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155 protocols using allopurinol

1

Antioxidant and Uric Acid-Lowering Effects

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BHA and allopurinol were purchased from Sigma-Aldrich. BHA was dissolved in dimethyl sulfoxide (DMSO) and stored at -20°C until use. BHA was orally administered at 200 mg/kg/day using a sonde needle from 2 to 5 dpi. According to the Material Safety Data Sheet (MSDS) (ScienceLab. com, Inc. Dickinson, Texas), the oral LD50 of BHA is 1,100 mg/kg in mice. allopurinol was prepared in PBS and then stored at -20°C until use. allopurinol was orally administered at 2 mg/kg/day using a sonde needle from -1 to 14 dpi. According to the MSDS (Sigma-Aldrich), the oral LD50 of allopurinol is 78 mg/kg in mice.
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2

Allopurinol Supplemented Fly Food Preparation

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Allopurinol stock solution (300 mM) was made by dissolving 0.41 g of Allopurinol (Sigma-Aldrich, cat# A8003-25g) in 10 ml of 1M sodium hydroxide. Normal cornmeal molasses food was melted and supplemented with 1/100 vol of Allopurinol stock solution. After mixing well, the Allopurinol food was poured into individual vials. Female flies with appropriate genotypes were cultured on Allopurinol food as indicated in figure legends.
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3

Renal Ischemia-Reperfusion Injury Model

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Briefly, rats were anesthetized by intraperitoneal injection with pentobarbital (50 mg/kg). Rats were placed on an electric heating pad to maintain a body temperature of 37°C. A right nephrectomy was performed. The left renal artery and vein were blocked using a vascular clamp for 30 min to induce ischemia and the left kidney was reperfused for 72 h. Finally, all rats were killed.
Rats were separated into the following three groups. In Group 1, sham operated control rats (n=8) were subjected to right nephrectomy, but without the induction of left renal ischemia. In Group 2, ischemia/reperfusion (I/R) rats (n=8) were subjected to right nephrectomy and left renal ischemia for 30 min, followed by a 72 h reperfusion period. In Group 3, rats treated with allopurinol preconditioning + I/R (n=8), prior to I/R manipulation (as in group 2), were administered allopurinol 50 mg•kg -1 •day -1 (Sigma, St. Louis, MO, USA) IP 15 (link) for two weeks, as described previously 2 (link) , and then were subjected to the same manipulations as group 2.
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4

Preparation of Allopurinol and AFPR Solutions

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The allopurinol (Sigma-Aldrich, St. Louis, Missouri, USA) solution, AFPR solutions and potassium oxonate (Sigma-Aldrich, St. Louis, Missouri, USA) suspension were prepared according to the medium weight of each group. allopurinol and AFPR were solubilized in Tween-80: water (2: 98) (vehicle) (Sigma-Aldrich, St. Louis, Missouri, USA). potassium oxonate was prepared in a suspension with NaCl 0.9% solution.
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5

Polarization of Bone Marrow-Derived Macrophages

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BMMФ were stimulated either with IL-4 (10 ng/ml, AF-214-14, Peprotech), lipopolysaccharide (100 ng/ml, Sigma) or dexamethasone (100nM, D4902, Sigma) to respectively achieve M(IL-4), M(LPS) or M(GC) polarization state. In some experiments, BMMФ were treated with C75 (Sigma), Cerulenin (Sigma), Etomoxir (Sigma), pan-AKT inhibitor MK-2206 (1 mM, Cayman Chemical), 25-hydroxycholesterol (10 μM, Cayman chemical), N-acetyl cysteine (10mM, Sigma), Diphenyleneiodonium chloride (DPI, 5 μM, Sigma), L-NG-Nitroarginine methyl ester (L-NAME, 1mM, Cayman Chemical), Allopurinol (100 μM, Sigma), hydrogen peroxide (Sigma), SIRT1 activator II (Sigma), EX-527 (Sigma), Compound C (Sigma), AICAR (Abcam), HMGCoA (Sigma), water-soluble cholesterol (Cholesterol–methyl-β-cyclodextrin, Sigma) or Simvastatin (Sigma).
For fatty acid treatment, palmitic or oleic acid (Cayman) were solubilized at 100mM in absolute ethanol at 60°C. Fatty acid were conjugated at the desired concentration using a sonicator water bath into the macrophage culture medium to avoid endotoxin contamination from BSA54 (link).
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6

Antioxidant and Antibacterial Evaluation

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Methanol, hexane, chloroform, ethyl acetate and ethanol were purchased from Junsei Chemical Co., Ltd., Tokyo, Japan. Potassium phosphate monobasic and dibasic, xanthine, xanthine oxidase, allopurinol, and hydrochloric acid were obtained from Sigma-Aldrich Corp., St. Louis, MO, USA. Reagents including 1,1-diphenyl-2-picrylhydrazyl (DPPH), sodium acetate, acetic acid, 2,2′-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), potassium peroxodisulfate, and dibutyl hydroxytoluene (BHT) were supplied by Kanto Chemical Co. Inc., Tokyo, Japan. Four bacteria including Staphylococcus aureus, Escherichia coli, Bacillus subtilis, and Proteus mirabilis were provided by Sigma-Aldrich Corp., St. Louis, MO USA. All chemicals used were of analytical grade.
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7

Aristolochia bracteolata Leaf Phytochemical Analysis

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Aristolochia bracteolata leaves were procured from authorized supplier and identified using literature provided. All the chemicals of analytical grade were used. Petroleum ether, ethanol, ethylacetate, chloroform and acetone were procured from Nanjing Hanbang Chemical Reagent Company (Nanjing, PR China). Allopurinol, XOD and potassium oxonate were purchased from Sigma Aldrich (USA). From the local laboratory chemical suppliers, authentic kits for quantification of biochemical markers such as uric acid, BUN and creatinine were brought. All other chemicals used were of analytical grade and purchased from Sigma (USA).
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8

Antioxidant Assay Protocol

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BPA, catalase, ebselen, LiCl2, allopurinol, and glutathione‐ethyl ester, and MnTMPyP were purchased from Sigma (St Louis, MO); MnTBAP was purchased from Biomol (Butler Pike, PA); L‐NAME was purchased from Tocris (Ellisville, MO); and cell culture ingredients were obtained from Gibco BRL (Gaithersburg, MD).
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9

Vascular Reactivity Protocol

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L-NA hydrochloride, ACh chloride, diethylamine NONOate diethylammonium salt, CGRP (8–37), TTX, L-NAME hydrochloride, 7-NI, 1400W, phentolamine, apocinin, allopurinol, lucigenin, tiron, tempol and DAF-2 (Sigma-Aldrich, Madrid, Spain) were used. Stock solutions (10 mmol/L) of drugs were made in distilled water, except for NA, which was dissolved in a NaCl (0.9%)-ascorbic acid (0.01% w/v) solution. These solutions were kept at -20°C and appropriate dilutions were made in KHS on the day of the experiment.
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10

Enzymatic Biocatalysis with Immobilized CALB

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CALB immobilised in acrylic resin (EC 3.1.1.3, 10,000 U/g) (Novozyme 435), xanthine oxidase from bovine milk, xanthine, allopurinol, hesperidin, naringin, and rutin standards and hexanoic, octanoic, decanoic, lauric, and oleic acids were obtained from the Sigma-Aldrich Chemical Co., St. Louis, MO. All the solvents and reagents were of analytical, spectrometric, or chromatographic quality.
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