Journal: The Lancet. Microbe

Article Title: Signatures of transmission in within-host Mycobacterium tuberculosis complex variation: a retrospective genomic epidemiology study

doi: 10.1016/j.lanmic.2024.06.003

Figure Lengend Snippet: Characteristics of Mycobacterium tuberculosis complex household transmission studies

Article Snippet: Culture , Isolates cultured on Lowenstein–Jensen slants; each strain plated on Middlebrook 7H10 agar; three loops of culture were scraped and suspended in buffer , Isolates revived from frozen archival stocks on Lowenstein–Jensen slants or in MGIT liquid medium , Cultures obtained from frozen archival stocks; all cultures were grown in MGIT containing modified Middlebrooks 7H9 liquid medium and on Lowenstein–Jensen agar.

Techniques: Transmission Assay, Biomarker Discovery, Cell Culture, Modification, DNA Extraction, Isolation, Disruption, Extraction, Purification, Sequencing

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Mycobacterium tuberculosis PE31 ( Rv3477 ) Attenuates Host Cell Apoptosis and Promotes Recombinant M. smegmatis Intracellular Survival via Up-regulating GTPase Guanylate Binding Protein-1

doi: 10.3389/fcimb.2020.00040

Figure Lengend Snippet: Ms_PE31 can alter the surface characteristics of M. smegmatis . (A) PE31 (Rv3477) sequence amplified by PCR, applying M. tuberculosis genome, ~297 bp in length (lane 1 = DNA ladder and lane 2 = amplified gene). (B) The Ace-induced recombinant cell lysates were prepared and employed the Western blot for confirmation of His-tagged PE31 protein expression. (C) Both recombinant strains cultured on plates containing 7H9 solid media with hyg and Ace (0.02%, w/v), the developed single colony (D) Recombinant strains cultured in Ace (0.02%, w/v) added 7H9 liquid on polystyrene plates to induce the development of biofilm. (E) The quantification of biofilm formation was confirmed by Tetrahydrofuran (THF) assay ( n = 3). Results were determined by Student's t -test, * P < 0.05. Error bars represent the standard deviation of mean.

Article Snippet: Ms_PE31 and Ms_vec, cultured in 100 μg/ml Hyg supplemented Middlebrook 7H9 liquid medium.

Techniques: Sequencing, Amplification, Recombinant, Western Blot, Expressing, Cell Culture, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Mycobacterium tuberculosis PE31 ( Rv3477 ) Attenuates Host Cell Apoptosis and Promotes Recombinant M. smegmatis Intracellular Survival via Up-regulating GTPase Guanylate Binding Protein-1

doi: 10.3389/fcimb.2020.00040

Figure Lengend Snippet: Ms_PE31 increase growth under multiple stress. (A) The growth rate of recombinant strains was measured under in-vitro low pH condition. Cultured bacteria were harvested and re-cultured in 7H9 (pH = 5) to maintained the OD 600 =0.8. Then, incubated in 37°C and 100 μl taken from it after different time intervals for workable enumeration ( n = 3). (B) The reactive nitrogen stress was examined by spot test, the supplementation of 2 and 5 mM diamide to the 7H9 solid medium to grown the Ms_Vec and Ms_PE31 strains. Growth of bacteria on 7H9 solid media which contained mentioned concentrations of diamide ( n = 3). (C,D) Ms_Ves and Ms_PE31 survival upon exposure to H2O2 and SDS, respectively, examined by disk diffusion technique. Whatman disks used to mottled the different concentrations of H2O2 (10 μl) and SDS (10 μl) ( n = 3). Area of the zone of inhibition was calculated after 3−4 days incubation at 37°C. The results were determined by Student's t -test, * P < 0.05 and ** P < 0.01. Error bars represent the standard deviation of mean.

Article Snippet: Ms_PE31 and Ms_vec, cultured in 100 μg/ml Hyg supplemented Middlebrook 7H9 liquid medium.

Techniques: Recombinant, In Vitro, Cell Culture, Bacteria, Incubation, Spot Test, Diffusion-based Assay, Inhibition, Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Mycobacterium tuberculosis PE31 ( Rv3477 ) Attenuates Host Cell Apoptosis and Promotes Recombinant M. smegmatis Intracellular Survival via Up-regulating GTPase Guanylate Binding Protein-1

doi: 10.3389/fcimb.2020.00040

Figure Lengend Snippet: Survival rate of M. smegmatis recombinant strains. (A) Recombinant strains infected macrophages were laved and lysed SDS (0.025%, w/v) at indicated intervals. 10-fold serial diluted lysed cells were mottled on hyg containing 7H9 solid medium plates. After 3–4 days, CFU was computed ( n = 3). (B) Growth kinetics of recombinant strains were determined by the growth of bacteria in 7H9 liquid added Ace (1%, w/v), Tw (0.05%, v/v) and hyg (100 μg/ml) ( n = 3). Results were determined by Student's t -test, *** P < 0.001. Error bars represent the standard deviation of mean.

Article Snippet: Ms_PE31 and Ms_vec, cultured in 100 μg/ml Hyg supplemented Middlebrook 7H9 liquid medium.

Techniques: Recombinant, Infection, Bacteria, Standard Deviation