Journal: medRxiv

Article Title: Direct detection and quantification of Mycobacterium tuberculosis from clinical samples by high-resolution melt qPCR

doi: 10.64898/2026.03.07.26347851

Figure Lengend Snippet: (A) Amplification plot of H37Rv replicates targeting the RD9 gene. (B) Corresponding melt curve showing a specific melting temperature (Tm) of ∼75°C. H37Rv DNA was serially diluted from 10 6 to 10 1 copies per reaction and analysed in triplicate. (C) Amplification plot obtained after 40 cycles for the dilutions, including the no-template control (NTC). (D) Corresponding melt curve showing a Tm of 73.7 ± 0.12°C. (E) Standard curve generated from the 10-fold serial dilutions, plotting cycle threshold (Ct) versus log copy number per reaction.

Article Snippet: Using the M. tuberculosis H37Rv (ATCC #25618DQ) reference strain, we performed asymmetric PCR with Vent (exo-) DNA polymerase (New England Biolabs).

Techniques: Amplification, Control, Generated

Journal: Cellular microbiology

Article Title: Mycobacterium tuberculosis infection causes different levels of apoptosis and necrosis in human macrophages and alveolar epithelial cells.

doi: 10.1046/j.1462-5822.2003.00312.x

Figure Lengend Snippet: Fig. 1. Comparison of the percentage of apop- tosis and necrosis of U937 macrophages infected with M. tuberculosis H37Rv and H37Ra. The MOI used was 10.

Article Snippet: Bacterial strains and growth media Mycobacterium tuberculosis strains H37Rv and H37Ra were obtained from the American Type Culture Collection (ATCC) and were cultured on Middlebrook 7H10 agar for 20 days.

Techniques: Comparison, Infection

Journal: Cellular microbiology

Article Title: Mycobacterium tuberculosis infection causes different levels of apoptosis and necrosis in human macrophages and alveolar epithelial cells.

doi: 10.1046/j.1462-5822.2003.00312.x

Figure Lengend Snippet: Fig. 3. Alveolar epithelial cell apoptosis after M. tuberculosis H37Rv infection. A549 cells were either uninfected (A and B) or infected (C and D) with M. tuberculosis (MOI 10) and treated with staurosporine as described in Experimental procedures. Microscopic examination of staurosporine- treated cells indicated that: (A) staurosporine-treated uninfected A549 cells underwent nuclear morphological changes typical of apoptosis as demonstrated by the TUNEL assay (B); M. tuberculosis infection before adding staurosporine suppressed staurosporine-induced apoptosis in A549 cells (C and D).

Article Snippet: Bacterial strains and growth media Mycobacterium tuberculosis strains H37Rv and H37Ra were obtained from the American Type Culture Collection (ATCC) and were cultured on Middlebrook 7H10 agar for 20 days.

Techniques: Infection, TUNEL Assay

Journal: Cellular microbiology

Article Title: Mycobacterium tuberculosis infection causes different levels of apoptosis and necrosis in human macrophages and alveolar epithelial cells.

doi: 10.1046/j.1462-5822.2003.00312.x

Figure Lengend Snippet: Fig. 6. A and B. Western blot analysis of bad and bcl-2 proteins in macrophage and alveolar epithelial cells. U937 and A549 cells were infected with M. tuberculosis H37Rv at an MOI 1:10 or cultured with medium alone for 48 h. Concentrated cell lysate-free supernatants were subjected to Western blotting (see Experimental procedures). Line 1, A549 uninfected control; line 2, infected A549; line 3, U937 uninfected control; line 4, infected U937.

Article Snippet: Bacterial strains and growth media Mycobacterium tuberculosis strains H37Rv and H37Ra were obtained from the American Type Culture Collection (ATCC) and were cultured on Middlebrook 7H10 agar for 20 days.

Techniques: Western Blot, Infection, Cell Culture, Control

Journal: Cellular microbiology

Article Title: Mycobacterium tuberculosis infection causes different levels of apoptosis and necrosis in human macrophages and alveolar epithelial cells.

doi: 10.1046/j.1462-5822.2003.00312.x

Figure Lengend Snippet: Fig. 7. Effect of bacterial growth inhibition by rifampicin (protein syn- thesis inhibitor) on apoptosis levels of macrophages (A) and alveolar epithelial cells (B) after M. tuberculosis H37Rv infection. U937 mac- rophages as well as A549 alveolar epithelial cells were infected with M. tuberculosis virulent strain with an MOI of 10. To one part of the cell cultures, rifampicin was added 24 h after infection at a concen- tration 0.1 mg ml-1. Apoptosis response was assessed after 3 and 5 days by Cell Death Detection ELISA. Percentage of apoptosis was calculated in infected; infected/rifampicin-treated macrophages and alveolar epithelial cell populations in comparison with uninfected; uninfected/rifampicin-treated monolayers. *Statistically significant dif- ference between groups (P < 0.05).

Article Snippet: Bacterial strains and growth media Mycobacterium tuberculosis strains H37Rv and H37Ra were obtained from the American Type Culture Collection (ATCC) and were cultured on Middlebrook 7H10 agar for 20 days.

Techniques: Inhibition, Infection, Enzyme-linked Immunosorbent Assay, Comparison

Journal: BMC Biochemistry

Article Title: Sustained activation of mTORC1 in macrophages increases AMPKα-dependent autophagy to maintain cellular homeostasis

doi: 10.1186/s12858-016-0069-6

Figure Lengend Snippet: Mycobacterial infection further increased autophagy in TSC1 KO BMMϕ. a Tsc1 KO macrophages have mTORC1-dependent inhibition of ULK1 at S757. BMMϕ cells from either TSC1 WT or KO BMMϕ were left uninfected or infected with M. tuberculosis strain H37Rv (TB) ( a , b , c ) or BCG ( d , e , f ) at a MOI of 10 for 24 h and lysed for immunoblotting analysis with the indicated antibodies. TSC1 WT and KO BMMϕ were treated with 100 ng/ml rapamycin ( b , d ) or with 100 nM bafilomycin A1 ( e , f ) for 3 h and then lysed for Western blot analysis. c , f Densitometric quantification of LC3-II ( n = 3) was normalized to total Actin in c and e . The arrow indicates LC3B-II. These experiments were repeated three times. BafA1, bafilomycin A1; BCG, Bacillus Calmette - Guérin ; Rapa, rapamycin; TB, M. tuberculosis strain H37Rv, WT, Wild type; KO, knockout; UN, uninfected. *, p < 0.05; **, p <0.01

Article Snippet: Virulent M. tuberculosis H37Rv (ATCC, 25618D-2) was purchased from ATCC.

Techniques: Infection, Inhibition, Western Blot, Knock-Out

Journal: BMC Biochemistry

Article Title: Sustained activation of mTORC1 in macrophages increases AMPKα-dependent autophagy to maintain cellular homeostasis

doi: 10.1186/s12858-016-0069-6

Figure Lengend Snippet: Infection of TSC1 f/f LysM-Cre + mice with M. tuberculosis resulted in lower mycobacterial counts at the time of the arrival of adaptive immune cells. Both TSC1 f/f LysM-Cre + and LysM-Cre - mice were aerogenously infected with M. tuberculosis H37Rv with 200-300 bacilli per lung. Bacterial burdens were measured by CFUs in the lungs ( a ) at 1, 2, 3, and 5 weeks post-aerosol infection and in the spleens ( b ) at 3 and 5 weeks post-aerosol infection ( n = 4 per time point). c Cytokine secretion from lung homogenates was determined by Luminex at 3 weeks post-aerosol infection. The data were shown as mean ± SD is shown ( n = 4). d Flow cytometry analysis of MHCII expression in TSC1 WT and KO macrophages at 3 weeks after M. tuberculosis infection is shown. MFI ± SD is shown ( n = 4). MFI = mean fluorescence intensity. *, p < 0.05

Article Snippet: Virulent M. tuberculosis H37Rv (ATCC, 25618D-2) was purchased from ATCC.

Techniques: Infection, Aerosol, Luminex, Flow Cytometry, Expressing, Fluorescence

Journal: Molecular Systems Biology

Article Title: Path‐seq identifies an essential mycolate remodeling program for mycobacterial host adaptation

doi: 10.15252/msb.20188584

Figure Lengend Snippet: Serial 10‐fold dilutions of MTB H37Rv wild type (wt) and MTB with inducible overexpression of Rv0472c were spotted on 7H10 agar plates with or without ATc. Argentation TLC of 14 C‐labeled methyl esters of mycolic acids (MAMEs) obtained from apolar lipids and delipidated cell wall fractions of MSM wt and MSM with inducible overexpression of MSMEG_ 0916. The α, αʹ, epoxy (e), and cyclopropanated α‐ (X 1 ) MAMEs species are labeled. Faster‐migrating species that co‐migrated with α‐MAMEs and accumulate with induced MSMEG_ 0916 overexpression are indicated as X 2 and X 3 . BCG wt and BCG with inducible overexpression of Rv0472c cultures, labeled with 14 C‐acetate, were induced (+ATc) or uninduced (−ATc) for 4 h or 8 h. The total FAMEs and MAMEs were extracted and analyzed by autoradiography–TLC using equal counts (15,000 cpm) for each lane.

Article Snippet: Mycobacterium tuberculosis H37Rv (kind gift of David Sherman) was also used for Path‐seq experiments.

Techniques: Over Expression, Labeling, Autoradiography

Journal: Molecular Systems Biology

Article Title: Path‐seq identifies an essential mycolate remodeling program for mycobacterial host adaptation

doi: 10.15252/msb.20188584

Figure Lengend Snippet:

Article Snippet: Mycobacterium tuberculosis H37Rv (kind gift of David Sherman) was also used for Path‐seq experiments.

Techniques: Recombinant, Sequencing, Cloning, Protease Inhibitor, Software, Bacteria