Journal: Nucleic Acids Research

Article Title: SigH stress response mediates killing of Mycobacterium tuberculosis by activating nitronaphthofuran prodrugs via induction of Mrx2 expression

doi: 10.1093/nar/gkac1173

Figure Lengend Snippet: nNFs prodrugs are activated by Mrx2 conveying anti-tuberculosis activity. ( A ) In vitro enzymatic activity of Mrx2 WT represented by the time-dependent decrease of nNF-C2 absorbance at 412 nm; ( B ) nitric oxide release in nNF treated M. bovis BCG cultures. NO release in BCG cultures exposed to different concentrations of nNF-C2, nNF-C4, PA-824, and RIF, was quantified in the culture supernatants using the DAN assay; (C, D) phenotypical characterization of mrx2 mutations cloned into pFRA220 and expressed in Δ mrx2 . MIC 90 values (μg/ml) for the nNF derivatives nNF-C2 ( C ) and nNF-C4 ( D ) obtained in M. tuberculosis H37Rv WT (wt), Δ mrx2 , and Δ mrx2 transformants expressing the different mrx2 mutants (Δ mrx2 :: mrx2 MUT ); ( E ) in vitro enzymatic activity of Mrx2 wild type and mutants against compound nNF-C2; ( F ) ribbon representation of the crystal structure of Mrx2 (PDB 4ZIL), showing the localization of the Mrx2 mutations.

Article Snippet: Resazurin (Sigma-Aldrich) was added (i) at day 7 after bacterial inoculation in the case of M. tuberculosis H37Rv ATCC 27294, M. tuberculosis Beijing GC1237 and M. bovis BCG 1173 P2, (ii) at day 3 for M. marinum ATCC BAA-535 and M. avium subsp. hominissuis ATCC 25291, (iii) after 16 hs for M. abscessus ATCC19977 and M. aurum ATCC23366, and (iv) after 4 hs for M. smegmatis mc 2 155 and the rest of non-mycobacteria species.

Techniques: Activity Assay, In Vitro, Clone Assay, Expressing

Journal: Nucleic Acids Research

Article Title: SigH stress response mediates killing of Mycobacterium tuberculosis by activating nitronaphthofuran prodrugs via induction of Mrx2 expression

doi: 10.1093/nar/gkac1173

Figure Lengend Snippet: nNFs prodrugs are activated by Mrx2 conveying anti-tuberculosis activity. ( A ) In vitro enzymatic activity of Mrx2 WT represented by the time-dependent decrease of nNF-C2 absorbance at 412 nm; ( B ) nitric oxide release in nNF treated M. bovis BCG cultures. NO release in BCG cultures exposed to different concentrations of nNF-C2, nNF-C4, PA-824, and RIF, was quantified in the culture supernatants using the DAN assay; (C, D) phenotypical characterization of mrx2 mutations cloned into pFRA220 and expressed in Δ mrx2 . MIC 90 values (μg/ml) for the nNF derivatives nNF-C2 ( C ) and nNF-C4 ( D ) obtained in M. tuberculosis H37Rv WT (wt), Δ mrx2 , and Δ mrx2 transformants expressing the different mrx2 mutants (Δ mrx2 :: mrx2 MUT ); ( E ) in vitro enzymatic activity of Mrx2 wild type and mutants against compound nNF-C2; ( F ) ribbon representation of the crystal structure of Mrx2 (PDB 4ZIL), showing the localization of the Mrx2 mutations.

Article Snippet: Mycobacteria strains used in this study included: M. tuberculosis H37Rv (ATCC 27294 ) , M. tuberculosis H37Rv sigH null mutant (ST49; ( )), M. tuberculosis H37Rv mrx2 null mutant (B461c; ( )) a clinical Beijing M. tuberculosis strain (GC1237) which was isolated from a patient who was infected in the reported TB outbreak in Gran Canaria Islands (Spain) , two different multidrug resistant M. tuberculosis clinical strains isolated from TB patients at the Padova University Hospital (Padova, Italy), M ycobacterium abscessus ATCC19977, M ycobacterium aurum ATCC23366, Mycobacterium avium subsp. hominissuis ATCC 25291, Mycobacterium bovis BCG 1173 P2, M ycobacterium marinum ATCC BAA-535 and M ycobacterium smegmatis mc 2 155 .

Techniques: Activity Assay, In Vitro, Clone Assay, Expressing

Journal: Frontiers in Microbiology

Article Title: Genomic analysis of Mycobacterium brumae sustains its nonpathogenic and immunogenic phenotype

doi: 10.3389/fmicb.2022.982679

Figure Lengend Snippet: Safety profile of Mycobacterium brumae . Comparison of the pathogenicity and toxicity between M . brumae and M . bovis BCG Connaught. Summary of the results obtained in different in vitro studies infecting macrophages and bladder cancer cell lines , and in vivo studies in different animal models: intrahemacoelical infection of Galleria mellonella , intravesical instillations in orthotopic murine model of bladder cancer ( , , ), and intravenous infection in SCID mice . Blue rising arrow (high cell immune infiltration).

Article Snippet: The presence or absence of each gene was evaluated and compared in the M . bovis BCG Connaught and M . brumae ATCC 51384 T strains.

Techniques: In Vitro, In Vivo, Infection

Journal: Frontiers in Microbiology

Article Title: Genomic analysis of Mycobacterium brumae sustains its nonpathogenic and immunogenic phenotype

doi: 10.3389/fmicb.2022.982679

Figure Lengend Snippet: Genome features.

Article Snippet: The presence or absence of each gene was evaluated and compared in the M . bovis BCG Connaught and M . brumae ATCC 51384 T strains.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Genomic analysis of Mycobacterium brumae sustains its nonpathogenic and immunogenic phenotype

doi: 10.3389/fmicb.2022.982679

Figure Lengend Snippet: Genes presence or absence analysis. Venn diagram showing the comparison of the presence/absence and abundance of M. tuberculosis virulence-associated genes (A) , antigens recognized by T cells (B) , and antigens recognized by B cells (C) in the M. bovis BCG Connaught and M. brumae genomes.

Article Snippet: The presence or absence of each gene was evaluated and compared in the M . bovis BCG Connaught and M . brumae ATCC 51384 T strains.

Techniques:

Journal: Frontiers in Microbiology

Article Title: Genomic analysis of Mycobacterium brumae sustains its nonpathogenic and immunogenic phenotype

doi: 10.3389/fmicb.2022.982679

Figure Lengend Snippet: Immunogenic capacity of Mycobacterium brumae compared to Mycobacterium bovis BCG Connaught. Summary of the results obtained in studies treating with both mycobacteria J774 macrophages cell line , bladder cancer cell lines , human peripheral mononuclear cells , and murine bone-marrow macrophages (unpublished); and after treating intravesically with each mycobacteria bladder cancer tumor-bearing mice ( , , ). Blue rising arrow indicates higher than M . bovis BCG Connaught, red falling arrow indicates lower than M . bovis BCG Connaught and the equals symbol indicates similar to M . bovis BCG Connaught.

Article Snippet: The presence or absence of each gene was evaluated and compared in the M . bovis BCG Connaught and M . brumae ATCC 51384 T strains.

Techniques:

Journal: PLoS ONE

Article Title: The Gut as Reservoir of Antibiotic Resistance: Microbial Diversity of Tetracycline Resistance in Mother and Infant

doi: 10.1371/journal.pone.0021644

Figure Lengend Snippet: Part of the sequenced fosmid-insert in the infant Tc r metagenome (GU951538; 1–3095) is compared with the most similar homologous sequences in current databases, from S. gallolyticus subsp. gallolyticus strain UCN34 (FN597254; minus strand 564709–589025 and 52545–55152), S. gallolyticus subsp. pasteurianus NTUH7421 (AY894138; minus strand 1–4107) and S. bovis ATCC700338 (AEEL01000024; 112037–123526, AEEL01000025; 1–6541, AEEL01000026; 1–1578, AEEL01000027; 1–3192, AEEL01000028; 37166–39101). The relationships between sequences are shown as percentage identity at nucleotide level, calculated with the EMBOSS program Needle ( http://www.ebi.ac.uk/Tools/emboss/align/index.html ). Light gray arrows represent ORFs with homology to ORFs from Tn 916 /Tn 1545 -like conjugative transposons: orf 5–10, orf 12–24, tet (M), an excisionase ( xis ) and an integrase ( int ) of Tn 6079 . Dark gray arrows illustrate ORFs thay may be of plasmid origin: tet (L), pre/mob and rep . Black arrows illustrate ORFs that appear to be inserted by the two identical IS 1216 -like elements: first IS 1216 , erm (T), erm (T) leader sequence and second IS 1216 . rmp G and rmp F were predicted to encode L33 and L32 of the 50S ribosomal subunit. The functions of the predicted ORFs, mid12_1 , mid12_5 and mid12_7 are unknown. his S: histidyl-tRNA synthetase. IS Sdy1 : putative transposase.

Article Snippet: The reverse primer used in this PCR was specifically designed to target a sequence just downstream of the rpmF ORF that was present in the sequenced fosmid and in S. gallolyticus subsp. pasteurianus strain NTUH 7421 and S. bovis ATCC 7000338 (AEEL01000028) but not in S. gallolyticus subsp. gallolyticus strain UCN34.

Techniques: Plasmid Preparation, Sequencing