Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: Homologies of gene rpoD among Staphylococcus species.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques:

Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: Anti- rpoD PPNA2332 was added to cell cultures containing 1.0×10 5 CFU/mL MRSA/VISA Mu50 to a final concentration of 6.25, 12.5, or 25 M. Additional cell cultures were treated with free MH broth, scrambled PPNA2332 (final concentration of 40 µM), and peptide (KFF) 3 K (final concentration of 40 µM) in a volume equal to that of the PPNA2332 preparation as controls. The growth of different groups of MRSA/VISA Mu50 cells was monitored by using OD measurements. The data are shown as means for 2 samples from 2 independent tests.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques: Concentration Assay

Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: Anti- rpoD PPNA2332 was added to cell cultures containing 1.0×10 5 CFU/mL MRSA/VISA Mu50 to a final concentration of 5, 10, 20, or 40 µM. Additional cell cultures were treated with free MH broth, scrambled PPNA2332 (final concentration of 40 µM), and peptide (KFF) 3 K (final concentration of 40 µM) in a volume equal to that of the PPNA2332 preparation as controls. Aliquots of each culture were collected at 0 h and 24 h, diluted, and inoculated onto solid MH agar. The number of CFU was calculated from the number of colonies growing on plates. The 24 h counts for 40 µM PPNA2332 were 0 CFUs/mL. The data are shown as means ± SD for 2 samples from 2 independent test. *, P<0.01 for comparison to control values.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques: Concentration Assay, Comparison, Control

Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: Triplicate bacterial cultures were grown for 18 h in 100 µL of MH broth in the presence of anti- rpoD PPNA2332 and scrambled PPNA2332 at different concentrations with MRSA/VISA Mu50 cells, respectively. The anti- rpoD PPNA2332 was added once at the start of the growth period. After 18 h of growth, total RNA and protein was isolated from treated and untreated cultures. (A) Cellular levels of σ 70 RNA were determined by RT-PCR. The reduction in the amount of RT-PCR product corresponding to gene rpoD in cells treated with anti- rpoD PPNA2332 at different concentrations was determined as the product specific for rpoD RNA relative to products for 16S rRNA . (B) Expressions of σ 70 protein in different treatment were analyzed by western blotting and were quantitated densitometrically.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques: Isolation, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: The symbols represent time points of cell harvest (2, 6, and 24 h) and are displayed as the mean number of colony-forming units (CFUs) per milliliter. Error bar represents SD. The 6 h counts for 1 µM PPNA2332 were 0 CFUs/mL. For all observations in co-cultures (GEP+MRSA/VISA Mu50) treated, with the limit of detection indicated as 10 CFUs/mL. Scr PPNA2332, Scrambled sequence control PPNA2332; GEP, gastric mucosa originated epithelial cell.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques: Sequencing, Control

Journal: PLoS ONE

Article Title: Targeting RNA Polymerase Primary σ 70 as a Therapeutic Strategy against Methicillin-Resistant Staphylococcus aureus by Antisense Peptide Nucleic Acid

doi: 10.1371/journal.pone.0029886

Figure Lengend Snippet: The images show epithelial cell cultures grown in DMEM/HIGH Glucose, 10% FCS. The column at left shows epithelial cell cultures without added MRSA/VISA Mu50; the right column shows epithelial cell cultures that were inoculated with MRSA/VISA Mu50. The top panels of each column show cultures not treated with PPNA, and the rows below show cultures treated with increasing amounts of anti- rpoD PPNA2332 (1, 5, and 10 µM). Magnification, ×100.

Article Snippet: MRSA/VISA Mu50 (ATCC700699) was purchased from MicroBiologics (Minnesota, USA).

Techniques:

Journal: Toxins

Article Title: Prophage-Encoded Staphylococcal Enterotoxin A: Regulation of Production in Staphylococcus aureus Strains Representing Different Sea Regions

doi: 10.3390/toxins7124889

Figure Lengend Snippet: S. aureus strain Mu50 grown in BHI under control and MMC induced conditions. Average values including standard deviations ( ┬ , ┴ ) of three technical replicates are presented. In MMC induced culture the point of induction at 4 h is indicated with an arrow. Time is represented in the X -axis in hours (h). ( A ) Ratio of RF and sea copy levels ( Y -axis) as calculated based on Cq values and according to the equation ratio = (E target ) ΔCq target(control-sample) . Bars represent RF copy levels; dark grey bars designate RF levels for the control culture while light grey bars for the MMC induced culture. The sea 1 gene copy levels are represented by symbols. The levels for the control culture are designated the symbol (▲), while the levels for the MMC induced culture are designated the symbol (×); ( B ) Ratio of total sea 1 transcript represented in dark grey bars for the control culture and light grey bars for the MMC induced. The cro and long sea 1 transcripts were not detected in either the control or MMC induced cultures.

Article Snippet: Four food isolates of S. aureus , Sa17, Sa21, Sa48, and Sa51 (donated from SP Food and Bioscience, the former Swedish Institute for Food and Biotechnology, SIK, Göteborg, Sweden), and a fully sequenced strain Mu50 (LGC Promochem, London, UK), were studied.

Techniques: Control

Journal: Toxins

Article Title: Prophage-Encoded Staphylococcal Enterotoxin A: Regulation of Production in Staphylococcus aureus Strains Representing Different Sea Regions

doi: 10.3390/toxins7124889

Figure Lengend Snippet: S. aureus strains used for analysis of the 3.6 kb gene region including the sea gene.

Article Snippet: Four food isolates of S. aureus , Sa17, Sa21, Sa48, and Sa51 (donated from SP Food and Bioscience, the former Swedish Institute for Food and Biotechnology, SIK, Göteborg, Sweden), and a fully sequenced strain Mu50 (LGC Promochem, London, UK), were studied.

Techniques: