Groups of female Balb/c mice ( n = 6) were immunized subcutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21, and intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. The mice were sacrificed at 12 and 24 h post-challenge and lungs were collected for histopathology. (A) The lung from an unimmunized uninfected mouse showing normal histological characters. (B) Unimmunized infected mouse lung showing increased inflammatory cell infiltration in the perivascular and peribronchial areas, and within the airway lumen (arrows) 12 h postinfection. (C) The lung from an immunized infected mouse showing mild inflammatory cell infiltration in the perivascular and peribronchial areas (arrows) 12 h postinfection. (D) The lung from an immunized infected mouse showing significantly reduced infiltration of inflammatory cells 24 h postinfection. H E, Magnification 100X.

    Techniques Used: Histopathology, Mouse Assay, Immunofluorescence, Infection

    Survival rate of mice . Groups of female Balb/c mice ( n = 10) were immunized sub-cutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21, and intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. The survival rates of mice were recorded for seven days.
    Figure Legend Snippet: Survival rate of mice . Groups of female Balb/c mice ( n = 10) were immunized sub-cutaneously with 20 μg FilF formulated with CFA/IFA adjuvant on day 1, 14, and 21, and intra-tracheally challenged with 10 8 CFU of A. baumannii ATCC 19606 at day 29. The survival rates of mice were recorded for seven days.

    Techniques Used: Mouse Assay, Immunofluorescence

    14) Product Images from "Untargeted metabolomics analysis reveals key pathways responsible for the synergistic killing of colistin and doripenem combination against Acinetobacter baumannii"

    Article Title: Untargeted metabolomics analysis reveals key pathways responsible for the synergistic killing of colistin and doripenem combination against Acinetobacter baumannii

    Journal: Scientific Reports

    doi: 10.1038/srep45527

    Perturbations of bacterial lipids. ( A ) Significantly perturbed lipids in A. baumannii ATCC 19606 following treatment with colistin (Col, white), doripenem (Dor, grey) and the combination (Col-Dor, black) for (i) 15 min, (ii) 1 hr, and (iii) 4 hr. Lipid names are putatively assigned based on accurate mass. ( B ) Depletion of (i) sn -glycero-3-phosphoethanolamine, and (ii) sn -glycero-3-phosphate after treatment with colistin, doripenem, and the combination across all three time points. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).
    Figure Legend Snippet: Perturbations of bacterial lipids. ( A ) Significantly perturbed lipids in A. baumannii ATCC 19606 following treatment with colistin (Col, white), doripenem (Dor, grey) and the combination (Col-Dor, black) for (i) 15 min, (ii) 1 hr, and (iii) 4 hr. Lipid names are putatively assigned based on accurate mass. ( B ) Depletion of (i) sn -glycero-3-phosphoethanolamine, and (ii) sn -glycero-3-phosphate after treatment with colistin, doripenem, and the combination across all three time points. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).

    Techniques Used:

    Central metabolic changes in the pentose phosphate pathway (PPP). Antibiotic treatment of A. baumannii ATCC 19606 significantly decreased the levels of three PPP metabolites (D-sedoheptulose 7-phosphate, D-erythrose 4-phosphate, and D-ribose 5-phosphate) that are essential anabolic precursors of related pathways. The combined colistin/doripenem significantly decreased the levels of the three precursor metabolites at all the time points. Additionally, 2-deoxy-D-ribose 5-phosphate significantly decreased followed by the combination at 1 hr and 4 hr. In the pathway flow chart (adapted from biocyc.org with reference to E. coli K-12), blue boxes indicate the metabolites that were significantly decreased and red boxes indicate the metabolites that were not significantly changed. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).
    Figure Legend Snippet: Central metabolic changes in the pentose phosphate pathway (PPP). Antibiotic treatment of A. baumannii ATCC 19606 significantly decreased the levels of three PPP metabolites (D-sedoheptulose 7-phosphate, D-erythrose 4-phosphate, and D-ribose 5-phosphate) that are essential anabolic precursors of related pathways. The combined colistin/doripenem significantly decreased the levels of the three precursor metabolites at all the time points. Additionally, 2-deoxy-D-ribose 5-phosphate significantly decreased followed by the combination at 1 hr and 4 hr. In the pathway flow chart (adapted from biocyc.org with reference to E. coli K-12), blue boxes indicate the metabolites that were significantly decreased and red boxes indicate the metabolites that were not significantly changed. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).

    Techniques Used: Flow Cytometry

    Depletion of energy and nucleotide metabolite levels. ( A ) Decreased levels of key energy-associated metabolites, ATP, NAD + and NADP + induced by colistin, doripenem, and the combination in A. baumannii ATCC 19606. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA). ( B ) Heatmap profile of relative abundance of significantly perturbed nucleotides at (i) 15 min, (ii) 1 hr, and (iii) 4 hr after treatment with colistin (Col), doripenem (Dor) and the combination (Col-Dor) (n = 4). Antibiotics decreased the levels of nucleotides, both purines and pyrimidines, in A. baumannii ATCC 19606.
    Figure Legend Snippet: Depletion of energy and nucleotide metabolite levels. ( A ) Decreased levels of key energy-associated metabolites, ATP, NAD + and NADP + induced by colistin, doripenem, and the combination in A. baumannii ATCC 19606. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA). ( B ) Heatmap profile of relative abundance of significantly perturbed nucleotides at (i) 15 min, (ii) 1 hr, and (iii) 4 hr after treatment with colistin (Col), doripenem (Dor) and the combination (Col-Dor) (n = 4). Antibiotics decreased the levels of nucleotides, both purines and pyrimidines, in A. baumannii ATCC 19606.

    Techniques Used:

    Depletion of amino sugar metabolites for peptidoglycan and lipopolysaccharide biosynthesis. ( A ) Significant decrease in the levels of two amino sugar metabolites at 1 hr and 4 hr by colistin, doripenem and the combination, and perturbation of the cell envelope biosynthesis (peptidoglycan and lipopolysaccharide biosynthesis) in A. baumannii ATCC 19606. UDP- N -acetyl-D-glucosamine (UDP-GlcNAc) is a key precursor metabolite for LPS and peptidoglycan biosynthesis. ( B ) Levels of two key metabolites of peptidoglycan biosynthesis significantly decreased after treatment with doripenem alone at 4 hr. The combination of colistin and doripenem also significantly decreased UDP- N -acetylmuramoyl-L-alanyl-D-glutamyl-meso-2,6-diaminopimeloyl-D-alanyl-D-alanine ( > 2.0-log 2 -fold) at 4 hr. The blue boxes in the flow charts indicate the metabolites that were significantly decreased. The red boxes indicate the metabolites that were not significantly changed. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).
    Figure Legend Snippet: Depletion of amino sugar metabolites for peptidoglycan and lipopolysaccharide biosynthesis. ( A ) Significant decrease in the levels of two amino sugar metabolites at 1 hr and 4 hr by colistin, doripenem and the combination, and perturbation of the cell envelope biosynthesis (peptidoglycan and lipopolysaccharide biosynthesis) in A. baumannii ATCC 19606. UDP- N -acetyl-D-glucosamine (UDP-GlcNAc) is a key precursor metabolite for LPS and peptidoglycan biosynthesis. ( B ) Levels of two key metabolites of peptidoglycan biosynthesis significantly decreased after treatment with doripenem alone at 4 hr. The combination of colistin and doripenem also significantly decreased UDP- N -acetylmuramoyl-L-alanyl-D-glutamyl-meso-2,6-diaminopimeloyl-D-alanyl-D-alanine ( > 2.0-log 2 -fold) at 4 hr. The blue boxes in the flow charts indicate the metabolites that were significantly decreased. The red boxes indicate the metabolites that were not significantly changed. Box plots indicate upper and lower quartiles (top and bottom of box); median (line within box); and the spread of data that are not outliers (whiskers). *≥1.5-log 2 -fold, p ≤ 0.05, FDR ≤ 0.1 (one-way ANOVA).

    Techniques Used: Flow Cytometry

    15) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

    Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

    Journal: mSphere

    doi: 10.1128/mSphere.00199-17

    Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.
    Figure Legend Snippet: Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.
    Figure Legend Snippet: Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
    Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

    Techniques Used: Expressing, Incubation, Fluorescence, Software

    Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.
    Figure Legend Snippet: Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.

    Techniques Used: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy

    16) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

    Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

    Journal: mSphere

    doi: 10.1128/mSphere.00199-17

    Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.
    Figure Legend Snippet: Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.
    Figure Legend Snippet: Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
    Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

    Techniques Used: Expressing, Incubation, Fluorescence, Software

    Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.
    Figure Legend Snippet: Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.

