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3 protocols using methyl 3 mercaptopropionate

1

Synthesis and Characterization of Polymeric Antimicrobials

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Ethanolamine, 4-amino-1-butanol, di-tert-butyl dicarbonate, triethylamine, methyl 3-mercaptopropionate, and ethyl methacrylate were purchased from Acros Organics. 2,2’-azobisisobutyronitrile (AIBN) and 2-(trimethylsiloxy)ethyl methacrylate were purchased from Sigma-Aldrich Co. LLC. Trifluoroacetic acid (TFA) and solvents were purchased from Thermo Fisher Scientific, Inc. The chemicals were used without further purification, with the exception of methacryloyl chloride, which were distilled before use. 1H NMR was performed using a Varian MR400 (400 MHz) and analyzed using VNMRJ 3.2 and MestReNova. Gel permeation chromatography (GPC) analysis was performed using a Waters 1515 HPLC instrument equipped with Waters Styragel (7.8 × 300 mm) HR 0.5, HR 1, and HR 4 columns in sequence and detected by a differential refractometer (RI). Mueller Hinton broth (MHB, BD and Company ©) and phosphate buffered saline (PBS, pH=7.4, Gibco®) were prepared according to manufacturer instructions and sterilized prior to use. Human red blood cells (RBCs) (leukocytes reduced adenine saline added) were obtained from the American Red Cross Blood Services Southeastern Michigan Region and used prior to the out date indicated on each unit.
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2

Synthesis and Characterization of Functionalized Monomers

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tert-Butyl-3-hydroxypropanoate, methyl 3-mercaptopropionate, and ethyl methacrylate were purchased from Acros Organics. 2,2′-Azobis-(isobutyronitrile) (AIBN) and ethyl methacrylate were purchased from Sigma-Aldrich Co. Trifluoroacetic acid (TFA) and solvents were purchased from Thermo Fisher Scientific, Inc. The chemicals were used without further purification, with the exception of methacryloyl chloride which was distilled before use and AIBN which was recrystallized from hot methanol. Boc-amino ethyl methacrylate was synthesized according to the previous reported procedure.61 (link)1H NMR was performed using a Varian MR400 (400 MHz), 13C NMR was performed using a Bruker NEO600 (600 MHz) and the data was analyzed using VNMRJ 3.2 and MestReNova. Gel permeation chromatography (GPC) analysis was performed using a Waters 1515 HPLC instrument equipped with Waters Styragel (7.8 × 300 mm) HR 0.5, HR 1, and HR 4 columns in sequence and detected by a differential refractometer (RI). Mueller Hinton broth (MHB, BD and Company) and phosphate buffered saline (PBS, pH = 7.4, Gibco) were prepared according to manufacturer instructions and sterilized prior to use. Human red blood cells (RBCs) (leukocytes reduced adenine saline added) were obtained from the American Red Cross Blood Services Southeastern Michigan Region and used prior to the out date indicated on each unit.
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3

MRSA Colonies Isolation and Analysis

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Twelve MRSA colonies (including three VISA colonies containing the mecA gene) which were isolated from the blood of ten patients treated for blood stream infections at the University of Michigan Health System were used (Table 1). Human adult fibroblasts (PCS-201-012) and fibroblast basal medium (ATCC PCS-201-030) were obtained from ATCC (Manassas, VA, USA). Muller-Hinton (MH) broth was purchased from Fisher Scientific (Pittsburg, PA, USA). Vancomycin hydrochloride was purchased from Hopira, Inc. (Chicago, IL, USA). 96-well plates were purchased from Corning (Constar 3591, Corning, NY, USA). Cell counting kit-8 (CCK-8) was purchased from Dojindo laboratories (Kumamoto, Japan). 4-Amino-1-butanol was purchased from TCI Chemicals. Di-tert-butyl dicarbonate was purchased from Oakwood Chemicals. 2,2-azobisisobutyronitrile (AIBN) was purchased from Sigma-Aldrich (St Louis, MO). 2-cyanoprop-2-yl-dithiobenzoate was purchased from Strem Chemicals (Newburyport, MA). Trifluoracetic acid (TFA) and the solvents hexanes, dichloromethane, diethyl ether and methanol were purchased from Fisher Scientific. Ethyl methacrylate (EMA), methacryloyl chloride, 4-butanolamine, methyl 3-mercaptopropionate and di-tert-butyldicarbonate were purchased from Acros Organics (part of Thermo Fisher Scientific, Geel, Belgium).
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