7h10 agar Search Results


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  • 99
    Thermo Fisher 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 Thermo Fisher, used in various techniques. Bioz Stars score: 99/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Millipore 7h10 agar base
    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
    7h10 Agar Base, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 9 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Difco 7h10 agar
    Rel Mtb -mediated (p)ppGpp hydrolysis is required for growth in vitro and maintenance of ATP and GTP levels when (p)ppGpp is being produced. (A and B) An M. tuberculosis Δ rel Mtb strain containing an episomal vector expressing the WT TetR that induces expression of rel Mtb H80A from a cassette integrated at the attB site in the genome in the presence of ATc (Tet-Rel Mtb H80A strain, designated H80A here) and a WT M. tuberculosis strain containing an empty vector in the attB site and transformed with the same TetR-expressing plasmid (control strain). (A) Transcript levels in exponential-growth-phase cultures in liquid 7H9 media. rel Mtb H80A transcripts from the cassette at the attB site were detected in the Tet-Rel Mtb H80A strain, and transcripts from the endogenous rel Mtb gene were detected in the control strain. The first comparison shows the ratio of transcript levels in the presence compared to the absence of ATc in each strain (+ATc/−ATc), and it illustrates the induction of rel Mtb H80A in the Tet-Rel Mtb H80A strain when exposed to ATc and the unresponsiveness of the rel Mtb gene to ATc in the control strain. The second comparison shows the ratio of transcript levels in the Tet-Rel Mtb H80A strain compared to the control strain (H80A/control) under each condition to illustrate the low level of transcription of rel Mtb H80A in the absence of ATc compared to that of induced cultures. (B) M. tuberculosis Tet-Rel Mtb H80A and control strains were diluted and plated on <t>7H10</t> plates in the absence or presence of ATc. (C to G) M. smegmatis Δ rel Msm strains containing an episomal vector expressing the WT TetR that turns on expression of rel Mtb WT (Tet-Rel Mtb WT strain; designated the control) or rel Mtb H80A (Tet-Rel Mtb H80A strain; designated H80A here) from cassettes integrated at the attB site in the genome in the presence of ATc. (C and D) M. smegmatis strains were diluted and plated on 7H10 (C) and LB (D) plates in the absence or presence of ATc. (E) Graphic representation of the number of M. smegmatis CFU that grew in the presence compared to the absence of ATc when grown on LB, where all Tet-Rel Mtb H80A strain bacteria that grew in the presence of ATc on LB were suppressors and were no longer responsive to ATc treatment. Data are means ± SEM from 13 replicates. (F) ATP and GTP levels in M. smegmatis strains grown in the absence or presence of ATc for 6 h to an ODλ 600 of ∼0.6 in LB liquid media were measured by LC-MS/MS. The ratio of levels in the Tet-Rel Mtb H80A strain (H80A) to those in the Tet-Rel Mtb WT -expressing strain (control) under the same conditions is graphed. Data are the means ± SEM from 4 to 6 replicates. (E and F) The significance of differences was determined by calculating P values by Student's t tests; one asterisk indicates significance with a P value of
    7h10 Agar, supplied by Difco, used in various techniques. Bioz Stars score: 94/100, based on 277 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    96
    Becton Dickinson 7h10 agar
    CD40 engagement of DCs enhances control of Mtb infection. B6 DCs were left uninfected or infected with heat-killed Mtb in the presence or absence of CD40LT treatment (1 μg/ml), or infected with heat-killed hip1 mutant Mtb, each for 24 hours. Cells were washed twice and reconstituted in PBS to deliver 1x10 6 DC per mouse intratracheally. (A) Diagram of experimental design. Lung responses were assessed 6 and 12 days post-instillation. Mice were infected through the aerosol route with ~100 CFU Mtb 15 days post-instillation and bacterial burden was assessed 35 days (5 weeks) post-challenge. (B) Frequencies of CD44 + CD4 T cells in the lungs 6 and 12 days after intratracheal instillation of DCs. (C) Frequencies of IL-17 + (top) and IFN-γ + (bottom) CD4 + cells in the lungs 6 and 12 days after intratracheal instillation of DCs. Cells were stimulated with PMA (80 ng/ml) and ionomycin (500 ng/ml). (D) Lung bacterial burden 35 days post-challenge (overall day 50 post-DC intratracheal instillation). Bacterial burden was assessed by plating homogenized lungs on <t>7H10</t> agar plates and counting CFU. (E) Lung CD4 + IL-17 + and IFN-γ + frequencies at day 50 following Mtb whole cell lysate (10 μg/ml) restimulation. (F) Correlation plots showing association between lung bacterial burden and IL-17 (left) or IFN-γ (right) responses to WCL restimulation. A linear regression was utilized to generate a best-fit line and Spearman’s correlation coefficient calculated. 4–5 mice were used for each group. Statistical significance (B-E) was determined using one-way analysis of variance correcting for multiple comparisons. * p
    7h10 Agar, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 96/100, based on 136 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Becton Dickinson middlebrook agar 7h10
    CD40 engagement of DCs enhances control of Mtb infection. B6 DCs were left uninfected or infected with heat-killed Mtb in the presence or absence of CD40LT treatment (1 μg/ml), or infected with heat-killed hip1 mutant Mtb, each for 24 hours. Cells were washed twice and reconstituted in PBS to deliver 1x10 6 DC per mouse intratracheally. (A) Diagram of experimental design. Lung responses were assessed 6 and 12 days post-instillation. Mice were infected through the aerosol route with ~100 CFU Mtb 15 days post-instillation and bacterial burden was assessed 35 days (5 weeks) post-challenge. (B) Frequencies of CD44 + CD4 T cells in the lungs 6 and 12 days after intratracheal instillation of DCs. (C) Frequencies of IL-17 + (top) and IFN-γ + (bottom) CD4 + cells in the lungs 6 and 12 days after intratracheal instillation of DCs. Cells were stimulated with PMA (80 ng/ml) and ionomycin (500 ng/ml). (D) Lung bacterial burden 35 days post-challenge (overall day 50 post-DC intratracheal instillation). Bacterial burden was assessed by plating homogenized lungs on <t>7H10</t> agar plates and counting CFU. (E) Lung CD4 + IL-17 + and IFN-γ + frequencies at day 50 following Mtb whole cell lysate (10 μg/ml) restimulation. (F) Correlation plots showing association between lung bacterial burden and IL-17 (left) or IFN-γ (right) responses to WCL restimulation. A linear regression was utilized to generate a best-fit line and Spearman’s correlation coefficient calculated. 4–5 mice were used for each group. Statistical significance (B-E) was determined using one-way analysis of variance correcting for multiple comparisons. * p
    Middlebrook Agar 7h10, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 93/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    93
    Thermo Fisher 7h10 agar plates
    Kinyoun stain of patient isolate cultured on <t>7H10</t> agar at 35°C. Magnification, ×1,000.
    7h10 Agar Plates, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 60 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Millipore 7h10 agar
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar, supplied by Millipore, used in various techniques. Bioz Stars score: 92/100, based on 79 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    BVA Scientific 7h10 agar plates
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar Plates, supplied by BVA Scientific, used in various techniques. Bioz Stars score: 86/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Becton Dickinson middlesbrook 7h10 agar
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    Middlesbrook 7h10 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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    94
    Becton Dickinson 7h10 solid agar
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Solid Agar, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    86
    Difco 7h10 bacto agar
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Bacto Agar, supplied by Difco, used in various techniques. Bioz Stars score: 86/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Toxicology Inc 7h10 agar plates
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar Plates, supplied by Molecular Toxicology Inc, used in various techniques. Bioz Stars score: 86/100, based on 11 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    Becton Dickinson 7h10 broth agar
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Broth Agar, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 88/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Difco 7h10 agar m7h10
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar M7h10, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HiMedia Laboratories 7h10 agar plates
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar Plates, supplied by HiMedia Laboratories, 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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    Fisher Scientific 7h10 agar plates
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar Plates, supplied by Fisher Scientific, 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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    94
    Teknova 7h10 agar plates
    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on <t>7H10</t> agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.
    7h10 Agar Plates, supplied by Teknova, used in various techniques. Bioz Stars score: 94/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Difco solid 7h10 agar
    Isolation and visualization of viable mycobacteria in the RASC. (A) M . smegmatis :: gfp growth on solid <t>7H10</t> agar plates from the Six-Stage Viable Andersen Cascade Impactor after wet release of 200 μl diluted culture into the RASC (30 000, 3000 and 300 colony forming units—CFU). The columns indicate the particle sizes captured on each plate across the 6 stages of the impactor, and the rows indicate the estimated total number of CFU passing through the impactor. Each release was repeated three times and the mean and SD for each plate are presented below the typical growth pattern distribution seen in the particle release. In all the releases the sampling was run for 5 minutes at 28 l/min resulting in the potential total capture of 3000, 300 and 30 CFU respectively. (B) SEM (left) and fluorescent microscopy (right) of M . smegmatis :: gfp isolated on a PM10 impactor following experimental release.
    Solid 7h10 Agar, supplied by Difco, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Difco middlebook 7h10 agar
    Isolation and visualization of viable mycobacteria in the RASC. (A) M . smegmatis :: gfp growth on solid <t>7H10</t> agar plates from the Six-Stage Viable Andersen Cascade Impactor after wet release of 200 μl diluted culture into the RASC (30 000, 3000 and 300 colony forming units—CFU). The columns indicate the particle sizes captured on each plate across the 6 stages of the impactor, and the rows indicate the estimated total number of CFU passing through the impactor. Each release was repeated three times and the mean and SD for each plate are presented below the typical growth pattern distribution seen in the particle release. In all the releases the sampling was run for 5 minutes at 28 l/min resulting in the potential total capture of 3000, 300 and 30 CFU respectively. (B) SEM (left) and fluorescent microscopy (right) of M . smegmatis :: gfp isolated on a PM10 impactor following experimental release.
    Middlebook 7h10 Agar, supplied by Difco, 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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    94
    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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    96
    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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    93
    Difco 7h10 agar medium
    Increased release of membrane vesicles from the Δ pstA1 mutant requires RegX3. Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , Δ pstA1 Δ regX3 , and Δ pstA1 Δ regX3 pND regX3 strains were grown for 5 days in Sauton’s complete medium without Tween 80. (A) Cellular proteins (5 µg), secreted proteins depleted of MV (5 µg), and MV suspension (20 µl) were separated and analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analyses of culture supernatants. The results were normalized to numbers of CFU per milliliter determined from a control culture grown in Sauton’s medium with Tween 80 and plated on <t>7H10</t> medium. Data are means ± standard deviations for three independent cultures. **, P
    7h10 Agar Medium, supplied by Difco, used in various techniques. Bioz Stars score: 93/100, based on 117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Becton Dickinson 7h10 agar medium
    In vitro characterization of Δ rv2623 . (A) Growth curves of cultures inoculated (10 6 CFU/ml) into 7H9+10% OADC+0.05% Tween 80 (top) and in minimal Sauton's media (bottom); Erdman (closed boxes, solid line) and Δ rv2623 (open boxes, dashed line) cultures. Error bars represent the standard error of the means; each curve is a combination of at least three independent experiments. (B) Overexpression of Rv2623 in M. smegmatis . The top panel represents serial dilutions (1∶10) of the empty vector pMV261-containing negative control strain and Rv2623-overexpressing strain harboring pMV261:: rv2623 . Diluted stationary phase M. smegmatis culture was spotted (5 µl) onto solid <t>7H10</t> media supplemented with 10% OADC and kanamycin (40 µg/mL). Photographs were taken after three days incubation at 37°C. Growth of corresponding strains in liquid medium was assessed based on the time to detection determined using a BD BACTEC 9000MB system (bottom). The various strains were inoculated at 10 4 CFU/ml in triplicates. Data shown are representative of several independent experiments. ***p
    7h10 Agar Medium, supplied by Becton Dickinson, used in various techniques. Bioz Stars score: 94/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    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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    84
    Becton Dickinson cohn 7h10 agar 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).
    Cohn 7h10 Agar Plates, supplied by Becton Dickinson, 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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    Millipore 7h10 agar 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).
    7h10 Agar Plates, supplied by Millipore, 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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    Dot Scientific 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).
    7h10 Agar, supplied by Dot Scientific, used in various techniques. Bioz Stars score: 93/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals 7h10 agar
    Modified Middlebrook <t>7H10</t> 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
    7h10 Agar, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 93/100, based on 65 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Molecular Toxicology Inc middlebrook 7h10 agar
    Modified Middlebrook <t>7H10</t> 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
    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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    Clinical and Laboratory Standards Institute 7h10 agar
    Modified Middlebrook <t>7H10</t> 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
    7h10 Agar, supplied by Clinical and Laboratory Standards Institute, used in various techniques. Bioz Stars score: 86/100, based on 8 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

    Rel Mtb -mediated (p)ppGpp hydrolysis is required for growth in vitro and maintenance of ATP and GTP levels when (p)ppGpp is being produced. (A and B) An M. tuberculosis Δ rel Mtb strain containing an episomal vector expressing the WT TetR that induces expression of rel Mtb H80A from a cassette integrated at the attB site in the genome in the presence of ATc (Tet-Rel Mtb H80A strain, designated H80A here) and a WT M. tuberculosis strain containing an empty vector in the attB site and transformed with the same TetR-expressing plasmid (control strain). (A) Transcript levels in exponential-growth-phase cultures in liquid 7H9 media. rel Mtb H80A transcripts from the cassette at the attB site were detected in the Tet-Rel Mtb H80A strain, and transcripts from the endogenous rel Mtb gene were detected in the control strain. The first comparison shows the ratio of transcript levels in the presence compared to the absence of ATc in each strain (+ATc/−ATc), and it illustrates the induction of rel Mtb H80A in the Tet-Rel Mtb H80A strain when exposed to ATc and the unresponsiveness of the rel Mtb gene to ATc in the control strain. The second comparison shows the ratio of transcript levels in the Tet-Rel Mtb H80A strain compared to the control strain (H80A/control) under each condition to illustrate the low level of transcription of rel Mtb H80A in the absence of ATc compared to that of induced cultures. (B) M. tuberculosis Tet-Rel Mtb H80A and control strains were diluted and plated on 7H10 plates in the absence or presence of ATc. (C to G) M. smegmatis Δ rel Msm strains containing an episomal vector expressing the WT TetR that turns on expression of rel Mtb WT (Tet-Rel Mtb WT strain; designated the control) or rel Mtb H80A (Tet-Rel Mtb H80A strain; designated H80A here) from cassettes integrated at the attB site in the genome in the presence of ATc. (C and D) M. smegmatis strains were diluted and plated on 7H10 (C) and LB (D) plates in the absence or presence of ATc. (E) Graphic representation of the number of M. smegmatis CFU that grew in the presence compared to the absence of ATc when grown on LB, where all Tet-Rel Mtb H80A strain bacteria that grew in the presence of ATc on LB were suppressors and were no longer responsive to ATc treatment. Data are means ± SEM from 13 replicates. (F) ATP and GTP levels in M. smegmatis strains grown in the absence or presence of ATc for 6 h to an ODλ 600 of ∼0.6 in LB liquid media were measured by LC-MS/MS. The ratio of levels in the Tet-Rel Mtb H80A strain (H80A) to those in the Tet-Rel Mtb WT -expressing strain (control) under the same conditions is graphed. Data are the means ± SEM from 4 to 6 replicates. (E and F) The significance of differences was determined by calculating P values by Student's t tests; one asterisk indicates significance with a P value of

