m. tuberculosis h37rv Search Results


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  • 99
    ATCC wild type m tuberculosis h37rv
    Wild Type M Tuberculosis H37rv, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Pasteur Institute m tuberculosis h37rv
    Quantitative RT-PCR Expression levels of genes rv3614c , rv3865 , rv3867 , and phoP in duplicate cultures of M. tuberculosis H37Ra and H37Ra:: phoP and <t>H37Rv</t> evaluated by quantitative RT-PCR (qRT-PCR). Genes rv3865 and rv3867 were included as controls as these genes are encoded in the region of difference 1 (RD1) and show similarity to rv3615/14c. The expression level of each gene in each strain is reported as the ratio between the expression level in M. tuberculosis H37Ra culture 1, used as reference. For each strain, values were normalized to the level of 16S rRNA. Note that in M. tuberculosis a single rrs gene is present. Primer and probe sequences of genes rv3614c , rv3865 , rv3867 , phoP , as well as of 16S rRNA are listed in Table S2 .
    M Tuberculosis H37rv, supplied by Pasteur Institute, used in various techniques. Bioz Stars score: 91/100, based on 45 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources m tuberculosis h37rv
    DCP activity against M. tuberculosis <t>H37Rv.</t> PBMCs were plated at a density of 1.5 × 10 6 cells per well in round-bottom 96-well plates and incubated overnight at 37°C. After 2 h of incubation, nonadherent cells were washed off, and the
    M Tuberculosis H37rv, supplied by BEI Resources, used in various techniques. Bioz Stars score: 92/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Glas-Col virulent m tuberculosis h37rv
    DCP activity against M. tuberculosis <t>H37Rv.</t> PBMCs were plated at a density of 1.5 × 10 6 cells per well in round-bottom 96-well plates and incubated overnight at 37°C. After 2 h of incubation, nonadherent cells were washed off, and the
    Virulent M Tuberculosis H37rv, supplied by Glas-Col, used in various techniques. Bioz Stars score: 92/100, based on 18 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources irradiated m tuberculosis h37rv
    DCP activity against M. tuberculosis <t>H37Rv.</t> PBMCs were plated at a density of 1.5 × 10 6 cells per well in round-bottom 96-well plates and incubated overnight at 37°C. After 2 h of incubation, nonadherent cells were washed off, and the
    Irradiated M Tuberculosis H37rv, supplied by BEI Resources, used in various techniques. Bioz Stars score: 93/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore m tuberculosis h37rv tdm
    Genes upregulated in response to M. tuberculosis <t>H37Rv</t> <t>TDM</t> at 2 or 24 h were compared for movement over time. (A) The line graph shows the distinct movement of most of the genes upregulated at 2 h (blue), going back to control levels of expression, while
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    Agilent technologies m tuberculosis h37rv orfs
    Genes upregulated in response to M. tuberculosis <t>H37Rv</t> <t>TDM</t> at 2 or 24 h were compared for movement over time. (A) The line graph shows the distinct movement of most of the genes upregulated at 2 h (blue), going back to control levels of expression, while
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    Bio-Rad m tuberculosis h37rv dna
    Genes upregulated in response to M. tuberculosis <t>H37Rv</t> <t>TDM</t> at 2 or 24 h were compared for movement over time. (A) The line graph shows the distinct movement of most of the genes upregulated at 2 h (blue), going back to control levels of expression, while
    M Tuberculosis H37rv Dna, supplied by Bio-Rad, used in various techniques. Bioz Stars score: 86/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Glas-Col m tuberculosis h37rv
    Characteristics of the Mycobacterium tuberculosis P hsp60 TK strain. A. Equal numbers of M. tuberculosis <t>H37Rv</t> (wild-type) or M. tuberculosis P hsp60 TK strains were incubated with 1 µCi/ml of [ 125 I]FIAU at 37°C in Middlebrook 7H9 liquid broth for 6 and 24 hours. At each specified time-point equal aliquots were withdrawn from the cultures and washed 3 times to remove the media. Each pellet was resuspended in PBS in 1.5 ml Eppendorf tubes and disinfected with Lysol overnight. The activity for each Eppendorf was measured using a gamma counter. M. tuberculosis P hsp60 TK strain actively accumulates [ 125 I]FIAU in vitro . Mean uptake activity is 6086 (±536) and 8011 (±3233) counts per minute (cpm) at 6 and 24 hours for the M. tuberculosis P hsp60 TK strain which is significantly more than 549 (±50) and 615 (±260) cpm for the wild-type parent strain (p
    M Tuberculosis H37rv, supplied by Glas-Col, used in various techniques. Bioz Stars score: 94/100, based on 117 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    MiddleBrook Pharmaceuticals m tuberculosis h37rv
    Ability of BCG vaccines to protect guinea pigs from hematogenous spread following aerogenic challenge with M. bovis or M. tuberculosis . Data are numbers of CFU per milliliter recovered from the spleens of animals 4 weeks after challenge via the aerogenic route with M. bovis strain 1692/96 or M. tuberculosis strain <t>H37Rv</t> (limit of detection for both = 5 CFU/ml) at a retained dose of 10 organisms. Five weeks before challenge, animals were vaccinated with 5 × 10 4 CFU of BCG Pasteur, a BCG methionine auxotroph (Met − ), or a BCG leucine auxotroph (Leu − ).
    M Tuberculosis H37rv, supplied by MiddleBrook Pharmaceuticals, used in various techniques. Bioz Stars score: 97/100, based on 426 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Advanced Biotechnologies Inc m tuberculosis h37rv international standard
    Linearity of IS6110 (ISP) and senX3 qPCR. M. tuberculosis <t>H37Rv</t> commercial standard (A,B) and M. bovis BCG genomic DNA (C,D) .
    M Tuberculosis H37rv International Standard, supplied by Advanced Biotechnologies Inc, used in various techniques. Bioz Stars score: 94/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources growth conditions m tuberculosis h37rv
    Linearity of IS6110 (ISP) and senX3 qPCR. M. tuberculosis <t>H37Rv</t> commercial standard (A,B) and M. bovis BCG genomic DNA (C,D) .
    Growth Conditions M Tuberculosis H37rv, supplied by BEI Resources, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Agilent technologies m tuberculosis h37rv
    Correspondence analysis (CA) of all samples from 5 time-points post-infection of mouse macrophages with wild-type M. tuberculosis <t>H37Rv</t> (Rv) or M. tuberculosis H37Rv Δ-mce1 (Yk). Samples from the ‘early’ time-points post H37Rv-infection (15 min 30 min and 60 min) are colored yellow/orange, and samples from the ‘late’ time-points post H37Rv-infection (4 hrs and 10 hrs) are colored red/brown. Samples from the ‘early’ time-points post Δ-mce1-H37Rv infection (15 min, 30 min, and 60 min) are colored light green, and samples from the ‘late’ time-points post Δ-mce1-H37Rv -infection (4 hrs and 10 hrs]) are dark green/black. The uninfected samples (U) are colored purple, and the pooled common reference samples are colored blue.
    M Tuberculosis H37rv, supplied by Agilent technologies, used in various techniques. Bioz Stars score: 86/100, based on 20 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    GenScript m tuberculosis h37rv
    Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis <t>H37Rv</t> intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).
    M Tuberculosis H37rv, supplied by GenScript, used in various techniques. Bioz Stars score: 91/100, based on 22 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Thermo Fisher m tuberculosis h37rv
    Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis <t>H37Rv</t> intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).
    M Tuberculosis H37rv, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 93/100, based on 186 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    HUYGEN m tuberculosis h37rv
    Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis <t>H37Rv</t> intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).
    M Tuberculosis H37rv, supplied by HUYGEN, used in various techniques. Bioz Stars score: 85/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Glas-Col aerosolized m tuberculosis h37rv
    Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis <t>H37Rv</t> intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).
    Aerosolized M Tuberculosis H37rv, supplied by Glas-Col, used in various techniques. Bioz Stars score: 90/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources m tuberculosis h37rv lysate
    Deletion of nuoG in BCG improves protection of mice against TB. (A) Schematic design of protection studies. (B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), or BCG Δ nuoG (▼) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain <t>H37Rv.</t> Bacterial burdens of lungs and spleen were determined at 30 and 180 days postchallenge by plating organ homogenates on agar. Shown are means ± standard deviations. Results are representative of two experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test ( n = 8). *, P
    M Tuberculosis H37rv Lysate, supplied by BEI Resources, used in various techniques. Bioz Stars score: 93/100, based on 14 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Glas-Col m tuberculosis h37rv mouse
    Deletion of nuoG in BCG improves protection of mice against TB. (A) Schematic design of protection studies. (B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), or BCG Δ nuoG (▼) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain <t>H37Rv.</t> Bacterial burdens of lungs and spleen were determined at 30 and 180 days postchallenge by plating organ homogenates on agar. Shown are means ± standard deviations. Results are representative of two experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test ( n = 8). *, P
    M Tuberculosis H37rv Mouse, supplied by Glas-Col, used in various techniques. Bioz Stars score: 87/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC m tuberculosis h37rv
    TAC-HRM of rrs (amplicon 3) and gyrB (amplicon 2) gene segments. Singleplex PCR mixtures with primers and probes were loaded into empty microfluidic cards (1 sample per port, yielding 48 reaction mixtures per sample). Results are shown for probe-based detection (A and E), for amplification with SYTO9 (B and F), and for melt curve and difference curve analysis (C and D, G and H). The aligned melt curves show melt temperature shifts versus those for the M. tuberculosis <t>H37Rv</t> wild-type control based on nucleotide changes (leftward variation indicates a lower melt temperature and rightward variation a higher melt temperature). The difference plot uses the same data but plots the negative first derivative (− dF / dt ) on the y axis. The HRM software automatically labels samples that are variants from the wild-type control with a different color. Underlining indicates nucleotides that changed. The rrs (amplicon 3) primer/probe results show the amplification curve obtained with the rrs -A(1401)G-specific probe for a mutant isolate, while the results for M. tuberculosis H37Rv were negative. Both the mutant isolate and H37Rv amplified with the primers using SYTO9, and the HRM result reinforced the probe result as being a mutant/variant sample. The gyrB (amplicon 2) primer/probe results demonstrated the benefit of combining the probe with HRM, since the specific probe (E) detected the Asp461His (GAC→CAC) transversion, while HRM would not (F, G, and H); however, HRM detected the rare mutations Asp461Ala and Asp461Asn.
    M Tuberculosis H37rv, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 1284 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    91
    Pasteur Institute m tuberculosis h37rv strain
    TAC-HRM of rrs (amplicon 3) and gyrB (amplicon 2) gene segments. Singleplex PCR mixtures with primers and probes were loaded into empty microfluidic cards (1 sample per port, yielding 48 reaction mixtures per sample). Results are shown for probe-based detection (A and E), for amplification with SYTO9 (B and F), and for melt curve and difference curve analysis (C and D, G and H). The aligned melt curves show melt temperature shifts versus those for the M. tuberculosis <t>H37Rv</t> wild-type control based on nucleotide changes (leftward variation indicates a lower melt temperature and rightward variation a higher melt temperature). The difference plot uses the same data but plots the negative first derivative (− dF / dt ) on the y axis. The HRM software automatically labels samples that are variants from the wild-type control with a different color. Underlining indicates nucleotides that changed. The rrs (amplicon 3) primer/probe results show the amplification curve obtained with the rrs -A(1401)G-specific probe for a mutant isolate, while the results for M. tuberculosis H37Rv were negative. Both the mutant isolate and H37Rv amplified with the primers using SYTO9, and the HRM result reinforced the probe result as being a mutant/variant sample. The gyrB (amplicon 2) primer/probe results demonstrated the benefit of combining the probe with HRM, since the specific probe (E) detected the Asp461His (GAC→CAC) transversion, while HRM would not (F, G, and H); however, HRM detected the rare mutations Asp461Ala and Asp461Asn.
    M Tuberculosis H37rv Strain, supplied by Pasteur Institute, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources m tuberculosis h37rv nr 13648
    TAC-HRM of rrs (amplicon 3) and gyrB (amplicon 2) gene segments. Singleplex PCR mixtures with primers and probes were loaded into empty microfluidic cards (1 sample per port, yielding 48 reaction mixtures per sample). Results are shown for probe-based detection (A and E), for amplification with SYTO9 (B and F), and for melt curve and difference curve analysis (C and D, G and H). The aligned melt curves show melt temperature shifts versus those for the M. tuberculosis <t>H37Rv</t> wild-type control based on nucleotide changes (leftward variation indicates a lower melt temperature and rightward variation a higher melt temperature). The difference plot uses the same data but plots the negative first derivative (− dF / dt ) on the y axis. The HRM software automatically labels samples that are variants from the wild-type control with a different color. Underlining indicates nucleotides that changed. The rrs (amplicon 3) primer/probe results show the amplification curve obtained with the rrs -A(1401)G-specific probe for a mutant isolate, while the results for M. tuberculosis H37Rv were negative. Both the mutant isolate and H37Rv amplified with the primers using SYTO9, and the HRM result reinforced the probe result as being a mutant/variant sample. The gyrB (amplicon 2) primer/probe results demonstrated the benefit of combining the probe with HRM, since the specific probe (E) detected the Asp461His (GAC→CAC) transversion, while HRM would not (F, G, and H); however, HRM detected the rare mutations Asp461Ala and Asp461Asn.
    M Tuberculosis H37rv Nr 13648, supplied by BEI Resources, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC virulent m tuberculosis h37rv
    Kaplan-Meier plot of the survival of guinea pigs after low dose aerosol infection with virulent M. tuberculosis <t>H37Rv.</t> Guinea pigs were inoculated with saline, BCG, CFP+MPL/DDA, Ag85A+MPL/DDA or MPL/DDA and rested for 10 weeks prior to infection. All
    Virulent M Tuberculosis H37rv, supplied by ATCC, used in various techniques. Bioz Stars score: 99/100, based on 23 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    BEI Resources m tuberculosis h37rv derived culture filtrate protein
    Vaccination with BCG or BCGΔBCG149c does not change immune cells activation in response to CFP after infection. C57BL/6 mice ( n = 5) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis <t>H37Rv</t> by aerosol route, and 6 months post-infection, lysates from (A) spleen, (B) lymph nodes, and (C) lungs, and stimulated or not with M. tuberculosis H37Rv CFP. Data (SFUs/2 × 10 5 cells) from individual samples are shown, with the mean value for each group indicated by a horizontal line. Data were analyzed for statistical significance between the groups by ANOVA, and we found variable response from animal to animal, with no statistical significance between vaccinated and non-vaccinated mice.
    M Tuberculosis H37rv Derived Culture Filtrate Protein, supplied by BEI Resources, used in various techniques. Bioz Stars score: 94/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    AstraZeneca m tuberculosis h37rv genomic dna
    Vaccination with BCG or BCGΔBCG149c does not change immune cells activation in response to CFP after infection. C57BL/6 mice ( n = 5) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis <t>H37Rv</t> by aerosol route, and 6 months post-infection, lysates from (A) spleen, (B) lymph nodes, and (C) lungs, and stimulated or not with M. tuberculosis H37Rv CFP. Data (SFUs/2 × 10 5 cells) from individual samples are shown, with the mean value for each group indicated by a horizontal line. Data were analyzed for statistical significance between the groups by ANOVA, and we found variable response from animal to animal, with no statistical significance between vaccinated and non-vaccinated mice.
    M Tuberculosis H37rv Genomic Dna, supplied by AstraZeneca, used in various techniques. Bioz Stars score: 86/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Millipore m tuberculosis h37rv sigma orf
    Vaccination with BCG or BCGΔBCG149c does not change immune cells activation in response to CFP after infection. C57BL/6 mice ( n = 5) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis <t>H37Rv</t> by aerosol route, and 6 months post-infection, lysates from (A) spleen, (B) lymph nodes, and (C) lungs, and stimulated or not with M. tuberculosis H37Rv CFP. Data (SFUs/2 × 10 5 cells) from individual samples are shown, with the mean value for each group indicated by a horizontal line. Data were analyzed for statistical significance between the groups by ANOVA, and we found variable response from animal to animal, with no statistical significance between vaccinated and non-vaccinated mice.
    M Tuberculosis H37rv Sigma Orf, supplied by Millipore, 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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    ATCC m tuberculosis h37rv 25618
    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis <t>H37Rv-MA</t> or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).
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    tebu-bio sa m tuberculosis h37rv reference strain
    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis <t>H37Rv-MA</t> or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).
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    Thermo Fisher m tuberculosis h37rv genomic dna
    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis <t>H37Rv-MA</t> or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).
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    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis <t>H37Rv-MA</t> or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).
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    ATCC virulent m tuberculosis h37rv strain
    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis <t>H37Rv-MA</t> or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).
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    Image Search Results


    Quantitative RT-PCR Expression levels of genes rv3614c , rv3865 , rv3867 , and phoP in duplicate cultures of M. tuberculosis H37Ra and H37Ra:: phoP and H37Rv evaluated by quantitative RT-PCR (qRT-PCR). Genes rv3865 and rv3867 were included as controls as these genes are encoded in the region of difference 1 (RD1) and show similarity to rv3615/14c. The expression level of each gene in each strain is reported as the ratio between the expression level in M. tuberculosis H37Ra culture 1, used as reference. For each strain, values were normalized to the level of 16S rRNA. Note that in M. tuberculosis a single rrs gene is present. Primer and probe sequences of genes rv3614c , rv3865 , rv3867 , phoP , as well as of 16S rRNA are listed in Table S2 .

    Journal: PLoS Pathogens

    Article Title: Control of M. tuberculosis ESAT-6 Secretion and Specific T Cell Recognition by PhoP

    doi: 10.1371/journal.ppat.0040033

    Figure Lengend Snippet: Quantitative RT-PCR Expression levels of genes rv3614c , rv3865 , rv3867 , and phoP in duplicate cultures of M. tuberculosis H37Ra and H37Ra:: phoP and H37Rv evaluated by quantitative RT-PCR (qRT-PCR). Genes rv3865 and rv3867 were included as controls as these genes are encoded in the region of difference 1 (RD1) and show similarity to rv3615/14c. The expression level of each gene in each strain is reported as the ratio between the expression level in M. tuberculosis H37Ra culture 1, used as reference. For each strain, values were normalized to the level of 16S rRNA. Note that in M. tuberculosis a single rrs gene is present. Primer and probe sequences of genes rv3614c , rv3865 , rv3867 , phoP , as well as of 16S rRNA are listed in Table S2 .

    Article Snippet: M. tuberculosis H37Ra corresponding to ATCC 25177 and M. tuberculosis H37Rv were taken from stocks held at the Pasteur Institute.

    Techniques: Quantitative RT-PCR, Expressing

    Secretion Analysis In vitro expression and secretion of ESAT-6 from M. tuberculosis H37Rv, H37Ra, MT103 phoP ko (SO2), and recombinant H37Ra complemented with integrating cosmids carrying genes phoP or rpsL from H37Rv. Total protein concentrations were determined by using Bio-Rad protein assay, and 15-μg samples were subjected to SDS-PAGE. Detection was carried out by using monoclonal anti-ESAT-6 antibody Hyb 76–8a.

    Journal: PLoS Pathogens

    Article Title: Control of M. tuberculosis ESAT-6 Secretion and Specific T Cell Recognition by PhoP

    doi: 10.1371/journal.ppat.0040033

    Figure Lengend Snippet: Secretion Analysis In vitro expression and secretion of ESAT-6 from M. tuberculosis H37Rv, H37Ra, MT103 phoP ko (SO2), and recombinant H37Ra complemented with integrating cosmids carrying genes phoP or rpsL from H37Rv. Total protein concentrations were determined by using Bio-Rad protein assay, and 15-μg samples were subjected to SDS-PAGE. Detection was carried out by using monoclonal anti-ESAT-6 antibody Hyb 76–8a.

    Article Snippet: M. tuberculosis H37Ra corresponding to ATCC 25177 and M. tuberculosis H37Rv were taken from stocks held at the Pasteur Institute.

    Techniques: In Vitro, Expressing, Recombinant, SDS Page

    Antigen-Specific IL-2 Production of T Cell Hybridomas Analysis of IL-2 secretion by anti-ESAT-6 or anti-Ag85A T cell hybridomas in response to recognition of antigen presented by bone marrow–derived dendritic cells (BM-DC) incubated with homologous or control peptides or infected with H37Rv, H37Ra or recombinant H37Ra strains.

    Journal: PLoS Pathogens

    Article Title: Control of M. tuberculosis ESAT-6 Secretion and Specific T Cell Recognition by PhoP

    doi: 10.1371/journal.ppat.0040033

    Figure Lengend Snippet: Antigen-Specific IL-2 Production of T Cell Hybridomas Analysis of IL-2 secretion by anti-ESAT-6 or anti-Ag85A T cell hybridomas in response to recognition of antigen presented by bone marrow–derived dendritic cells (BM-DC) incubated with homologous or control peptides or infected with H37Rv, H37Ra or recombinant H37Ra strains.

    Article Snippet: M. tuberculosis H37Ra corresponding to ATCC 25177 and M. tuberculosis H37Rv were taken from stocks held at the Pasteur Institute.

    Techniques: Derivative Assay, Incubation, Infection, Recombinant

    Macrophage Infection Studies Bone marrow–derived murine macrophages (BMM) were infected with M. tuberculosis H37Ra, H37Ra:: fadE5 , H37Ra:: phoP , and H37Rv at a MOI of ca. 1:1 and 10:1 bacteria per cell (A and B, respectively). The figure shows the means and standard deviations of CFU ratio values (CFU at days 1, 4, and 7 relative to CFU at 4 h) obtained in a representative experiment performed in quadruplicate. The significant difference levels in growth characteristics between H37Ra and other strains (H37Ra:: phoP and H37Rv) were determined by analysis of variance (ANOVA) (* p

    Journal: PLoS Pathogens

    Article Title: Control of M. tuberculosis ESAT-6 Secretion and Specific T Cell Recognition by PhoP

    doi: 10.1371/journal.ppat.0040033

    Figure Lengend Snippet: Macrophage Infection Studies Bone marrow–derived murine macrophages (BMM) were infected with M. tuberculosis H37Ra, H37Ra:: fadE5 , H37Ra:: phoP , and H37Rv at a MOI of ca. 1:1 and 10:1 bacteria per cell (A and B, respectively). The figure shows the means and standard deviations of CFU ratio values (CFU at days 1, 4, and 7 relative to CFU at 4 h) obtained in a representative experiment performed in quadruplicate. The significant difference levels in growth characteristics between H37Ra and other strains (H37Ra:: phoP and H37Rv) were determined by analysis of variance (ANOVA) (* p

    Article Snippet: M. tuberculosis H37Ra corresponding to ATCC 25177 and M. tuberculosis H37Rv were taken from stocks held at the Pasteur Institute.

    Techniques: Infection, Derivative Assay

    QTL linkage analysis of TB severity following infection of mice with M. tuberculosis H37Rv: LOD score plots for gender-adjusted postinfection relative body weight loss and survival time from analyses of BC1 (A and D) and F2 (B, C, and E) animals. The LOD score peaks between D9Mit23 and D9M142 in panel A were not designated since they were not supported by a marker.

    Journal: Infection and Immunity

    Article Title: Multigenic Control of Disease Severity after Virulent Mycobacterium tuberculosis Infection in Mice

    doi: 10.1128/IAI.71.1.126-131.2003

    Figure Lengend Snippet: QTL linkage analysis of TB severity following infection of mice with M. tuberculosis H37Rv: LOD score plots for gender-adjusted postinfection relative body weight loss and survival time from analyses of BC1 (A and D) and F2 (B, C, and E) animals. The LOD score peaks between D9Mit23 and D9M142 in panel A were not designated since they were not supported by a marker.

    Article Snippet: M. tuberculosis H37Rv Pasteur (a kind gift from Gilles Marchale, Pasteur Institute, Paris, France) was grown and stored exactly as previously described ( , ).

    Techniques: Infection, Mouse Assay, Marker

    DCP activity against M. tuberculosis H37Rv. PBMCs were plated at a density of 1.5 × 10 6 cells per well in round-bottom 96-well plates and incubated overnight at 37°C. After 2 h of incubation, nonadherent cells were washed off, and the

    Journal: Infection and Immunity

    Article Title: Dipterinyl Calcium Pentahydrate Inhibits Intracellular Mycobacterial Growth in Human Monocytes via the C-C Chemokine MIP-1? and Nitric Oxide

    doi: 10.1128/IAI.01393-12

    Figure Lengend Snippet: DCP activity against M. tuberculosis H37Rv. PBMCs were plated at a density of 1.5 × 10 6 cells per well in round-bottom 96-well plates and incubated overnight at 37°C. After 2 h of incubation, nonadherent cells were washed off, and the

    Article Snippet: M. tuberculosis H37Rv (catalog number NR-123) was obtained from BEI Resources, Manassas, VA. Before being used for infection, bacteria were thawed and sonicated for l min in a cup sonicator (W385; Heat Systems Ultrasonics, Farmingdale, NY) to obtain a single-cell suspension and diluted appropriately in antibiotic-free complete RPMI 1640 medium.

    Techniques: Activity Assay, Incubation

    Genes upregulated in response to M. tuberculosis H37Rv TDM at 2 or 24 h were compared for movement over time. (A) The line graph shows the distinct movement of most of the genes upregulated at 2 h (blue), going back to control levels of expression, while

    Journal: Infection and Immunity

    Article Title: Mycobacterial Trehalose Dimycolate Reprograms Macrophage Global Gene Expression and Activates Matrix Metalloproteinases

    doi: 10.1128/IAI.00906-12

    Figure Lengend Snippet: Genes upregulated in response to M. tuberculosis H37Rv TDM at 2 or 24 h were compared for movement over time. (A) The line graph shows the distinct movement of most of the genes upregulated at 2 h (blue), going back to control levels of expression, while

    Article Snippet: One hundred micrograms of M. tuberculosis H37Rv TDM (Sigma-Aldrich) was used to coat solvent-resistant tubes, to which were added 2 × 103 90-μm-diameter or 3 × 106 10-μm-diameter washed polystyrene microspheres suspended in 1 ml of PBS.

    Techniques: Expressing

    Most of the macrophage genes regulated in response to TDM are MyD88 dependent. BMMΦ genes that were shown to be up- or downregulated in response to H37Rv TDM after normalization against PG were analyzed in the microarray data acquired by the comparison

    Journal: Infection and Immunity

    Article Title: Mycobacterial Trehalose Dimycolate Reprograms Macrophage Global Gene Expression and Activates Matrix Metalloproteinases

    doi: 10.1128/IAI.00906-12

    Figure Lengend Snippet: Most of the macrophage genes regulated in response to TDM are MyD88 dependent. BMMΦ genes that were shown to be up- or downregulated in response to H37Rv TDM after normalization against PG were analyzed in the microarray data acquired by the comparison

    Article Snippet: One hundred micrograms of M. tuberculosis H37Rv TDM (Sigma-Aldrich) was used to coat solvent-resistant tubes, to which were added 2 × 103 90-μm-diameter or 3 × 106 10-μm-diameter washed polystyrene microspheres suspended in 1 ml of PBS.

    Techniques: Microarray

    Macrophage gene regulation in response to TDM. BMMΦ from C57BL/6 mice were stimulated with 90-μm-diameter microspheres coated with TDM purified from M. tuberculosis strain H37Rv for either 2 (A) or 24 (B) hours. Dots represent individual

    Journal: Infection and Immunity

    Article Title: Mycobacterial Trehalose Dimycolate Reprograms Macrophage Global Gene Expression and Activates Matrix Metalloproteinases

    doi: 10.1128/IAI.00906-12

    Figure Lengend Snippet: Macrophage gene regulation in response to TDM. BMMΦ from C57BL/6 mice were stimulated with 90-μm-diameter microspheres coated with TDM purified from M. tuberculosis strain H37Rv for either 2 (A) or 24 (B) hours. Dots represent individual

    Article Snippet: One hundred micrograms of M. tuberculosis H37Rv TDM (Sigma-Aldrich) was used to coat solvent-resistant tubes, to which were added 2 × 103 90-μm-diameter or 3 × 106 10-μm-diameter washed polystyrene microspheres suspended in 1 ml of PBS.

    Techniques: Mouse Assay, Purification

    Characteristics of the Mycobacterium tuberculosis P hsp60 TK strain. A. Equal numbers of M. tuberculosis H37Rv (wild-type) or M. tuberculosis P hsp60 TK strains were incubated with 1 µCi/ml of [ 125 I]FIAU at 37°C in Middlebrook 7H9 liquid broth for 6 and 24 hours. At each specified time-point equal aliquots were withdrawn from the cultures and washed 3 times to remove the media. Each pellet was resuspended in PBS in 1.5 ml Eppendorf tubes and disinfected with Lysol overnight. The activity for each Eppendorf was measured using a gamma counter. M. tuberculosis P hsp60 TK strain actively accumulates [ 125 I]FIAU in vitro . Mean uptake activity is 6086 (±536) and 8011 (±3233) counts per minute (cpm) at 6 and 24 hours for the M. tuberculosis P hsp60 TK strain which is significantly more than 549 (±50) and 615 (±260) cpm for the wild-type parent strain (p

    Journal: PLoS ONE

    Article Title: Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals

    doi: 10.1371/journal.pone.0006297

    Figure Lengend Snippet: Characteristics of the Mycobacterium tuberculosis P hsp60 TK strain. A. Equal numbers of M. tuberculosis H37Rv (wild-type) or M. tuberculosis P hsp60 TK strains were incubated with 1 µCi/ml of [ 125 I]FIAU at 37°C in Middlebrook 7H9 liquid broth for 6 and 24 hours. At each specified time-point equal aliquots were withdrawn from the cultures and washed 3 times to remove the media. Each pellet was resuspended in PBS in 1.5 ml Eppendorf tubes and disinfected with Lysol overnight. The activity for each Eppendorf was measured using a gamma counter. M. tuberculosis P hsp60 TK strain actively accumulates [ 125 I]FIAU in vitro . Mean uptake activity is 6086 (±536) and 8011 (±3233) counts per minute (cpm) at 6 and 24 hours for the M. tuberculosis P hsp60 TK strain which is significantly more than 549 (±50) and 615 (±260) cpm for the wild-type parent strain (p

    Article Snippet: C3HeB/FeJ mice were low-dose aerosol infected with equal numbers of either M. tuberculosis H37Rv or M. tuberculosis Phsp60 TK strains using the Inhalation Exposure System (Glas-Col).

    Techniques: Incubation, Activity Assay, In Vitro

    [ 125 I]FIAU-SPECT can detect and localize Mycobacterium tuberculosis P hsp60 TK strain in situ . M. tuberculosis H37Rv wild-type or P hsp60 TK strain (∼10 8 CFU) were inoculated into either thighs of a BALB/c mouse. 1 mCi of [ 125 I]FIAU was injected via a tail vein injection. Panels A and B show the fused SPECT and CT images 3 and 12 hours after the [ 125 I]FIAU injection. Signal is detected at both time-points in the thigh inoculated with M. tuberculosis P hsp60 TK strain but not in the thigh inoculated with the wild-type strain. As expected, SPECT signal is detected in several tissues [liver/gall bladder (L), stomach (St), gastrointestinal tract (GI), urinary bladder (UB)] that either metabolize or excrete FIAU or its iodinated derivatives.

    Journal: PLoS ONE

    Article Title: Bacterial Thymidine Kinase as a Non-Invasive Imaging Reporter for Mycobacterium tuberculosis in Live Animals

    doi: 10.1371/journal.pone.0006297

    Figure Lengend Snippet: [ 125 I]FIAU-SPECT can detect and localize Mycobacterium tuberculosis P hsp60 TK strain in situ . M. tuberculosis H37Rv wild-type or P hsp60 TK strain (∼10 8 CFU) were inoculated into either thighs of a BALB/c mouse. 1 mCi of [ 125 I]FIAU was injected via a tail vein injection. Panels A and B show the fused SPECT and CT images 3 and 12 hours after the [ 125 I]FIAU injection. Signal is detected at both time-points in the thigh inoculated with M. tuberculosis P hsp60 TK strain but not in the thigh inoculated with the wild-type strain. As expected, SPECT signal is detected in several tissues [liver/gall bladder (L), stomach (St), gastrointestinal tract (GI), urinary bladder (UB)] that either metabolize or excrete FIAU or its iodinated derivatives.

    Article Snippet: C3HeB/FeJ mice were low-dose aerosol infected with equal numbers of either M. tuberculosis H37Rv or M. tuberculosis Phsp60 TK strains using the Inhalation Exposure System (Glas-Col).

    Techniques: Single Photon Emission Computed Tomography, In Situ, Injection

    Ability of BCG vaccines to protect guinea pigs from hematogenous spread following aerogenic challenge with M. bovis or M. tuberculosis . Data are numbers of CFU per milliliter recovered from the spleens of animals 4 weeks after challenge via the aerogenic route with M. bovis strain 1692/96 or M. tuberculosis strain H37Rv (limit of detection for both = 5 CFU/ml) at a retained dose of 10 organisms. Five weeks before challenge, animals were vaccinated with 5 × 10 4 CFU of BCG Pasteur, a BCG methionine auxotroph (Met − ), or a BCG leucine auxotroph (Leu − ).

