Homologies of gene <i>rpoD</i> among <i>Staphylococcus</i> species.

<p>Homologies of gene <i>rpoD</i> among <i>Staphylococcus</i> species.</p

Binding parameters predicted by software Oligo walk 5.0 and PNALIGHT.

a<p>Numbering from the first base of the gene <i>rpoD</i>;</p>b<p>ΔG means free energy; index for each parameter: GC%≤60%, overall ΔG<−10 kcal/mol, Duplex ΔG<−25 kcal/mol, oligo-self ΔG≥−1.1 kcal/mol, oligo-oligo ΔG≥−8 kcal/mol, Tm>50°C;</p>c<p>“—” means not determined;</p>d<p>Calculated melting temperature in [°C] of matching PNA/DNA hybrids with no dangling ends.</p

MIC of anti-<i>rpoD</i> peptide-PNAs for quality control and clinical strains of <i>Staphyloccus aureus</i> in M-H broth culture.

a<p><i>rpoD</i>, RNA polymerase sigma 70; PNA, peptide nucleic acid; PPNA means peptide conjugated PNA; The PNAs are written from their N to their C terminus, and the N terminus corresponds to the 5′ end of a conventional oligonucleotide; “K” indicates lysine, F indicates phenylalanine, “X” indicates 6-aminohexanoic acid, “B” indicates β-alanine, and eg¹ indicates glycine; Scr means PNA with a scrambled base sequence (as control);</p>b<p>Minimal inhibitory concentrations (MIC) were the lowest PNA concentrations that prevented bacterial growth by visual inspection after overnight (24 h) growth from an inoculum of 10⁵ CFU/mL. “VISA” is abbreviation for vancomycin-intermediate resistance <i>Staphyloccus aureus</i>; “MRSA” is abbreviation for methicillin-resistant <i>Staphyloccus aureus</i>; XIJING means clinical MRSA isolate from patients in Fourth Military Medical University affiliated XIJING hospital.</p

Protective effect of anti-<i>rpoD</i> PPNA2332 on epithelial cell culture infected with noninvasive single MRSA/VISA Mu50 infection.

<p>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.</p

Bacteriocidal effects of anti-<i>rpoD</i> PPNA2332 on the viable cells of MRSA/VISA Mu50 in pure culture.

<p>Anti-<i>rpoD</i> PPNA2332 was added to cell cultures containing 1.0×10⁵ 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)₃K (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.</p

Light micrographs of epithelial cell cultures.

<p>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-<i>rpoD</i> PPNA2332 (1, 5, and 10 µM). Magnification, ×100.</p

Effects of anti-<i>rpoD</i> PPNA2332 on the growth of MRSA/VISA Mu50 in pure culture.

<p>Anti-<i>rpoD</i> PPNA2332 was added to cell cultures containing 1.0×10⁵ 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)₃K (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.</p

DMB activates the GLP-1 receptor in cortical neurons.

<p>(A) Western blot showing GLP-1R expression in neurons and β-cells. (B) Bar graph reflecting the GLP-1R expression in each group. Data are presented as percentage of β-cells. n = 5 (C) cAMP stimulation by DMB (1 μM) in neurons and detected 0–30 min after incubation. Data are expressed as percentage of 0 min. n = 6.</p

Structure of DMB, molecular formula C₁₃H₁₅Cl₂N₃O₂S.

<p>Structure of DMB, molecular formula C₁₃H₁₅Cl₂N₃O₂S.</p

DMB treatment decreased neuronal apoptosis in cerebral ischemia.

<p>(A) TUNEL assay 24 h post MCAO or sham operation. Photographs were taken from ipsilateral ischemic penumbra area. Scale bar = 25 μm. (B) Bar graph reflects the number of TUNEL-positive neurons in each group. Six different fields were counted. (C) Western blot assay of Bax and Bcl-2 expression in ipsilateral ischemic penumbra area 24 h post MCAO or sham operation. (D) Bar graph reflects Bax/Bcl-2 ratio in each group. n = 6. ** <i>p</i> < 0.01.</p