    Journal: Journal of Bacteriology

    Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp

    doi: 10.1128/JB.00759-13

    Figure Lengend Snippet: Rel Mtb -mediated (p)ppGpp hydrolysis is required for growth in vitro and maintenance of ATP and GTP levels when (p)ppGpp is being produced. (A and B) An M. tuberculosis Δ rel Mtb strain containing an episomal vector expressing the WT TetR that induces expression of rel Mtb H80A from a cassette integrated at the attB site in the genome in the presence of ATc (Tet-Rel Mtb H80A strain, designated H80A here) and a WT M. tuberculosis strain containing an empty vector in the attB site and transformed with the same TetR-expressing plasmid (control strain). (A) Transcript levels in exponential-growth-phase cultures in liquid 7H9 media. rel Mtb H80A transcripts from the cassette at the attB site were detected in the Tet-Rel Mtb H80A strain, and transcripts from the endogenous rel Mtb gene were detected in the control strain. The first comparison shows the ratio of transcript levels in the presence compared to the absence of ATc in each strain (+ATc/−ATc), and it illustrates the induction of rel Mtb H80A in the Tet-Rel Mtb H80A strain when exposed to ATc and the unresponsiveness of the rel Mtb gene to ATc in the control strain. The second comparison shows the ratio of transcript levels in the Tet-Rel Mtb H80A strain compared to the control strain (H80A/control) under each condition to illustrate the low level of transcription of rel Mtb H80A in the absence of ATc compared to that of induced cultures. (B) M. tuberculosis Tet-Rel Mtb H80A and control strains were diluted and plated on 7H10 plates in the absence or presence of ATc. (C to G) M. smegmatis Δ rel Msm strains containing an episomal vector expressing the WT TetR that turns on expression of rel Mtb WT (Tet-Rel Mtb WT strain; designated the control) or rel Mtb H80A (Tet-Rel Mtb H80A strain; designated H80A here) from cassettes integrated at the attB site in the genome in the presence of ATc. (C and D) M. smegmatis strains were diluted and plated on 7H10 (C) and LB (D) plates in the absence or presence of ATc. (E) Graphic representation of the number of M. smegmatis CFU that grew in the presence compared to the absence of ATc when grown on LB, where all Tet-Rel Mtb H80A strain bacteria that grew in the presence of ATc on LB were suppressors and were no longer responsive to ATc treatment. Data are means ± SEM from 13 replicates. (F) ATP and GTP levels in M. smegmatis strains grown in the absence or presence of ATc for 6 h to an ODλ 600 of ∼0.6 in LB liquid media were measured by LC-MS/MS. The ratio of levels in the Tet-Rel Mtb H80A strain (H80A) to those in the Tet-Rel Mtb WT -expressing strain (control) under the same conditions is graphed. Data are the means ± SEM from 4 to 6 replicates. (E and F) The significance of differences was determined by calculating P values by Student's t tests; one asterisk indicates significance with a P value of

    Article Snippet: Bacterial burden was determined by plating serial dilutions of lung and spleen homogenates onto 7H10 agar plates in the absence or presence of kanamycin and streptomycin.

    Techniques: In Vitro, Produced, Plasmid Preparation, Expressing, Transformation Assay, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

    Rv1366 is not necessary for M. tuberculosis growth or biofilm formation in vitro . (A) Southern blot analysis of WT M. tuberculosis Erdman (lane 1) and the Δ rv1366 strain (lanes 2 to 4) using EcoRI-digested genomic DNA and a probe that spans nucleotides 1537706 to 1538406 of the M. tuberculosis genome. EcoRI digestion of WT M. tuberculosis yields a band at 5,083 bp. EcoRI digestion of Δ rv1366 results in a 2,226-bp band due to the replacement of the rv1366 gene with a hygromycin resistance cassette. (B) Southern blot analysis of WT M. tuberculosis Erdman (lane 1) and the Δ rel Mtb Δ rv1366 strain (lanes 2 to 6) using SmaI-digested genomic DNA and a probe that spans nucleotides 2910191 to 2910844 of the M. tuberculosis genome. SmaI digestion of WT M. tuberculosis yields a band at 2,257 bp. SmaI digestion of the Δ rel Mtb Δ rv1366 strain results in a 4,769-bp band due to the replacement of the rel Mtb gene with a hygromycin resistance cassette. (C and D) M. tuberculosis Δ rel Mtb , Δ rv1366 , and Δ rel Mtb Δ rv1366 strains containing a vector that constitutively expresses Rel Mtb WT or Rv1366 or containing an empty vector (−) integrated at the attB site in the genome were normalized to the same ODλ 600 and either diluted and plated on 7H10 plates (C) or grown in Sauton's media in static, biofilm-forming conditions in a 24-well dish (D). (C) 7H10 plates were incubated for 3 weeks at 37°C. (D) Biofilms were incubated for 4 weeks at 37°C. The parent strain used is labeled above each photo, and the protein expressed from the attB site is labeled below the photo.