    Journal: Infection and Immunity

    Article Title: Identification of a Mycobacterium bovis BCG Auxotrophic Mutant That Protects Guinea Pigs against M. bovis and Hematogenous Spread of Mycobacterium tuberculosis without Sensitization to Tuberculin

    doi:

    Figure Lengend Snippet: Ability of BCG vaccines to protect guinea pigs from hematogenous spread following aerogenic challenge with M. bovis or M. tuberculosis . Data are numbers of CFU per milliliter recovered from the spleens of animals 4 weeks after challenge via the aerogenic route with M. bovis strain 1692/96 or M. tuberculosis strain H37Rv (limit of detection for both = 5 CFU/ml) at a retained dose of 10 organisms. Five weeks before challenge, animals were vaccinated with 5 × 10 4 CFU of BCG Pasteur, a BCG methionine auxotroph (Met − ), or a BCG leucine auxotroph (Leu − ).

    Article Snippet: A seed stock of M. tuberculosis H37Rv was plated and grown on Middlebrook 7H10 agar containing 0.2% (vol/vol) glycerol and 10% (vol/vol) Middlebrook OADC enrichment for 3 weeks at 37°C and then harvested into sterile deionized water.

    Techniques:

    Ability of BCG vaccines to protect guinea pigs from pulmonary disease. Disease severity score based on the number and size of lesions, together with the presence of caseation, on the dorsal surface of formalin-fixed lungs removed 4 weeks after challenge via the aerogenic route with M. bovis strain 1692/96 or M. tuberculosis strain H37Rv at a retained dose of 10 organisms. Five weeks before challenge, animals were vaccinated with 5 × 10 4 CFU of BCG Pasteur, a BCG methionine auxotroph (Met − ), or a BCG leucine auxotroph (Leu − ).

    Journal: Infection and Immunity

    Article Title: Identification of a Mycobacterium bovis BCG Auxotrophic Mutant That Protects Guinea Pigs against M. bovis and Hematogenous Spread of Mycobacterium tuberculosis without Sensitization to Tuberculin

    doi:

    Figure Lengend Snippet: Ability of BCG vaccines to protect guinea pigs from pulmonary disease. Disease severity score based on the number and size of lesions, together with the presence of caseation, on the dorsal surface of formalin-fixed lungs removed 4 weeks after challenge via the aerogenic route with M. bovis strain 1692/96 or M. tuberculosis strain H37Rv at a retained dose of 10 organisms. Five weeks before challenge, animals were vaccinated with 5 × 10 4 CFU of BCG Pasteur, a BCG methionine auxotroph (Met − ), or a BCG leucine auxotroph (Leu − ).

    Article Snippet: A seed stock of M. tuberculosis H37Rv was plated and grown on Middlebrook 7H10 agar containing 0.2% (vol/vol) glycerol and 10% (vol/vol) Middlebrook OADC enrichment for 3 weeks at 37°C and then harvested into sterile deionized water.

    Techniques:

    PLG synthesis by M. tuberculosis H37Rv in different growth phases, under different growth conditions, and in biofilms.

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: PLG synthesis by M. tuberculosis H37Rv in different growth phases, under different growth conditions, and in biofilms.

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques:

    PLG contents of M. tuberculosis H37Rv cells from different phases of growth. *, P

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: PLG contents of M. tuberculosis H37Rv cells from different phases of growth. *, P

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques:

    Growth and persistence of mycobacterial strains in lung and spleen of BALB/c mice. (A) Replication of H37Rv, ΔRv0574c, and ΔRv0574c-comp strains in lungs of BALB/c mice. (B) Replication of the strains in spleens of BALB/c mice. #, P

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: Growth and persistence of mycobacterial strains in lung and spleen of BALB/c mice. (A) Replication of H37Rv, ΔRv0574c, and ΔRv0574c-comp strains in lungs of BALB/c mice. (B) Replication of the strains in spleens of BALB/c mice. #, P

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques: Mouse Assay

    Growth profiles of mycobacterial strains in broth culture. H37Rv, ΔRv0574c, and ΔRv0574c-comp strains were inoculated at an initial optical density of 0.02 in enriched 7H9 medium. (A) OD 600 . (B) Log CFU/ml of cultures.

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: Growth profiles of mycobacterial strains in broth culture. H37Rv, ΔRv0574c, and ΔRv0574c-comp strains were inoculated at an initial optical density of 0.02 in enriched 7H9 medium. (A) OD 600 . (B) Log CFU/ml of cultures.

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques:

    Real-time PCR to analyze Rv0574c expression in M. tuberculosis H37Rv. (A) Expression in different growth phases. Fold change was calculated in late-log- and stationary-phase cells, taking early log expression as 1, and is indicated by vertical bars; the

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: Real-time PCR to analyze Rv0574c expression in M. tuberculosis H37Rv. (A) Expression in different growth phases. Fold change was calculated in late-log- and stationary-phase cells, taking early log expression as 1, and is indicated by vertical bars; the

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques: Real-time Polymerase Chain Reaction, Expressing

    Growth of ΔRv0574c strain in human THP-1 macrophages. Differentiated THP-1 cells were infected with H37Rv, ΔRv0574c, and ΔRv0574c-comp strains at an MOI of 10. Infected cells were lysed at the indicated time points for CFU counts.

    Journal: Infection and Immunity

    Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

    doi: 10.1128/IAI.02274-14

    Figure Lengend Snippet: Growth of ΔRv0574c strain in human THP-1 macrophages. Differentiated THP-1 cells were infected with H37Rv, ΔRv0574c, and ΔRv0574c-comp strains at an MOI of 10. Infected cells were lysed at the indicated time points for CFU counts.

    Article Snippet: M. tuberculosis H37Rv strains were grown on Middlebrook 7H10 or 7H11 agar (Difco) containing 10% (vol/vol) oleic acid, albumin, dextrose, catalase (OADC) (Becton Dickinson); 0.5% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80 or in 7H9 broth (Difco) supplemented with 10% (vol/vol) albumin, dextrose, catalase (ADC) (Becton Dickinson); 0.2% (vol/vol) glycerol; and 0.05% (vol/vol) Tween 80, at 37°C with shaking at 180 rpm.

    Techniques: Infection

    Linearity of IS6110 (ISP) and senX3 qPCR. M. tuberculosis H37Rv commercial standard (A,B) and M. bovis BCG genomic DNA (C,D) .

    Journal: Frontiers in Microbiology

    Article Title: Optimized Lysis-Extraction Method Combined With IS6110-Amplification for Detection of Mycobacterium tuberculosis in Paucibacillary Sputum Specimens

    doi: 10.3389/fmicb.2018.02224

    Figure Lengend Snippet: Linearity of IS6110 (ISP) and senX3 qPCR. M. tuberculosis H37Rv commercial standard (A,B) and M. bovis BCG genomic DNA (C,D) .

    Article Snippet: M. tuberculosis H37Rv international standard (Advanced Biotechnologies Inc., Eldersburg, MD, United States) was used to generate an accurate standard curve of PCR using concentrations. senX3 PCR was performed, from other primers and probe designed to target the senX3-regX3 intergenic region, as previously described ( ).

    Techniques: Real-time Polymerase Chain Reaction

    Correspondence analysis (CA) of all samples from 5 time-points post-infection of mouse macrophages with wild-type M. tuberculosis H37Rv (Rv) or M. tuberculosis H37Rv Δ-mce1 (Yk). Samples from the ‘early’ time-points post H37Rv-infection (15 min 30 min and 60 min) are colored yellow/orange, and samples from the ‘late’ time-points post H37Rv-infection (4 hrs and 10 hrs) are colored red/brown. Samples from the ‘early’ time-points post Δ-mce1-H37Rv infection (15 min, 30 min, and 60 min) are colored light green, and samples from the ‘late’ time-points post Δ-mce1-H37Rv -infection (4 hrs and 10 hrs]) are dark green/black. The uninfected samples (U) are colored purple, and the pooled common reference samples are colored blue.

    Journal: PLoS ONE

    Article Title: Modulation of Transcriptional and Inflammatory Responses in Murine Macrophages by the Mycobacterium tuberculosis Mammalian Cell Entry (Mce) 1 Complex

    doi: 10.1371/journal.pone.0026295

    Figure Lengend Snippet: Correspondence analysis (CA) of all samples from 5 time-points post-infection of mouse macrophages with wild-type M. tuberculosis H37Rv (Rv) or M. tuberculosis H37Rv Δ-mce1 (Yk). Samples from the ‘early’ time-points post H37Rv-infection (15 min 30 min and 60 min) are colored yellow/orange, and samples from the ‘late’ time-points post H37Rv-infection (4 hrs and 10 hrs) are colored red/brown. Samples from the ‘early’ time-points post Δ-mce1-H37Rv infection (15 min, 30 min, and 60 min) are colored light green, and samples from the ‘late’ time-points post Δ-mce1-H37Rv -infection (4 hrs and 10 hrs]) are dark green/black. The uninfected samples (U) are colored purple, and the pooled common reference samples are colored blue.

    Article Snippet: Eight-hundred-twenty-five ng of Cy-5 labeled cRNA from M. tuberculosis H37Rv or Δ-mce1 H37Rv infected J774A.1 MΦ was randomly hybridized against 825 ng Cy-3 labeled pooled common reference cRNA onto 4×44K 60-mer oligo whole mouse genome - micro-arrays (Agilent Technologies, Santa Clara, USA) using the Agilent Gene Expression hybridization kit (# 5188–5242) as described in the Two-Color Microarray-Based Gene Expression Analysis v5.5 manual (Agilent Technologies, Santa Clara, USA).

    Techniques: Infection

    Transcriptional profile of the genes selected for RT-QPCR verification. Each data point represents the relative quantification of gene expression, by J774A.1 murine macrophages, between the pooled common reference and the different time-points post M. tuberculosis H37Rv (Rv) or M. tuberculosis Δ- mce1 H37Rv (Yk) infection.

    Journal: PLoS ONE

    Article Title: Modulation of Transcriptional and Inflammatory Responses in Murine Macrophages by the Mycobacterium tuberculosis Mammalian Cell Entry (Mce) 1 Complex

    doi: 10.1371/journal.pone.0026295

    Figure Lengend Snippet: Transcriptional profile of the genes selected for RT-QPCR verification. Each data point represents the relative quantification of gene expression, by J774A.1 murine macrophages, between the pooled common reference and the different time-points post M. tuberculosis H37Rv (Rv) or M. tuberculosis Δ- mce1 H37Rv (Yk) infection.

    Article Snippet: Eight-hundred-twenty-five ng of Cy-5 labeled cRNA from M. tuberculosis H37Rv or Δ-mce1 H37Rv infected J774A.1 MΦ was randomly hybridized against 825 ng Cy-3 labeled pooled common reference cRNA onto 4×44K 60-mer oligo whole mouse genome - micro-arrays (Agilent Technologies, Santa Clara, USA) using the Agilent Gene Expression hybridization kit (# 5188–5242) as described in the Two-Color Microarray-Based Gene Expression Analysis v5.5 manual (Agilent Technologies, Santa Clara, USA).

    Techniques: Quantitative RT-PCR, Expressing, Infection

    Over-represented biological processes among up-regulated (A) and down-regulated (B) genes in J774A.1 murine macrophages following infection with M. tuberculosis H37Rv (Rv) or M. tuberculosis Δ- mce1 H37Rv (Yk). The color intensity indicates the negative log of the p-values, dark values representing highly functional processes significantly over-represented among the genes. The numbers presented on the heat map display the percentage of genes within a gene set that map to a certain term, e.g. 19.7% of the 432 genes up-regulated (A) 15 min post H37Rv infection map to the biological process ‘Response to stimulus’. The first column depicts the overall distribution of a term among the 13, 990 genes with detectable expression in the data set, followed by the gene sets for the 5 time-points post H37Rv-infection and the 5 time-points post Δ- mce1 -H37Rv infection.

    Journal: PLoS ONE

    Article Title: Modulation of Transcriptional and Inflammatory Responses in Murine Macrophages by the Mycobacterium tuberculosis Mammalian Cell Entry (Mce) 1 Complex

    doi: 10.1371/journal.pone.0026295

    Figure Lengend Snippet: Over-represented biological processes among up-regulated (A) and down-regulated (B) genes in J774A.1 murine macrophages following infection with M. tuberculosis H37Rv (Rv) or M. tuberculosis Δ- mce1 H37Rv (Yk). The color intensity indicates the negative log of the p-values, dark values representing highly functional processes significantly over-represented among the genes. The numbers presented on the heat map display the percentage of genes within a gene set that map to a certain term, e.g. 19.7% of the 432 genes up-regulated (A) 15 min post H37Rv infection map to the biological process ‘Response to stimulus’. The first column depicts the overall distribution of a term among the 13, 990 genes with detectable expression in the data set, followed by the gene sets for the 5 time-points post H37Rv-infection and the 5 time-points post Δ- mce1 -H37Rv infection.

    Article Snippet: Eight-hundred-twenty-five ng of Cy-5 labeled cRNA from M. tuberculosis H37Rv or Δ-mce1 H37Rv infected J774A.1 MΦ was randomly hybridized against 825 ng Cy-3 labeled pooled common reference cRNA onto 4×44K 60-mer oligo whole mouse genome - micro-arrays (Agilent Technologies, Santa Clara, USA) using the Agilent Gene Expression hybridization kit (# 5188–5242) as described in the Two-Color Microarray-Based Gene Expression Analysis v5.5 manual (Agilent Technologies, Santa Clara, USA).

    Techniques: Infection, Functional Assay, Expressing

    Cytokine profiles of cell supernatants secreted from M. tuberculosis H37Rv (white bars) or M. tuberculosis Δ- mce1 H37Rv (grey bars) infected J774A.1 murine macrophages. The numbers below each column reflect the time-points post-infection: 1; uninfected, 2; 15 min, 3; 30 min, 4; 60 min, 5; 4 hrs, and 6; 10 hrs.

    Journal: PLoS ONE

    Article Title: Modulation of Transcriptional and Inflammatory Responses in Murine Macrophages by the Mycobacterium tuberculosis Mammalian Cell Entry (Mce) 1 Complex

    doi: 10.1371/journal.pone.0026295

    Figure Lengend Snippet: Cytokine profiles of cell supernatants secreted from M. tuberculosis H37Rv (white bars) or M. tuberculosis Δ- mce1 H37Rv (grey bars) infected J774A.1 murine macrophages. The numbers below each column reflect the time-points post-infection: 1; uninfected, 2; 15 min, 3; 30 min, 4; 60 min, 5; 4 hrs, and 6; 10 hrs.

    Article Snippet: Eight-hundred-twenty-five ng of Cy-5 labeled cRNA from M. tuberculosis H37Rv or Δ-mce1 H37Rv infected J774A.1 MΦ was randomly hybridized against 825 ng Cy-3 labeled pooled common reference cRNA onto 4×44K 60-mer oligo whole mouse genome - micro-arrays (Agilent Technologies, Santa Clara, USA) using the Agilent Gene Expression hybridization kit (# 5188–5242) as described in the Two-Color Microarray-Based Gene Expression Analysis v5.5 manual (Agilent Technologies, Santa Clara, USA).

    Techniques: Infection

    Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis H37Rv intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Discovery of Novel Oral Protein Synthesis Inhibitors of Mycobacterium tuberculosis That Target Leucyl-tRNA Synthetase

    doi: 10.1128/AAC.01339-16

    Figure Lengend Snippet: Efficacy of compound 14 in a mouse model of acute TB infection under different dosing regimes of once a day (QD), twice a day (BID), or every other day (q48h). C57BL/6J mice were infected with M. tuberculosis H37Rv intratracheally (∼10 5 CFU) and were dosed starting on the following day after infection for 8 days. Only one dose was administered on day 8 under the BID schedule. Mice were sacrificed at least 24 h after the last drug administration. Every dot represents one mouse data point except for linezolid (mean of data for 5 mice ± standard deviation).

    Article Snippet: N-terminal six-histidine-tagged LeuRS from M. tuberculosis H37Rv, which was codon optimized for E. coli (GenScript, Piscataway, NJ, USA), was overexpressed and purified according to the manufacturer's instructions (Novagen, Madison, WI, USA), using an E. coli BL21(DE3) T7 RNA polymerase overexpression strain.

    Techniques: Infection, Mouse Assay, Standard Deviation

    M. tuberculosis H37Rv in vitro kill kinetics. Cells were incubated with compounds at 20 times their MIC values for different times over 14 days in 10 ml of Middlebrook 7H9 medium containing 10% (vol/vol) ADC and 0.05% (vol/vol) Tween 80. The MIC values used in this experiment were 0.013 μg/ml, 0.6 μg/ml, and 0.06 μg/ml for compound 14, linezolid, and moxifloxacin, respectively. The means and the standard deviations of data from triplicate cultures at each point are shown.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Discovery of Novel Oral Protein Synthesis Inhibitors of Mycobacterium tuberculosis That Target Leucyl-tRNA Synthetase

    doi: 10.1128/AAC.01339-16

    Figure Lengend Snippet: M. tuberculosis H37Rv in vitro kill kinetics. Cells were incubated with compounds at 20 times their MIC values for different times over 14 days in 10 ml of Middlebrook 7H9 medium containing 10% (vol/vol) ADC and 0.05% (vol/vol) Tween 80. The MIC values used in this experiment were 0.013 μg/ml, 0.6 μg/ml, and 0.06 μg/ml for compound 14, linezolid, and moxifloxacin, respectively. The means and the standard deviations of data from triplicate cultures at each point are shown.

    Article Snippet: N-terminal six-histidine-tagged LeuRS from M. tuberculosis H37Rv, which was codon optimized for E. coli (GenScript, Piscataway, NJ, USA), was overexpressed and purified according to the manufacturer's instructions (Novagen, Madison, WI, USA), using an E. coli BL21(DE3) T7 RNA polymerase overexpression strain.

    Techniques: In Vitro, Incubation

    Efficacy of compound 14 in a mouse model of chronic TB infection. C57BL/6J mice were infected with M. tuberculosis H37Rv intratracheally (∼10 2 CFU) and were dosed once daily for 8 weeks starting 6 weeks after infection. Mice were sacrificed 24 h after the last drug administration. Every column represents the mean values ± standard deviations of data from 7 mice per group for untreated and linezolid-treated groups and from 3 mice for compound 14-treated mice. *, P

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Discovery of Novel Oral Protein Synthesis Inhibitors of Mycobacterium tuberculosis That Target Leucyl-tRNA Synthetase

    doi: 10.1128/AAC.01339-16

    Figure Lengend Snippet: Efficacy of compound 14 in a mouse model of chronic TB infection. C57BL/6J mice were infected with M. tuberculosis H37Rv intratracheally (∼10 2 CFU) and were dosed once daily for 8 weeks starting 6 weeks after infection. Mice were sacrificed 24 h after the last drug administration. Every column represents the mean values ± standard deviations of data from 7 mice per group for untreated and linezolid-treated groups and from 3 mice for compound 14-treated mice. *, P

    Article Snippet: N-terminal six-histidine-tagged LeuRS from M. tuberculosis H37Rv, which was codon optimized for E. coli (GenScript, Piscataway, NJ, USA), was overexpressed and purified according to the manufacturer's instructions (Novagen, Madison, WI, USA), using an E. coli BL21(DE3) T7 RNA polymerase overexpression strain.

    Techniques: Infection, Mouse Assay

    Deletion of nuoG in BCG improves protection of mice against TB. (A) Schematic design of protection studies. (B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), or BCG Δ nuoG (▼) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain H37Rv. Bacterial burdens of lungs and spleen were determined at 30 and 180 days postchallenge by plating organ homogenates on agar. Shown are means ± standard deviations. Results are representative of two experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test ( n = 8). *, P

    Journal: mBio

    Article Title: Deletion of nuoG from the Vaccine Candidate Mycobacterium bovis BCG ΔureC::hly Improves Protection against Tuberculosis

    doi: 10.1128/mBio.00679-16

    Figure Lengend Snippet: Deletion of nuoG in BCG improves protection of mice against TB. (A) Schematic design of protection studies. (B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), or BCG Δ nuoG (▼) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain H37Rv. Bacterial burdens of lungs and spleen were determined at 30 and 180 days postchallenge by plating organ homogenates on agar. Shown are means ± standard deviations. Results are representative of two experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test ( n = 8). *, P

    Article Snippet: Two experiments; n = 7 to 8. (G) Frequencies of IFN-γ+ CD4+ T cells were measured in lymph nodes by intracellular cytokine staining following restimulation with M. tuberculosis H37Rv lysate.

    Techniques: Mouse Assay, Infection

    Deletion of nuoG in the clinical vaccine candidate, BCG Δ ureC :: hly , further improves long-term protection against TB. (A and B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), BCG Δ ureC :: hly (▲), or BCG Δ ureC :: hly Δ nuoG (◆) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain H37Rv ( n = 5 to 6) per animal (A) or a clinical isolate of the Beijing/W lineage ( n = 7) (B). Bacterial burdens of organs were assessed at designated time points p.i. by plating organ homogenates. Shown are means ± standard deviations. Results are representative of three (A) or two (B) experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test. (C) Gross pathology of left lung lobe from three out of six vaccinated mice 180 days p.i. with M. tuberculosis H37Rv. Bar, 5 mm. Results are representative of two experiments. (D) Pulmonary histopathology of the left lung lobe from vaccinated mice 180 days p.i. with M. tuberculosis H37Rv. Lungs were fixed and embedded, and sections were stained with H E prior to examination. Bar, 1 mm. Results are representative of two experiments. (E) Survival of SCID mice after subcutaneous administration of 10 6 CFU of indicated strains was monitored over time. Median survival calculated by Mantel-Cox log rank test was 181.5 days (BCG SSI), 398 days (BCG Δ ureC :: hly ), and 405 days (BCG Δ ureC :: hly Δ nuoG ). Results are representative of two experiments. *, P

    Journal: mBio

    Article Title: Deletion of nuoG from the Vaccine Candidate Mycobacterium bovis BCG ΔureC::hly Improves Protection against Tuberculosis

    doi: 10.1128/mBio.00679-16

    Figure Lengend Snippet: Deletion of nuoG in the clinical vaccine candidate, BCG Δ ureC :: hly , further improves long-term protection against TB. (A and B) Mice vaccinated subcutaneously with phosphate-buffered saline (PBS; ●), BCG SSI (■), BCG Δ ureC :: hly (▲), or BCG Δ ureC :: hly Δ nuoG (◆) were aerosol infected after 90 days with a low dose of 100 to 200 CFU of M. tuberculosis laboratory strain H37Rv ( n = 5 to 6) per animal (A) or a clinical isolate of the Beijing/W lineage ( n = 7) (B). Bacterial burdens of organs were assessed at designated time points p.i. by plating organ homogenates. Shown are means ± standard deviations. Results are representative of three (A) or two (B) experiments. Data were analyzed using one-way ANOVA with Tukey’s multiple-comparison test. (C) Gross pathology of left lung lobe from three out of six vaccinated mice 180 days p.i. with M. tuberculosis H37Rv. Bar, 5 mm. Results are representative of two experiments. (D) Pulmonary histopathology of the left lung lobe from vaccinated mice 180 days p.i. with M. tuberculosis H37Rv. Lungs were fixed and embedded, and sections were stained with H E prior to examination. Bar, 1 mm. Results are representative of two experiments. (E) Survival of SCID mice after subcutaneous administration of 10 6 CFU of indicated strains was monitored over time. Median survival calculated by Mantel-Cox log rank test was 181.5 days (BCG SSI), 398 days (BCG Δ ureC :: hly ), and 405 days (BCG Δ ureC :: hly Δ nuoG ). Results are representative of two experiments. *, P

    Article Snippet: Two experiments; n = 7 to 8. (G) Frequencies of IFN-γ+ CD4+ T cells were measured in lymph nodes by intracellular cytokine staining following restimulation with M. tuberculosis H37Rv lysate.

    Techniques: Mouse Assay, Infection, Histopathology, Staining

    CD40 ligand (CD40L) induced antimicrobial activity against Mycobacterium tuberculosis . Primary human monocytes were infected with M. tuberculosis H37Rv and cultured with medium, soluble CD40 L (sCD40L) and/or recombinant IFN-γ (rIFN-γ)

    Journal: Immunology

    Article Title: CD40 ligand and interferon-? induce an antimicrobial response against Mycobacterium tuberculosis in human monocytes

    doi: 10.1111/imm.12062

    Figure Lengend Snippet: CD40 ligand (CD40L) induced antimicrobial activity against Mycobacterium tuberculosis . Primary human monocytes were infected with M. tuberculosis H37Rv and cultured with medium, soluble CD40 L (sCD40L) and/or recombinant IFN-γ (rIFN-γ)

    Article Snippet: The M. tuberculosis H37Rv Whole Cell Lysate ( M. tuberculosis sonicate) was from BEI Resources (Manassas, VA).

    Techniques: Activity Assay, Infection, Cell Culture, Recombinant

    TAC-HRM of rrs (amplicon 3) and gyrB (amplicon 2) gene segments. Singleplex PCR mixtures with primers and probes were loaded into empty microfluidic cards (1 sample per port, yielding 48 reaction mixtures per sample). Results are shown for probe-based detection (A and E), for amplification with SYTO9 (B and F), and for melt curve and difference curve analysis (C and D, G and H). The aligned melt curves show melt temperature shifts versus those for the M. tuberculosis H37Rv wild-type control based on nucleotide changes (leftward variation indicates a lower melt temperature and rightward variation a higher melt temperature). The difference plot uses the same data but plots the negative first derivative (− dF / dt ) on the y axis. The HRM software automatically labels samples that are variants from the wild-type control with a different color. Underlining indicates nucleotides that changed. The rrs (amplicon 3) primer/probe results show the amplification curve obtained with the rrs -A(1401)G-specific probe for a mutant isolate, while the results for M. tuberculosis H37Rv were negative. Both the mutant isolate and H37Rv amplified with the primers using SYTO9, and the HRM result reinforced the probe result as being a mutant/variant sample. The gyrB (amplicon 2) primer/probe results demonstrated the benefit of combining the probe with HRM, since the specific probe (E) detected the Asp461His (GAC→CAC) transversion, while HRM would not (F, G, and H); however, HRM detected the rare mutations Asp461Ala and Asp461Asn.

    Journal: mBio

    Article Title: Integrated Microfluidic Card with TaqMan Probes and High-Resolution Melt Analysis To Detect Tuberculosis Drug Resistance Mutations across 10 Genes

    doi: 10.1128/mBio.02273-14

    Figure Lengend Snippet: TAC-HRM of rrs (amplicon 3) and gyrB (amplicon 2) gene segments. Singleplex PCR mixtures with primers and probes were loaded into empty microfluidic cards (1 sample per port, yielding 48 reaction mixtures per sample). Results are shown for probe-based detection (A and E), for amplification with SYTO9 (B and F), and for melt curve and difference curve analysis (C and D, G and H). The aligned melt curves show melt temperature shifts versus those for the M. tuberculosis H37Rv wild-type control based on nucleotide changes (leftward variation indicates a lower melt temperature and rightward variation a higher melt temperature). The difference plot uses the same data but plots the negative first derivative (− dF / dt ) on the y axis. The HRM software automatically labels samples that are variants from the wild-type control with a different color. Underlining indicates nucleotides that changed. The rrs (amplicon 3) primer/probe results show the amplification curve obtained with the rrs -A(1401)G-specific probe for a mutant isolate, while the results for M. tuberculosis H37Rv were negative. Both the mutant isolate and H37Rv amplified with the primers using SYTO9, and the HRM result reinforced the probe result as being a mutant/variant sample. The gyrB (amplicon 2) primer/probe results demonstrated the benefit of combining the probe with HRM, since the specific probe (E) detected the Asp461His (GAC→CAC) transversion, while HRM would not (F, G, and H); however, HRM detected the rare mutations Asp461Ala and Asp461Asn.

    Article Snippet: The mycobacterial strains used in this study included M. tuberculosis H37Rv (ATCC 27294) and a total of 230 clinical isolates, including 92 from Bangladesh (International Centre for Diarrheal Diseases and Research, Dhaka), 85 from Thailand (Department of Microbiology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok), 28 from the Russian Federation (Scientific Center of Family Health and Reproductive Problems, Siberian Branch, Russian Academy of Medical Sciences, Irkutsk), 23 from Tanzania (Kilimanjaro Clinical Research Institute, Moshi), and 2 from the University of Virginia.

    Techniques: Amplification, Polymerase Chain Reaction, Software, Mutagenesis, Variant Assay

    Growth containment of virulent M. tuberculosis H37Rv organisms induced in peritoneal macrophages by M. tuberculosis H37Ra-primed lymphocytes. Peritoneal macrophages were infected with live, dispersed virulent M. tuberculosis H37Rv at 37°C for 2 to 4 h and then cocultured for 7 days with different numbers of syngeneic splenocytes derived from M. tuberculosis H37Ra-vaccinated and nonvaccinated inbred strain 2 guinea pigs. The cultures were harvested and streaked onto M7H10 agar plates. After 3 weeks of incubation, the CFU were counted. The means ± standard errors of the means for four animals in each group are shown.

    Journal: Infection and Immunity

    Article Title: Altered Cytokine Production and Impaired Antimycobacterial Immunity in Protein-Malnourished Guinea Pigs

    doi:

    Figure Lengend Snippet: Growth containment of virulent M. tuberculosis H37Rv organisms induced in peritoneal macrophages by M. tuberculosis H37Ra-primed lymphocytes. Peritoneal macrophages were infected with live, dispersed virulent M. tuberculosis H37Rv at 37°C for 2 to 4 h and then cocultured for 7 days with different numbers of syngeneic splenocytes derived from M. tuberculosis H37Ra-vaccinated and nonvaccinated inbred strain 2 guinea pigs. The cultures were harvested and streaked onto M7H10 agar plates. After 3 weeks of incubation, the CFU were counted. The means ± standard errors of the means for four animals in each group are shown.

    Article Snippet: Adherent peritoneal macrophages were cultured with live, dispersed virulent M. tuberculosis H37Rv organisms (ATCC 27294) at a ratio of approximately 100 macrophages per mycobacterium at 37°C for 2 to 4 h. Then differently treated lymphocytes were added, and the cells were cultured for another 7 days.

    Techniques: Infection, Derivative Assay, Incubation

    Influence of dietary protein malnutrition on the interactions between lymphocytes and macrophages. Peritoneal macrophages from normally nourished donors were infected with live, dispersed virulent M. tuberculosis H37Rv at 37°C for 2 to 4 h and then cocultured for 7 days with whole syngeneic splenocytes or splenocytes depleted of CD4 T cells, CD8 T cells, or both. The splenocytes were derived from inbred strain 2 guinea pigs vaccinated with M. tuberculosis H37Ra and maintained on the LP or HP diet. The cultures were harvested and streaked onto M7H10 agar plates. After 3 weeks of incubation, the CFU were counted. The means ± standard errors of the means for four animals in each group are shown.

    Journal: Infection and Immunity

    Article Title: Altered Cytokine Production and Impaired Antimycobacterial Immunity in Protein-Malnourished Guinea Pigs

    doi:

    Figure Lengend Snippet: Influence of dietary protein malnutrition on the interactions between lymphocytes and macrophages. Peritoneal macrophages from normally nourished donors were infected with live, dispersed virulent M. tuberculosis H37Rv at 37°C for 2 to 4 h and then cocultured for 7 days with whole syngeneic splenocytes or splenocytes depleted of CD4 T cells, CD8 T cells, or both. The splenocytes were derived from inbred strain 2 guinea pigs vaccinated with M. tuberculosis H37Ra and maintained on the LP or HP diet. The cultures were harvested and streaked onto M7H10 agar plates. After 3 weeks of incubation, the CFU were counted. The means ± standard errors of the means for four animals in each group are shown.

    Article Snippet: Adherent peritoneal macrophages were cultured with live, dispersed virulent M. tuberculosis H37Rv organisms (ATCC 27294) at a ratio of approximately 100 macrophages per mycobacterium at 37°C for 2 to 4 h. Then differently treated lymphocytes were added, and the cells were cultured for another 7 days.

    Techniques: Infection, Derivative Assay, Incubation

    Kaplan-Meier plot of the survival of guinea pigs after low dose aerosol infection with virulent M. tuberculosis H37Rv. Guinea pigs were inoculated with saline, BCG, CFP+MPL/DDA, Ag85A+MPL/DDA or MPL/DDA and rested for 10 weeks prior to infection. All

    Journal: Tuberculosis (Edinburgh, Scotland)

    Article Title: Assessment of vaccine testing at three laboratories using the guinea pig model of tuberculosis

    doi: 10.1016/j.tube.2011.09.003

    Figure Lengend Snippet: Kaplan-Meier plot of the survival of guinea pigs after low dose aerosol infection with virulent M. tuberculosis H37Rv. Guinea pigs were inoculated with saline, BCG, CFP+MPL/DDA, Ag85A+MPL/DDA or MPL/DDA and rested for 10 weeks prior to infection. All

    Article Snippet: Twelve weeks after the initial immunization, all guinea pigs were challenged by the respiratory route with virulent M. tuberculosis H37Rv (ATCC 25495) using a Madison Aerosol Exposure Chamber according to published protocols., The challenge suspension was generated in a Colison nebulizer (BGI, Inc., USA) from a frozen aliquot (−80°C) prepared as described previously.

    Techniques: Infection

    Vaccination with BCG or BCGΔBCG149c does not change immune cells activation in response to CFP after infection. C57BL/6 mice ( n = 5) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route, and 6 months post-infection, lysates from (A) spleen, (B) lymph nodes, and (C) lungs, and stimulated or not with M. tuberculosis H37Rv CFP. Data (SFUs/2 × 10 5 cells) from individual samples are shown, with the mean value for each group indicated by a horizontal line. Data were analyzed for statistical significance between the groups by ANOVA, and we found variable response from animal to animal, with no statistical significance between vaccinated and non-vaccinated mice.

    Journal: Frontiers in Microbiology

    Article Title: The BCGΔBCG1419c Vaccine Candidate Reduces Lung Pathology, IL-6, TNF-α, and IL-10 During Chronic TB Infection

    doi: 10.3389/fmicb.2018.01281

    Figure Lengend Snippet: Vaccination with BCG or BCGΔBCG149c does not change immune cells activation in response to CFP after infection. C57BL/6 mice ( n = 5) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route, and 6 months post-infection, lysates from (A) spleen, (B) lymph nodes, and (C) lungs, and stimulated or not with M. tuberculosis H37Rv CFP. Data (SFUs/2 × 10 5 cells) from individual samples are shown, with the mean value for each group indicated by a horizontal line. Data were analyzed for statistical significance between the groups by ANOVA, and we found variable response from animal to animal, with no statistical significance between vaccinated and non-vaccinated mice.

    Article Snippet: Cells (2 × 105 ) were then cultured in plates coated with anti-IFN-γ capture antibody according to the manufacturer’s protocol (eBiosciences, San Diego, CA, United States) for 24 h at 37°C, 5% CO2 in RPMI-1640 supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO, United States), 100 U/ml penicillin, 100 μg/ml streptomycin, and 200 mM L -glutamine (Sigma, St. Louis, MO, United States) in the presence of M. tuberculosis H37Rv-derived culture filtrate protein (BEI Resources, Manassas, VA, United States).

    Techniques: Activation Assay, Infection, Mouse Assay

    Vaccination with BCG or BCGΔBCG149c provide protection against M. tuberculosis replication after aerosol infection. C57BL/6 mice ( n = 7) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route and the bacterial load was assessed 6 months later in the (A) lung and (B) spleen. Data (Log 10 CFU) from individual samples are shown, with the mean value indicated by a line. Data were analyzed for statistical significance between the groups by ANOVA, and the resulting p -values are shown.

    Journal: Frontiers in Microbiology

    Article Title: The BCGΔBCG1419c Vaccine Candidate Reduces Lung Pathology, IL-6, TNF-α, and IL-10 During Chronic TB Infection

    doi: 10.3389/fmicb.2018.01281

    Figure Lengend Snippet: Vaccination with BCG or BCGΔBCG149c provide protection against M. tuberculosis replication after aerosol infection. C57BL/6 mice ( n = 7) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route and the bacterial load was assessed 6 months later in the (A) lung and (B) spleen. Data (Log 10 CFU) from individual samples are shown, with the mean value indicated by a line. Data were analyzed for statistical significance between the groups by ANOVA, and the resulting p -values are shown.

    Article Snippet: Cells (2 × 105 ) were then cultured in plates coated with anti-IFN-γ capture antibody according to the manufacturer’s protocol (eBiosciences, San Diego, CA, United States) for 24 h at 37°C, 5% CO2 in RPMI-1640 supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO, United States), 100 U/ml penicillin, 100 μg/ml streptomycin, and 200 mM L -glutamine (Sigma, St. Louis, MO, United States) in the presence of M. tuberculosis H37Rv-derived culture filtrate protein (BEI Resources, Manassas, VA, United States).

    Techniques: Infection, Mouse Assay

    Vaccination with BCGΔBCG149c reduces production of IFN-γ, TNF-α, IL-6, and IL-10 after infection. C57BL/6 mice ( n = 6–7) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route, and 6 months post-infection, lysates from lungs were used to quantify IFN-γ, TNF-α, IL-2, IL-17, IL-6, IL-4, and IL-10. Data (mean pg/ml ± SEM) were analyzed for statistical significance between the groups by ANOVA, and the resulting p -values are shown.

    Journal: Frontiers in Microbiology

    Article Title: The BCGΔBCG1419c Vaccine Candidate Reduces Lung Pathology, IL-6, TNF-α, and IL-10 During Chronic TB Infection

    doi: 10.3389/fmicb.2018.01281

    Figure Lengend Snippet: Vaccination with BCGΔBCG149c reduces production of IFN-γ, TNF-α, IL-6, and IL-10 after infection. C57BL/6 mice ( n = 6–7) were vaccinated with BCG Pasteur or BCGΔBCG149c (s.c. 5 × 10 4 CFU), or mock-vaccinated with sterile saline as control. Four weeks after vaccination, the mice were challenged with approximately 100 CFU of M. tuberculosis H37Rv by aerosol route, and 6 months post-infection, lysates from lungs were used to quantify IFN-γ, TNF-α, IL-2, IL-17, IL-6, IL-4, and IL-10. Data (mean pg/ml ± SEM) were analyzed for statistical significance between the groups by ANOVA, and the resulting p -values are shown.

    Article Snippet: Cells (2 × 105 ) were then cultured in plates coated with anti-IFN-γ capture antibody according to the manufacturer’s protocol (eBiosciences, San Diego, CA, United States) for 24 h at 37°C, 5% CO2 in RPMI-1640 supplemented with 10% fetal bovine serum (FBS; Atlas Biologicals, Fort Collins, CO, United States), 100 U/ml penicillin, 100 μg/ml streptomycin, and 200 mM L -glutamine (Sigma, St. Louis, MO, United States) in the presence of M. tuberculosis H37Rv-derived culture filtrate protein (BEI Resources, Manassas, VA, United States).

    Techniques: Infection, Mouse Assay

    cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis H37Rv-MA or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: cyt- bd confers resistance to PAB compounds. Concentration-response curves for IDR-0341930 against M. tuberculosis (A) or M. smegmatis (B) strains. (C) Kill kinetics of IDR-0341930 against aerobically growing M. tuberculosis H37Rv-MA or H37Rv-MA cydC :: aph . IDR-0341930 was added at 10× the MIC of the individual strains. Data are representative are at least three independent experiments. The dotted line represents the limit of detection (10 bacteria/ml).

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques: Concentration Assay

    PAB has synergistic bactericidal activity in combination with CFZ. IDR-0341930 and CFZ were tested singly and in combination at the indicated multiples of the MIC against H37Rv-LP for 7 days. Liquid cultures from a 96-well plate were spot-plated onto 7H10-OADC agar. Pictures are representative of two independent experiments.

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: PAB has synergistic bactericidal activity in combination with CFZ. IDR-0341930 and CFZ were tested singly and in combination at the indicated multiples of the MIC against H37Rv-LP for 7 days. Liquid cultures from a 96-well plate were spot-plated onto 7H10-OADC agar. Pictures are representative of two independent experiments.

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques: Activity Assay

    PAB-CFZ combination causes enhanced killing of M. tuberculosis under starvation conditions. Kill kinetics under PBS-starved conditions of IDR-0341930 and CFZ at 10× the MIC against H37Rv-LP (A), H37Rv-MA (B), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C). Data are representative of at least two independent experiments. The dotted line represents the lower limit of detection (10 bacteria/ml).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: PAB-CFZ combination causes enhanced killing of M. tuberculosis under starvation conditions. Kill kinetics under PBS-starved conditions of IDR-0341930 and CFZ at 10× the MIC against H37Rv-LP (A), H37Rv-MA (B), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C). Data are representative of at least two independent experiments. The dotted line represents the lower limit of detection (10 bacteria/ml).

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques:

    PAB-CFZ combination causes enhanced killing of M. tuberculosis under replicating conditions. Kill kinetics under replicating conditions of IDR-0341930 at 10× the MIC in combination with CFZ at either 10× the MIC (A to C) or 1× the MIC. Combinations were tested against H37Rv-LP (A and D), H37Rv-MA (B and E), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C and F). Samples were taken at the indicated times. Data are representative of at least two independent experiments. The dotted lines represent the upper and lower limits of detection (10 8 and 10 bacteria/ml, respectively).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: PAB-CFZ combination causes enhanced killing of M. tuberculosis under replicating conditions. Kill kinetics under replicating conditions of IDR-0341930 at 10× the MIC in combination with CFZ at either 10× the MIC (A to C) or 1× the MIC. Combinations were tested against H37Rv-LP (A and D), H37Rv-MA (B and E), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C and F). Samples were taken at the indicated times. Data are representative of at least two independent experiments. The dotted lines represent the upper and lower limits of detection (10 8 and 10 bacteria/ml, respectively).

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques:

    PAB-BDQ combinations are active under starvation conditions but are antagonistic at early time points. Kill kinetics under PBS-starved conditions of IDR-0341930 and BDQ at 10× the MIC against H37Rv-LP (A), H37Rv-MA (B), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C). Data are representative of at least two independent experiments. The dotted line represents the lower limit of detection (10 bacteria/ml).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: PAB-BDQ combinations are active under starvation conditions but are antagonistic at early time points. Kill kinetics under PBS-starved conditions of IDR-0341930 and BDQ at 10× the MIC against H37Rv-LP (A), H37Rv-MA (B), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C). Data are representative of at least two independent experiments. The dotted line represents the lower limit of detection (10 bacteria/ml).

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques:

    PAB and BDQ show minimal synergistic activity. Kill kinetics under replicating conditions of IDR-0341930 at 10× the MIC in combination with BDQ at either 10× the MIC (A to C) or 1× the MIC. Combinations were tested against H37Rv-LP (A and D), H37Rv-MA (B and E), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C and F). Samples were taken at the indicated times. Data are representative of at least two independent experiments. The dotted lines represent the upper and lower limits of detection (10 8 and 10 bacteria/ml, respectively).

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Combinations of Respiratory Chain Inhibitors Have Enhanced Bactericidal Activity against Mycobacterium tuberculosis

    doi: 10.1128/AAC.01677-17

    Figure Lengend Snippet: PAB and BDQ show minimal synergistic activity. Kill kinetics under replicating conditions of IDR-0341930 at 10× the MIC in combination with BDQ at either 10× the MIC (A to C) or 1× the MIC. Combinations were tested against H37Rv-LP (A and D), H37Rv-MA (B and E), or H37Rv-MA cydC :: aph (H37Rv-MA cydKO) (C and F). Samples were taken at the indicated times. Data are representative of at least two independent experiments. The dotted lines represent the upper and lower limits of detection (10 8 and 10 bacteria/ml, respectively).

    Article Snippet: M. tuberculosis H37Rv-LP (London Pride) (ATCC 25618) , H37Rv-MA (ATCC 27294) (provided by Chris Sassetti), H37Rv-MA cydC ::aph ( ) (provided by Helena Boshoff), M. smegmatis mc2 155, and M. smegmatis mc2 155 ΔcydA ( ) (provided by Bavesh Kana) were used in this study.

    Techniques: Activity Assay

    Aerosol infection with wild type M. tuberculosis H37Rv (■), mce1R mutant (●), and complemented mce1R mutant (▲). Infected BALB/c mice were given antibiotics for 4 weeks in each group after 4 weeks of infection. The cfu recovery

    Journal: The Journal of infectious diseases

    Article Title: Post-treatment reactivation tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R

    doi: 10.1086/655224

    Figure Lengend Snippet: Aerosol infection with wild type M. tuberculosis H37Rv (■), mce1R mutant (●), and complemented mce1R mutant (▲). Infected BALB/c mice were given antibiotics for 4 weeks in each group after 4 weeks of infection. The cfu recovery

    Article Snippet: M. tuberculosis H37Rv wild type (ATCC 25618), M. tuberculosis H37Rv mce1 operon mutant, M. tuberculosis H37Rv mce1R mutant, complemented M. tuberculosis H37Rv mce1R mutant, and Erdman wild type were grown in Middlebrook 7H9 broth supplemented with 10% ADC (albumin-dextrose-catalase, Becton Dickinson), 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H11 agar containing OADC (oleic acid–albumin-dextrose-catalase supplement, Becton Dickinson), 0.5% glycerol and the antifungal agent cycloheximide (100 mg/ml) (Sigma-Aldrich).

    Techniques: Infection, Mutagenesis, Mouse Assay

    Modified Cornell model of infection with wild type M. tuberculosis H37Rv (■), mce1R mutant (●), and complemented mce1R mutant (▲). BALB/c mice were infected via the inhalation route and antibiotics were given for 8 weeks in each

    Journal: The Journal of infectious diseases

    Article Title: Post-treatment reactivation tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R

    doi: 10.1086/655224

    Figure Lengend Snippet: Modified Cornell model of infection with wild type M. tuberculosis H37Rv (■), mce1R mutant (●), and complemented mce1R mutant (▲). BALB/c mice were infected via the inhalation route and antibiotics were given for 8 weeks in each

    Article Snippet: M. tuberculosis H37Rv wild type (ATCC 25618), M. tuberculosis H37Rv mce1 operon mutant, M. tuberculosis H37Rv mce1R mutant, complemented M. tuberculosis H37Rv mce1R mutant, and Erdman wild type were grown in Middlebrook 7H9 broth supplemented with 10% ADC (albumin-dextrose-catalase, Becton Dickinson), 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H11 agar containing OADC (oleic acid–albumin-dextrose-catalase supplement, Becton Dickinson), 0.5% glycerol and the antifungal agent cycloheximide (100 mg/ml) (Sigma-Aldrich).

    Techniques: Modification, Infection, Mutagenesis, Mouse Assay

    Tail-vein infection with low-dose (5 × 10 2 ) Erdman wild type (◆), M. tuberculosis H37Rv wild type (■), mce1 operon mutant (▲), and mce1R mutant (●). Antibiotics were given for 4 weeks in each group. Mice in panel

    Journal: The Journal of infectious diseases

    Article Title: Post-treatment reactivation tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R

    doi: 10.1086/655224

    Figure Lengend Snippet: Tail-vein infection with low-dose (5 × 10 2 ) Erdman wild type (◆), M. tuberculosis H37Rv wild type (■), mce1 operon mutant (▲), and mce1R mutant (●). Antibiotics were given for 4 weeks in each group. Mice in panel

    Article Snippet: M. tuberculosis H37Rv wild type (ATCC 25618), M. tuberculosis H37Rv mce1 operon mutant, M. tuberculosis H37Rv mce1R mutant, complemented M. tuberculosis H37Rv mce1R mutant, and Erdman wild type were grown in Middlebrook 7H9 broth supplemented with 10% ADC (albumin-dextrose-catalase, Becton Dickinson), 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H11 agar containing OADC (oleic acid–albumin-dextrose-catalase supplement, Becton Dickinson), 0.5% glycerol and the antifungal agent cycloheximide (100 mg/ml) (Sigma-Aldrich).

    Techniques: Infection, Mutagenesis, Mouse Assay

    Histology (H E stain) of lung sections of mice infected for 4 weeks before treatment was initiated (row 1), at the end of 8 weeks of treatment (row 2), and 20 weeks after end of an 8-week treatment (row 3). Mice were infected with H37Rv (column

    Journal: The Journal of infectious diseases

    Article Title: Post-treatment reactivation tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R

    doi: 10.1086/655224

    Figure Lengend Snippet: Histology (H E stain) of lung sections of mice infected for 4 weeks before treatment was initiated (row 1), at the end of 8 weeks of treatment (row 2), and 20 weeks after end of an 8-week treatment (row 3). Mice were infected with H37Rv (column

    Article Snippet: M. tuberculosis H37Rv wild type (ATCC 25618), M. tuberculosis H37Rv mce1 operon mutant, M. tuberculosis H37Rv mce1R mutant, complemented M. tuberculosis H37Rv mce1R mutant, and Erdman wild type were grown in Middlebrook 7H9 broth supplemented with 10% ADC (albumin-dextrose-catalase, Becton Dickinson), 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H11 agar containing OADC (oleic acid–albumin-dextrose-catalase supplement, Becton Dickinson), 0.5% glycerol and the antifungal agent cycloheximide (100 mg/ml) (Sigma-Aldrich).

    Techniques: H&E Stain, Mouse Assay, Infection

    Histopathology of lung sections from BALB/c mice infected with wild type M. tuberculosis H37Rv (A), complemented mce1R mutant (B), and mce1R mutant (C). The mice were infected via the inhalation route and medicated for 4 weeks as described in .

    Journal: The Journal of infectious diseases

    Article Title: Post-treatment reactivation tuberculosis in mice caused by Mycobacterium tuberculosis disrupted in mce1R

    doi: 10.1086/655224

    Figure Lengend Snippet: Histopathology of lung sections from BALB/c mice infected with wild type M. tuberculosis H37Rv (A), complemented mce1R mutant (B), and mce1R mutant (C). The mice were infected via the inhalation route and medicated for 4 weeks as described in .

    Article Snippet: M. tuberculosis H37Rv wild type (ATCC 25618), M. tuberculosis H37Rv mce1 operon mutant, M. tuberculosis H37Rv mce1R mutant, complemented M. tuberculosis H37Rv mce1R mutant, and Erdman wild type were grown in Middlebrook 7H9 broth supplemented with 10% ADC (albumin-dextrose-catalase, Becton Dickinson), 0.2% glycerol and 0.05% Tween 80 or on Middlebrook 7H11 agar containing OADC (oleic acid–albumin-dextrose-catalase supplement, Becton Dickinson), 0.5% glycerol and the antifungal agent cycloheximide (100 mg/ml) (Sigma-Aldrich).

    Techniques: Histopathology, Mouse Assay, Infection, Mutagenesis

    PCR amplification of leuA genes from M. tuberculosis strains . leuA genes were PCR amplified from H37Rv and Amnatchareon strain 731 with two and 14 copies of the tandem repeats, respectively. Lane M, 200 bp DNA markers.

    Journal: BMC Microbiology

    Article Title: Characterization of ?-isopropylmalate synthases containing different copy numbers of tandem repeats in Mycobacterium tuberculosis

    doi: 10.1186/1471-2180-9-122

    Figure Lengend Snippet: PCR amplification of leuA genes from M. tuberculosis strains . leuA genes were PCR amplified from H37Rv and Amnatchareon strain 731 with two and 14 copies of the tandem repeats, respectively. Lane M, 200 bp DNA markers.

    Article Snippet: M. tuberculosis isolate number 731, obtained from a pulmonary tuberculosis patient in Amnatchareon province, Thailand, and the M. tuberculosis H37Rv strain (laboratory strain: ATCC 25618) were the sources of the leuA gene with 14 and 2 copies, respectively, of the 57 bp tandem repeat [ ].

    Techniques: Polymerase Chain Reaction, Amplification

    Construction of conditional mmpL3 knockdown strains of M. tuberculosis . Allelic replacement at the mmpL3 locus of M. tuberculosis H37Rv strain ATCC 25618 rescued with mmpL3-DAS + 4 expressed from pGMCH-T38S38-P750-mmpL3-DAS + 4 under the control of an

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

    doi: 10.1128/AAC.00826-16

    Figure Lengend Snippet: Construction of conditional mmpL3 knockdown strains of M. tuberculosis . Allelic replacement at the mmpL3 locus of M. tuberculosis H37Rv strain ATCC 25618 rescued with mmpL3-DAS + 4 expressed from pGMCH-T38S38-P750-mmpL3-DAS + 4 under the control of an

    Article Snippet: M. tuberculosis H37Rv ATCC 25618 was grown in Middlebrook 7H9 broth supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD, Difco) and 0.05% Tween 80 or on solid Middlebrook 7H11 medium supplemented with OADC at 37°C.

    Techniques:

    MmpL3 is required for the replication and persistence of M. tuberculosis in mice. (A) Growth and survival and WT M. tuberculosis H37Rv ATCC 25618 (blue line and circles) and MmpL3-DUC (triangles) in C57BL/6 mouse lungs (left) and spleens (right). Mice

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

    doi: 10.1128/AAC.00826-16

    Figure Lengend Snippet: MmpL3 is required for the replication and persistence of M. tuberculosis in mice. (A) Growth and survival and WT M. tuberculosis H37Rv ATCC 25618 (blue line and circles) and MmpL3-DUC (triangles) in C57BL/6 mouse lungs (left) and spleens (right). Mice

    Article Snippet: M. tuberculosis H37Rv ATCC 25618 was grown in Middlebrook 7H9 broth supplemented with oleic acid-albumin-dextrose-catalase (OADC; BD, Difco) and 0.05% Tween 80 or on solid Middlebrook 7H11 medium supplemented with OADC at 37°C.

    Techniques: Mouse Assay

    Effect of depleting MmpL3 on the susceptibility of M. tuberculosis to MmpL3 inhibitors. The zones of inhibition on 7H11-OADC agar plates for each drug were measured versus MmpL3-DUC and the M. tuberculosis H37Rv parent strain in the presence and absence

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

    doi: 10.1128/AAC.00826-16

    Figure Lengend Snippet: Effect of depleting MmpL3 on the susceptibility of M. tuberculosis to MmpL3 inhibitors. The zones of inhibition on 7H11-OADC agar plates for each drug were measured versus MmpL3-DUC and the M. tuberculosis H37Rv parent strain in the presence and absence

    Article Snippet: Six- to 8-week-old female C57BL/6 mice (Jackson Laboratories) were infected with the wild-type and virulent M. tuberculosis H37Rv ATCC 25618 MmpL3-DUC knockdown using established protocols for low-dose aerosol infection ( ) to deliver ∼100 bacilli per mouse.

    Techniques: Inhibition

    Effect of depleting MmpL3 on the viability of nonreplicating hypoxic M. tuberculosis bacilli. Shown is survival of M. tuberculosis ATCC 25618 MmpL3-DUC during hypoxia upon addition of ATc (100 ng/ml) to the culture medium 9 and 15 days after the culture

    Journal: Antimicrobial Agents and Chemotherapy

    Article Title: Therapeutic Potential of the Mycobacterium tuberculosis Mycolic Acid Transporter, MmpL3

    doi: 10.1128/AAC.00826-16

    Figure Lengend Snippet: Effect of depleting MmpL3 on the viability of nonreplicating hypoxic M. tuberculosis bacilli. Shown is survival of M. tuberculosis ATCC 25618 MmpL3-DUC during hypoxia upon addition of ATc (100 ng/ml) to the culture medium 9 and 15 days after the culture

    Article Snippet: Six- to 8-week-old female C57BL/6 mice (Jackson Laboratories) were infected with the wild-type and virulent M. tuberculosis H37Rv ATCC 25618 MmpL3-DUC knockdown using established protocols for low-dose aerosol infection ( ) to deliver ∼100 bacilli per mouse.

    Techniques: