Male C57BL/6 mice aged 8–10 weeks were obtained from the animal facility of Glenmark Pharmaceuticals Ltd. All animal experiments were approved by IAEC. Animals were maintained in an individually ventilated cages (IVC) in environmentally monitored air-conditioned room maintained at a temperature of 22 ± 3 °C, relative humidity of 40–70% and 12 h light/12 h dark cycle. Corn cob (BioCobb, AT&T) was used as the bedding material. Commercial pellet diet (Altromin, Germany) and community tap water passed through a reverse osmosis system (Millipore) were given. Water was provided ad libitum throughout the study period. Food was also provided ad libitum throughout the study period.
Reverse osmosis system
Manufactured by Merck Group
Sourced in Germany, Ireland
A reverse osmosis (RO) system is a type of water filtration technology that uses a semipermeable membrane to remove dissolved ions, molecules, and larger particles from water. The core function of an RO system is to purify water by forcing it through the membrane, which retains contaminants and allows clean water to pass through.
Automatically generated - may contain errors
Lab products found in correlation
Milli q unit, by Merck Group (2 mentions)
Penicillin g, by Merck Group (1 mentions)
Streptomycin sulfate, by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Concanavalin a (cona), by Merck Group (1 mentions)
β ionone, by Yuanye Bio-Technology (1 mentions)
Geraniol, by Yuanye Bio-Technology (1 mentions)
Methanol and acetonitrile hplc grade, by Merck Group (1 mentions)
Linalool, by Yuanye Bio-Technology (1 mentions)
Linalool oxide, by Yuanye Bio-Technology (1 mentions)
α ionone, by Yuanye Bio-Technology (1 mentions)
Sodium hydroxide (naoh), by Merck Group (1 mentions)
Adenine, by Merck Group (1 mentions)
Chromasolv plus, by Merck Group (1 mentions)
4 protocols using reverse osmosis system
1
Mice Housing and Husbandry Protocol
2
Fmoc-Amino Acid Incorporation into Phospholipid Monolayers
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Fmoc-phenylalanine (Fmoc-F) and Fmoc-tyrosine (Fmoc-Y) were purchased from Sigma-Aldrich. 1,2-Dimyristoyl-sn-glycero-3-phosphate (DMPA) phospholipid was purchased from Avanti Lipids and used as received. Solvents were used without further purification and purchased from Aldrich (Germany). Hydrochloric acid 37% was PRS-Codex purchased from Panreac and used as received. The initial 1 mM solutions of Fmoc-F and Fmoc-Y were prepared in dichloromethane. A mixture of chloroform : methanol (3 : 1 in volume) was used for dissolving DMPA at a concentration of 1 mM. The Fmoc-amino acids and the DMPA were co-spread in molar ratio DMPA : Fmoc amino acid 1 : 1. The choice of different solvents for the Fmoc-amino acid derivatives and the phospholipid was motivated to minimize any solubility problems. Considering the high miscibility of the solvents, no phase segregation was expected. Ultrapure water produced by a Millipore Milli-Q unit, and pre-treated by a Millipore reverse osmosis system (>18.2 MΩ cm) was used to prepare the acid solution for subphase. The subphase temperature was 21 °C with pH 2. All experiments were performed on tables with vibration isolation using the antivibration system MOD-2 S (Accurion, Göttingen, Germany) in a large class 100 clean room.
3
Phytochemical Analysis and Bioactivity Assay
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Acetonitrile and methanol (HPLC-grade) were purchased from Merck (Darmstadt, Germany). Standard products such as salidroside, acteoside, oleanolic acid, and ursolic acid were obtained from the National Institutes for Food and Drug Control (Beijing, China). L (-)-2-Octanol (as internal standard), linalool, linalool oxide, geraniol, α-ionone, and β-ionone were purchased from Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Light petroleum, boiling range of 80 to 120 °C, was purchased from J&K Chemical, Ltd. (Beijing, China). Water of HPLC grade was purified by a reverse osmosis system (Millipore, Ireland). All the reagents of HPLC grade were filtered across a 0.45 µm membrane before the experiment. Dimethyl sulfoxide (DMSO), 3-(4,5-dimethylthiazol-2-y 1)-2,5-diphenyltetrazolium bromide (MTT), trypan, concanavalin A (Con A), penicillin G, and streptomycin sulfate were obtained from Sigma Chemical Co. (St. Louis, MO, USA), and Gibco 1640 medium and fetal bovine serum (FBS) from Gibco Invitrogen Corp. (San Diego, CA, USA). Other reagents and chemicals were of analytical grade.
4
Lipid Monolayer Characterization Protocol
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The nucleolipid 1,2-dipalmitoyl-sn-glycero-3-(cytidine diphosphate) (ammonium salt) powder (≥ 99%), CDP-DG, and phospholipid 1,2-dimyristoyl-sn-glycero-3-phosphate (sodium salt) (≥ 99%), DMPA, were purchased from Avanti Polar Lipids and used as received. The purine bases, guanine hydrochloride (> 99%) and adenine (98%) were obtained from Merck and used as well.
The basic subphase at pH 11 was adjusted by NaOH (> 99%) and supplied from Merck. The solutions for CDP-DG and DMPA were prepared in chloroform and CHCl 3 /CH 3 OH 3/1, respectively, and used as spreading solvents. The pure solvents (Chromasol V® Plus) were purchased from Sigma Aldrich and used without further purification. Ultrapure water, produced by a Millipore Milli-Q unit, pre-treated by a Millipore reverse osmosis system (> 18.2 MΩ cm), was used as a subphase. The subphase temperature was kept at 21 °C.
The basic subphase at pH 11 was adjusted by NaOH (> 99%) and supplied from Merck. The solutions for CDP-DG and DMPA were prepared in chloroform and CHCl 3 /CH 3 OH 3/1, respectively, and used as spreading solvents. The pure solvents (Chromasol V® Plus) were purchased from Sigma Aldrich and used without further purification. Ultrapure water, produced by a Millipore Milli-Q unit, pre-treated by a Millipore reverse osmosis system (> 18.2 MΩ cm), was used as a subphase. The subphase temperature was kept at 21 °C.
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