    Journal: Journal of Bacteriology

    Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp

    doi: 10.1128/JB.00759-13

    Figure Lengend Snippet: Rv1366 is not necessary for M. tuberculosis growth or biofilm formation in vitro . (A) Southern blot analysis of WT M. tuberculosis Erdman (lane 1) and the Δ rv1366 strain (lanes 2 to 4) using EcoRI-digested genomic DNA and a probe that spans nucleotides 1537706 to 1538406 of the M. tuberculosis genome. EcoRI digestion of WT M. tuberculosis yields a band at 5,083 bp. EcoRI digestion of Δ rv1366 results in a 2,226-bp band due to the replacement of the rv1366 gene with a hygromycin resistance cassette. (B) Southern blot analysis of WT M. tuberculosis Erdman (lane 1) and the Δ rel Mtb Δ rv1366 strain (lanes 2 to 6) using SmaI-digested genomic DNA and a probe that spans nucleotides 2910191 to 2910844 of the M. tuberculosis genome. SmaI digestion of WT M. tuberculosis yields a band at 2,257 bp. SmaI digestion of the Δ rel Mtb Δ rv1366 strain results in a 4,769-bp band due to the replacement of the rel Mtb gene with a hygromycin resistance cassette. (C and D) M. tuberculosis Δ rel Mtb , Δ rv1366 , and Δ rel Mtb Δ rv1366 strains containing a vector that constitutively expresses Rel Mtb WT or Rv1366 or containing an empty vector (−) integrated at the attB site in the genome were normalized to the same ODλ 600 and either diluted and plated on 7H10 plates (C) or grown in Sauton's media in static, biofilm-forming conditions in a 24-well dish (D). (C) 7H10 plates were incubated for 3 weeks at 37°C. (D) Biofilms were incubated for 4 weeks at 37°C. The parent strain used is labeled above each photo, and the protein expressed from the attB site is labeled below the photo.

    Article Snippet: Bacterial burden was determined by plating serial dilutions of lung and spleen homogenates onto 7H10 agar plates in the absence or presence of kanamycin and streptomycin.

    Techniques: In Vitro, Southern Blot, Plasmid Preparation, Incubation, Labeling

    Rel Mtb -mediated (p)ppGpp synthesis is necessary for optimal M. tuberculosis growth and biofilm formation in vitro. M. tuberculosis Δ rel Mtb strains containing a vector that constitutively expresses Rel Mtb WT (WT) or Rel Mtb H344Y (H344Y) or containing an empty vector (−) integrated at the attB site in the genome were normalized to the same ODλ 600 and either diluted and plated on 7H10 plates (A) or grown in Sauton's medium in static, biofilm-forming conditions in a 24-well dish (B). (A) 7H10 plates were incubated for 3 weeks at 37°C. (B) Biofilms were incubated for 4 weeks at 37°C. The parent strain used is labeled above each photo, and the protein expressed from the attB site is labeled below the photo.

    Journal: Journal of Bacteriology

    Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp

    doi: 10.1128/JB.00759-13

    Figure Lengend Snippet: Rel Mtb -mediated (p)ppGpp synthesis is necessary for optimal M. tuberculosis growth and biofilm formation in vitro. M. tuberculosis Δ rel Mtb strains containing a vector that constitutively expresses Rel Mtb WT (WT) or Rel Mtb H344Y (H344Y) or containing an empty vector (−) integrated at the attB site in the genome were normalized to the same ODλ 600 and either diluted and plated on 7H10 plates (A) or grown in Sauton's medium in static, biofilm-forming conditions in a 24-well dish (B). (A) 7H10 plates were incubated for 3 weeks at 37°C. (B) Biofilms were incubated for 4 weeks at 37°C. The parent strain used is labeled above each photo, and the protein expressed from the attB site is labeled below the photo.

    Article Snippet: Bacterial burden was determined by plating serial dilutions of lung and spleen homogenates onto 7H10 agar plates in the absence or presence of kanamycin and streptomycin.

    Techniques: In Vitro, Plasmid Preparation, Incubation, Labeling

    Rel Mtb (p)ppGpp hydrolase activity is essential for acute and chronic M. tuberculosis infection of mice. Bacterial titers in the lungs (A, E, and G) and spleens (B and H) of C57BL/6 mice infected with either control or Tet-Rel Mtb H80A (designated H80A in the figure) M. tuberculosis strains. The parent strain for the control is WT M. tuberculosis Erdman, and the parent strain for Tet-Rel Mtb H80A is M. tuberculosis Δ rel Mtb . Both strains express TetR from a plasmid that confers streptomycin resistance and contain a kanamycin resistance cassette integrated into the attB site that, in the case of Tet-Rel Mtb H80A , carries the rel Mtb H80A allele. The strain symbols in panel A are the same for all panels. (A to C) Mice were given normal mouse chow throughout infection. (E and F) Mice were administered doxycycline-containing mouse chow starting at day 1 postinfection (designated by the arrow). (G to I) Mice were administered doxycycline-containing mouse chow starting at day 63 postinfection (designated by the arrow). (C, F, and I) The ratio of CFU from the lungs or spleens grown on 7H10 plates containing streptomycin and kanamycin compared to 7H10 containing no antibiotics (No AB). ND denotes when no colonies were recovered after plating 5% of the lung homogenate (limit of detection, 20 CFU). Data are means ± SEM of 6 Tet-Rel Mtb H80A strain-infected mice and 3 control strain-infected mice per time point from two replicate experiments. (E, G, and H) The significance of differences were determined by calculating P values by Student's t tests; two asterisks indicate significance with a P value of

    Journal: Journal of Bacteriology

    Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp

    doi: 10.1128/JB.00759-13

    Figure Lengend Snippet: Rel Mtb (p)ppGpp hydrolase activity is essential for acute and chronic M. tuberculosis infection of mice. Bacterial titers in the lungs (A, E, and G) and spleens (B and H) of C57BL/6 mice infected with either control or Tet-Rel Mtb H80A (designated H80A in the figure) M. tuberculosis strains. The parent strain for the control is WT M. tuberculosis Erdman, and the parent strain for Tet-Rel Mtb H80A is M. tuberculosis Δ rel Mtb . Both strains express TetR from a plasmid that confers streptomycin resistance and contain a kanamycin resistance cassette integrated into the attB site that, in the case of Tet-Rel Mtb H80A , carries the rel Mtb H80A allele. The strain symbols in panel A are the same for all panels. (A to C) Mice were given normal mouse chow throughout infection. (E and F) Mice were administered doxycycline-containing mouse chow starting at day 1 postinfection (designated by the arrow). (G to I) Mice were administered doxycycline-containing mouse chow starting at day 63 postinfection (designated by the arrow). (C, F, and I) The ratio of CFU from the lungs or spleens grown on 7H10 plates containing streptomycin and kanamycin compared to 7H10 containing no antibiotics (No AB). ND denotes when no colonies were recovered after plating 5% of the lung homogenate (limit of detection, 20 CFU). Data are means ± SEM of 6 Tet-Rel Mtb H80A strain-infected mice and 3 control strain-infected mice per time point from two replicate experiments. (E, G, and H) The significance of differences were determined by calculating P values by Student's t tests; two asterisks indicate significance with a P value of

    Article Snippet: Bacterial burden was determined by plating serial dilutions of lung and spleen homogenates onto 7H10 agar plates in the absence or presence of kanamycin and streptomycin.

