Journal: Cell

Article Title: Discovery of natural-product-derived sequanamycins as potent oral anti-tuberculosis agents

doi: 10.1016/j.cell.2023.01.043

Figure Lengend Snippet: In vivo activity of SEQ-9 in the lungs of Mtb-infected BALB/c mice after treatment with SEQ-9 Histograms represent the mean lung log 10 CFU counts (5–10 mice per group). Error bars in all panels represent SEM. The number following the compound abbreviation indicates the dose in mg/kg/day (unless otherwise indicated, ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001, compared with no treatment (one-way ANOVA). (A) SEQ-9 dose-ranging in an acute TB mouse model. CFU counts after 4 weeks of treatment. Day 3 indicates lung CFU counts at treatment initiation (3 days after infection). (B) SEQ-9 dose-ranging in the mouse model of chronic TB: CFU counts after 4 weeks of treatment. Day 3 indicates lung CFU 3 days after infection. Day 31 indicates lung CFU counts at treatment initiation (28 days after infection). A PEG200/Tween80/citrate buffer (PEG-Tw) 100 mM (70/5/27) pH 4 formulation was used for SEQ-9, whereas MCT was used for rifampicin (RIF) and linezolid (LIN) ( ∗ p < 0.05 and ∗∗ p < 0.01, compared with no treatment. Kruskal-Walllis test). (C) SEQ-9 dose-ranging in short (highly) acute mouse model of TB. Day 1 indicates lung CFU counts at treatment initiation (1 day after infection). SEQ-9 (Lip/2) and control (day 11 Lip/2) groups are from an independent study using a lipidic formulation described in Figure S8 and Table S7 and were compared by Student’s t test: ###p < 0.001.

Article Snippet: A mid log phase Mtb H37Rv culture was incubated overnight in Middlebrook 7H9 broth (supplemented with + 10 % OADC + 0.05% Tween80 + 0.4% glycérol) with subinhibitory concentrations of clarithromycin or SEQ-9 (0.125μg/ml and 0.25μg/ml).

Techniques: In Vivo, Activity Assay, Infection

Journal: Cell

Article Title: Discovery of natural-product-derived sequanamycins as potent oral anti-tuberculosis agents

doi: 10.1016/j.cell.2023.01.043

Figure Lengend Snippet: SEQ-9 kill kinetics of Mtb H37Rv in 7H9 broth culture, related to Figure 5 Bacterial cultures with various SEQ-9 concentrations or 1% DMSO were followed during 11 days. For each time, CFU count was assessed by plating on 7H11 agar plates. The number following the compound abbreviation indicates the concentration in μM.

Article Snippet: A mid log phase Mtb H37Rv culture was incubated overnight in Middlebrook 7H9 broth (supplemented with + 10 % OADC + 0.05% Tween80 + 0.4% glycérol) with subinhibitory concentrations of clarithromycin or SEQ-9 (0.125μg/ml and 0.25μg/ml).

Techniques: Concentration Assay

Journal: Cell

Article Title: Discovery of natural-product-derived sequanamycins as potent oral anti-tuberculosis agents

doi: 10.1016/j.cell.2023.01.043

Figure Lengend Snippet:

Article Snippet: A mid log phase Mtb H37Rv culture was incubated overnight in Middlebrook 7H9 broth (supplemented with + 10 % OADC + 0.05% Tween80 + 0.4% glycérol) with subinhibitory concentrations of clarithromycin or SEQ-9 (0.125μg/ml and 0.25μg/ml).

Techniques: Mutagenesis, Recombinant, Cell Culture, Methylation, Amplification, Software, Microscopy, RNA Extraction, Real-time Polymerase Chain Reaction

Journal: Journal of Applied Microbiology

Article Title: Comparative survival of environmental and clinical Mycobacterium abscessus isolates in a variety of diverse host cells

doi: 10.1111/jam.15416

Figure Lengend Snippet: Isolate panel characteristics. (a) Phylogenetic analyses of isolates used in this study. A phylogeny based on the multiple sequence alignment of partial rpoB sequences inferred between isolate relationships was obtained using the neighbor‐joining method and observed SNPs. The scale bar represents a 4 SNP difference in nucleotide sequences. (b) Colony morphology of the isolate panel after streaking for isolation on Middlebrook 7H10 agar plates and viewed under a dissecting scope. Total magnification = 40X. (c) Pellicle formation was assessed after incubation for 23 days in Middlebrook 7H9 broth culture. Isolates that produced observable bacterial aggregation at the air‐liquid interface of the culture were determined to have formed a pellicle. (d) TLC of total lipid extracts from the isolate panel. 25 μl of each sample were spotted onto 13 × 10 cm analytical TLC plates and eluted in 65:25:4 HCCl 3 :MeOH:H 2 O. Total lipids were visualized with CuSO4. Isolate 4 = Arthrobacter sp

Article Snippet: Amoebae in the 1 h wells were lysed using 0.025% SDS and 7H9 plating solution (2.35 gl −1 Middlebrook 7H9 base, 0.1% Tween 80, 10% OADC supplement).

Techniques: Sequencing, Isolation, Incubation, Produced

Journal: Journal of Applied Microbiology

Article Title: Comparative survival of environmental and clinical Mycobacterium abscessus isolates in a variety of diverse host cells

doi: 10.1111/jam.15416

Figure Lengend Snippet: Growth of the isolate panel in culture media and a variety of host cells. (a) Growth curve of the isolate panel in Middlebrook 7H9 broth incubated at 37°C. CFU were quantified at 1, 24, 48 and 96 h post inoculation. n = 3 independent experiments. * p < 0.05; ** p < 0.005; **** p < 1 × 10 −16 . (b) Co‐culture of the isolate panel with Acanthamoeba lenticulata . 1 × 10 5 A . lenticulata and a MOI 1:1 were used. CFU were quantified at 1, 24, 48 and 96 h post infection. Amoebae co‐culture was performed at 22°C. At least 3 independent experiments were performed for all isolates. * p < 0.05 (comparing Isolate 1 vs 2); **** p < 1 × 10 −16 . (c) The isolate panel was co‐cultured with 2 × 10 5 BEAS‐2B airway epithelial cells using a MOI 1:1. CFU were quantified at 1, 24, 48 and 96 h post inoculation. ** p < 0.005, **** p < 1 × 10 −16 . (d) 1 × 10 6 primary human alveolar macrophages were co‐cultured ex vivo with the isolate panel using a MOI 1:1. CFU were quantified at 1, 24, 48 and 96 h post inoculation. * p < 0.05; ** p < 0.005; **** p < 1 × 10 −16 . Isolate 4 = Arthrobacter sp .

Article Snippet: Amoebae in the 1 h wells were lysed using 0.025% SDS and 7H9 plating solution (2.35 gl −1 Middlebrook 7H9 base, 0.1% Tween 80, 10% OADC supplement).

Techniques: Incubation, Co-Culture Assay, Infection, Cell Culture, Ex Vivo