s pneumoniae  (ATCC)

Journal: Critical Care

Article Title: Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats

doi: 10.1186/cc8040

Figure Lengend Snippet: Pulmonary coagulation and fibrinolysis. The effects of anticoagulants nebulized into the lungs of rats on levels of (a) thrombin-antithrombin complexes (TATc), (b) antithrombin activity (AT), (c) fibrin degradation products (FDP), (d) plasminogen activator activity (PAA%), and (e) plasminogen activator inhibitor-1 (PAI-1) in bronchoalveolar lavage fluid (BALF), 40 hours after intra-tracheal bacterial challenge ( Streptococcus pneumoniae , serotype 3, ATCC 6303). Dotted lines stipulate the normal values in healthy animals and untreated animals with pneumonia. Data represent mean ± standard deviation. * P < 0.01 vs. placebo. APC = recombinant-human activated protein C; AT = plasma = derived human antithrombin; Hep = heparin; Dan = danaparoid.

Article Snippet: Pneumonia was induced in male Sprague-Dawley rats (weighing 250 to 300 g; Harlan, Horst, The Netherlands) by intratracheal instillation of 5 × 10 6 colony-forming units (CFU) of S. pneumoniae (serotype 3, ATCC 6303) as described previously [ ].

Techniques: Coagulation, Activity Assay, Standard Deviation, Recombinant, Derivative Assay

Journal: Critical Care

Article Title: Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats

doi: 10.1186/cc8040

Figure Lengend Snippet: The effects of anticoagulants on numbers of Streptococcus pneumoniae colony forming units in (a) lung homogenate and (b) bronchoalveolar lavage fluid (BALF), 40 hours after intra-tracheal bacterial challenge. Data represent median ± interquartile range. * P < 0.05 vs. placebo. APC = recombinant-human activated protein C; AT = plasma = derived human antithrombin; CFU = colony-forming units; Hep = heparin; Dan = danaparoid.

Article Snippet: Pneumonia was induced in male Sprague-Dawley rats (weighing 250 to 300 g; Harlan, Horst, The Netherlands) by intratracheal instillation of 5 × 10 6 colony-forming units (CFU) of S. pneumoniae (serotype 3, ATCC 6303) as described previously [ ].

Techniques: Recombinant, Derivative Assay

Journal: Critical Care

Article Title: Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats

doi: 10.1186/cc8040

Figure Lengend Snippet: Total cell and neutrophil counts in bronchoalveolar lavage fluid

Article Snippet: Pneumonia was induced in male Sprague-Dawley rats (weighing 250 to 300 g; Harlan, Horst, The Netherlands) by intratracheal instillation of 5 × 10 6 colony-forming units (CFU) of S. pneumoniae (serotype 3, ATCC 6303) as described previously [ ].

Techniques:

Journal: Critical Care

Article Title: Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats

doi: 10.1186/cc8040

Figure Lengend Snippet: Histopathological changes in Streptococcus pneumoniae pneumonia. Shown are representative hematoxylin and eosin-stained photomicrographs (magnification, ×100) of lung tissue from rats treated with (a) saline, (b) recombinant human activated protein C (APC), (c) plasma-derived antithrombin (AT), (d) heparin (Hep), (e) danaparoid (Dan), at 40 hours after bacterial challenge. (f) Total histopathologic scores are presented as median with interquartile range. * P < 0.05 vs. placebo.

Article Snippet: Pneumonia was induced in male Sprague-Dawley rats (weighing 250 to 300 g; Harlan, Horst, The Netherlands) by intratracheal instillation of 5 × 10 6 colony-forming units (CFU) of S. pneumoniae (serotype 3, ATCC 6303) as described previously [ ].