    Techniques: Activity Assay, Infection, Mouse Assay, Plasmid Preparation

    Course of M. tuberculosis aerosol challenge in the lungs of memory and naive mice. Memory immune (●) and naive (□) mice were infected with ∼60 M. tuberculosis bacilli via aerosol. The numbers of viable bacilli were determined by plating serial dilutions of lung homogenates onto 7H10 plates and counting the colonies after 3 weeks at 37°C. Each time point represents four mice, and the experiment was repeated once. The standard error bars are too small to see on this graph. ∗, P ≤ 0.001, comparing naive and memory mice at each time point.

    Journal: Infection and Immunity

    Article Title: CD8+ T Cells Participate in the Memory Immune Response to Mycobacterium tuberculosis

    doi: 10.1128/IAI.69.7.4320-4328.2001

    Figure Lengend Snippet: Course of M. tuberculosis aerosol challenge in the lungs of memory and naive mice. Memory immune (●) and naive (□) mice were infected with ∼60 M. tuberculosis bacilli via aerosol. The numbers of viable bacilli were determined by plating serial dilutions of lung homogenates onto 7H10 plates and counting the colonies after 3 weeks at 37°C. Each time point represents four mice, and the experiment was repeated once. The standard error bars are too small to see on this graph. ∗, P ≤ 0.001, comparing naive and memory mice at each time point.

    Article Snippet: Mice were infected intravenously (i.v.) via tail vein with 2 × 105 live bacilli in 100 μl or by aerosol with approximately 100 live bacilli as determined by viable counts on 7H10 agar plates (Difco Laboratories, Detroit, Mich.).

    Techniques: Mouse Assay, Infection

    a) M. smegmatis , M. abscessus , M. fortuitum subsp. fortuitum , M. chelonae , and M. goodii were patched onto 7H10 agar media (Difco) at pH 7.0, 6.0, 5.5 or 5.0 to determine pigment production . b) M. avium intracellulare and M. avium subsps. avium were patched onto 7H10 agar media at pH 7.0, 6.0, and 5.5 to determine pigment production. c) From left to right: acetone only, M. smegmatis pH 7.0 extracted w/acetone, M. smegmatis pH 6.0 extracted w/acetone, M. smegmatis pH 5.5 extracted with acetone. d) Fluorescence specific activity units of M. smegmatis bearing empty reporter plasmid pFPV27 or the M. smegmatis probable carotenoid promoter upstream of gfp ( pCar ) at pH 7.0, 6.0 or 5.5.

    Journal: BMC Research Notes

    Article Title: Acidochromogenicity is a common characteristic in nontuberculous mycobacteria

    doi: 10.1186/1756-0500-4-466

    Figure Lengend Snippet: a) M. smegmatis , M. abscessus , M. fortuitum subsp. fortuitum , M. chelonae , and M. goodii were patched onto 7H10 agar media (Difco) at pH 7.0, 6.0, 5.5 or 5.0 to determine pigment production . b) M. avium intracellulare and M. avium subsps. avium were patched onto 7H10 agar media at pH 7.0, 6.0, and 5.5 to determine pigment production. c) From left to right: acetone only, M. smegmatis pH 7.0 extracted w/acetone, M. smegmatis pH 6.0 extracted w/acetone, M. smegmatis pH 5.5 extracted with acetone. d) Fluorescence specific activity units of M. smegmatis bearing empty reporter plasmid pFPV27 or the M. smegmatis probable carotenoid promoter upstream of gfp ( pCar ) at pH 7.0, 6.0 or 5.5.

    Article Snippet: Isolated colonies of mycobacteria were patched onto 7H10 agar media (Difco) adjusted to pH 5.5 or 6.0 with HCl, or pH 7.0 with NaOH and containing 10% ADC (bovine serum albumin, dextrose and NaCl) and 0.2% glycerol and were incubated for 72 hours at 37°C in the dark.

    Techniques: Fluorescence, Activity Assay, Plasmid Preparation

    CD40 engagement of DCs enhances control of Mtb infection. B6 DCs were left uninfected or infected with heat-killed Mtb in the presence or absence of CD40LT treatment (1 μg/ml), or infected with heat-killed hip1 mutant Mtb, each for 24 hours. Cells were washed twice and reconstituted in PBS to deliver 1x10 6 DC per mouse intratracheally. (A) Diagram of experimental design. Lung responses were assessed 6 and 12 days post-instillation. Mice were infected through the aerosol route with ~100 CFU Mtb 15 days post-instillation and bacterial burden was assessed 35 days (5 weeks) post-challenge. (B) Frequencies of CD44 + CD4 T cells in the lungs 6 and 12 days after intratracheal instillation of DCs. (C) Frequencies of IL-17 + (top) and IFN-γ + (bottom) CD4 + cells in the lungs 6 and 12 days after intratracheal instillation of DCs. Cells were stimulated with PMA (80 ng/ml) and ionomycin (500 ng/ml). (D) Lung bacterial burden 35 days post-challenge (overall day 50 post-DC intratracheal instillation). Bacterial burden was assessed by plating homogenized lungs on 7H10 agar plates and counting CFU. (E) Lung CD4 + IL-17 + and IFN-γ + frequencies at day 50 following Mtb whole cell lysate (10 μg/ml) restimulation. (F) Correlation plots showing association between lung bacterial burden and IL-17 (left) or IFN-γ (right) responses to WCL restimulation. A linear regression was utilized to generate a best-fit line and Spearman’s correlation coefficient calculated. 4–5 mice were used for each group. Statistical significance (B-E) was determined using one-way analysis of variance correcting for multiple comparisons. * p

    Journal: PLoS Pathogens

    Article Title: Engaging the CD40-CD40L pathway augments T-helper cell responses and improves control of Mycobacterium tuberculosis infection

    doi: 10.1371/journal.ppat.1006530

    Figure Lengend Snippet: CD40 engagement of DCs enhances control of Mtb infection. B6 DCs were left uninfected or infected with heat-killed Mtb in the presence or absence of CD40LT treatment (1 μg/ml), or infected with heat-killed hip1 mutant Mtb, each for 24 hours. Cells were washed twice and reconstituted in PBS to deliver 1x10 6 DC per mouse intratracheally. (A) Diagram of experimental design. Lung responses were assessed 6 and 12 days post-instillation. Mice were infected through the aerosol route with ~100 CFU Mtb 15 days post-instillation and bacterial burden was assessed 35 days (5 weeks) post-challenge. (B) Frequencies of CD44 + CD4 T cells in the lungs 6 and 12 days after intratracheal instillation of DCs. (C) Frequencies of IL-17 + (top) and IFN-γ + (bottom) CD4 + cells in the lungs 6 and 12 days after intratracheal instillation of DCs. Cells were stimulated with PMA (80 ng/ml) and ionomycin (500 ng/ml). (D) Lung bacterial burden 35 days post-challenge (overall day 50 post-DC intratracheal instillation). Bacterial burden was assessed by plating homogenized lungs on 7H10 agar plates and counting CFU. (E) Lung CD4 + IL-17 + and IFN-γ + frequencies at day 50 following Mtb whole cell lysate (10 μg/ml) restimulation. (F) Correlation plots showing association between lung bacterial burden and IL-17 (left) or IFN-γ (right) responses to WCL restimulation. A linear regression was utilized to generate a best-fit line and Spearman’s correlation coefficient calculated. 4–5 mice were used for each group. Statistical significance (B-E) was determined using one-way analysis of variance correcting for multiple comparisons. * p

    Article Snippet: Briefly, bacteria were grown at 37°C in Middlebrook 7H9 broth or 7H10 agar supplemented with 10% oleic acid-albumin-dextrose-catalase (OADC) (Becton Dickinson, Franklin Lakes, NJ), 0.5% glycerol, and 0.05% Tween 80 (for broth), with the addition of 25 μg/ml kanamycin (Sigma-Aldrich, St. Louis, MO) for hip1 mutant Mtb.

    Techniques: Infection, Mutagenesis, Mouse Assay

    The anti-bacillus effect of vitamin B1 (VB1) in macrophages. (A,B) Bone marrow-derived macrophages were pretreated with phosphate buffer saline, VB1, Rosi, or mixture of VB1 and Rosi for 24 h and then challenged with Mycobacterium tuberculosis H37Rv (MOI 5) for 1 h. (A) Intracellular viable bacteria were detected with colony-forming unit (CFU) assays at 0, 1, 2, 3 day post-infection. (B) Intracellular viable bacteria were detected with CFU assays at 72 h post-infection. (C,D) PPAR-γ fl/fl or PPAR-γ fl/fl -Lys2-cre mice were infected with H37Rv (~200 bacteria/mouse). Oral administration with water (Ctrl), VB1, INH, or Rosi ( n = 5 mice/group) was started from the day after infection (day 1) and continued for 2 weeks. CFUs were obtained from the lung cell lysates by serial dilution and plating on 7H10 agars in triplicate. The colonies were counted after 4 weeks. (C) C57BL/6J mice. (D) PPAR-γ fl/fl or PPAR-γ fl/fl -Lys2-cre mice. Data shown are the mean ± SD. * P

    Journal: Frontiers in Immunology

    Article Title: Vitamin B1 Helps to Limit Mycobacterium tuberculosis Growth via Regulating Innate Immunity in a Peroxisome Proliferator-Activated Receptor-γ-Dependent Manner

    doi: 10.3389/fimmu.2018.01778

    Figure Lengend Snippet: The anti-bacillus effect of vitamin B1 (VB1) in macrophages. (A,B) Bone marrow-derived macrophages were pretreated with phosphate buffer saline, VB1, Rosi, or mixture of VB1 and Rosi for 24 h and then challenged with Mycobacterium tuberculosis H37Rv (MOI 5) for 1 h. (A) Intracellular viable bacteria were detected with colony-forming unit (CFU) assays at 0, 1, 2, 3 day post-infection. (B) Intracellular viable bacteria were detected with CFU assays at 72 h post-infection. (C,D) PPAR-γ fl/fl or PPAR-γ fl/fl -Lys2-cre mice were infected with H37Rv (~200 bacteria/mouse). Oral administration with water (Ctrl), VB1, INH, or Rosi ( n = 5 mice/group) was started from the day after infection (day 1) and continued for 2 weeks. CFUs were obtained from the lung cell lysates by serial dilution and plating on 7H10 agars in triplicate. The colonies were counted after 4 weeks. (C) C57BL/6J mice. (D) PPAR-γ fl/fl or PPAR-γ fl/fl -Lys2-cre mice. Data shown are the mean ± SD. * P

    Article Snippet: Bacterial burden was determined by plating serial dilutions of spleen and lung homogenates onto 7H10 agar plates (BD Biosciences, USA) with 10% OADC.

    Techniques: Derivative Assay, Infection, Mouse Assay, Serial Dilution

    The anti-bacillus effect of vitamin B1 (VB1) in mice with Mycobacterium tuberculosis infection. C57BL/6J mice were infected with H37Rv (~200 bacteria/mouse). Oral administration with water (Ctrl), VB1, and INH ( n = 15 mice/group) was started from the day after infection (day 1) and continued for 1, 2, and 4 weeks alternatively. The lungs and spleens were analyzed at indicated time. Colony-forming units (CFUs) were obtained from the lung and spleen cell lysates by serial dilution and plating on 7H10 agars in triplicate. The colonies were counted after 4 weeks. Data shown are the mean ± SD. * P

    Journal: Frontiers in Immunology

    Article Title: Vitamin B1 Helps to Limit Mycobacterium tuberculosis Growth via Regulating Innate Immunity in a Peroxisome Proliferator-Activated Receptor-γ-Dependent Manner

    doi: 10.3389/fimmu.2018.01778

    Figure Lengend Snippet: The anti-bacillus effect of vitamin B1 (VB1) in mice with Mycobacterium tuberculosis infection. C57BL/6J mice were infected with H37Rv (~200 bacteria/mouse). Oral administration with water (Ctrl), VB1, and INH ( n = 15 mice/group) was started from the day after infection (day 1) and continued for 1, 2, and 4 weeks alternatively. The lungs and spleens were analyzed at indicated time. Colony-forming units (CFUs) were obtained from the lung and spleen cell lysates by serial dilution and plating on 7H10 agars in triplicate. The colonies were counted after 4 weeks. Data shown are the mean ± SD. * P

    Article Snippet: Bacterial burden was determined by plating serial dilutions of spleen and lung homogenates onto 7H10 agar plates (BD Biosciences, USA) with 10% OADC.

    Techniques: Mouse Assay, Infection, Serial Dilution

    Kinyoun stain of patient isolate cultured on 7H10 agar at 35°C. Magnification, ×1,000.

