P. aeruginosa strains were pre-grown for 36 h in M9 minimal medium supplemented with 0.5% (v/v) glycerol as carbon source, spun down and resuspended in either M9 glycerol or M9 glycerol containing 500 µM 2,2′-Bipyridine (Fisher Scientific). The bacterial stock solutions were used to inoculate M9 glycerol and M9 glycerol medium containing 500 µM 2,2′-Bipyridine at a starting OD600 of 0.05. Bacterial growth was followed over the course of 96 h in the presence of three different concentrations (0.25 µM, 1 µM and 2 µM) of pyochelin or pyochelin methyl ester or an equivalent volume of methanol. OD600 measurements were taken at 0 h and every 12 h thereafter in an Infinite M PLEX plate reader (Tecan).
2 2 bipyridine
Manufactured by Thermo Fisher Scientific
Sourced in Germany
2,2′-bipyridine is a chemical compound commonly used in analytical and research applications. It functions as a bidentate ligand, capable of coordinating with metal ions to form stable complexes. This property makes it a useful component in various laboratory equipment and procedures, but a detailed description of its intended use would require more specific information about the application context.
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Lab products found in correlation
Ammonium hexafluorophosphate, by Merck Group (2 mentions)
Infinite m plex plate reader, by Tecan (1 mentions)
Synergy ht, by Agilent Technologies (1 mentions)
Calcium nitrate tetrahydrate, by Thermo Fisher Scientific (1 mentions)
Urea, by Thermo Fisher Scientific (1 mentions)
D glucose, by Thermo Fisher Scientific (1 mentions)
Silver nitrate, by Thermo Fisher Scientific (1 mentions)
Boric acid, by Thermo Fisher Scientific (1 mentions)
Nickel 2 nitrate hexahydrate, by Thermo Fisher Scientific (1 mentions)
Strontium nitrate, by Thermo Fisher Scientific (1 mentions)
Magnesium nitrate hexahydrate, by Thermo Fisher Scientific (1 mentions)
Barium nitrate, by Thermo Fisher Scientific (1 mentions)
Cobalt chloride, by Maclin Biochemical Technology (1 mentions)
4 dimethylaminopyridine, by Merck Group (1 mentions)
Dichloromethane, by Avantor (1 mentions)
Triethylamine, by Merck Group (1 mentions)
Tetrahydrofuran, by Avantor (1 mentions)
Ethyl alcohol, by Avantor (1 mentions)
Methyl ether methacrylate, by Merck Group (1 mentions)
Methyl alcohol, by Avantor (1 mentions)
8 protocols using 2 2 bipyridine
1
Pyochelin's effect on P. aeruginosa growth
2
Quantification of Cellular Iron Content
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Following plating, iron loading and washing as above, the PBS was removed and the plates were frozen at −20 °C pre-analysis. Triton X-100 (Sigma-Aldrich, Cat # T8787-50ML, 200 µl at 0.01%) solution was added directly to each well and the cells were harvested with a cell scraper. After micropipetting to encourage lysis, the cell lysates were transferred to microtubes. Cell lysate (10 µl) was removed from each sample for protein quantification using an improved Lowry [29] (link) -type assay (BIO-RAD DC protein assay, Cat # 500-011).
A sensitive and validated colorimetric assay [30] (link) utilising 2,2′-bipyridine (Fisher Scientific, Cat # 11492438) was calibrated and used to quantify total iron content in remaining cell lysate. A standard curve was constructed by adding 10 µl of 10 × 10−3, 6 × 10−3, 4 × 10−3, 1.6 × 10−3, 0.64 × 10−3, and 0.26 × 10−3 M freshly prepared solutions of ammonium iron (II) sulphate hexahydrate (Sigma-Aldrich, Cat # 203505) to ‘control’ cell lysate (no USPIO exposure). Absorbance at 520 nm was measured in a 96-well plate on a Synergy HT (BioTek Instruments Inc., USA) plate reader. Iron concentration was calculated by averaging triplicate samples, expressed as ng/µg protein.
