middlebrook 7h10 plates Search Results


  • Logo
  • About
  • News
  • Press Release
  • Team
  • Advisors
  • Partners
  • Contact
  • Bioz Stars
  • Bioz vStars
  • 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
    https://www.bioz.com/result/middlebrook 7h10 agar/product/Millipore
    Average 99 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 agar - by Bioz Stars, 2020-04
    99/100 stars
      Buy from Supplier

    85
    Thermo Fisher middlebrook 7h10
    Middlebrook 7h10, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 85/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10/product/Thermo Fisher
    Average 85 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 - by Bioz Stars, 2020-04
    85/100 stars
      Buy from Supplier

    94
    Hardy Diagnostics middlebrook 7h10 plates
    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
    https://www.bioz.com/result/middlebrook 7h10 plates/product/Hardy Diagnostics
    Average 94 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 plates - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    90
    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
    https://www.bioz.com/result/middlebrook 7h10 plates/product/Difco
    Average 90 stars, based on 37 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 plates - by Bioz Stars, 2020-04
    90/100 stars
      Buy from Supplier

    92
    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
    https://www.bioz.com/result/middlebrook 7h10 plates/product/Becton Dickinson
    Average 92 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 plates - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    94
    Difco middlebrook 7h10
    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, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 178 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10/product/Difco
    Average 94 stars, based on 178 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    94
    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 140 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10/product/Becton Dickinson
    Average 94 stars, based on 140 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    93
    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 73 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 plates/product/MiddleBrook Pharmaceuticals
    Average 93 stars, based on 73 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 plates - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    87
    Millipore 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 Millipore, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 plates/product/Millipore
    Average 87 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 plates - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    91
    Molecular Toxicology Inc middlebrook 7h10 agar
    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 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
    https://www.bioz.com/result/middlebrook 7h10 agar/product/Molecular Toxicology Inc
    Average 91 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 agar - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    93
    Difco middlebrook 7h10 medium
    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 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
    https://www.bioz.com/result/middlebrook 7h10 medium/product/Difco
    Average 93 stars, based on 152 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 medium - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    93
    Becton Dickinson middlebrook 7h10 medium
    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 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
    https://www.bioz.com/result/middlebrook 7h10 medium/product/Becton Dickinson
    Average 93 stars, based on 29 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 medium - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    86
    Becton Dickinson middlebrook 7h10 7h11 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 7h11 Plates, 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
    https://www.bioz.com/result/middlebrook 7h10 7h11 plates/product/Becton Dickinson
    Average 86 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 7h11 plates - by Bioz Stars, 2020-04
    86/100 stars
      Buy from Supplier

    97
    MiddleBrook Pharmaceuticals middlebrook 7h10
    Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto <t>Middlebrook</t> <t>7H10</t> agar
    Middlebrook 7h10, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 97/100, based on 1221 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10/product/MiddleBrook Pharmaceuticals
    Average 97 stars, based on 1221 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    92
    MiddleBrook Pharmaceuticals middlebrook 7h10 oadc plates
    Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto <t>Middlebrook</t> <t>7H10</t> agar
    Middlebrook 7h10 Oadc Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 92/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 oadc plates/product/MiddleBrook Pharmaceuticals
    Average 92 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 oadc plates - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    92
    MiddleBrook Pharmaceuticals middlebrook 7h10 quad plates
    Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto <t>Middlebrook</t> <t>7H10</t> agar
    Middlebrook 7h10 Quad Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 92/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 quad plates/product/MiddleBrook Pharmaceuticals
    Average 92 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 quad plates - by Bioz Stars, 2020-04
    92/100 stars
      Buy from Supplier

    87
    MiddleBrook Pharmaceuticals middlebrook 7h10 oadc hyg plates
    Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto <t>Middlebrook</t> <t>7H10</t> agar
    Middlebrook 7h10 Oadc Hyg Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 87/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 oadc hyg plates/product/MiddleBrook Pharmaceuticals
    Average 87 stars, based on 2 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 oadc hyg plates - by Bioz Stars, 2020-04
    87/100 stars
      Buy from Supplier

    93
    MiddleBrook Pharmaceuticals middlebrook 7h10 oadc
    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
    https://www.bioz.com/result/middlebrook 7h10 oadc/product/MiddleBrook Pharmaceuticals
    Average 93 stars, based on 33 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 oadc - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    80
    MiddleBrook Pharmaceuticals fresh middlebrook 7h10 agar plate
    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
    Fresh Middlebrook 7h10 Agar Plate, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 80/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/fresh middlebrook 7h10 agar plate/product/MiddleBrook Pharmaceuticals
    Average 80 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    fresh middlebrook 7h10 agar plate - by Bioz Stars, 2020-04
    80/100 stars
      Buy from Supplier

    84
    MiddleBrook Pharmaceuticals prepared middlebrook 7h10 agar plates
    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
    Prepared Middlebrook 7h10 Agar Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/prepared middlebrook 7h10 agar plates/product/MiddleBrook Pharmaceuticals
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    prepared middlebrook 7h10 agar plates - by Bioz Stars, 2020-04
    84/100 stars
      Buy from Supplier

    84
    BVA Scientific middlebrook 7h10 agar
    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 Agar, supplied by BVA Scientific, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 agar/product/BVA Scientific
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 agar - by Bioz Stars, 2020-04
    84/100 stars
      Buy from Supplier

    91
    MiddleBrook Pharmaceuticals middlebrook 7h10 adn
    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 Adn, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 adn/product/MiddleBrook Pharmaceuticals
    Average 91 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 adn - by Bioz Stars, 2020-04
    91/100 stars
      Buy from Supplier

    94
    MiddleBrook Pharmaceuticals middlebrook 7h10 panta
    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 Panta, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 panta/product/MiddleBrook Pharmaceuticals
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 panta - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    93
    MiddleBrook Pharmaceuticals no drug 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
    No Drug Middlebrook 7h10, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 8 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/no drug middlebrook 7h10/product/MiddleBrook Pharmaceuticals
    Average 93 stars, based on 8 article reviews
    Price from $9.99 to $1999.99
    no drug middlebrook 7h10 - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    94
    Merck & Co middlebrook 7h10 agar
    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 Agar, supplied by Merck & Co, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 agar/product/Merck & Co
    Average 94 stars, based on 5 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 agar - by Bioz Stars, 2020-04
    94/100 stars
      Buy from Supplier

    93
    MiddleBrook Pharmaceuticals oadc supplemented middlebrook 7h10 agar plates
    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
    Oadc Supplemented Middlebrook 7h10 Agar Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/oadc supplemented middlebrook 7h10 agar plates/product/MiddleBrook Pharmaceuticals
    Average 93 stars, based on 6 article reviews
    Price from $9.99 to $1999.99
    oadc supplemented middlebrook 7h10 agar plates - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    93
    MiddleBrook Pharmaceuticals rif free middlebrook 7h10 agar plates
    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
    Rif Free Middlebrook 7h10 Agar Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/rif free middlebrook 7h10 agar plates/product/MiddleBrook Pharmaceuticals
    Average 93 stars, based on 10 article reviews
    Price from $9.99 to $1999.99
    rif free middlebrook 7h10 agar plates - by Bioz Stars, 2020-04
    93/100 stars
      Buy from Supplier

    84
    MiddleBrook Pharmaceuticals drug free middlebrook 7h10 agar plates
    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
    Drug Free Middlebrook 7h10 Agar Plates, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 84/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/drug free middlebrook 7h10 agar plates/product/MiddleBrook Pharmaceuticals
    Average 84 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    drug free middlebrook 7h10 agar plates - by Bioz Stars, 2020-04
    84/100 stars
      Buy from Supplier

    85
    MiddleBrook Pharmaceuticals 100 mm middlebrook 7h10 agar plate
    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
    100 Mm Middlebrook 7h10 Agar Plate, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 85/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/100 mm middlebrook 7h10 agar plate/product/MiddleBrook Pharmaceuticals
    Average 85 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
    100 mm middlebrook 7h10 agar plate - by Bioz Stars, 2020-04
    85/100 stars
      Buy from Supplier

    97
    MiddleBrook Pharmaceuticals middlebrook 7h10 medium
    Screening for LOS biosynthetic mutants in M. marinum . A , selection of rough morphotypes. Screening the Tn library on <t>Middlebrook</t> <t>7H10</t> led to six rough variants carrying Tn insertions in losA ( MMAR _ 2313 ) (two independent mutants), MMAR _ 2315 , MMAR _ 2319
    Middlebrook 7h10 Medium, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 97/100, based on 782 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/middlebrook 7h10 medium/product/MiddleBrook Pharmaceuticals
    Average 97 stars, based on 782 article reviews
    Price from $9.99 to $1999.99
    middlebrook 7h10 medium - by Bioz Stars, 2020-04
    97/100 stars
      Buy from Supplier

    Image Search Results


    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

    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

    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

    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

    Msmeg-PE11 are more resistant to SDS, lysozyme, hydrogen peroxide, low pH and antibiotics treatment when compared with Msmeg-pVV . The Msmeg-pVV or Msmeg-PE11 were either treated with medium alone or subjected to ( a ) 0.1% SDS, ( b ) lysozyme, ( c ) 5 mM hydrogen peroxide, ( d ) low pH (5.5 for 24 h), ( e ) 20 μg/ml ethambutol, ( f ) 20 μg/ml isoniazid, ( g ) rifampicin (20 μg/ml), ( h ) vancomycin (5 μg/ml) and ( i ) ampicillin (750 μg/ml) treatment. Bacterial CFUs were counted on 7H10 agar plates. Data are representative of mean ± SD of three different experiments.

    Journal: Scientific Reports

    Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence

    doi: 10.1038/srep21624

    Figure Lengend Snippet: Msmeg-PE11 are more resistant to SDS, lysozyme, hydrogen peroxide, low pH and antibiotics treatment when compared with Msmeg-pVV . The Msmeg-pVV or Msmeg-PE11 were either treated with medium alone or subjected to ( a ) 0.1% SDS, ( b ) lysozyme, ( c ) 5 mM hydrogen peroxide, ( d ) low pH (5.5 for 24 h), ( e ) 20 μg/ml ethambutol, ( f ) 20 μg/ml isoniazid, ( g ) rifampicin (20 μg/ml), ( h ) vancomycin (5 μg/ml) and ( i ) ampicillin (750 μg/ml) treatment. Bacterial CFUs were counted on 7H10 agar plates. Data are representative of mean ± SD of three different experiments.

    Article Snippet: The mouse tissue homogenates were diluted in PBS-T, and 100 μl aliquots from each dilution were plated on Middlebrook 7H10 agar plate containing 10% OADC, 25 μg/ml kanamycin and 50 μg/ml of hygromycin B.

    Techniques:

    PE11 alters colony morphology, surface architecture and width of M. smegmatis . ( a ) Msmeg-pVV or Msmeg-PE11 bacteria were plated on 7H10 agar plates supplemented with OADC and incubated for 5–6 days and the mycobacterial colonies were photographed. ( b ) For transmission electron microscopy (TEM) analysis, Msmeg-pVV or Msmeg-PE11 strains were cultured on 7H10 agar for 5–6 days and surface architecture of these bacteria were analyzed. Scale bar, 100 nm. ( c ) In another experiment, Msmeg-pVV or Msmeg-PE11 bacteria were harvested for scanning electron microscopy (SEM) analysis to measure the diameters of Msmeg-pVV or Msmeg-PE11 and mean width of 100 bacilli each of Msmeg-pVV and Msmeg-PE11 is graphically represented in nm.

    Journal: Scientific Reports

    Article Title: PE11, a PE/PPE family protein of Mycobacterium tuberculosis is involved in cell wall remodeling and virulence

    doi: 10.1038/srep21624

    Figure Lengend Snippet: PE11 alters colony morphology, surface architecture and width of M. smegmatis . ( a ) Msmeg-pVV or Msmeg-PE11 bacteria were plated on 7H10 agar plates supplemented with OADC and incubated for 5–6 days and the mycobacterial colonies were photographed. ( b ) For transmission electron microscopy (TEM) analysis, Msmeg-pVV or Msmeg-PE11 strains were cultured on 7H10 agar for 5–6 days and surface architecture of these bacteria were analyzed. Scale bar, 100 nm. ( c ) In another experiment, Msmeg-pVV or Msmeg-PE11 bacteria were harvested for scanning electron microscopy (SEM) analysis to measure the diameters of Msmeg-pVV or Msmeg-PE11 and mean width of 100 bacilli each of Msmeg-pVV and Msmeg-PE11 is graphically represented in nm.

    Article Snippet: The mouse tissue homogenates were diluted in PBS-T, and 100 μl aliquots from each dilution were plated on Middlebrook 7H10 agar plate containing 10% OADC, 25 μg/ml kanamycin and 50 μg/ml of hygromycin B.

    Techniques: Incubation, Transmission Assay, Electron Microscopy, Transmission Electron Microscopy, Cell Culture

    Survival of nonreplicating (NR) cultures of M. tuberculosis after exposure to drugs (alone and in combination) as estimated by CFU counts. Twelve-day-old (H12) cultures adjusted to pH 7.3 were incubated with drugs. Rifampin (R), 8 μg/ml; sutezolid (SZ), 1 μg/ml; nitazoxanide (NZ), 10 μg/ml; PA-824 (pretomanid; PA), 2 μg/ml; isoniazid (I), 2 μg/ml; pyrazinamide (Z), 100 μg/ml; ethambutol (E), 4 μg/ml. Dashed lines indicate the limit of detection (5 CFU/ml). Means and standard deviations from two experiments are shown. In each experiment, two Wayne tubes per condition per time point were used, from which at least two 7H10 agar plates were inoculated. (A) Single drugs, 2-drug combinations, and R-I-Z-E; (B) 3- and 4-drug combinations.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: The Combination Rifampin-Nitazoxanide, but Not Rifampin-Isoniazid-Pyrazinamide-Ethambutol, Kills Dormant Mycobacterium tuberculosis in Hypoxia at Neutral pH

    doi: 10.1128/AAC.00273-19

    Figure Lengend Snippet: Survival of nonreplicating (NR) cultures of M. tuberculosis after exposure to drugs (alone and in combination) as estimated by CFU counts. Twelve-day-old (H12) cultures adjusted to pH 7.3 were incubated with drugs. Rifampin (R), 8 μg/ml; sutezolid (SZ), 1 μg/ml; nitazoxanide (NZ), 10 μg/ml; PA-824 (pretomanid; PA), 2 μg/ml; isoniazid (I), 2 μg/ml; pyrazinamide (Z), 100 μg/ml; ethambutol (E), 4 μg/ml. Dashed lines indicate the limit of detection (5 CFU/ml). Means and standard deviations from two experiments are shown. In each experiment, two Wayne tubes per condition per time point were used, from which at least two 7H10 agar plates were inoculated. (A) Single drugs, 2-drug combinations, and R-I-Z-E; (B) 3- and 4-drug combinations.

    Article Snippet: Every week, 1 ml of NR culture was washed twice and resuspended in 1 ml of DTAB, and 0.2 ml was inoculated in Middlebrook 7H10 agar plates for CFU determination and in liquid medium (Bactec MGIT 960 system) for determination of the number of days to reach a growth unit of ≥75 (day to positivity [DTP]) ( ).

    Techniques: Incubation

    Integration of phoA into the chromosomes of M. smegmatis wild-type and porin mutants. (A) The phosphatase activity of M. smegmatis grown in liquid culture was determined by using p NPP. Black and gray bars indicate the phosphatase activity of whole and lysed cells, respectively. Error bars indicate standard deviations. (B) Phosphatase activity of M. smegmatis on Middlebrook 7H10 agar plates containing Tween 80 and BCIP. After 5 days of incubation, pictures of colonies were taken with a Zeiss stereomicroscope Stemi 2000-C using the same magnification for all strains. In both experiments, the strains were SMR5 (wild-type) and the porin mutants MN01 (Δ mspA ) and ML10 (Δ mspA/mspC ) and derivatives with an phoA expression cassette integrated at the attachment site of the phage L5 ( attB :: phoA ). The blue color indicates the cleavage of BCIP by PhoA.