    Journal: Journal of Clinical Microbiology

    Article Title: Brown-Pigmented Mycobacterium mageritense as a Cause of Prosthetic Valve Endocarditis and Bloodstream Infection

    doi: 10.1128/JCM.01041-15

    Figure Lengend Snippet: Kinyoun stain of patient isolate cultured on 7H10 agar at 35°C. Magnification, ×1,000.

    Article Snippet: Positive blood cultures were subcultured to Middlebrook 7H10 agar plates (Remel, Lenexa, KS) and Lowenstein-Jensen (LJ) agar slants (Becton Dickinson, Sparks, MD) and incubated at 35°C.

    Techniques: Staining, Cell Culture

    Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on 7H10 agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Aminoglycoside Cross-Resistance in Mycobacterium tuberculosis Due to Mutations in the 5? Untranslated Region of whiB7

    doi: 10.1128/AAC.02191-12

    Figure Lengend Snippet: Antibiotic resistance patterns of CDC1551 deletion strains. M. tuberculosis strains were streaked on 7H10 agar containing either 5 μg/ml KAN (A) or 2 μg/ml STR (B) and incubated for 28 days at 37°C.

    Article Snippet: The susceptibilities to KAN and STR were determined according to guidelines and definitions stated by the Clinical and Laboratory Standards Institute , using 7H10 agar containing KAN (Sigma) at concentrations of 1, 2, 3, 4, 5, 10, 15, 20, 25, 30, 35, 40, 50, and 80 μg/ml and STR, isoniazid (INH), capreomycin (CAP), rifampin (RIF), and ofloxacin (OFX) at 0.25, 0.5, 1, 2, 4, 8, 10, and 16 μg/ml.

    Techniques: Incubation

    Isolation and visualization of viable mycobacteria in the RASC. (A) M . smegmatis :: gfp growth on solid 7H10 agar plates from the Six-Stage Viable Andersen Cascade Impactor after wet release of 200 μl diluted culture into the RASC (30 000, 3000 and 300 colony forming units—CFU). The columns indicate the particle sizes captured on each plate across the 6 stages of the impactor, and the rows indicate the estimated total number of CFU passing through the impactor. Each release was repeated three times and the mean and SD for each plate are presented below the typical growth pattern distribution seen in the particle release. In all the releases the sampling was run for 5 minutes at 28 l/min resulting in the potential total capture of 3000, 300 and 30 CFU respectively. (B) SEM (left) and fluorescent microscopy (right) of M . smegmatis :: gfp isolated on a PM10 impactor following experimental release.

    Journal: PLoS ONE

    Article Title: Real-Time Investigation of Tuberculosis Transmission: Developing the Respiratory Aerosol Sampling Chamber (RASC)

    doi: 10.1371/journal.pone.0146658

    Figure Lengend Snippet: Isolation and visualization of viable mycobacteria in the RASC. (A) M . smegmatis :: gfp growth on solid 7H10 agar plates from the Six-Stage Viable Andersen Cascade Impactor after wet release of 200 μl diluted culture into the RASC (30 000, 3000 and 300 colony forming units—CFU). The columns indicate the particle sizes captured on each plate across the 6 stages of the impactor, and the rows indicate the estimated total number of CFU passing through the impactor. Each release was repeated three times and the mean and SD for each plate are presented below the typical growth pattern distribution seen in the particle release. In all the releases the sampling was run for 5 minutes at 28 l/min resulting in the potential total capture of 3000, 300 and 30 CFU respectively. (B) SEM (left) and fluorescent microscopy (right) of M . smegmatis :: gfp isolated on a PM10 impactor following experimental release.

    Article Snippet: For the pilot TB studies, the Andersen impactor contained solid 7H10 agar (Difco) plates supplemented with 0.2% glycerol and oleic acid/albumin/dextrose/catalase (OADC) complex.

    Techniques: Isolation, Sampling, Microscopy

    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:

    Increased release of membrane vesicles from the Δ pstA1 mutant requires RegX3. Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , Δ pstA1 Δ regX3 , and Δ pstA1 Δ regX3 pND regX3 strains were grown for 5 days in Sauton’s complete medium without Tween 80. (A) Cellular proteins (5 µg), secreted proteins depleted of MV (5 µg), and MV suspension (20 µl) were separated and analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analyses of culture supernatants. The results were normalized to numbers of CFU per milliliter determined from a control culture grown in Sauton’s medium with Tween 80 and plated on 7H10 medium. Data are means ± standard deviations for three independent cultures. **, P

    Journal: mBio

    Article Title: Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity

    doi: 10.1128/mBio.00778-18

    Figure Lengend Snippet: Increased release of membrane vesicles from the Δ pstA1 mutant requires RegX3. Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , Δ pstA1 Δ regX3 , and Δ pstA1 Δ regX3 pND regX3 strains were grown for 5 days in Sauton’s complete medium without Tween 80. (A) Cellular proteins (5 µg), secreted proteins depleted of MV (5 µg), and MV suspension (20 µl) were separated and analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analyses of culture supernatants. The results were normalized to numbers of CFU per milliliter determined from a control culture grown in Sauton’s medium with Tween 80 and plated on 7H10 medium. Data are means ± standard deviations for three independent cultures. **, P

    Article Snippet: Bacteria were routinely cultured in Middlebrook 7H9 liquid medium (Difco) supplemented with 10% albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD Biosciences) and 0.5% glycerol.

    Techniques: Mutagenesis, Western Blot

    Depletion of EccD 5 prevents ESX-5 secretion but does not affect membrane vesicle production. The eccD 5 Tet-OFF and Δ pstA1 eccD 5 Tet-OFF mutants were grown for 5 days in Sauton’s complete medium without Tween 80. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added as indicated to repress eccD 5 . (A) Cellular proteins (5 µg), secreted proteins depleted of MV (5 µg), and MV suspension (20 µl) were analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analysis of culture supernatants. Numbers of particles per milliliter were normalized to numbers of CFU per milliliter determined from a control culture of each strain grown with Tween 80 and plated on 7H10 complete medium. The data are means ± standard deviations for three independent cultures.

    Journal: mBio

    Article Title: Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity

    doi: 10.1128/mBio.00778-18

    Figure Lengend Snippet: Depletion of EccD 5 prevents ESX-5 secretion but does not affect membrane vesicle production. The eccD 5 Tet-OFF and Δ pstA1 eccD 5 Tet-OFF mutants were grown for 5 days in Sauton’s complete medium without Tween 80. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added as indicated to repress eccD 5 . (A) Cellular proteins (5 µg), secreted proteins depleted of MV (5 µg), and MV suspension (20 µl) were analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analysis of culture supernatants. Numbers of particles per milliliter were normalized to numbers of CFU per milliliter determined from a control culture of each strain grown with Tween 80 and plated on 7H10 complete medium. The data are means ± standard deviations for three independent cultures.

    Article Snippet: Bacteria were routinely cultured in Middlebrook 7H9 liquid medium (Difco) supplemented with 10% albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD Biosciences) and 0.5% glycerol.

