Journal: Science Advances

Article Title: A streamlined CRISPR-based test for tuberculosis detection directly from sputum

doi: 10.1126/sciadv.adx2067

Figure Lengend Snippet: ( A ) Cas13a detecting BCG genomic DNA spiked into sputum:TE matrix and Cas12a detecting internal control. Sp is sputum, and gBCG is genomic BCG. Units are genomes per microliter. Cas13a reporter is FAM, and Cas12a reporter is HEX. ( B ) Comparison of fresh Cas13a (left) or lyophilized Cas13a (right) assays detecting synthetic internal control (IC) and BCG genomic DNA. Synthetic IC and genomic BCG were mixed at a concentration of 10 4 and 10 3 copies per microliter, respectively. ( C ) Lateral flow readout of Cas13a detecting dual targets IS6110 + IS1081 (top) and Cas12a detecting internal control (bottom) in BCG spiked into sputum:TE matrix. Appearance of the top line is a positive readout. Faint lines are interpreted as negative. Disappearance of the bottom line is not necessary. ( D ) Range finding experiment demonstrating the dynamic range of fresh Cas13a dual-detection assay. Cas13a detecting BCG and Cas12a detecting internal control. BCG culture [left; P adj (left to right) is <0.001, <0.001, <0.001, <0.01, <0.05, n.s., n.s., and n.s.] and BCG spiked into sputum:TE [right; P adj (left to right) is <0.0001, <0.0001, <0.0001, <0.05, n.s., n.s., and n.s.]. gBCG is genomic BCG and units are genomes per microliter. All other units are colony-forming units (CFU) per milliliter. ( E ) Testing fresh Cas13a and Cas12a coupled assay against TB-negative clinical sputum samples. Numbers indicate patient number. All samples are n.s. except Sp-17, where P = 0.015. ( F ) LoD for H37Rv. LoD = 69.0 (51.0 to 86.9). ( G ) LoD for BCG. LoD = 80.5 (59.4 to 101.6). In (A), (B), (D), and (E), error bars: SD based on n = 3. (C) A single representative sample. (F) n = 20 and (G) n = 21. In (D) and (E), one-way ANOVA with Dunnett’s test was performed, and P adj values were calculated compared to NTC that contained water. a.u., arbitrary units.

Article Snippet: BCG genomic DNA was ordered from ATCC (catalog no. 35734D-2) and quantified with dPCR.

Techniques: Control, Comparison, Concentration Assay, Detection Assay

Journal: BMC Microbiology

Article Title: One-pot, uracil-DNA-glycosylase-aided, and multi-indicator nanobiosensor detection platform for identifying Brucella melitensis from the Brucella spp

doi: 10.1186/s12866-025-04189-9

Figure Lengend Snippet: Analytical specificity of mMCUDA assay using an AuNPs-LFA biosensor. The mMCUDA assay was implemented using nucleic acids extracted from various pathogens as templates. Strips 1–5 correspond to plasmids, B. melitensis (M5), B. melitensis (16 M, NCTC 10094), B. melitensis (Ether, NCTC 10509), and B. melitensis (standard material). Strips 6–11 correspond to B. melitensis (isolates). Strip 12–15 correspond to B. abortus (544, NCTC 10093), B. abortus (A19), B. abortus (isolate), and B. abortus (standard material). Strips 16–18 correspond to B. suis (1330, NCTC 10316), B. suis (S2), and B. suis (standard material). Strips 19–38 correspond to M. tuberculosis H37Rv (ATCC 27294), M. tuberculosis H37Ra (ATCC 25177), M. tuberculosis isolate (GZCDC), M. malmoense (ATCC 29571), M. bovis (ATCC19210), Listeria monocytogenes (GZCDC), Staphylococcus aureus (GZCDC), Streptococcus pneumoniae (GZCDC), Escherichia coli (GZCDC), Klebsiella pneumoniae (GZCDC), Pseudomonas aeruginosa (GZCDC), Shigella spp. (GZCDC), Streptococcus suis (GZCDC), Orientia tsutsugamushi (GZCDC), M. leprae (GZCDC), Salmonella spp. (GZCDC), Bordetella pertussis (GZCDC), Neisseria meningitidis (GZCDC), Human cytomegalovirus (BNCC), and Influenza virus (GZCDC). Strips 39-40 correspond to negative control (environmental samples in the experiment) and blank control (DNase/RNase-free water). NCTC, National Collection of Type Cultures; BNCC, BeNa Culture Collection; GZCDC, Guizhou Provincial Center for Disease Control and Prevention; ATCC, American Type Culture Collection; mMCUDA, m ultiplex M CDA C ombined with dUTP-aided U DGase and D etector-based A uNPs-LFA biosensor; MCDA, multiple cross-displacement amplification; AuNPs-LFA, gold nanoparticles-based lateral flow assay; TL1, test line 1; TL2, test line 2; CL, control line

Article Snippet: Strips 19–38 correspond to M. tuberculosis H37Rv (ATCC 27294), M. tuberculosis H37Ra (ATCC 25177), M. tuberculosis isolate (GZCDC), M. malmoense (ATCC 29571), M. bovis (ATCC19210), Listeria monocytogenes (GZCDC), Staphylococcus aureus (GZCDC), Streptococcus pneumoniae (GZCDC), Escherichia coli (GZCDC), Klebsiella pneumoniae (GZCDC), Pseudomonas aeruginosa (GZCDC), Shigella spp. (GZCDC), Streptococcus suis (GZCDC), Orientia tsutsugamushi (GZCDC), M. leprae (GZCDC), Salmonella spp. (GZCDC), Bordetella pertussis (GZCDC), Neisseria meningitidis (GZCDC), Human cytomegalovirus (BNCC), and Influenza virus (GZCDC).

Techniques: Stripping Membranes, Virus, Negative Control, Environmental Sampling, Control, Amplification, Lateral Flow Assay

Journal: The Journal of Biological Chemistry

Article Title: Mycobacterium tuberculosis Directs T Helper 2 Cell Differentiation by Inducing Interleukin-1β Production in Dendritic Cells

doi: 10.1074/jbc.M112.375154

Figure Lengend Snippet: ESAT-6 induces Th2 cell differentiation via IL-1β production. A , IL-4 production in DCs co-cultured with naïve OT-II CD4 + T cells after infection with H37Rv or BCG in the presence or absence of anti-IL-1β. B , IL-4 production in DCs co-cultured with naïve OT-II CD4 + T cells after infection with H37Rv or mutant strains (H37RvΔRD1, H37RvΔESAT-6, and Comp ESAT-6) in the presence or absence of anti-IL-1β. IL-4 levels were higher in cultures with H37Rv or Comp ESAT-6 compared with cultures with BCG or other mutant strains ( p < 0.003). C , Western blot analysis of STAT6, pSTAT6, STAT4, and pSTAT4 proteins from DCs infected with H37Rv or H37RvΔESAT-6 and co-cultured with OT-II CD4 + T cells, to show modulated Th1 and Th2 responses in the presence or absence of anti-IL-1β. The results shown are representative of at least four independent experiments. Error bars , ±S.D.

Article Snippet: BCG, H37RvΔRD-1, H37RvΔESAT-6, and ESAT-6-complemented H37RvΔESAT-6 were a kind gift from Prof. David Sherman (SBRI, Seattle, WA).

Techniques: Cell Differentiation, Cell Culture, Infection, Mutagenesis, Western Blot

Journal: The Journal of Biological Chemistry

Article Title: Mycobacterium tuberculosis Directs T Helper 2 Cell Differentiation by Inducing Interleukin-1β Production in Dendritic Cells

doi: 10.1074/jbc.M112.375154

Figure Lengend Snippet: H37Rv induces significantly higher Th2 responses and the proinflammatory cytokine IL-1β compared with BCG or H37RvΔESAT-6. A , cytokine levels in lung homogenates of mice at different time points after infection with H37Rv or BCG. B , cytokine levels in lung homogenates of mice at different time points after infection with H37Rv or mutant strain H37RvΔESAT-6. Statistical significance between different strains was determined by one-way ANOVA: *, p < 0.01; **, p < 0.017; ***, p < 0.001. The results shown are representative of at least three independent experiments with six mice within each group. Error bars , ±S.D.

Article Snippet: BCG, H37RvΔRD-1, H37RvΔESAT-6, and ESAT-6-complemented H37RvΔESAT-6 were a kind gift from Prof. David Sherman (SBRI, Seattle, WA).

Techniques: Infection, Mutagenesis

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The liposome of trehalose dimycolate extracted from M. bovis BCG induces antitumor immunity via the activation of dendritic cells and CD8 + T cells

doi: 10.1007/s00262-021-02870-2

Figure Lengend Snippet: Isolation of TDM from M. bovis BCG Connaught and the preparation of liposomes. a Chemical structure of trehalose dimycolate (TDM). TDM from Mycobacterium. bovis BCG Connaught has α- and keto-mycolic acid, but no methoxy mycolic acid. b Thin-layer chromatography of TDM from M. bovis BCG Connaught. Left lane: chloroform/methanol (C/M) extraction. Right lane: Purified TDM. c The dia., polydispersity index (PdI), and zeta potential of the cationic liposome TDM (Lip-TDM) and the empty liposome (Lip-Con) measured by a ZETASIZERNano. Each value is the average of three measurements

Article Snippet: Isolation and purification of TDM from heat-killed BCG cells by solvent fractionation M. bovis BCG Connaught was kindly provided by Japan BCG Laboratory (Kiyose, Japan).

Techniques: Isolation, Liposomes, Thin Layer Chromatography, Extraction, Purification, Zeta Potential Analyzer

Journal: Cancer Immunology, Immunotherapy : CII

Article Title: The liposome of trehalose dimycolate extracted from M. bovis BCG induces antitumor immunity via the activation of dendritic cells and CD8 + T cells

doi: 10.1007/s00262-021-02870-2

Figure Lengend Snippet: Antitumor effect of Lip-TDM by subcutaneous administration. a Tumor-bearing mice were treated subcutaneously (s.c.) with Lip-Con, Lip-TDM, or BCG 3 h later (day 0) and on days 3, 5, and 7. b–d Wild type (WT) BL6 mice were inoculated with MB49 cells, MC38 cells, or B16F10 cells. e, f Lip-TDM was injected into the tumor site in the mice bearing MB49 cells on day 0, 1, 3, or 7, followed by an injection every 2 days. Tumor volumes were monitored 2 × /week and are presented as the mean ± SEM (n = 5–7 per group). *p < 0.05, **p < 0.01 by Mann–Whitney U test. In vivo experiments were performed at least twice

Article Snippet: Isolation and purification of TDM from heat-killed BCG cells by solvent fractionation M. bovis BCG Connaught was kindly provided by Japan BCG Laboratory (Kiyose, Japan).

Techniques: Injection, MANN-WHITNEY, In Vivo

Journal: Diseases

Article Title: Clinical Impact of the Increase in Immunosuppressive Cell-Related Gene Expression in Urine Sediment during Intravesical Bacillus Calmette-Guérin

doi: 10.3390/diseases7020044

Figure Lengend Snippet: Time-course changes in blood and urine clinical parameters before, during, and after intravesical BCG.

Article Snippet: All patients received an induction course of intravesical BCG (BCG Tokyo 172 strain; Japan BCG Laboratory Tokyo, Japan) once weekly for 8 weeks.

Techniques:

Journal: Diseases

Article Title: Clinical Impact of the Increase in Immunosuppressive Cell-Related Gene Expression in Urine Sediment during Intravesical Bacillus Calmette-Guérin

doi: 10.3390/diseases7020044

Figure Lengend Snippet: Time-course changes in mRNA expression of immune cell markers in urine sediments before, during, and after intravesical BCG.

Article Snippet: All patients received an induction course of intravesical BCG (BCG Tokyo 172 strain; Japan BCG Laboratory Tokyo, Japan) once weekly for 8 weeks.

Techniques: Expressing, RNA Expression, TaqMan Assay

Journal: Diseases

Article Title: Clinical Impact of the Increase in Immunosuppressive Cell-Related Gene Expression in Urine Sediment during Intravesical Bacillus Calmette-Guérin

doi: 10.3390/diseases7020044

Figure Lengend Snippet: The prognostic factors for recurrence in 24 patients treated with intravesical BCG.

Article Snippet: All patients received an induction course of intravesical BCG (BCG Tokyo 172 strain; Japan BCG Laboratory Tokyo, Japan) once weekly for 8 weeks.

Techniques: Expressing

Journal: Diseases

Article Title: Clinical Impact of the Increase in Immunosuppressive Cell-Related Gene Expression in Urine Sediment during Intravesical Bacillus Calmette-Guérin

doi: 10.3390/diseases7020044

Figure Lengend Snippet: Intravesical recurrence-free survival curves. ( A ) Stratification according to the number of upregulated genes in urine sediment after intravesical BCG. The log-rank test was used for the comparison between 0–1 (n = 14) and 2–4 (n = 10). ( B ) Stratification according to the CUETO recurrence score. The log-rank test was used for the comparison between 0–6 (n = 9) and 7–16 (n = 15). HR, hazard ratio; CI, confidence interval.

Article Snippet: All patients received an induction course of intravesical BCG (BCG Tokyo 172 strain; Japan BCG Laboratory Tokyo, Japan) once weekly for 8 weeks.

Techniques: Comparison