Journal: Frontiers in Microbiology

Article Title: Genome-wide transcription response of Staphylococcus epidermidis to heat shock and medically relevant glucose levels

doi: 10.3389/fmicb.2024.1408796

Figure Lengend Snippet: Environmental perturbation of S. epidermidis . Log-phase cultures were exposed to (A) a 10-min increase in temperature from 37 to 45°C or (B) a range of 20-min glucose spikes (concentrations as noted) and a 10 mM spike followed by a step down to 2 mM.

Article Snippet: As a second example, Azitra, Inc. indicates they are engineering S. epidermidis strains to deliver therapeutic proteins to treat skin diseases including Netherton Syndrome and to improve skin appearance ( ).

Techniques:

Journal: Frontiers in Microbiology

Article Title: Genome-wide transcription response of Staphylococcus epidermidis to heat shock and medically relevant glucose levels

doi: 10.3389/fmicb.2024.1408796

Figure Lengend Snippet: A sudden temperature increase causes transcript levels to change up to ~ 71-fold. (A) Volcano plot showing the differentially expressed genes (DEGs) for the heat-shock experimental condition with |log2 FC| ≥ 1.5 and adjusted p ≤ 0.05 as the threshold. The red dots represent 235 significantly upregulated genes, and the blue dots represent 131 significantly downregulated genes. (B) Summary of the significantly upregulated and downregulated genes during the heat-shock response in S. epidermidis assigned to functional groups according to GO and KEGG pathways (in %).

Article Snippet: As a second example, Azitra, Inc. indicates they are engineering S. epidermidis strains to deliver therapeutic proteins to treat skin diseases including Netherton Syndrome and to improve skin appearance ( ).

Techniques: Functional Assay

Journal: Frontiers in Microbiology

Article Title: Genome-wide transcription response of Staphylococcus epidermidis to heat shock and medically relevant glucose levels

doi: 10.3389/fmicb.2024.1408796

Figure Lengend Snippet: Eighty-five S. epidermidis genes change expression levels in response to glucose. (A) Circular transcriptome map showing normalized gene expression levels in the S. epidermidis genome in response to glucose. Log2 fold change relative to control for cells exposed to 2 mM (G2), 5 mM (G5), 10 mM (G10), 17 mM (G17), or 50 mM (G50) glucose spikes. Each bar denotes a single gene; red bars represent significantly upregulated genes and blue bars represent significantly downregulated genes. Roman numerals i ( sdaAB , rbsU ), ii ( pflB ), iii ( glpR-pfkB operon), iv (F1613_RS07845 (homoserine dehydrogenase), and v (members of the lac operon) correspond to select groups of genes that are downregulated across all five glucose spike conditions. (B) Glucose concentration-response curves for a representative subset of genes that have potentially interesting glucose-responsive switch properties.

Article Snippet: As a second example, Azitra, Inc. indicates they are engineering S. epidermidis strains to deliver therapeutic proteins to treat skin diseases including Netherton Syndrome and to improve skin appearance ( ).

Techniques: Expressing, Gene Expression, Control, Concentration Assay

Journal: Frontiers in Microbiology

Article Title: Genome-wide transcription response of Staphylococcus epidermidis to heat shock and medically relevant glucose levels

doi: 10.3389/fmicb.2024.1408796

Figure Lengend Snippet: The genome-wide transcription response of S. epidermidis to perturbations. Circular transcriptome map showing normalized gene expression levels in the S. epidermidis genome. Log2-fold change relative to control for cells exposed to Heat Shock (HS), a 17 mM glucose spike (G17), or Step Down (SD) experimental conditions. Each bar denotes a single gene. The red bars represent significantly upregulated genes, and the blue bars represent significantly downregulated genes.

Article Snippet: As a second example, Azitra, Inc. indicates they are engineering S. epidermidis strains to deliver therapeutic proteins to treat skin diseases including Netherton Syndrome and to improve skin appearance ( ).

Techniques: Genome Wide, Gene Expression, Control

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Bacteriophage Lysin CF-301, a Potent Antistaphylococcal Biofilm Agent

doi: 10.1128/AAC.02666-16

Figure Lengend Snippet: Formation of mixed-species biofilms and analysis of CF-301 susceptibility. Biofilms containing S. aureus (strain BAA-42) and/or S. epidermidis (strain NRS 7) were grown either on 24-well polystyrene plates for 3 days or on catheters and surgical mesh for 6 days before treatment with a 10-fold dilution series of CF-301. (A) Quantitation of S. aureus and S. epidermidis CFU in mixed biofilms prior to CF-301 treatment. Experiments were performed in triplicate, and CFU represent the mean ± standard deviation. (B) Crystal violet staining of single- and mixed-species biofilms on 24-well plates treated for 24 h with CF-301 (relative to untreated control). (C) Catheter biofilms treated for 4 h. (D) Catheter biofilms treated for 24 h. (E) Surgical mesh biofilms treated for 24 h.

Article Snippet: The strains used in this study include S. aureus ATCC BAA-42 (American Type Culture Collection [ATCC], Manassas, VA), S. aureus Newman ( 65 ), S. aureus MW2 (NRS 123; BEI Resources), S. aureus CFS-1246 (Newman/GFP plasmid pCN57 [ 66 ] derived from NRS 623 [BEI Resources]), S. aureus VRS3a (NR-46412; BEI Resources), and S. epidermidis NRS 7 (BEI Resources).

Techniques: Quantitation Assay, Standard Deviation, Staining

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: PSMs in culture filtrates of S. aureus and S. epidermidis induce neutrophil degranulation and NE release. Neutrophils (polymorphonuclear leukocytes=PMNs) were stimulated with (A) culture filtrates of S. aureus (0,75%), S. epidermidis (3%) or the indicated isogenic PSM or agr mutant (B) with fMLF, PSMα2, PSMα3 or δ-toxin or (C) with S. epidermidis PSMα, PSMδ or PSMε, and degranulation (β-glucuronidase release) was analyzed. (D) Neutrophils were stimulated with the indicated S. aureus or (E) S. epidermidis PSMs and elastase activity was analyzed. Data represents mean and SEM of at least three independent experiments. ns, not significant, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 significant difference versus the indicated wild-type strains as calculated by paired two-tailed Student’s t tests (A) or one-way ANOVA with Dunnett’s multiple comparisons test (B–E) .

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques: Mutagenesis, Activity Assay, Two Tailed Test

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: Neutrophils inactivate PSM-mediated FPR2 activity of culture filtrates by NSPs. Culture filtrates of S. aureus USA300 (A) or S. epidermidis RN1457 (B) were incubated for six hours with increasing amounts of neutrophils. FPR2 transfected HL60 were stimulated with these treated culture filtrates and FPR2 activity was analyzed. PSMα3 (C) or PSMε (D) were incubated with neutrophils. FPR2 transfected HL60 were stimulated with these supernatants and PSM activity was analyzed. Addition of PSMα3 (E) or PSMε (F) to culture filtrates of a S. aureus PSM mutant induces FPR2 activation, an effect that can be dose-dependently inactivated by increasing amounts of neutrophils. Preincubation of neutrophils with the NSP inhibitor PMSF prevents inactivation of FPR2 activity (G) or CD11b upregulation by culture filtrates (H) . Data represents mean and SEM of at least three independent experiments. ns, not significant, *P < 0.05; ***P < 0.001; ****P < 0.0001 significant difference versus the indicated untreated control (A–F, H) or the neutrophil-treated control (G) as calculated by one-way ANOVA with Dunnett’s multiple comparisons test (A–H) .

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques: Activity Assay, Incubation, Transfection, Mutagenesis, Activation Assay, Control

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: S. aureus -derived NSP inhibitors prevent degradation of PSMα3. Western blot of PSMα3 after incubation with NE alone (0.15 U) or with 2 µg/ml Eap (A) , EapH1 (4 µg/ml) (B) or EapH2 (4 µg/ml) (C) . NE activity of neutrophils +/-preincubation with Eap, EapH1 or EapH2 after stimulation with PSMα3 (D) , PSMε (E) . NE activity of neutrophils incubated with S. epidermidis culture filtrates (3%) +/- recombinant Eap, EapH1 or EapH2 (each 5µg/ml). Data represent one experiment out of three (A–C) and mean +/-SEM of three independent experiments (A–D) . *P < 0.05; ***P < 0.001; ****P < 0.0001 significant difference versus NE-treated PSMα3 (A–C) , the PSMα3- (D) or PSMε-treated control (E) as well as the culture filtrate treated control (F) as calculated by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques: Derivative Assay, Western Blot, Incubation, Activity Assay, Recombinant, Control