    Techniques: Western Blot

    Increased vesicle release from the Δ pstA1 mutant is independent of VirR. Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , Δ virR , Δ pstA1 Δ virR , WT p virR , and Δ pstA1 p virR strains were grown for 5 days in Sauton’s complete medium without Tween 80. (A) Cellular proteins (5 µg) and MV suspension (20 µl) were analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analysis of culture supernatants. Numbers of particles per milliliter were normalized to numbers of CFU per milliliter determined from a control culture grown in Sauton’s medium with Tween 80 and plated on 7H10 medium. Data are means ± standard deviations for three independent cultures. The results of all statistical analyses are compared to WT levels unless otherwise indicated. *, P

    Journal: mBio

    Article Title: Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity

    doi: 10.1128/mBio.00778-18

    Figure Lengend Snippet: Increased vesicle release from the Δ pstA1 mutant is independent of VirR. Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , Δ virR , Δ pstA1 Δ virR , WT p virR , and Δ pstA1 p virR strains were grown for 5 days in Sauton’s complete medium without Tween 80. (A) Cellular proteins (5 µg) and MV suspension (20 µl) were analyzed by Western blotting to detect the indicated proteins. (B) Nanoparticle tracking analysis of culture supernatants. Numbers of particles per milliliter were normalized to numbers of CFU per milliliter determined from a control culture grown in Sauton’s medium with Tween 80 and plated on 7H10 medium. Data are means ± standard deviations for three independent cultures. The results of all statistical analyses are compared to WT levels unless otherwise indicated. *, P

    Article Snippet: Bacteria were routinely cultured in Middlebrook 7H9 liquid medium (Difco) supplemented with 10% albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD Biosciences) and 0.5% glycerol.

    Techniques: Mutagenesis, Western Blot

    Incomplete repression of eccD 5 transcription has moderate effects on growth. (A to D) Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , eccD 5 Tet-OFF, and Δ pstA1 eccD 5 Tet-OFF strains were inoculated in 7H9 complete medium at an OD 600 of 0.05 and grown at 37°C with aeration. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added at day 0 and day 7 as indicated. Growth was monitored by daily OD 600 measurements (A and C) and by plating serially diluted cultures on 7H10 medium to determine numbers of viable CFU per milliliter on days 0, 3, 6, 9, 12, and 15 (B and D). The key in panel B applies to panels A and B; the key in panel D applies to panels C and D. **, P

    Journal: mBio

    Article Title: Mycobacterium tuberculosis Pst/SenX3-RegX3 Regulates Membrane Vesicle Production Independently of ESX-5 Activity

    doi: 10.1128/mBio.00778-18

    Figure Lengend Snippet: Incomplete repression of eccD 5 transcription has moderate effects on growth. (A to D) Wild-type M. tuberculosis Erdman (WT) and the Δ pstA1 , eccD 5 Tet-OFF, and Δ pstA1 eccD 5 Tet-OFF strains were inoculated in 7H9 complete medium at an OD 600 of 0.05 and grown at 37°C with aeration. Anhydrotetracycline hydrochloride (ATc; 100 ng/ml) was added at day 0 and day 7 as indicated. Growth was monitored by daily OD 600 measurements (A and C) and by plating serially diluted cultures on 7H10 medium to determine numbers of viable CFU per milliliter on days 0, 3, 6, 9, 12, and 15 (B and D). The key in panel B applies to panels A and B; the key in panel D applies to panels C and D. **, P

    Article Snippet: Bacteria were routinely cultured in Middlebrook 7H9 liquid medium (Difco) supplemented with 10% albumin-dextrose-saline (ADS), 0.5% glycerol, and 0.1% Tween 80 or on Middlebrook 7H10 agar medium (Difco) supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD Biosciences) and 0.5% glycerol.

    Techniques:

    In vitro characterization of Δ rv2623 . (A) Growth curves of cultures inoculated (10 6 CFU/ml) into 7H9+10% OADC+0.05% Tween 80 (top) and in minimal Sauton's media (bottom); Erdman (closed boxes, solid line) and Δ rv2623 (open boxes, dashed line) cultures. Error bars represent the standard error of the means; each curve is a combination of at least three independent experiments. (B) Overexpression of Rv2623 in M. smegmatis . The top panel represents serial dilutions (1∶10) of the empty vector pMV261-containing negative control strain and Rv2623-overexpressing strain harboring pMV261:: rv2623 . Diluted stationary phase M. smegmatis culture was spotted (5 µl) onto solid 7H10 media supplemented with 10% OADC and kanamycin (40 µg/mL). Photographs were taken after three days incubation at 37°C. Growth of corresponding strains in liquid medium was assessed based on the time to detection determined using a BD BACTEC 9000MB system (bottom). The various strains were inoculated at 10 4 CFU/ml in triplicates. Data shown are representative of several independent experiments. ***p

    Journal: PLoS Pathogens

    Article Title: Mycobacterium tuberculosis Universal Stress Protein Rv2623 Regulates Bacillary Growth by ATP-Binding: Requirement for Establishing Chronic Persistent Infection

    doi: 10.1371/journal.ppat.1000460

    Figure Lengend Snippet: In vitro characterization of Δ rv2623 . (A) Growth curves of cultures inoculated (10 6 CFU/ml) into 7H9+10% OADC+0.05% Tween 80 (top) and in minimal Sauton's media (bottom); Erdman (closed boxes, solid line) and Δ rv2623 (open boxes, dashed line) cultures. Error bars represent the standard error of the means; each curve is a combination of at least three independent experiments. (B) Overexpression of Rv2623 in M. smegmatis . The top panel represents serial dilutions (1∶10) of the empty vector pMV261-containing negative control strain and Rv2623-overexpressing strain harboring pMV261:: rv2623 . Diluted stationary phase M. smegmatis culture was spotted (5 µl) onto solid 7H10 media supplemented with 10% OADC and kanamycin (40 µg/mL). Photographs were taken after three days incubation at 37°C. Growth of corresponding strains in liquid medium was assessed based on the time to detection determined using a BD BACTEC 9000MB system (bottom). The various strains were inoculated at 10 4 CFU/ml in triplicates. Data shown are representative of several independent experiments. ***p

    Article Snippet: For the determination of the number of colony forming units (CFU) and examination of growth on solid media, Middlebrook 7H10 agar medium (Becton Dickinson) supplemented with 0.5% glycerol and 10% OADC was used.

    Techniques: In Vitro, Over Expression, Plasmid Preparation, Negative Control, 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

    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: A series of modified Middlebrook 7H10 agar media with malachite green concentrations ranging from 2.5 to 2500 mg/L was evaluated using 20 soil samples decontaminated with 3% sodium dodecyl sulfate plus 2% NaOH for 30 min.

    Techniques: Modification, Isolation