streptococcus pneumoniae  (ATCC)


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    Streptococcus pneumoniae CIP 104225
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    Structured Review

    ATCC streptococcus pneumoniae
    Effects of CD137 signaling modulation on survival rates of septic mice induced by G + or G − bacteria. (A) CD137 +/+ mice were injected with 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either G + bacteria (7.5 × 10 8 S. aureus , 1 × 10 4 S. <t>pneumoniae</t> , or 8 × 10 7 E. faecalis organisms) (left graphs) or G − bacteria (1 × 10 7 E. coli , 6 × 10 8 P. aeruginosa , or 2 × 10 6 S . Typhimurium organisms) (right graphs). (B) CD137 −/− mice were administered 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either 3.5 × 10 8 S. aureus or 2 × 10 7 E. coli organisms. Mouse survival was determined for 7 days. Each group contained 10 to 15 mice, and results of 2 or 3 different experiments were pooled. #, P

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    Images

    1) Product Images from "CD137 Expressed on Neutrophils Plays Dual Roles in Antibacterial Responses against Gram-Positive and Gram-Negative Bacterial Infections"

    Article Title: CD137 Expressed on Neutrophils Plays Dual Roles in Antibacterial Responses against Gram-Positive and Gram-Negative Bacterial Infections

    Journal: Infection and Immunity

    doi: 10.1128/IAI.00115-13

    Effects of CD137 signaling modulation on survival rates of septic mice induced by G + or G − bacteria. (A) CD137 +/+ mice were injected with 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either G + bacteria (7.5 × 10 8 S. aureus , 1 × 10 4 S. pneumoniae , or 8 × 10 7 E. faecalis organisms) (left graphs) or G − bacteria (1 × 10 7 E. coli , 6 × 10 8 P. aeruginosa , or 2 × 10 6 S . Typhimurium organisms) (right graphs). (B) CD137 −/− mice were administered 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either 3.5 × 10 8 S. aureus or 2 × 10 7 E. coli organisms. Mouse survival was determined for 7 days. Each group contained 10 to 15 mice, and results of 2 or 3 different experiments were pooled. #, P
    Figure Legend Snippet: Effects of CD137 signaling modulation on survival rates of septic mice induced by G + or G − bacteria. (A) CD137 +/+ mice were injected with 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either G + bacteria (7.5 × 10 8 S. aureus , 1 × 10 4 S. pneumoniae , or 8 × 10 7 E. faecalis organisms) (left graphs) or G − bacteria (1 × 10 7 E. coli , 6 × 10 8 P. aeruginosa , or 2 × 10 6 S . Typhimurium organisms) (right graphs). (B) CD137 −/− mice were administered 200 μg of 3E1, TKS-1, or rat IgG. After 24 h, they were inoculated i.p. with either 3.5 × 10 8 S. aureus or 2 × 10 7 E. coli organisms. Mouse survival was determined for 7 days. Each group contained 10 to 15 mice, and results of 2 or 3 different experiments were pooled. #, P

    Techniques Used: Mouse Assay, Injection

    CD137-deficient mice are resistant to infection with G − bacteria but susceptible to infection with G + bacteria. BALB/c CD137 −/− and CD137 +/+ littermates were inoculated i.p. with either G + bacteria (5 × 10 8 S. aureus , 5 × 10 3 S. pneumoniae , or 8 × 10 7 E. faecalis organisms) (A) or G − bacteria (1 × 10 7 E. coli , 6 × 10 8 P. aeruginosa , or 2 × 10 6 S . Typhimurium organisms) (B), and mouse survival was monitored for 7 days. Each group contained 10 to 15 mice, and the results of 2 or 3 different experiments were pooled. *, P
    Figure Legend Snippet: CD137-deficient mice are resistant to infection with G − bacteria but susceptible to infection with G + bacteria. BALB/c CD137 −/− and CD137 +/+ littermates were inoculated i.p. with either G + bacteria (5 × 10 8 S. aureus , 5 × 10 3 S. pneumoniae , or 8 × 10 7 E. faecalis organisms) (A) or G − bacteria (1 × 10 7 E. coli , 6 × 10 8 P. aeruginosa , or 2 × 10 6 S . Typhimurium organisms) (B), and mouse survival was monitored for 7 days. Each group contained 10 to 15 mice, and the results of 2 or 3 different experiments were pooled. *, P

    Techniques Used: Mouse Assay, Infection

    2) Product Images from "Zincophorin – biosynthesis in Streptomyces griseus and antibiotic properties"

    Article Title: Zincophorin – biosynthesis in Streptomyces griseus and antibiotic properties

    Journal: GMS Infectious Diseases

    doi: 10.3205/id000026

    Dose dependence of the effect of zincophorin on growth and biofilm formation of S. pneumoniae DSM20566. Mean values of the inhibitory effect in % with standard deviation of at least 3 tests each with 2 parallels are shown.
    Figure Legend Snippet: Dose dependence of the effect of zincophorin on growth and biofilm formation of S. pneumoniae DSM20566. Mean values of the inhibitory effect in % with standard deviation of at least 3 tests each with 2 parallels are shown.

    Techniques Used: Standard Deviation

    Antibacterial activity of zincophorin, daptomycin, and imipenem. Mean MIC and MBIC values in μM with standard deviation determined for three reference strains and four clinical isolates of S. pneumoniae in microtiter broth dilution and biofilm assay, respectively, are shown.
    Figure Legend Snippet: Antibacterial activity of zincophorin, daptomycin, and imipenem. Mean MIC and MBIC values in μM with standard deviation determined for three reference strains and four clinical isolates of S. pneumoniae in microtiter broth dilution and biofilm assay, respectively, are shown.

    Techniques Used: Activity Assay, Standard Deviation, Biofilm Production Assay

    3) Product Images from "Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection"

    Article Title: Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1303090

    Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p
    Figure Legend Snippet: Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p

    Techniques Used: Inhibition, Infection, Cell Culture, Isolation, Expressing, Western Blot, Mouse Assay, Immunoprecipitation, Activation Assay, Enzyme-linked Immunosorbent Assay, T-Test

    Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p
    Figure Legend Snippet: Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p

    Techniques Used: Inhibition, Infection, Mouse Assay, Injection

    Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p
    Figure Legend Snippet: Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p
    Figure Legend Snippet: STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Real-time Polymerase Chain Reaction

    Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p
    Figure Legend Snippet: Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Enzyme-linked Immunosorbent Assay

    Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p
    Figure Legend Snippet: Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p

    Techniques Used: Infection, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p
    Figure Legend Snippet: Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    4) Product Images from "Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection"

    Article Title: Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1303090

    Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p
    Figure Legend Snippet: Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p

    Techniques Used: Inhibition, Infection, Cell Culture, Isolation, Expressing, Western Blot, Mouse Assay, Immunoprecipitation, Activation Assay, Enzyme-linked Immunosorbent Assay, T-Test

    Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p
    Figure Legend Snippet: Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p

    Techniques Used: Inhibition, Infection, Mouse Assay, Injection

    Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p
    Figure Legend Snippet: Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p
    Figure Legend Snippet: STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Real-time Polymerase Chain Reaction

    Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p
    Figure Legend Snippet: Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Enzyme-linked Immunosorbent Assay

    Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p
    Figure Legend Snippet: Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p

    Techniques Used: Infection, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p
    Figure Legend Snippet: Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    5) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    6) Product Images from "Aminomethyl Spectinomycins as Novel Therapeutics for Drug Resistant Respiratory Tract and Sexually Transmitted Bacterial Infections"

    Article Title: Aminomethyl Spectinomycins as Novel Therapeutics for Drug Resistant Respiratory Tract and Sexually Transmitted Bacterial Infections

    Journal: Science translational medicine

    doi: 10.1126/scitranslmed.3010572

    Compound 1 modeled into the bacterial ribosome of S. pneumoniae, which shows the aryl side chain positioned in a side pocket adjacent to RpsE loop. ( A ) Structural variances between the E. coli and S. pneumoniae are highlighted in red within the loop of
    Figure Legend Snippet: Compound 1 modeled into the bacterial ribosome of S. pneumoniae, which shows the aryl side chain positioned in a side pocket adjacent to RpsE loop. ( A ) Structural variances between the E. coli and S. pneumoniae are highlighted in red within the loop of

    Techniques Used:

    7) Product Images from "Large scale multiplex PCR improves pathogen detection by DNA microarrays"

    Article Title: Large scale multiplex PCR improves pathogen detection by DNA microarrays

    Journal: BMC Microbiology

    doi: 10.1186/1471-2180-9-1

    Specific detection of microbial DNA by LSplex amplification . Hybridization profiles generated by analysis of LSplex amplified products shown as columns ( S. aureus, E. coli, S. pneumonia, E. faecalis, P. mirabilis, S. epidermidis, K. pneumoniae, C. albicans and P. aeruginosa ). Each row represents an individual capture probe of the microarray, grouped by species or genus specific regions (see Additional file 2 ) as indicated in the left column. The boxes represent the positive hybridization signal of bacterial DNA (in colour) or absence of hybridisation (in white) with individual capture probes.
    Figure Legend Snippet: Specific detection of microbial DNA by LSplex amplification . Hybridization profiles generated by analysis of LSplex amplified products shown as columns ( S. aureus, E. coli, S. pneumonia, E. faecalis, P. mirabilis, S. epidermidis, K. pneumoniae, C. albicans and P. aeruginosa ). Each row represents an individual capture probe of the microarray, grouped by species or genus specific regions (see Additional file 2 ) as indicated in the left column. The boxes represent the positive hybridization signal of bacterial DNA (in colour) or absence of hybridisation (in white) with individual capture probes.

    Techniques Used: Amplification, Hybridization, Generated, Microarray

    8) Product Images from "Conserved Streptococcus pneumoniae Spirosomes Suggest a Single Type of Transformation Pilus in Competence"

    Article Title: Conserved Streptococcus pneumoniae Spirosomes Suggest a Single Type of Transformation Pilus in Competence

    Journal: PLoS Pathogens

    doi: 10.1371/journal.ppat.1004835

    Pneumococcal transformation pilus controversy: Long, T4P-like transformation pilus versus short, ‘plaited’ polymers. (A) Long, T4P-like transformation pilus protruding on the surface of competence-induced S . pneumoniae strain R1501 [ 10 ]. (B) High-resolution structural model of a T2SS-peudopilus in side (left) and top (right) views [ 15 ], visualized in PyMOL (Schrödinger). (C) Scaled electron density reprojections of the T2SS-pseudopilus model (left), compared to class averages of the long T4P-like transformation pilus reported in [ 10 ] (center) and the short, ‘plaited’ filaments reported in [ 8 ] (right). Scale bars 5 nm. (D) Coexistence of the T4P-like pilus (black arrowhead) and ‘plaited’ filaments (white arrowhead) in competence-induced S . pneumoniae culture. (E) Immunogold labeling of major pilin ComGC in a strain carrying an additional FLAG-tagged ectopic copy of the comGC gene. A T4P-like pilus and a ‘plaited’ filament are indicated by black and white arrowheads, respectively. (F) and (G) Unabolished release of ‘plaited’ filaments in non-competent, pilus-deficient ΔcomGB [ 17 ] and ΔcomGA S . pneumoniae , respectively.
    Figure Legend Snippet: Pneumococcal transformation pilus controversy: Long, T4P-like transformation pilus versus short, ‘plaited’ polymers. (A) Long, T4P-like transformation pilus protruding on the surface of competence-induced S . pneumoniae strain R1501 [ 10 ]. (B) High-resolution structural model of a T2SS-peudopilus in side (left) and top (right) views [ 15 ], visualized in PyMOL (Schrödinger). (C) Scaled electron density reprojections of the T2SS-pseudopilus model (left), compared to class averages of the long T4P-like transformation pilus reported in [ 10 ] (center) and the short, ‘plaited’ filaments reported in [ 8 ] (right). Scale bars 5 nm. (D) Coexistence of the T4P-like pilus (black arrowhead) and ‘plaited’ filaments (white arrowhead) in competence-induced S . pneumoniae culture. (E) Immunogold labeling of major pilin ComGC in a strain carrying an additional FLAG-tagged ectopic copy of the comGC gene. A T4P-like pilus and a ‘plaited’ filament are indicated by black and white arrowheads, respectively. (F) and (G) Unabolished release of ‘plaited’ filaments in non-competent, pilus-deficient ΔcomGB [ 17 ] and ΔcomGA S . pneumoniae , respectively.

    Techniques Used: Transformation Assay, Labeling

    Identification of the ‘plaited’ filaments as AdhE spirosomes. (A) Negatively stained purified ‘plaited’ filaments. (B) Left, coomassie stained SDS-PAGE on the purified filament fraction. Center, proteomic analysis of the predominant protein band; Posterior Error Probability (PEP) value for AdhE is 0. Right, Western blot validation using an anti-AdhE antibody (Agrisera). (C) Heterologous expression in E . coli and purification of AdhE S.pneumoniae . Left, coomassie stained SDS-PAGE of the affinity purified protein ( S2 Table ). Right, negatively stained electron microscopy of the purified protein with black arrowheads indicating spontaneously formed spirosomes. (D) Immunopurification of S . pneumoniae spirosomes from a strain carrying an additional competence-inducible FLAG-tagged ectopic copy of the adhE gene (SO007): left, coomassie-stained SDS-PAGE of the eluted fraction in CSP-induced and-uninduced cells (CSP, competence stimulating peptide); right, negative-stain EM on the eluted fraction (E) Top, AdhE domain organization; bottom, high-resolution structural model of a G . thermoglucosidasius spirosome [ 19 ], visualized in PyMOL (Schrödinger). The color of the individual protomers alternate between grayscale and domain-coded color representation; successive AdhE dimers alternate between ribbon and surface representation (F) A representative class average of the ‘plaited’ filaments compared to electron density reprojections of the G . thermoglucosidasius spirosome model [ 19 ]. Scale bars 5nm.
    Figure Legend Snippet: Identification of the ‘plaited’ filaments as AdhE spirosomes. (A) Negatively stained purified ‘plaited’ filaments. (B) Left, coomassie stained SDS-PAGE on the purified filament fraction. Center, proteomic analysis of the predominant protein band; Posterior Error Probability (PEP) value for AdhE is 0. Right, Western blot validation using an anti-AdhE antibody (Agrisera). (C) Heterologous expression in E . coli and purification of AdhE S.pneumoniae . Left, coomassie stained SDS-PAGE of the affinity purified protein ( S2 Table ). Right, negatively stained electron microscopy of the purified protein with black arrowheads indicating spontaneously formed spirosomes. (D) Immunopurification of S . pneumoniae spirosomes from a strain carrying an additional competence-inducible FLAG-tagged ectopic copy of the adhE gene (SO007): left, coomassie-stained SDS-PAGE of the eluted fraction in CSP-induced and-uninduced cells (CSP, competence stimulating peptide); right, negative-stain EM on the eluted fraction (E) Top, AdhE domain organization; bottom, high-resolution structural model of a G . thermoglucosidasius spirosome [ 19 ], visualized in PyMOL (Schrödinger). The color of the individual protomers alternate between grayscale and domain-coded color representation; successive AdhE dimers alternate between ribbon and surface representation (F) A representative class average of the ‘plaited’ filaments compared to electron density reprojections of the G . thermoglucosidasius spirosome model [ 19 ]. Scale bars 5nm.

    Techniques Used: Staining, Purification, SDS Page, Western Blot, Expressing, Affinity Purification, Electron Microscopy, Immu-Puri

    9) Product Images from "An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac"

    Article Title: An easy, rapid, and sensitive method for detection of drug-resistant influenza virus by using a sialidase fluorescent imaging probe, BTP3-Neu5Ac

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0200761

    Detection of drug-resistant viruses in the BTP3-based filter method. (A) 100 μM BTP3-Neu5Ac supplemented with or without 10 μM NAIs was added directly onto the influenza virus-loaded filter membranes. After 15 min at 56°C, BTP3 was visualized with a handheld UV flashlight and NAI susceptibility was evaluated by comparing the fluorescent images for NAI (-) and NAI-supplemented samples. NAI susceptibility of seven influenza virus strains was assessed for each NAI. (B) Fluorescence patterns of agents that may cause false-positive results. MuV, S . pneumoniae , and HPIV-1 with known titres were loaded onto the filter membranes and BTP3-Neu5Ac reaction was carried out as described in (A). OV, PV, ZV, and LV represent oseltamivir, peramivir, zanamivir, and laninamivir, respectively.
    Figure Legend Snippet: Detection of drug-resistant viruses in the BTP3-based filter method. (A) 100 μM BTP3-Neu5Ac supplemented with or without 10 μM NAIs was added directly onto the influenza virus-loaded filter membranes. After 15 min at 56°C, BTP3 was visualized with a handheld UV flashlight and NAI susceptibility was evaluated by comparing the fluorescent images for NAI (-) and NAI-supplemented samples. NAI susceptibility of seven influenza virus strains was assessed for each NAI. (B) Fluorescence patterns of agents that may cause false-positive results. MuV, S . pneumoniae , and HPIV-1 with known titres were loaded onto the filter membranes and BTP3-Neu5Ac reaction was carried out as described in (A). OV, PV, ZV, and LV represent oseltamivir, peramivir, zanamivir, and laninamivir, respectively.

    Techniques Used: Fluorescence

    10) Product Images from "Interferon-\u03b3 Production by Neutrophils during Bacterial Pneumonia in Mice"

    Article Title: Interferon-\u03b3 Production by Neutrophils during Bacterial Pneumonia in Mice

    Journal: American Journal of Respiratory and Critical Care Medicine

    doi: 10.1164/rccm.201004-0592OC

    S. pneumoniae and S. aureus Induce Production of IFN-γ in Neutrophils, whereas E. coli , P. aeruginosa , LPS, and LTA Do Not
    Figure Legend Snippet: S. pneumoniae and S. aureus Induce Production of IFN-γ in Neutrophils, whereas E. coli , P. aeruginosa , LPS, and LTA Do Not

    Techniques Used:

    IFN-γ is required for host defense against Streptococcus pneumoniae . ( A ) Representative immunomicrographs of neutrophils and neutrophil extracellular traps (NETs) after immunofluorescence and 4′,6-diamidino-2-phenylindole staining to detect
    Figure Legend Snippet: IFN-γ is required for host defense against Streptococcus pneumoniae . ( A ) Representative immunomicrographs of neutrophils and neutrophil extracellular traps (NETs) after immunofluorescence and 4′,6-diamidino-2-phenylindole staining to detect

    Techniques Used: Immunofluorescence, Staining

    Streptococcus pneumoniae and Staphylococcus aureus induced production of IFN-γ in lung neutrophils by 24 hours, whereas Escherichia coli , Pseudomonas aeruginosa , LPS, and lipoteichoic acid (LTA) did not. ( A ) Representative flow cytometric analyses
    Figure Legend Snippet: Streptococcus pneumoniae and Staphylococcus aureus induced production of IFN-γ in lung neutrophils by 24 hours, whereas Escherichia coli , Pseudomonas aeruginosa , LPS, and lipoteichoic acid (LTA) did not. ( A ) Representative flow cytometric analyses

    Techniques Used: Flow Cytometry

    11) Product Images from "Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection"

    Article Title: Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1303090

    Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p
    Figure Legend Snippet: Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p

    Techniques Used: Inhibition, Infection, Cell Culture, Isolation, Expressing, Western Blot, Mouse Assay, Immunoprecipitation, Activation Assay, Enzyme-linked Immunosorbent Assay, T-Test

    Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p
    Figure Legend Snippet: Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p

    Techniques Used: Inhibition, Infection, Mouse Assay, Injection

    Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p
    Figure Legend Snippet: Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p
    Figure Legend Snippet: STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Real-time Polymerase Chain Reaction

    Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p
    Figure Legend Snippet: Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Enzyme-linked Immunosorbent Assay

    Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p
    Figure Legend Snippet: Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p

    Techniques Used: Infection, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p
    Figure Legend Snippet: Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    12) Product Images from "The formyl peptide receptor like-1 and scavenger receptor MARCO are involved in glial cell activation in bacterial meningitis"

    Article Title: The formyl peptide receptor like-1 and scavenger receptor MARCO are involved in glial cell activation in bacterial meningitis

    Journal: Journal of Neuroinflammation

    doi: 10.1186/1742-2094-8-11

    Inhibition of Neisseria meningitidis - and Streptococcus pneumoniae- induced ERK1/2 phosphorylation, and changes of cAMP levels by WRW4 and PTX in glial cells . For analysis of ERK1/2 phosphorylation, astrocytes (A) and microglia (B) were each treated with Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP); with NM as well as SP for 5 min at 37°C with 200 ng/ml PTX (16 h preincubation) or 10 μM WRW4 (30 min preincubation); they were also treated with PTX (16 h preincubation) or WRW4 (30 min preincubation) alone as control. Cells were lysed, equal amounts of protein (5 μg) were dissolved in SDS sample buffer, and the levels of total ERK2 and phosphorylated ERK1/2 were determined via immunoblotting. The positions of phospho-ERK1/2 (pERK1/2) and total ERK2 (ERK2) along with those of the molecular mass markers (in kDa) are indicated on the right or left side, respectively. The values representing mean ± standard error of the mean (SEM) of phosphorylation levels derived from densitometric quantification of three independent experiments in astrocytes and microglia are indicated in (C) and (D) , respectively. An asterisk indicates a significant difference (*, p
    Figure Legend Snippet: Inhibition of Neisseria meningitidis - and Streptococcus pneumoniae- induced ERK1/2 phosphorylation, and changes of cAMP levels by WRW4 and PTX in glial cells . For analysis of ERK1/2 phosphorylation, astrocytes (A) and microglia (B) were each treated with Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP); with NM as well as SP for 5 min at 37°C with 200 ng/ml PTX (16 h preincubation) or 10 μM WRW4 (30 min preincubation); they were also treated with PTX (16 h preincubation) or WRW4 (30 min preincubation) alone as control. Cells were lysed, equal amounts of protein (5 μg) were dissolved in SDS sample buffer, and the levels of total ERK2 and phosphorylated ERK1/2 were determined via immunoblotting. The positions of phospho-ERK1/2 (pERK1/2) and total ERK2 (ERK2) along with those of the molecular mass markers (in kDa) are indicated on the right or left side, respectively. The values representing mean ± standard error of the mean (SEM) of phosphorylation levels derived from densitometric quantification of three independent experiments in astrocytes and microglia are indicated in (C) and (D) , respectively. An asterisk indicates a significant difference (*, p

    Techniques Used: Inhibition, Derivative Assay

    Increased CAMP (human) expression in transfected HEK293 cells . For analysis of CAMP mRNA expression, untransfected (A) or hMARCO- (B) , hFPRL1- (C) , and hFPRL1/hMARCO- (D) expressing HEK293 cells were treated with Neisseria meningitidis (NM), Streptococcus pneumoniae (SP) or 1 μM fMLF for 0, 6, 12 and 24 h. mRNA expression was analyzed using TaqMan real-time RT-PCR and results were compared to the untreated sample. 18 s (housekeeping gene) was used as an internal control. The data was assessed from three independent experiments.
    Figure Legend Snippet: Increased CAMP (human) expression in transfected HEK293 cells . For analysis of CAMP mRNA expression, untransfected (A) or hMARCO- (B) , hFPRL1- (C) , and hFPRL1/hMARCO- (D) expressing HEK293 cells were treated with Neisseria meningitidis (NM), Streptococcus pneumoniae (SP) or 1 μM fMLF for 0, 6, 12 and 24 h. mRNA expression was analyzed using TaqMan real-time RT-PCR and results were compared to the untreated sample. 18 s (housekeeping gene) was used as an internal control. The data was assessed from three independent experiments.

    Techniques Used: Expressing, Transfection, Quantitative RT-PCR

    FPRL1- and MARCO-mediated ERK1/2 phosphorylation and changes of cAMP levels in transfected HEK293 cells . For analysis of ERK1/2 phosphorylation, untransfected (A) or hMARCO- (B) , hFPRL1- (C) , and hFPRL1/hMARCO- (D) expressing HEK293 cells were treated with Neisseria meningitidis (NM), Streptococcus pneumoniae (SP) or 1 μM fMLF for 5 and 15 min at 37°C. Cells were lysed, equal amounts of protein (5 μg) were dissolved by SDS sample buffer, and levels of total ERK2 and phosphorylated ERK1/2 were determined via immunoblotting. The positions of molecular mass markers are indicated on the left (in kDa). The mean ± SD of the three independent experiments were evaluated by densitometric quantification normalized to ERK2 expression (E to H) . Asterisks indicate a significant difference (*, p
    Figure Legend Snippet: FPRL1- and MARCO-mediated ERK1/2 phosphorylation and changes of cAMP levels in transfected HEK293 cells . For analysis of ERK1/2 phosphorylation, untransfected (A) or hMARCO- (B) , hFPRL1- (C) , and hFPRL1/hMARCO- (D) expressing HEK293 cells were treated with Neisseria meningitidis (NM), Streptococcus pneumoniae (SP) or 1 μM fMLF for 5 and 15 min at 37°C. Cells were lysed, equal amounts of protein (5 μg) were dissolved by SDS sample buffer, and levels of total ERK2 and phosphorylated ERK1/2 were determined via immunoblotting. The positions of molecular mass markers are indicated on the left (in kDa). The mean ± SD of the three independent experiments were evaluated by densitometric quantification normalized to ERK2 expression (E to H) . Asterisks indicate a significant difference (*, p

    Techniques Used: Transfection, Expressing

    Strongly increased MARCO expression in astrocytes after Neisseria meningitides infection in an infant rat model of meningitis . Coronal brain sections from rats infected with Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP) were fixed at defined time-points after infection and immunolabeled using anti-MARCO (green) and anti-glial fibrillary acidic protein (anti-GFAP) antibodies to identify astrocytes with nuclear counterstaining (blue). Sections were examined by double fluorescence microscopy. The figures show representative results from one of three independent experiments. Scale bar = 200 μm for overview and 20 μm for the other images. (B) Shows a detail of (A) . Please note that the detail shows a strong colocalization between GFAP and MARCO in the sub-cortical layer after NM infection.
    Figure Legend Snippet: Strongly increased MARCO expression in astrocytes after Neisseria meningitides infection in an infant rat model of meningitis . Coronal brain sections from rats infected with Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP) were fixed at defined time-points after infection and immunolabeled using anti-MARCO (green) and anti-glial fibrillary acidic protein (anti-GFAP) antibodies to identify astrocytes with nuclear counterstaining (blue). Sections were examined by double fluorescence microscopy. The figures show representative results from one of three independent experiments. Scale bar = 200 μm for overview and 20 μm for the other images. (B) Shows a detail of (A) . Please note that the detail shows a strong colocalization between GFAP and MARCO in the sub-cortical layer after NM infection.

    Techniques Used: Expressing, Infection, Immunolabeling, Fluorescence, Microscopy

    Inhibition of Neisseria meningitidis as well as Streptococcus pneumoniae -induced Camp (rat) expression by the FPRL1 antagonist WRW4 and the G-protein inhibitor pertussis toxin in glial cells . Bacterial supernatants from Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP) were added to astrocytes (A) and microglia (B) with the addition of 200 ng/ml PTX (16 h preincubation) or 10 μM WRW4 (30 min preincubation) and with PTX (16 h preincubation) and WRW4 (30 min preincubation) alone to analyze the effect on Camp mRNA expression after 24 h (astrocytes; a ) or 6 h (microglia; b ) of treatment. The induction was analyzed and compared to an untreated sample (also with DMSO in equivalent amount). GAPDH (housekeeping gene) was used as an internal control. The data from three independent experiments performed in triplicate were assessed. An asterisk (*, p
    Figure Legend Snippet: Inhibition of Neisseria meningitidis as well as Streptococcus pneumoniae -induced Camp (rat) expression by the FPRL1 antagonist WRW4 and the G-protein inhibitor pertussis toxin in glial cells . Bacterial supernatants from Neisseria meningitidis (NM) or Streptococcus pneumoniae (SP) were added to astrocytes (A) and microglia (B) with the addition of 200 ng/ml PTX (16 h preincubation) or 10 μM WRW4 (30 min preincubation) and with PTX (16 h preincubation) and WRW4 (30 min preincubation) alone to analyze the effect on Camp mRNA expression after 24 h (astrocytes; a ) or 6 h (microglia; b ) of treatment. The induction was analyzed and compared to an untreated sample (also with DMSO in equivalent amount). GAPDH (housekeeping gene) was used as an internal control. The data from three independent experiments performed in triplicate were assessed. An asterisk (*, p

    Techniques Used: Inhibition, Expressing

    Bacterial supernatants of Neisseria meningitidis induced MARCO expression in primary rat astrocytes and meningeal cells . Astrocytes (A and B), microglia (C and D) and meningeal cells (E and F) were incubated with bacterial supernatants of Gram-positive bacteria Streptococcus pneumoniae (SP) or Gram-negative bacteria Neisseria meningitidis (NM) for 0, 6, 12 and 24 h. FPRL1 or MARCO mRNA expression was analyzed using SYBR green real-time RT-PCR and results were compared to the untreated sample. GAPDH (housekeeping gene) was used as an internal control. The data were assessed from three independent experiments in triplicate. An asterisk indicates a significant difference (**, p
    Figure Legend Snippet: Bacterial supernatants of Neisseria meningitidis induced MARCO expression in primary rat astrocytes and meningeal cells . Astrocytes (A and B), microglia (C and D) and meningeal cells (E and F) were incubated with bacterial supernatants of Gram-positive bacteria Streptococcus pneumoniae (SP) or Gram-negative bacteria Neisseria meningitidis (NM) for 0, 6, 12 and 24 h. FPRL1 or MARCO mRNA expression was analyzed using SYBR green real-time RT-PCR and results were compared to the untreated sample. GAPDH (housekeeping gene) was used as an internal control. The data were assessed from three independent experiments in triplicate. An asterisk indicates a significant difference (**, p

    Techniques Used: Expressing, Incubation, SYBR Green Assay, Quantitative RT-PCR

    13) Product Images from "NLRP3 inflammasome activation in aged macrophages is diminished during Streptococcus pneumoniae infection"

    Article Title: NLRP3 inflammasome activation in aged macrophages is diminished during Streptococcus pneumoniae infection

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00393.2017

    Similar NF-κB expression in young and aged macrophages in response to S. pneumoniae . A – D : young and aged macrophages were stimulated with TLR-1/2 (Pam3CSK4, 100 ng/ml), TLR-2 (HKLM, 10 7 cells/ml), TLR-3 (HMW poly I:C, 10 ng/ml), TLR-3 (LMW poly I:C, 30 ng/ml), TLR-4 (LPS-EK, 10 ng/ml), TLR-5 (FLA-st, 100 ng/ml), TLR-6 (FSL-1 10 ng/ml), TLR-7 (ssRNA40, 1 μg/ml), or TLR-9 (ODN1826, 1 μM) agonists. TNF-α (TLR-1/2: t -test, P = 0.0446; TLR-3 (HMW): t -test, * P = 0.0114; TLR-3 (LMW): t -test, *** P = 0.0006; TLR-9: t -test, * P = 0.0462) ( A ) and IL-1β (TLR-1/2: t -test, ** P = 0.0037; TLR-2: t -test, ** P = 0.0016; TLR-4: t -test, * P = 0.0155; TLR-6: t -test, ** P = 0.0097; TLR-9: t -test, *** P = 0.0002) ( B ) production was quantified by ELISA. C and D : cytosolic and nuclear protein extracts were isolated from young and aged macrophages at 2 h post- S. pneumoniae (50 CFU) infection. NF-κB ( t -test, **** P
    Figure Legend Snippet: Similar NF-κB expression in young and aged macrophages in response to S. pneumoniae . A – D : young and aged macrophages were stimulated with TLR-1/2 (Pam3CSK4, 100 ng/ml), TLR-2 (HKLM, 10 7 cells/ml), TLR-3 (HMW poly I:C, 10 ng/ml), TLR-3 (LMW poly I:C, 30 ng/ml), TLR-4 (LPS-EK, 10 ng/ml), TLR-5 (FLA-st, 100 ng/ml), TLR-6 (FSL-1 10 ng/ml), TLR-7 (ssRNA40, 1 μg/ml), or TLR-9 (ODN1826, 1 μM) agonists. TNF-α (TLR-1/2: t -test, P = 0.0446; TLR-3 (HMW): t -test, * P = 0.0114; TLR-3 (LMW): t -test, *** P = 0.0006; TLR-9: t -test, * P = 0.0462) ( A ) and IL-1β (TLR-1/2: t -test, ** P = 0.0037; TLR-2: t -test, ** P = 0.0016; TLR-4: t -test, * P = 0.0155; TLR-6: t -test, ** P = 0.0097; TLR-9: t -test, *** P = 0.0002) ( B ) production was quantified by ELISA. C and D : cytosolic and nuclear protein extracts were isolated from young and aged macrophages at 2 h post- S. pneumoniae (50 CFU) infection. NF-κB ( t -test, **** P

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Isolation, Infection

    Endoplasmic reticulum (ER) stress in the aged lung is enhanced during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in BIP/GRP78 expression in lung homogenates was calculated at baseline and at 24, 48, and 72 h postinfection (24 h: t -test, * P = 0.0189; 48 h: t -test, ** P = 0.0028; 72 h, t -test: ** P = 0.0020). B and C : inositol-requiring enzyme 1 (IRE1) and activating transcription factor 6 (ATF6) ( B ) and PKR-like ER resident kinase (PERK) ( C ) mRNA expression was quantified by real-time PCR at 24 h postinfection (IRE1: t -test, * P = 0.0119; PERK: t -test, ** P
    Figure Legend Snippet: Endoplasmic reticulum (ER) stress in the aged lung is enhanced during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in BIP/GRP78 expression in lung homogenates was calculated at baseline and at 24, 48, and 72 h postinfection (24 h: t -test, * P = 0.0189; 48 h: t -test, ** P = 0.0028; 72 h, t -test: ** P = 0.0020). B and C : inositol-requiring enzyme 1 (IRE1) and activating transcription factor 6 (ATF6) ( B ) and PKR-like ER resident kinase (PERK) ( C ) mRNA expression was quantified by real-time PCR at 24 h postinfection (IRE1: t -test, * P = 0.0119; PERK: t -test, ** P

