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  • 99
    Thermo Fisher brain heart infusion bhi
    Brain Heart Infusion Bhi, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 318 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore ly bhi
    Ly Bhi, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco brain heart infusion bhi difco
    Growth patterns of E. <t>faecalis</t> strains in <t>BHI</t> and BHIS. Data from BHI grown cultures were marked with solid squares, circles and triangles and BHIS were marked with open squares, circles and triangles. Time points at which adherence was tested are marked
    Brain Heart Infusion Bhi Difco, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 87 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco brain heart infusion broth bhi difco
    Protease activation in the extracellular protein fraction from P. <t>gingivalis</t> FLL93. P. gingivalis was grown to stationary phase (OD 600 of 1.3) in 1 liter of <t>BHI</t> broth supplemented with hemin and vitamin K. Acetone (37.5%)-precipitated proteins were mixed with 8 M urea and were incubated at 4°C for 1 h. The urea was slowly removed from the mixture by centrifugation (10,000 × g ) in a Millipore filtration unit with the addition of increasing volumes of 100 mM Tris-HCl buffer (pH 7.4). Activities for Rgp and Kgp were tested. Error bars indicate the standard errors of the means of three independent trials.
    Brain Heart Infusion Broth Bhi Difco, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 35 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco brain heart infusion medium bhi difco
    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. <t>monocytogenes</t> to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in <t>BHI</t> were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p
    Brain Heart Infusion Medium Bhi Difco, supplied by Difco, used in various techniques. Bioz Stars score: 85/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson brain heart infusion bhi bd difco tv kan plates
    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. <t>monocytogenes</t> to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in <t>BHI</t> were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p
    Brain Heart Infusion Bhi Bd Difco Tv Kan Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 85/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson brain heart infusion bhi
    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. <t>monocytogenes</t> to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in <t>BHI</t> were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p
    Brain Heart Infusion Bhi, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 592 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson brain heart infusion bhi broth
    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. <t>monocytogenes</t> to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in <t>BHI</t> were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p
    Brain Heart Infusion Bhi Broth, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 1043 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco brain heart infusion bhi medim
    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. <t>monocytogenes</t> to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in <t>BHI</t> were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p
    Brain Heart Infusion Bhi Medim, supplied by Difco, used in various techniques. Bioz Stars score: 88/100, based on 46 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson brain heart infusion bhi medium
    Measurement of extracellular pyruvate during growth of S. <t>mutans</t> UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ) and Δ pta Δ ackA ( d ) strains were grown in <t>BHI.</t> For time course measurements of extracellular pyruvate and growth, samples were taken at 1 or 2 h intervals (see Materials and Methods for details). The concentration of pyruvate was determined using an EnzyChrom™ pyruvate assay kit, and growth was measured by the optical density at 600 nm (OD 600 ). Bars indicate the average concentration of extracellular pyruvate and a solid line with black circles indicates the corresponding growth curve. The results are an average of two independent experiments. Error bars = standard deviation.
    Brain Heart Infusion Bhi Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 362 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco bactotm brain heart infusion bhi broth
    Measurement of extracellular pyruvate during growth of S. <t>mutans</t> UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ) and Δ pta Δ ackA ( d ) strains were grown in <t>BHI.</t> For time course measurements of extracellular pyruvate and growth, samples were taken at 1 or 2 h intervals (see Materials and Methods for details). The concentration of pyruvate was determined using an EnzyChrom™ pyruvate assay kit, and growth was measured by the optical density at 600 nm (OD 600 ). Bars indicate the average concentration of extracellular pyruvate and a solid line with black circles indicates the corresponding growth curve. The results are an average of two independent experiments. Error bars = standard deviation.
    Bactotm Brain Heart Infusion Bhi Broth, supplied by Difco, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Growth patterns of E. faecalis strains in BHI and BHIS. Data from BHI grown cultures were marked with solid squares, circles and triangles and BHIS were marked with open squares, circles and triangles. Time points at which adherence was tested are marked

    Journal:

    Article Title: Role of Serum, a Biological Cue, in Adherence of Enterococcus faecalis to Extracellular Matrix Proteins, Collagen, Fibrinogen, and Fibronectin

    doi: 10.1086/588143

    Figure Lengend Snippet: Growth patterns of E. faecalis strains in BHI and BHIS. Data from BHI grown cultures were marked with solid squares, circles and triangles and BHIS were marked with open squares, circles and triangles. Time points at which adherence was tested are marked

    Article Snippet: E. faecalis cells were grown in Brain Heart Infusion (BHI) (Difco) at 37°C for routine purposes.

    Techniques:

    pPplA enhances bacterial aggregation in broth culture. (A) Image of bacterial aggregation observed between the wild-type L . monocytogenes 10403S strain versus a prfA * mutant when bacterial cultures grown in BHI are left statically overnight at room-temperature (RT). (B) Measurement of the rate of bacterial aggregation in BHI. The optical-density at 600nm was monitored at the indicated time points for 1 mL of an overnight culture initially grown in BHI with shaking at 37°C then left statically at RT, where bacterial aggregation is measured as the decrease in the optical-density of the culture supernatant as the bacterial aggregate out of solution. (C) Measurement of bacterial aggregation of the indicated mutant strains resuspended in 1 mL of spent media derived from overnight stationary phase cultures either containing the pPplA peptide (G72stop) or lacking it (Δ pplA ). The ability of the pPplA containing media (G72stop) to restore bacterial aggregation indicates the presence of a secreted substance (potentially pPplA) that enhances bacterial aggregation in broth culture. (D) Measurement of bacterial aggregation as done in panel C, except strains were resuspended in 1 mL of BHI spent media derived from E . coli containing the complementation vector construct expressing the N-terminal 72 amino acids of PplA as described in Fig. 2A or the empty vector. The ability of spent media derived from an E . coli strain containing the first 72 amino acids of pPplA supports the secretion of a PplA N-terminus derived peptide that enhances bacterial aggregation. (E) Assessment of bacterial aggregation in a strain containing three amino acid substitutions within the predicted peptide sequence (referred to as prfA * pplA m ). Reduced aggregation of prfA * pplA m indicates the importance of these amino acids within the pPplA pheromone. (F) Bacterial aggregation of two oligopeptide transport mutants, a prfA * Δ ctaP compared to a prfA *- oppA insertion mutant. A prfA * Δ ctaP is impaired for bacterial aggregation, indicating a possible link between the pPplA peptide and import of the peptide through the CtaP transport system. For panels (B), (C) and (D), data is representative of at least three independent experiments.

    Journal: PLoS Pathogens

    Article Title: Identification of a Peptide-Pheromone that Enhances Listeria monocytogenes Escape from Host Cell Vacuoles

    doi: 10.1371/journal.ppat.1004707

    Figure Lengend Snippet: pPplA enhances bacterial aggregation in broth culture. (A) Image of bacterial aggregation observed between the wild-type L . monocytogenes 10403S strain versus a prfA * mutant when bacterial cultures grown in BHI are left statically overnight at room-temperature (RT). (B) Measurement of the rate of bacterial aggregation in BHI. The optical-density at 600nm was monitored at the indicated time points for 1 mL of an overnight culture initially grown in BHI with shaking at 37°C then left statically at RT, where bacterial aggregation is measured as the decrease in the optical-density of the culture supernatant as the bacterial aggregate out of solution. (C) Measurement of bacterial aggregation of the indicated mutant strains resuspended in 1 mL of spent media derived from overnight stationary phase cultures either containing the pPplA peptide (G72stop) or lacking it (Δ pplA ). The ability of the pPplA containing media (G72stop) to restore bacterial aggregation indicates the presence of a secreted substance (potentially pPplA) that enhances bacterial aggregation in broth culture. (D) Measurement of bacterial aggregation as done in panel C, except strains were resuspended in 1 mL of BHI spent media derived from E . coli containing the complementation vector construct expressing the N-terminal 72 amino acids of PplA as described in Fig. 2A or the empty vector. The ability of spent media derived from an E . coli strain containing the first 72 amino acids of pPplA supports the secretion of a PplA N-terminus derived peptide that enhances bacterial aggregation. (E) Assessment of bacterial aggregation in a strain containing three amino acid substitutions within the predicted peptide sequence (referred to as prfA * pplA m ). Reduced aggregation of prfA * pplA m indicates the importance of these amino acids within the pPplA pheromone. (F) Bacterial aggregation of two oligopeptide transport mutants, a prfA * Δ ctaP compared to a prfA *- oppA insertion mutant. A prfA * Δ ctaP is impaired for bacterial aggregation, indicating a possible link between the pPplA peptide and import of the peptide through the CtaP transport system. For panels (B), (C) and (D), data is representative of at least three independent experiments.

    Article Snippet: L . monocytogenes and E . coli strains were grown at 37°C in brain heart infusion (BHI) media (Difco Laboratories, Detroit, MI) and Luria broth (LB) (Invitrogen Corp., Carlsbad, CA).

    Techniques: Mutagenesis, Derivative Assay, Plasmid Preparation, Construct, Expressing, Sequencing

    Construction of L . monocytogenes mutants that lack the PplA lipoprotein but retain peptide pheromone secretion. (A) Strategy for the construction of the in-frame pplA deletion and pplA -G72stop mutants that retain peptide pheromone secretion. The region used for complementation of the pplA deletion is indicated by the dashed lines. The C terminal lipoprotein region of PplA expressed and purified from E . coli that was used for affinity purification of the PplA antibody and also as the (+) control in western blots is indicated by the blue solid line. (B) Assessment of growth of the lipoprotein and pheromone mutants in BHI broth culture media. Overnight cultures of each strain grown shaking in BHI at 37°C were diluted 1:20 in fresh BHI media and the OD 600 was measured at the indicated time points. (C) Western blot analysis of both surface-associated and secreted PlpA lipoprotein isolated from stationary phase cultures grown overnight shaking at 37°C. Samples were normalized to OD 600 . Secreted PplA was TCA extracted from the culture supernatant and surface-associated PplA was isolated by boiling in SDS-boiling buffer. Arrow indicates the position of full length PplA. ‘*’ indicates truncated and purified PplA lipoprotein used as positive control for antibody recognition. PplA lipoprotein is primarily detected in the surface-associated preparations versus the supernatant of wild-type L . monocytogenes . For panels (B) and (C), data is representative of at least three-independent experiments.

    Journal: PLoS Pathogens

    Article Title: Identification of a Peptide-Pheromone that Enhances Listeria monocytogenes Escape from Host Cell Vacuoles

    doi: 10.1371/journal.ppat.1004707

    Figure Lengend Snippet: Construction of L . monocytogenes mutants that lack the PplA lipoprotein but retain peptide pheromone secretion. (A) Strategy for the construction of the in-frame pplA deletion and pplA -G72stop mutants that retain peptide pheromone secretion. The region used for complementation of the pplA deletion is indicated by the dashed lines. The C terminal lipoprotein region of PplA expressed and purified from E . coli that was used for affinity purification of the PplA antibody and also as the (+) control in western blots is indicated by the blue solid line. (B) Assessment of growth of the lipoprotein and pheromone mutants in BHI broth culture media. Overnight cultures of each strain grown shaking in BHI at 37°C were diluted 1:20 in fresh BHI media and the OD 600 was measured at the indicated time points. (C) Western blot analysis of both surface-associated and secreted PlpA lipoprotein isolated from stationary phase cultures grown overnight shaking at 37°C. Samples were normalized to OD 600 . Secreted PplA was TCA extracted from the culture supernatant and surface-associated PplA was isolated by boiling in SDS-boiling buffer. Arrow indicates the position of full length PplA. ‘*’ indicates truncated and purified PplA lipoprotein used as positive control for antibody recognition. PplA lipoprotein is primarily detected in the surface-associated preparations versus the supernatant of wild-type L . monocytogenes . For panels (B) and (C), data is representative of at least three-independent experiments.

    Article Snippet: L . monocytogenes and E . coli strains were grown at 37°C in brain heart infusion (BHI) media (Difco Laboratories, Detroit, MI) and Luria broth (LB) (Invitrogen Corp., Carlsbad, CA).

    Techniques: Purification, Affinity Purification, Western Blot, Isolation, Positive Control

    Growth of mutant H. influenzae Rd strains. The strains Rd, REI1012 (Δ hel :: kan ), and GK04 ( nadN :: cat Δ hel :: kan ) were grown on BHI agar plates supplemented with hemin (20 μg/ml) and different nicotinamide nucleotide concentrations and sources. Growth phenotype is shown with 1.5 and 35 μM NAD, with 1.5 and 15 μM NMN, and with 1.5 and 15 μM NR, as indicated.

    Journal: Journal of Bacteriology

    Article Title: NadN and e (P4) Are Essential for Utilization of NAD and Nicotinamide Mononucleotide but Not Nicotinamide Riboside in Haemophilus influenzae

    doi: 10.1128/JB.183.13.3974-3981.2001

    Figure Lengend Snippet: Growth of mutant H. influenzae Rd strains. The strains Rd, REI1012 (Δ hel :: kan ), and GK04 ( nadN :: cat Δ hel :: kan ) were grown on BHI agar plates supplemented with hemin (20 μg/ml) and different nicotinamide nucleotide concentrations and sources. Growth phenotype is shown with 1.5 and 35 μM NAD, with 1.5 and 15 μM NMN, and with 1.5 and 15 μM NR, as indicated.

