Journal: Microorganisms

Article Title: Shiga-Toxin-Producing Strains of Escherichia coli O104:H4 and a Strain of O157:H7, Which Can Cause Human Hemolytic Uremic Syndrome, Differ in Biofilm Formation in the Presence of CO 2 and in Their Ability to Grow in a Novel Cell Culture Medium

doi: 10.3390/microorganisms11071744

Figure Lengend Snippet: E. coli strains used in the present study.

Article Snippet: B shows the color of the E. coli cultures in the HBD cell culture medium after 48 h. The E. coli O104:H4 Ec 01, Ec 02, and Ec 03 cultures were green, while the E. coli ATCC 25922 and O157:H7 cultures remained pink (unchanged).

Techniques:

Journal: Microorganisms

Article Title: Shiga-Toxin-Producing Strains of Escherichia coli O104:H4 and a Strain of O157:H7, Which Can Cause Human Hemolytic Uremic Syndrome, Differ in Biofilm Formation in the Presence of CO 2 and in Their Ability to Grow in a Novel Cell Culture Medium

doi: 10.3390/microorganisms11071744

Figure Lengend Snippet: CO 2 enhances biofilm formation by the E. coli O104:H4 strains but not the E. coli O157:H7 strains. The E. coli strains that were examined were E. coli O104:H4 (strains Ec01, Ec02, and Ec03), ATCC 25922, and O157:H7. The cultures were incubated in MH II broth overnight at 37 °C. All strains were examined in triplicate, and figures represent 1 of 3 separate studies. Only a single well of the triplicate is shown ( A , B ). A. Biofilm formation by the E. coli strains in a 96-well plate with and without CO 2 . B. Biofilm formation by the E. coli strains in 5 mL culture tubes with and without CO 2 . ( C ) Quantitative crystal violet analysis of the amount of biofilm formed by the E. coli strains in 5 mL culture tubes with and without CO 2 . In ( C ), results are presented as geometric mean with standard error of the mean. p -values are shown for the comparison of the amount of biofilm formation by E. coli O157:H7 with (orange line) and without (blue line) CO 2 and the amount of biofilm formation by the O104:H4 strains under the same conditions. p -values, when significant, are also shown for comparison of biofilm formation between individual strains with and without CO 2 .

Article Snippet: B shows the color of the E. coli cultures in the HBD cell culture medium after 48 h. The E. coli O104:H4 Ec 01, Ec 02, and Ec 03 cultures were green, while the E. coli ATCC 25922 and O157:H7 cultures remained pink (unchanged).

Techniques: Incubation, Comparison

Journal: Microorganisms

Article Title: Shiga-Toxin-Producing Strains of Escherichia coli O104:H4 and a Strain of O157:H7, Which Can Cause Human Hemolytic Uremic Syndrome, Differ in Biofilm Formation in the Presence of CO 2 and in Their Ability to Grow in a Novel Cell Culture Medium

doi: 10.3390/microorganisms11071744

Figure Lengend Snippet: E. coli O104:H4 strains appear to grow in a novel cell culture medium but the E. coli O157:H7 strain does not. Live E. coli strains were examined for their ability to activate TLR2 or 4. The figure presents representative results from one of three studies we conducted with live bacterial strains after overnight incubation at 37 °C. ( A ) Bacterial strains and TLR control antigens were added in triplicate wells of HEK294 cells expressing TLR2 or 4 in an HBD cell culture medium overnight at 37 °C with 5% CO 2 in 96-well plates. Note the color reaction difference between the E. coli O104:H4 strains and the ATCC 25922 and O157:H7 strains. ( B ) The figure shows the amount of TLR2 or 4 activation by live E. coli strains. The results are presented as geometric means with standard errors of the mean. p -values are also shown comparing the amount of TLR2 activated by E. coli O157:H7 to the amount of TLR2 activated by the other E. coli strains (in blue). The differences between the results of TLR4 activation by E. coli O157:H7 and the other strains were not significant. Statistical comparisons are also shown between the amount of TLR2 and 4 activation of the individual E. coli strains. ( C ) Reaction using live E. coli strains with HEK293 cells not expressing TLR2 or 4 (null cells). ( D ) Incubation of live E. coli strains in HBD cell culture medium or DMEM cell culture maintenance medium without HEK293 cells.

Article Snippet: B shows the color of the E. coli cultures in the HBD cell culture medium after 48 h. The E. coli O104:H4 Ec 01, Ec 02, and Ec 03 cultures were green, while the E. coli ATCC 25922 and O157:H7 cultures remained pink (unchanged).

Techniques: Cell Culture, Incubation, Control, Expressing, Activation Assay

Journal: Microorganisms

Article Title: Shiga-Toxin-Producing Strains of Escherichia coli O104:H4 and a Strain of O157:H7, Which Can Cause Human Hemolytic Uremic Syndrome, Differ in Biofilm Formation in the Presence of CO 2 and in Their Ability to Grow in a Novel Cell Culture Medium

doi: 10.3390/microorganisms11071744

Figure Lengend Snippet: Differential growth of E. coli O104:H4 and O157:H7 strains in HBD cell culture medium. Single colonies of the E. coli O104:H4 or O157:H7 (and ATCC 25922) strains from SBA plates were used to inoculate 2 mL of MH II broth and cultures were incubated overnight at 37 °C. Overnight E. coli cultures were used to inoculate 12.5 mL of HBD cell culture medium (0.25 mL inoculum) or 12 mL MH II broth (0.12 mL inoculum) in 125 mL flasks and cultures were incubated at 37 °C with shaking at 180 rpm. Growth was followed by an increase in absorbance at 600 nm. The figure presents representative results from one of three independent studies. ( A ) Growth of the E . coli strains in HBD cell culture medium followed for 48 h. ( B ) Appearance of the E. coli cultures in HBD cell culture medium after 48 h. ( C ) Growth of the E. coli strains in MH II broth after 7 h of incubation.

Article Snippet: B shows the color of the E. coli cultures in the HBD cell culture medium after 48 h. The E. coli O104:H4 Ec 01, Ec 02, and Ec 03 cultures were green, while the E. coli ATCC 25922 and O157:H7 cultures remained pink (unchanged).

Techniques: Cell Culture, Incubation

Journal: Microorganisms

Article Title: Shiga-Toxin-Producing Strains of Escherichia coli O104:H4 and a Strain of O157:H7, Which Can Cause Human Hemolytic Uremic Syndrome, Differ in Biofilm Formation in the Presence of CO 2 and in Their Ability to Grow in a Novel Cell Culture Medium

doi: 10.3390/microorganisms11071744

Figure Lengend Snippet: Growth of E. coli O104:H4 strains in HBD cell culture medium starting with single colonies. A single colony from an overnight growth on SBA plates was used to inoculate 2 mL of HBD medium and cultures were incubated overnight at 37 °C with 5% CO 2 . The figure presents representative results from one of three independent studies. There were color differences between the E. coli O104:H4 strains and the ATCC 25922 and O157:H7 strains. This, along with the pH differences (pH ~5.0 vs. 8.5–9.0), reflected the growth of the E. coli O104:H4 strains. The purple color of the E. coli ATCC and O157:H7 cultures may have been derived from endogenous alkaline phosphatase that came from the original inoculum.

Article Snippet: B shows the color of the E. coli cultures in the HBD cell culture medium after 48 h. The E. coli O104:H4 Ec 01, Ec 02, and Ec 03 cultures were green, while the E. coli ATCC 25922 and O157:H7 cultures remained pink (unchanged).

Techniques: Cell Culture, Incubation, Derivative Assay

Journal: IDCases

Article Title: An Unusual Carbapenem Resistant Escherichia coli Carrying Plasmid-mediated AmpC and Mutated ompC in A Patient with Recurrent Urinary Tract Infections

doi: 10.1016/j.idcr.2020.e00781

Figure Lengend Snippet: Antimicrobial Susceptibility Profile of EC01. Broth microdilution was used to determine susceptibility of EC01 to the antibiotics. The interpretive criteria were based on CLSI 2019 breakpoints. Cefazolin was used to predict susceptibility to oral cephalosporins.

Article Snippet: The susceptibility of the E. coli isolate (designated EC01) was performed by broth microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) reference using panels prepared in house.

Techniques:

Journal: IDCases

Article Title: An Unusual Carbapenem Resistant Escherichia coli Carrying Plasmid-mediated AmpC and Mutated ompC in A Patient with Recurrent Urinary Tract Infections

doi: 10.1016/j.idcr.2020.e00781

Figure Lengend Snippet: EC01 is most closely related to E. coli strain D7. A Kmer tree was constructed using the most closely related strains as well as a large sampling of E. coli strains from around the world and derived from different sources.

Article Snippet: The susceptibility of the E. coli isolate (designated EC01) was performed by broth microdilution method according to the Clinical and Laboratory Standards Institute (CLSI) reference using panels prepared in house.

Techniques: Construct, Sampling, Derivative Assay

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and culture of adult murine cardiac atrial and ventricular fibroblasts and myofibroblasts.

doi: 10.1016/j.ymeth.2021.04.004

Figure Lengend Snippet: Fig. 3. Phase contrast images of P0 atrial and ventricular fibroblast and myofibroblast monolayer at different time points. Cardiac fibroblasts isolated from mouse and rat heart (ventricles and atria) were seeded onto ‘non-compliant’ uncoated standard polystyrene cell culture dishes with an elastic modulus of ≈107 kPa or ‘compliant’ collagen-coated hydrogel-bound polystyrene plates with a much lower elastic modulus of 8 kPa. (A) Mouse ventricular fibroblasts (8 kPa) and myofi broblasts (107 kPa) imaged at the indicated time after plating. (B) Mouse atrial fibroblasts (8 kPa) and myofibroblasts (107 kPa) imaged at the indicated time after plating. (C) Rat ventricular myofibroblasts (107 kPa) imaged 150 min (0 h) after isolation. (D) Rat atrial myofibroblasts (107 kPa) imaged 150 min (0 h) after isolation. Scale bar = 50 μm.

Article Snippet: For immunocytofluorescence assays, cells were seeded onto either normal glass coverslips in standard polystyrene culture dishes (noncompliant substrate) or collagen-coated soft hydrogel-bound glass coverslips (Matrigen #SS6-COL-8) inserted into compliant culture dishes from Matrigen.

Techniques: Isolation, Cell Culture

Journal: Methods (San Diego, Calif.)

Article Title: Isolation and culture of adult murine cardiac atrial and ventricular fibroblasts and myofibroblasts.

doi: 10.1016/j.ymeth.2021.04.004

Figure Lengend Snippet: Fig. 6. The abundance and cytoskeletal incorporation of smooth muscle α-actin immunoreactivity are attenuated when cardiac fibroblasts are cultured on a compliant substrate. (A) Cardiac fibroblasts isolated from mouse ventricles were seeded onto ‘non-compliant’ uncoated standard cell culture dishes (elastic modulus 107 kPa) or ‘compliant’ collagen-coated hydrogel-bound polystyrene plates (elastic modulus 8 kPa). Lysates (20 μg/lane) from passage 2 cells were used for immunoblot assays. A representative immunoblot showing the abundance of smooth muscle α-actin (αSMA) immunoreactivity from 1 of 3 independent experiments is shown on the left. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was used as a loading control. Numbers on the right of the immunoblot indicate the position of molecular mass markers (in kDa). The histogram on the right shows the mean ± SEM of three independent experiments. (B) Cardiac fibroblasts were seeded onto either ‘non-compliant’ uncoated glass coverslips in standard polystyrene culture dishs (elastic modulus 107 kPa) or ‘compliant’ collagen-coated soft hydrogel-bound glass coverslips (Matrigen #SS6-COL-8) inserted into compliant culture dishes from Matrigen (elastic modulus 8 kPa). Passage 2 cells were fixed with 2% paraformaldehyde, permeabilized, and decorated with Alexa Fluor 555-phalloidin (to identify actin filaments) and antibodies to αSMA. The images were acquired using a Zeiss LSM-510 confocal fluorescence microscope. Nuclei were visualized using the DNA-binding fluorescent dye, DAPI, and overlay images of the staining are shown (Merge). Images are representative of three independent biological replicates. Scale bar = 20 μm.

Article Snippet: For immunocytofluorescence assays, cells were seeded onto either normal glass coverslips in standard polystyrene culture dishes (noncompliant substrate) or collagen-coated soft hydrogel-bound glass coverslips (Matrigen #SS6-COL-8) inserted into compliant culture dishes from Matrigen.

Techniques: Cell Culture, Isolation, Western Blot, Control, Fluorescence, Microscopy, Binding Assay, Staining