Techniques: Staining, Recombinant, Derivative Assay

Journal: Critical Care

Article Title: Nebulized antithrombin limits bacterial outgrowth and lung injury in Streptococcus pneumoniae pneumonia in rats

doi: 10.1186/cc8040

Figure Lengend Snippet: The effects of BALF from placebo treated rats (BALF-none) and BALF from rats treated with plasma-derived AT (BALF-AT) on survival of Streptococcus pneumoniae in vitro . (a) After five hours outgrowth of S. pneumoniae in bronchoalveolar lavage fluid from rats treated with plasma-derived antithrombin (BALF-AT) was reduced (closed symbols), compared with the outgrowth in BALF-none samples (open symbols). * P < 0.0001 vs. BALF-AT. (b) In vitro added plasma-derived AT did not affect the number of colony-forming units (CFU; for clarity only the highest concentrations are shown: 1.1 mg/mL, open symbols; 4.4 mg/mL, closed symbols). (c) Sodium polyanetholsulphonate (SPS) blocked the inhibitory effect of BALF-AT on outgrowth of S. pneumoniae (BALF-AT without SPS, closed symbols; BALF-AT with SPS, open symbols). Data represent mean ± standard deviation. * P < 0.0001 vs. BALF-AT without SPS.

Article Snippet: Pneumonia was induced in male Sprague-Dawley rats (weighing 250 to 300 g; Harlan, Horst, The Netherlands) by intratracheal instillation of 5 × 10 6 colony-forming units (CFU) of S. pneumoniae (serotype 3, ATCC 6303) as described previously [ ].

Techniques: Derivative Assay, In Vitro, Standard Deviation

Journal: BMC Infectious Diseases

Article Title: Inflammasome activation mediates inflammation and outcome in humans and mice with pneumococcal meningitis

doi: 10.1186/1471-2334-13-358

Figure Lengend Snippet: Expression of NLRP3 and ASC in brain homogenates. Expression of NLRP3/GAPDH (A) and ASC/GAPDH (B) in brain homogenates of WT, Asc −/− and Nlrp3 −/− mice 6 hours and 30 hours after induction of S . pneumoniae meningitis compared to mice inoculated with saline. Three mice per group were analysed in the sham infected mice and the 6 hour timepoint; 4 mice were analyzed at the 30 hour timepoint. Data are given as means +/− SD.

Article Snippet: Serotype 3 S . pneumoniae (ATCC 6303; American Type Culture Collection, Rockville, MD, USA) was grown to mid log phase in 4 hours at 37°C in Todd-Hewitt broth supplemented with yeast (Difco, Detroit, MI).

Techniques: Expressing, Infection

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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 < 0.001; *, p < 0.05) compared to control as determined by ANOVA followed by the Bonferroni test.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

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

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

Techniques: Expressing, Infection, Immunolabeling, Fluorescence, Microscopy

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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 < 0.05; **, p < 0.01) indicates a significant difference between Camp expression after treatment and control (as determined by ANOVA followed by the Bonferroni test). Astrocytes or microglia were incubated with NM (C) or SP (D) with or without 1, 5 or 10 μM WRW4 (30 min preincubation) and WRW4 alone for 24 h or 12 h, respectively. Glial cells were fixed and labelled with anti-rCRAMP antibodies and protein expression was examined by immunofluorescence microscopy. Bisbenzimide was used for nuclear counter-staining (blue). The figures show representative results from one of three independent experiments. Scale bar: 20 μm.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

Techniques: Inhibition, Expressing, Incubation, Immunofluorescence, Microscopy, Staining

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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 < 0.05; **, p < 0,01; ***, p < 0,001) compared to controls (also with DMSO in equivalent amount) as determined by one-way ANOVA and Bonferroni post-hoc tests. In order to analyse the inhibition of forskolin-stimulated adenylate cyclase activity, astrocytes (E) and microglia (F) were subjected to either 10 μM (E) or 25 μM (F) forskolin as well as to NM, SP or 1 μM fMLF and NM, SP or fMLF with 10 μM WRW4 (30 min preincubation) and to WRW4 alone for 15 min at 37°C. cAMP levels were determined as described above (see Methods). The values given represent mean ± SEM from four independent experiments. Asterisks indicate significant differences (*, p < 0.05; ***, p < 0,001) between forskolin plus agonists and forskolin alone as determined by one-way ANOVA and Bonferroni post-hoc tests.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

Techniques: Inhibition, Western Blot, Derivative Assay, Activity Assay

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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 < 0.05; **, p < 0,01; ***, p < 0,001) compared to control (one-way ANOVA followed by the Bonferroni test). For analysis of inhibition of forskolin-stimulated adenylate cyclase activity, untransfected (I) or hMARCO- (J) , hFPRL1- (K) , and hFPRL1/hMARCO- (L) expressing HEK293 cells were subjected to 25 μM forskolin as well as to NM or 1 μM fMLF for 15 min at 37°C. cAMP levels were determined as described above (see Methods). The values represent mean ± SEM from four independent experiments. Asterisks indicate a significant difference (*, p < 0.05; **, p < 0,01) between forskolin plus agonists and forskolin alone, as determined via one-way ANOVA followed by the Bonferroni test.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

Techniques: Transfection, Expressing, Western Blot, Inhibition, Activity Assay

Journal: Journal of Neuroinflammation

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

doi: 10.1186/1742-2094-8-11

Figure Lengend 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.

Article Snippet: Cells were challenged with supernatants from Neisseria meningitidis (ATCC 13077, type strain isolated from a fatal case of meningitis; 1:100) or Streptococcus pneumoniae (ATCC 6303; capsula type 3; 1:100) in serum and antibiotic-free DMEM.

Techniques: Expressing, Transfection, Quantitative RT-PCR

Journal: International Journal of Nanomedicine

Article Title: Tat-functionalized liposomes for the treatment of meningitis: an in vitro study

doi: 10.2147/IJN.S130125

Figure Lengend Snippet: Streptococcus pneumoniae treated with Van, Tat-LipoVan, and LipoVan. Notes: Antibiotic concentrations of ( A ) 0.2 µg/mL, ( B ) 0.3 µg/mL, ( C ) 0.7 µg/mL, and ( D ) 1 µg/mL. Data presented as means (n=3). Abbreviations: Van, vancomycin; Tat-LipoVan, Tat-functionalized Van-loaded liposomes; OD, optical density.

Article Snippet: S. pneumoniae (Klein) Chester (6303), multidrug-resistant E. coli (25922), and MRSA (43360) where purchased from ATCC.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Tat-functionalized liposomes for the treatment of meningitis: an in vitro study

doi: 10.2147/IJN.S130125

Figure Lengend Snippet: Colony-forming units (CFU) of Streptococcus pneumoniae after treatment with free Van, LipoVan, and Tat-LipoVan for 8 hours. Notes: * P <0.05 versus free Van and LipoVan (0.2 µg/mL); ** P <0.05 versus free Van (0.6 µg/mL); *** P <0.05 versus free Van (1 µg/mL). Abbreviations: Van, vancomycin; Tat-LipoVan, Tat-functionalized Van-loaded lipo somes.

Article Snippet: S. pneumoniae (Klein) Chester (6303), multidrug-resistant E. coli (25922), and MRSA (43360) where purchased from ATCC.

Techniques:

Journal: International Journal of Nanomedicine

Article Title: Tat-functionalized liposomes for the treatment of meningitis: an in vitro study

doi: 10.2147/IJN.S130125

Figure Lengend Snippet: Live/dead assays with Streptococcus pneumoniae and methicillin-resistant Staphylococcus aureus (MRSA). Notes: Live bacteria in green, and dead bacteria in red. Magnification 10×. Antibiotic concentrations of 0.6 µg/mL for vancomycin and 2 µg/mL for methicillin.

Article Snippet: S. pneumoniae (Klein) Chester (6303), multidrug-resistant E. coli (25922), and MRSA (43360) where purchased from ATCC.

Techniques: