Journal: Journal of Innate Immunity

Article Title: A Common Genetic Variation in Langerin (CD207) Compromises Cellular Uptake of Staphylococcus aureus

doi: 10.1159/000500547

Figure Lengend Snippet: Strains used in this study [ 12 , 32 , 33 , 34 , 35 ]

Article Snippet: S. pyogenes MGAS315 [ 12 ] (Table ) was grown overnight at 37°C without agitation in 5 mL THB (Becton Dickinson) supplemented with 1% yeast extract (Oxoid). table ft1 table-wrap mode="anchored" t5 Table 1 caption a7 Strain Source S. aureus USA300 (NRS384, CC8) NARSA strain collection S. aureus USA300 Δ tarM (WTA α-GlcNAc deficient) [ 32 ] S. aureus Newman (CC8) ATCC, cat. No. 13420 S. aureus Newman Δ spa Δ sbi pCM29 (sGFP expressing) [ 12 ] S. aureus 82086 (CC398) [ 32 ] S. aureus PS66 (CC30) Udo Bläsi, Vienna S. aureus MW2 (CC1) [ 33 ] S. aureus Wood46 (CC97) ATCC, cat. No. 10832 S. aureus Mu50 (CC5) [ 33 ] S. aureus P68 (CC25) Udo Bläsi, Vienna S. aureus NRS184 (CC22) NARSA strain collection S. aureus JH1 (CC5) [ 34 ] S. pyogenes MGAS315 [ 35 ] Open in a separate window Strains used in this study [ 12 , 32 , 33 , 34 , 35 ] Production of Recombinant Langerin Extracellular Domains The extracellular domains of truncated human langerin (residues 148–328) and SNP variants thereof were recombinantly expressed from codon-optimized constructs containing a C-terminal TEV cleavage site followed by a Strep-tag II cloned into pUC19 expression vectors, expressed in E. coli BL21 (DE3; Thermo Fisher), purified and labeled as described previously [ 3 ].

Techniques: Expressing

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Minimal inhibitory concentration of the antibiotics methicillin (Meth), ceftazidime (Ceft), ciprofloxacin (Cip), vancomycin (Van), doxycycline (Doxy), amikacin (Amik), erythromycin (Erythro) and the compounds diethyldithiocarbamate (DDC) and Cu 2+ towards planktonic S. aureus , MRSA, S. epidermidis , E. coli and P. aeruginosa.

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques: Concentration Assay

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Effect of diethyldithiocarbamate (DDC) and Cu 2+ concentrations (in μg/ml) on the viability of (A) MRSA Mu50, (B) MRSA 2, (C) Staphylococcus epidermidis ATCC 35984 and (D) S. epidermidis ATCC 14990 biofilms compared to monotherapy with Cu 2+ ( n = 3; * p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques:

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Synergistic effects of diethyldithiocarbamate in combination with Cu 2+ against S. aureus, MRSA and S. epidermidis biofilms.

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques:

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Minimal concentration to kill over 80% biofilm and synergistic effects of antibiotics, diethyldithiocarbamate and Cu 2+ (DDC-Cu 2+ ) and the combination against MRSA Mu50 ( n = 3).

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques: Concentration Assay

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Comparison of stained MRSA Mu50 and S. epidermidis ATCC 35984 biofilms with LIVE/DEAD BacLight staining after treatment with 8 μg/ml diethyldithiocarbamate and 32 μg/ml Cu 2+ (DDC-Cu 2+ ). Confocal microscopy images results: green = viable bacteria; red = dead bacteria. (A) Untreated S. epidermidis ATCC 35984 biofilm at 20×. S. epidermidis ATCC 35984 biofilm after treatment with DDC-Cu 2+ at (B) 20× and (D) 100×. (C) Quantification of images as green/red ratio of untreated control (black) and treatment with DDC-Cu 2+ (grey) of MRSA and S. epidermidis ATCC 35984 biofilms ( n = 3–8; *** p < 0.001).

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques: Staining, Confocal Microscopy

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Effect of 8 μg/ml diethyldithiocarbamate (DDC; orange), 32 μg/ml Cu 2+ (blue) and combined DDC-Cu 2+ (grey) on (A) the cell index of MRSA Mu50 and (C) S. epidermidis ATCC 35984 over 48 h compared to the untreated control (black). Comparison of the mean cell index between 12 and 48 h for each treatment of (B) MRSA Mu50 and (D) S. epidermidis ATCC 35984 ( n > 3; *** p < 0.001).

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques:

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Monitoring of MRSA Mu50 biofilm formation over 24 h when left untreated or treated with a combination of 8 μg/ml diethyldithiocarbamate and 32 μg/ml Cu 2+ combination (DDC-Cu 2+ ) using the Bioflux system. Scale bar represents 50 μm.

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques:

Journal: Frontiers in Microbiology

Article Title: The combination of diethyldithiocarbamate and copper ions is active against Staphylococcus aureus and Staphylococcus epidermidis biofilms in vitro and in vivo

doi: 10.3389/fmicb.2022.999893

Figure Lengend Snippet: Effect of diethyldithiocarbamate [DDC; orange; 8 μg/ml (A) , 6.4 mg/kg (B–D) ], Cu 2+ [blue; 32 μg/ml (A) , 25.6 mg/kg (B–D) ] and DDC-Cu 2+ (grey) on (A) fibroblast viability ( n = 3), on (B) probability of Galleria mellonella survival (30/group; n = 120), on the probability of survival of Galleria mellonella infected with (C) MRSA Mu50 (30/group; n = 120), and (D) infected with S. epidermidis ATCC 35984 (30/group; n = 120; NS = not significant; * p < 0.05; *** p < 0.001).

Article Snippet: Bioflux plates were primed with 350 μl half-strength tryptone soy broth (TSB, BD, Sparks, MD, United States) and inoculated with 70 μl of a bacterial overnight culture (either MRSA Mu50 or S. epidermidis ATCC 35984) adjusted to OD 600 of 0.2.

Techniques: Infection