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: NSPs enhance degradation of PSMs by neutrophils in culture filtrates of USA300. No pre-incubation or prior incubation of culture filtrates of wild-type USA300 with neutrophils for 24 h leads to disappearance of original peaks representative for α and β-PSMs in the HPLC chromatogram, but also to the appearance of new peaks. Treatment of culture filtrates of USA300 ΔeapΔH1ΔH2::ermB with neutrophils only leads to disappearance of nearly all peaks representative for PSMs (A) . Heat map of HPLC/MS Analysis of neutrophil-treated or untreated culture filtrates show that incubation of USA300 indeed leads to disappearance of a few PSM fragments, found in untreated culture filtrates, but also to the appearance of new fragments. In neutrophil-treated culture filtrates of the USA300 ΔeapΔH1ΔH2::ermB mutant most truncated fragments are absent. Heat map represents data normalized towards USA300 (above the line) or towards USA300 plus neutrophils (below the line) (B) . C-terminally truncated PSM fragments with highest abundance in neutrophil-treated culture filtrates of USA300 (C) or PSM fragments with highest abundance in untreated culture filtrates of USA300 (D) . Data represent mean and SEM of three different culture filtrates of USA300 or USA300 ΔeapΔH1ΔH2::ermB , each incubated with or without neutrophils of two different donors. ns, not significant, *P < 0.05; ****P < 0.0001 significant difference versus USA300+neutrophils (C) or USA300 (D) as calculated by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques: Incubation, Mutagenesis

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: The combined deletion of the three NSP inhibitors enhances inactivation of PSMs by neutrophils in culture filtrates of USA300 and lead to enhanced killing of S. aureus. Incubation of neutrophils with bacterial culture filtrates of USA300 and isogenic triple-NSP inhibitor mutant induces release of NE. Inactivation of all three NSP inhibitors strongly enhances NE activity, whereas single knock outs do not (A) . Incubation of neutrophils for 24 h with USA300 or USA300 ΔeapΔH1ΔH2::ermB leads to enhanced inactivation of FPR2 activity (B, C) and cytotoxicity (D, E) of the NSP inhibitor mutant compared to the wild type. The deletion of all three NSP inhibitors but not of a single NSP inhibitor impairs survival of S. aureus in the presence of neutrophils (F) . Killing of S. aureus USA300 compared to isogenic triple-NSP inhibitor mutant and S. epidermidis (G) . Data represent means ± SD of at least three independent experiments. ns, not significant, *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001 significant difference versus USA300 (A, G, H) or untreated controls (B–E) as calculated by one-way ANOVA with Dunnett’s multiple comparisons test (A–G) or unpaired Students t-test (H) .

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques: Incubation, Mutagenesis, Activity Assay

Journal: Frontiers in Immunology

Article Title: Staphylococcus aureus Depends on Eap Proteins for Preventing Degradation of Its Phenol-Soluble Modulin Toxins by Neutrophil Serine Proteases

doi: 10.3389/fimmu.2021.701093

Figure Lengend Snippet: Influence of NSPs on PSMs in S. aureus and S. epidermidis. PSMs of S. aureus and coagulase-negative S. epidermidis activate neutrophils, induce degranulation and release of the NSPs NE, CG and PR3. Active NSPs degrade PSMs of staphylococci. S. aureus , but not the coagulase-negative S. epidermidis secretes three NSP inhibitors, the extracellular adherence proteins (Eap, EapHI, EapHII), which specifically inhibit NSPs. These NSP inhibitors prevent degradation of PSMs and thereby enhance the capability of S. aureus to survive better than S. epidermidis in the presence of neutrophils.

Article Snippet: Formylated S. epidermidis PSMα, PSMε and PSMδ were synthesized by EMC Microcollections, Tuebingen.

Techniques:

Journal: Pathogens

Article Title: The Impact of Intraspecies Variability on Growth Rate and Cellular Metabolic Activity of Bacteria Exposed to Rotating Magnetic Field

doi: 10.3390/pathogens10111427

Figure Lengend Snippet: The effects exerted by different types of magnetic fields on bacteria.

Article Snippet: S. epidermidis , Tessaro et al., 2015 [ ] ELF-EMF , Inhan-Garip et al., 2011 [ ] ELF-EMF Masood et al., 2020 [ ] SMF/O-MF Bajpai et al., 2012 [ ] MI-SMF , Salmen et al., 2018 [ ] HF-EMF .

Techniques: Bacteria

Journal: Cell reports

Article Title: Staphylococcus epidermidis activates keratinocyte cytokine expression and promotes skin inflammation through the production of phenol-soluble modulins

doi: 10.1016/j.celrep.2023.113024

Figure Lengend Snippet:

Article Snippet: S. epidermidis PSMe, fMFIINLVKKVISFIKGLFGNNENE , Lifetein , N/A.

Techniques: Staining, Virus, Recombinant, SYBR Green Assay, cDNA Synthesis, DNA Purification, Enzyme-linked Immunosorbent Assay, Plasmid Preparation, Software