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4 protocols using buffer solutions

1

Photocatalytic Oxidation Process Characterization

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Melamine 99% was purchased from Alfa Aesar, UK. Potassium persulfate (99%) was obtained from SD fine chemicals, India. AAP (98%), tert-Butanol (99%), and 1,4-Benzoquinone (98%) were purchased from Loba Chemie Pvt. Ltd, India. Ethylenediamine tetraacetic acid disodium hydrate (99%) was purchased from Himedia, India. Sodium azide 99% was obtained from s d fine chemicals. Buffer solutions (pH 4.00, 7.00, and 10.01) were procured from Thermo Fischer Scientific, India. Methanol, Acetonitrile and Formic acid, were of HPLC grade and purchased from FINAR chemicals, India. Sodium sulfate (99.5%) is used as an electrolyte for photocurrent measurement and was bought from Qualigens, India. Dimethyl-1-pyrroline-N-oxide (DMPO) and 2,2,6,6-tetramethylpiperidine (TEMP) were purchased from Tokyo Chemical Industry Co., Ltd. (TCI), India. All the solutions were prepared using RO water (conductivity 4.0–5.4 μS/cm).
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2

Synthesis and Characterization of Glycerol Monomethacrylate Copolymers

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Glycerol monomethacrylate (GMA; 99.8%) was donated by GEO Specialty Chemicals (Hythe, UK) and was used without further purification. 2-Hydroxypropyl methacrylate (HPMA) was purchased from Alfa Aesar (UK) and was used as received. 4,4′-Azobis(4-cyanopentanoic acid) (ACVA; V-501; 99%), 2,2′-azobisisobutyronitrile (AIBN), 2-cyano-2-propyl dithiobenzoate (CPDB), 4-carboxyphenylboronic acid (CPBA; ≥90%), ethanol (99%, anhydrous grade), methanol, and dichloromethane were purchased from Sigma-Aldrich (UK) and were used as received. Deuterated methanol (CD3OD) was purchased from Goss Scientific (Nantwich, UK). Buffer solutions were purchased from Thermo Fisher Scientific (Chelmsford, USA). All solvents were HPLC-grade and were purchased from Fisher Scientific (Loughborough, UK).
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3

Cell Cytotoxicity Assay Protocol

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All chemicals and reagents were used as received from Sigma-Aldrich without further purification unless otherwise specified. Minimum essential medium (MEM), Dulbecco’s modified Eagle’s medium (DMEM), phosphate buffered saline (PBS), fetal bovine serum (FBS), penicillin and streptomycin, LysoTracker® Red DND-99, and (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) were purchased from Invitrogen. Buffer solutions (pH = 1–13) were purchased from Fisher Scientific. Chloroquine was purchased from Bide Pharmatech Ltd. Milli-Q water was supplied by Milli-Q Plus System (Millipore Corporation, United States).
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4

Assessing Bacterial Impact on Cell Viability

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Cell viability was determined by means of trypan blue dye enumeration. The cells were exposed to one of the six selected bacterial strains under anaerobic conditions for 24 h. After staining with trypan blue, viable cells were counted using an automated cell counter (Countess Automated Cell Counter, Invitrogen, UK). In addition, the pH was measured by collecting conditioned media after SCAP was cocultured with bacteria for 24 h. Before starting measurements, the pH meter (Mettler-Toledo, Columbus, OH, US) was calibrated using buffer solutions at pH 4.0, 7.0, and 9.21 (Fisher Scientific).
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