    Article Snippet: H. influenzae was grown at 37°C under aerobic conditions on 3.8% brain heart infusion (BHI) agar (Difco Laboratories, Detroit, Mich.) supplemented with NAD (15 to 30 μM) and hemin-chloride (20 μg/ml) (Sigma, Deisenhofen, Germany).

    Techniques: Mutagenesis

    Protease activation in the extracellular protein fraction from P. gingivalis FLL93. P. gingivalis was grown to stationary phase (OD 600 of 1.3) in 1 liter of BHI broth supplemented with hemin and vitamin K. Acetone (37.5%)-precipitated proteins were mixed with 8 M urea and were incubated at 4°C for 1 h. The urea was slowly removed from the mixture by centrifugation (10,000 × g ) in a Millipore filtration unit with the addition of increasing volumes of 100 mM Tris-HCl buffer (pH 7.4). Activities for Rgp and Kgp were tested. Error bars indicate the standard errors of the means of three independent trials.

    Journal: Infection and Immunity

    Article Title: The vimE Gene Downstream of vimA Is Independently Expressed and Is Involved in Modulating Proteolytic Activity in Porphyromonas gingivalis W83

    doi: 10.1128/IAI.72.10.5555-5564.2004

    Figure Lengend Snippet: Protease activation in the extracellular protein fraction from P. gingivalis FLL93. P. gingivalis was grown to stationary phase (OD 600 of 1.3) in 1 liter of BHI broth supplemented with hemin and vitamin K. Acetone (37.5%)-precipitated proteins were mixed with 8 M urea and were incubated at 4°C for 1 h. The urea was slowly removed from the mixture by centrifugation (10,000 × g ) in a Millipore filtration unit with the addition of increasing volumes of 100 mM Tris-HCl buffer (pH 7.4). Activities for Rgp and Kgp were tested. Error bars indicate the standard errors of the means of three independent trials.

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (both from Difco Laboratories, Detroit, Mich.), hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (0.1%) (all from Sigma-Aldrich, St. Louis, Mo.).

    Techniques: Activation Assay, Incubation, Centrifugation, Filtration

    Inactivation of vimE resulted in decreased proteolytic activity of P. gingivalis. P. gingivalis was grown to late log phase (OD 600 of 0.7 to 0.8) or stationary phase (OD 600 of 1.4 to 1.5) in BHI broth supplemented with yeast extract, hemin, and vitamin K. Activities against Rgp (A) and Kgp (B) were tested in whole-cell culture. The results shown are representative of three independent experiments in triplicate. E, exponential phase; L, stationary phase. Error bars indicate the standard errors of the means of three independent trials.

    Journal: Infection and Immunity

    Article Title: The vimE Gene Downstream of vimA Is Independently Expressed and Is Involved in Modulating Proteolytic Activity in Porphyromonas gingivalis W83

    doi: 10.1128/IAI.72.10.5555-5564.2004

    Figure Lengend Snippet: Inactivation of vimE resulted in decreased proteolytic activity of P. gingivalis. P. gingivalis was grown to late log phase (OD 600 of 0.7 to 0.8) or stationary phase (OD 600 of 1.4 to 1.5) in BHI broth supplemented with yeast extract, hemin, and vitamin K. Activities against Rgp (A) and Kgp (B) were tested in whole-cell culture. The results shown are representative of three independent experiments in triplicate. E, exponential phase; L, stationary phase. Error bars indicate the standard errors of the means of three independent trials.

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (both from Difco Laboratories, Detroit, Mich.), hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (0.1%) (all from Sigma-Aldrich, St. Louis, Mo.).

    Techniques: Activity Assay, Cell Culture

    Immunoreactivity of the RgpB proenzyme in P. gingivalis FLL93. Western blot analysis using specific RgpB proenzyme antibodies as probes was done on the extracellular fractions from P. gingivalis W83 (lane 1), FLL92 (lane 2), and FLL93 (lane 3). All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at exponential growth phase.

    Journal: Infection and Immunity

    Article Title: The vimE Gene Downstream of vimA Is Independently Expressed and Is Involved in Modulating Proteolytic Activity in Porphyromonas gingivalis W83

    doi: 10.1128/IAI.72.10.5555-5564.2004

    Figure Lengend Snippet: Immunoreactivity of the RgpB proenzyme in P. gingivalis FLL93. Western blot analysis using specific RgpB proenzyme antibodies as probes was done on the extracellular fractions from P. gingivalis W83 (lane 1), FLL92 (lane 2), and FLL93 (lane 3). All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at exponential growth phase.

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (both from Difco Laboratories, Detroit, Mich.), hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (0.1%) (all from Sigma-Aldrich, St. Louis, Mo.).

    Techniques: Western Blot

    Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. monocytogenes to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in BHI were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p

    Journal: PLoS Pathogens

    Article Title: Cyclic di-GMP-dependent Signaling Pathways in the Pathogenic Firmicute Listeria monocytogenes

    doi: 10.1371/journal.ppat.1004301

    Figure Lengend Snippet: Role of the c-di-GMP-induced EPS in biofilm formation, cell aggregation, and tolerance of L. monocytogenes to disinfectants and desiccation. A : Biofilm formation of L. monocytogenes in 96-well polystyrene plates (measured using a Crystal violet dye-binding assay). Cultures were grown for 6 days at 30°C in LB (top panel) or LB supplemented with 3% glycerol (bottom panel). Shown are average results from two biological replicates, where each strain was grown in six wells in a replicate (i.e., six technical replicates). Black circle, wild type; red square, Δ pdeB/C/D ; green triangle, Δ pdeB/C/D Δ pssC ; blue cross, Δ pdeB/C/D Δ pssE . B : EPS-dependent L. monocytogenes cell aggregation (clumping) in HTM medium. Overnight cultures grown in BHI were inoculated into HTM liquid medium at A 600 of 0.01 and incubated at 30°C with gentle shaking (rotary shaker, 125 rpm) for 48 h. 1, ΔpdeB/C/D ; 2, wild type; 3, ΔpdeB/C/D ΔpssC ; 4, ΔpdeB/C/D ΔpssE ; C : Protective role of the c-di-GMP-inducible EPS in disinfection. Aliquots of the HTM-grown cultures were mixed with disinfectant solutions for 10 min at room temperature. Disinfection was stopped by adding a D/E neutralizing broth (Difco); the cultures were vortexed vigorously (5 min) with glass beads to break clumps and plated on BHI agar. Colonies were enumerated after a 48-h growth at 37°C. SH, sodium hydrochloride (1600 ppm); HP, hydrogen peroxide (200 mM); BC, benzalkonium chloride (100 ppm). White background, EGD-e; black, ΔpdeB/C/D ; grey, ΔpdeB/C/D ΔpssC . SH, sodium hypochlorite; HP, hydrogen peroxide; BC, benzalkonium chloride. The absence of the bar for the EGD-e strain treated with SH indicates the lack of survivors. D : Protective role of the c-di-GMP-inducible EPS in desiccation. Aliquots of overnight cultures grown in HTM at 37°C were spun down, the supernatants were removed, and cell pellets were stored in desiccators at room temperature for the indicated periods. The pellets were rehydrated, vortexed with glass beads for better suspension and plated on BHI agar. The numbers of surviving colonies after incubation at 37°C for 24 h are plotted. In panels C and D, bars denote mean values for data from three biological replicates. *, significantly different (p

    Article Snippet: L. monocytogenes was grown in Brain Heart Infusion (BHI) medium (Difco), HTM (minimal medium containing 3% glucose) or LB, supplemented with appropriate antibiotics at 25, 30, 37 or 42°C, as indicated.

    Techniques: Binding Assay, Incubation

    Measurement of extracellular pyruvate during growth of S. mutans UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ) and Δ pta Δ ackA ( d ) strains were grown in BHI. For time course measurements of extracellular pyruvate and growth, samples were taken at 1 or 2 h intervals (see Materials and Methods for details). The concentration of pyruvate was determined using an EnzyChrom™ pyruvate assay kit, and growth was measured by the optical density at 600 nm (OD 600 ). Bars indicate the average concentration of extracellular pyruvate and a solid line with black circles indicates the corresponding growth curve. The results are an average of two independent experiments. Error bars = standard deviation.

    Journal: Microorganisms

    Article Title: The Pta-AckA Pathway Regulates LrgAB-Mediated Pyruvate Uptake in Streptococcus mutans

    doi: 10.3390/microorganisms8060846

    Figure Lengend Snippet: Measurement of extracellular pyruvate during growth of S. mutans UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ) and Δ pta Δ ackA ( d ) strains were grown in BHI. For time course measurements of extracellular pyruvate and growth, samples were taken at 1 or 2 h intervals (see Materials and Methods for details). The concentration of pyruvate was determined using an EnzyChrom™ pyruvate assay kit, and growth was measured by the optical density at 600 nm (OD 600 ). Bars indicate the average concentration of extracellular pyruvate and a solid line with black circles indicates the corresponding growth curve. The results are an average of two independent experiments. Error bars = standard deviation.

