Journal: bioRxiv

Article Title: Treatment of wound infections in a mouse model using Zn 2+ -releasing phage bound to gold nanorods

doi: 10.1101/2022.01.05.475129

Figure Lengend Snippet: Treatment of a P. aeruginosa -infected wound by phage-conjugated gold nanorods decorated with Zn 2+ . Treatment is initiated 0-2 days after wound inoculation. The phage-based reagent is applied to the wound and allowed to bind for 30 min. The wound is irradiated with near-infrared light for 15 min to trigger photothermal ablation of P. aeruginosa and release of Zn 2+ ; irradiation may be repeated the next day. Wound healing is observed over the next ∼10 days.

Article Snippet: Reagents were obtained from the following sources: gold(III) chloride trihydrate (HAuCl 3 , 99.9%, Sigma), sodium borohydride (NaBH 4 , 98%, Fisher Scientific), trisodium citrate dihydrate (99.9%; Sigma), P. aeruginosa (Schroeter) Migula (ATCC 25102; PAK strain), P. aeruginosa PAK-pmrB (gift of Prof. Jian Li, Monash University), V. cholerae 0395 (gift of Prof. Michael J. Mahan, UCSB), M13KE phage (New England Biolabs), M13-NotI-Kan construct , sodium chloride (NaCl, 99%, Fisher BioReagents), tryptone (99%, Fisher BioReagents), yeast extract (99%, Fisher BioReagents), E. coli ER2738 (New England Biolabs), N -(3- (dimethylamino)propyl)- N′ -ethylcarbodiimide hydrochloride (EDC, 99%, Sigma), N- hydroxysuccinimide (NHS, 98%, Sigma), N -succinimidyl- S -acetylthiopropionate (SATP) (Thermo Fisher Scientific), 5-bromosalicylic acid (5-BAA) (>98.0%; TCI), isopropyl β-D-1- thiogalactopyranoside (IPTG) (99%; Fisher Scientific), thiol-PEG-acid (HOOC-PEG-SH; PEG average M n 5,000; Sigma), poly(ethylene glycol) (PEG-8000, Sigma), dialysis kit (MWCO 3500 Da, Spectrum Laboratories), tetracycline (Sigma), kanamycin sulfate (Sigma), Top 10F′ cyan cells (Thermo Fisher), Mix and Go competent cells (Zymo Research), QIAprep Spin Miniprep Kit (Qiagen), QIAquick Gel Extraction Kit (Qiagen), and KpnI-HF/NotI-HF restriction enzyme and T4 DNA ligase (New England Biolabs), Pseudomonas aeruginosa antibody (1010/287) (Novus Biologicals, LLC).

Techniques: Infection, Irradiation

Journal: bioRxiv

Article Title: Treatment of wound infections in a mouse model using Zn 2+ -releasing phage bound to gold nanorods

doi: 10.1101/2022.01.05.475129

Figure Lengend Snippet: Antibacterial effect of phanorods and phanorod-Zn in vitro , measured by colony- forming units. P. aeruginosa cells in suspension were exposed to phanorods and phanorod-Zn without irradiation. The presence of these reagents did not affect bacterial survival (a) (control = no phanorods or phanorod-Zn added). With NIR laser irradiation, cell survival was reduced in the presence of phanorods, and phanorod-Zn showed a significantly greater reduction in cell survival (b). If repeated washing was applied to remove released Zn 2+ (c), the colony counts (with NIR laser irradiation) were similar between phanorods and phanorod-Zn exposure. Colony counts of a treated P. aeruginosa biofilm (d) show trends similar to P. aeruginosa in suspension (b). Error bars are standard deviation (n = 5): ** p < 0.01, *** p < 0.001, NS, not significant ( p > 0.05, two-sided t test).

Article Snippet: Reagents were obtained from the following sources: gold(III) chloride trihydrate (HAuCl 3 , 99.9%, Sigma), sodium borohydride (NaBH 4 , 98%, Fisher Scientific), trisodium citrate dihydrate (99.9%; Sigma), P. aeruginosa (Schroeter) Migula (ATCC 25102; PAK strain), P. aeruginosa PAK-pmrB (gift of Prof. Jian Li, Monash University), V. cholerae 0395 (gift of Prof. Michael J. Mahan, UCSB), M13KE phage (New England Biolabs), M13-NotI-Kan construct , sodium chloride (NaCl, 99%, Fisher BioReagents), tryptone (99%, Fisher BioReagents), yeast extract (99%, Fisher BioReagents), E. coli ER2738 (New England Biolabs), N -(3- (dimethylamino)propyl)- N′ -ethylcarbodiimide hydrochloride (EDC, 99%, Sigma), N- hydroxysuccinimide (NHS, 98%, Sigma), N -succinimidyl- S -acetylthiopropionate (SATP) (Thermo Fisher Scientific), 5-bromosalicylic acid (5-BAA) (>98.0%; TCI), isopropyl β-D-1- thiogalactopyranoside (IPTG) (99%; Fisher Scientific), thiol-PEG-acid (HOOC-PEG-SH; PEG average M n 5,000; Sigma), poly(ethylene glycol) (PEG-8000, Sigma), dialysis kit (MWCO 3500 Da, Spectrum Laboratories), tetracycline (Sigma), kanamycin sulfate (Sigma), Top 10F′ cyan cells (Thermo Fisher), Mix and Go competent cells (Zymo Research), QIAprep Spin Miniprep Kit (Qiagen), QIAquick Gel Extraction Kit (Qiagen), and KpnI-HF/NotI-HF restriction enzyme and T4 DNA ligase (New England Biolabs), Pseudomonas aeruginosa antibody (1010/287) (Novus Biologicals, LLC).

Techniques: In Vitro, Suspension, Irradiation, Control, Standard Deviation

Journal: bioRxiv

Article Title: Treatment of wound infections in a mouse model using Zn 2+ -releasing phage bound to gold nanorods

doi: 10.1101/2022.01.05.475129

Figure Lengend Snippet: Treatment of P. aeruginosa -infected wounds by phanorod-Zn in vivo . (a) Thermal images of control and phanorod-Zn treatment before and after the NIR laser irradiation. (b) Representative photographs of the infected wound over time for control, Zn 2+ ion, phanorod, and phanorod-Zn treatment (scale bar = 5 mm). (c) Wound area over time for control, phanorod, and phanorod-Zn treatment. Also see Figure S19. (d) Bacterial load assessed by colony-forming units from wound tissue on days 2 and 4. Error bars show standard deviation (n = 5 mice): *** indicates p < 0.001; NS = not significant ( p > 0.05); two-sided t test.

Article Snippet: Reagents were obtained from the following sources: gold(III) chloride trihydrate (HAuCl 3 , 99.9%, Sigma), sodium borohydride (NaBH 4 , 98%, Fisher Scientific), trisodium citrate dihydrate (99.9%; Sigma), P. aeruginosa (Schroeter) Migula (ATCC 25102; PAK strain), P. aeruginosa PAK-pmrB (gift of Prof. Jian Li, Monash University), V. cholerae 0395 (gift of Prof. Michael J. Mahan, UCSB), M13KE phage (New England Biolabs), M13-NotI-Kan construct , sodium chloride (NaCl, 99%, Fisher BioReagents), tryptone (99%, Fisher BioReagents), yeast extract (99%, Fisher BioReagents), E. coli ER2738 (New England Biolabs), N -(3- (dimethylamino)propyl)- N′ -ethylcarbodiimide hydrochloride (EDC, 99%, Sigma), N- hydroxysuccinimide (NHS, 98%, Sigma), N -succinimidyl- S -acetylthiopropionate (SATP) (Thermo Fisher Scientific), 5-bromosalicylic acid (5-BAA) (>98.0%; TCI), isopropyl β-D-1- thiogalactopyranoside (IPTG) (99%; Fisher Scientific), thiol-PEG-acid (HOOC-PEG-SH; PEG average M n 5,000; Sigma), poly(ethylene glycol) (PEG-8000, Sigma), dialysis kit (MWCO 3500 Da, Spectrum Laboratories), tetracycline (Sigma), kanamycin sulfate (Sigma), Top 10F′ cyan cells (Thermo Fisher), Mix and Go competent cells (Zymo Research), QIAprep Spin Miniprep Kit (Qiagen), QIAquick Gel Extraction Kit (Qiagen), and KpnI-HF/NotI-HF restriction enzyme and T4 DNA ligase (New England Biolabs), Pseudomonas aeruginosa antibody (1010/287) (Novus Biologicals, LLC).

Techniques: Infection, In Vivo, Control, Irradiation, Standard Deviation

Journal: bioRxiv

Article Title: Treatment of wound infections in a mouse model using Zn 2+ -releasing phage bound to gold nanorods

doi: 10.1101/2022.01.05.475129

Figure Lengend Snippet: Treatment of wounds infected by polymyxin-resistant P. aeruginosa . (a) Wounds assessed by size over time, with polymyxin treatment initiated on day 0, or phanorod-Zn treatment initiated on day 0 (L0+L1) or day 2 (L2+L3). (b) Quantitative assessment of bacterial load (cfu) for polymyxin-resistant P. aeruginosa wound infection, measured on days 2, 4 and 6. For delayed treatment (L2+L3), samples on day 2 were taken without treatment. Error bars show standard deviation (n = 5 mice): *** indicates p < 0.001; NS = not significant ( p > 0.05); two- sided t test.

Article Snippet: Reagents were obtained from the following sources: gold(III) chloride trihydrate (HAuCl 3 , 99.9%, Sigma), sodium borohydride (NaBH 4 , 98%, Fisher Scientific), trisodium citrate dihydrate (99.9%; Sigma), P. aeruginosa (Schroeter) Migula (ATCC 25102; PAK strain), P. aeruginosa PAK-pmrB (gift of Prof. Jian Li, Monash University), V. cholerae 0395 (gift of Prof. Michael J. Mahan, UCSB), M13KE phage (New England Biolabs), M13-NotI-Kan construct , sodium chloride (NaCl, 99%, Fisher BioReagents), tryptone (99%, Fisher BioReagents), yeast extract (99%, Fisher BioReagents), E. coli ER2738 (New England Biolabs), N -(3- (dimethylamino)propyl)- N′ -ethylcarbodiimide hydrochloride (EDC, 99%, Sigma), N- hydroxysuccinimide (NHS, 98%, Sigma), N -succinimidyl- S -acetylthiopropionate (SATP) (Thermo Fisher Scientific), 5-bromosalicylic acid (5-BAA) (>98.0%; TCI), isopropyl β-D-1- thiogalactopyranoside (IPTG) (99%; Fisher Scientific), thiol-PEG-acid (HOOC-PEG-SH; PEG average M n 5,000; Sigma), poly(ethylene glycol) (PEG-8000, Sigma), dialysis kit (MWCO 3500 Da, Spectrum Laboratories), tetracycline (Sigma), kanamycin sulfate (Sigma), Top 10F′ cyan cells (Thermo Fisher), Mix and Go competent cells (Zymo Research), QIAprep Spin Miniprep Kit (Qiagen), QIAquick Gel Extraction Kit (Qiagen), and KpnI-HF/NotI-HF restriction enzyme and T4 DNA ligase (New England Biolabs), Pseudomonas aeruginosa antibody (1010/287) (Novus Biologicals, LLC).

Techniques: Infection, Standard Deviation

Journal:

Article Title: Chemokines and activated macrophages in HIV gp120-induced neuronal apoptosis

doi:

Figure Lengend Snippet: Protection of rat cerebrocortical neurons from gp120-induced apoptosis by the β-chemokines RANTES and MIP-1β. (A) Neuroprotection by recombinant rat RANTES (20 nM). (B) Neuroprotection by recombinant human MIP-1β (20 nM). Rat cerebrocortical cultures were incubated for 24 hr with or without 200 pM recombinant gp120 and in the presence or absence of each chemokine. After fixation and permeabilization, neurons were identified by immunostaining for MAP-2 or NeuN, and apoptotic cells were assessed by propidium iodide staining. ∗, P < 0.01 compared with value for gp120.

Article Snippet: Recombinant human MIP-1β, SDF-1α, SDF-1β, and recombinant rat RANTES were purchased from R&D Systems and Endogen (Cambridge, MA), respectively.

Techniques: Recombinant, Incubation, Immunostaining, Staining