    Techniques Used: Infection, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    AIM2 inflammasome responses are not essential for caspase-1 activation or IL-1β production in response to a serotype 3 strain of S. pneumoniae . Young wild-type (WT) and Aim2 −/− macrophages were cultured with S. pneumoniae ( S. pne ) (50 CFU) and mRNA and protein expression was examined at 4 and 24 h postinfection. A : changes in TLR- 1, 2, 4, 6, and 9, and MYD88 were quantified by real-time PCR. B : mRNA expression of NOD1, NOD2 (4 h: t -test, P
    Figure Legend Snippet: AIM2 inflammasome responses are not essential for caspase-1 activation or IL-1β production in response to a serotype 3 strain of S. pneumoniae . Young wild-type (WT) and Aim2 −/− macrophages were cultured with S. pneumoniae ( S. pne ) (50 CFU) and mRNA and protein expression was examined at 4 and 24 h postinfection. A : changes in TLR- 1, 2, 4, 6, and 9, and MYD88 were quantified by real-time PCR. B : mRNA expression of NOD1, NOD2 (4 h: t -test, P

    Techniques Used: Activation Assay, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Treatment with TUDCA rescues NLRP3 inflammasome activation in the aged lung during S. pneumoniae infection. A – E ; young (2 mo) and aged (18 mo) male and female BALB/c mice received saline or TUDCA (500 mg·kg −1 ·day −1 ) via intraperitoneal injection before intranasal instillation with saline or 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : gene expression in lung tissue collected from young, aged, and TUDCA-treated aged mice at 24 h postinfection was quantified by real-time PCR using inflammasome specific primer assays (SABiosciences). ASC, NLRP3, and pro-caspase-1 mRNA expression was evaluated by real-time PCR (ASC: t -test, **** P
    Figure Legend Snippet: Treatment with TUDCA rescues NLRP3 inflammasome activation in the aged lung during S. pneumoniae infection. A – E ; young (2 mo) and aged (18 mo) male and female BALB/c mice received saline or TUDCA (500 mg·kg −1 ·day −1 ) via intraperitoneal injection before intranasal instillation with saline or 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : gene expression in lung tissue collected from young, aged, and TUDCA-treated aged mice at 24 h postinfection was quantified by real-time PCR using inflammasome specific primer assays (SABiosciences). ASC, NLRP3, and pro-caspase-1 mRNA expression was evaluated by real-time PCR (ASC: t -test, **** P

    Techniques Used: Activation Assay, Infection, Mouse Assay, Injection, Expressing, Real-time Polymerase Chain Reaction

    Macrophage and lung Toll-like receptor (TLR) expression in response to S. pneumoniae . A – C : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in mRNA expression of TLR-1 ( t -test, * P = 0.01354), 2 ( t -test, ** P = 0.0082), 4 ( t -test, * P = 0.0487), 6, and 9 ( t -test, * P = 0.0144) in young and aged lungs was evaluated by real-time PCR. B : changes in mRNA expression of NOD1 ( t -test, * P = 0.0054), NOD2 ( t -test, ** P = 0.015), RIP2, and NF-κB at 24 h postinfection were quantified by real-time PCR. C : inflammatory cytokine expression was investigated in young and aged lungs, and changes in pro-IL-1β, pro-IL-18, IL-6 ( t -test, **** P
    Figure Legend Snippet: Macrophage and lung Toll-like receptor (TLR) expression in response to S. pneumoniae . A – C : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in mRNA expression of TLR-1 ( t -test, * P = 0.01354), 2 ( t -test, ** P = 0.0082), 4 ( t -test, * P = 0.0487), 6, and 9 ( t -test, * P = 0.0144) in young and aged lungs was evaluated by real-time PCR. B : changes in mRNA expression of NOD1 ( t -test, * P = 0.0054), NOD2 ( t -test, ** P = 0.015), RIP2, and NF-κB at 24 h postinfection were quantified by real-time PCR. C : inflammatory cytokine expression was investigated in young and aged lungs, and changes in pro-IL-1β, pro-IL-18, IL-6 ( t -test, **** P

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Treatment with tauroursodeoxycholic acid improves NLRP3 inflammasome activation in aged macrophages during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male or female BALB/c macrophages were pretreated with tauroursodeoxycholic acid (TUDCA) (100 μM) for 24 h before culture with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : mRNA expression of ATF6, IRE1, and PERK in untreated and TUDCA-treated aged macrophages at 24 h postinfection was quantified by real-time PCR (IRE1: t -test, *** P = 0.0008). B : IRE1 phosphorylation in untreated and TUDCA pretreated aged macrophages at 24 h postinfection was examined by Western blot. C : XBP1 splicing was evaluated by PCR and % of XBP1 splicing was quantified ( t -test, ** P = 0.0032). D : cytotoxicity in young and aged macrophages in response to TUDCA was examined at 24 h post- S. pneumoniae infection ( t -test, ** P
    Figure Legend Snippet: Treatment with tauroursodeoxycholic acid improves NLRP3 inflammasome activation in aged macrophages during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male or female BALB/c macrophages were pretreated with tauroursodeoxycholic acid (TUDCA) (100 μM) for 24 h before culture with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : mRNA expression of ATF6, IRE1, and PERK in untreated and TUDCA-treated aged macrophages at 24 h postinfection was quantified by real-time PCR (IRE1: t -test, *** P = 0.0008). B : IRE1 phosphorylation in untreated and TUDCA pretreated aged macrophages at 24 h postinfection was examined by Western blot. C : XBP1 splicing was evaluated by PCR and % of XBP1 splicing was quantified ( t -test, ** P = 0.0032). D : cytotoxicity in young and aged macrophages in response to TUDCA was examined at 24 h post- S. pneumoniae infection ( t -test, ** P

    Techniques Used: Activation Assay, Infection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Polymerase Chain Reaction

    NLRP3 expression in the aged lung is necessary for survival during Streptococcus pneumoniae . A , D – H : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 colony-forming untis (CFU) of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : hematoxylin and eosin (H E) staining of lung tissue from young and aged control and S. pneumoniae -infected mice collected at 72 h postinfection. B and C : young (2 mo) and aged (17 mo) male and female wild-type C57BL/6 or C57Bl/6-NLRP3 −/− mice were instilled with 1 × 10 3 CFU of S. pneumoniae . Survival ( B ) (Mantel-Cox: P = 0.0026) and bacterial titer ( C ) in lung was examined in lung homogenates at 24 h postinfection ( t -test, ** P
    Figure Legend Snippet: NLRP3 expression in the aged lung is necessary for survival during Streptococcus pneumoniae . A , D – H : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 colony-forming untis (CFU) of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : hematoxylin and eosin (H E) staining of lung tissue from young and aged control and S. pneumoniae -infected mice collected at 72 h postinfection. B and C : young (2 mo) and aged (17 mo) male and female wild-type C57BL/6 or C57Bl/6-NLRP3 −/− mice were instilled with 1 × 10 3 CFU of S. pneumoniae . Survival ( B ) (Mantel-Cox: P = 0.0026) and bacterial titer ( C ) in lung was examined in lung homogenates at 24 h postinfection ( t -test, ** P

    Techniques Used: Expressing, Mouse Assay, Staining, Infection

    Diminished NLRP3 inflammasome activation in aged macrophages in response to S. pneumoniae . Young (2 mo) and aged (18 mo) male and female BALB/c macrophages were treated with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ). A : baseline and S. pneumoniae upregulated pro-caspase-1, NLRP3, and ASC mRNA expression was evaluated by real-time PCR ( t -test, *** P
    Figure Legend Snippet: Diminished NLRP3 inflammasome activation in aged macrophages in response to S. pneumoniae . Young (2 mo) and aged (18 mo) male and female BALB/c macrophages were treated with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ). A : baseline and S. pneumoniae upregulated pro-caspase-1, NLRP3, and ASC mRNA expression was evaluated by real-time PCR ( t -test, *** P

    Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction

    14) Product Images from "Roles of STAT3 in Protein Secretion Pathways during the Acute-Phase Response"

    Article Title: Roles of STAT3 in Protein Secretion Pathways during the Acute-Phase Response

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01332-12

    Secretory protein gene induction is diminished by simultaneous mutation of both STAT3 and RelA. Stat3/RelAhepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA
    Figure Legend Snippet: Secretory protein gene induction is diminished by simultaneous mutation of both STAT3 and RelA. Stat3/RelAhepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA

    Techniques Used: Mutagenesis, Mouse Assay

    Expression of UPR target genes is diminished by STAT3 mutation. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis
    Figure Legend Snippet: Expression of UPR target genes is diminished by STAT3 mutation. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, RNA Extraction, Quantitative RT-PCR

    Secretory protein gene induction requires STAT3. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis of secretory
    Figure Legend Snippet: Secretory protein gene induction requires STAT3. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis of secretory

    Techniques Used: Mouse Assay, RNA Extraction, Quantitative RT-PCR

    Secretory pathway gene induction during pneumococcal pneumonia. Quantitative RT-PCR was performed on liver RNA collected from B6/129 mice 0 to 24 h after intratracheal S. pneumoniae infection to evaluate transcripts involved in protein translation into
    Figure Legend Snippet: Secretory pathway gene induction during pneumococcal pneumonia. Quantitative RT-PCR was performed on liver RNA collected from B6/129 mice 0 to 24 h after intratracheal S. pneumoniae infection to evaluate transcripts involved in protein translation into

    Techniques Used: Quantitative RT-PCR, Mouse Assay, Infection

    Secretory protein gene induction is unaffected by mutation of RelA. RelAhepΔ mice and control littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis
    Figure Legend Snippet: Secretory protein gene induction is unaffected by mutation of RelA. RelAhepΔ mice and control littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis

    Techniques Used: Mutagenesis, Mouse Assay, RNA Extraction, Quantitative RT-PCR

    15) Product Images from "Depletion of Human DNA in Spiked Clinical Specimens for Improvement of Sensitivity of Pathogen Detection by Next-Generation Sequencing"

    Article Title: Depletion of Human DNA in Spiked Clinical Specimens for Improvement of Sensitivity of Pathogen Detection by Next-Generation Sequencing

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.03050-15

    Processing of clinical specimens using various detergents for selective lysis of human cells. NPA specimens were spiked with S. pneumoniae , HSV2, and adenovirus and treated with saponin, Triton X-100, Tween 20, CHAPS buffer, or MolZym CM buffer to final
    Figure Legend Snippet: Processing of clinical specimens using various detergents for selective lysis of human cells. NPA specimens were spiked with S. pneumoniae , HSV2, and adenovirus and treated with saponin, Triton X-100, Tween 20, CHAPS buffer, or MolZym CM buffer to final

    Techniques Used: Lysis

    Analysis of clinical specimens processed with 0.025% saponin and DNase by PCR and NGS. CSF specimens ( n = 3) spiked with S. pneumoniae , N. meningitidis , H. influenzae , E. coli , S. agalactiae , HSV2, and adenovirus and NPA specimens ( n = 3) spiked with
    Figure Legend Snippet: Analysis of clinical specimens processed with 0.025% saponin and DNase by PCR and NGS. CSF specimens ( n = 3) spiked with S. pneumoniae , N. meningitidis , H. influenzae , E. coli , S. agalactiae , HSV2, and adenovirus and NPA specimens ( n = 3) spiked with

    Techniques Used: Polymerase Chain Reaction, Next-Generation Sequencing

    Processing of clinical specimens using saponin at different concentrations for selective lysis of human cells. CSF specimens were spiked with S. pneumoniae , H. influenzae , E. coli , HSV2, and adenovirus (A) and NPA specimens spiked with S. pneumoniae ,
    Figure Legend Snippet: Processing of clinical specimens using saponin at different concentrations for selective lysis of human cells. CSF specimens were spiked with S. pneumoniae , H. influenzae , E. coli , HSV2, and adenovirus (A) and NPA specimens spiked with S. pneumoniae ,

    Techniques Used: Lysis

    Processing of clinical specimens using MolYsis reagents. CSF (A) or NPA (B) specimens were spiked with S. pneumoniae and influenza A virus and processed by MolYsis method I and MolYsis method II, as described in the Materials and Methods. Nucleic acids
    Figure Legend Snippet: Processing of clinical specimens using MolYsis reagents. CSF (A) or NPA (B) specimens were spiked with S. pneumoniae and influenza A virus and processed by MolYsis method I and MolYsis method II, as described in the Materials and Methods. Nucleic acids

    Techniques Used:

    16) Product Images from "Antimicrobial Properties of Mesenchymal Stem Cells: Therapeutic Potential for Cystic Fibrosis Infection, and Treatment"

    Article Title: Antimicrobial Properties of Mesenchymal Stem Cells: Therapeutic Potential for Cystic Fibrosis Infection, and Treatment

    Journal: Stem Cells International

    doi: 10.1155/2016/5303048

    hMSCs and their products on Streptococcus pneumoniae growth. Bone marrow derived hMSCs supernatants were cultured with different dosages of Streptococcus pneumoniae with and without the addition of geneticin (100 μ g/mL). Aliquots of the bacteria were streaked onto MacConkey plates for CFUs (a) or evaluated for ATP production (b). hMSC supernatants ( n = 8 different donors) significantly decreased both Streptococcus pneumoniae CFUs ((a), P ≤ 0.05) and growth rates ((b), P ≤ 0.05). Geneticin was used as a positive control, decreasing both CFUs ( P ≤ 0.05) and growth rates ( P ≤ 0.05). hMSC supernatants decreased CFUs and enhanced antibiotic sensitivity when measuring CFUs. However, hMSCs supernatants had minimal antibiotic enhancing effect on Streptococcus pneumonia growth rate (b). ST = Streptococcus pneumoniae growth without treatment. Geneticin = treatment with the antibiotic geneticin, +hMSCs = treatment with hMSC derived supernatant, and Both = treatment with both MSC supernatants and geneticin.
    Figure Legend Snippet: hMSCs and their products on Streptococcus pneumoniae growth. Bone marrow derived hMSCs supernatants were cultured with different dosages of Streptococcus pneumoniae with and without the addition of geneticin (100 μ g/mL). Aliquots of the bacteria were streaked onto MacConkey plates for CFUs (a) or evaluated for ATP production (b). hMSC supernatants ( n = 8 different donors) significantly decreased both Streptococcus pneumoniae CFUs ((a), P ≤ 0.05) and growth rates ((b), P ≤ 0.05). Geneticin was used as a positive control, decreasing both CFUs ( P ≤ 0.05) and growth rates ( P ≤ 0.05). hMSC supernatants decreased CFUs and enhanced antibiotic sensitivity when measuring CFUs. However, hMSCs supernatants had minimal antibiotic enhancing effect on Streptococcus pneumonia growth rate (b). ST = Streptococcus pneumoniae growth without treatment. Geneticin = treatment with the antibiotic geneticin, +hMSCs = treatment with hMSC derived supernatant, and Both = treatment with both MSC supernatants and geneticin.

    Techniques Used: Derivative Assay, Cell Culture, Positive Control

    17) Product Images from "Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection"

    Article Title: Age Enhanced ER Stress Contributes to Increased Atg9A Inhibition of STING Mediated IFNβ Production during Streptococcus Pneumoniae Infection

    Journal: Journal of immunology (Baltimore, Md. : 1950)

    doi: 10.4049/jimmunol.1303090

    Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p
    Figure Legend Snippet: Increased Atg9A Mediated Inhibition of STING in Aged Lung during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone or with S. pne for 24 hours. Protein was isolated and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (B) Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne or saline via intranasal instillation. Protein was collected from lung tissue samples at 24 hours post infection and expression of IRE1, XBP1, and Atg9a were assessed by western blotting. (C) Co-immunoprecipitation using α-Atg9a followed by western blotting for acetylated lysine (one-way ANOVA, p=0.0391). (D-F) Aged macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. (D) Protein was isolated and expression of BIP/GRP78, IRE1, XBP1, and Atg9a in aged macrophages was assessed by western blot. (E) STING protein and IRF3 activation were assessed by ELISA (t-test comparisons of media and compound treatment, p

    Techniques Used: Inhibition, Infection, Cell Culture, Isolation, Expressing, Western Blot, Mouse Assay, Immunoprecipitation, Activation Assay, Enzyme-linked Immunosorbent Assay, T-Test

    Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p
    Figure Legend Snippet: Gemcitabine HCl Decreases Atg9A Mediated Inhibition of STING and Enhances IFNβ Production during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. Starting at 4 hours post infection, animals received a 100μL intraperitoneal injection of gemcitabine HCl (0.3mg/mL) or saline control. (A-C) Weights (A: two-way ANOVA, p

    Techniques Used: Inhibition, Infection, Mouse Assay, Injection

    Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p
    Figure Legend Snippet: Decreased IFNβ Production in Aged Mice during S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. IFNβ production in (A) lung homogenates (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p
    Figure Legend Snippet: STING Mediated Production of IFNβ is Decreased in Aged Hosts during S. pneumoniae infection (A-C) Macrophages were cultured with media alone or with S. pne (50 CFU) for 24 hours prior to RNA or protein isolation. (A) Gene specific expression of STING, TBK1, IRF3, and IFNβ, relative to media control, was assessed by real time PCR (two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Real-time Polymerase Chain Reaction

    Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p
    Figure Legend Snippet: Enhanced ER Stress in Aged Hosts during S. pneumoniae Infection (A-B) Macrophages were cultured with media alone, media containing tauroursodeoxycholic acid (TUDCA), sunitinib malate (SM), and/or S. pne overnight. Protein was isolated and expression of BIP/GRP78 and PDI were assessed by ELISA (A: two-way ANOVA, p

    Techniques Used: Infection, Cell Culture, Isolation, Expressing, Enzyme-linked Immunosorbent Assay

    Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p
    Figure Legend Snippet: Knockdown of Atg9a Restores STING Mediated Production of IFNβ Production during S. pneumoniae Infection (A-G) Cells were treated with missense or Atg9a specific siRNA for 24 hours prior to S. pne infection. (A) RNA was isolated from cultured macrophages and expression, relative to control was assessed for Atg9A, STING, IRF3, and TBK1 by real time PCR. Gene expression was normalized to β2M (A: two-way ANOVA, p

    Techniques Used: Infection, Isolation, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p
    Figure Legend Snippet: Increased Morbidity in Aged Mice in Response to S. pneumoniae Infection Young (2 months) and aged (19 months) male and female BALB/c mice received 1×10 3 CFU of S. pne (ATCC 6303) or saline via intranasal instillation. (A) Weights were collected at 24 and 48 hours post infection (two-way ANOVA, p

    Techniques Used: Mouse Assay, Infection

    18) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    19) Product Images from "Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H"

    Article Title: Resistance to Both Complement Activation and Phagocytosis in Type 3 Pneumococci Is Mediated by the Binding of Complement Regulatory Protein Factor H

    Journal: Infection and Immunity

    doi:

    Survival of mice after intraperitoneal challenge with S. pneumoniae ATCC 6303 (Caps + /PspA + ) pretreated with trypsin or buffer as a control (see Materials and Methods). The curves show survival time (days) of mice after an i.p. injection of 20 CFU/mouse (15 mice per group). Trypsin-treated pneumococci were significantly different ( P
    Figure Legend Snippet: Survival of mice after intraperitoneal challenge with S. pneumoniae ATCC 6303 (Caps + /PspA + ) pretreated with trypsin or buffer as a control (see Materials and Methods). The curves show survival time (days) of mice after an i.p. injection of 20 CFU/mouse (15 mice per group). Trypsin-treated pneumococci were significantly different ( P

    Techniques Used: Mouse Assay, Injection

    20) Product Images from "Reduced inflammation and cytokine production in NKLAM deficient mice during Streptococcus pneumoniae infection"

    Article Title: Reduced inflammation and cytokine production in NKLAM deficient mice during Streptococcus pneumoniae infection

    Journal: PLoS ONE

    doi: 10.1371/journal.pone.0194202

    S . pneumoniae high-dose survival experiment. WT and NKLAM-KO mice were infected via intranasal route with 5 x 10 7 S . pneumoniae CFU in 30 μL sterile PBS and monitored for signs of severe illness for 5 days. WT, n = 11; NKLAM-KO, n = 12. p = not significant.
    Figure Legend Snippet: S . pneumoniae high-dose survival experiment. WT and NKLAM-KO mice were infected via intranasal route with 5 x 10 7 S . pneumoniae CFU in 30 μL sterile PBS and monitored for signs of severe illness for 5 days. WT, n = 11; NKLAM-KO, n = 12. p = not significant.

    Techniques Used: Mouse Assay, Infection

    Lack of NKLAM is associated with reduced lung inflammation. (A) Lungs from 24h-infected or (B) 48h-infected S . pneumoniae -infected (10 x 10 6 CFU/mouse) WT and NKLAM-KO or PBS-treated mice were formalin-fixed, embedded in paraffin and sections were stained with H E or anti-mouse MPO. Magnification, x 200 for H E, x 400 for MPO. Black bar equals 50 μm. Data are representative of two experiments, with n = 2 mice per group per experiment.
    Figure Legend Snippet: Lack of NKLAM is associated with reduced lung inflammation. (A) Lungs from 24h-infected or (B) 48h-infected S . pneumoniae -infected (10 x 10 6 CFU/mouse) WT and NKLAM-KO or PBS-treated mice were formalin-fixed, embedded in paraffin and sections were stained with H E or anti-mouse MPO. Magnification, x 200 for H E, x 400 for MPO. Black bar equals 50 μm. Data are representative of two experiments, with n = 2 mice per group per experiment.

    Techniques Used: Infection, Mouse Assay, Staining

    The absence of NKLAM impairs S . pneumoniae killing. (A) WT and NKLAM-KO mice were infected by nasal administration of 10 x 10 6 CFU of S . pneumoniae . The lungs were harvested at 24 or 48h, homogenized in sterile PBS and an aliquot was plated on blood agar plates to determine bacterial load. Instillation of sterile PBS was used as a control. Data represent mean ± SD of 1 of 3 independent experiments. (n = 8–11 total mice for each condition); * p
    Figure Legend Snippet: The absence of NKLAM impairs S . pneumoniae killing. (A) WT and NKLAM-KO mice were infected by nasal administration of 10 x 10 6 CFU of S . pneumoniae . The lungs were harvested at 24 or 48h, homogenized in sterile PBS and an aliquot was plated on blood agar plates to determine bacterial load. Instillation of sterile PBS was used as a control. Data represent mean ± SD of 1 of 3 independent experiments. (n = 8–11 total mice for each condition); * p

    Techniques Used: Mouse Assay, Infection

    Neutrophil and NK cell lung infiltration and bone marrow-derived macrophage iNOS expression. Cells were isolated from infected lungs at 24 and 48h post infection and stained for CD45, CD3, NK1.1, CD11b and Ly-6G. CD45 + cells were gated and the percentage of CD11b + /Ly-6G + (A) or CD3 - /NK1.1 + (B) cells within the CD45 + population was determined. A representative histogram is shown for each group (n = 3–4 mice per group). (C) BMDM were treated with 100 μg/mL lipoteichoic acid (LTA) or formalin-fixed S . pneumoniae (SP) at an MOI of 10 for 18 hr at 37°C and protein lysates were immunoblotted for iNOS protein. Beta actin was used as a loading control. Immunoblots represent 1 of 3 identical experiments, (D) Graphical representation of C. n = 3; *p
    Figure Legend Snippet: Neutrophil and NK cell lung infiltration and bone marrow-derived macrophage iNOS expression. Cells were isolated from infected lungs at 24 and 48h post infection and stained for CD45, CD3, NK1.1, CD11b and Ly-6G. CD45 + cells were gated and the percentage of CD11b + /Ly-6G + (A) or CD3 - /NK1.1 + (B) cells within the CD45 + population was determined. A representative histogram is shown for each group (n = 3–4 mice per group). (C) BMDM were treated with 100 μg/mL lipoteichoic acid (LTA) or formalin-fixed S . pneumoniae (SP) at an MOI of 10 for 18 hr at 37°C and protein lysates were immunoblotted for iNOS protein. Beta actin was used as a loading control. Immunoblots represent 1 of 3 identical experiments, (D) Graphical representation of C. n = 3; *p

    Techniques Used: Derivative Assay, Expressing, Isolation, Infection, Staining, Mouse Assay, Western Blot

    S . pneumoniae -induced phosphorylation of STAT1 and STAT3 is diminished in NKLAM-KO mice. Lung homogenates from S . pneumoniae -infected WT and NKLAM-KO mice were immunoblotted for (A) STAT1 and pSTAT1 (Tyr701) and (B) STAT3 and pSTAT3 (Tyr705). Each lane represents an individual mouse and is representative of the overall experiments. Immunoblots were used to calculate the ratio of pSTAT1 (Tyr701)/STAT1 (C) and pSTAT3 (Tyr705)/STAT3 (D). Data represent the mean ± SD of 1 of 3 representative experiments (n = 3–5 mice per group per experiment); * p ≤ 0.05. (E) Sections from paraffin-embedded lungs from S . pneumoniae -infected (24h) WT and NKLAM-KO mice were stained for pSTAT1 (Tyr701) (red) and DAPI (blue). Colocalization is depicted in pink (arrows). (n = 2–3 mice per group). (F) Aliquots (50 μg) of lung homogenate from PBS control mice and S . pneumoniae -infected (24h) WT and NKLAM-KO mice were separated by native gel electrophoresis then incubated with 4-methylumbelliferyl phosphate to visualize phosphatase activity. Each lane represents an individual mouse. Graph represents the mean band intensity ± SD. Results are representative of two experiments with 3–5 mice per group per experiment; * p ≤ 0.05.
    Figure Legend Snippet: S . pneumoniae -induced phosphorylation of STAT1 and STAT3 is diminished in NKLAM-KO mice. Lung homogenates from S . pneumoniae -infected WT and NKLAM-KO mice were immunoblotted for (A) STAT1 and pSTAT1 (Tyr701) and (B) STAT3 and pSTAT3 (Tyr705). Each lane represents an individual mouse and is representative of the overall experiments. Immunoblots were used to calculate the ratio of pSTAT1 (Tyr701)/STAT1 (C) and pSTAT3 (Tyr705)/STAT3 (D). Data represent the mean ± SD of 1 of 3 representative experiments (n = 3–5 mice per group per experiment); * p ≤ 0.05. (E) Sections from paraffin-embedded lungs from S . pneumoniae -infected (24h) WT and NKLAM-KO mice were stained for pSTAT1 (Tyr701) (red) and DAPI (blue). Colocalization is depicted in pink (arrows). (n = 2–3 mice per group). (F) Aliquots (50 μg) of lung homogenate from PBS control mice and S . pneumoniae -infected (24h) WT and NKLAM-KO mice were separated by native gel electrophoresis then incubated with 4-methylumbelliferyl phosphate to visualize phosphatase activity. Each lane represents an individual mouse. Graph represents the mean band intensity ± SD. Results are representative of two experiments with 3–5 mice per group per experiment; * p ≤ 0.05.

    Techniques Used: Mouse Assay, Infection, Western Blot, Staining, Nucleic Acid Electrophoresis, Incubation, Activity Assay

    Mouse inflammatory cytokine profile from S . pneumoniae -infected WT and NKLAM-KO mice. Mouse inflammatory cytokines (MCP-1, TNΦα, IFNγ and IL-12p70) were evaluated in lung homogenates (A) and plasma (B) from infected (24 and 48hr) WT and NKLAM-KO mice by cytometric bead array. Data shown were pooled from 3 independent experiments (n = 8–11 mice per group). * p ≤ 0.05.
    Figure Legend Snippet: Mouse inflammatory cytokine profile from S . pneumoniae -infected WT and NKLAM-KO mice. Mouse inflammatory cytokines (MCP-1, TNΦα, IFNγ and IL-12p70) were evaluated in lung homogenates (A) and plasma (B) from infected (24 and 48hr) WT and NKLAM-KO mice by cytometric bead array. Data shown were pooled from 3 independent experiments (n = 8–11 mice per group). * p ≤ 0.05.

    Techniques Used: Infection, Mouse Assay

    21) Product Images from "Roles of STAT3 in Protein Secretion Pathways during the Acute-Phase Response"

    Article Title: Roles of STAT3 in Protein Secretion Pathways during the Acute-Phase Response

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01332-12

    Secretory protein gene induction is diminished by simultaneous mutation of both STAT3 and RelA. Stat3/RelAhepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA
    Figure Legend Snippet: Secretory protein gene induction is diminished by simultaneous mutation of both STAT3 and RelA. Stat3/RelAhepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA

    Techniques Used: Mutagenesis, Mouse Assay

    Expression of UPR target genes is diminished by STAT3 mutation. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis
    Figure Legend Snippet: Expression of UPR target genes is diminished by STAT3 mutation. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis

    Techniques Used: Expressing, Mutagenesis, Mouse Assay, RNA Extraction, Quantitative RT-PCR

    Secretory protein gene induction requires STAT3. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis of secretory
    Figure Legend Snippet: Secretory protein gene induction requires STAT3. Stat3hepΔ mice and control (WT) littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis of secretory

    Techniques Used: Mouse Assay, RNA Extraction, Quantitative RT-PCR

    Secretory pathway gene induction during pneumococcal pneumonia. Quantitative RT-PCR was performed on liver RNA collected from B6/129 mice 0 to 24 h after intratracheal S. pneumoniae infection to evaluate transcripts involved in protein translation into
    Figure Legend Snippet: Secretory pathway gene induction during pneumococcal pneumonia. Quantitative RT-PCR was performed on liver RNA collected from B6/129 mice 0 to 24 h after intratracheal S. pneumoniae infection to evaluate transcripts involved in protein translation into

    Techniques Used: Quantitative RT-PCR, Mouse Assay, Infection

    Secretory protein gene induction is unaffected by mutation of RelA. RelAhepΔ mice and control littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis
    Figure Legend Snippet: Secretory protein gene induction is unaffected by mutation of RelA. RelAhepΔ mice and control littermates received intratracheal instillations of S. pneumoniae 24 h before the livers were harvested for RNA extraction. Quantitative RT-PCR analysis

    Techniques Used: Mutagenesis, Mouse Assay, RNA Extraction, Quantitative RT-PCR

    22) Product Images from "Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection"

    Article Title: Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection

    Journal: Experimental gerontology

    doi: 10.1016/j.exger.2017.11.010

    Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P
    Figure Legend Snippet: Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P

    Techniques Used: Infection, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P
    Figure Legend Snippet: Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P

    Techniques Used: Mouse Assay, Infection

    23) Product Images from "The Metabolic Sensor GPR43 Receptor Plays a Role in the Control of Klebsiella pneumoniae Infection in the Lung"

    Article Title: The Metabolic Sensor GPR43 Receptor Plays a Role in the Control of Klebsiella pneumoniae Infection in the Lung

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2018.00142

    The onset of inflammatory cells in the lung of Gpr43 −/− mice 48 h after Klebsiella pneumoniae infection. (A) Number of total cells infiltration, (B) neutrophils, and (C) mononuclear cells in airway spaces recovered using bronchoalveolar lavage (BAL) were counted at different time points (0–48 h) after intratracheal inoculation of 1 × 10 6 CFU of Kp . (D) Myeloperoxidase activity in homogenized lungs harvested from wild-type and Gpr43 −/− mice. (E) IL-1β levels and (F) TNF-α levels were measured by ELISA in BAL supernatant harvested from wild-type and Gpr43 −/− mice 48 h after Kp infection. Results are presented as mean ± SEM ( n = 6). *Statistical difference ( P
    Figure Legend Snippet: The onset of inflammatory cells in the lung of Gpr43 −/− mice 48 h after Klebsiella pneumoniae infection. (A) Number of total cells infiltration, (B) neutrophils, and (C) mononuclear cells in airway spaces recovered using bronchoalveolar lavage (BAL) were counted at different time points (0–48 h) after intratracheal inoculation of 1 × 10 6 CFU of Kp . (D) Myeloperoxidase activity in homogenized lungs harvested from wild-type and Gpr43 −/− mice. (E) IL-1β levels and (F) TNF-α levels were measured by ELISA in BAL supernatant harvested from wild-type and Gpr43 −/− mice 48 h after Kp infection. Results are presented as mean ± SEM ( n = 6). *Statistical difference ( P

    Techniques Used: Mouse Assay, Infection, Activity Assay, Enzyme-linked Immunosorbent Assay

    Oral Treatment with the Gpr43 metabolic ligand, acetate in the lung 48 h after Klebsiella pneumoniae ( Kp ) infection. (A) Survival rate of mice pretreated for 5 days with acetate were observed during 7 days following intratracheal Kp (1 × 10 6 CFU). (B) Bacterial burden in the airways was assessed in lung 48 h post- Kp infection. (C) Myeloperoxidase activity in homogenized lungs harvested from wild-type and wild-type pretreated for 5 days with acetate. (D) Number of total infiltrated cells, (E) neutrophils, and (F) mononuclear cells in airway spaces recovered using bronchoalveolar lavage were counted at 48 h post- Kp infection. (G) Representative photographs of H E-stained sections of lung from wild-type mice control [phosphate-buffered saline (PBS)], wild-type mice Kp infected, and wild-type pretreated 5 days with acetate Kp infected (acetate). Scale bar—400 mm. (H) Number of neutrophils from posttreatment mice with acetate (150 mM) by gavage 24 h after Kp infection. Results are expressed as mean ± SEM of 8–10 mice per group. *Statistical difference ( P
    Figure Legend Snippet: Oral Treatment with the Gpr43 metabolic ligand, acetate in the lung 48 h after Klebsiella pneumoniae ( Kp ) infection. (A) Survival rate of mice pretreated for 5 days with acetate were observed during 7 days following intratracheal Kp (1 × 10 6 CFU). (B) Bacterial burden in the airways was assessed in lung 48 h post- Kp infection. (C) Myeloperoxidase activity in homogenized lungs harvested from wild-type and wild-type pretreated for 5 days with acetate. (D) Number of total infiltrated cells, (E) neutrophils, and (F) mononuclear cells in airway spaces recovered using bronchoalveolar lavage were counted at 48 h post- Kp infection. (G) Representative photographs of H E-stained sections of lung from wild-type mice control [phosphate-buffered saline (PBS)], wild-type mice Kp infected, and wild-type pretreated 5 days with acetate Kp infected (acetate). Scale bar—400 mm. (H) Number of neutrophils from posttreatment mice with acetate (150 mM) by gavage 24 h after Kp infection. Results are expressed as mean ± SEM of 8–10 mice per group. *Statistical difference ( P

    Techniques Used: Infection, Mouse Assay, Activity Assay, Staining

    In vitro neutrophils analyses of phagocytosis and cytokines production after Klebsiella pneumoniae ( Kp ) exposure. (A) Bone marrow neutrophils from wild type and Gpr43 −/− were incubated with opsonized-pHRodo-marked Kp (MOI 1:1) and acetate (1 mM) for 120 min. Samples were analyzed by flow cytometer, and the mean fluorescence of bacteria within the cells were analyzed. (B) IL-10 and (C) IL-1β were measured in supernatant from neutrophils by ELISA. Results are presented as mean ± SEM ( n =6). C− = without bacteria C+ = with bacteria. *Statistical difference ( P
    Figure Legend Snippet: In vitro neutrophils analyses of phagocytosis and cytokines production after Klebsiella pneumoniae ( Kp ) exposure. (A) Bone marrow neutrophils from wild type and Gpr43 −/− were incubated with opsonized-pHRodo-marked Kp (MOI 1:1) and acetate (1 mM) for 120 min. Samples were analyzed by flow cytometer, and the mean fluorescence of bacteria within the cells were analyzed. (B) IL-10 and (C) IL-1β were measured in supernatant from neutrophils by ELISA. Results are presented as mean ± SEM ( n =6). C− = without bacteria C+ = with bacteria. *Statistical difference ( P

    Techniques Used: In Vitro, Incubation, Flow Cytometry, Cytometry, Fluorescence, Enzyme-linked Immunosorbent Assay

    Gpr43 −/− mice are more susceptible to Gram-negative [ Klebsiella pneumoniae ( Kp )] lung infection. (A) Survival rate of mice were observed during 7 days following intratracheal Kp (1 × 10 6 CFU). (B) Weight loss infection post- Kp infection. (C) Bacterial burden in the airways was assessed in bronchoalveolar fluid [bronchoalveolar lavage (BAL)] 48 h post- Kp infection. (D) Representative photographs of H E-stained sections of lung from wild-type mice control [WT phosphate-buffered saline (PBS)], wild-type mice Kp infected (WT INF), Gpr43 −/− mice control (Gpr43 −/− PBS) and Gpr43 −/− mice infected (Gpr43 −/− INF). Scale bar—400 mm. Results are expressed as mean ± SEM of 8–10 mice per group. *Statistical difference ( P
    Figure Legend Snippet: Gpr43 −/− mice are more susceptible to Gram-negative [ Klebsiella pneumoniae ( Kp )] lung infection. (A) Survival rate of mice were observed during 7 days following intratracheal Kp (1 × 10 6 CFU). (B) Weight loss infection post- Kp infection. (C) Bacterial burden in the airways was assessed in bronchoalveolar fluid [bronchoalveolar lavage (BAL)] 48 h post- Kp infection. (D) Representative photographs of H E-stained sections of lung from wild-type mice control [WT phosphate-buffered saline (PBS)], wild-type mice Kp infected (WT INF), Gpr43 −/− mice control (Gpr43 −/− PBS) and Gpr43 −/− mice infected (Gpr43 −/− INF). Scale bar—400 mm. Results are expressed as mean ± SEM of 8–10 mice per group. *Statistical difference ( P

    Techniques Used: Mouse Assay, Infection, Staining

    24) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    25) Product Images from "Evidence for the Presence of Simkania negevensis in Drinking Water and in Reclaimed Wastewater in Israel"

    Article Title: Evidence for the Presence of Simkania negevensis in Drinking Water and in Reclaimed Wastewater in Israel

    Journal: Applied and Environmental Microbiology

    doi: 10.1128/AEM.70.6.3346-3351.2004

    Specificity of MEIA with respect to members of the Chlamydiaceae . Duplicate samples of 100-μl volumes of twofold dilutions of bacterial suspensions were applied to each of the membranes, as indicated. Sn, S. negevensis ; Ct, C. trachomatis L2; Cpn, C. pneumoniae . Starting concentrations were as follows: S. negevensis , 4 × 10 4 organisms per dot; C. trachomatis L2, 2.5 × 10 4 organisms per dot; C. pneumoniae , 2.5 × 10 4 organisms per dot. Upper left panel, rabbit anti- S. negevensis serum used at 1:20,000; upper right panel, rabbit anti- C. trachomatis serum used at 1:20,000; lower left panel, rabbit anti- C. pneumoniae serum used at 1:20,000; lower right panel, serum diluent with secondary antibody only.
    Figure Legend Snippet: Specificity of MEIA with respect to members of the Chlamydiaceae . Duplicate samples of 100-μl volumes of twofold dilutions of bacterial suspensions were applied to each of the membranes, as indicated. Sn, S. negevensis ; Ct, C. trachomatis L2; Cpn, C. pneumoniae . Starting concentrations were as follows: S. negevensis , 4 × 10 4 organisms per dot; C. trachomatis L2, 2.5 × 10 4 organisms per dot; C. pneumoniae , 2.5 × 10 4 organisms per dot. Upper left panel, rabbit anti- S. negevensis serum used at 1:20,000; upper right panel, rabbit anti- C. trachomatis serum used at 1:20,000; lower left panel, rabbit anti- C. pneumoniae serum used at 1:20,000; lower right panel, serum diluent with secondary antibody only.

    Techniques Used:

    26) Product Images from "Lipoprotein Lipase and Hydrofluoric Acid Deactivate Both Bacterial Lipoproteins and Lipoteichoic Acids, but Platelet-Activating Factor-Acetylhydrolase Degrades Only Lipoteichoic Acids ▿"

    Article Title: Lipoprotein Lipase and Hydrofluoric Acid Deactivate Both Bacterial Lipoproteins and Lipoteichoic Acids, but Platelet-Activating Factor-Acetylhydrolase Degrades Only Lipoteichoic Acids ▿

    Journal: Clinical and Vaccine Immunology : CVI

    doi: 10.1128/CVI.00115-09

    TNF-α production by RAW 264.7 cells in response to the butanol extracts (A and B) or the culture supernatants (C and D) from S. pneumoniae (A and C) and S. aureus (B and D). The amounts of the extracts or culture supernatants used for stimulation are indicated as percentages along the x axis. The stimulants were treated with nothing (black bars) or with 30 μg/ml of PAF-AH (white bars). White bars with 0% stimulants represent data for cells stimulated with PAF-AH alone. Bars indicate the means of results for triplicate wells in a representative experiment. Error bars indicate standard deviations. Statistical significance was calculated using one-way ANOVA: **, P
    Figure Legend Snippet: TNF-α production by RAW 264.7 cells in response to the butanol extracts (A and B) or the culture supernatants (C and D) from S. pneumoniae (A and C) and S. aureus (B and D). The amounts of the extracts or culture supernatants used for stimulation are indicated as percentages along the x axis. The stimulants were treated with nothing (black bars) or with 30 μg/ml of PAF-AH (white bars). White bars with 0% stimulants represent data for cells stimulated with PAF-AH alone. Bars indicate the means of results for triplicate wells in a representative experiment. Error bars indicate standard deviations. Statistical significance was calculated using one-way ANOVA: **, P

    Techniques Used:

    27) Product Images from "Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection"

    Article Title: Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection

    Journal: Experimental gerontology

    doi: 10.1016/j.exger.2017.11.010

    Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P
    Figure Legend Snippet: Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P

    Techniques Used: Infection, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P
    Figure Legend Snippet: Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P

    Techniques Used: Mouse Assay, Infection

    28) Product Images from "Nested PCR-Linked Capillary Electrophoresis and Single-Strand Conformation Polymorphisms for Detection of Macrolide-Resistant Mycoplasma pneumoniae in Beijing, China ▿"

    Article Title: Nested PCR-Linked Capillary Electrophoresis and Single-Strand Conformation Polymorphisms for Detection of Macrolide-Resistant Mycoplasma pneumoniae in Beijing, China ▿

    Journal: Journal of Clinical Microbiology

    doi: 10.1128/JCM.00400-10

    Superimposed CE electropherograms of ssDNA 303 (A) and ssDNA 342 (B) from mutated and wild-type samples. Shown are electropherograms of ssDNA 303 from a mutated clinical specimen (line a), the M. pneumoniae reference strain (line b), and a wild-type isolate (line c) and of ssDNA 342 from a mutated clinical specimen (line 1), the M. pneumoniae reference strain (line 2), and a wild-type isolate (line 3).
    Figure Legend Snippet: Superimposed CE electropherograms of ssDNA 303 (A) and ssDNA 342 (B) from mutated and wild-type samples. Shown are electropherograms of ssDNA 303 from a mutated clinical specimen (line a), the M. pneumoniae reference strain (line b), and a wild-type isolate (line c) and of ssDNA 342 from a mutated clinical specimen (line 1), the M. pneumoniae reference strain (line 2), and a wild-type isolate (line 3).

    Techniques Used:

    29) Product Images from "Quantitative analysis of pathogens in the lower respiratory tract of patients with chronic obstructive pulmonary disease"

    Article Title: Quantitative analysis of pathogens in the lower respiratory tract of patients with chronic obstructive pulmonary disease

    Journal: BMC Pulmonary Medicine

    doi: 10.1186/s12890-015-0094-z

    Bacterial load in PSB and BALF samples obtained from COPD patients and healthy subjects. The burden of Staphylococcus aureus , Klebsiella pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis in PSB samples ( a ), and of Klebsiella pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis in BALF samples ( b ) obtained from AECOPD patients compared with stable COPD patients. All data are expressed as means ± SEM. * P
    Figure Legend Snippet: Bacterial load in PSB and BALF samples obtained from COPD patients and healthy subjects. The burden of Staphylococcus aureus , Klebsiella pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis in PSB samples ( a ), and of Klebsiella pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis in BALF samples ( b ) obtained from AECOPD patients compared with stable COPD patients. All data are expressed as means ± SEM. * P

    Techniques Used:

    Standard curves for Staphylococcus aureus , Klebsiella pneumoniae , Streptococcus pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis assays. Mean threshold cycle (CT) value ± SEM of three replicates (three per run) for each reaction is shown in relation to the log of CFU calculated per 20 μl of reaction mixture. Relative correlation coefficient ( R 2 ) and linear regression equations are reported in the legend
    Figure Legend Snippet: Standard curves for Staphylococcus aureus , Klebsiella pneumoniae , Streptococcus pneumoniae , Pseudomonos aeruginosa , Haemophilus influenzeae , and Moraxella catarrhalis assays. Mean threshold cycle (CT) value ± SEM of three replicates (three per run) for each reaction is shown in relation to the log of CFU calculated per 20 μl of reaction mixture. Relative correlation coefficient ( R 2 ) and linear regression equations are reported in the legend

    Techniques Used:

    30) Product Images from "Alveolar macrophage development in mice requires L-plastin for cellular localization in alveoli"

    Article Title: Alveolar macrophage development in mice requires L-plastin for cellular localization in alveoli

    Journal: Blood

    doi: 10.1182/blood-2016-03-705962

    Impaired pneumococcal clearance and increased pneumococcal dissemination in CD11c.Cre pos -LPL fl/fl and CD11c.Cre pos -LPL fl/+ mice. (A) Mice of indicated genotypes were challenged via intratracheal injection of 5 × 10 4 colony-forming units (cfu) of S pneumoniae serotype 3. After 24 hours, bacterial colony-forming units in harvested BAL fluid were determined by serial dilution. Number of mice shown along the x-axis; data combined from at least 3 independent experiments. (B) Percentage of mice with positive blood cultures following pulmonary pneumococcal infection.
    Figure Legend Snippet: Impaired pneumococcal clearance and increased pneumococcal dissemination in CD11c.Cre pos -LPL fl/fl and CD11c.Cre pos -LPL fl/+ mice. (A) Mice of indicated genotypes were challenged via intratracheal injection of 5 × 10 4 colony-forming units (cfu) of S pneumoniae serotype 3. After 24 hours, bacterial colony-forming units in harvested BAL fluid were determined by serial dilution. Number of mice shown along the x-axis; data combined from at least 3 independent experiments. (B) Percentage of mice with positive blood cultures following pulmonary pneumococcal infection.

    Techniques Used: Mouse Assay, Injection, Serial Dilution, Infection

    31) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    32) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    33) Product Images from "Effects of inhaled CO administration on acute lung injury in baboons with pneumococcal pneumonia"

    Article Title: Effects of inhaled CO administration on acute lung injury in baboons with pneumococcal pneumonia

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00240.2015

    Delivery of inhaled CO at 200 ppm for 1 h and carboxyhemoglobin (COHb). A and B : adult male baboons ( n = 5) were inoculated with S. pneumoniae (10 8 -10 9 CFU) and given inhaled CO gas (200 ppm) through the ventilator circuit at 24 or 48 h postinoculation
    Figure Legend Snippet: Delivery of inhaled CO at 200 ppm for 1 h and carboxyhemoglobin (COHb). A and B : adult male baboons ( n = 5) were inoculated with S. pneumoniae (10 8 -10 9 CFU) and given inhaled CO gas (200 ppm) through the ventilator circuit at 24 or 48 h postinoculation

    Techniques Used:

    34) Product Images from "Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection"

    Article Title: Decreased NLRP3 Inflammasome Expression in Aged Lung may contribute to Increased Susceptibility to Secondary Streptococcus pneumoniae Infection

    Journal: Experimental gerontology

    doi: 10.1016/j.exger.2017.11.010

    Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P
    Figure Legend Snippet: Dysregulated TLR Signaling in Aged Lung in Response to Secondary S. pneumoniae Infection (A–C) Young (2 months) and aged (19 months) male and female BALB/c mice received influenza (day 0: PR8, 10 3 PFU-INF) prior to secondary instillation with S. pneumoniae (day 14, 10 3 CFU, ATCC 6303- S. pne ). Lung tissue was collected at 24 hours post-secondary S. pneumoniae infection. Young and aged lung from influenza-infected mice (day 14 post infection) at 24 hours post PBS instillation were used as calibrator samples. mRNA expression values in young and aged lung in response to secondary S. pneumoniae infection are relative to age-matched calibrator samples. (A) Changes in lung mRNA expression of TLR1 (P=0.0011), TLR2, TLR4, TLR6 (P=0.0223), and TLR9 (P=0.0041) in young and aged lung was assessed by real time PCR. (B) Changes in mRNA expression of NOD1 (P=0.0201), NOD2 (P

    Techniques Used: Infection, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P
    Figure Legend Snippet: Increased Morbidity and Mortality of Aged Mice in Response to Secondary S. pneumoniae Infection (A) Experimental layout of primary influenza (day 0 instillation with 10 3 PFU influenza A/PR/8/34 H1N1 (PR8) followed by day 14 instillation with PBS), primary S. pneumoniae (day 0 instillation with PBS followed by day 14 instillation with 10 3 CFU of ATCC 6303), and secondary S. pneumoniae (day 0 instillation with PR8 10 3 PFU followed by day 14 instillation with 10 3 CFU of ATCC 6303) infection in young (2 months) and aged (19 months) adult male and female BALB/c mice. (B) Clinical scores (0hr: P=0.0005, 24hr: P=0.0013, and 48hr: P=0.0073), (C) weight changed (P

    Techniques Used: Mouse Assay, Infection

    35) Product Images from "Pharmacodynamic activity of the lantibiotic MU1140"

    Article Title: Pharmacodynamic activity of the lantibiotic MU1140

    Journal: International journal of antimicrobial agents

    doi: 10.1016/j.ijantimicag.2008.07.028

    Bactericidal activity of MU1140 against Streptococcus pneumoniae strain ATCC 49619.◆, control; ■, 0.5× MIC; ▲, 1× MIC; |, 2× MIC; ▽, 4× MIC; — , 8× MIC. MIC, minimum inhibitory
    Figure Legend Snippet: Bactericidal activity of MU1140 against Streptococcus pneumoniae strain ATCC 49619.◆, control; ■, 0.5× MIC; ▲, 1× MIC; |, 2× MIC; ▽, 4× MIC; — , 8× MIC. MIC, minimum inhibitory

    Techniques Used: Activity Assay

    MU1140 minimum inhibitory concentrations (MICs) after 21 subculturing events for multidrug-resistant Staphylococcus aureus (◆) and Streptococcus pneumoniae (▽). Decrease in susceptibility after repeated subculturing in subinhibitory MU1140
    Figure Legend Snippet: MU1140 minimum inhibitory concentrations (MICs) after 21 subculturing events for multidrug-resistant Staphylococcus aureus (◆) and Streptococcus pneumoniae (▽). Decrease in susceptibility after repeated subculturing in subinhibitory MU1140

    Techniques Used: Subculturing Assay

    36) Product Images from "NLRP3 inflammasome activation in aged macrophages is diminished during Streptococcus pneumoniae infection"

    Article Title: NLRP3 inflammasome activation in aged macrophages is diminished during Streptococcus pneumoniae infection

    Journal: American Journal of Physiology - Lung Cellular and Molecular Physiology

    doi: 10.1152/ajplung.00393.2017

    Similar NF-κB expression in young and aged macrophages in response to S. pneumoniae . A – D : young and aged macrophages were stimulated with TLR-1/2 (Pam3CSK4, 100 ng/ml), TLR-2 (HKLM, 10 7 cells/ml), TLR-3 (HMW poly I:C, 10 ng/ml), TLR-3 (LMW poly I:C, 30 ng/ml), TLR-4 (LPS-EK, 10 ng/ml), TLR-5 (FLA-st, 100 ng/ml), TLR-6 (FSL-1 10 ng/ml), TLR-7 (ssRNA40, 1 μg/ml), or TLR-9 (ODN1826, 1 μM) agonists. TNF-α (TLR-1/2: t -test, P = 0.0446; TLR-3 (HMW): t -test, * P = 0.0114; TLR-3 (LMW): t -test, *** P = 0.0006; TLR-9: t -test, * P = 0.0462) ( A ) and IL-1β (TLR-1/2: t -test, ** P = 0.0037; TLR-2: t -test, ** P = 0.0016; TLR-4: t -test, * P = 0.0155; TLR-6: t -test, ** P = 0.0097; TLR-9: t -test, *** P = 0.0002) ( B ) production was quantified by ELISA. C and D : cytosolic and nuclear protein extracts were isolated from young and aged macrophages at 2 h post- S. pneumoniae (50 CFU) infection. NF-κB ( t -test, **** P
    Figure Legend Snippet: Similar NF-κB expression in young and aged macrophages in response to S. pneumoniae . A – D : young and aged macrophages were stimulated with TLR-1/2 (Pam3CSK4, 100 ng/ml), TLR-2 (HKLM, 10 7 cells/ml), TLR-3 (HMW poly I:C, 10 ng/ml), TLR-3 (LMW poly I:C, 30 ng/ml), TLR-4 (LPS-EK, 10 ng/ml), TLR-5 (FLA-st, 100 ng/ml), TLR-6 (FSL-1 10 ng/ml), TLR-7 (ssRNA40, 1 μg/ml), or TLR-9 (ODN1826, 1 μM) agonists. TNF-α (TLR-1/2: t -test, P = 0.0446; TLR-3 (HMW): t -test, * P = 0.0114; TLR-3 (LMW): t -test, *** P = 0.0006; TLR-9: t -test, * P = 0.0462) ( A ) and IL-1β (TLR-1/2: t -test, ** P = 0.0037; TLR-2: t -test, ** P = 0.0016; TLR-4: t -test, * P = 0.0155; TLR-6: t -test, ** P = 0.0097; TLR-9: t -test, *** P = 0.0002) ( B ) production was quantified by ELISA. C and D : cytosolic and nuclear protein extracts were isolated from young and aged macrophages at 2 h post- S. pneumoniae (50 CFU) infection. NF-κB ( t -test, **** P

    Techniques Used: Expressing, Enzyme-linked Immunosorbent Assay, Isolation, Infection

    Endoplasmic reticulum (ER) stress in the aged lung is enhanced during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in BIP/GRP78 expression in lung homogenates was calculated at baseline and at 24, 48, and 72 h postinfection (24 h: t -test, * P = 0.0189; 48 h: t -test, ** P = 0.0028; 72 h, t -test: ** P = 0.0020). B and C : inositol-requiring enzyme 1 (IRE1) and activating transcription factor 6 (ATF6) ( B ) and PKR-like ER resident kinase (PERK) ( C ) mRNA expression was quantified by real-time PCR at 24 h postinfection (IRE1: t -test, * P = 0.0119; PERK: t -test, ** P
    Figure Legend Snippet: Endoplasmic reticulum (ER) stress in the aged lung is enhanced during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in BIP/GRP78 expression in lung homogenates was calculated at baseline and at 24, 48, and 72 h postinfection (24 h: t -test, * P = 0.0189; 48 h: t -test, ** P = 0.0028; 72 h, t -test: ** P = 0.0020). B and C : inositol-requiring enzyme 1 (IRE1) and activating transcription factor 6 (ATF6) ( B ) and PKR-like ER resident kinase (PERK) ( C ) mRNA expression was quantified by real-time PCR at 24 h postinfection (IRE1: t -test, * P = 0.0119; PERK: t -test, ** P

    Techniques Used: Infection, Mouse Assay, Expressing, Real-time Polymerase Chain Reaction

    AIM2 inflammasome responses are not essential for caspase-1 activation or IL-1β production in response to a serotype 3 strain of S. pneumoniae . Young wild-type (WT) and Aim2 −/− macrophages were cultured with S. pneumoniae ( S. pne ) (50 CFU) and mRNA and protein expression was examined at 4 and 24 h postinfection. A : changes in TLR- 1, 2, 4, 6, and 9, and MYD88 were quantified by real-time PCR. B : mRNA expression of NOD1, NOD2 (4 h: t -test, P
    Figure Legend Snippet: AIM2 inflammasome responses are not essential for caspase-1 activation or IL-1β production in response to a serotype 3 strain of S. pneumoniae . Young wild-type (WT) and Aim2 −/− macrophages were cultured with S. pneumoniae ( S. pne ) (50 CFU) and mRNA and protein expression was examined at 4 and 24 h postinfection. A : changes in TLR- 1, 2, 4, 6, and 9, and MYD88 were quantified by real-time PCR. B : mRNA expression of NOD1, NOD2 (4 h: t -test, P

    Techniques Used: Activation Assay, Cell Culture, Expressing, Real-time Polymerase Chain Reaction

    Treatment with TUDCA rescues NLRP3 inflammasome activation in the aged lung during S. pneumoniae infection. A – E ; young (2 mo) and aged (18 mo) male and female BALB/c mice received saline or TUDCA (500 mg·kg −1 ·day −1 ) via intraperitoneal injection before intranasal instillation with saline or 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : gene expression in lung tissue collected from young, aged, and TUDCA-treated aged mice at 24 h postinfection was quantified by real-time PCR using inflammasome specific primer assays (SABiosciences). ASC, NLRP3, and pro-caspase-1 mRNA expression was evaluated by real-time PCR (ASC: t -test, **** P
    Figure Legend Snippet: Treatment with TUDCA rescues NLRP3 inflammasome activation in the aged lung during S. pneumoniae infection. A – E ; young (2 mo) and aged (18 mo) male and female BALB/c mice received saline or TUDCA (500 mg·kg −1 ·day −1 ) via intraperitoneal injection before intranasal instillation with saline or 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : gene expression in lung tissue collected from young, aged, and TUDCA-treated aged mice at 24 h postinfection was quantified by real-time PCR using inflammasome specific primer assays (SABiosciences). ASC, NLRP3, and pro-caspase-1 mRNA expression was evaluated by real-time PCR (ASC: t -test, **** P

    Techniques Used: Activation Assay, Infection, Mouse Assay, Injection, Expressing, Real-time Polymerase Chain Reaction

    Macrophage and lung Toll-like receptor (TLR) expression in response to S. pneumoniae . A – C : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in mRNA expression of TLR-1 ( t -test, * P = 0.01354), 2 ( t -test, ** P = 0.0082), 4 ( t -test, * P = 0.0487), 6, and 9 ( t -test, * P = 0.0144) in young and aged lungs was evaluated by real-time PCR. B : changes in mRNA expression of NOD1 ( t -test, * P = 0.0054), NOD2 ( t -test, ** P = 0.015), RIP2, and NF-κB at 24 h postinfection were quantified by real-time PCR. C : inflammatory cytokine expression was investigated in young and aged lungs, and changes in pro-IL-1β, pro-IL-18, IL-6 ( t -test, **** P
    Figure Legend Snippet: Macrophage and lung Toll-like receptor (TLR) expression in response to S. pneumoniae . A – C : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 CFU of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : changes in mRNA expression of TLR-1 ( t -test, * P = 0.01354), 2 ( t -test, ** P = 0.0082), 4 ( t -test, * P = 0.0487), 6, and 9 ( t -test, * P = 0.0144) in young and aged lungs was evaluated by real-time PCR. B : changes in mRNA expression of NOD1 ( t -test, * P = 0.0054), NOD2 ( t -test, ** P = 0.015), RIP2, and NF-κB at 24 h postinfection were quantified by real-time PCR. C : inflammatory cytokine expression was investigated in young and aged lungs, and changes in pro-IL-1β, pro-IL-18, IL-6 ( t -test, **** P

    Techniques Used: Expressing, Mouse Assay, Real-time Polymerase Chain Reaction

    Treatment with tauroursodeoxycholic acid improves NLRP3 inflammasome activation in aged macrophages during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male or female BALB/c macrophages were pretreated with tauroursodeoxycholic acid (TUDCA) (100 μM) for 24 h before culture with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : mRNA expression of ATF6, IRE1, and PERK in untreated and TUDCA-treated aged macrophages at 24 h postinfection was quantified by real-time PCR (IRE1: t -test, *** P = 0.0008). B : IRE1 phosphorylation in untreated and TUDCA pretreated aged macrophages at 24 h postinfection was examined by Western blot. C : XBP1 splicing was evaluated by PCR and % of XBP1 splicing was quantified ( t -test, ** P = 0.0032). D : cytotoxicity in young and aged macrophages in response to TUDCA was examined at 24 h post- S. pneumoniae infection ( t -test, ** P
    Figure Legend Snippet: Treatment with tauroursodeoxycholic acid improves NLRP3 inflammasome activation in aged macrophages during S. pneumoniae infection. Young (2 mo) and aged (18 mo) male or female BALB/c macrophages were pretreated with tauroursodeoxycholic acid (TUDCA) (100 μM) for 24 h before culture with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ) (ATCC 6303). A : mRNA expression of ATF6, IRE1, and PERK in untreated and TUDCA-treated aged macrophages at 24 h postinfection was quantified by real-time PCR (IRE1: t -test, *** P = 0.0008). B : IRE1 phosphorylation in untreated and TUDCA pretreated aged macrophages at 24 h postinfection was examined by Western blot. C : XBP1 splicing was evaluated by PCR and % of XBP1 splicing was quantified ( t -test, ** P = 0.0032). D : cytotoxicity in young and aged macrophages in response to TUDCA was examined at 24 h post- S. pneumoniae infection ( t -test, ** P

    Techniques Used: Activation Assay, Infection, Expressing, Real-time Polymerase Chain Reaction, Western Blot, Polymerase Chain Reaction

    NLRP3 expression in the aged lung is necessary for survival during Streptococcus pneumoniae . A , D – H : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 colony-forming untis (CFU) of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : hematoxylin and eosin (H E) staining of lung tissue from young and aged control and S. pneumoniae -infected mice collected at 72 h postinfection. B and C : young (2 mo) and aged (17 mo) male and female wild-type C57BL/6 or C57Bl/6-NLRP3 −/− mice were instilled with 1 × 10 3 CFU of S. pneumoniae . Survival ( B ) (Mantel-Cox: P = 0.0026) and bacterial titer ( C ) in lung was examined in lung homogenates at 24 h postinfection ( t -test, ** P
    Figure Legend Snippet: NLRP3 expression in the aged lung is necessary for survival during Streptococcus pneumoniae . A , D – H : young (2 mo) and aged (18 mo) male and female BALB/c mice received 1 × 10 3 colony-forming untis (CFU) of S. pneumoniae ( S. pne ) (ATCC 6303) or saline via intranasal instillation. A : hematoxylin and eosin (H E) staining of lung tissue from young and aged control and S. pneumoniae -infected mice collected at 72 h postinfection. B and C : young (2 mo) and aged (17 mo) male and female wild-type C57BL/6 or C57Bl/6-NLRP3 −/− mice were instilled with 1 × 10 3 CFU of S. pneumoniae . Survival ( B ) (Mantel-Cox: P = 0.0026) and bacterial titer ( C ) in lung was examined in lung homogenates at 24 h postinfection ( t -test, ** P

    Techniques Used: Expressing, Mouse Assay, Staining, Infection

    Diminished NLRP3 inflammasome activation in aged macrophages in response to S. pneumoniae . Young (2 mo) and aged (18 mo) male and female BALB/c macrophages were treated with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ). A : baseline and S. pneumoniae upregulated pro-caspase-1, NLRP3, and ASC mRNA expression was evaluated by real-time PCR ( t -test, *** P
    Figure Legend Snippet: Diminished NLRP3 inflammasome activation in aged macrophages in response to S. pneumoniae . Young (2 mo) and aged (18 mo) male and female BALB/c macrophages were treated with media alone or media containing 50 CFU of S. pneumoniae ( S. pne ). A : baseline and S. pneumoniae upregulated pro-caspase-1, NLRP3, and ASC mRNA expression was evaluated by real-time PCR ( t -test, *** P

    Techniques Used: Activation Assay, Expressing, Real-time Polymerase Chain Reaction

    37) Product Images from "Signature patterns revealed by microarray analyses of mice infected with influenza virus A and Streptococcus pneumoniae"

    Article Title: Signature patterns revealed by microarray analyses of mice infected with influenza virus A and Streptococcus pneumoniae

    Journal: Microbes and Infection

    doi: 10.1016/j.micinf.2006.04.018

    Signature patterns of gene expression in mice in response to influenza virus A/PR/8/34 and Streptococcus pneumonia. Twenty-eight differentially expressed genes were selected to show the difference of gene expression patterns between mice infected with influenza virus A/PR/8/34 and Streptococcus pneumoniae at day 1. The relative fold changes for each gene were used for comparison.
    Figure Legend Snippet: Signature patterns of gene expression in mice in response to influenza virus A/PR/8/34 and Streptococcus pneumonia. Twenty-eight differentially expressed genes were selected to show the difference of gene expression patterns between mice infected with influenza virus A/PR/8/34 and Streptococcus pneumoniae at day 1. The relative fold changes for each gene were used for comparison.

    Techniques Used: Expressing, Mouse Assay, Infection

    Distribution of functions of genes that were differentially expressed in response to infection of mice with influenza virus A/PR/8/34 and Streptococcus pneumoniae.
    Figure Legend Snippet: Distribution of functions of genes that were differentially expressed in response to infection of mice with influenza virus A/PR/8/34 and Streptococcus pneumoniae.

    Techniques Used: Infection, Mouse Assay

    38) Product Images from "Dysregulation of Macrophage Activation Profiles by Engineered Nanoparticles"

    Article Title: Dysregulation of Macrophage Activation Profiles by Engineered Nanoparticles

    Journal: ACS nano

    doi: 10.1021/nn402145t

    Effects of ENP Pretreatment on Macrophage Phagocytosis of S. pneumoniae
    Figure Legend Snippet: Effects of ENP Pretreatment on Macrophage Phagocytosis of S. pneumoniae

    Techniques Used:

    39) Product Images from "L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection"

    Article Title: L-Plastin Is Essential for Alveolar Macrophage Production and Control of Pulmonary Pneumococcal Infection

    Journal: Infection and Immunity

    doi: 10.1128/IAI.01199-13

    Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.
    Figure Legend Snippet: Decreased numbers of alveolar macrophages recovered from BAL fluid of LPL −/− mice. Phagocytosis of pneumococci by alveolar macrophages was unimpaired in LPL −/− mice. (A) Total numbers of alveolar macrophages isolated from BAL fluid of WT (gray circles) and LPL −/− (filled circles) mice, uninfected or infected for the indicated periods of time. (B) Percentages of CD45 + cells that were CD11c high isolated from the BAL fluid of WT (gray circles), LPL −/− (filled circles), and Rag1 −/− (open circles) mice 6 h after i.t. instillation of pneumococci. For panels A and B, each symbol represents data from one mouse, lines represent median values, P values were determined by a Mann-Whitney test, and data were pooled from at least 2 independent experiments per time interval. (C) Representative flow cytometric analysis of numbers of alveolar macrophages (left) and phagocytosis of FITC-labeled S. pneumoniae (SP-FITC) by gated macrophages (right). Alveolar macrophages were assessed by using the phenotypic markers CD45 + and CD11c + ; neutrophils were excluded based on Ly6G + CD11c − staining. A control panel for a WT mouse challenged with PBS is shown at the top. (D) Percentages of alveolar macrophages isolated from BAL fluid of WT or LPL −/− mice that were positive for FITC-labeled pneumococci 3 h after challenge. Each symbol represents one mouse, lines indicate median values, P values were determined by a Mann-Whitney test, and data were from 2 independent experiments.

    Techniques Used: Mouse Assay, Isolation, Infection, MANN-WHITNEY, Flow Cytometry, Labeling, Staining

    LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).
    Figure Legend Snippet: LPL −/− mice are profoundly susceptible to infection with pneumococci and exhibit a very early pneumococcal clearance defect. (A) Age- and gender-matched WT ( n = 9; 5 female and 4 male) and LPL −/− ( n = 10; 5 female and 5 male) mice were infected i.t. with S. pneumoniae serotype 3 strain ATCC 6303 at time zero. A single, nonsterilizing dose of ceftriaxone was given at 24 h (arrow). Mice were monitored for 2 weeks for morbidity and mortality. Data were pooled from 2 independent experiments. P values were determined by a log-rank test. (B to E) Colony counts from BAL fluid (B to E) or blood (D and E) of the indicated mice 3 h (B), 6 h (C), 24 h (D), and 48 h (E) after i.t. instillation of pneumococci. No antibiotic was given for panels B, C, and D, but a single nonsterilizing dose of ceftriaxone was given at 24 h for panel E, to correlate with the treatment data in panel A. Each point represents a single animal, lines represent median values, and P values were determined by using a two-tailed Mann-Whitney test. Data were from 2 (B to E) or 3 (C) independent experiments. For panels B to D, all mice were male. For panel E, mice of each genotype were male ( n = 5) and female ( n = 5).

    Techniques Used: Mouse Assay, Infection, Two Tailed Test, MANN-WHITNEY

    40) Product Images from "Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone"

    Article Title: Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone

    Journal: Scientific Reports

    doi: 10.1038/s41598-018-37438-1

    Mitochondrial Membrane Potential (ΔΨm) is disrupted in Aged Macrophages during S . pneumoniae Infection. Young and aged macrophages were cultured with S . pneumoniae (MOI = 25). ( A ) Cell viability was quantified at 4 and 24 hours post infection using the MT cell viability assay. Light production by metabolically active cells was proportional to the number of live cells in culture (t-test: *** P
    Figure Legend Snippet: Mitochondrial Membrane Potential (ΔΨm) is disrupted in Aged Macrophages during S . pneumoniae Infection. Young and aged macrophages were cultured with S . pneumoniae (MOI = 25). ( A ) Cell viability was quantified at 4 and 24 hours post infection using the MT cell viability assay. Light production by metabolically active cells was proportional to the number of live cells in culture (t-test: *** P

    Techniques Used: Infection, Cell Culture, Viability Assay, Metabolic Labelling, T-Test

    Pirfenidone Treatment Decreases Oxidative Stress in Aged Macrophages in Response to S . pneumoniae . Aged and/or young macrophages were cultured with media alone or media containing pirfenidone (500 μg/ml) for 24 hours prior to culture with S . pneumoniae (MOI = 25). ( A ) At 2 hours post S . pneumoniae infection, changes in cellular ROS levels were examined using H 2 DCFDA (10 μM) and changes in DCF fluorescence was assessed (t-test: *** P
    Figure Legend Snippet: Pirfenidone Treatment Decreases Oxidative Stress in Aged Macrophages in Response to S . pneumoniae . Aged and/or young macrophages were cultured with media alone or media containing pirfenidone (500 μg/ml) for 24 hours prior to culture with S . pneumoniae (MOI = 25). ( A ) At 2 hours post S . pneumoniae infection, changes in cellular ROS levels were examined using H 2 DCFDA (10 μM) and changes in DCF fluorescence was assessed (t-test: *** P

    Techniques Used: Cell Culture, Infection, Fluorescence, T-Test

    Pirfenidone Treatment Improves Antioxidant Responses in Aged Lung during S . pneumoniae Infection. Aged (19 months) male and female BALB/c mice received pirfenidone (280 mg/kg) intraperitoneally 24 hours prior and at the time of infection (1 × 10 6 CFU of S . pneumoniae or saline via intranasal instillation). Lung tissue from aged untreated and pirfenidone treated mice collected was collected at 24 hours post S. pneumoniae infection. ( A ) Superoxide generation and ΔΨm was examined in aged control and pirfenidone treated lung tissues. Lung cells were collected at 24 hours post saline or S . pneumoniae instillation and evaluated by flow cytometry. Superoxide production was measured using MitoSOX. Young (black line), aged untreated (grey line), and aged pirfenidone treated (blue line). ( B ) Changes in MitoCapture red fluorescence was used to assess ΔΨm. Young (black line), aged untreated (grey line), and aged pirfenidone treated (blue line). ( C , D ) Mean fluorescence intensity (MFI) was assessed for each sample (t-test: *** P
    Figure Legend Snippet: Pirfenidone Treatment Improves Antioxidant Responses in Aged Lung during S . pneumoniae Infection. Aged (19 months) male and female BALB/c mice received pirfenidone (280 mg/kg) intraperitoneally 24 hours prior and at the time of infection (1 × 10 6 CFU of S . pneumoniae or saline via intranasal instillation). Lung tissue from aged untreated and pirfenidone treated mice collected was collected at 24 hours post S. pneumoniae infection. ( A ) Superoxide generation and ΔΨm was examined in aged control and pirfenidone treated lung tissues. Lung cells were collected at 24 hours post saline or S . pneumoniae instillation and evaluated by flow cytometry. Superoxide production was measured using MitoSOX. Young (black line), aged untreated (grey line), and aged pirfenidone treated (blue line). ( B ) Changes in MitoCapture red fluorescence was used to assess ΔΨm. Young (black line), aged untreated (grey line), and aged pirfenidone treated (blue line). ( C , D ) Mean fluorescence intensity (MFI) was assessed for each sample (t-test: *** P

    Techniques Used: Infection, Mouse Assay, Flow Cytometry, Cytometry, Fluorescence, T-Test

    Mitochondrial Membrane Potential (ΔΨm) in Aged Lung Declines during S . pneumoniae Infection. Young (2 months) and aged (19 months) male and female BALB/c mice received 1 × 10 6 CFU of S . pneumoniae (ATCC 6303) or saline via intranasal instillation. Lung tissue from young and aged uninfected and S . pneumoniae infected mice was collected at select time points post infection. ( A ) ΔΨm was examined in lung cells by assessing changes in MitoCapture red and green fluorescence. Dot plots: young (red cells) and aged (blue cells). Histograms: young (black line) and aged (grey line). For histograms, N = 3 are shown for the 24–72 hour time points. ( B ) % of MitoCapture red positive cells and ( C ) mean fluorescence intensity (MFI) was assessed for each sample (t-test: * P
    Figure Legend Snippet: Mitochondrial Membrane Potential (ΔΨm) in Aged Lung Declines during S . pneumoniae Infection. Young (2 months) and aged (19 months) male and female BALB/c mice received 1 × 10 6 CFU of S . pneumoniae (ATCC 6303) or saline via intranasal instillation. Lung tissue from young and aged uninfected and S . pneumoniae infected mice was collected at select time points post infection. ( A ) ΔΨm was examined in lung cells by assessing changes in MitoCapture red and green fluorescence. Dot plots: young (red cells) and aged (blue cells). Histograms: young (black line) and aged (grey line). For histograms, N = 3 are shown for the 24–72 hour time points. ( B ) % of MitoCapture red positive cells and ( C ) mean fluorescence intensity (MFI) was assessed for each sample (t-test: * P

    Techniques Used: Infection, Mouse Assay, Fluorescence, T-Test

    Pirfenidone Treatment Improves Mitochondrial Membrane Potential and Decreases Superoxide Production in Aged Macrophages during S . pneumoniae Infection. Young macrophages were cultured with media alone for 24 hours prior to culture with S . pneumoniae (MOI = 25). Aged macrophages were cultured with media alone or media containing pirfenidone (500 μg/ml) for 24 hours prior to culture with S . pneumoniae (MOI = 25). ( A , B ) ΔΨm was examined in young, aged, and pirfenidone treated aged macrophages by assessing changes in MitoCapture red and green fluorescence in response to 2 hours of S . pneumoniae infection. ( C ) % of MitoCapture red positive cells and ( D ) mean fluorescence intensity (MFI) was assessed for each sample (t-test: * P
    Figure Legend Snippet: Pirfenidone Treatment Improves Mitochondrial Membrane Potential and Decreases Superoxide Production in Aged Macrophages during S . pneumoniae Infection. Young macrophages were cultured with media alone for 24 hours prior to culture with S . pneumoniae (MOI = 25). Aged macrophages were cultured with media alone or media containing pirfenidone (500 μg/ml) for 24 hours prior to culture with S . pneumoniae (MOI = 25). ( A , B ) ΔΨm was examined in young, aged, and pirfenidone treated aged macrophages by assessing changes in MitoCapture red and green fluorescence in response to 2 hours of S . pneumoniae infection. ( C ) % of MitoCapture red positive cells and ( D ) mean fluorescence intensity (MFI) was assessed for each sample (t-test: * P

    Techniques Used: Infection, Cell Culture, Fluorescence, T-Test

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  • 93
    ATCC rvg29 nano bap85 treated groups
    Colony formation unit of S. pneumonia ATCC 49619 in CSF of the mice without any treatment (the negative control group) or treated with <t>Nano-BAs</t> or Penicillin G for various periods of time (A). Quantitative counts of S. pneumonia ATCC 49619 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (B). Survival rate of the PM mice infected with S. pneumonia ATCC49619 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, <t>RVG29-Nano-BAP85,</t> RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (C). Colony formation unit of S. pneumonia 16167 in CSF of the mice without any treatment (the negative control group) or treated with Nano-BAs or Penicillin G for various periods of time (D). Quantitative counts of S. pneumonia 16167 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (E). Survival rate of the PM mice infected with S. pneumonia 16167 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, RVG29-Nano-BAP85, RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (F). Values are mean ± SD of six independent observations. * p
    Rvg29 Nano Bap85 Treated Groups, supplied by ATCC, used in various techniques. Bioz Stars score: 93/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    88
    ATCC hintl 1 binding
    Human IntL-1 binds to S. pneumoniae serotypes producing capsular polysaccharides with terminal vicinal diols. ( a ) Chemical structure of the capsular polysaccharides displayed on the S. pneumoniae serotypes (8, 20, 43, 70) tested. The Gal f residues assumed to mediate <t>hIntL-1</t> cell binding are shown in red and the phosphoglycerol moiety is shown in blue. ( b ) Fluorescence microscopy of hIntL-1 binding to S. pneumoniae serotype 20. Bacteria were treated with Strep-tagged hIntL-1 (15 µg/mL) and an anti-Strep-tag antibody conjugate (red). Cellular DNA was visualized with Hoechst (blue). Left: hIntL-1 marks the surface of serotype 20 bacteria; serotypes 43 and 70 gave similar results; no binding to serotype 8 was detected ( Supplementary Fig. 6a ) Right: EDTA addition abrogates hIntL-1 binding to the bacterial surface, supporting the role for Ca 2+ . Images are representative of > 5 fields of view per sample. Scale bar, 2 µm. ( c,d ) Flow cytometry analysis of Strep-hIntL-1 binding to S. pneumoniae serotypes with an anti-Strep-tag antibody conjugate. In the anti-Strep control sample, recombinant hIntL-1 was omitted. Cells were labeled with propidium iodide. ( c ) Flow cytometry analysis of serotypes 8, 20, 43, and 70; data were collected consecutively with identical instrument settings. ( d ) The dependence of the hIntL-1–carbohydrate interaction on Ca 2+ was tested by adding 10 mM EDTA and ligand selectivity was tested by adding 100 mM glycerol. Data are representative of two independent experiments. For a similar analysis of serotypes 20 and 70 (no binding to serotype 8 was detected), see Supplementary Fig. 6b .
    Hintl 1 Binding, supplied by ATCC, used in various techniques. Bioz Stars score: 88/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    masp 2  (ATCC)
    91
    ATCC masp 2
    Functional role of <t>MASP-2</t> in pneumococcal meningitis mouse model. Kaplan-Meier curve showing increased survival in Masp2 −/− mice during pneumococcal meningitis ( a ). Cytokines and complement levels were measured in brain. Masp2 −/− mice had significantly lower brain levels of IL-1β, IL-10, and TNF-α 30 h after infection ( b – d ). Brain levels of C5b-9 were similar between Masp2 −/− and WT mice ( e ). Data are given as medians and 75th quartile; P values were determined with the Mann-Whitney U test
    Masp 2, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    ATCC pneumococcal polysaccharide serotype 3
    mRNA expression of CD40L in PBMC cultures from 14 patients before and 4 to 6 weeks after immunization with 23-PV compared to mRNA expression in PBMC cultures from four unimmunized controls. Several patient values are very similar, so some lines are superimposed. Cells were stimulated for 5 h with the optimal concentration of pneumococcal polysaccharide serotype 3, 14 or 18C. ELISA-PCR results are expressed as copy numbers.
    Pneumococcal Polysaccharide Serotype 3, supplied by ATCC, used in various techniques. Bioz Stars score: 85/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    Colony formation unit of S. pneumonia ATCC 49619 in CSF of the mice without any treatment (the negative control group) or treated with Nano-BAs or Penicillin G for various periods of time (A). Quantitative counts of S. pneumonia ATCC 49619 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (B). Survival rate of the PM mice infected with S. pneumonia ATCC49619 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, RVG29-Nano-BAP85, RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (C). Colony formation unit of S. pneumonia 16167 in CSF of the mice without any treatment (the negative control group) or treated with Nano-BAs or Penicillin G for various periods of time (D). Quantitative counts of S. pneumonia 16167 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (E). Survival rate of the PM mice infected with S. pneumonia 16167 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, RVG29-Nano-BAP85, RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (F). Values are mean ± SD of six independent observations. * p

    Journal: Drug Delivery

    Article Title: Brain-targeted delivery of PEGylated nano-bacitracin A against Penicillin-sensitive and -resistant Pneumococcal meningitis: formulated with RVG29 and Pluronic® P85 unimers

    doi: 10.1080/10717544.2018.1486473

    Figure Lengend Snippet: Colony formation unit of S. pneumonia ATCC 49619 in CSF of the mice without any treatment (the negative control group) or treated with Nano-BAs or Penicillin G for various periods of time (A). Quantitative counts of S. pneumonia ATCC 49619 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (B). Survival rate of the PM mice infected with S. pneumonia ATCC49619 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, RVG29-Nano-BAP85, RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (C). Colony formation unit of S. pneumonia 16167 in CSF of the mice without any treatment (the negative control group) or treated with Nano-BAs or Penicillin G for various periods of time (D). Quantitative counts of S. pneumonia 16167 per gram of brain tissues of the mice without any treatment (the control group) or treated with Nano-BAs or Penicillin G for 14 days (E). Survival rate of the PM mice infected with S. pneumonia 16167 after treated with Saline, PEGylated Nano-BA12K, Penicillin G, RVG29-Nano-BAP85, RVG29-Nano-BA, Nano-BAP85, and Nano-BA, respectively (F). Values are mean ± SD of six independent observations. * p

    Article Snippet: In contrast, there was no injury found in the kidney of the RVG29 -Nano-BAP85 -treated groups in both S. pneumonia ATCC 49619 and S. pneumonia 16167 infected PM mouse models, indicating low nephrotoxicity and good safety in vivo .

    Techniques: Mouse Assay, Negative Control, Infection

    Human IntL-1 binds to S. pneumoniae serotypes producing capsular polysaccharides with terminal vicinal diols. ( a ) Chemical structure of the capsular polysaccharides displayed on the S. pneumoniae serotypes (8, 20, 43, 70) tested. The Gal f residues assumed to mediate hIntL-1 cell binding are shown in red and the phosphoglycerol moiety is shown in blue. ( b ) Fluorescence microscopy of hIntL-1 binding to S. pneumoniae serotype 20. Bacteria were treated with Strep-tagged hIntL-1 (15 µg/mL) and an anti-Strep-tag antibody conjugate (red). Cellular DNA was visualized with Hoechst (blue). Left: hIntL-1 marks the surface of serotype 20 bacteria; serotypes 43 and 70 gave similar results; no binding to serotype 8 was detected ( Supplementary Fig. 6a ) Right: EDTA addition abrogates hIntL-1 binding to the bacterial surface, supporting the role for Ca 2+ . Images are representative of > 5 fields of view per sample. Scale bar, 2 µm. ( c,d ) Flow cytometry analysis of Strep-hIntL-1 binding to S. pneumoniae serotypes with an anti-Strep-tag antibody conjugate. In the anti-Strep control sample, recombinant hIntL-1 was omitted. Cells were labeled with propidium iodide. ( c ) Flow cytometry analysis of serotypes 8, 20, 43, and 70; data were collected consecutively with identical instrument settings. ( d ) The dependence of the hIntL-1–carbohydrate interaction on Ca 2+ was tested by adding 10 mM EDTA and ligand selectivity was tested by adding 100 mM glycerol. Data are representative of two independent experiments. For a similar analysis of serotypes 20 and 70 (no binding to serotype 8 was detected), see Supplementary Fig. 6b .

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: Human IntL-1 binds to S. pneumoniae serotypes producing capsular polysaccharides with terminal vicinal diols. ( a ) Chemical structure of the capsular polysaccharides displayed on the S. pneumoniae serotypes (8, 20, 43, 70) tested. The Gal f residues assumed to mediate hIntL-1 cell binding are shown in red and the phosphoglycerol moiety is shown in blue. ( b ) Fluorescence microscopy of hIntL-1 binding to S. pneumoniae serotype 20. Bacteria were treated with Strep-tagged hIntL-1 (15 µg/mL) and an anti-Strep-tag antibody conjugate (red). Cellular DNA was visualized with Hoechst (blue). Left: hIntL-1 marks the surface of serotype 20 bacteria; serotypes 43 and 70 gave similar results; no binding to serotype 8 was detected ( Supplementary Fig. 6a ) Right: EDTA addition abrogates hIntL-1 binding to the bacterial surface, supporting the role for Ca 2+ . Images are representative of > 5 fields of view per sample. Scale bar, 2 µm. ( c,d ) Flow cytometry analysis of Strep-hIntL-1 binding to S. pneumoniae serotypes with an anti-Strep-tag antibody conjugate. In the anti-Strep control sample, recombinant hIntL-1 was omitted. Cells were labeled with propidium iodide. ( c ) Flow cytometry analysis of serotypes 8, 20, 43, and 70; data were collected consecutively with identical instrument settings. ( d ) The dependence of the hIntL-1–carbohydrate interaction on Ca 2+ was tested by adding 10 mM EDTA and ligand selectivity was tested by adding 100 mM glycerol. Data are representative of two independent experiments. For a similar analysis of serotypes 20 and 70 (no binding to serotype 8 was detected), see Supplementary Fig. 6b .

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Binding Assay, Fluorescence, Microscopy, Strep-tag, Flow Cytometry, Cytometry, Recombinant, Labeling

    Glycan selectivity of hIntL-1 assessed by glycan microarrays. ( a ) Recombinant hIntL-1 (50 µg/mL) binding to mammalian glycan microarray CFG v5.1 and a furanoside array. The concentrations given for the furanoside array represent those used in the carbohydrate immobilization reaction. Data are presented as the mean ± s.d. ( n =4 technical replicates). The full data set can be found in Supplementary Tables 1 and 2 . ( b ) Recombinant Strep -hIntL-1 (50 µg/mL) binding to microbial glycan array. For glycan array data organized by genus, see Supplementary Fig. 2a . Data are presented as the mean ± s.d. ( n =4 technical replicates). The full data set can be found in Supplementary Table 3 . ( c ) Structural representation of the putative key binding epitopes for hIntL-1 and the non-binding N-acetylneuraminic acid (α-Neu5Ac). A terminal vicinal diol (red) is a common feature of α-Neu5Ac and all of the ligands identified.

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: Glycan selectivity of hIntL-1 assessed by glycan microarrays. ( a ) Recombinant hIntL-1 (50 µg/mL) binding to mammalian glycan microarray CFG v5.1 and a furanoside array. The concentrations given for the furanoside array represent those used in the carbohydrate immobilization reaction. Data are presented as the mean ± s.d. ( n =4 technical replicates). The full data set can be found in Supplementary Tables 1 and 2 . ( b ) Recombinant Strep -hIntL-1 (50 µg/mL) binding to microbial glycan array. For glycan array data organized by genus, see Supplementary Fig. 2a . Data are presented as the mean ± s.d. ( n =4 technical replicates). The full data set can be found in Supplementary Table 3 . ( c ) Structural representation of the putative key binding epitopes for hIntL-1 and the non-binding N-acetylneuraminic acid (α-Neu5Ac). A terminal vicinal diol (red) is a common feature of α-Neu5Ac and all of the ligands identified.

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Recombinant, Binding Assay, Microarray

    Structure of hIntL-1 bound to allyl-β- d -Gal f . ( a ) Complex of hIntL-1 disulfide-linked trimer and allyl-β- d -Gal f . Each monomer unit is depicted in green, wheat, or grey, the β-allyl Gal f is shown in black, calcium ions in green, the inter-monomer disulfides in orange, and ordered water molecules in the binding site in red. The two orientations indicate the positioning of all three ligand-binding sites within the trimer. The trimeric structure is produced from Chain A in the asymmetric unit by a three-fold crystallographic operation. ( b ) Stereo image of the carbohydrate-binding site. Residues involved in calcium coordination and ligand binding are noted. Dashed lines are included to show the heptavalent coordination of the calcium ion and to highlight functional groups important for ligand and calcium ion binding. Difference density map (Fo-Fc, 3σ) of the allyl-β- d -Gal f ligand is provided in Supplementary Fig. 4b .

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: Structure of hIntL-1 bound to allyl-β- d -Gal f . ( a ) Complex of hIntL-1 disulfide-linked trimer and allyl-β- d -Gal f . Each monomer unit is depicted in green, wheat, or grey, the β-allyl Gal f is shown in black, calcium ions in green, the inter-monomer disulfides in orange, and ordered water molecules in the binding site in red. The two orientations indicate the positioning of all three ligand-binding sites within the trimer. The trimeric structure is produced from Chain A in the asymmetric unit by a three-fold crystallographic operation. ( b ) Stereo image of the carbohydrate-binding site. Residues involved in calcium coordination and ligand binding are noted. Dashed lines are included to show the heptavalent coordination of the calcium ion and to highlight functional groups important for ligand and calcium ion binding. Difference density map (Fo-Fc, 3σ) of the allyl-β- d -Gal f ligand is provided in Supplementary Fig. 4b .

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Binding Assay, Ligand Binding Assay, Produced, Functional Assay

    Models for hIntL-1 interacting with relevant saccharide epitopes from humans (α-Neu5Ac) or microbes (α-KDO). ( a ) Docking of methyl-α-Neu5Ac into the hIntL-1 structure. The conformation shown is similar to that observed in other protein structures with a methyl-α-Neu5Ac ligand (PDB: 2BAT, 2P3I, 2P3J, 2P3K, 2I2S, 1KQR, 1HGE, 1HGH (refs. 56 – 60 )). All models in this figure were generated from the allyl-β- d -Gal f –bound structure by docking the relevant diol of each compound into the Gal f diol electron density using Coot without further refinement. Calcium ions are shown in green and ordered water molecules are depicted in red. ( b ) Docking of methyl-α-KDO into the hIntL-1 structure. Comparison with methyl-α-Neu5Ac docked into the hIntL-1 structure reveal differences in the steric requirements for binding for each molecule.

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: Models for hIntL-1 interacting with relevant saccharide epitopes from humans (α-Neu5Ac) or microbes (α-KDO). ( a ) Docking of methyl-α-Neu5Ac into the hIntL-1 structure. The conformation shown is similar to that observed in other protein structures with a methyl-α-Neu5Ac ligand (PDB: 2BAT, 2P3I, 2P3J, 2P3K, 2I2S, 1KQR, 1HGE, 1HGH (refs. 56 – 60 )). All models in this figure were generated from the allyl-β- d -Gal f –bound structure by docking the relevant diol of each compound into the Gal f diol electron density using Coot without further refinement. Calcium ions are shown in green and ordered water molecules are depicted in red. ( b ) Docking of methyl-α-KDO into the hIntL-1 structure. Comparison with methyl-α-Neu5Ac docked into the hIntL-1 structure reveal differences in the steric requirements for binding for each molecule.

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Generated, Binding Assay

    Structures of the 20 most prevalent monosaccharides that are unique to bacterial glycans. The most common, l,d-α-heptose, is shown in the top left corner and number twenty, β- l -arabinose-4-N, is shown in the bottom right. This figure is derived from data in reference 32. Terminal acyclic 1,2-diol epitopes that could serve as ligands of hIntL-1 are highlighted with a red box.

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: Structures of the 20 most prevalent monosaccharides that are unique to bacterial glycans. The most common, l,d-α-heptose, is shown in the top left corner and number twenty, β- l -arabinose-4-N, is shown in the bottom right. This figure is derived from data in reference 32. Terminal acyclic 1,2-diol epitopes that could serve as ligands of hIntL-1 are highlighted with a red box.

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Derivative Assay

    hIntL-1 selectivity for monosaccharides. ( a ) Structures of saccharides used for characterization of hIntL-1 by ELISA and SPR. ( b ) The specificity of hIntL-1 for β-Gal f , β-ribofuranose (β-Rib f ) and β-galactopyranose (β-Gal p ) evaluated by ELISA (See Supplementary Fig. 1b for schematic). Data are presented as the mean ± s.d. ( n =3 technical replicates, data are representative of > 3 independent experiments). Data were fit to a single site binding equation (solid lines) and therefore represent the apparent (App) affinity of trimeric hIntL-1. Values for hIntL-1 bound to immobilized β-Gal f ( K d(App, Trimer) ± s.d.) are 85 ± 14 nM or 8.0 ± 1.3 µg/mL. ( c ) Representative real-time SPR sensorgrams of hIntL-1 binding to immobilized carbohydrates. Biotin served as a control. The SPR complete data set is available in Supplementary Fig. 1e .

    Journal: Nature structural & molecular biology

    Article Title: Recognition of Microbial Glycans by Human Intelectin

    doi: 10.1038/nsmb.3053

    Figure Lengend Snippet: hIntL-1 selectivity for monosaccharides. ( a ) Structures of saccharides used for characterization of hIntL-1 by ELISA and SPR. ( b ) The specificity of hIntL-1 for β-Gal f , β-ribofuranose (β-Rib f ) and β-galactopyranose (β-Gal p ) evaluated by ELISA (See Supplementary Fig. 1b for schematic). Data are presented as the mean ± s.d. ( n =3 technical replicates, data are representative of > 3 independent experiments). Data were fit to a single site binding equation (solid lines) and therefore represent the apparent (App) affinity of trimeric hIntL-1. Values for hIntL-1 bound to immobilized β-Gal f ( K d(App, Trimer) ± s.d.) are 85 ± 14 nM or 8.0 ± 1.3 µg/mL. ( c ) Representative real-time SPR sensorgrams of hIntL-1 binding to immobilized carbohydrates. Biotin served as a control. The SPR complete data set is available in Supplementary Fig. 1e .

    Article Snippet: hIntL-1 binding to Streptococcus pneumoniae Streptococcus pneumoniae (Klein) Chester serotype 8 (ATCC® 6308™), 20 (ATCC® 6320™), 43 (ATCC® 10343™) and 70 (ATCC® 10370™) were obtained from the ATCC.

    Techniques: Enzyme-linked Immunosorbent Assay, SPR Assay, Binding Assay

    Functional role of MASP-2 in pneumococcal meningitis mouse model. Kaplan-Meier curve showing increased survival in Masp2 −/− mice during pneumococcal meningitis ( a ). Cytokines and complement levels were measured in brain. Masp2 −/− mice had significantly lower brain levels of IL-1β, IL-10, and TNF-α 30 h after infection ( b – d ). Brain levels of C5b-9 were similar between Masp2 −/− and WT mice ( e ). Data are given as medians and 75th quartile; P values were determined with the Mann-Whitney U test

    Journal: Journal of Neuroinflammation

    Article Title: Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis

    doi: 10.1186/s12974-016-0770-9

    Figure Lengend Snippet: Functional role of MASP-2 in pneumococcal meningitis mouse model. Kaplan-Meier curve showing increased survival in Masp2 −/− mice during pneumococcal meningitis ( a ). Cytokines and complement levels were measured in brain. Masp2 −/− mice had significantly lower brain levels of IL-1β, IL-10, and TNF-α 30 h after infection ( b – d ). Brain levels of C5b-9 were similar between Masp2 −/− and WT mice ( e ). Data are given as medians and 75th quartile; P values were determined with the Mann-Whitney U test

    Article Snippet: Masp2 deficiency in experimental pneumococcal meningitis To investigate the role of MASP-2 in disease progression in pneumococcal meningitis, we infected Masp2 −/− (n = 12) and WT (n = 12) mice by intracisternal inoculation with S. pneumoniae serotype 3 (ATCC 6303) [ ].

    Techniques: Functional Assay, Mouse Assay, Infection, MANN-WHITNEY

    The effect of adjuvant treatment with the MASP-2 antibodies on clinical severity and survival in experimental pneumococcal meningitis. Kaplan-Meier curve of WT mice with pneumococcal meningitis treated intraperitoneally 20 h after infection with ceftriaxone (100 mg/kg) in combination with adjuvant treatment and observed for 68 h ( a ). Adjuvant treatment consisted of sterile saline, isotype antibodies (MAB205P, 1 mg/kg), or MASP-2 antibodies (D04211, 1 mg/kg). There was no difference in survival between groups. P values were determined with the log-rank test. Clinical severity scores for MASP-2 antibody-treated mice increased slower as compared to saline- (0.017 vs. 0.103 points/h) and isotype antibody (0.0017 vs. 0.080 points/h)-treated mice ( b ). P values were determined using linear mixed models with group/treatment, time, and their interaction as effects

    Journal: Journal of Neuroinflammation

    Article Title: Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis

    doi: 10.1186/s12974-016-0770-9

    Figure Lengend Snippet: The effect of adjuvant treatment with the MASP-2 antibodies on clinical severity and survival in experimental pneumococcal meningitis. Kaplan-Meier curve of WT mice with pneumococcal meningitis treated intraperitoneally 20 h after infection with ceftriaxone (100 mg/kg) in combination with adjuvant treatment and observed for 68 h ( a ). Adjuvant treatment consisted of sterile saline, isotype antibodies (MAB205P, 1 mg/kg), or MASP-2 antibodies (D04211, 1 mg/kg). There was no difference in survival between groups. P values were determined with the log-rank test. Clinical severity scores for MASP-2 antibody-treated mice increased slower as compared to saline- (0.017 vs. 0.103 points/h) and isotype antibody (0.0017 vs. 0.080 points/h)-treated mice ( b ). P values were determined using linear mixed models with group/treatment, time, and their interaction as effects

    Article Snippet: Masp2 deficiency in experimental pneumococcal meningitis To investigate the role of MASP-2 in disease progression in pneumococcal meningitis, we infected Masp2 −/− (n = 12) and WT (n = 12) mice by intracisternal inoculation with S. pneumoniae serotype 3 (ATCC 6303) [ ].

    Techniques: Mouse Assay, Infection

    Cerebrospinal fluid MASP-2 concentration and outcome in patients with pneumococcal meningitis. The MASP-2 concentration was significantly higher in patients with an unfavorable outcome ( n = 121) compared to that in patients with a favorable outcome ( n = 186). Each dot represents an individual patient, lines represent median values, and error bars are interquartile ranges. P value was determined with the Mann-Whitney U test

    Journal: Journal of Neuroinflammation

    Article Title: Mannose-binding lectin-associated serine protease 2 (MASP-2) contributes to poor disease outcome in humans and mice with pneumococcal meningitis

    doi: 10.1186/s12974-016-0770-9

    Figure Lengend Snippet: Cerebrospinal fluid MASP-2 concentration and outcome in patients with pneumococcal meningitis. The MASP-2 concentration was significantly higher in patients with an unfavorable outcome ( n = 121) compared to that in patients with a favorable outcome ( n = 186). Each dot represents an individual patient, lines represent median values, and error bars are interquartile ranges. P value was determined with the Mann-Whitney U test

    Article Snippet: Masp2 deficiency in experimental pneumococcal meningitis To investigate the role of MASP-2 in disease progression in pneumococcal meningitis, we infected Masp2 −/− (n = 12) and WT (n = 12) mice by intracisternal inoculation with S. pneumoniae serotype 3 (ATCC 6303) [ ].

    Techniques: Concentration Assay, MANN-WHITNEY

    mRNA expression of CD40L in PBMC cultures from 14 patients before and 4 to 6 weeks after immunization with 23-PV compared to mRNA expression in PBMC cultures from four unimmunized controls. Several patient values are very similar, so some lines are superimposed. Cells were stimulated for 5 h with the optimal concentration of pneumococcal polysaccharide serotype 3, 14 or 18C. ELISA-PCR results are expressed as copy numbers.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Up-Regulation of CD40 Ligand and Induction of a Th2 Response in Children Immunized with Pneumococcal Polysaccharide Vaccines

    doi: 10.1128/CDLI.8.2.233-240.2001

    Figure Lengend Snippet: mRNA expression of CD40L in PBMC cultures from 14 patients before and 4 to 6 weeks after immunization with 23-PV compared to mRNA expression in PBMC cultures from four unimmunized controls. Several patient values are very similar, so some lines are superimposed. Cells were stimulated for 5 h with the optimal concentration of pneumococcal polysaccharide serotype 3, 14 or 18C. ELISA-PCR results are expressed as copy numbers.

    Article Snippet: After an overnight incubation in RPMI 1640 at 37°C in humidified air containing 5% CO2 , PBMC (5 × 105 ) were cultured for 5 h in the absence or presence of concanavalin A (ConA) (20 mg/ml; Miles Scientific, Naperville Ill.), tetanus toxoid (TT) adsorbed USP (1:500; Connaught Laboratories Inc., Swiftwater, Pa.), 23-PV (1:500; Pnu-Immune, Lederle-Praxis Biologicals), and pneumococcal polysaccharide serotype 3, 14, or 18C (20 μg/ml; American Type Culture Collection, Manassas, Va.).

    Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction

    mRNA expression of IL-4 in PBMC cultures from 14 patients before and 4 to 6 weeks after immunization with 23-PV compared to mRNA expression in PBMC cultures from 4 unimmunized controls. Several patient values are very similar, so some lines are superimposed. Cells were stimulated for 5 h with the optimal concentration of pneumococcal polysaccharide serotype 3, 14, or 18C. ELISA-PCR results are expressed as copy numbers.

    Journal: Clinical and Diagnostic Laboratory Immunology

    Article Title: Up-Regulation of CD40 Ligand and Induction of a Th2 Response in Children Immunized with Pneumococcal Polysaccharide Vaccines

    doi: 10.1128/CDLI.8.2.233-240.2001

    Figure Lengend Snippet: mRNA expression of IL-4 in PBMC cultures from 14 patients before and 4 to 6 weeks after immunization with 23-PV compared to mRNA expression in PBMC cultures from 4 unimmunized controls. Several patient values are very similar, so some lines are superimposed. Cells were stimulated for 5 h with the optimal concentration of pneumococcal polysaccharide serotype 3, 14, or 18C. ELISA-PCR results are expressed as copy numbers.

    Article Snippet: After an overnight incubation in RPMI 1640 at 37°C in humidified air containing 5% CO2 , PBMC (5 × 105 ) were cultured for 5 h in the absence or presence of concanavalin A (ConA) (20 mg/ml; Miles Scientific, Naperville Ill.), tetanus toxoid (TT) adsorbed USP (1:500; Connaught Laboratories Inc., Swiftwater, Pa.), 23-PV (1:500; Pnu-Immune, Lederle-Praxis Biologicals), and pneumococcal polysaccharide serotype 3, 14, or 18C (20 μg/ml; American Type Culture Collection, Manassas, Va.).

    Techniques: Expressing, Concentration Assay, Enzyme-linked Immunosorbent Assay, Polymerase Chain Reaction