A sensitive and validated colorimetric assay [30] (link) utilising 2,2′-bipyridine (Fisher Scientific, Cat # 11492438) was calibrated and used to quantify total iron content in remaining cell lysate. A standard curve was constructed by adding 10 µl of 10 × 10−3, 6 × 10−3, 4 × 10−3, 1.6 × 10−3, 0.64 × 10−3, and 0.26 × 10−3 M freshly prepared solutions of ammonium iron (II) sulphate hexahydrate (Sigma-Aldrich, Cat # 203505) to ‘control’ cell lysate (no USPIO exposure). Absorbance at 520 nm was measured in a 96-well plate on a Synergy HT (BioTek Instruments Inc., USA) plate reader. Iron concentration was calculated by averaging triplicate samples, expressed as ng/µg protein.
3
Synthesis of Metal Coordination Complexes
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Boric acid (H3BO3, 99.97%), urea (NH2CONH2, 99.3+%), d -(+)-glucose (99%), calcium nitrate tetrahydrate (Ca(NO3)2·4H2O, 99%), magnesium nitrate hexahydrate (Mg(NO3)2·6H2O, 98.0–102.0%), strontium nitrate (Sr(NO3)2, 99.0+%), barium nitrate (Ba(NO3)2, 99.95%), silver nitrate (AgNO3, 99.9+%), nickel(ii ) nitrate hexahydrate (Ni(NO3)2·6H2O, 98%), 2,2′-bipyridine (99+%), lanthanum(iii ) oxide (La2O3, 99.99%) were purchased from Alfa Aesar. Cobalt nitrate hexahydrate (Co(NO3)2·6H2O, 99%) was purchased from Aladdin and cobalt chloride (CoCl2·6H2O, 99.99%) from MACKLIN.
Other analytical grade solvents including ethanol, acetonitrile and chemicals such as potassium hydroxide, ammonia solution, hydrochloric acid and triethanolamine were supplied by Sinopharm.
Other analytical grade solvents including ethanol, acetonitrile and chemicals such as potassium hydroxide, ammonia solution, hydrochloric acid and triethanolamine were supplied by Sinopharm.
4
Synthesis of Functional Polymers
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Di(ethylene glycol) methyl ether methacrylate (95%, D), copper (I) chloride (98%, CuCl), ethyl α-bromoisobutyrate (EBB, 98%), propargyl chloroformate (96%), 4-dimethylaminopyridine (99%, DMAP), 2-azido-1,3-dimethylimidazoliniumhexafluoro phosphate (97%, ADMP), and triethylamine (98%, TEA) were purchased from Sigma-Aldrich (Poznan, Poland). The 2-aminoethyl methacrylate hydrochloride (>98%, A) was received from Polysciences Inc. (Warrington, PA, USA). The 2,2′-bipyridine (99%, Bpy) was purchased from Alfa Aesar (Karlsruhe, Germany). Methyl alcohol (99.8%), ethyl alcohol (98%), dichloromethane (99.8%, DCM), N,N-dimethylformamide (DMF), and tetrahydrofuran (99.5%, THF) were purchased from Avantor™ Performance Materials (Gliwice, Poland). The DCM and DMF were distilled before use. The THF was distilled over potassium hydroxide before use. The water used to prepare polymer solutions was purified using a commercial ion exchange system (Hydrolab Company, Straszyn, Poland). The polymer solutions were kept overnight at 8 °C before use. Other reagents were used as received.
5
Copper-Mediated Oxidation of High-Purity hsDNA
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All chemicals were of analytical reagents and used as received without further purification. 2,2′-Bipyridine and AO were purchased from Alfa Aesar, and copper sulfate pentahydrate and other reagents were purchased from Merck. The hsDNA was purchased from Sigma Biological Co., its purity was monitored by the ratio of absorbance at 260–280 nm, and the ratio of 1.8–1.9 indicated that the hsDNA was free from protein. The hsDNA was dissolved in double-distilled water with 50 mmol·L−1 sodium chloride and dialyzed at 4°C for 48 h [38 (link)]. The hsDNA concentration was measured by UV-Vis at 260 nm. Tris-HCl buffer solution (pH 7.40) was prepared by using triple-distilled water.
6
Ruthenium(II) Complexes Synthesis and Characterization
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All materials were used as received without further purification unless otherwise noted. Diethyl ether, acetone, acetonitrile, and methanol were procured from Fisher Scientific, and dichloromethane was obtained from Macron Fine Chemicals. N,N-Dimethylformamide was purchased from Sigma-Aldrich and was dried under nitrogen by passing through a column of activated alumina. Reineke salt, ammonium hexafluorophosphate, lithium chloride, sodium chloride, sodium acetate trihydrate, ascorbic acid, 1,3-diphenylisobenzofuran (DPBF), acetylacetonate, 1,10-phenanthroline, 4,4′-dimethylbipyridyl, acetonitrile-d3 and acetone-d6 were purchased from Sigma-Aldrich, and 2,2′-bipyridine was obtained from Alfa Aesar. Sulfuric acid and sodium bicarbonate were received from Fisher Scientific, and potassium ferrioxalate was purchased from Strem Chemicals. Neutral alumina was purchased from Fisher Scientific. The starting material [Ru(p-cymene)Cl2]2,21 (link) and the complexes [Ru(p-cymene)(bpy)Cl]Cl, [Ru(p-cymene)(4,4′-Me2bpy)Cl]Cl, [Ru(p-cymene)(phen)Cl]Cl, and Ru(p-cymene)(acac)Cl were synthesized according to published procedures.21b
7
Preparation of Ruthenium and Cobalt Complexes
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All solvents and chemicals were of reagent-grade quality, obtained commercially and used without further purification, unless otherwise noted. Ruthenium(iii ) chloride hydrate, ammonium hexafluorophosphate, n-butyllithium solution (2.7 M in heptane), n-pentane, and tetrahydrofuran were purchased from Aldrich Chemicals. The chemicals, such as 4,7-dimethyl-1,10-phenanthroline (Me2phen), silver sulphate, lead dioxide, and cobalt(ii ) chloride, were purchased from Alfa Aesar. 2,2′-Bipyridine, 1,2,3,4,5-pentamethylcyclopentadiene, and TiIV(O)(tpyp) (tpyp = 5,10,15,20-tetra(4-pyridyl)porphyrinato anion) were obtained from Tokyo Chemical Industry Co., Ltd. Sc(NO3)3·4H2O was supplied by Mitsuwa Chemicals Co., Ltd. 18O2 gas (98% 18O-enriched) was purchased from ICON Services Inc. (Summit, NJ. USA). The purification of water (18.2 MΩ cm) was performed with a Milli-Q system (Millipore, Direct-Q 3 UV). Acetonitrile was dried according to published procedures and distilled prior to use.30 The cobalt(iii ) starting complex, [CoIII(Cp*)(bpy)(H2O)]2+ (1 , Cp* = η5-pentamethylcyclopenta-dienyl and bpy = 2,2-bipyridine), and the tris(4,7-dimethyl-1,10-phenanthroline)ruthenium(ii ) complex, [RuII(Me2phen)3]2+, were prepared according to the published methods.25 ,31
8
Synthesis and Characterization of Metal Complexes
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1,10‐phenanthroline (Sigma Aldrich, 97 %), 2–2’‐bipyridine (Alfa Aesar, 99 %), 4,7‐dimethyl‐1,10‐phenanthroline (Sigma Aldrich), 1,10‐phenanthroline‐5‐amine (Sigma Aldrich, 97 %), 4‐bromoaniline (Sigma Aldrich, 99 %), NaNO2 (Merck, 99.9 %), K3Fe(CN)6 (Sigma Aldrich, 99.98 %) and Cu(OTf)2 (Alfa Aesar, ≥99 %) were used as received. Electrolyte solutions were prepared with HClO4 (Merck suprapur, 70 %), HCl (Merck, 37 %), Na2HPO4 (Merck, 99.9 %), NaH2PO4 (Merck, 99.9 %), NaCl (Merck, 99.9 %) and were prepared with MilliQ water (>18.2 MΩ cm resistivity). Argon and oxygen (5.0) were purchased from Linde Gas.
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