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10 protocols using trans chalcone

1

Trichophyton rubrum Growth Conditions

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Trichophyton rubrum CBS 118892 was cultured on Sabouraud dextrose agar (Oxoid, Hampshire, England) at 28 °C, as described previously [28 (link)]. Conidial suspensions were obtained from 15-day-old plates. The conidial concentration was determined in a Neubauer chamber and approximately 1.6 × 106 conidia were added to 20 mL of liquid Sabouraud and incubated for 72 h at 28 °C under shaking at 150 rpm. The resulting mycelia were incubated under six different conditions: i) control medium (MMNG): Cove’s minimal medium [29 (link)] containing 70 mM nitrate (Sigma Aldrich, St. Louis, MO, USA) and 50 mM glucose (Sigma Aldrich); ii) keratin medium (MMK): Cove’s minimal medium supplemented with 0.5% bovine keratin; iii) elastin medium (MME): Cove’s medium supplemented with 0.25% elastin (Sigma Aldrich); iv) MMNG+TChal: MMNG medium containing 0.24 μg/mL of trans-chalcone (Sigma Aldrich); v) MMK + TChal: MMK containing 0.24 μg/mL of trans-chalcone, and vi) MME + TChal: MME containing 0.24 μg/mL of trans-chalcone. The pH of the medium was 5.0 in all conditions and the cultures were incubated for 3, 7, and 14 days at 28 °C under shaking (130 rpm). The concentration of trans-chalcone was based on its minimal inhibitory concentration as reported previously [9 (link)].
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2

Compound Sources for Biological Assays

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The compounds dimethyl fumarate (DMF), trans-chalcone, curcumin, sulforaphane, tert-butylhydroquinone (tBHQ) and dimethyl itaconate (DMI) were purchased from Sigma-Aldrich. 4-OI was chemically synthetized as described elsewhere [9 (link)]. The prostaglandins 15d-PGJ2, PGE2 and PGD2 were purchased from Cayman Chemicals, while epoxycyclopentenone (EC), cyclo-epoxycyclopentenone (cEC), EC-reduced were synthetized as previously described [[31] (link), [32] (link), [33] (link)]. The antioxidant N-acetylcysteine (NAC) and the pan-caspase inhibitor Q-VD-OPh (QVD) were purchased from Sigma-Aldrich, whereas the necroptosis inhibitor Nec-1s was from Merck Millipore.
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3

Synthesis and Characterization of 4'-Hydroxychalcone

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4′‐Hydroxychalcone was obtained from the internal compound collection of RBG, Kew. Purity and structure were verified using LC–MS and NMR. trans‐Chalcone was purchased from Sigma‐Aldrich, Gillingham, UK.
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4

HPLC-DAD Analysis of Phytochemicals

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Solvents for the HPLC-DAD analyses were of analytical grade and were purchased from Sigma (St. Louis, MO, USA). Water was purified by a Milli-Q Plus system from Millipore (Milford, MA, USA). The following commercial compounds were used as standards: Gallic acid, pyrogallol, 4-hydroxy benzhydrazide, epigallocatechin, vanillic acid, (+)-catechin hydrate, syringic acid, phloroglucinol, p-anisic acid, dexamethasone, chlorogenic acid, caffeic acid, p-coumaric acid, trans-ferulic acid, naringin, 2-amino-3,4-dimethyl benzoic acid, coumarin, morin hydrate, luteolin, hesperetin, alizarin, biotin, trans-chalcone, rutin hydrate, myricetin, quercetin, rhein, 3-hydroxyflavone, and emodin were purchased from Sigma–Aldrich Co. Catechin gallate and protocatechuic acid were obtained from Santa Cruz Biotech (Santa Cruz, CA, USA). All other chemicals were analytical grade.
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5

Amyloid Aggregation Inhibitor Screening

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Silibinin, trans-chalcone (benzylideneacetophenone), thioflavin S, and Congo red were purchased from Sigma (USA). Regular insulin was purchased from EXIR Pharmaceutical Company (Iran). Chloroform, monopotassium phosphate (KH2PO4), potassium phosphate (KH2PO), and dimethyl sulfoxide (DMSO) were purchased from Merck (Germany).
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6

Chalcone and ER Stress Modulation

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trans-Chalcone, N-acetyl cysteine (NAC), tunicamycin (Tu), thapsigargin (Tg), rotenone, thallium trifluoroacetate, allopurinol, mefenamic acid, apocynin, n-ethylmalemide, iodacetamide were purchased from Sigma Aldrich (St Louis, MO, USA). Z-VAD-fmk was purchased from Enzo-life Technologies (Farmingdale, NY, USA). GKT137831 was obtained from Cayman Chemical (Michigan, USA). Hoechst 33342 and ER-Tracker™ Green were purchased from Invitrogen (Invitrogen, CA, USA).
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7

Antifungal Compound Evaluation Protocol

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Trans-chalcone and α-solanine were purchased from Sigma-Aldrich (St. Louis, MO, USA) and diluted in 10% aqueous DMSO. Terbinafine was purchased from Sigma-Aldrich (St. Louis, MO, USA) and diluted in 5% aqueous DMSO. The final concentration of DMSO used in the antifungal and coculture assays was fixed at a maximum of 0.5%. Terbinafine was used as positive control (commercial antifungal involved in ergosterol synthesis inhibition) [9 (link)]. Negative controls consisted of DMSO without the tested compounds at a final concentration of 0.5%.
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8

Polymer Blends with Natural Antioxidants

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The object of the research was to study polymer blends based on polylactide (PLA) and polyhydroxybutyrate (P(3.4 HB) contained selected natural antioxidants. Polylactide (PLA), Ingeo 4043D, was obtained from NatureWorks LLC (Minnetonka, MN, USA). This material with a density of 1.25 g/cm3 is characterized by glass transition of about 55–60 °C. The melting point of this polymer is 145–160 °C and the melt flow index equals 6 g/10 min. Poly(hydroxybutyrate) (P(3.4 HB)) containing 12 mol% 4-hydroxybutyrate was obtained from Simag Holdings LTD (Hong Kong, China). The density of this polymer is 1.25 g/cm3, its melt flow index equals 18 g/10 min and the melting point is 170 °C. Different natural antioxidants, including lignin (alkali, low sulfonate content lignin, kraft, pH: 10.5 (3 wt %)), flavone (Mw = 222.24 g/mol, mp: 94–97 °C, ≥99.0%) and trans-chalcone (Mw = 208.26 g/mol, mp: 55–57 °C, 97%), used as stabilizers, were obtained from Sigma-Aldrich (Saint Louis, MI, USA).
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9

Anticancer Effects of Chalcone Compounds

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The two tested compounds, trans-chalcone and Licochalcone A, and some of the main reagents including trypsin, doxorubicin, hydrochloride, dimethyl sulfoxide (DMSO), DMEM, penicillin, kanamycin, and streptomycin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Fetal bovine serum was obtained from Cultilab (Campinas, SP, Brazil). The materials used for gene expression were RNeasy® Mini Kit, RNase-free DNase Set, RT2 SYBR® Green JumpStart TAQ Ready Mix, RT2 First Strand Kit, and RT² Profiler™ PCR Array Human Cancer Drug Targets. The BCA (Bicinchoninic Acid) protein assay was purchased from Thermo Scientific (Rockford, IL, USA).
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10

Chalcone-Induced Analgesic and Anti-Inflammatory Assay

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Trans-Chalcone (Sigma-Aldrich), diclofenac sodium, complete
Freund’s adjuvant, formalin, DMSO, chloroform, isopropanol,
and ethanol were from Sigma-Aldrich, besides TRI-Reagent RT (BioShop
Canada), WizScript cDNA Synthesis Kit (High Capacity) (Wizbiosolutions),
GI I EvaGreen qPCR Master Mix (GeneDirex), Rat PGE2 ELISA Kit (Wuhan
Zokeyo Biotechnology Co., Ltd.), and nitric oxide (NO) assay kit (Solarbio
Life Sciences).
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