Freezing extender used in this study was Tris-glucose based extender containing 2.4% (w/v) Tris (BDH, England), 1.4% (w/v) citric acid (BDH, England), 0.8% (w/v) glucose (Merck, Germany), 0.06% (w/v) sodium benzyl penicillin (M&H manufacturing co., Ltd., Samutprakarn, Thailand), 0.1% (w/v) streptomycin sulphate (M&H manufacturing co., Ltd., Samutprakarn, Thailand), and different concentrations of L-carnitine (Sigma-Aldrich, St. Louis, MO, USA) (0 mM L-carnitine, 12.5 mM, 25 mM, and 50 mM). This extender was further supplemented with 3% and 7% (v/v) glycerol for extender 1 and 2, respectively. Thawing medium used in this study was prepared similarly to freezing extender except that it did not contain egg yolk, L-carnitine or glycerol (Axner et al., 2004 (link)).
Citric acid
Manufactured by BD
Sourced in United Kingdom, United States
Citric acid is a weak organic acid that occurs naturally in citrus fruits. It is commonly used in laboratory settings as a pH regulator, chelating agent, and preservative.
Automatically generated - may contain errors
Lab products found in correlation
L carnitine, by Merck Group (1 mentions)
Tetraethyl orthosilicate, by Merck Group (1 mentions)
D mannose, by Merck Group (1 mentions)
Potassium chloride (kcl), by Merck Group (1 mentions)
Lithium chloride (licl), by Merck Group (1 mentions)
Acrylamide, by Merck Group (1 mentions)
Calcium nitrate, by Merck Group (1 mentions)
Ammonium nitrate, by Merck Group (1 mentions)
Starch, by BD (1 mentions)
Annexin 5 fitc, by BD (1 mentions)
Falcon round bottom tubes, by BD (1 mentions)
Streptozotocin (stz), by Merck Group (1 mentions)
Sulphuric acid, by Avantor (1 mentions)
Glycerol, by Merck Group (1 mentions)
Na2so4, by Merck Group (1 mentions)
Fecl3, by Merck Group (1 mentions)
Cacl2.2h2o, by BD (1 mentions)
Nano3, by Merck Group (1 mentions)
Na2moo4 2h2o, by BD (1 mentions)
Tissue culture plate, by BD (1 mentions)
7 protocols using citric acid
1
L-Carnitine Supplemented Tris-Glucose Extender
2
Wheat Gluten Prolamine Extraction and Silica Coating
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For the prolamine extract, 20 g wheat gluten (>75% protein, Roth, Karlsruhe, Germany, 100 g) was extracted with 200 mL of 70 % ethanol/H2O dest solution. The solution was centrifuged for 10 min by 4000 rpm. Then, 10 mL of the supernatant was stirred with 0.5 g D-mannose (Sigma-Aldrich, St. Louis, MO, USA, 100 g). Tetraethyl orthosilicate (Sigma-Aldrich, 500 mL) was hydrolyzed beforehand with citric acid (VWR, BDH chemicals, Radnor, PA, USA, 1 kg), 3 mL was added to the solution. Then, 1.2 g of citric acid was added, and the solution was stirred for 24 h at 300 rpm. Trimethylethoxysilane (abcr, Karlsruhe, Germany, 1 L) was hydrolyzed and stirred for 1 h at 300 rpm. The paper was coated with 400 µm wet film of the barrier coating and dried at 80 °C in an oven for 30 min. The loaded particles were dispersed in the silica-sol based on trimethylethoxysilane, applied in wet film onto the barrier coating, and dried for 1 h at 80 °C.
3
Synthesis and Characterization of Organic-Inorganic Hybrid Materials
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Mercuric chloride (HgCl2), sodium hydroxide (NaOH), hydrazine (N2H2), NafionTM, ibuprofen, potassium nitrate (KNO3), sodium chloride (NaCl), calcium nitrate (Ca(NO3)2), ammonium nitrate (NH4NO3), acrylamide (electrophoresis grade) and lithium chloride (LiCl) (all analytical grade) were obtained from E. Merck; tetramethylammonium bromide (TMAB), tetramethylammonium iodide (TMAI) and tetrabutylammonium iodide (TBAI) from Fluka; nitric acid (HNO3), acetone, acrolein, acrylic acid, maleic acid, glucose, citric acid and starch from BDH, and potassium chloride from Sigma-Aldrich. Stock solution of 0.5% ibuprofen was prepared in methanol. 0.5% NafionTM solution was prepared in iso-propanol via intensive sonication. Stock solutions of 0.02 mol L−1 HgCl2, 0.4 mol L−1 NaOH, 0.05 mol L−1 LiCl, TMAB, TBAB, TMAI and TBAI and all other salts solutions were prepared in milli Q® deionized water (100 µS/cm).
4
Streptozotocin-Induced Diabetes Model
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Chemicals and drugs were sourced as follows: streptozotocin (Sigma-Aldrich Chemical Company, Missouri, St. Louis, USA); metformin and insulin (Sigma-Aldrich Chemical Company, St. Louis, USA); dimethyl sulfoxide (DMSO), citric acid, sodium citrate, FITC annexin-V, and BD Falcon round-bottom tubes (BD Biosciences, San Jose, CA); sulphuric acid (BDH Chemicals LTD, Poole, England); and ELISA kits (Biocom Africa, South Africa).
5
Semen Cryopreservation with LDL or Egg Yolk
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Freezing extender used in this study was prepared as followed: 2.4% (w/v) Tris (BDH, Leuven, Belgium), 1.4% citric acid (BDH, London, England), 0.8% glucose (Merck, Germany), 0.06% (w/v) sodium benzyl penicillin (M&H manufacturing co., Ltd., Samutprakarn, Thailand), 0.1% (w/v) streptomycin sulphate (M&H manufacturing co., Ltd., Thailand), and different concentrations of LDL (no LDL, 4%, 8%, 16%, 24% LDL) or 20% (v/v) egg yolk. These extenders were supplemented with 3% and 7% (v/v) glycerol (Sigma-Aldrich, St. Louis, MO, USA) for extender 1 and 2, respectively. Thawing medium used in this study was prepared according to Axner et al. (2004) (link).
6
Aminoglycoside Efficacy and Ion Effects
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The present study employed P. aeruginosa PA01 (ATCC 15,692) to evaluate the effect of aminoglycosides in conjunction with the presence/absence of Ca2+ and Mg2+ ions. The strain was cultivated on Evan’s media consisting of: NaH2PO4.2H2O (0.62 g/ l) (BDH, England), KCl (0.75 g/l) (Fluka), NaNO3 (0.849 g/l) (Sigma-Aldrich), Na2SO4 (0.28 g/l) (Sigma), citric acid (0.38 g/l) 0.02% (v/v) (BDH, England), glucose (1.8 g/l) (Thermo Fisher Scientific, UK), CaCl2.2H2O (0.037 g/l) (BDH, England), MgCl2 (0.11 g/l) (Amresco, United States American (USA)) and filter sterilised trace elements. The pH was adjusted to 7.2 prior to sterilisation by autoclaving. The concentrations of both Ca2+ and Mg2+ were amended to 1–5 mM when required.
Trace element solution contained ZnO (4.1 g/l), FeCl3 (3.2 g/l) (Sigma–Aldrich), MnCl2.4H2O (2.0 g/l), CuCl2 (1.4 g/l) (BDH, England), CoCl2.6H2O (4.8 g/l) (BDH, England), Na2MoO4.2H2O (0.0048 g/l) (BDH, England), NOB4O7.10 H2O (3.82 g/l) (Sigma, Japan) in 80 mL of concentrated HCl (Sigma–Aldrich, UK). Shaking flasks (250 ml) containing 50 mL of sterile Evan’s media and 50 μl of filtrated trace elements were inoculated with P. aeruginosa and incubated overnight at 30 °C, 250 rpm.
Trace element solution contained ZnO (4.1 g/l), FeCl3 (3.2 g/l) (Sigma–Aldrich), MnCl2.4H2O (2.0 g/l), CuCl2 (1.4 g/l) (BDH, England), CoCl2.6H2O (4.8 g/l) (BDH, England), Na2MoO4.2H2O (0.0048 g/l) (BDH, England), NOB4O7.10 H2O (3.82 g/l) (Sigma, Japan) in 80 mL of concentrated HCl (Sigma–Aldrich, UK). Shaking flasks (250 ml) containing 50 mL of sterile Evan’s media and 50 μl of filtrated trace elements were inoculated with P. aeruginosa and incubated overnight at 30 °C, 250 rpm.
7
Evaluation of Anthracycline Toxicity and Mitigation
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Clinical grade doxorubicin (DXH), epirubicin (EpiDXH) and daunorubicin(DNR) were obtained from Actavis, Italy, Pharmaceuticals and in view of the toxicity of DXH, EpiDXH and DNR, special care was paid. Ascorbic acid (AA) was purchased from Merck, Darmstadt. Disodium, while hydrogen phosphate, citric acid and EDTA of analytical purity grade were purchased from BDH. Tissue culture plates were purchased from Falcon, Oxnard, California, U.S.A. The reagents were applied without further purification.
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