    Techniques Used: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy

    17) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

    Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

    Journal: mSphere

    doi: 10.1128/mSphere.00199-17

    Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.
    Figure Legend Snippet: Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.
    Figure Legend Snippet: Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
    Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

    Techniques Used: Expressing, Incubation, Fluorescence, Software

    Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.
    Figure Legend Snippet: Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.

    Techniques Used: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy

    18) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

    Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

    Journal: mSphere

    doi: 10.1128/mSphere.00199-17

    Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.
    Figure Legend Snippet: Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.
    Figure Legend Snippet: Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
    Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

    Techniques Used: Expressing, Incubation, Fluorescence, Software

    Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.
    Figure Legend Snippet: Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.

    Techniques Used: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy

    19) Product Images from "iCN718, an Updated and Improved Genome-Scale Metabolic Network Reconstruction of Acinetobacter baumannii AYE"

    Article Title: iCN718, an Updated and Improved Genome-Scale Metabolic Network Reconstruction of Acinetobacter baumannii AYE

    Journal: Frontiers in Genetics

    doi: 10.3389/fgene.2018.00121

    Workflow of the reconstruction process. The starting reconstruction, AbyMBEL891, was cross referenced against a draft model generated utilizing ModelSEED ( Henry et al., 2010 ). Next, the reconstruction was standardized using various databases mapped to standard BIGGs IDs. This process was followed by manual curation based on current literature on the organism, aided by the use of ESCHER to visualize pathways throughout the process. Finally, the model was evaluated against experimental datasets and compared to iLP844 a model of Acinetobacter baumannii ATCC 19606 to further improve the reconstruction. The model was iteratively evaluated against gene essentiality and phenotypic datasets to improve the reconstruction accuracy.
    Figure Legend Snippet: Workflow of the reconstruction process. The starting reconstruction, AbyMBEL891, was cross referenced against a draft model generated utilizing ModelSEED ( Henry et al., 2010 ). Next, the reconstruction was standardized using various databases mapped to standard BIGGs IDs. This process was followed by manual curation based on current literature on the organism, aided by the use of ESCHER to visualize pathways throughout the process. Finally, the model was evaluated against experimental datasets and compared to iLP844 a model of Acinetobacter baumannii ATCC 19606 to further improve the reconstruction. The model was iteratively evaluated against gene essentiality and phenotypic datasets to improve the reconstruction accuracy.

    Techniques Used: Generated

    20) Product Images from "The Acinetobacter trimeric autotransporter adhesin Ata controls key virulence traits of Acinetobacter baumannii"

    Article Title: The Acinetobacter trimeric autotransporter adhesin Ata controls key virulence traits of Acinetobacter baumannii

    Journal: Virulence

    doi: 10.1080/21505594.2018.1558693

    The phylogenetic distribution of ata . (a) The tree displays the prevalence of ata in individual Acinetobacter spp.. Branch labels denote the number of genomes harboring ata vs. the total number of analyzed genomes in the corresponding clade. Species outside the Acinetobacter calcoaceticus-baumannii complex are collapsed into a single taxon named “Other Acinetobacter” . The tree topology follows Poppel et al. [ 62 ]. (b) Phylogenetic profile of the A. baumannii ATCC 19606 gene cluster encoding the following five proteins: 23S rRNA methyltransferase – Lon protease – Ata – TpgA – Cyclo-ligase. Blue dots indicate the presence of a gene’s ortholog in the respective taxon. Dot sizes are proportional to the fraction of genomes subsumed in each taxon harboring an ortholog. The presence/absence information is given per strain in the case of A. baumannii and is summarized on the species level for the other species (see Figure 5(a) for the number of analyzed genomes). The profiles of the two reference strains are shaded in grey. Further 16 profiles of A. baumannii strains exemplifying the variation within this species are depicted in the blue shaded area together with profiles of the two further species in the NSB clade. The profiles for the CDP clade are shaded in red. Ata and TpgA orthologs are absent in almost all analyzed genomes in this clade, while the remaining three genes are consistently present.
    Figure Legend Snippet: The phylogenetic distribution of ata . (a) The tree displays the prevalence of ata in individual Acinetobacter spp.. Branch labels denote the number of genomes harboring ata vs. the total number of analyzed genomes in the corresponding clade. Species outside the Acinetobacter calcoaceticus-baumannii complex are collapsed into a single taxon named “Other Acinetobacter” . The tree topology follows Poppel et al. [ 62 ]. (b) Phylogenetic profile of the A. baumannii ATCC 19606 gene cluster encoding the following five proteins: 23S rRNA methyltransferase – Lon protease – Ata – TpgA – Cyclo-ligase. Blue dots indicate the presence of a gene’s ortholog in the respective taxon. Dot sizes are proportional to the fraction of genomes subsumed in each taxon harboring an ortholog. The presence/absence information is given per strain in the case of A. baumannii and is summarized on the species level for the other species (see Figure 5(a) for the number of analyzed genomes). The profiles of the two reference strains are shaded in grey. Further 16 profiles of A. baumannii strains exemplifying the variation within this species are depicted in the blue shaded area together with profiles of the two further species in the NSB clade. The profiles for the CDP clade are shaded in red. Ata and TpgA orthologs are absent in almost all analyzed genomes in this clade, while the remaining three genes are consistently present.

    Techniques Used:

    21) Product Images from "Biological Cost of Different Mechanisms of Colistin Resistance and Their Impact on Virulence in Acinetobacter baumannii"

    Article Title: Biological Cost of Different Mechanisms of Colistin Resistance and Their Impact on Virulence in Acinetobacter baumannii

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.01597-13

    Abilities of different A. baumannii strains to cause cell death of A549 alveolar cells. (A) Fluorescence microscopy images of human alveolar A549 cells infected with each of the A. baumannii strains and stained with the LIVE/DEAD Cellstain double-staining kit. Healthy cells with intact membranes are stained green, and dead cells with permeabilized membranes are stained red. A549 cells were incubated with the A. baumannii strains ATCC 19606 WT, AL1851 Δ lpxA , Al1852 Δ lpxD , AL1842 Δ lpxC , and ATCC 19606 pmrB for 20 h or left uninfected. (B) Quantification of A549 cell death caused by A. baumannii ATCC 19606 WT and AL1851 Δ lpxA , AL1852 Δ lpxD , and AL1842 Δ lpxC mutants and the pmrB mutant. The results of 6 independent experiments are shown as means and SD. *, P
    Figure Legend Snippet: Abilities of different A. baumannii strains to cause cell death of A549 alveolar cells. (A) Fluorescence microscopy images of human alveolar A549 cells infected with each of the A. baumannii strains and stained with the LIVE/DEAD Cellstain double-staining kit. Healthy cells with intact membranes are stained green, and dead cells with permeabilized membranes are stained red. A549 cells were incubated with the A. baumannii strains ATCC 19606 WT, AL1851 Δ lpxA , Al1852 Δ lpxD , AL1842 Δ lpxC , and ATCC 19606 pmrB for 20 h or left uninfected. (B) Quantification of A549 cell death caused by A. baumannii ATCC 19606 WT and AL1851 Δ lpxA , AL1852 Δ lpxD , and AL1842 Δ lpxC mutants and the pmrB mutant. The results of 6 independent experiments are shown as means and SD. *, P

    Techniques Used: Fluorescence, Microscopy, Infection, Staining, Double Staining, Incubation, Mutagenesis

    22) Product Images from "In Vivo Efficacy of Glycopeptide-Colistin Combination Therapies in a Galleria mellonella Model of Acinetobacter baumannii Infection ▿"

    Article Title: In Vivo Efficacy of Glycopeptide-Colistin Combination Therapies in a Galleria mellonella Model of Acinetobacter baumannii Infection ▿

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00230-11

    Survival curves for A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL), gentamicin (GENT), teicoplanin (TEIC), or PBS alone. Curves represent a single experiment performed using 16 insects.
    Figure Legend Snippet: Survival curves for A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL), gentamicin (GENT), teicoplanin (TEIC), or PBS alone. Curves represent a single experiment performed using 16 insects.

    Techniques Used:

    Survival of A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL) in combination with teicoplanin (TEIC) or vancomycin (VANC). For ATCC 19606, P
    Figure Legend Snippet: Survival of A. baumannii ATCC 19606 (A) and AB210 (B) following treatment with colistin (COL) in combination with teicoplanin (TEIC) or vancomycin (VANC). For ATCC 19606, P

    Techniques Used:

    23) Product Images from "Simple Method for Markerless Gene Deletion in Multidrug-Resistant Acinetobacter baumannii"

    Article Title: Simple Method for Markerless Gene Deletion in Multidrug-Resistant Acinetobacter baumannii

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.03975-14

    Construction of basD and bauA mutants. (A) In PCR-1, the downstream and upstream regions of basD and bauA were amplified by PCR using the genomic DNA of A. baumannii ATCC 19606. nptI , conferring kanamycin resistance, was also amplified by PCR using pUC4K as a template. In PCR-2, the DNA fragments with the target genes deleted were combined with nptI by overlap extension PCR. UP, upstream region of the target gene to be deleted; DW, downstream region of the target gene to be deleted. (B) PCR analysis of the wild-type and mutant strains generated by allelic exchange. Molecular size markers (1-kb DNA ladder; Kapa) are indicated. (C) The growth of the strains on LB medium without and with 0.2 mM DIP was measured after 10 h. Error bars represent SEM. Means and SEM were calculated from three independent experiments. Asterisks indicate significant differences ( P
    Figure Legend Snippet: Construction of basD and bauA mutants. (A) In PCR-1, the downstream and upstream regions of basD and bauA were amplified by PCR using the genomic DNA of A. baumannii ATCC 19606. nptI , conferring kanamycin resistance, was also amplified by PCR using pUC4K as a template. In PCR-2, the DNA fragments with the target genes deleted were combined with nptI by overlap extension PCR. UP, upstream region of the target gene to be deleted; DW, downstream region of the target gene to be deleted. (B) PCR analysis of the wild-type and mutant strains generated by allelic exchange. Molecular size markers (1-kb DNA ladder; Kapa) are indicated. (C) The growth of the strains on LB medium without and with 0.2 mM DIP was measured after 10 h. Error bars represent SEM. Means and SEM were calculated from three independent experiments. Asterisks indicate significant differences ( P

    Techniques Used: Polymerase Chain Reaction, Amplification, Mutagenesis, Generated

    Gene deletion in A. baumannii strains other than ATCC 19606. (A) The susceptibilities of A. baumannii strains ATCC 17978 and 1656-2 to increasing amounts of antibiotics were explored by measuring their growth after 12 h on LB medium containing different concentrations of antibiotics. Asterisks indicate significant differences ( P
    Figure Legend Snippet: Gene deletion in A. baumannii strains other than ATCC 19606. (A) The susceptibilities of A. baumannii strains ATCC 17978 and 1656-2 to increasing amounts of antibiotics were explored by measuring their growth after 12 h on LB medium containing different concentrations of antibiotics. Asterisks indicate significant differences ( P

    Techniques Used:

    24) Product Images from "Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606"

    Article Title: Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00692-13

    Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione
    Figure Legend Snippet: Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione

    Techniques Used: Concentration Assay

    Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used
    Figure Legend Snippet: Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used

    Techniques Used: Mutagenesis

    Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular
    Figure Legend Snippet: Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular

    Techniques Used:

    25) Product Images from "Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606"

    Article Title: Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00692-13

    Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione
    Figure Legend Snippet: Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione

    Techniques Used: Concentration Assay

    Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used
    Figure Legend Snippet: Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used

    Techniques Used: Mutagenesis

    Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular
    Figure Legend Snippet: Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular

    Techniques Used:

    26) Product Images from "Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606"

    Article Title: Homeostasis of Glutathione Is Associated with Polyamine-Mediated ?-Lactam Susceptibility in Acinetobacter baumannii ATCC 19606

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00692-13

    Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione
    Figure Legend Snippet: Glutathione synthesis in A. baumannii ATCC 19606. Cell cultures collected at each optical density (OD 600 ) were used to measure the glutathione concentration as described in Materials and Methods. Open bars indicate extracellular levels of glutathione

    Techniques Used: Concentration Assay

    Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used
    Figure Legend Snippet: Intracellular glutathione concentrations in A. baumannii ATCC 19606 and its mutant strain (a homolog of AB_AB1837::Km of A. baumannii AB0057). Cell pellets of the mutant strain and the parental strain collected at each optical density (OD 600 ) were used

    Techniques Used: Mutagenesis

    Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular
    Figure Legend Snippet: Polyamine effects on glutathione transport in A. baumannii ATCC 19606. Cells grown with or without polyamines (1 mM spermine or 10 mM spermidine) were collected at an OD 600 of 1.0. Cell supernatants and cell pellets were used to measure levels of extracellular

    Techniques Used:

    27) Product Images from "Characterization of an Acinetobacter baumanniilptD Deletion Strain: Permeability Defects and Response to Inhibition of Lipopolysaccharide and Fatty Acid Biosynthesis"

    Article Title: Characterization of an Acinetobacter baumanniilptD Deletion Strain: Permeability Defects and Response to Inhibition of Lipopolysaccharide and Fatty Acid Biosynthesis

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00639-15

    (A) Treatment of the lptD ::Km r mutant with 0.125 μg/ml cerulenin partially restores OM integrity, as indicated by a reduction in uptake of the hydrophobic fluorescent probe NPN. The A. baumannii ATCC 19606 parent strain was treated with 1 μg/ml
    Figure Legend Snippet: (A) Treatment of the lptD ::Km r mutant with 0.125 μg/ml cerulenin partially restores OM integrity, as indicated by a reduction in uptake of the hydrophobic fluorescent probe NPN. The A. baumannii ATCC 19606 parent strain was treated with 1 μg/ml

    Techniques Used: Mutagenesis

    LCMS-MRM quantification of lipid A precursors in A. baumannii mutants. Data are representative of results from three independent experiments performed in duplicate. The P value is > 0.05 between the ATCC 19606 parent and the lptD ::Km r mutant for
    Figure Legend Snippet: LCMS-MRM quantification of lipid A precursors in A. baumannii mutants. Data are representative of results from three independent experiments performed in duplicate. The P value is > 0.05 between the ATCC 19606 parent and the lptD ::Km r mutant for

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mutagenesis

    A defect in LPS assembly ( lptD ::Km r ) and complete loss of LPS ( lpxC ::Km r ) cause similarly impaired in vitro growth compared to growth of the A. baumannii ATCC 19606 parent. (A) Data representing the means of results from three independent replicates and
    Figure Legend Snippet: A defect in LPS assembly ( lptD ::Km r ) and complete loss of LPS ( lpxC ::Km r ) cause similarly impaired in vitro growth compared to growth of the A. baumannii ATCC 19606 parent. (A) Data representing the means of results from three independent replicates and

    Techniques Used: In Vitro

    (A) Loss of lptD causes a reduction in the amount of cell-associated LPS. Lane 1, E. coli LPS standard; lane 2, A. baumannii ATCC 19606 parent strain; lane 3, A. baumannii lpxC ::Km r strain; lane 4, A. baumannii lptD ::Km r strain. The LPS gel results are
    Figure Legend Snippet: (A) Loss of lptD causes a reduction in the amount of cell-associated LPS. Lane 1, E. coli LPS standard; lane 2, A. baumannii ATCC 19606 parent strain; lane 3, A. baumannii lpxC ::Km r strain; lane 4, A. baumannii lptD ::Km r strain. The LPS gel results are

    Techniques Used:

    Transmission electron microscopy images of A. baumannii ATCC 19606 and the lptD ::Km r mutant. Deletion of lptD did not produce obvious internal accumulations of membrane-like materials.
    Figure Legend Snippet: Transmission electron microscopy images of A. baumannii ATCC 19606 and the lptD ::Km r mutant. Deletion of lptD did not produce obvious internal accumulations of membrane-like materials.

    Techniques Used: Transmission Assay, Electron Microscopy, Mutagenesis

    28) Product Images from "Characterization of an Acinetobacter baumanniilptD Deletion Strain: Permeability Defects and Response to Inhibition of Lipopolysaccharide and Fatty Acid Biosynthesis"

    Article Title: Characterization of an Acinetobacter baumanniilptD Deletion Strain: Permeability Defects and Response to Inhibition of Lipopolysaccharide and Fatty Acid Biosynthesis

    Journal: Journal of Bacteriology

    doi: 10.1128/JB.00639-15

    (A) Treatment of the lptD ::Km r mutant with 0.125 μg/ml cerulenin partially restores OM integrity, as indicated by a reduction in uptake of the hydrophobic fluorescent probe NPN. The A. baumannii ATCC 19606 parent strain was treated with 1 μg/ml
    Figure Legend Snippet: (A) Treatment of the lptD ::Km r mutant with 0.125 μg/ml cerulenin partially restores OM integrity, as indicated by a reduction in uptake of the hydrophobic fluorescent probe NPN. The A. baumannii ATCC 19606 parent strain was treated with 1 μg/ml

    Techniques Used: Mutagenesis

    LCMS-MRM quantification of lipid A precursors in A. baumannii mutants. Data are representative of results from three independent experiments performed in duplicate. The P value is > 0.05 between the ATCC 19606 parent and the lptD ::Km r mutant for
    Figure Legend Snippet: LCMS-MRM quantification of lipid A precursors in A. baumannii mutants. Data are representative of results from three independent experiments performed in duplicate. The P value is > 0.05 between the ATCC 19606 parent and the lptD ::Km r mutant for

    Techniques Used: Liquid Chromatography with Mass Spectroscopy, Mutagenesis

    A defect in LPS assembly ( lptD ::Km r ) and complete loss of LPS ( lpxC ::Km r ) cause similarly impaired in vitro growth compared to growth of the A. baumannii ATCC 19606 parent. (A) Data representing the means of results from three independent replicates and
    Figure Legend Snippet: A defect in LPS assembly ( lptD ::Km r ) and complete loss of LPS ( lpxC ::Km r ) cause similarly impaired in vitro growth compared to growth of the A. baumannii ATCC 19606 parent. (A) Data representing the means of results from three independent replicates and

    Techniques Used: In Vitro

    (A) Loss of lptD causes a reduction in the amount of cell-associated LPS. Lane 1, E. coli LPS standard; lane 2, A. baumannii ATCC 19606 parent strain; lane 3, A. baumannii lpxC ::Km r strain; lane 4, A. baumannii lptD ::Km r strain. The LPS gel results are
    Figure Legend Snippet: (A) Loss of lptD causes a reduction in the amount of cell-associated LPS. Lane 1, E. coli LPS standard; lane 2, A. baumannii ATCC 19606 parent strain; lane 3, A. baumannii lpxC ::Km r strain; lane 4, A. baumannii lptD ::Km r strain. The LPS gel results are

    Techniques Used:

    Transmission electron microscopy images of A. baumannii ATCC 19606 and the lptD ::Km r mutant. Deletion of lptD did not produce obvious internal accumulations of membrane-like materials.
    Figure Legend Snippet: Transmission electron microscopy images of A. baumannii ATCC 19606 and the lptD ::Km r mutant. Deletion of lptD did not produce obvious internal accumulations of membrane-like materials.

    Techniques Used: Transmission Assay, Electron Microscopy, Mutagenesis

    29) Product Images from "Inhibition of LpxC Increases Antibiotic Susceptibility in Acinetobacter baumannii"

    Article Title: Inhibition of LpxC Increases Antibiotic Susceptibility in Acinetobacter baumannii

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00407-16

    (A) Lipid A detection after silver staining. Lane 1: ATCC 19606; lane 2: ATCC 19606 treated with 32 mg/liter of PF-5081090; lane 3: IB010. (B) Cell permeability of A. baumannii strains treated (white bars) with 32 mg/liter of PF-5081090 or left untreated
    Figure Legend Snippet: (A) Lipid A detection after silver staining. Lane 1: ATCC 19606; lane 2: ATCC 19606 treated with 32 mg/liter of PF-5081090; lane 3: IB010. (B) Cell permeability of A. baumannii strains treated (white bars) with 32 mg/liter of PF-5081090 or left untreated

    Techniques Used: Silver Staining, Permeability

    30) Product Images from "Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression"

    Article Title: Subinhibitory Concentrations of Trimethoprim and Sulfamethoxazole Prevent Biofilm Formation by Acinetobacter baumannii through Inhibition of Csu Pilus Expression

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00778-17

    Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.
    Figure Legend Snippet: Requirement for Csu pili for biofilm formation in A. baumannii ATCC 17978. (A) Representative crystal violet assay image of A. baumannii strains on polystyrene surfaces. The biofilms were stained with crystal violet and dissolved in 10% acetic acid solution. A. baumannii ATCC 17978 wild-type and complemented strains, but not the csuD mutant strain, formed biofilms. The endogenous pAB3 plasmid and the pBAV shuttle vector did not affect biofilm formation. The A. baumannii ATCC 19606 strain was used as a control. (B) Quantitative determination of biofilm formation. To normalize cell growth rates, A. baumannii strains were grown in LB medium and collected at OD 600 values of 1.0 to 1.2. To count CFU, collected cells were plated on LB medium (top); to quantify the biofilms, absorbance was measured at OD 550 (bottom). Points indicate mean values, and error bars indicate standard deviations of triplicate samples. *, P ≤ 0.002, significant reduction of biofilm production in csuD mutant strains, compared to 17978pAB3−.

    Techniques Used: Crystal Violet Assay, Staining, Mutagenesis, Plasmid Preparation

    31) Product Images from "Outer Membrane Protein, Oma87 Prevents Acinetobacter baumannii Infection"

    Article Title: Outer Membrane Protein, Oma87 Prevents Acinetobacter baumannii Infection

    Journal: International Journal of Peptide Research and Therapeutics

    doi: 10.1007/s10989-020-10056-0

    Survival plot of mice actively immunized with rOma87. The mice were challenged with 2 × LD or 7 × LD of A. baumannii ATCC 19606 and were monitored for 96 h. Survival rates were analyzed using the non-parametric log-rank test (p
    Figure Legend Snippet: Survival plot of mice actively immunized with rOma87. The mice were challenged with 2 × LD or 7 × LD of A. baumannii ATCC 19606 and were monitored for 96 h. Survival rates were analyzed using the non-parametric log-rank test (p

    Techniques Used: Mouse Assay

    Whole cell ELISA with mice sera. Acinetobacter baumannii ATCC 19606 (1 × 10 7 CFU/well) was coated as antigen
    Figure Legend Snippet: Whole cell ELISA with mice sera. Acinetobacter baumannii ATCC 19606 (1 × 10 7 CFU/well) was coated as antigen

    Techniques Used: Enzyme-linked Immunosorbent Assay, Mouse Assay

    Survival plot of mice received 100 µl of immunized or non-immunized mice sera. The mice challenged with 2 × LD or 5 × LD of A. baumannii ATCC 19606. The mice were monitored for 96 h. Survival rates were analyzed using the non-parametric log-rank test (p = 0.002)
    Figure Legend Snippet: Survival plot of mice received 100 µl of immunized or non-immunized mice sera. The mice challenged with 2 × LD or 5 × LD of A. baumannii ATCC 19606. The mice were monitored for 96 h. Survival rates were analyzed using the non-parametric log-rank test (p = 0.002)

    Techniques Used: Mouse Assay

    32) Product Images from "Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections"

    Article Title: Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-03416-2

    ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).
    Figure Legend Snippet: ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).

    Techniques Used: In Silico

    33) Product Images from "Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections"

    Article Title: Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-03416-2

    ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).
    Figure Legend Snippet: ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).

    Techniques Used: In Silico

    34) Product Images from "LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates"

    Article Title: LpxK Is Essential for Growth of Acinetobacter baumannii ATCC 19606: Relationship to Toxic Accumulation of Lipid A Pathway Intermediates

    Journal: mSphere

    doi: 10.1128/mSphere.00199-17

    Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.
    Figure Legend Snippet: Fluorescence microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were grown overnight with IPTG and then subcultured at an appropriate dilution into media without IPTG. Cells were collected after 3 h of growth ( Fig. 1D ) and fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. All figures are to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.
    Figure Legend Snippet: Fluorescence microscopic observation of A. baumannii Δ lpxC cells. A. baumannii ATCC 19606 and A. baumannii Δ lpxC grown to an OD 600 of approximately 0.5 were fixed in 2% glutaraldehyde for microscopy. Green, membrane stained by FM1-43fx; blue, DNA stained by DAPI. The figures are all to the same scale.

    Techniques Used: Fluorescence, Microscopy, Staining

    CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.
    Figure Legend Snippet: CHIR-090, cerulenin, and pyridopyrimidine can rescue the growth of LpxK-depleted cells. (A) JWK0013(pNOV044) was streaked on MHB agar supplemented with 1 mM IPTG and grown overnight at 37°C to induce LpxK expression. The following day, cells were washed repeatedly and resuspended to an OD 600 of 0.01, and a 100-µl volume was plated on MHB agar or MacConkey agar plates without IPTG. Sterile filter discs spotted with IPTG, DMSO, CHIR-090, pyridopyrimidine, or cerulenin were placed on the plates, which were then incubated at 37°C for 24 h (cerulenin was incubated for 72 h). Growth of JWK0013(pNOV044) was restored in the presence of IPTG on both media. Growth of JWK0013(pNOV044) was not observed under noninducing conditions (minus IPTG and DMSO). JWK0013(pNOV044) grew under noninducing conditions in the presence of CHIR-090 (LpxC inhibitor), pyridopyrimidine (acetyl-CoA-carboxylase inhibitor), or cerulenin (β-ketoacyl-acyl carrier protein synthase inhibitor) on MHA but not MacConkey agar. (B) An overnight culture of JWK0013(pNOV044) grown under inducing conditions (+IPTG) was diluted to an OD 600 of 0.1 and then was diluted 100-fold into MHB containing 10% alamarBlue. Next, 100 µl of the inoculum was added to the wells of a 96-well plate containing CHIR-090, pyridopyrimidine, or cerulenin to final assay concentrations ranging from 0 to 128 µg/ml. The plate was incubated for 6 h at 37°C before fluorescence (excitation, 545 nm; emission, 590 nm) was read on a SpectraMax microplate reader, and data were processed with Softmax Pro software v 5.4.1. (C) Cell-associated LOS levels during chemical growth rescue under conditions of LpxK depletion. Lane 1, A. baumannii ATCC 19606 (parent); lane 2, A. baumannii lpxC :: Km r (LOS-deficient cells); lane 3, A. baumannii JWK0013(pNOV044) grown with 1 mM IPTG; lane 4, A. baumannii JWK0013(pNOV044) harvested after grown in the absence of IPTG (LpxK-depleted cells); lane 5, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml CHIR-090; lane 6, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 16 μg/ml pyridopyrimidine; lane 7, A. baumannii JWK0013(pNOV044) grown in the absence of IPTG with 8 μg/ml cerulenin. The LOS gel data are representative of results from three independent experiments.

    Techniques Used: Expressing, Incubation, Fluorescence, Software

    Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.
    Figure Legend Snippet: Transmission electron microscopy of A. baumannii cells depleted for LpxK. A. baumannii ATCC 19606 and JWK0013(pNOV044) were collected as described above for fluorescence microscopy ( Fig. 2 ) and fixed with Tousimis fixative followed by observation using transmission electron microscopy.

    Techniques Used: Transmission Assay, Electron Microscopy, Fluorescence, Microscopy

    35) Product Images from "Relationship between Tolerance and Persistence Mechanisms in Acinetobacter baumannii Strains with AbkAB Toxin-Antitoxin System"

    Article Title: Relationship between Tolerance and Persistence Mechanisms in Acinetobacter baumannii Strains with AbkAB Toxin-Antitoxin System

    Journal: Antimicrobial Agents and Chemotherapy

    doi: 10.1128/AAC.00250-18

    Enzymatic activity by colorimetric assay (WST-1-based) of the isolates A. baumannii ATCC 17978, A. baumannii ATCC 19606, and A. baumannii Ab-2_clon_2010-CHLX in the presence of IMP and CHLX. The x axis represents absorbance (optical density at 450 nm [OD 450 ]), and the y axis represents time (h). *, P
    Figure Legend Snippet: Enzymatic activity by colorimetric assay (WST-1-based) of the isolates A. baumannii ATCC 17978, A. baumannii ATCC 19606, and A. baumannii Ab-2_clon_2010-CHLX in the presence of IMP and CHLX. The x axis represents absorbance (optical density at 450 nm [OD 450 ]), and the y axis represents time (h). *, P

    Techniques Used: Activity Assay, Colorimetric Assay

    Time-kill curves in the presence of antibiotics (IMP) and biocides (CHLX) in susceptible A. baumannii ATCC strains. (a) A. baumannii strain ATCC 17978, which harbors the plasmid with the AbKA/AbkB toxin-antitoxin system (positive control); (b) A. baumannii ATCC 19606 strain without this AbKA/AbkB toxin-antitoxin system (negative control). Box in panel a.3, regrowth is due to putative reactivation of persister cells.
    Figure Legend Snippet: Time-kill curves in the presence of antibiotics (IMP) and biocides (CHLX) in susceptible A. baumannii ATCC strains. (a) A. baumannii strain ATCC 17978, which harbors the plasmid with the AbKA/AbkB toxin-antitoxin system (positive control); (b) A. baumannii ATCC 19606 strain without this AbKA/AbkB toxin-antitoxin system (negative control). Box in panel a.3, regrowth is due to putative reactivation of persister cells.

    Techniques Used: Plasmid Preparation, Positive Control, Negative Control

    36) Product Images from "Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains"

    Article Title: Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains

    Journal: BMC Microbiology

    doi: 10.1186/s12866-020-1722-1

    Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains
    Figure Legend Snippet: Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains

    Techniques Used: Blocking Assay, Expressing

    Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively
    Figure Legend Snippet: Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively

    Techniques Used: Expressing

    Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms
    Figure Legend Snippet: Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms

    Techniques Used: Software

    a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains
    Figure Legend Snippet: a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains

    Techniques Used:

    qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630
    Figure Legend Snippet: qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630

    Techniques Used: Quantitative RT-PCR

    Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline
    Figure Legend Snippet: Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline

    Techniques Used: Transmission Assay

    37) Product Images from "Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains"

    Article Title: Integrative analysis of outer membrane vesicles proteomics and whole-cell transcriptome analysis of eravacycline induced Acinetobacter baumannii strains

    Journal: BMC Microbiology

    doi: 10.1186/s12866-020-1722-1

    Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains
    Figure Legend Snippet: Distribution of genes pertaining to antibiotic resistance and virulence in ATCC 19606 and JU0126 A. baumannii strains. Each block of gradient colors, red (high) to black (low) represents the fold change expression of resistance ( a ) and virulence ( b ) genes from transcriptome analysis of ACC and JU0126 A. baumannii strains

    Techniques Used: Blocking Assay, Expressing

    Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively
    Figure Legend Snippet: Comparison of whole-cell transcriptome and OMV proteome. a A. baumannii ATCC 19606, b A. baumannii JU0126. The log2-fold change represents the ratio of eravacycline treated: control condition. p -value less than 0.05 was considered significant. Genes with no protein expression are considered anticorrelated. Up- and downregulation of genes/proteins are designated depending on positive or negative log2-fold change, respectively

    Techniques Used: Expressing

    Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms
    Figure Legend Snippet: Go enrichment analysis and visualization of genes/proteins from both mRNA and OMVs proteome of A. baumannii ATCC 19606 ( a ) and JU0126 ( b ) strain using ClueGO/CluePedia plug-in from Cytoscape software. The node colors were represented to the biological, molecular and cellular functions of the genes/proteins according to the significant association of related GO terms

    Techniques Used: Software

    a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains
    Figure Legend Snippet: a , b Comparison of differentially expressed genes (DEGs) between eravacycline treated and control samples of A. baumannii ATCC 19606 and clinical strain JU0126. A volcano plot analysis was used to plot the DEGs between control and treated samples of ATCC and JU0126 strains of A. baumannii . Red dots represent upregulated DEGs, green dots represent downregulated DEGs and blue. dots represents no significant change between samples. c , d GO enrichment analysis of differentially expressed genes in eravacycline induced versus control A. baumannii ATCC 19606 and JU0126 strains. The DEGs were categorized into biological (green), cellular (red) and molecular function (blue) components. e , f Scatter plot representation of enriched KEGG pathway statistics of DEGs from A. baumannii ATCC 19606 and JU0126 strains

    Techniques Used:

    qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630
    Figure Legend Snippet: qRT-PCR analyses of six DEGs from each A. baumannii ATCC 19606 and JU0126 strain. Upregulated genes of ATCC 19606- 1: AUO97_00445, 3: AUO97_00560, 5: AUO97_00700. Downregulated genes of ATCC 19606-7: AUO97_05635, 9: AUO97_15195, 11: AUO97_11755. Upregulated genes of JU0126- 2: AUO97_02660, 4: AUO97_03195, 6: AUO97_00745. Downregulated genes of JU0126- 8: AUO97_11755, 10: AUO97_18615, 12: AUO97__18630

    Techniques Used: Quantitative RT-PCR

    Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline
    Figure Legend Snippet: Transmission electron microscopic image of OMVs. a OMVs from A. baumannii ATCC 19606 control strain, b OMVs from A. baumannii ATCC 19606 strain treated with eravacycline, c OMVs from A. baumannii JU0126 control strain, d OMVs from A. baumannii JU0126strain treated with eravacycline

    Techniques Used: Transmission Assay

    38) Product Images from "Effect of Human Burn Wound Exudate on Pseudomonas aeruginosa Virulence"

    Article Title: Effect of Human Burn Wound Exudate on Pseudomonas aeruginosa Virulence

    Journal: mSphere

    doi: 10.1128/mSphere.00111-15

    Growth of burn wound pathogens in burn wound exudate. Growth of E. coli MG1655 X, (×), S. aureus ATCC 29213 (◆) and USA300 (◊), A. baumannii ATCC 19606 ( ─ ), and P. aeruginosa PAO1 (●), PA14 (ο), and PA25688 (▪) was monitored over 24 h in burn wound exudate mix. P. aeruginosa PAO1 growth was also monitored in LB media at pH 7.0 (▴) and pH 9.0 (▵) as a control.
    Figure Legend Snippet: Growth of burn wound pathogens in burn wound exudate. Growth of E. coli MG1655 X, (×), S. aureus ATCC 29213 (◆) and USA300 (◊), A. baumannii ATCC 19606 ( ─ ), and P. aeruginosa PAO1 (●), PA14 (ο), and PA25688 (▪) was monitored over 24 h in burn wound exudate mix. P. aeruginosa PAO1 growth was also monitored in LB media at pH 7.0 (▴) and pH 9.0 (▵) as a control.

    Techniques Used:

    39) Product Images from "Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections"

    Article Title: Constraint-based modeling identifies new putative targets to fight colistin-resistant A. baumannii infections

    Journal: Scientific Reports

    doi: 10.1038/s41598-017-03416-2

    ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).
    Figure Legend Snippet: ( A ) A venn diagram proportionally showing EGs predicted only in Simmons medium (pink), EGs predicted only in rich medium (green), and EGs predicted by both (blue). ( B ) A venn diagram proportionally showing EGs predicted in silico only by iLP844 (pink), EGs obtained only by wet-lab experiment in ATCC 17978 (green), and EGs predicted by both methods (blue). ( C ) A venn diagram proportionally showing essential reactions predicted in iLP844 (pink), essential reactions predicted only in A. baumannii AYE model (AbyMBEL891) (green), and essential reactions predicted by both (blue). ( D ) A venn diagram proportionally showing EGs predicted only in iLP844 (pink), EGs predicted only by wet-lab experiment in A. baumannii ATCC 19606 cell (green), and EGs predicted by both (blue).

    Techniques Used: In Silico

    40) Product Images from "Design of live attenuated bacterial vaccines based on D-glutamate auxotrophy"

    Article Title: Design of live attenuated bacterial vaccines based on D-glutamate auxotrophy

    Journal: Nature Communications

    doi: 10.1038/ncomms15480

    Immunization with D-Glu auxotrophs generates functional and cross-reactive vaccine antisera. ( a ) In vitro OPA and OPKA assays. Per cent killing of A. baumannii ATCC 17978 (4 × 10 4 CFU), ATCC 19606 (6 × 10 4 CFU), AbH12O-A2 (6 × 10 4 CFU) and Ab307-0294 (4 × 10 4 CFU) strains, by α-Ab serum (or naive serum) in the absence or presence of murine RAW 264.7 cells (1.6 × 10 5 ). Per cent killing of P. aeruginosa PAO1 (4 × 10 4 CFU), PA21_ST175 (3 × 10 4 CFU), PA14 (4 × 10 4 CFU) and PA28562 (1 × 10 4 CFU) strains by α-Pa serum (or naive serum) in the absence or presence of human PMNs. Per cent opsonophagocytosis of S. aureus 132 (1.2 × 10 4 CFU), FPR3757 (2.7 × 10 4 CFU), MW2 (1.8 × 10 4 CFU), NEWMAN (1.5 × 10 4 CFU), Sa07997 (3.6 × 10 4 CFU), ED133 (4 × 10 4 CFU) and ED98 (7 × 10 4 CFU) strains by α-Sa serum (or naive serum) in the absence or presence of murine RAW 264.7 cells (1.6 × 10 5 ). * P
    Figure Legend Snippet: Immunization with D-Glu auxotrophs generates functional and cross-reactive vaccine antisera. ( a ) In vitro OPA and OPKA assays. Per cent killing of A. baumannii ATCC 17978 (4 × 10 4 CFU), ATCC 19606 (6 × 10 4 CFU), AbH12O-A2 (6 × 10 4 CFU) and Ab307-0294 (4 × 10 4 CFU) strains, by α-Ab serum (or naive serum) in the absence or presence of murine RAW 264.7 cells (1.6 × 10 5 ). Per cent killing of P. aeruginosa PAO1 (4 × 10 4 CFU), PA21_ST175 (3 × 10 4 CFU), PA14 (4 × 10 4 CFU) and PA28562 (1 × 10 4 CFU) strains by α-Pa serum (or naive serum) in the absence or presence of human PMNs. Per cent opsonophagocytosis of S. aureus 132 (1.2 × 10 4 CFU), FPR3757 (2.7 × 10 4 CFU), MW2 (1.8 × 10 4 CFU), NEWMAN (1.5 × 10 4 CFU), Sa07997 (3.6 × 10 4 CFU), ED133 (4 × 10 4 CFU) and ED98 (7 × 10 4 CFU) strains by α-Sa serum (or naive serum) in the absence or presence of murine RAW 264.7 cells (1.6 × 10 5 ). * P

    Techniques Used: Functional Assay, In Vitro

    Related Articles

    Modification:

    Article Title: New Mutations Involved in Colistin Resistance in Acinetobacter baumannii
    Article Snippet: .. In contrast, resistant strains ATCC 19606 (pmrA P102R , miaA I221V ), ATCC 19606 (pmrA P102R ), ATCC 19606 (pmrA I13M ), and ATCC 19606 (pmrB Q270P ) ( ) displayed not only [M-H]- at m /z 1,910 but also ions at m /z 2,033, which indicates a phosphoethanolamine (pEtN) modification of lipid A. .. DISCUSSION As the last resort for the treatment of multidrug-resistant A. baumannii infection, the mechanism of action of colistin and control of resistance are important for preserving and improving efficacy.

    Concentration Assay:

    Article Title: Colistin-Resistant, Lipopolysaccharide-Deficient Acinetobacter baumannii Responds to Lipopolysaccharide Loss through Increased Expression of Genes Involved in the Synthesis and Transport of Lipoproteins, Phospholipids, and Poly-?-1,6-N-Acetylglucosamine
    Article Snippet: .. LPS-deficient derivatives of strain ATCC 19606 with mutations in any of the lipid A biosynthesis genes, lpxA , lpxC , or lpxD , can occur spontaneously and be selected for in the presence of a high (10 μg/ml) concentration of colistin ( ). .. Furthermore, we have identified a colistin-resistant clinical isolate which has an lpxD mutation and lacks LPS ( ).

    other:

    Article Title: Whole-Genome Sequences of Five Acinetobacter baumannii Strains From a Child With Leukemia M2
    Article Snippet: The five strains were grouped in two clusters, I and II; while, A. baumannii strain ATCC® 19606TM was grouped as an independent cluster.

    Article Title: Polymyxin B in combination with doripenem against heteroresistant Acinetobacter baumannii: pharmacodynamics of new dosing strategies
    Article Snippet: A. baumannii ATCC 19606 (polymyxin B MIC 0.5 mg/L; doripenem MIC 2 mg/L) and a clinical isolate N16870 (polymyxin B MIC 0.5 mg/L; doripenem MIC 16 mg/L) were studied.

    Article Title: Molecular mechanism of antimicrobial activity of chlorhexidine against carbapenem-resistant Acinetobacter baumannii
    Article Snippet: Enhancement of ROS generation in A . baumannii by chlorhexidine Chlorhexidine (32μg/ml) treated A . baumannii strains (RS307 and ATCC-19606) shows the significant elevated ROS generation in the treated bacterial samples as compared to the untreated bacterial samples ( ).

    Positive Control:

    Article Title: Genotyping of carbapenem resistant Acinetobacter baumannii isolated from tracheal tube discharge of hospitalized patients in intensive care units, Ahvaz, Iran
    Article Snippet: .. A . baumannii NCTC 12156 (ATCC 19606) was used as positive control. .. A negative control was included in each PCR reaction, containing all components except the DNA template which was replaced by distilled water ( , ).

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  • 93
    ATCC a baumannii atcc 19606
    Overview of the complete A. <t>baumannii</t> ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).
    A Baumannii Atcc 19606, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 19 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    ATCC a baumannii atcc 19606 genome
    Overview of the complete A. <t>baumannii</t> ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).
    A Baumannii Atcc 19606 Genome, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a baumannii atcc 19606 genome/product/ATCC
    Average 92 stars, based on 2 article reviews
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    92
    ATCC a baumannii atcc 19606t
    Regulation of the uvrABC operon upon exposure of A. <t>baumannii</t> to DNA-damaging agents MMC and UV light. Overnight cultures of A. baumannii <t>ATCC</t> 19606 T carrying any of the three P uvrA fusions (pLPV1Z::P uvrA or pLPV2Z::P uvrA or pLPV3Z::P uvrA ) or the corresponding promoterless vectors were subcultured (1: 100 dilution) in LB broth (pLPV1Z::P uvrA or pLPV2Z::P uvrA ) or M9-S medium (pLPV3Z::P uvrA ) and incubated at 37°C until the culture reached the mid-exponential phase (OD 600 of 0.6). Cultures were treated with different MMC concentrations, as indicated, and luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were measured after 16 h. GFP-producing cells were also visualized using a Leica SP5 confocal laser scanning microscope equipped with a 63× oil immersion objective (D). Representative images of either GFP or GFP and differential interference contrast (DIC)-merged channels are shown. Scale bar, 10 μm. Overnight cultures were also suspended in M9-S medium at an OD 600 of 1.0, and 5 ml of each suspension was irradiated with 0, 10, 50, and 100 J/m 2 UV light at 300 nm. Luminescence (E), β-galactosidase activity (F), and fluorescence emission (G) were measured after a 2-h rescue at 37°C. Gray and black histograms indicate strains carrying individual P uvrA fusions and the promoterless vectors, respectively. Data are the means ± standard deviations from three independent experiments.
    A Baumannii Atcc 19606t, supplied by ATCC, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/a baumannii atcc 19606t/product/ATCC
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    Image Search Results


    Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Article Snippet: A. baumannii ATCC 19606 is one of the best characterised strains in this species ( ).

    Techniques: Plasmid Preparation

    Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Article Snippet: A. baumannii ATCC 19606 is one of the best characterised strains in this species ( ).

    Techniques: Sequencing

    Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Article Snippet: A. baumannii ATCC 19606 is one of the best characterised strains in this species ( ).

    Techniques:

    Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Overview of the complete A. baumannii ATCC 19606 genome. Chromosome: ( A ) From the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG classifications), tRNA (light green), rRNA (dark green), IS elements (dark red), genomic islands (red), prophages (light red), GC content (green) and GC skew (orange for positive and purple for negative). ( B ) Plasmid pMAC: from the outermost to innermost, the tracks indicate the genes on positive strand (dark blue) and negative strand (light blue), ORFs (with colours indicating COG groups), GC content (green) and GC skew (orange for positive and purple for negative).

    Article Snippet: Antibiotic resistance and virulence genes Annotation of A. baumannii ATCC 19606 genome generated 3,805 genes, encoding 3,663 proteins, 74 tRNAs and 4 ncRNA ( ).

    Techniques: Plasmid Preparation

    Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Phylogenetic tree of A. baumannii strains. The ST52 strains ( A. baumannii ATCC 19606 and ab736) are highlighted in violet, whereas IC I and ICII strains are highlighted in red and blue, respectively. The sequence type, GenBank accession, assembly version for each genome are denoted after the strain name.

    Article Snippet: Antibiotic resistance and virulence genes Annotation of A. baumannii ATCC 19606 genome generated 3,805 genes, encoding 3,663 proteins, 74 tRNAs and 4 ncRNA ( ).

    Techniques: Sequencing

    Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Journal: International journal of medical microbiology : IJMM

    Article Title: Complete genome sequence and genome-scale metabolic modelling of Acinetobacter baumannii type strain ATCC 19606

    doi: 10.1016/j.ijmm.2020.151412

    Figure Lengend Snippet: Antimicrobial resistance genes (A) and virulence genes (B) predicted in A. baumannii strains ATCC 19606, AYE, AB5075, MDR-TJ, ACICU, OIFC137, ATCC 17978 and SDF. The red (blue) lattice indicates the presence of antimicrobial resistance and virulence genes, whereas empty lattice indicates absence. The classes of antimicrobial resistance and virulence genes are indicated by colour bars aside. Strains of IC I, II and III are indicated by red, blue and green, respectively.

    Article Snippet: Antibiotic resistance and virulence genes Annotation of A. baumannii ATCC 19606 genome generated 3,805 genes, encoding 3,663 proteins, 74 tRNAs and 4 ncRNA ( ).

    Techniques:

    Regulation of the uvrABC operon upon exposure of A. baumannii to DNA-damaging agents MMC and UV light. Overnight cultures of A. baumannii ATCC 19606 T carrying any of the three P uvrA fusions (pLPV1Z::P uvrA or pLPV2Z::P uvrA or pLPV3Z::P uvrA ) or the corresponding promoterless vectors were subcultured (1: 100 dilution) in LB broth (pLPV1Z::P uvrA or pLPV2Z::P uvrA ) or M9-S medium (pLPV3Z::P uvrA ) and incubated at 37°C until the culture reached the mid-exponential phase (OD 600 of 0.6). Cultures were treated with different MMC concentrations, as indicated, and luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were measured after 16 h. GFP-producing cells were also visualized using a Leica SP5 confocal laser scanning microscope equipped with a 63× oil immersion objective (D). Representative images of either GFP or GFP and differential interference contrast (DIC)-merged channels are shown. Scale bar, 10 μm. Overnight cultures were also suspended in M9-S medium at an OD 600 of 1.0, and 5 ml of each suspension was irradiated with 0, 10, 50, and 100 J/m 2 UV light at 300 nm. Luminescence (E), β-galactosidase activity (F), and fluorescence emission (G) were measured after a 2-h rescue at 37°C. Gray and black histograms indicate strains carrying individual P uvrA fusions and the promoterless vectors, respectively. Data are the means ± standard deviations from three independent experiments.

    Journal: Applied and Environmental Microbiology

    Article Title: New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

    doi: 10.1128/AEM.01334-19

    Figure Lengend Snippet: Regulation of the uvrABC operon upon exposure of A. baumannii to DNA-damaging agents MMC and UV light. Overnight cultures of A. baumannii ATCC 19606 T carrying any of the three P uvrA fusions (pLPV1Z::P uvrA or pLPV2Z::P uvrA or pLPV3Z::P uvrA ) or the corresponding promoterless vectors were subcultured (1: 100 dilution) in LB broth (pLPV1Z::P uvrA or pLPV2Z::P uvrA ) or M9-S medium (pLPV3Z::P uvrA ) and incubated at 37°C until the culture reached the mid-exponential phase (OD 600 of 0.6). Cultures were treated with different MMC concentrations, as indicated, and luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were measured after 16 h. GFP-producing cells were also visualized using a Leica SP5 confocal laser scanning microscope equipped with a 63× oil immersion objective (D). Representative images of either GFP or GFP and differential interference contrast (DIC)-merged channels are shown. Scale bar, 10 μm. Overnight cultures were also suspended in M9-S medium at an OD 600 of 1.0, and 5 ml of each suspension was irradiated with 0, 10, 50, and 100 J/m 2 UV light at 300 nm. Luminescence (E), β-galactosidase activity (F), and fluorescence emission (G) were measured after a 2-h rescue at 37°C. Gray and black histograms indicate strains carrying individual P uvrA fusions and the promoterless vectors, respectively. Data are the means ± standard deviations from three independent experiments.

    Article Snippet: For a preliminary proficiency testing of the performances of pLPV vectors, we investigated the regulation of the DNA damage-inducible uvrABC operon in A. baumannii ATCC 19606T .

    Techniques: Incubation, Activity Assay, Fluorescence, Laser-Scanning Microscopy, Irradiation

    Regulation of the A. baumannii ethanol-inducible adhP (A) and yahK (B) genes. A. baumannii ATCC 19606 T carrying either pLPV1Z::P adhP or pLPV1Z::P yahK was cultured overnight in LB broth, diluted at an OD 600 of 0.1 in LB broth, and challenged with different ethanol concentrations, as indicated. The luminescence emission was recorded at 15 min postinduction. Gray and black histograms indicate pLPV1Z::P adhP (A) or pLPV1Z::P yahK (B) fusions and the promoterless pLPV1Z vector, respectively. Data are the means ± standard deviations from three independent experiments.

    Journal: Applied and Environmental Microbiology

    Article Title: New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

    doi: 10.1128/AEM.01334-19

    Figure Lengend Snippet: Regulation of the A. baumannii ethanol-inducible adhP (A) and yahK (B) genes. A. baumannii ATCC 19606 T carrying either pLPV1Z::P adhP or pLPV1Z::P yahK was cultured overnight in LB broth, diluted at an OD 600 of 0.1 in LB broth, and challenged with different ethanol concentrations, as indicated. The luminescence emission was recorded at 15 min postinduction. Gray and black histograms indicate pLPV1Z::P adhP (A) or pLPV1Z::P yahK (B) fusions and the promoterless pLPV1Z vector, respectively. Data are the means ± standard deviations from three independent experiments.

    Article Snippet: For a preliminary proficiency testing of the performances of pLPV vectors, we investigated the regulation of the DNA damage-inducible uvrABC operon in A. baumannii ATCC 19606T .

    Techniques: Cell Culture, Plasmid Preparation

    Regulation of the basA promoter in response to iron availability. A. baumannii ATCC 19606 T carrying any of the three P basA fusions, namely pLPV1Z::P basA (A), pLPV2Z::P basA (B), or pLPV3Z::P basA (C), or the corresponding promoterless vectors was cultured overnight in LB broth, washed in saline, and suspended at an OD 600 of 0.1 in M9-S medium supplemented with different concentrations of DIP or FeCl 3 , as indicated on the abscissa. Luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were recorded after a 2-h (A and C) or 6-h (B) incubation at 37°C. Gray and black histograms indicate the P basA fusions and the corresponding promoterless vectors, respectively. Dotted lines denote the baseline P basA activity under maximum repression (100 μM FeCl 3 ). Circles indicate bacterial growth (OD 600 ) values without optical path adjustment; the optical path ratio between A or C and B is 0.30. Data are the means ± standard deviations from three independent experiments.

    Journal: Applied and Environmental Microbiology

    Article Title: New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

    doi: 10.1128/AEM.01334-19

    Figure Lengend Snippet: Regulation of the basA promoter in response to iron availability. A. baumannii ATCC 19606 T carrying any of the three P basA fusions, namely pLPV1Z::P basA (A), pLPV2Z::P basA (B), or pLPV3Z::P basA (C), or the corresponding promoterless vectors was cultured overnight in LB broth, washed in saline, and suspended at an OD 600 of 0.1 in M9-S medium supplemented with different concentrations of DIP or FeCl 3 , as indicated on the abscissa. Luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were recorded after a 2-h (A and C) or 6-h (B) incubation at 37°C. Gray and black histograms indicate the P basA fusions and the corresponding promoterless vectors, respectively. Dotted lines denote the baseline P basA activity under maximum repression (100 μM FeCl 3 ). Circles indicate bacterial growth (OD 600 ) values without optical path adjustment; the optical path ratio between A or C and B is 0.30. Data are the means ± standard deviations from three independent experiments.

    Article Snippet: For a preliminary proficiency testing of the performances of pLPV vectors, we investigated the regulation of the DNA damage-inducible uvrABC operon in A. baumannii ATCC 19606T .

    Techniques: Cell Culture, Activity Assay, Fluorescence, Incubation

    In vivo and ex vivo monitoring of A. baumannii iron-regulated gene expression during G. mellonella infection. Larvae were injected with ca. 10 6 cells of A. baumannii ATCC 19606 T carrying either pLPV1Z::P basA or the promoterless pLPV1Z vector and monitored daily for up to 72 h by visual inspection and luminescence imaging of the intact animals (A). Saline injection was used as a control of G. mellonella viability. Luminescence emission by the larvae is shown below the corresponding caterpillars. The hemolymph was sampled from live or agonic caterpillars at 24-h intervals, diluted in saline, and used to determine bacterial viability (B) or luminescence emission (C). Gray and black histograms indicate pLPV1Z::P basA and the corresponding promoterless pLPV1Z vector, respectively. Data are the means ± standard deviations from 5 larvae. (D) Kaplan-Meier curves of the caterpillar population. A log rank test revealed no statistically significant differences between survival rates of larvae injected with A. baumannii ATCC 19606 T carrying either pLPV1Z::P basA or the empty vector, while statistically significant differences were observed between saline- and A. baumannii -injected larvae (****, P

    Journal: Applied and Environmental Microbiology

    Article Title: New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

    doi: 10.1128/AEM.01334-19

    Figure Lengend Snippet: In vivo and ex vivo monitoring of A. baumannii iron-regulated gene expression during G. mellonella infection. Larvae were injected with ca. 10 6 cells of A. baumannii ATCC 19606 T carrying either pLPV1Z::P basA or the promoterless pLPV1Z vector and monitored daily for up to 72 h by visual inspection and luminescence imaging of the intact animals (A). Saline injection was used as a control of G. mellonella viability. Luminescence emission by the larvae is shown below the corresponding caterpillars. The hemolymph was sampled from live or agonic caterpillars at 24-h intervals, diluted in saline, and used to determine bacterial viability (B) or luminescence emission (C). Gray and black histograms indicate pLPV1Z::P basA and the corresponding promoterless pLPV1Z vector, respectively. Data are the means ± standard deviations from 5 larvae. (D) Kaplan-Meier curves of the caterpillar population. A log rank test revealed no statistically significant differences between survival rates of larvae injected with A. baumannii ATCC 19606 T carrying either pLPV1Z::P basA or the empty vector, while statistically significant differences were observed between saline- and A. baumannii -injected larvae (****, P

    Article Snippet: For a preliminary proficiency testing of the performances of pLPV vectors, we investigated the regulation of the DNA damage-inducible uvrABC operon in A. baumannii ATCC 19606T .

    Techniques: In Vivo, Ex Vivo, Expressing, Infection, Injection, Plasmid Preparation, Imaging

    Regulation of the basA promoter in TF-supplemented medium and in HS. A. baumannii ATCC 19606 T carrying any of the three P basA fusions (pLPV1Z::P basA or pLPV2Z::P basA or pLPV3Z::P basA ) or the corresponding promoterless vector was grown overnight in LB broth, washed with saline, and inoculated (OD 600 of 0.1) in either M9-S medium supplemented with 2.5 mg/ml TF (TF) or in HS (HS). As a control, 100 μM FeCl 3 was added to TF (TF + 100 μM Fe) or HS (HS + 100 μM Fe) to saturate the TF iron binding capacity. Luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were recorded after a 4-h (A to C) or 6-h (D to F) incubation at 37°C. Gray and black histograms indicate the P basA fusions and the corresponding promoterless vectors, respectively. Data are the means ± standard deviations from three independent experiments.

    Journal: Applied and Environmental Microbiology

    Article Title: New Shuttle Vectors for Real-Time Gene Expression Analysis in Multidrug-Resistant Acinetobacter Species: In Vitro and In Vivo Responses to Environmental Stressors

    doi: 10.1128/AEM.01334-19

    Figure Lengend Snippet: Regulation of the basA promoter in TF-supplemented medium and in HS. A. baumannii ATCC 19606 T carrying any of the three P basA fusions (pLPV1Z::P basA or pLPV2Z::P basA or pLPV3Z::P basA ) or the corresponding promoterless vector was grown overnight in LB broth, washed with saline, and inoculated (OD 600 of 0.1) in either M9-S medium supplemented with 2.5 mg/ml TF (TF) or in HS (HS). As a control, 100 μM FeCl 3 was added to TF (TF + 100 μM Fe) or HS (HS + 100 μM Fe) to saturate the TF iron binding capacity. Luminescence (A), β-galactosidase activity (B), and fluorescence emission (C) were recorded after a 4-h (A to C) or 6-h (D to F) incubation at 37°C. Gray and black histograms indicate the P basA fusions and the corresponding promoterless vectors, respectively. Data are the means ± standard deviations from three independent experiments.

    Article Snippet: For a preliminary proficiency testing of the performances of pLPV vectors, we investigated the regulation of the DNA damage-inducible uvrABC operon in A. baumannii ATCC 19606T .

    Techniques: Plasmid Preparation, Binding Assay, Activity Assay, Fluorescence, Incubation