    Article Snippet: S. mutans UA159 and its derivative strains were grown in brain heart infusion (BHI) medium (BD Difco™, Franklin, NJ, USA) as overnight static cultures at 37 °C in a 5% CO2 atmosphere.

    Techniques: Concentration Assay, Pyruvate Assay, Standard Deviation

    The contribution of the Pfl pathway to stationary phase P lrgA activity ( a – d ) and pyruvate uptake for regrowth ( e – h ). For measurement of P lrgA activation, the P lrgA-gfp reporter strain in the wild type UA159 ( a ), Δ pfl ( b ), Δ pfl2 ( c ) and Δ pflA ( d ) backgrounds, was grown in FMC11 medium. Relative gfp expression (green circle) and cell growth (OD 600 ; white diamond) were monitored during growth on a plate reader (see Materials and Methods for details). For measurement of growth in response to external pyruvate, wild type UA159 ( e ), Δ pfl ( f ), Δ pfl2 ( g ) and Δ pflA strains were grown in BHI, supplemented by different concentrations of pyruvate (0, 10 and 40 mM). Optical density at 600 nm was monitored every 30 min at 37 °C using the Bioscreen C lab system. The results are representative of three independent experiments.

    Journal: Microorganisms

    Article Title: The Pta-AckA Pathway Regulates LrgAB-Mediated Pyruvate Uptake in Streptococcus mutans

    doi: 10.3390/microorganisms8060846

    Figure Lengend Snippet: The contribution of the Pfl pathway to stationary phase P lrgA activity ( a – d ) and pyruvate uptake for regrowth ( e – h ). For measurement of P lrgA activation, the P lrgA-gfp reporter strain in the wild type UA159 ( a ), Δ pfl ( b ), Δ pfl2 ( c ) and Δ pflA ( d ) backgrounds, was grown in FMC11 medium. Relative gfp expression (green circle) and cell growth (OD 600 ; white diamond) were monitored during growth on a plate reader (see Materials and Methods for details). For measurement of growth in response to external pyruvate, wild type UA159 ( e ), Δ pfl ( f ), Δ pfl2 ( g ) and Δ pflA strains were grown in BHI, supplemented by different concentrations of pyruvate (0, 10 and 40 mM). Optical density at 600 nm was monitored every 30 min at 37 °C using the Bioscreen C lab system. The results are representative of three independent experiments.

    Article Snippet: S. mutans UA159 and its derivative strains were grown in brain heart infusion (BHI) medium (BD Difco™, Franklin, NJ, USA) as overnight static cultures at 37 °C in a 5% CO2 atmosphere.

    Techniques: Activity Assay, Activation Assay, Expressing

    The effect of exogenously added pyruvate on the stationary phase of growth of S. mutans UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ), KB12 ( pta -complemented, ( d ) and KB034 ( ackA -complemented, ( e ) strains were grown in BHI, supplemented by different concentrations of pyruvate (0, 10 and 40 mM). Optical density at 600 nm was monitored every 30 min at 37 °C using the Bioscreen C lab system. The results are representative of three independent experiments.

    Journal: Microorganisms

    Article Title: The Pta-AckA Pathway Regulates LrgAB-Mediated Pyruvate Uptake in Streptococcus mutans

    doi: 10.3390/microorganisms8060846

    Figure Lengend Snippet: The effect of exogenously added pyruvate on the stationary phase of growth of S. mutans UA159 wild type and Pta-AckA pathway derivative strains. Wild type ( a ), Δ pta ( b ), Δ ackA ( c ), KB12 ( pta -complemented, ( d ) and KB034 ( ackA -complemented, ( e ) strains were grown in BHI, supplemented by different concentrations of pyruvate (0, 10 and 40 mM). Optical density at 600 nm was monitored every 30 min at 37 °C using the Bioscreen C lab system. The results are representative of three independent experiments.

    Article Snippet: S. mutans UA159 and its derivative strains were grown in brain heart infusion (BHI) medium (BD Difco™, Franklin, NJ, USA) as overnight static cultures at 37 °C in a 5% CO2 atmosphere.

    Techniques: