Fluorochrome labels were administered via subcutaneous injection 1 week prior to surgery (xylenol orange tetrasodium salt, 100 mg/kg, Sigma-Aldrich, St. Louis, MO) and 4 days prior to euthanasia (alizarin complexone, 18 mg/kg, Sigma-Aldrich, St. Louis, MO). Following euthanasia, tibias were harvested with surrounding soft tissue and fixed for 48 hours in 4% formaldehyde in Dulbecco’s PBS (with Ca2+, Mg2+). Micro-CT scans (55 kV, 145 mA, 200 ms integration time, µCT 40, Scanco, Brüttisellen, Switzwerland) with 8 µm voxel resolution were obtained of the proximal tibia. A hydroxyapatite phantom provided by the manufacturer was used as part of the weekly machine calibration process. The cemented implants were bisected in the sagittal plane using a 0.15 mm thick irrigated diamond blade (Isomet, Buehler Inc, Lake Bluff, IL). The lateral half was processed without decalcification and methyl methacrylate (MMA) embedded, sectioned in 0.15 mm intervals, and ground to 0.015 mm thickness for dynamic histomorphometry. The medial half was decalcified in 10% EDTA, paraffin embedded, and stained with hematoxylin and eosin, and tartrate-resistant acid phosphatase (TRAP) staining (Acid Phosphatase, Leukocyte Kit, Sigma-Aldrich, St. Louis, MO).
Xylenol orange tetrasodium salt
Manufactured by Merck Group
Sourced in United States
Xylenol Orange tetrasodium salt is a chemical compound used as a colorimetric indicator. It is a dark red-violet crystalline powder that changes color based on the pH of the solution it is added to.
Automatically generated - may contain errors
Lab products found in correlation
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Leukocyte acid phosphatase kit, by Merck Group (1 mentions)
Alizarin complexone, by Merck Group (1 mentions)
Calcein green, by Merck Group (1 mentions)
Calcein disodium salt, by Merck Group (1 mentions)
Arachidonic acid, by Merck Group (1 mentions)
Nordihydroguaiaretic acid, by Merck Group (1 mentions)
Rt pcr, by Thermo Fisher Scientific (1 mentions)
Accu check, by Roche (1 mentions)
4 ethylphenol, by Merck Group (1 mentions)
4 hydroxybenzyl alcohol, by Merck Group (1 mentions)
3 hydroxybenzyl alcohol, by Merck Group (1 mentions)
4 hydroxy 3 methoxybenzyl alcohol, by Merck Group (1 mentions)
4 isopropylphenol, by Merck Group (1 mentions)
2 hydroxybenzyl alcohol, by Merck Group (1 mentions)
Ferrous sulphate heptahydrate, by Merck Group (1 mentions)
Na2hpo4 7h2o, by Merck Group (1 mentions)
Nah2po4 2h2o, by Merck Group (1 mentions)
Cuso4 5h2o, by Merck Group (1 mentions)
D glucose, by Merck Group (1 mentions)
6 protocols using xylenol orange tetrasodium salt
1
Bone Mineralization Analysis via Fluorescence Imaging
2
Fluorochrome Labeling of Bone Formation
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Fluorochrome labels can be incorporated at sites of mineralization of bone and labels the front of mineralization at the time of administration [22 (link)]. By administering the labels at different times, bone formation can be followed with time. To observe bone apposition in the current study, rabbits were injected with two fluorochrome labels: xylenol orange (Xylenol Orange tetrasodium salt, 398187; Sigma-Aldrich, St Louis, MO, USA) and calcein green (Calcein disodium salt, 21030; Sigma-Aldrich). Two different administration schedules were used for analysis of early and late fluorochrome deposition. In each group, half of the animals were injected on Days 3 and 10 and the other half were injected on Days 7 and 21 with xylenol orange and calcein green respectively.
