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  • 99
    Millipore middlebrook 7h10 agar
    Middlebrook 7h10 Agar, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher middlebrook 7h10
    garA is required for growth of M . tuberculosis in vitro , survival in macrophages, and virulence in mice. ( A ) M . tuberculosis lacking garA was unable to grow on standard <t>7H10</t> medium unless supplemented with asparagine. Plasmid-borne garA restored the defect, but variants of garA with mutations at threonine 21 in the phosphorylation motif (ETTS) gave only partial complementation. Strains were grown in <t>Middlebrook</t> 7H9 plus 30 mM asparagine then washed and diluted in standard 7H9 and spotted onto standard 7H10 with or without 10 mM asparagine. Photographs are representative of at least 3 independent experiments. ( B ) M . tuberculosis lacking garA (red squares) had a defect in growth and survival in differentiated THP-1 cells compared to parental M . tuberculosis H37Rv (black circles). Re-introduction of GarA (black triangles) or variants of GarA lacking a single phosphorylation site (grey crosses and squares) restored growth but variant GarA lacking both phosphorylation sites (green triangles) did not. Data points show the mean and standard deviation from four replicates and are representative of two independent experiments. ( C ) M . tuberculosis lacking garA was avirulent in mice as it was eliminated from the lungs. BALB/C mice were infected intranasally with 10 5 bacilli and bacterial burden was measured on days 1, 7, 21 and 28. Data points show the bacterial burden in individual animals. The bacterial burden of mice infected with Δ garA Mt (red squares) was significantly lower than those infected with M . tuberculosis H37Rv (black circles), or complemented Δ garA Mt (black triangles) at all time points from day 7 (p
    Middlebrook 7h10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals middlebrook mb 7h10
    Growth of M. smegmatis mc 2 155 and M371 under fluoroquinolones exposure. Ten-fold serial dilutions of wild-type, M371, M371-pALACE- pafC and M371-pALACE were spotted on <t>Middlebrook</t> <t>7H10</t> containing indicated concentration of moxifloxacin, norfloxacin, ofloxacin and ciprofloxacin. Then the result was recorded when incubated at 37 °C for 3 days.
    Middlebrook Mb 7h10, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 90/100, based on 21 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco middlebrook 7h10
    mc 2 7901 and mc 2 7902 grow similarly to virulent M. tuberculosis in vitro . (A) Log-phase cultures of mc 2 7901 and mc 2 7902 grown in <t>Middlebrook</t> 7H9-OADC-glycerol-tyloxapol-PLAM were diluted 1/100, and growth was followed by recording optical density at 600 nm (OD 600 ) over time. Mean with standard deviation is plotted ( n = 2). (B) RAW 264.7 macrophages were infected at an MOI of 1 with mc 2 7901, mc 2 7902, or H37Rv. At the indicated time points, macrophages were lysed, and bacterial titers were determined by plating for CFU on Middlebrook <t>7H10-OADC-glycerol-PLAM</t> plates. PLAM was added to the macrophage growth medium, and the medium was changed at each time point. (C) Growth of mc 2 7901 and mc 2 7902 in RAW 264.7 macrophages relative to the inocula (same experiment as in panel B). Mean with standard deviation is plotted ( n = 2).
    Middlebrook 7h10, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 179 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10
    Survival of M. tuberculosis ( Mtb ) inside human macrophages. Human MDMs infected with Mtb H37Rv (MOI 5). The percentages of intracellular and not phagocytosed bacilli immediately after infection were determined by CFU assay and compared to inoculum (a). Lysed infected macrophages and released bacteria in the culture supernatants at 1, 3, 7 and 11 days after infection were plated on <t>Middlebrook</t> <t>7H10</t> with OADC and the CFU were counted after 14 days (b). Cytotoxicity induced by Mtb in human MDMs was measured by LDH activity (c). Values are the means ± SD from four independent experiments. The percentage of infected cells was measured, after 3 hr of infection (time 0) and 1, 3, 7 days after infection, by acridine-orange staining (d). and the representative pictures of infected MDMs, are shown (e). A significant difference was detected with respect to 1 and 3 days post-infection (* P
    Middlebrook 7h10, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 141 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HiMedia Laboratories middlebrook 7h10
    Survival of M. tuberculosis ( Mtb ) inside human macrophages. Human MDMs infected with Mtb H37Rv (MOI 5). The percentages of intracellular and not phagocytosed bacilli immediately after infection were determined by CFU assay and compared to inoculum (a). Lysed infected macrophages and released bacteria in the culture supernatants at 1, 3, 7 and 11 days after infection were plated on <t>Middlebrook</t> <t>7H10</t> with OADC and the CFU were counted after 14 days (b). Cytotoxicity induced by Mtb in human MDMs was measured by LDH activity (c). Values are the means ± SD from four independent experiments. The percentage of infected cells was measured, after 3 hr of infection (time 0) and 1, 3, 7 days after infection, by acridine-orange staining (d). and the representative pictures of infected MDMs, are shown (e). A significant difference was detected with respect to 1 and 3 days post-infection (* P
    Middlebrook 7h10, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco middlebrook 7h10 agar
    Effects of euglycaemic and hyperglycaemic culture conditions on H37RV infection and cytokine production in vitro. (A) CD14 + selected monocytes were differentiated into M2 macrophages in the presence of 5 mmol/L glucose, 5 mmol/L glucose and 20 mmol/L mannitol or 25 mmol/L glucose. The macrophages were infected for 1 hour with H37Rv at an MOI of 10. After infection the macrophages were washed and fresh media containing the different glucose media was added. After 24 hours supernatants were collected and the cells were lysed by osmotic pressure. (B) Cell lysates were serially diluted and plated on <t>Middlebrook</t> <t>7H10</t> agar. CFUs were counted after 2–3 weeks of growth at 37°C. Data are shown as mean ± SEM of two independent experiments (n = 4). (C) Pro-inflammatory cytokines TNF-α and IL-6 were measured along with the anti-inflammatory cytokines IL-10 and IL-1RA. Data are shown as mean ± SEM.
    Middlebrook 7h10 Agar, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 490 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 agar
    The MIC distribution of meropenem-clavulanic acid for the M. tuberculosis isolates ( n = 68) tested using <t>Middlebrook</t> <t>7H10</t> medium, shown by bars shaded as follows: non-MDR/XDR-TB (gray); XDR-TB (black), and MDR-TB (hatched). The MIC distribution shows
    Middlebrook 7h10 Agar, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 96/100, based on 319 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals middlebrook 7h10 agar
    Effect of inhibition of ROS and NO production on the survival of Mtb strains inside MØs. MØs were incubated with apocynin or L-NIL for 30 minutes, then infected with Mtb wild-type or Δ( ku,ligD,recA ) for 2 hours (MOI = 10) and washed with HBSS. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto <t>Middlebrook</t> <t>7H10</t> agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; #p≤0.04, Δ( ku,ligD,recA ) vs. Δ( ku,ligD,recA ) + apocynin or L-NIL; Mann-Whitney U test).
