Methacrylated gellan gum (GG-MA) was obtained as previously described by Silva-Correia et al. [30 (link)]. Briefly, a solution of low-acyl gellan gum (Gelzan™ CM Gelrite®, Sigma-Aldrich) reacted with glycidyl methacrylate (GMA, 97%, Sigma-Aldrich) overnight at room temperature, with constant control of pH at 8.5 and under vigorous stirring. The reaction products were precipitated by the addition of cold acetone and further purified by dialysis (cellulose membrane, molecular weight cut-off 12 kDa, Sigma-Aldrich) against distilled water, for one week. Then, the obtained GG-MA was frozen at −80 °C, freeze-dried, and the final dry material was stored protected from light, in a dry place, until further use. GG-MA solutions of desired concentrations were prepared by dissolving the dry material in Milli-Q water using gentle agitation. Manganese (II) chloride powder (MnCl2 powder, Sigma-Aldrich) was used to prepare the MnCl2 aqueous solutions used as a supplement for GG-MA hydrogels. Artificial cerebrospinal fluid (aCSF) was prepared following the composition listed in Table 1 , and final pH adjustment to 7.3, with NaOH. For in vitro and in vivo assays, dry GG-MA was sterilized by UV light for 30 min in a laminar flow hood. All other materials and solutions were sterilized by filtration (0.22 µm filter).
Cellulose membrane
Manufactured by Merck Group
Sourced in United States, Germany, United Kingdom, Spain
The Cellulose Membrane is a semi-permeable membrane made from cellulose. It is used for the separation and purification of various substances in laboratory and industrial applications.
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Lab products found in correlation
Synergy ht, by Agilent Technologies (5 mentions)
Sodium deoxycholate, by Merck Group (4 mentions)
Nh4 2so4, by Merck Group (2 mentions)
Soya phosphatidylcholine, by Merck Group (2 mentions)
Pluronic f 127, by Merck Group (2 mentions)
Cholesterol, by Merck Group (2 mentions)
Ethanol, by Merck Group (2 mentions)
Sodium dodecyl sulfate (sds), by Merck Group (2 mentions)
Glycerol, by Thermo Fisher Scientific (2 mentions)
Uv 1601, by Shimadzu (2 mentions)
Pierce quantitative colorimetric peptide assay kit, by Thermo Fisher Scientific (2 mentions)
Gelzan cm gelrite, by Merck Group (1 mentions)
Vortex genie 2, by Scientific Industries (1 mentions)
Curcumin, by Merck Group (1 mentions)
Carrageenan, by Merck Group (1 mentions)
Cholesterol, by Kanto Chemical (1 mentions)
Tween 80, by Merck Group (1 mentions)
C18 column, by Waters Corporation (1 mentions)
Acquity uplc h class chromatography system, by Waters Corporation (1 mentions)
Rotary evaporator, by Heidolph (1 mentions)
70 protocols using cellulose membrane
1
Synthesis and Characterization of Methacrylated Gellan Gum Hydrogels
2
Preparation of Soluble Antigen Fraction from S. mansoni
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A soluble fraction in trichloroacetic acid (TCA-soluble fraction) was prepared from the crude extract of adult worms of S. mansoni, according to a previously described methodology [27 (link)], with modifications. An equal volume of 10% TCA was added to the total adult worm extract. After mixing vigorously for three cycles of 1 min, using a vortexer (Vortex Genie-2, Scientific Industries Inc., Bohemia, NY, USA), the suspension was subjected to centrifugation at 10,000 ×g, at 4°C for 45 min. The supernatant (containing the TCA-soluble fraction) was removed and then dialyzed against PBS on a cellulose membrane (Sigma-Aldrich) that retains substances with a molecular weight of 12,000 kDa or greater, with continuous stirring overnight at 4°C. We subsequently determined the protein content of the antigen solution (TCA-soluble fraction), which was then aliquoted and stored in a freezer at −80°C until further use. The TCA-soluble fraction was used for the sensitization of microplates to IgM (ELISA-IgM) antibodies [26 (link), 28 (link)].
