Journal: Journal of applied microbiology

Article Title: Simultaneous enrichment and optical detection of low levels of stressed Escherichia coli O157:H7 in food matrices.

doi: 10.1111/jam.12522

Figure Lengend Snippet: Fig. 1: Specificity of the E.coli O157:H7 biochip (a) Analysis of an E.coli O157:H7 culture (63 +/-17CFU/mL at the beginning of the experiment). (b) Analysis of an E.coli O157 non H7 culture (53+/-23 CFU/mL at the beginning of the experiment). O157:H7 (line with black circles) : anti-O157:H7 antibodies. O157 (line with empty circles) : anti-O157 antibodies. NC (solid line) : negative control antibodies.

Article Snippet: A purified monoclonal antibody against E. coli O157:H7 was obtained from Acris antibodies (Herford, Germany).

Techniques: Negative Control

Journal: Journal of applied microbiology

Article Title: Simultaneous enrichment and optical detection of low levels of stressed Escherichia coli O157:H7 in food matrices.

doi: 10.1111/jam.12522

Figure Lengend Snippet: Fig. 4: Detection of E.coli O157:H7 in complex food matrices.

Article Snippet: A purified monoclonal antibody against E. coli O157:H7 was obtained from Acris antibodies (Herford, Germany).

Techniques:

Journal: International Journal of Molecular Sciences

Article Title: Altered Intestinal Morphology and Microbiota Composition in the Autism Spectrum Disorders Associated SHANK3 Mouse Model

doi: 10.3390/ijms20092134

Figure Lengend Snippet: Altered gut morphology in Shank3αβ knock-out (KO) mice. ( A – D ) Histological evaluation of GI tract from wild type and Shank3αβ KO mice. ( A ) Longitudinal cross sections of Shank3αβ KO mice and wild type (WT) mice were stained with hematoxylin/eosin (HE) (upper panels) and periodic acid schiff (PAS) reaction (lower panels). Exemplary images are shown. ( B – D ) Morphological analysis of ( B ) villi length and ( C ) width, and ( D ) crypt depth reveals a significantly decreased villi length (Mann-Whitney U -test, p = 0.009; n = 5 animals per group) but not width ( p = 0.534), and normal crypt depth ( p = 0.983) in Shank3αβ KO mice. ( E , F ) Immunohistochemistry was performed on 5 mice per group and 5 optic fields of view each from 3 sections per mouse were analyzed. ( E ) A slight but non-significant decrease in FABP2 signal intensity was observed in Shank3αβ KO mice compared to wild types (left panel). Significantly higher ZONULIN-1 levels were found in Shank3αβ KO mice (right panel) ( t -test, p = 0.0413). ( F ) The levels of CLAUDIN3 and lipopolysaccharide (LPS) were not significantly different between Shank3αβ KO mice and wild types in gut epithelium. ( G ) Significantly higher ZONULIN-1 levels in Shank3αβ KO mice were confirmed by western blotting using gut epithelium protein lysate ( t -test, p = 0.0434, n = 3 per group). ( H ) Protein lysate from liver tissue from WT and Shank3αβ KO mice ( n = 3 per group) were analyzed for E. coli LPS levels using Western Blotting. The results show significantly higher LPS levels in the liver of Shank3αβ β KO mice ( t -test, p = 0.0452). * p < 0.05, ** p < 0.01.

Article Snippet: Zonulin 1 antibody was purchased from Thermo Fisher Scientific (Invitrogen) (Waltham, MA, USA); Claudin3 antibody from Abcam (Berlin, Germany); FABP2 antibody from Thermo Fisher Scientific (Invitrogen); LPS antibody from Origene (Rockville, MD, USA); and IL6 antibody was purchased from Cell signaling Technologies (Danvers, MA, USA); GFAP antibody was purchased from Sigma Aldrich (St. Louis, MO, USA); Cytokeratin and Vimentin antibody from Abcam.

Techniques: Knock-Out, Staining, MANN-WHITNEY, Immunohistochemistry, Western Blot

Journal: The Journal of Cell Biology

Article Title: Glypican and Biglycan in the Nuclei of Neurons and Glioma Cells: Presence of Functional Nuclear Localization Signals and Dynamic Changes in Glypican During the Cell Cycle

doi:

Figure Lengend Snippet: The location of potential nuclear localization signals in biglycan ( A ) and glypican ( B ) is indicated by the shaded regions, that were used for the construction of β-galactosidase fusion proteins. The clusters of basic amino acids are underlined. In one glypican mutant, the basic cluster was mutated to NSSSAN, and the sequence indicated by italics was deleted in a second mutant. The attachment sites for the phosphatidylinositol anchor and the COOH-terminal heparan sulfate chain of glypican are indicated by an open triangle and an arrow, respectively. In two fusion proteins, amino acids 499–558 of glypican were replaced by the human IgG Fc sequence (shown as hatched regions in C and D ), and in a deletion mutant, the potential nuclear localization signal used for the β-galactosidase fusion protein was removed from the glypican–Fc fusion protein ( D ).

Article Snippet: Monoclonal and rabbit polyclonal antibodies to Escherichia coli β-galactosidase were obtained from Promega (Madison, WI) and 5 Prime—3 Prime, Inc. (Boulder, CO), respectively, and were used for immunocytochemistry at a dilution of 1:500. mAbs to the HSV and human Fc tags were obtained from Novagen (Madison, WI) and Jackson ImmunoResearch Laboratories (West Grove, PA), respectively.

Techniques: Mutagenesis, Sequencing

Journal: The Journal of Cell Biology

Article Title: Glypican and Biglycan in the Nuclei of Neurons and Glioma Cells: Presence of Functional Nuclear Localization Signals and Dynamic Changes in Glypican During the Cell Cycle

doi:

Figure Lengend Snippet: Localization of β-galactosidase immunoreactivity in transfected 293 cells. 293 cells were transfected with β-galactosidase alone ( A and B ) or β-galactosidase fusion proteins containing either the putative nuclear localization signals of glypican ( C and D ) or biglycan ( E and F ). In a third fusion protein, the basic cluster of the glypican fragment was mutated to nonbasic amino acids ( G and H ), and in a fourth fusion protein ( I and J ), there was a further deletion of the amino acids shown in italics in Fig. B . A, C, E, G, and I show propidium iodide staining; B, D, F, H, and J show β-galactosidase immunoreactivity. Bars, 10 μm.

Article Snippet: Monoclonal and rabbit polyclonal antibodies to Escherichia coli β-galactosidase were obtained from Promega (Madison, WI) and 5 Prime—3 Prime, Inc. (Boulder, CO), respectively, and were used for immunocytochemistry at a dilution of 1:500. mAbs to the HSV and human Fc tags were obtained from Novagen (Madison, WI) and Jackson ImmunoResearch Laboratories (West Grove, PA), respectively.

Techniques: Transfection, Staining

Journal: PLoS ONE

Article Title: Membrane Insertion for the Detection of Lipopolysaccharides: Exploring the Dynamics of Amphiphile-in-Lipid Assays

doi: 10.1371/journal.pone.0156295

Figure Lengend Snippet: Using membrane insertion, two types of LPS were detected using their complement antibodies, polyclonal anti- E . coli O104 (pAb O104) and monoclonal anti- E . coli O111 (mAb O111) labeled with af647.

Article Snippet: Polyclonal antibody anti- E . coli O157 was from LifeSpan Biosciences (Seattle, WA). pAb E . coli O104, as well as monoclonal antibody for E . coli O111 were from Abraxis Inc. (Warminster, PA).

Techniques: Membrane, Labeling

Journal: PLoS ONE

Article Title: Membrane Insertion for the Detection of Lipopolysaccharides: Exploring the Dynamics of Amphiphile-in-Lipid Assays

doi: 10.1371/journal.pone.0156295

Figure Lengend Snippet: Schematics of (A) imaging flow cell and (B) waveguide flow cell. Major differences between these two flow cells include replacement of the waveguide glass substrate with a thin glass coverslip for imaging, and the addition of PDMS and silicone pedestals to create an airtight environment to preserve lipid integrity throughout an assay. (C) 100 μg/mL and (D) 50 μg/mL LPS O111:B4 incubated with BODIPY labeled DOPC lipids. Composite images of (E) 50 μg/mL and (F) 25 μg/mL LPS O157 bound by fluorescently labeled pAb O157-af647.

Article Snippet: Polyclonal antibody anti- E . coli O157 was from LifeSpan Biosciences (Seattle, WA). pAb E . coli O104, as well as monoclonal antibody for E . coli O111 were from Abraxis Inc. (Warminster, PA).

Techniques: Imaging, Incubation, Labeling

Journal: PLoS ONE

Article Title: Membrane Insertion for the Detection of Lipopolysaccharides: Exploring the Dynamics of Amphiphile-in-Lipid Assays

doi: 10.1371/journal.pone.0156295

Figure Lengend Snippet: (A) Bilayer prior to incubating with LPS O157. 50 μg/mL LPS (B) O157, (C) O104, and (D) O111:H11.

Article Snippet: Polyclonal antibody anti- E . coli O157 was from LifeSpan Biosciences (Seattle, WA). pAb E . coli O104, as well as monoclonal antibody for E . coli O111 were from Abraxis Inc. (Warminster, PA).

Techniques: