Journal: Biochimica et biophysica acta. General subjects

Article Title: A new bioinspired peptide on defensin from C. annuum fruits: Antimicrobial activity, mechanisms of action and therapeutical potential.

doi: 10.1016/j.bbagen.2022.130218

Figure Lengend Snippet: Fig. 9. Effect of CaDef2.1G27-K44 on M. tuberculosis growth in bacterial culture. Bacterial suspensions (1 × 106 CFU well−1) of M. tuberculosis strain H37Rv (A) and clinical isolate M299 (B) were treated or untreated with CaDef2.1G27-K44 (200, 100, 50 and 25 μM) for 5 days. Bacterial growth in the resulted cultures was quantified by MTT test. Data are presented as a percentage of bacterial growth of treated culture compared to the growth of corresponding untreated culture (100%). Negative Control (C−) - Middlebrook 7H9 Broth supplemented with ADC; Positive Control (C+) untreated bacterial suspensions. (C) Mini mum inhibitory concentrations of CaDef2.1G27-K44 suppressing growth of the laboratory M. tuberculosis strain H37Rv and clinical M. tuberculosis isolate M299. The results presented are mean values obtained over three experiments, each done in duplicate. Dose response curves were con structed and * p < 0.05; *** p < 0.001 compared to untreated group (0 μM) determined by Tukey test.

Article Snippet: The laboratory Mycobacterium tuberculosis strain H37Rv (ATCC 27294) and highly virulent M. tuberculosis strain of the Beijing genotype, strain M299, isolated from TB patient in Mozambique and previously evaluated for virulence.

Techniques: Negative Control, Positive 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. 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: International Journal of Molecular Sciences

Article Title: The Next Frontier in Tuberculosis Investigation: Automated Whole Genome Sequencing for Mycobacterium tuberculosis Analysis

doi: 10.3390/ijms25147909

Figure Lengend Snippet: Automated Bacterial WGS Assay Results for MTBs and NTMs.

Article Snippet: ATCC_35822D-2 , Mycobacterium tuberculosis , 4.9 , 4,285,606 , 122 , 60.58.

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: The Next Frontier in Tuberculosis Investigation: Automated Whole Genome Sequencing for Mycobacterium tuberculosis Analysis

doi: 10.3390/ijms25147909

Figure Lengend Snippet: Bacterial Strains Used in This Study.

Article Snippet: ATCC_35822D-2 , Mycobacterium tuberculosis , 4.9 , 4,285,606 , 122 , 60.58.

Techniques: Variant Assay

Journal:

Article Title: Loop-Mediated Isothermal Amplification for Direct Detection of Mycobacterium tuberculosis Complex, M. avium , and M. intracellulare in Sputum Samples

doi: 10.1128/JCM.41.6.2616-2622.2003

Figure Lengend Snippet: Specificity of LAMP primers for identification of mycobacterial species

Article Snippet: Genomic DNAs of M. tuberculosis H37Rv, M. tuberculosis H37Ra, Mycobacterium bovis BCG Tokyo, M. avium ATCC 15769, M. avium ATCC 25291, M. avium ATCC 35718 , M. intracellulare ATCC 13950, M. intracellulare ATCC 35847, and M. intracellulare ATCC 35762 were also extracted from Mycobacterium Growth Indicator Tube (MGIT) broth on the day when positive fluorescence was detected.

Techniques: Amplification

Journal: Frontiers in Microbiology

Article Title: Heterologous prime-boost BCG with DNA vaccine expressing fusion antigens Rv2299c and Ag85A improves protective efficacy against Mycobacterium tuberculosis in mice

doi: 10.3389/fmicb.2022.927031

Figure Lengend Snippet: Preliminary demonstration that DNA vaccines expressing fusion antigens Rv2299c and Ag85A are moderately immunogenic and can afford protection. (A) Vaccination and infection schedule. Six weeks after the primary immunization, the mice were sacrificed, then splenocytes were collected and treated with Rv2299c (10 μg/ml) or Ag85A (10 μg/ml) at 37°C for 15 h in the presence of GolgiStop and intracellular cytokine staining was evaluated by flow cytometry. (B) The overall percentages of CD4 + or CD8 + T cells producing IFN-γ, TNF-α, or IL-2 after stimulation with Rv2299c or Ag85A or 2299c-Ag85A ( n = 3 mice, one-way ANOVA, mean ± SEM). (C) Specific IgG responses to antigen Rv2299c or Ag85A or Rv2299c-Ag85A in serum assayed six weeks after the primary immunization ( n = 3 mice). (D) Pulmonary bacterial burden, Log 10 CFU, at four weeks after Mtb H37Rv challenge. ( n = 5 mice, two tailed P and unpaired t test).Data are representative of two independent experiments with at least three mice per group.* p < 0.05, ** p < 0.01,*** p < 0.001, **** p < 0.0001: a significant difference of treatment groups from the appropriate controls (pVAX1 group). ns , no significant difference.

Article Snippet: To produce the separate recombinant Ag85A and Rv2299c proteins, the corresponding gene was amplified by PCR using Mtb H37Rv ATCC 27294 genomic DNA as a template.

Techniques: Vaccines, Expressing, Infection, Staining, Flow Cytometry, Two Tailed Test

Journal: Frontiers in Microbiology

Article Title: Heterologous prime-boost BCG with DNA vaccine expressing fusion antigens Rv2299c and Ag85A improves protective efficacy against Mycobacterium tuberculosis in mice

doi: 10.3389/fmicb.2022.927031

Figure Lengend Snippet: Enhanced protection by the BCG prime-DNA boost regimen. Six weeks after the primary immunization, mice ( n = 5) were aerosol-challenged with Mtb H37Rv then sacrificed at four weeks post-infection. Bacterial loads in the lungs (A) and spleens (B) are shown as the mean log 10 CFU/organ ± SEM. (C) The histopathology of lung tissues assessed in sections stained by acid-fast staining. Scale bars; 100 μm (top: original magnification ×200) and 50 μm (bottom: original magnification ×400). (D) H&E staining of the lungs of infected mice at four weeks post-infection ( n = 5 mice, one-way ANOVA, mean ± SEM). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001: a significant difference of treatment groups from the appropriate controls (PBS group). ns , no significant difference.

Article Snippet: To produce the separate recombinant Ag85A and Rv2299c proteins, the corresponding gene was amplified by PCR using Mtb H37Rv ATCC 27294 genomic DNA as a template.

Techniques: Aerosol, Infection, Histopathology, Staining

Journal: Frontiers in Microbiology

Article Title: Heterologous prime-boost BCG with DNA vaccine expressing fusion antigens Rv2299c and Ag85A improves protective efficacy against Mycobacterium tuberculosis in mice

doi: 10.3389/fmicb.2022.927031

Figure Lengend Snippet: Polyfunctional CD4 + and CD8 + T-cell populations in the lungs of immunized mice after H37Rv challenge. Four weeks after infection of immunized mice pulmonary lymphocytes were collected and stimulated ex vivo with PPD, Ag85A or Rv2299c or Rv2299c-Ag85A. Frequencies of antigen-specific CD4 + (left) and CD8 + (right) T cells producing single or multiple cytokines were analyzed by flow cytometry. ( n = 5 mice, mean ± SEM).

Article Snippet: To produce the separate recombinant Ag85A and Rv2299c proteins, the corresponding gene was amplified by PCR using Mtb H37Rv ATCC 27294 genomic DNA as a template.

Techniques: Infection, Ex Vivo, Flow Cytometry