The commercial standards: rutin, β-carotene, β-amyrin, β-sitosterol, lupeol, ursolic acid and linoleic acid were procured (Sigma Aldrich, USA). The AR grade organic solvents (hexane, ethyl acetate, chloroform and methanol) and p-anisaldehyde were purchased (BDH, UK). HPLC grade methanol (Merck, Germany) was used for the preparation of standard stock solutions (1000 μg/ml) and their serial dilutions (β-amyrin and β-sitosterol: 10–100 μg/ml; lupeol and ursolic acid: 10–120 μg/mL).
β sitosterol
Manufactured by Merck Group
Sourced in United States, Germany, China, Australia, Italy, France, Japan
β-sitosterol is a phytosterol compound commonly found in plants. It is a naturally occurring sterol that is structurally similar to cholesterol. β-sitosterol is often used as a reference standard in analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Campesterol, by Merck Group (50 mentions)
Stigmasterol, by Merck Group (49 mentions)
Cholesterol, by Merck Group (19 mentions)
Methanol, by Merck Group (14 mentions)
5α cholestane, by Merck Group (13 mentions)
Lupeol, by Merck Group (12 mentions)
Ergosterol, by Merck Group (11 mentions)
Squalene, by Merck Group (11 mentions)
Brassicasterol, by Merck Group (11 mentions)
β amyrin, by Merck Group (10 mentions)
Ursolic acid, by Merck Group (10 mentions)
Quercetin, by Merck Group (9 mentions)
Gallic acid, by Merck Group (9 mentions)
Rutin, by Merck Group (8 mentions)
Fucosterol, by Merck Group (7 mentions)
α tocopherol, by Merck Group (7 mentions)
Desmosterol, by Merck Group (7 mentions)
Acetonitrile, by Merck Group (7 mentions)
Linoleic acid, by Merck Group (7 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (7 mentions)
146 protocols using β sitosterol
1
Quantification of Phytochemical Standards
2
Yeast Fermentation under Anaerobic Conditions
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Triplicate fermentations were carried out in 330-mL glass flasks equipped with a sampling port containing 250 mL of SM medium for each strain. These flasks were also equipped with water-filled fermentation locks in order to maintain self-generated anaerobiosis during fermentation. Steampasteurisation of the flasks with SM was carried out at 100°C for 15 min to ensure asepsis without destroying thermolabile compounds such as vitamins. Then, the medium was saturated with sterile air for 20 min while being constantly agitated to balance the levels of dissolved and headspace oxygen. Furthermore, after sparging and before inoculation, under sterile conditions the SM was supplemented with 5 mg/L phytosterols in order to meet the lipid requirements of the yeast during anaerobic growth. The phytosterol stock solution, prepared with a commercial solution of βsitosterol >70% (Sigma 85541) that also contains other sterols, was composed of 20 g/L β-sitosterol (Sigma-Aldrich, St. Louis, MO, USA), Tween 80 (Sigma-Aldrich)/absolute ethanol (1:1, v/v) in the stock solution. The medium was fermented under isothermal conditions at 22°C with continuous agitation at 250 rpm.
3
Extraction and Characterization of Phytosterols
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n-Propanol (for synthesis), squalene (99%) and a technical β-sitosterol product (55.5% β-sitosterol, 34.2% campesterol [16 (link)]) were purchased from Merck (Darmstadt, Germany). Silica gel 60 (for column chromatography), 3-phenyl propionic acid (99%), 5α-cholestane (97%), cholesterol (> 99%) and stearic acid (99%) were ordered from Sigma-Aldrich (Steinheim, Germany). Dichloromethane (99.8%), mEthanol (HPLC gradient grade) and n-hexane (HPLC gradient grade) were purchased from Th. Geyer (Renningen, Germany). Ethanol (distilled before use, 7.4% water content), sodium chloride (99%), pyridine (99%) and sulphuric acid (96%) were obtained from Carl Roth (Karlsruhe, Germany). Tripalmitin (95%, containing traces of dipalmitoylglycerol), cholesteryl stearate (97%) and elaidic acid (99%) were from Fluka (Taufkirchen, Germany). Palmitic acid (98%) was purchased from Riedel de Häen (Seelze, Germany). 8-Phenyl octanoic acid (97%) and n-hexanol (99%) were from Thermo Fisher Scientific (Kandel, Germany). The trimethylsilylating reagent SILYL-991 was ordered from Macherey–Nagel (Düren, Germany). Nitrogen and helium 5.0 quality originated from Westfalen company (Münster, Germany). Dihydrolanosterol [19 ] and α-tocopherol [23 (link)] were isolated autonomously with CCC. Demineralised water was obtained through the in-house supply (Hohenheim, Germany).
4
Phytosterol Effects on Cell Lines
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Immortalized human embryonic kidney cells (HEK293.T), human microglia (CHME3; a kind gift from prof. dr. M. Tardieu, Universite Paris-Sud, France66 (link)), human oligodendrocytes (MO3.13), mouse neuroblastoma expressing APPswe (N2a/APPswe; a kind gift from prof. dr. T.W. Kim, Colombia University, USA67 (link)), and monkey kidney cells (COS7) were used for in vitro experiments. All cell lines were cultured in DMEM (Sigma-Aldrich) containing 10% heat-inactivated FCS (Invitrogen, Merelbeke, Belgium) and 100 U penicillin/100 µg streptomycin/ml (Invitrogen), at 37 °C/5% CO2. For phytosterol treatment, cells were incubated for 18 hours in culture medium without FCS containing the Eastern plants extracts, brassicasterol (Sigma-Aldrich), β-sitosterol, (Sigma-Aldrich), fucosterol (Sigma-Aldrich), stigmasterol (analytic confirmed purity of 99,9%), phytosterol mix (containing 60% β-sitosterol, 25% campesterol, and 15% stigmasterol; kindly provided by Ingmar Wester Raisio, Finland), T0901317 (Cayman Chemicals, Huissen, the Netherlands), ethanol (VWR), or DMSO (Sigma-Aldrich).
5
Cloning and Purification of Recombinant Proteins
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All restriction enzymes and DNA modifying enzymes were obtained from New England Biolabs (MA, USA). Plasmid pET-30a was obtained from Novagen (Germany). Escherichia coli DH10β and BL21 (DE3) were used as the host for plasmid cloning and protein expression, respectively. Nickel-nitrilotriacetic acid-agarose was purchased from Qiagen (USA). DNA and protein markers were acquired from New England Biolabs, β- NADPH, β-sitosterol and stigmasterol were acquired from Sigma-Aldrich (USA). Other materials used in this study were of analytical grade and were commercially available.
6
Analysis of Phytochemical Compounds from Various Sources
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The following substances and solvents were used in the study: procyanidins B1, B2, C1, A1, A2, A4, (+)-catechin, (–)-epicatechin, quercetin, quercitrin (quercetin-3-O-rhamnoside), isoquercitrin (quercetin-3-O-glucoside), avicularin (quercetin-3-O-arabinofuranoside), guaiaverin (quercetin-3-O-arabinopyranoside), rutin (quercetin-3-O-rutinose), reynoutrin (quercetin-3-O-xyloside), kaempferol, nicotiflorin (kaempferol-3-O-rutinoside), afzelin (kaempferol-3-O-rhamnoside), astragalin (kaempferol-3-O-glucoside), chlorogenic acid (3-O-caffeoylquinic acid), neochlorogenic acid (5-O-caffeoylquinic acid), cryptochlorogenic acid (4-O-caffeoylquinic acid), p-coumaric acid, arbutin, hydroquinone, α-amyrin, β-amyrin, β-sitosterol, lupeol, erythrodiol, maslinic acid, oleanolic acid, acetonitrile, acetone, and methanol, which were purchased from Sigma–Aldrich (Steinheim, Germany); hyperoside (quercetin-3-O-galactoside), procyanidin B3, uvaol, friedelin, 6″-O-acetylisoquercitrin (quercetin-3-O-(6″-acetylglucoside)), betulin, and betulinic and corosolic acids from Extrasynthese (Genay, France); ursolic acid from Carl Roth (Karlsruhe, Germany); and trifluoroacetic acid from Merck (Darmstadt, Germany). All used chemicals were of HPLC grade. Ultrapure water used in this study was prepared with Milli–Q® 180 (Millipore, Bedford, MA, USA) water purification system.
7
Fatty Acid Analysis in Cell Lines
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The reference standards of the fatty acids β-sitosterol, δ-tocopherol, lipopolysaccharide (LPS), CDCl3, Griess reagent (N5751) and N-nitro-l -arginine-methyl ester (l -NAME) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Methanol, tetrahydrofuran acetonitrile, sodium hydroxide, and boron trifluoride/Methanol were obtained from Fisher Scientific (Pittsburgh, PA, USA). DMEM (Dulbecco’s modified Eagle medium), FBS (fetal bovine serum), trypsin-EDTA, l -glutamine, penicillin/streptomycin were obtained from HyClone (South Logan, UT, USA). The tetrazolium salt (MTT) kit was purchased from Roche Applied Science (Mannheim, Germany). Isoflurane was purchased from DS Pharma Animal Health Co. (Osaka, Japan).
8
Antioxidant and Cytotoxic Evaluation
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Folin-Ciocalteu reagent, gallic acid, quercetin, ascorbic acid, curcumin, β-sitosterol, lupeol, 1,1-diphenyl-2-picrylhydrazyl (DPPH), 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt (ABTS), 3-(4, 5-dimethylthiazolyl-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), sulforhodamine B (SRB), Hoechst 33258 dye, crystal violet, propidium iodide were purchased from Sigma Chemicals Co. (St. Louis, MO, USA). All other chemicals and solvents were of analytical grade and purchased from the usual sources.
9
Analysis of Corn Silk Phytosterols
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CS samples of two cultivars, corn silk (zhengdan 958, xianyu 335), were harvested in September 2013 in Kaifeng farmland of China, pulverized and sifted through a 80-mesh sieve to obtain the powdered samples. The CS powder was dried at 80 °C overnight and stored with dark bags in dry environment prior to the experiments.
Stigmasterol (assay purity 97%), campesterol (assay purity 98%), β-sitosterol (assay purity 97%) were all acquired from Sigma Aldrich (Sydney, Australia). All other chemicals and reagents were purchased locally and of HPLC or analytical grade. Ultrapure water was used throughout the experiments and obtained using a Millipore water purification system (Element A10).
Stigmasterol (assay purity 97%), campesterol (assay purity 98%), β-sitosterol (assay purity 97%) were all acquired from Sigma Aldrich (Sydney, Australia). All other chemicals and reagents were purchased locally and of HPLC or analytical grade. Ultrapure water was used throughout the experiments and obtained using a Millipore water purification system (Element A10).
10
Characterization of Drug-Resistant Tuberculosis
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Mtb H37Rv was initially isolated from a patient with pulmonary tuberculosis in 1905 by Albert Calmette and Camille Guérin at the Pasteur Institute in Paris, France. The strain was maintained and distributed by American Type Culture Collection (ATCC 27294) centre. For our study, a clinical sample from extensively drug-resistant Mycobacterium tuberculosis h37rv was obtained from the district Tuberculosis laboratory (Guna, Gujarat, India). It has a rpoB gene mutation (gene accession number: JN037845) and has resisted the first, second, and third lines of TB drugs in primary screening. In addition, we also obtained a sample for drug-sensitive tuberculosis from the same laboratory. The culture was then inoculated into the Middlebrook medium (Himedia) supplied with 0.05% Tween 80 and albumin-dextrose-catalase. The culture was maintained by transferring in the fresh medium every 30 days. Madasiatic acid, Asiaticoside A(ASA), β-sitosterol, Cystargamide B, Myristic acid, Palmitic acid, Kalmeghin, Oleanolic acid, Longicyclene, Oleanane, and Amyrin were purchased from Sigma-Aldrich.
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