Myrislignan ([erythro-(1R,2S)-2-(4-allyl-2,6-dimethoxyphenoxyl)-1-(4-hydroxy-3-methoxyphenyl) propan-1-ol], batch number DST180502-043) was purchased from Desite Biotechnology Co., Ltd. (Chengdu, China) and had a purity greater than 99%. For the in vitro experiments, myrislignan was dissolved in dimethyl sulfoxide (DMSO, Sigma, USA) to a concentration of 25 mg/ml and diluted in DMEM containing 1% FBS to different concentrations. Sulfadiazine (Sigma, USA), which was used as positive control drug, was dissolved in DMEM with 1% FBS to a concentration of 0.4 μg/ml. For the in vivo experiments, myrislignan was dissolved in solvent 1 (isotonic saline containing 5% ethanol and 5% Cremophor EL) to concentrations of 5 or 10 mg/ml. Sulfadiazine (10 mg/ml), pyrimethamine (5 mg/ml, Sigma, USA), and folinic acid (1.5 mg/ml, Sigma, USA) were suspended in physiological saline containing 1% CMC-Na (solvent 2) and used as a positive control.
Sulfadiazine
Manufactured by Merck Group
Sourced in United States, Germany, Brazil, United Kingdom
Sulfadiazine is a pharmaceutical compound used as a laboratory reagent. It is a white, crystalline solid with a melting point of approximately 252-254°C. Sulfadiazine is commonly used in various analytical and research applications in the field of chemistry and biology.
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Lab products found in correlation
Sulfamethoxazole, by Merck Group (19 mentions)
Sulfamethazine, by Merck Group (11 mentions)
Sulfamerazine, by Merck Group (10 mentions)
Sulfathiazole, by Merck Group (9 mentions)
Trimethoprim, by Merck Group (8 mentions)
Sulfapyridine, by Merck Group (8 mentions)
Pyrimethamine, by Merck Group (7 mentions)
Tetracycline, by Merck Group (6 mentions)
Enrofloxacin, by Merck Group (6 mentions)
Sulfadimethoxine, by Merck Group (6 mentions)
Erythromycin, by Merck Group (6 mentions)
Sulfachloropyridazine, by Merck Group (6 mentions)
Oxytetracycline, by Merck Group (5 mentions)
Chloramphenicol, by Merck Group (5 mentions)
Sulfadoxine, by Merck Group (5 mentions)
Amoxicillin, by Merck Group (5 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (4 mentions)
Azithromycin, by Merck Group (4 mentions)
Norfloxacin, by Merck Group (4 mentions)
Ciprofloxacin, by Merck Group (4 mentions)
68 protocols using sulfadiazine
1
Myrislignan Bioactive Compound Evaluation
2
Drug Preparation for Villous Explant Treatment
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Azithromycin (Biofarma, Uberlândia, MG, Brazil), spiramycin (Sigma) and a drug combination (PS) consisting of pyrimethamine (Sigma) and sulfadiazine (Sigma) were dissolved in DMSO (stock solution) to a concentration of 10,000 μg/mL for Azithromycin, spiramycin, sulfadiazine and 3000 μg/mL for pyrimethamine. Stock solutions were freshly reconstituted and different drug concentrations were used for the treatment of villous explants.
3
Auranofin, Pyrimethamine, and Sulfadiazine Evaluation
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Auranofin (Enzo Life Sciences), was dissolved in 100% ethanol as a stock solution (4 mg/mL) and then diluted in complete tissue culture medium (DMEM +2% FBS) for final concentrations of 0.1 to 19 µM. For in vivo experiments, the Auranofin concentration used was 1 mg/kg of estimated body weight.
Pyrimethamine (Sigma Aldrich) was dissolved in 100% ethanol in a stock concentration of 5 mg/mL. Three final dilutions in complete tissue culture medium were examined: 0.02, 0.1 and 0.2 µM. Sulfadiazine (Sigma Aldrich) was dissolved in complete medium at a final stock concentration of 5 mg/mL. Three doses in complete medium were evaluated: 0.2, 1 and 2 µM. Testing of Pyrimethamine/Sulfadiazine combinations by checkerboard method (using abovementioned concentrations) was carried out in triplicates and in three independent experiments.
Pyrimethamine (Sigma Aldrich) was dissolved in 100% ethanol in a stock concentration of 5 mg/mL. Three final dilutions in complete tissue culture medium were examined: 0.02, 0.1 and 0.2 µM. Sulfadiazine (Sigma Aldrich) was dissolved in complete medium at a final stock concentration of 5 mg/mL. Three doses in complete medium were evaluated: 0.2, 1 and 2 µM. Testing of Pyrimethamine/Sulfadiazine combinations by checkerboard method (using abovementioned concentrations) was carried out in triplicates and in three independent experiments.
4
Dilution Preparation of Antimicrobial Compounds
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Compounds were dissolved in dimethyl sulfoxide (DMSO, Sigma-Aldrich, St. Louis, MO, USA) to 125 mM. The final concentration of DMSO in the compounds dilutions was not higher than 1.00%. Drug, sulfadiazine [4-amino-N-(2-pyrimidinyl)benzenesulfonamide] (S8626, Sigma-Aldrich) was dissolved in 1M sodium hydroxide (NaOH, Sigma-Aldrich) to 400 mM. Final concertation of NaOH in sulfadiazine dilutions was not higher than 2.5%. Ready solution sulfadiazine and trimethoprim in weight ratio 5:1 (Sul-Tridin 24%, 200 mg/mL [800 mM] + 40 mg/mL [137.78 mM]– ScanVet Poland). Dilution of the all compounds and drugs were freshly prepared before the cells were exposed in appropriate medium.
5
Quantification of Veterinary Antibiotics in Samples
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Standards of five veterinary antibiotics (VAs) (Figure 3 ) (Tetracycline (TC, 90%), OxyTetracycline (OTC, 98%), Sulfadiazine (SDZ, 98%), Norfloxacin (Norf, 98%), Enrofloxacin (Enorf, 98%)) were purchased from Sigma-Aldrich (St. Louis, MO, US) (CAS Reg. No. 60-54-8 for TC, CAS Reg. No. 6153-64-6 for OTC, CAS Reg. No. 68-35-9 for SDZ, CAS Reg. No. 70458-96-7 for Norf, and CAS Reg. No. 93106-60-6 for Enorf) (the structures are presented in Figure 4 ). Acetonitrile (ACN) and methanol (MeOH) of HPLC grade were purchased from Sigma-Aldrich. Oxalic acid, disodium ethylene–diaminetetraacetat (Na2EDTA), disodium hydrogen phosphate (Na2HPO4), and sodium dihydrogen phosphate (NaH2PO4) of analytical-reagent grade were obtained from Beijing Chemicals Company (Beijing, China). Deionized water (Milli-QMillipore, Bedford, MA, USA) was used in the study.
6
Compound Solubilization for Cell Assays
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Compounds 2 –22 were dissolved in dimethyl sulfoxide (DMSO—Sigma) to 50,000 μg/mL. The final concentration of DMSO in compounds dilutions was not higher than 1.00%. sulfadiazine [4-amino-N-(2-pyrimidinyl)benzenesulfonamide] (S8626, Sigma) was dissolved in 1 M sodium hydroxide (NaOH—Sigma) to 100 mg/mL. Final concertation of NaOH in sulfadiazine dilutions was not higher than 2.5%. Dilution of the all compounds 2 –22 and drug were freshly prepared before the cells were exposed.
7
Quantification of Sulfadiazine and Sulfamerazine
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Sulfadiazine and sulfamerazine were purchased from Sigma-Aldrich (Steinheim, Germany) and were of the highest purity available. Methanol (HPLC-grade) was obtained from Applichem, Darmstadt, Germany.
8
Purification of Malaria Gametocytes
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Purification of gametocytes was achieved using a protocol modified from7 (link). Mice were treated i.p with 200 μl phenylhydrazinium chloride (PH; 6 mg/ml in PBS; ProLabo UK). Day four post infection (p.i.) mice were treated with sulfadiazine (Sigma) at 20 mg/L in their drinking water for two days to eliminate asexual blood stage parasites. On day six p.i. mice were bled by cardiac puncture into heparin and gametocytes separated from uninfected erythrocytes on a 48% NycoDenz gradient (27.6% w/v NycoDenz in 5 mM Tris-HCl, pH 7.20, 3 mM KCl, 0.3 mM EDTA) in coelenterazine loading buffer (CLB), containing PBS, 20 mM HEPES, 20 mM Glucose, 4 mM sodium bicarbonate, 1 mM EGTA, 0.1% w/v bovine serum albumin, pH 7.25. Gametocytes were harvested from the interface and washed twice in RPMI 1640 ready for activation of gamete formation.
9
Chronic Toxoplasma Infection in Mice
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This experimental study and all administrations were reviewed and approved by the Ethical Committee of Animal Experiments at Sun Yat-sen University.
Female 6- to 8-week-old C57BL/6 and BALB/c mice were purchased from the Experimental Animal Center at Sun Yat-sen University (Guangzhou, China), and 20 mice were used per each group. All animals were housed under specific-pathogen-free conditions in the animal facility at Sun Yat-sen University. Mice were infected via oral route with eight cysts of T. gondii Pru strain prepared from the brain of chronically infected mice. To establish a chronic infection by controlling the proliferation of tachyzoites during acute stage, mice were treated with sulfadiazine (Sigma-Aldrich, Shanghai, China) in the drinking water as described previously (23 (link)).
Female 6- to 8-week-old C57BL/6 and BALB/c mice were purchased from the Experimental Animal Center at Sun Yat-sen University (Guangzhou, China), and 20 mice were used per each group. All animals were housed under specific-pathogen-free conditions in the animal facility at Sun Yat-sen University. Mice were infected via oral route with eight cysts of T. gondii Pru strain prepared from the brain of chronically infected mice. To establish a chronic infection by controlling the proliferation of tachyzoites during acute stage, mice were treated with sulfadiazine (Sigma-Aldrich, Shanghai, China) in the drinking water as described previously (23 (link)).
10
Multiplex Antimicrobial Residue Detection
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Chloramphenicol (CAP), ciprofloxacin (CIP), florfenicol, (FF), penicillin (PEN), ractopamine (RAC), salbutamol (SAL), sulfadiazine (SUL), and thiamphenicol (TAP) standards were purchased from Sigma Aldrich (99% purity, St. Louis, MO, USA). CAP-conjugated antigen and HRP-labeled CAP antibody were obtained from ZeYang Co. (Beijing, China). SuperSignal chemiluminescence substrate solution was purchased from Thermo Fisher (USA). Milli-Q Synthesis system was obtained from Millipore (Bedford, MA, USA) for water purification. Other reagents (analytical grade) were purchased from Beijing Reagent Corp. (Beijing, China).
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