3
Isolation and Purification of Parasite Eggs
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After perfusion of infected mice, livers were removed to recover parasite eggs. The eggs were homogenized and mechanically ground (Virtiz Precisa) for 40 min in 0.85% saline. The homogenate was centrifuged at 9,500 g for 1 h at 4°C. After 48 h of dialysis with a cellulose membrane (Sigma-Aldrich) against a 0.9% saline solution, an aliquot of the supernatant was measured for protein content (Nanodrop) to normalize protein concentration in the SEA-ELISA used to detect human anti-egg antibodies (see below).
4
Buffalo Cheese Whey Preparation
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The buffalo cheese whey was prepared by adding 0.6% (v/v) renin from A. niger and 0.5% (v/v) 0.5 mol. L-1 CaCl2 to buffalo milk at 35°C for 1 h. After casein precipitation, the whey was filtered in gauze [26 ] and dialyzed in cellulose membrane (12 kDa, Sigma) under constant magnetic stirring at 8°C. Periodic water exchange for lactose removal was also performed [27 ]. Fat was removed by adsorption in kaolin (20 g. L-1 w/v), with subsequent centrifugation at 7400 x g at 4°C for 30 min. The treated whey was stored at -18°C until further use.
5
Curcumin-loaded Lipid Nanoparticle Synthesis
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Sorbitan monooleate (Span 80, SP80), polyoxyethylenesorbitan trioleate (Tween 85, T85), cholesterol (Chol), curcumin, ethanol, methanol, and cellulose membrane with molecular weight cut-off = 14000 were purchased from Sigma-Aldrich (UK).
6
Electrochemical Expansion of Graphite
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Example 1
5 g of graphite flakes were compacted in a dialysis tubing of approximately 30 cm in length, flat with 25 mm, (Sigma-Aldrich, cellulose membrane) with a platinum wire placed inside. The portion of the dialysis tubing containing the graphite flakes was clamped on both sides with clips to maintain pressure on the graphite flakes. This assembly served as the cathode. The compact cathode was immersed in 3 L of a 0.1 M ammonium sulfate ((NH4)2SO4, from Sigma-Aldrich) aqueous electrolyte solution. A piece of graphite plate served as the anode. The cathode and anode were connected to a power supply. 10 V were applied to the dialysis bag with graphite flakes during electrochemical expansion, with a compliance current of ˜0.5 A. Electrochemical expansion time varied from 1-24 hours.
The electrochemically expanded graphite (EEG) was then washed via centrifugation with DI water five times. After washing, the EEG was transferred into a 400 mL Erlenmeyer flask, and the flask was filled up to 300 mL with DI water. After 30 seconds, the supernatant was transferred into a beaker for further characterization and treatments. The remaining sediment is unexpanded graphite.
7
Enzyme Purification from Broth Culture
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The broth cultures were centrifuged at 10,000× g for 20 min at 4 °C. The protein was then precipitated from the supernatant by stepwise ammonium sulfate addition, and the crude enzyme was collected at 60% saturation overnight at 4 °C. The solution was centrifuged at 10,000× g at 4 °C for 15 min. The precipitate was collected and dissolved in 20 mM Tris-HCl buffer. It was dialyzed by using dialysis tubing (cellulose membrane; 14 kDa molecular weight cut off, Sigma Aldrich, Sydney, Australia) in 1 L of the same buffer at 4 °C for desalting. The buffer was changed every two hours for 6 h. After dialysis, the supernatant was used as a partially purified enzyme for testing enzyme activity.
8
Characterization of Ginger Rhizome Extract
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Fresh ginger rhizome was purchased from a local market in Qena, Egypt. Identification of the plant was done at the Pharmacognosy Department, Faculty of Pharmacy, South Valley University, Qena, Egypt. A specimen (code Zo.82) was kept in the herbarium of the Pharmacognosy Department. Carrageenan, sodium deoxycholate, Tween 80, and cellulose membrane (molecular weight 12,000–14,000 Da cutoff) were obtained from Sigma Chemicals (St Louis, MO, USA). Phosphatidylcholine phospholipon 90G was purchased from Lipoid (Steinhausen, Switzerland). Cholesterol was obtained from Kanto Chemical (Tokyo, Japan). Sodium alginate was obtained from General Chemical and Pharmaceutical, Sudbury, UK. Chitosan (CS; low molecular weight, viscosity 100 cp, 98% degree of deacetylation) was obtained from Industrial Manufacturing, Tokyo, Japan. Hydroxypropyl methylcellulose 15,000 was obtained from El Gomhouria, Cairo, Egypt. Ethanol, potassium dihydrogen phosphate, and disodium hydrogen phosphate were purchased from the United Company for Chemicals and Medical Preparations (Cairo, Egypt). All other chemicals and solvents were of analytical grade and used as received.
9
Lignin-Oleate Membrane Formulation
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Four hundred milligram lignin and oleate HElignin were dissolved in 20 ml THF, respectively. The solutions were then transferred into dialysis tubing (cellulose membrane, Sigma-Aldrich) and placed into 1,500 ml distilled water for 7 days under slow stirring (Figure S1 ); the distilled water was changed every 24 h. The final concentration for the coating solution was 20 mg/ml (Zhao et al., 2016 (link)).
10
In vitro Release Test for Ciclopirox Olamine
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An in vitro release test was performed with flow-through cells (Sotax CE7 smart with pump). Cellulose membrane (Sigma Aldrich (St. Louis, USA)) was mounted in adapters for semi-solid formulations. The donor phase (1 ± 0.1 g) was placed into the adapter. The diffusion area was 1.33 cm2. The aqueous receptor medium was 50 mL of purified water. Selected conditions fulfilled the criteria of sink conditions. Samples of the solution were collected at 1, 2, 3, 4, 5, and 6 h and replaced with the same volume of fresh water.
All samples were analyzed by liquid chromatography using the Acquity UPLC H-Class chromatography system (Waters, Milford, MA, USA) equipped with DAD (Waters, Milford, MA, USA), performing detection at 303 nm. Validated UPLC method conditions were C18 column (130 Å, 1.7 µm, 2.1 mm × 50 mm, Waters, Milford, MA, USA), solvent A (acetonitrile) and solvent B 0.5% (v/v) of trifluoracetic acid in ultrapure water) ratio 40–60%; the injection volume was 5 μL; the flow rate was 0.7 mL/min; the column temperature was 25 °C. A standard calibration curve was built up by using standard solutions (4–324 µg/mL). All samples were filtered using a polyvinylidene difluoride filter (pore size 0.2 µm).
The cumulative amounts of released ciclopirox olamine expressed per unit area (cm2) were calculated and discussed in the results section.
All samples were analyzed by liquid chromatography using the Acquity UPLC H-Class chromatography system (Waters, Milford, MA, USA) equipped with DAD (Waters, Milford, MA, USA), performing detection at 303 nm. Validated UPLC method conditions were C18 column (130 Å, 1.7 µm, 2.1 mm × 50 mm, Waters, Milford, MA, USA), solvent A (acetonitrile) and solvent B 0.5% (v/v) of trifluoracetic acid in ultrapure water) ratio 40–60%; the injection volume was 5 μL; the flow rate was 0.7 mL/min; the column temperature was 25 °C. A standard calibration curve was built up by using standard solutions (4–324 µg/mL). All samples were filtered using a polyvinylidene difluoride filter (pore size 0.2 µm).
The cumulative amounts of released ciclopirox olamine expressed per unit area (cm2) were calculated and discussed in the results section.
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