Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Results obtained following evaluation of the lytic activity of the isolated phage particles ph0031, ph0034, and ph0041, and a comparison with the lytic activity of phage JG004 obtained from the DSMZ collection ( a ) and of the lytic activity of the phage cocktail produced with the three isolated phage particles ( b ) on a P. aeruginosa DSM19880 bacterial lawn.

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Activity Assay, Isolation, Comparison, Produced

Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Results obtained in the optimization of the amounts of phage particles, sodium 1,2-naftoquinone-4-sulfonate, gelatin, and casein, leading to the bio-reactive chromogel for bio-detection system I, displaying the time evolution of the color produced using a fixed (added) amount of P. aeruginosa cells. The composition of formulations 1, 2, and 3 is displayed in .

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Produced

Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Normalized integrated color density of the three chromogels containing phage particles, sodium 1,2-naftoquinone-4-sulfonate, gelatin, and casein, further contacted with a fixed amount of P. aeruginosa cells, throughout the bioreaction time, allowing us to select an optimized bio-reactive chromogel. Values represent the mean of three independent assays. Error bars represent the standard deviation.

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Standard Deviation

Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Results obtained using the selected bio-reactive chromogel with optimized concentrations of phage particles, sodium 1,2-naftoquinone-4-sulfonate, gelatin, and casein, in bio-detection system II, with evolution throughout the time of the color produced using variable added amounts of P. aeruginosa cells. As a control for the colorimetric bioreaction, cells of S. aureus CCCD-S009 were also contacted with the selected bio-reactive chromogel ( , bottom right).

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Produced, Control

Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Normalized integrated color density of the optimized bio-reactive chromogel integrating phage particles, sodium 1,2-naftoquinone-4-sulfonate, gelatin, and casein, further added with variable amounts of P. aeruginosa cells, throughout the bioreaction time. Cells of S. aureus CCCD-S009 were used as a control for the bio-detection assays. Values represent the mean of three independent assays. Error bars represent the standard deviation.

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Control, Standard Deviation

Journal: Biosensors

Article Title: Bacteriophage-Based Biosensing of Pseudomonas aeruginosa : An Integrated Approach for the Putative Real-Time Detection of Multi-Drug-Resistant Strains

doi: 10.3390/bios11040124

Figure Lengend Snippet: Receptacle for the bio-reactive hydrogel in bio-detection system II, showing the position of the two light sensors ( a ), macroscopic aspect of the freshly-prepared phage-containing bio-reactive hydrogel ( b ), and macroscopic aspect of the phage-containing bio-reactive hydrogel with produced bioluminescence following exposure to P. aeruginosa cells ( c ).

Article Snippet: The collection strain of P. aeruginosa (ref. DSM 19880) used as a phage isolation host was acquired from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen GmmH, Braunschweig, Germany).

Techniques: Produced

Journal: WormBook : the online review of C. elegans biology

Article Title: Transgenic solutions for the germline *

doi: 10.1895/wormbook.1.148.1

Figure Lengend Snippet: Selected Entry Clones

Article Snippet: spe-11 promoter (no ATG) , in sperm , 272 bp spe-11 promoter with no start codon. Avoids attB1-encoded AAs in final fusion protein. ATG must be included in middle cassette (see ). , pCM1.128 (C.M. unpublished) , Addgene ( http://www.addgene.org/pgvec1 ).

Techniques: Clone Assay, Expressing, Sequencing

Journal: PLoS Pathogens

Article Title: DDX3 suppresses type I interferons and favors viral replication during Arenavirus infection

doi: 10.1371/journal.ppat.1007125

Figure Lengend Snippet: DDX3 ko-1, DDX3 ko-2 and WT A549 cells were infected with LCMV Cl13 (M.O.I. 0.5) for the indicated times. Total RNA in cell lysates was extracted and normalized interferon transcript levels ( ifnb/gapdh) determined by qRT-PCR and represented as relative fold expression. B. DDX3 ko-1 and WT A549 cells were transduced with empty-RV (EV-RV) or RV encoding DDX3 (DDX3-RV) before infection, and processed as in A for quantification of ifnb/gapdh transcripts via qRT-PCR ( B, left panel) or determination of bioactive IFN-I levels in cell culture supernatants at indicated h.p.i. ( B, right panel). C. DDX3 ko-1 and WT A549 cells were pre-incubated for 2 h and infected with LCMV Cl13 (M.O.I. 0.5) in the presence of anti-IFNAR mAb (IFNAR Ab, solid lines) or Isotype control (Iso Ab, broken lines), which were left for the remaining of the culture. Viral titers were determined at indicated h.p.i. D . Translation assay performed in HEK-293T cells treated with DDX3 or scrambled (Scr) siRNA. E. Minireplicon assay performed in DDX3 ko-1 or WT A549 cells. F and G. DDX3 ko-1 or WT cells were transfected with 0.4 μg of empty plasmid, plasmid expressing DDX3 ( F-G ) or the indicated point-mutant DDX3 ( G ) and used for minireplicon assay. 100% value was given to WT A549 transfected with empty plasmid. Data are representative of 2 ( A-C & E-F ) or 3 ( D & G ) independent experiments. * p<0.05, ** p<0.01, ***p<0.001. Stars represent: DDX3ko vs WT (blue) or DDX3 ko at the indicated h.p.i. versus DDX3 ko at time = 0 (red) ( A ), DDX3 ko-EV vs WT-EV (black) or vs DDX3 ko-RV-DDX3 (red) ( B ), DDX3 ko-IFNAR vs WT-IFNAR (black) or vs DDX3 ko-Isotype (red) ( C ).

Article Snippet: Anti-IFNAR antibody (#21385–1, PBL Interferon Source) and Isotype control IgG2a antibody (#554126, BD Pharmingen Product) were both used at 5 μg/ml.

Techniques: Infection, Quantitative RT-PCR, Expressing, Transduction, Cell Culture, Incubation, Control, Transfection, Plasmid Preparation, Mutagenesis

Journal: PLoS ONE

Article Title: Chikungunya virus superinfection exclusion is mediated by a block in viral replication and does not rely on non-structural protein 2

doi: 10.1371/journal.pone.0241592

Figure Lengend Snippet: ( A – C ) HFFs were treated with human IFNAR blocking antibody or isotype control at 5 μg/mL for 1 h, then were infected with CHIKV-GFP for 24 h at the indicated MOI, followed by CHIKV-mCherry at MOI 1 for 24 h. Blocking antibody treatment was maintained throughout the experiment. Representative flow cytometry plots (A) and quantification of infected cells (C) and are shown. Arrows highlight the difference in GFP infection between isotype control and blocking antibody treated samples. MX1 RNA levels were assessed by RT–qPCR (B). ( D , E ) WT (D) or Irf3 −/− Irf7 −/− MEF cells (E) were infected with CHIKV-GFP at the indicated MOI for 16 h, then with CHIKV-mCherry at MOI 5 (WT) or 3 ( Irf3 −/− Irf7 −/− ) for 8 h, and subsequently analyzed by flow cytometry. Bars indicate mean and SD of biological triplicates, and data are representative of at least two independent experiments. NS, not significant; *** p < 0.001 (one-way analysis of variance followed by Dunnett’s post-test).

Article Snippet: Human IFNAR blocking antibody (PBL Assay Science #21385–1) was used at a concentration of 5 μg/mL, ActD (Sigma-Aldrich #A1410) at 2 μg/μL.

Techniques: Blocking Assay, Control, Infection, Flow Cytometry, Quantitative RT-PCR

Journal: Cell reports

Article Title: Constitutive Interferon Attenuates RIPK1/3-Mediated Cytokine Translation

doi: 10.1016/j.celrep.2019.12.073

Figure Lengend Snippet: (A) TNF-α and CXCL-1 protein levels after indicated stimulations ± treatment with eIF4E inhibitor 4EGi-1 in B6, Ifnb −/− , and RIP1 Ki Ifnb −/− BMDMs. (B) p-eIF4E and total 4E-BP levels in B6, Ifnb −/− , and RIP1 Ki BMDMs stimulated as indicated ± 5 IU IFNβ priming overnight. (C) TNF-α and CXCL-1 protein levels after indicated stimulations ± treatment with mTORC1 inhibitor Torin 2 in B6, Ifnb −/− , and RIP1 Ki Ifnb −/− BMDMs. (D and E) Cell death as measured by propidium iodide incorporation over 6 h in B6, Ifnb −/− , and RIP1 Ki Ifnb −/− BMDMs stimulated with LZ ± treatment with 4EGi-1 (D) or Torin 2 (E). (F) TNF-α and CXCL-1 protein levels after indicated stimulations ± overnight treatment with IFNAR-blocking antibody (αIFNAR) in Akt1 −/− and wild-type (WT) littermate BMDMs. In all panels, BMDMs were stimulated with LPS, LZ, LZNs, Z, Ns, or LZN as indicated. ELISA and qPCR data are shown as ±SD from three independent experiments compared using two-way ANOVA: n.s. (p > 0.05), *p < 0.05, and ****p < 0.0001. Western blot and kinetic cell death experiments are representative of three or more independent experiments. Kinetic data are presented as the mean ± SD of triplicate wells. See also and .

Article Snippet: Blocking antibody to Human IFNAR (21385–1) was purchased from PBL Assay Science and used at 20ug/ml.

Techniques: Blocking Assay, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Cell reports

Article Title: Constitutive Interferon Attenuates RIPK1/3-Mediated Cytokine Translation

doi: 10.1016/j.celrep.2019.12.073

Figure Lengend Snippet: (A) Cell death as measured by propidium iodide incorporation over 8 h in human PBMC-derived macrophages stimulated as indicated ± overnight treatment with IFNAR-blocking antibody. (B and C) CXCL-1 protein (B) and mRNA (C) levels in human PBMC-derived macrophages stimulated as indicated ± overnight treatment with IFNAR-blocking antibody (D and E) CXCL-1 protein (D) and mRNA (E) levels in human PBMC-derived macrophages stimulated as indicated ± overnight treatment with IFNAR-blocking antibody and/or treatment with 4EGi-1 or Torin 2. In all panels, human PBMC-derived macrophages were stimulated with LPS, LZ, or LZNs as indicated. ELISA and qPCR data are shown as ±SD from three independent experiments compared using two-way ANOVA: n.s. (p > 0.05), **p < 0.01, ***p < 0.001, and ****p < 0.0001. Kinetic cell death experiments are representative of three or more independent experiments and presented as the mean ± SD of triplicate wells.

Article Snippet: Blocking antibody to Human IFNAR (21385–1) was purchased from PBL Assay Science and used at 20ug/ml.

Techniques: Derivative Assay, Blocking Assay, Enzyme-linked Immunosorbent Assay

Journal: Cell reports

Article Title: Constitutive Interferon Attenuates RIPK1/3-Mediated Cytokine Translation

doi: 10.1016/j.celrep.2019.12.073

Figure Lengend Snippet:

Article Snippet: Blocking antibody to Human IFNAR (21385–1) was purchased from PBL Assay Science and used at 20ug/ml.

Techniques: Purification, Recombinant, Enzyme-linked Immunosorbent Assay, RNA Sequencing, Derivative Assay