The Candida spp. reference strains used in this study were kindly provided by Fundação Oswaldo Cruz (FIOCRUZ, Rio de Janeiro, Brazil) and included four C. albicans (IOC 2508, IOC 2517, IOC 3703, and IOC 3704), C. parapsilosis (CP310), C. tropicalis (CT 309), and C. glabrata (CG74) strains. Sabouraud dextrose agar (SDA) (HIMEDIA, Mumbai, India) was used for fungal growth. RPMI 1640 medium supplemented with sodium bicarbonate and glutamine (Thermo Fisher, Waltham, MA, USA), 2% dextrose (Merck, Darmstadt, Germany), 0.165 mol/L 3-(N-morpholino) propanesulfonic acid, 4-morpholine-propanesulfonic acid (MOPS) (Thermo Fisher, Waltham, MA, USA), and 0.2% chloramphenicol was used in the susceptibility tests. The commercial antifungals used in this study were fluconazole, ketoconazole, nystatin (Infinity Pharma, Campinas, Brazil), and amphotericin B (Cristália, São Paulo, Brazil).
Amphotericin b
Manufactured by Cristália
Sourced in Brazil
Amphotericin B is a laboratory-grade antifungal agent. It is used in research and development settings for the examination and study of fungal organisms.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Merck Group (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
4 morpholine propanesulfonic acid mops, by Thermo Fisher Scientific (1 mentions)
Dextrose, by Merck Group (1 mentions)
Zymosan, by Merck Group (1 mentions)
Neutral red, by Merck Group (1 mentions)
Schneider s medium, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin, by Merck Group (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
Griess reagent, by Merck Group (1 mentions)
Ethanol, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Formaldehyde, by Merck Group (1 mentions)
Glacial acetic acid, by Merck Group (1 mentions)
Anhydrous sodium sulfate, by Merck Group (1 mentions)
Pf 429242, by Merck Group (1 mentions)
Miltefosine, by Cayman Chemical (1 mentions)
Antimycin a, by Merck Group (1 mentions)
Penicillin g, by Merck Group (1 mentions)
4 protocols using amphotericin b
1
Antifungal Susceptibility Testing of Candida spp.
2
Cytotoxicity and Nitric Oxide Assay
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Dimethyl sulfoxide (DMSO: 99%), anhydrous sodium sulfate, glacial acetic acid, ethanol, formaldehyde, sodium chloride, calcium acetate, zymosan, and neutral red were purchased from Merck Chemical Company (Germany). Schneider's medium, RPMI 1640 medium, fetal bovine serum (FBS), MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide), Griess reagent (1% sulfanilamide in H3PO4 10% (v/v) in Milli-Q water), and the antibiotics penicillin and streptomycin were purchased from Sigma Chemical (St. Louis, MO, USA). The antibiotic amphotericin B (90%) was purchased from Cristália (São Paulo, SP, Brazil).
3
Antileishmanial Compound Evaluation
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The compound tested in this work, PF-429242, was purchased from Sigma-Aldrich and diluted in deionized water. Amphotericin B, obtained from Cristália, was used as a reference drug in antileishmanial tests against promastigote and amastigote forms, it was also diluted with water and the following concentrations were used in the tests: 0.5, 0.25, 0.12, 0.06, 0.03, and 0.01 μM. Antimycin A was purchased from Sigma-Aldrich and used at 10 μM. Miltefosine was obtained from Cayman Chemical and used at 8 μM.
4
Bladder Cancer Cell Lines Treated with Gemcitabine
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Two urothelial bladder cancer cell lines, acquired from the Cell Bank of Rio de Janeiro, Brazil, were used in the in vitro experiments: i. J82, J82, derived from high-grade (grade III) tumor with three point mutations in the TP53 gene (two in exon 8 (codon 271: Glu (GAG) → Lys (AAG); codon 274 Val (GTT) → Phe (TTT)) and one in exon 9 (codon 320 Lys (AAG) → Asn (AAC)) [41 (link),42 (link),43 (link)]; ii. UM-UC-3, derived from a high-grade tumor with a point mutation at TP53 (exon 4, codon 113: Phe (TTG) → Cys (TCG)) [41 (link),42 (link),43 (link)].
Both cell lines were cultivated in monolayers, using DMEM culture medium supplemented with 10% bovine serum and 100 U/mL of penicillin G, 100 U/mL of streptomycin (Sigma-Aldrich, Saint Louis, EUA), and 2.5 µg/mL of amphotericin B (Cristália, Itapira, Brazil), and maintained at 37 °C in an atmosphere of 5% CO2. All experiments were performed with exponentially growing cells [37 (link)]. Gemcitabine (Gemzar) was obtained from Eli Lilly do Brazil Ltd.a. Treatment with gemcitabine was set to 24 h. Dilutions were performed in nuclease-free water.
Both cell lines were cultivated in monolayers, using DMEM culture medium supplemented with 10% bovine serum and 100 U/mL of penicillin G, 100 U/mL of streptomycin (Sigma-Aldrich, Saint Louis, EUA), and 2.5 µg/mL of amphotericin B (Cristália, Itapira, Brazil), and maintained at 37 °C in an atmosphere of 5% CO2. All experiments were performed with exponentially growing cells [37 (link)]. Gemcitabine (Gemzar) was obtained from Eli Lilly do Brazil Ltd.a. Treatment with gemcitabine was set to 24 h. Dilutions were performed in nuclease-free water.
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