3
Fatty Acid and Oxidative Stress Biomarkers
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Arachidonic Acid (AA), nordihydroguaiaretic acid (NDGA) and Xylenol Orange tetrasodium salt were purchased from Sigma-Aldrich® (USA). Fish oil containing 35% eicosapentaenoic acid (EPA) and 40% docosahexaenoic acid (DHA) from Natufarma Labs (Buenos Aires, Argentina). Blood glucose and glycated haemoglobin (HbA1c) were determined by rapid test Accu-check by Roche (Switzerland). Total cholesterol (Chol), TG, C-Reactive Protein (CRP), enzymes: transaminases, pyruvic oxaloacetic, glutamic oxaloacetic and gamma glutamyltranspeptidase (GGTP) diagnostic kits were purchased from Wiener Lab® (Argentina). Rabbit polyclonal anti-synaptophysin (SYN) antibody was purchased from Dako Biotechnology (Denmark). IL-6 plasma levels were measured by immuno-ELISA OptEIA™ system and apoptosis determined by Flow Cytometry (Annexin V: PE) both detection kits were provided by BD Biosciences® (USA). Fatty acids methyl ester (FAME) were identified using a commercial standard Nu-check® (USA) [12 (link)]. Hippocampal IL-6 levels were determined by RT-PCR, supplies was purchased from Life Technologies (USA). Apoptotic cells in histological slides were identified by TunelS7100 ApopTag® Peroxidase in situ Apoptosis Detection Kit (Germany).
4
Enzymatic Synthesis of Phenolic Compounds
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4-Hydroxy-3,5-dimethoxybenzyl alcohol was from Alfa Aesar (Haverhill, MA, USA). 4-n-Butylphenol and 4-n-nonylphenol were from Lancaster Synthesis (Haverhill, MA, USA). Ferrous sulphate heptahydrate was from Merck (Burlington, MA, USA). 4-Allylphenol (chavicol) was from Quest International (Naarden, The Netherlands). 4-Allylphenol-2,6-dimethoxyphenol, 4-allyl-2-methoxyphenol (eugenol), 4-(aminomethyl)-2-methoxyphenol hydrochloride (vanillyl amine), 4-sec-butylphenol, 4-cyclohexylphenol, 4-cyclopentylphenol, 4-ethylphenol, 4-n-hexylphenol, 2-hydroxy-benzyl alcohol, 3-hydroxybenzyl alcohol, 4-hydroxybenzyl alcohol, 4-hydroxy-2-methoxybenzyl alcohol, 4-hydroxy-3-methoxybenzyl alcohol (vanillyl alcohol), 5-indanol, 4-isopropylphenol, 2-methoxy-4-methylphenol (para-creosol), 4-methylphenol (para-cresol), 4-n-pentylphenol, 4-n-propylphenol, 5,6,7,8-tetrahydronaphthol and xylenol orange tetrasodium salt were from Sigma-Aldrich (St. Louis, MO, USA). All other chemicals were from commercial sources and of the purest grade available. The pJ404-His-VAO and pBAD-EUGO-His plasmids were a kind gift from Prof. Dr. Marco Fraaije (University of Groningen).
5
Quantification of Copper-Containing Proteins
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d (+)-Glucose (≥99.5%), Na2HPO4·7H2O, NaH2PO4·2H2O, Xylenol Orange tetrasodium salt and CuSO4·5H2O were obtained from Sigma-Aldrich (St. Louis, MO, USA). Pierce™ BCA Protein Assay Kit was obtained from Thermo Scientific (Rockford, IL, USA).
Ultrapure water obtained from a Barnstead EasyPure water filter system (Thermo Scientific, Dubuque, IA, USA) was used for the preparation of all solutions. All solutions were filtered through a 0.2 μm sterile nitrocellulose filter.
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Ultrapure water obtained from a Barnstead EasyPure water filter system (Thermo Scientific, Dubuque, IA, USA) was used for the preparation of all solutions. All solutions were filtered through a 0.2 μm sterile nitrocellulose filter.
6
Lipid Peroxidation Assay Protocol
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Chloroform, methanol (HPLC grade), dichloromethane (HPLC grade), ethanol (Reagent grade), isopropanol and phenolphthalein were purchased from Fisher Scientific, Loughborough, UK. Xylenol orange tetrasodium salt (ACS reagent grade), cumene hydroperoxide (80%, technical grade), ammonium iron (II) sulphate hexahydrate (ACS reagent grade, 99%), pentanal, heptanal, octanal and nonanal and ethyl butyrate were supplied by Sigma–Aldrich-, Dorset, UK.
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