    Middlebrook 7h10 Agar, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 99/100, based on 3002 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Hardy Diagnostics middlebrook 7h10 plates
    Effect of inhibition of ROS and NO production on the survival of Mtb strains inside MØs. MØs were incubated with apocynin or L-NIL for 30 minutes, then infected with Mtb wild-type or Δ( ku,ligD,recA ) for 2 hours (MOI = 10) and washed with HBSS. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto <t>Middlebrook</t> <t>7H10</t> agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; #p≤0.04, Δ( ku,ligD,recA ) vs. Δ( ku,ligD,recA ) + apocynin or L-NIL; Mann-Whitney U test).
    Middlebrook 7h10 Plates, supplied by Hardy Diagnostics, used in various techniques. Bioz Stars score: 94/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco middlebrook 7h10 plates
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Plates, supplied by Difco, used in various techniques. Bioz Stars score: 90/100, based on 37 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Toxicology Inc middlebrook 7h10 agar
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Agar, supplied by Molecular Toxicology Inc, used in various techniques. Bioz Stars score: 91/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HiMedia Laboratories middlebrook 7h10 agar
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Agar, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 90/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 plates
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Plates, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 92/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 oadc
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Oadc, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Hardy Diagnostics middlebrook 7h10 agar
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Agar, supplied by Hardy Diagnostics, used in various techniques. Bioz Stars score: 88/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 sel7h11 agar
    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in <t>Middlebrook</t> <t>7H10</t> agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P
    Middlebrook 7h10 Sel7h11 Agar, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco middlebrook 7h10 medium
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 Medium, supplied by Difco, used in various techniques. Bioz Stars score: 93/100, based on 152 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 medium
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Difco middlebrook 7h10 bacto agar
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 Bacto Agar, supplied by Difco, used in various techniques. Bioz Stars score: 81/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson middlebrook 7h10 s7h11 biplate
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 S7h11 Biplate, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 86/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Merck & Co middlebrook 7h10
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10, supplied by Merck & Co, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10, supplied by Fisher Scientific, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 Medium, supplied by HiMedia Laboratories, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals middlebrook 7h10 11
    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on <t>Middlebrook</t> <t>7H10</t> agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P
    Middlebrook 7h10 11, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 94/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on <t>7H10</t> agar plates or <t>7H9/OADC/Tween</t> broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P
    Middlebrook 7h10 Oadc, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 33 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on <t>7H10</t> agar plates or <t>7H9/OADC/Tween</t> broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P
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    In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on <t>7H10</t> agar plates or <t>7H9/OADC/Tween</t> broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P
    Middlebrook 7h10 Broth, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 41 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on <t>7H10</t> agar plates or <t>7H9/OADC/Tween</t> broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P
    Middlebrook 7h10 Powder, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 85/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals middlebrook 7h10 plates
    Growth of M. bovis Δ nadABC and M. tuberculosis Δ nadABC in vitro. A . The strains were grown with NAm (20 mg/l) to log phase, spun down, washed 5 X with PBS, resuspended in media without NAm, and inoculated into media containing either NAm at different concentrations or NAc at 20 mg/l. The complemented strains were grown without any supplement. The cultures were incubated with shaking at 37°C for 3 weeks and growth was followed by measuring OD 600nm . B . The strains were grown, washed, diluted, and inoculated in media containing or not containing NAm (20 mg/l). At each time point, samples were taken, diluted, and plated onto <t>Middlebrook</t> <t>7H10</t> plates containing OADC, glycerol, and NAm (20 mg/l). The plates were incubated at 37°C for 4-5 weeks and colonies were counted. The concentrations are in mg/l.
    Middlebrook 7h10 Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 56 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    garA is required for growth of M . tuberculosis in vitro , survival in macrophages, and virulence in mice. ( A ) M . tuberculosis lacking garA was unable to grow on standard 7H10 medium unless supplemented with asparagine. Plasmid-borne garA restored the defect, but variants of garA with mutations at threonine 21 in the phosphorylation motif (ETTS) gave only partial complementation. Strains were grown in Middlebrook 7H9 plus 30 mM asparagine then washed and diluted in standard 7H9 and spotted onto standard 7H10 with or without 10 mM asparagine. Photographs are representative of at least 3 independent experiments. ( B ) M . tuberculosis lacking garA (red squares) had a defect in growth and survival in differentiated THP-1 cells compared to parental M . tuberculosis H37Rv (black circles). Re-introduction of GarA (black triangles) or variants of GarA lacking a single phosphorylation site (grey crosses and squares) restored growth but variant GarA lacking both phosphorylation sites (green triangles) did not. Data points show the mean and standard deviation from four replicates and are representative of two independent experiments. ( C ) M . tuberculosis lacking garA was avirulent in mice as it was eliminated from the lungs. BALB/C mice were infected intranasally with 10 5 bacilli and bacterial burden was measured on days 1, 7, 21 and 28. Data points show the bacterial burden in individual animals. The bacterial burden of mice infected with Δ garA Mt (red squares) was significantly lower than those infected with M . tuberculosis H37Rv (black circles), or complemented Δ garA Mt (black triangles) at all time points from day 7 (p

    Journal: PLoS Pathogens

    Article Title: PknG senses amino acid availability to control metabolism and virulence of Mycobacterium tuberculosis

    doi: 10.1371/journal.ppat.1006399

    Figure Lengend Snippet: garA is required for growth of M . tuberculosis in vitro , survival in macrophages, and virulence in mice. ( A ) M . tuberculosis lacking garA was unable to grow on standard 7H10 medium unless supplemented with asparagine. Plasmid-borne garA restored the defect, but variants of garA with mutations at threonine 21 in the phosphorylation motif (ETTS) gave only partial complementation. Strains were grown in Middlebrook 7H9 plus 30 mM asparagine then washed and diluted in standard 7H9 and spotted onto standard 7H10 with or without 10 mM asparagine. Photographs are representative of at least 3 independent experiments. ( B ) M . tuberculosis lacking garA (red squares) had a defect in growth and survival in differentiated THP-1 cells compared to parental M . tuberculosis H37Rv (black circles). Re-introduction of GarA (black triangles) or variants of GarA lacking a single phosphorylation site (grey crosses and squares) restored growth but variant GarA lacking both phosphorylation sites (green triangles) did not. Data points show the mean and standard deviation from four replicates and are representative of two independent experiments. ( C ) M . tuberculosis lacking garA was avirulent in mice as it was eliminated from the lungs. BALB/C mice were infected intranasally with 10 5 bacilli and bacterial burden was measured on days 1, 7, 21 and 28. Data points show the bacterial burden in individual animals. The bacterial burden of mice infected with Δ garA Mt (red squares) was significantly lower than those infected with M . tuberculosis H37Rv (black circles), or complemented Δ garA Mt (black triangles) at all time points from day 7 (p

    Article Snippet: Bacterial strains, media, and culture M . tuberculosis H37Rv and M . smegmatis mc2 155 were routinely cultured on Middlebrook 7H10 agar (Oxoid) with 10% ADN (0.5% bovine serum albumin, 0.2% dextrose, 0.085% NaCl) and Middlebrook 7H9 medium (Oxoid) with 10% ADN and 0.05% Tween 80.

    Techniques: In Vitro, Mouse Assay, Plasmid Preparation, Variant Assay, Standard Deviation, Infection

    Growth of M. smegmatis mc 2 155 and M371 under fluoroquinolones exposure. Ten-fold serial dilutions of wild-type, M371, M371-pALACE- pafC and M371-pALACE were spotted on Middlebrook 7H10 containing indicated concentration of moxifloxacin, norfloxacin, ofloxacin and ciprofloxacin. Then the result was recorded when incubated at 37 °C for 3 days.

    Journal: Scientific Reports

    Article Title: Proteasome Accessory Factor C (pafC) Is a novel gene Involved in Mycobacterium Intrinsic Resistance to broad-spectrum antibiotics - Fluoroquinolones

    doi: 10.1038/srep11910

    Figure Lengend Snippet: Growth of M. smegmatis mc 2 155 and M371 under fluoroquinolones exposure. Ten-fold serial dilutions of wild-type, M371, M371-pALACE- pafC and M371-pALACE were spotted on Middlebrook 7H10 containing indicated concentration of moxifloxacin, norfloxacin, ofloxacin and ciprofloxacin. Then the result was recorded when incubated at 37 °C for 3 days.

    Article Snippet: Bacterial strains, plasmid, and growth conditions M. smegmatis mc2 155 and M371 were grown in Middlebrook 7H9 medium supplemented with 0.05% Tween80 and 0.2% glycerinum or Middlebrook 7H10 plates supplemented with 0.5% glycerinum.

    Techniques: Concentration Assay, Incubation

    Modified Middlebrook 7H10 medium containing 250 mg/L MG with mycobacteria primarily isolated from soil. a Sample collected at campus, b sample collected at community, and c collected at natural park. Black arrows indicate mycobacterial colonies, red arrows indicate dried soils

    Journal: AMB Express

    Article Title: Isolation of nontuberculous mycobacteria from soil using Middlebrook 7H10 agar with increased malachite green concentration

    doi: 10.1186/s13568-017-0373-6

    Figure Lengend Snippet: Modified Middlebrook 7H10 medium containing 250 mg/L MG with mycobacteria primarily isolated from soil. a Sample collected at campus, b sample collected at community, and c collected at natural park. Black arrows indicate mycobacterial colonies, red arrows indicate dried soils

    Article Snippet: Thus, modified Middlebrook 7H10 media with MG at concentrations of 100–250 mg/L were found to be promising for culturing NTM from soil.

    Techniques: Modification, Isolation

    mc 2 7901 and mc 2 7902 grow similarly to virulent M. tuberculosis in vitro . (A) Log-phase cultures of mc 2 7901 and mc 2 7902 grown in Middlebrook 7H9-OADC-glycerol-tyloxapol-PLAM were diluted 1/100, and growth was followed by recording optical density at 600 nm (OD 600 ) over time. Mean with standard deviation is plotted ( n = 2). (B) RAW 264.7 macrophages were infected at an MOI of 1 with mc 2 7901, mc 2 7902, or H37Rv. At the indicated time points, macrophages were lysed, and bacterial titers were determined by plating for CFU on Middlebrook 7H10-OADC-glycerol-PLAM plates. PLAM was added to the macrophage growth medium, and the medium was changed at each time point. (C) Growth of mc 2 7901 and mc 2 7902 in RAW 264.7 macrophages relative to the inocula (same experiment as in panel B). Mean with standard deviation is plotted ( n = 2).

    Journal: mBio

    Article Title: Rational Design of Biosafety Level 2-Approved, Multidrug-Resistant Strains of Mycobacterium tuberculosis through Nutrient Auxotrophy

    doi: 10.1128/mBio.00938-18

    Figure Lengend Snippet: mc 2 7901 and mc 2 7902 grow similarly to virulent M. tuberculosis in vitro . (A) Log-phase cultures of mc 2 7901 and mc 2 7902 grown in Middlebrook 7H9-OADC-glycerol-tyloxapol-PLAM were diluted 1/100, and growth was followed by recording optical density at 600 nm (OD 600 ) over time. Mean with standard deviation is plotted ( n = 2). (B) RAW 264.7 macrophages were infected at an MOI of 1 with mc 2 7901, mc 2 7902, or H37Rv. At the indicated time points, macrophages were lysed, and bacterial titers were determined by plating for CFU on Middlebrook 7H10-OADC-glycerol-PLAM plates. PLAM was added to the macrophage growth medium, and the medium was changed at each time point. (C) Growth of mc 2 7901 and mc 2 7902 in RAW 264.7 macrophages relative to the inocula (same experiment as in panel B). Mean with standard deviation is plotted ( n = 2).

    Article Snippet: Middlebrook 7H10 (Difco) supplemented with 10% (vol/vol) OADC and 0.2% (vol/vol) glycerol was used as solid medium.

    Techniques: In Vitro, Standard Deviation, Infection

    mc 2 7901- and mc 2 7902-derived MDR strains grow slower than parental strains in vitro and are killed by second-line TB drugs or nutrient starvation. (A) Log-phase cultures were diluted 1/100, and growth was followed by recording optical density at 600 nm. Mean with standard deviation is plotted ( n = 2). (B) RAW 264.7 macrophages were infected at an MOI of 1. At the indicated time points, macrophages were lysed to determine bacterial titers. PLAM was added to the macrophage growth medium, and the medium was changed at each time point. (C) PLAM-supplemented log-phase cultures of mc 2 7901, mc 2 7902, mc 2 8248, and mc 2 8251 were treated with EOK (ethionamide [25 mg/liter], ofloxacin [5 mg/liter], kanamycin [20 mg/liter]). (D) Log-phase cultures of mc 2 7901, mc 2 7902, mc 2 8248, and mc 2 8251 were washed five times in PBS and resuspended in Middlebrook 7H9-OADC-glycerol-tyloxapol containing PLAM (dilution factor 1/100) or not. In experiments in panels B, C, and D, the strains were initially grown in Middlebrook 7H9-OADC-glycerol-tyloxapol-PLAM. Samples were taken at the indicated time points, diluted, and plated for CFU on Middlebrook 7H10-OADC-glycerol-PLAM plates. Means with standard deviations are plotted ( n = 3).

    Journal: mBio

    Article Title: Rational Design of Biosafety Level 2-Approved, Multidrug-Resistant Strains of Mycobacterium tuberculosis through Nutrient Auxotrophy

    doi: 10.1128/mBio.00938-18

    Figure Lengend Snippet: mc 2 7901- and mc 2 7902-derived MDR strains grow slower than parental strains in vitro and are killed by second-line TB drugs or nutrient starvation. (A) Log-phase cultures were diluted 1/100, and growth was followed by recording optical density at 600 nm. Mean with standard deviation is plotted ( n = 2). (B) RAW 264.7 macrophages were infected at an MOI of 1. At the indicated time points, macrophages were lysed to determine bacterial titers. PLAM was added to the macrophage growth medium, and the medium was changed at each time point. (C) PLAM-supplemented log-phase cultures of mc 2 7901, mc 2 7902, mc 2 8248, and mc 2 8251 were treated with EOK (ethionamide [25 mg/liter], ofloxacin [5 mg/liter], kanamycin [20 mg/liter]). (D) Log-phase cultures of mc 2 7901, mc 2 7902, mc 2 8248, and mc 2 8251 were washed five times in PBS and resuspended in Middlebrook 7H9-OADC-glycerol-tyloxapol containing PLAM (dilution factor 1/100) or not. In experiments in panels B, C, and D, the strains were initially grown in Middlebrook 7H9-OADC-glycerol-tyloxapol-PLAM. Samples were taken at the indicated time points, diluted, and plated for CFU on Middlebrook 7H10-OADC-glycerol-PLAM plates. Means with standard deviations are plotted ( n = 3).

    Article Snippet: Middlebrook 7H10 (Difco) supplemented with 10% (vol/vol) OADC and 0.2% (vol/vol) glycerol was used as solid medium.

    Techniques: Derivative Assay, In Vitro, Standard Deviation, Infection

    Bacterial loads in spleens (A) and livers (B) of C3-sufficient and C3-deficient mice were similar after 1 or 5 weeks of infection. C3-sufficient and C3-deficient mice with both resistant (bcg-r) and susceptible (bcg-s) backgrounds were retroorbitally infected with 10 6 M. avium 724 cells, and the mice were maintained for 1 or 5 weeks. The mice were sacrificed, spleens and livers were homogenized, and serial dilutions were plated onto Middlebrook 7H10 agar to determine the numbers of CFU per organ. The data are representative of at least two experiments, and each data point represents the data from four to seven mice; the error bars indicate standard deviations. wt, wild type; ko, C3 knockout.

    Journal: Infection and Immunity

    Article Title: Role of Complement in Mycobacterium avium Pathogenesis: In Vivo and In Vitro Analyses of the Host Response to Infection in the Absence of Complement Component C3

    doi: 10.1128/IAI.69.12.7729-7735.2001

    Figure Lengend Snippet: Bacterial loads in spleens (A) and livers (B) of C3-sufficient and C3-deficient mice were similar after 1 or 5 weeks of infection. C3-sufficient and C3-deficient mice with both resistant (bcg-r) and susceptible (bcg-s) backgrounds were retroorbitally infected with 10 6 M. avium 724 cells, and the mice were maintained for 1 or 5 weeks. The mice were sacrificed, spleens and livers were homogenized, and serial dilutions were plated onto Middlebrook 7H10 agar to determine the numbers of CFU per organ. The data are representative of at least two experiments, and each data point represents the data from four to seven mice; the error bars indicate standard deviations. wt, wild type; ko, C3 knockout.

    Article Snippet: To generate M. avium stocks, bacteria were passaged through a mouse to ensure virulence, and a single colony was used to inoculate Middlebrook 7H10 media (Difco, Detroit, Mich.) supplemented with glucose, oleic acid, albumin, Tween 20, and NaCl (supplemented Middlebrook media).

    Techniques: Mouse Assay, Infection, Knock-Out

    Survival of M. tuberculosis ( Mtb ) inside human macrophages. Human MDMs infected with Mtb H37Rv (MOI 5). The percentages of intracellular and not phagocytosed bacilli immediately after infection were determined by CFU assay and compared to inoculum (a). Lysed infected macrophages and released bacteria in the culture supernatants at 1, 3, 7 and 11 days after infection were plated on Middlebrook 7H10 with OADC and the CFU were counted after 14 days (b). Cytotoxicity induced by Mtb in human MDMs was measured by LDH activity (c). Values are the means ± SD from four independent experiments. The percentage of infected cells was measured, after 3 hr of infection (time 0) and 1, 3, 7 days after infection, by acridine-orange staining (d). and the representative pictures of infected MDMs, are shown (e). A significant difference was detected with respect to 1 and 3 days post-infection (* P

    Journal: Immunology

    Article Title: Gene expression profiling of human macrophages at late time of infection with Mycobacterium tuberculosis

    doi: 10.1111/j.1365-2567.2006.02378.x

    Figure Lengend Snippet: Survival of M. tuberculosis ( Mtb ) inside human macrophages. Human MDMs infected with Mtb H37Rv (MOI 5). The percentages of intracellular and not phagocytosed bacilli immediately after infection were determined by CFU assay and compared to inoculum (a). Lysed infected macrophages and released bacteria in the culture supernatants at 1, 3, 7 and 11 days after infection were plated on Middlebrook 7H10 with OADC and the CFU were counted after 14 days (b). Cytotoxicity induced by Mtb in human MDMs was measured by LDH activity (c). Values are the means ± SD from four independent experiments. The percentage of infected cells was measured, after 3 hr of infection (time 0) and 1, 3, 7 days after infection, by acridine-orange staining (d). and the representative pictures of infected MDMs, are shown (e). A significant difference was detected with respect to 1 and 3 days post-infection (* P

    Article Snippet: Representative samples were thawed and colony-forming units (CFUs) per ml were enumerated by plating on Middlebrook 7H10 (Becton Dickinson, Cockeysville, MD) supplemented with Middlebrook oleic acid-albumin-dextrose-catalase (OADC).

    Techniques: Infection, Colony-forming Unit Assay, Activity Assay, Staining

    Effects of euglycaemic and hyperglycaemic culture conditions on H37RV infection and cytokine production in vitro. (A) CD14 + selected monocytes were differentiated into M2 macrophages in the presence of 5 mmol/L glucose, 5 mmol/L glucose and 20 mmol/L mannitol or 25 mmol/L glucose. The macrophages were infected for 1 hour with H37Rv at an MOI of 10. After infection the macrophages were washed and fresh media containing the different glucose media was added. After 24 hours supernatants were collected and the cells were lysed by osmotic pressure. (B) Cell lysates were serially diluted and plated on Middlebrook 7H10 agar. CFUs were counted after 2–3 weeks of growth at 37°C. Data are shown as mean ± SEM of two independent experiments (n = 4). (C) Pro-inflammatory cytokines TNF-α and IL-6 were measured along with the anti-inflammatory cytokines IL-10 and IL-1RA. Data are shown as mean ± SEM.

    Journal: PLoS ONE

    Article Title: The Effect of Hyperglycaemia on In Vitro Cytokine Production and Macrophage Infection with Mycobacterium tuberculosis

    doi: 10.1371/journal.pone.0117941

    Figure Lengend Snippet: Effects of euglycaemic and hyperglycaemic culture conditions on H37RV infection and cytokine production in vitro. (A) CD14 + selected monocytes were differentiated into M2 macrophages in the presence of 5 mmol/L glucose, 5 mmol/L glucose and 20 mmol/L mannitol or 25 mmol/L glucose. The macrophages were infected for 1 hour with H37Rv at an MOI of 10. After infection the macrophages were washed and fresh media containing the different glucose media was added. After 24 hours supernatants were collected and the cells were lysed by osmotic pressure. (B) Cell lysates were serially diluted and plated on Middlebrook 7H10 agar. CFUs were counted after 2–3 weeks of growth at 37°C. Data are shown as mean ± SEM of two independent experiments (n = 4). (C) Pro-inflammatory cytokines TNF-α and IL-6 were measured along with the anti-inflammatory cytokines IL-10 and IL-1RA. Data are shown as mean ± SEM.

    Article Snippet: M2 macrophages were lysed in water for 5 minutes and plated on Middlebrook 7H10 agar (Difco, Becton-Dickinson) supplemented with OADC.

    Techniques: Infection, In Vitro

    The MIC distribution of meropenem-clavulanic acid for the M. tuberculosis isolates ( n = 68) tested using Middlebrook 7H10 medium, shown by bars shaded as follows: non-MDR/XDR-TB (gray); XDR-TB (black), and MDR-TB (hatched). The MIC distribution shows

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Meropenem-Clavulanic Acid Has High In Vitro Activity against Multidrug-Resistant Mycobacterium tuberculosis

    doi: 10.1128/AAC.00171-15

    Figure Lengend Snippet: The MIC distribution of meropenem-clavulanic acid for the M. tuberculosis isolates ( n = 68) tested using Middlebrook 7H10 medium, shown by bars shaded as follows: non-MDR/XDR-TB (gray); XDR-TB (black), and MDR-TB (hatched). The MIC distribution shows

    Article Snippet: Middlebrook 7H10 agar (Becton Dickinson AB, Stockholm, Sweden) enriched with 10% oleic acid–albumin–dextrose–catalase (OADC) and 5% glycerol was prepared in 14-cm petri dishes, each dish containing 60 ml of agar.

    Techniques:

    Effect of inhibition of ROS and NO production on the survival of Mtb strains inside MØs. MØs were incubated with apocynin or L-NIL for 30 minutes, then infected with Mtb wild-type or Δ( ku,ligD,recA ) for 2 hours (MOI = 10) and washed with HBSS. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto Middlebrook 7H10 agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; #p≤0.04, Δ( ku,ligD,recA ) vs. Δ( ku,ligD,recA ) + apocynin or L-NIL; Mann-Whitney U test).

    Journal: PLoS ONE

    Article Title: Either Non-Homologous Ends Joining or Homologous Recombination Is Required to Repair Double-Strand Breaks in the Genome of Macrophage-Internalized Mycobacterium tuberculosis

    doi: 10.1371/journal.pone.0092799

    Figure Lengend Snippet: Effect of inhibition of ROS and NO production on the survival of Mtb strains inside MØs. MØs were incubated with apocynin or L-NIL for 30 minutes, then infected with Mtb wild-type or Δ( ku,ligD,recA ) for 2 hours (MOI = 10) and washed with HBSS. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto Middlebrook 7H10 agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; #p≤0.04, Δ( ku,ligD,recA ) vs. Δ( ku,ligD,recA ) + apocynin or L-NIL; Mann-Whitney U test).

    Article Snippet: A sample of each strain was taken during exponential and stationary phases of Mtb growth, and serial dilutions were prepared and plated on Middlebrook 7H10 agar supplemented with 10% OADC.

    Techniques: Inhibition, Incubation, Infection, MANN-WHITNEY

    Survival of Mtb strains inside MØs. (A) MØs were infected with Mtb wild-type or its mutants, Δ( ku,ligD,recA ), Δ( ku,ligD ) or Δ recA , for 2 hours (MOI = 10), washed with HBSS, and cultured for 2, 4, and 6 days. (B and C) MØs were infected with Mtb wild-type or its mutants (B) or complemented strains Δ( ku,ligD,recA )- P recA recA – complemented strain I or Δ( ku,ligD,recA )- P kuligD ku-ligD – complemented strain II (C) for 6 days. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto Middlebrook 7H10 agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; *p≤0.05, Δ( ku,ligD,recA ) vs. wild-type; #p≤0.05, complemented strain I and II vs. Δ( ku,ligD,recA ) Mann-Whitney U test).

    Journal: PLoS ONE

    Article Title: Either Non-Homologous Ends Joining or Homologous Recombination Is Required to Repair Double-Strand Breaks in the Genome of Macrophage-Internalized Mycobacterium tuberculosis

    doi: 10.1371/journal.pone.0092799

    Figure Lengend Snippet: Survival of Mtb strains inside MØs. (A) MØs were infected with Mtb wild-type or its mutants, Δ( ku,ligD,recA ), Δ( ku,ligD ) or Δ recA , for 2 hours (MOI = 10), washed with HBSS, and cultured for 2, 4, and 6 days. (B and C) MØs were infected with Mtb wild-type or its mutants (B) or complemented strains Δ( ku,ligD,recA )- P recA recA – complemented strain I or Δ( ku,ligD,recA )- P kuligD ku-ligD – complemented strain II (C) for 6 days. On the day of infection and after 6 days of culture, MØs were lysed with Triton X-100 and cell lysates were plated onto Middlebrook 7H10 agar supplemented with 10% OADC. After 21 days of culture, CFUs were counted. Data are presented as fold increase in CFUs, expressed as means ± SEM (n = 5; *p≤0.05, Δ( ku,ligD,recA ) vs. wild-type; #p≤0.05, complemented strain I and II vs. Δ( ku,ligD,recA ) Mann-Whitney U test).

    Article Snippet: A sample of each strain was taken during exponential and stationary phases of Mtb growth, and serial dilutions were prepared and plated on Middlebrook 7H10 agar supplemented with 10% OADC.

    Techniques: Infection, Cell Culture, MANN-WHITNEY

    MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in Middlebrook 7H10 agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Multidrug Resistance of a Porin Deletion Mutant of Mycobacterium smegmatis

    doi: 10.1128/AAC.48.11.4163-4170.2004

    Figure Lengend Snippet: MspA-dependent sensitivities and permeabilities of M. smegmatis to hydrophobic compounds. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to erythromycin were determined by serial dilution in Middlebrook 7H10 agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). (B) The accumulation of [ 14 C]chenodeoxycholate by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. Regression analysis yielded uptake rates of 0.09 and 0.03 nmol mg −1 min −1 for 20 μM chenodeoxycholate for the wild-type and mutant strains of M. smegmatis , respectively. The assay was performed at 37°C. The structures of the compounds are shown in the corresponding graphs. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P

    Article Snippet: All mycobacterial strains were grown in Middlebrook 7H9 liquid medium (Difco) supplemented with 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H10 plates (Difco) containing 0.2% glycerol at 37°C.

    Techniques: Mutagenesis, Transformation Assay, Expressing, Plasmid Preparation, Serial Dilution, Concentration Assay

    MspA-dependent sensitivity and permeability of M. smegmatis to isoniazid. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to isoniazid were determined by serial dilution in Middlebrook 7H10 agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). The structure of isoniazid is shown. (B) The accumulation of 20 μM [ 14 C]isoniazid by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. The assay was performed at 37°C. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Multidrug Resistance of a Porin Deletion Mutant of Mycobacterium smegmatis

    doi: 10.1128/AAC.48.11.4163-4170.2004

    Figure Lengend Snippet: MspA-dependent sensitivity and permeability of M. smegmatis to isoniazid. (A) The sensitivities of wild-type M. smegmatis (black bars), the Δ mspA mutant (white bars), and the Δ mspA mutant transformed with the mspA expression vector pMN014 (gray bars) to isoniazid were determined by serial dilution in Middlebrook 7H10 agar (c, concentration). The number of surviving cells was normalized to the number of cells counted on plates without antibiotic for each strain, and the results are expressed as relative CFU (percent CFU). The structure of isoniazid is shown. (B) The accumulation of 20 μM [ 14 C]isoniazid by M. smegmatis SMR5 (wild type; closed circles) and M. smegmatis MN01 (Δ mspA ; open circles) was measured. The assay was performed at 37°C. The sensitivity and uptake experiments were done in triplicate, and the results are shown with their standard deviations. The asterisks denote the datum points that differed significantly ( P

    Article Snippet: All mycobacterial strains were grown in Middlebrook 7H9 liquid medium (Difco) supplemented with 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H10 plates (Difco) containing 0.2% glycerol at 37°C.

    Techniques: Permeability, Mutagenesis, Transformation Assay, Expressing, Plasmid Preparation, Serial Dilution, Concentration Assay

    BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on Middlebrook 7H10 agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P

    Journal: BMC Immunology

    Article Title: Rapid loss of early antigen-presenting activity of lymph node dendritic cells against Ag85A protein following Mycobacterium bovis BCG infection

    doi: 10.1186/s12865-018-0258-8

    Figure Lengend Snippet: BCG infection kinetics of murine LN DCs. Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU rBCG-GFP, and LN cells were harvested at different time points. DCs were sorted and analyzed for the presence of rBCG-GFP. Infection of murine LN DCs with BCG ( a ). Six groups of mice ( n = 6) were s.c. injected with 1 × 10 8 CFU BCG, and BCG in DCs was quantified in CFUs by culturing on Middlebrook 7H10 agar ( b ). The results are representative of three independent experiments and presented as means ± SEM. Statistical significance was determined using Student’s t -test (* P

    Article Snippet: Both strains were grown with gentle agitation (80 rpm) in Middlebrook 7H9 medium (Difco, Detroit, MI, USA) supplemented with 0.05% Tween 80 and 10% albumin-dextrose-catalase (ADC) enrichment or on solid Middlebrook 7H10 medium (Difco) supplemented with 0.05% Tween 80 and 10% oleic-ADC enrichment.

    Techniques: Infection, Mouse Assay, Injection

    In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on 7H10 agar plates or 7H9/OADC/Tween broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Mycobacterium abscessus cording prevents phagocytosis and promotes abscess formation

    doi: 10.1073/pnas.1321390111

    Figure Lengend Snippet: In vivo cording of the R variant initiates abscess formation. ( A ) Fluorescent images of R or S morphotypes on 7H10 agar plates or 7H9/OADC/Tween broth. The S variant appears hemispherical and smooth, whereas the R variant displays serpentine cords. Only rough bacteria produce multicellular cords. (Scale bars, 1 mm.) ( B and C ) R-variant–infected embryos expressing mCherry ( B ) or GFP ( C ) at 2 dpi. ( B ) Fluorescence and DIC overlay of the head. (Scale bar, 150 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of a serpentine cord. ( C ) Fluorescence and DIC overlay of part of the tail. (Scale bar, 100 μm.) The enlarged view at right shows a maximum intensity projection of confocal imaging of cords within the spinal cord. ( D ) Kinetic of cord formation in infected embryos (∼300 cfu of M. abscessus S or R variant; n = 40). Only embryos infected with the R variant develop serpentine cords ( P

    Article Snippet: Several dilutions of homogenates were plated on Middlebrook 7H10/OADC and supplemented with BBL MGIT PANTA (BD) as recommended by the supplier.

    Techniques: In Vivo, Variant Assay, Infection, Expressing, Fluorescence, Imaging

    Loss of mmpL4b confers high pathogenicity to the S variant. ( A ) Fluorescent pictures showing the general aspect of the S variant M. abscessus Δ mmpL4b mutant (SΔ mmpL4b ) and its complemented strain (SΔ mmpL4b _C) on 7H10 agar or in 7H9/OADC/Tween broth medium. Hemispherical, smooth colonies are characteristic of SΔ mmpL4b_ C, whereas the SΔ mmpL4b strain forms serpentine cords. (Scale bars, 0.5 mm.) ( B ) Fluorescence and DIC overlay of the whole head of infected embryos with red fluorescent SΔ mmpL4b showing large numbers of serpentine cord within the brain. ( C ) Survival of embryos infected with ∼200 cfu M. abscessus R variant, S variant, SΔ mmpL4b , or SΔ mmpL4b_ C ( n = 30–40 per group). Shown are representative results from two independent experiments. Embryos are significantly more susceptible to infection by the SΔ mmpL4b mutant and the R variant than by the parental M. abscessus S variant ( P

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Mycobacterium abscessus cording prevents phagocytosis and promotes abscess formation

    doi: 10.1073/pnas.1321390111

    Figure Lengend Snippet: Loss of mmpL4b confers high pathogenicity to the S variant. ( A ) Fluorescent pictures showing the general aspect of the S variant M. abscessus Δ mmpL4b mutant (SΔ mmpL4b ) and its complemented strain (SΔ mmpL4b _C) on 7H10 agar or in 7H9/OADC/Tween broth medium. Hemispherical, smooth colonies are characteristic of SΔ mmpL4b_ C, whereas the SΔ mmpL4b strain forms serpentine cords. (Scale bars, 0.5 mm.) ( B ) Fluorescence and DIC overlay of the whole head of infected embryos with red fluorescent SΔ mmpL4b showing large numbers of serpentine cord within the brain. ( C ) Survival of embryos infected with ∼200 cfu M. abscessus R variant, S variant, SΔ mmpL4b , or SΔ mmpL4b_ C ( n = 30–40 per group). Shown are representative results from two independent experiments. Embryos are significantly more susceptible to infection by the SΔ mmpL4b mutant and the R variant than by the parental M. abscessus S variant ( P

    Article Snippet: Several dilutions of homogenates were plated on Middlebrook 7H10/OADC and supplemented with BBL MGIT PANTA (BD) as recommended by the supplier.

    Techniques: Variant Assay, Mutagenesis, Fluorescence, Infection

    Growth of M. bovis Δ nadABC and M. tuberculosis Δ nadABC in vitro. A . The strains were grown with NAm (20 mg/l) to log phase, spun down, washed 5 X with PBS, resuspended in media without NAm, and inoculated into media containing either NAm at different concentrations or NAc at 20 mg/l. The complemented strains were grown without any supplement. The cultures were incubated with shaking at 37°C for 3 weeks and growth was followed by measuring OD 600nm . B . The strains were grown, washed, diluted, and inoculated in media containing or not containing NAm (20 mg/l). At each time point, samples were taken, diluted, and plated onto Middlebrook 7H10 plates containing OADC, glycerol, and NAm (20 mg/l). The plates were incubated at 37°C for 4-5 weeks and colonies were counted. The concentrations are in mg/l.

    Journal: Molecular microbiology

    Article Title: NAD+ Auxotrophy is Bacteriocidal for the Tubercle Bacilli

    doi: 10.1111/j.1365-2958.2010.07099.x

    Figure Lengend Snippet: Growth of M. bovis Δ nadABC and M. tuberculosis Δ nadABC in vitro. A . The strains were grown with NAm (20 mg/l) to log phase, spun down, washed 5 X with PBS, resuspended in media without NAm, and inoculated into media containing either NAm at different concentrations or NAc at 20 mg/l. The complemented strains were grown without any supplement. The cultures were incubated with shaking at 37°C for 3 weeks and growth was followed by measuring OD 600nm . B . The strains were grown, washed, diluted, and inoculated in media containing or not containing NAm (20 mg/l). At each time point, samples were taken, diluted, and plated onto Middlebrook 7H10 plates containing OADC, glycerol, and NAm (20 mg/l). The plates were incubated at 37°C for 4-5 weeks and colonies were counted. The concentrations are in mg/l.

    Article Snippet: The lysates were diluted and plated on Middlebrook 7H10 plates (see above) containing NAm if necessary.

    Techniques: In Vitro, Incubation

    Growth of M. bovis Δ nadABC pMV261:: pncA TB in vitro. M. bovis Δ nadABC was transformed with M. tuberculosis pncA cloned into the replicative plasmid pMV261. A . M. bovis Δ nadABC pMV261:: pncA TB and its parent strain M. bovis Δ nadABC were grown in NAm-containing media, washed 5 X in PBS, diluted, and inoculated in media containing NAm (20 and 0.1 mg/l). Growth was followed by measuring OD 600nm at diverse time points. B . M. bovis Δ nadABC pMV261:: pncA TB and M. bovis Δ nadABC were grown and washed as described above, diluted 1-50, and inoculated in media without NAm (20 mg/l). At each time, samples were taken, diluted, and plated onto Middlebrook 7H10 plates containing NAm (20 mg/l). The plates were incubated at 37°C for 4-5 weeks and colonies were counted.

    Journal: Molecular microbiology

    Article Title: NAD+ Auxotrophy is Bacteriocidal for the Tubercle Bacilli

    doi: 10.1111/j.1365-2958.2010.07099.x

    Figure Lengend Snippet: Growth of M. bovis Δ nadABC pMV261:: pncA TB in vitro. M. bovis Δ nadABC was transformed with M. tuberculosis pncA cloned into the replicative plasmid pMV261. A . M. bovis Δ nadABC pMV261:: pncA TB and its parent strain M. bovis Δ nadABC were grown in NAm-containing media, washed 5 X in PBS, diluted, and inoculated in media containing NAm (20 and 0.1 mg/l). Growth was followed by measuring OD 600nm at diverse time points. B . M. bovis Δ nadABC pMV261:: pncA TB and M. bovis Δ nadABC were grown and washed as described above, diluted 1-50, and inoculated in media without NAm (20 mg/l). At each time, samples were taken, diluted, and plated onto Middlebrook 7H10 plates containing NAm (20 mg/l). The plates were incubated at 37°C for 4-5 weeks and colonies were counted.

    Article Snippet: The lysates were diluted and plated on Middlebrook 7H10 plates (see above) containing NAm if necessary.

    Techniques: In Vitro, Transformation Assay, Clone Assay, Plasmid Preparation, Incubation