CC5 isolates (n = 43) characterized by <i>spa</i>-typing and comprehensive MA subgroup analysis using three different bioinformatic modes (principal component analyses, splits graph and cluster dendrogram).

<p>CC5 isolates (n = 43) characterized by <i>spa</i>-typing and comprehensive MA subgroup analysis using three different bioinformatic modes (principal component analyses, splits graph and cluster dendrogram).</p

Risk factors of MRSA and matched MSSA control group isolates.

#<p>statistical analysis was not performed for clinical criteria applied for selection of matched MSSA cases, ns =  not significant.</p

Principal component cluster analysis (PCA) of 41 MRSA (R1–R41) and two MSSA (S42, S43) of CC5.

<p>(A) Clustering of the 43 CC5 isolates by PCA as well as (B) subclustering of 30 MRSA CC5 cluster I isolates using a higher resolution PCA plot for in-depth identification of additional subgroups (Ia–Id).</p

Diversity analysis of all MSSA (S1–S46) and MRSA (R1–R46) isolates by splits graph.

<p>(A) Splits graph constructed based on cost distance matrix produced by Ridom StaphType and (B) on default settings of the IdentiBAC microarray hybridization profiles of 334 genes and alleles. Clonal complexes (CC) as well as the most abundant <i>spa</i>-types t003 (circles) and t012 (quadrates) were highlighted.</p

Subclassification analysis of 41 MRSA (R1–R41) and two MSSA (S42, S43) of CC5.

<p>(A) Splits graph based on cost distance matrix computed by Ridom StaphType software. (B) Splits graph based on MA hybridization profiles. Characteristic gene profiles for isolate cluster assignment were arbitrarily stated into group A-E. The most common MRSA <i>spa</i>-types t003 (circles), t504 (quadrates) and t010 (hexagons) were highlighed.</p

Glucose and acetate contents in the supernatants of growing <i>S</i>. <i>aureus</i> COL cultures.

<p>A) Consumption of glucose in the presence (●) and absence (■) of kendomycin. B) Accumulation/consumption of acetate in the presence (●) and absence (■) of kendomycin. Arrows indicate the addition of kendomycin. The data presented are the mean ± SD of three independent experiments.</p

Effect of kendomycin on the cell division in <i>S</i>. <i>aureus</i> COL.

<p>Electron microscopic images of healthy control cells (A, B) and kendomycin treated cells (C-F). (A) Dividing control cells at the mid of septum formation. (B) Dividing cells with a completed septum. (C-F)Kendomycin treated cells exhibiting irregular septum morphologies. Bar = 200 nm.</p

Effect of kendomycin on growth of <i>S</i>. <i>aureus</i> strain COL.

<p>Bacterial cultures were grown aerobically in BHI medium, and the optical densities at 600 nm of the cultures were measured at the time points indicated. Kendomycin addition is displayed by vertical arrows. Negative controls (■) were challenged with the solvent methanol at concentrations equivalent to the highest concentration added via the kendomycin supplementation. A) Effect of different kendomycin concentrations on growth of strain COL: 0.5 x MIC (●), 1x MIC (▲), and 3 x MIC (▼). The data presented are the mean ± SD of three independent experiments. B) Impact of kendomycin on the long-term growth characteristic of the <i>S</i>. <i>aureus</i> COL cell culture. BHI cultures of strain COL were grown for up to 24 h in absence (■) or presence of 3 x MIC kendomycin (●). The data presented are the mean ± SD of three independent experiments. C) Effect of kendomycin supplementation on the viability of the COL cell culture. Mid-exponential growth phase cells were either left untreated or challenged with 1.5 x MIC of kendomycin (●), and grown as outlined before. The number of viable bacteria were determined at the time points indicated by plating out serial dilutions of the cultures on agar plates. The data presented are the mean ± SD of two independent experiments.</p

Effects of Kendomycin on the synthesis of macromolecules.

<p>The radioactive labeled precursors ³H-glucosamine (A), ¹⁴C-isoleucine (B), ¹⁴C-thymidin (C), and ³H-uridin (D) were added to an exponential growth phase culture of <i>S</i>. <i>aureus</i> strain COL in absence (■) and presence (●) of kendomycin (5 x MIC). Incorporation of labeled substances was determined as outlined in materials and methods. The data presented are the mean ± SD of two independent experiments.</p

Effect of kendomycin on the transcription of selected genes in <i>S</i>. <i>aureus</i> COL.

<p><i>S</i>. <i>aureus</i> COL cells were grown in BHI medium to mid-exponential growth phase, and subsequently supplemented with kendomycin (1.5 x MIC, white bars) or left untreated (black bars). One hour after drug addition, cells were harvested and used for total RNA isolation. A) Transcription of the heat shock protein encoding genes <i>clpC</i>, <i>clpB</i> and <i>clpP</i>. B) Transcription of the capsule operon genes <i>cap5A</i> and <i>cap8C</i>, the UDP-N-acetylglucosamine1-caboxyphenyltransferase encoding gene <i>murAA</i>, and the septum formation factors encoding genes <i>ftsA</i> and <i>ftsZ</i>. C) Transcription of the programmed cell death factor encoding genes <i>cidA</i> and <i>lrgA</i>. D) Transcription of the Sigma^B activity marker gene <i>asp23</i>. E) Transcription of the H₂O₂ inactivating factor encoding genes <i>ahpC</i> and <i>katA</i>. mRNA levels are expressed relative to gyrase B (in numbers of copies per copy of <i>gyrB</i>). The data presented are the mean ± SD of four independent experiments.</p