    Journal: Journal of Bacteriology

    Article Title: Porins Are Required for Uptake of Phosphates by Mycobacterium smegmatis ▿

    doi: 10.1128/JB.01600-06

    Figure Lengend Snippet: Integration of phoA into the chromosomes of M. smegmatis wild-type and porin mutants. (A) The phosphatase activity of M. smegmatis grown in liquid culture was determined by using p NPP. Black and gray bars indicate the phosphatase activity of whole and lysed cells, respectively. Error bars indicate standard deviations. (B) Phosphatase activity of M. smegmatis on Middlebrook 7H10 agar plates containing Tween 80 and BCIP. After 5 days of incubation, pictures of colonies were taken with a Zeiss stereomicroscope Stemi 2000-C using the same magnification for all strains. In both experiments, the strains were SMR5 (wild-type) and the porin mutants MN01 (Δ mspA ) and ML10 (Δ mspA/mspC ) and derivatives with an phoA expression cassette integrated at the attachment site of the phage L5 ( attB :: phoA ). The blue color indicates the cleavage of BCIP by PhoA.

    Article Snippet: After selection on 7H10 plates containing kanamycin, 4 ml of Middlebrook 7H9 medium with kanamycin was inoculated and incubated for 12 h to 20 h at 37°C on a roller drum.

    Techniques: Activity Assay, Incubation, Expressing

    Porin-dependent growth of M. smegmatis on low-phosphate plates. (A to D) M. smegmatis SMR5 (wt) and ML10 (Δ mspA Δ mspC ) were streaked on self-made 7H10 agar plates containing different concentrations of phosphate and BCIP. The plates were incubated at 37°C for 6 days and were scanned (V700 Photo, Epson) each day. Scans of plates after 3 (A and C) and 6 days (B and D) of incubation are shown. The strains on the plates are as follows: streak 1, SMR5/pMS2 (control); streak 2, SMR5/pML440 ( phoA ); streak 3, ML10/pMS2; streak 4, ML10/pML440; and streak 5, ML10/pMN016 ( mspA ). (E to L) M. smegmatis SMR5 and ML10 were filtered through a 5-μm filter to obtain single cells and plated on self-made 7H10 agar plates containing different concentrations of phosphate and BCIP. Pictures of single colonies after 8 days of incubation at 37°C were taken at a 16-fold magnification using a Zeiss stereomicroscope Stemi 2000-C.

    Journal: Journal of Bacteriology

    Article Title: Porins Are Required for Uptake of Phosphates by Mycobacterium smegmatis ▿

    doi: 10.1128/JB.01600-06

    Figure Lengend Snippet: Porin-dependent growth of M. smegmatis on low-phosphate plates. (A to D) M. smegmatis SMR5 (wt) and ML10 (Δ mspA Δ mspC ) were streaked on self-made 7H10 agar plates containing different concentrations of phosphate and BCIP. The plates were incubated at 37°C for 6 days and were scanned (V700 Photo, Epson) each day. Scans of plates after 3 (A and C) and 6 days (B and D) of incubation are shown. The strains on the plates are as follows: streak 1, SMR5/pMS2 (control); streak 2, SMR5/pML440 ( phoA ); streak 3, ML10/pMS2; streak 4, ML10/pML440; and streak 5, ML10/pMN016 ( mspA ). (E to L) M. smegmatis SMR5 and ML10 were filtered through a 5-μm filter to obtain single cells and plated on self-made 7H10 agar plates containing different concentrations of phosphate and BCIP. Pictures of single colonies after 8 days of incubation at 37°C were taken at a 16-fold magnification using a Zeiss stereomicroscope Stemi 2000-C.

    Article Snippet: After selection on 7H10 plates containing kanamycin, 4 ml of Middlebrook 7H9 medium with kanamycin was inoculated and incubated for 12 h to 20 h at 37°C on a roller drum.

    Techniques: Incubation

    Porin-dependent phosphatase activity of M. smegmatis . (A) The phosphatase activity of M. smegmatis grown in liquid culture was determined using p NPP. Black and gray bars indicate the phosphatase activity in whole and lysed cells, respectively. The inset shows the endogenous phosphatase activity in the parent strains SMR5, MN01, and ML10. Error bars indicate standard deviations. (B) Phosphatase activity of M. smegmatis on Middlebrook 7H10 agar plates containing Tween 80 and BCIP. After 5 days of incubation, pictures of colonies were taken with a Zeiss stereomicroscope Stemi 2000-C using the same magnification for all strains. In both experiments, the strains were SMR5 (wild-type) and the porin mutants MN01 (Δ mspA ) and ML10 (Δ mspA Δ mspC ), each of which was transformed with the control plasmid pMS2 (control) and the phoA expression plasmid pML440 (pML440+ phoA ), respectively. The blue color indicates the cleavage of BCIP by PhoA.

    Journal: Journal of Bacteriology

    Article Title: Porins Are Required for Uptake of Phosphates by Mycobacterium smegmatis ▿

    doi: 10.1128/JB.01600-06

    Figure Lengend Snippet: Porin-dependent phosphatase activity of M. smegmatis . (A) The phosphatase activity of M. smegmatis grown in liquid culture was determined using p NPP. Black and gray bars indicate the phosphatase activity in whole and lysed cells, respectively. The inset shows the endogenous phosphatase activity in the parent strains SMR5, MN01, and ML10. Error bars indicate standard deviations. (B) Phosphatase activity of M. smegmatis on Middlebrook 7H10 agar plates containing Tween 80 and BCIP. After 5 days of incubation, pictures of colonies were taken with a Zeiss stereomicroscope Stemi 2000-C using the same magnification for all strains. In both experiments, the strains were SMR5 (wild-type) and the porin mutants MN01 (Δ mspA ) and ML10 (Δ mspA Δ mspC ), each of which was transformed with the control plasmid pMS2 (control) and the phoA expression plasmid pML440 (pML440+ phoA ), respectively. The blue color indicates the cleavage of BCIP by PhoA.

    Article Snippet: After selection on 7H10 plates containing kanamycin, 4 ml of Middlebrook 7H9 medium with kanamycin was inoculated and incubated for 12 h to 20 h at 37°C on a roller drum.

    Techniques: Activity Assay, Incubation, Transformation Assay, Plasmid Preparation, Expressing

    Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto Middlebrook 7H10 agar

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Novel Inhibitors of InhA Efficiently Kill Mycobacterium tuberculosis under Aerobic and Anaerobic Conditions ▿

    doi: 10.1128/AAC.00266-11

    Figure Lengend Snippet: Growth of M. tuberculosis H37Rv treated with CD13, CD39, CD59, and CD117 at 20 mg/liter. Cultures, grown in Sauton medium, were incubated at 37°C, and CFU were determined by plating 10-fold dilutions of each cultures onto Middlebrook 7H10 agar

    Article Snippet: M. tuberculosis H37Rv (109 cells) was plated on Middlebrook 7H10 plates supplemented with ADS and containing CD117 at 10 and 20 times the MIC.

    Techniques: Incubation

    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

    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

    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: The cultures were then diluted in Middlebrook 7H9 medium and plated on Middlebrook 7H10 ADN supplemented with and without 30 mM asparagine.

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

    Screening for LOS biosynthetic mutants in M. marinum . A , selection of rough morphotypes. Screening the Tn library on Middlebrook 7H10 led to six rough variants carrying Tn insertions in losA ( MMAR _ 2313 ) (two independent mutants), MMAR _ 2315 , MMAR _ 2319

    Journal: The Journal of Biological Chemistry

    Article Title: Increased Phagocytosis of Mycobacterium marinum Mutants Defective in Lipooligosaccharide Production

    doi: 10.1074/jbc.M113.525550

    Figure Lengend Snippet: Screening for LOS biosynthetic mutants in M. marinum . A , selection of rough morphotypes. Screening the Tn library on Middlebrook 7H10 led to six rough variants carrying Tn insertions in losA ( MMAR _ 2313 ) (two independent mutants), MMAR _ 2315 , MMAR _ 2319

    Article Snippet: Serial dilutions were plated onto Middlebrook 7H10 medium supplemented with 10% OADC.

    Techniques: Selection