Sodium chloride (nacl)

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Sodium chloride is a chemical compound with the formula NaCl. It is a white, crystalline solid that is commonly known as table salt. Sodium chloride is a vital mineral that plays a crucial role in maintaining the balance of fluids and electrolytes in the body.

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897 protocols using sodium chloride (nacl)

Porcine Heart Decellularization Protocol

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This study was authorized by the Animal Experimentation Ethics Committee (1166080618). Adult porcine hearts of both sexes were obtained under clean conditions from a slaughterhouse. Each heart weighing approximately 300 g, was submitted to decellularization protocol based on the previously literature [4 (link),8 (link)-10 (link),13 (link),14 (link)]. Protocols were tested, modified, and evaluated macroscopically and microscopically and the best one is described and detailed in this work.
For heart decellularization, connectors were purchased from peristaltic pump (Harvard Peristaltic Pump – Harvard Apparatus). About 4% sodium dodecyl sulfate (SDS) solution was prepared (Affymetrix, Santa Clara, CA). Hypertonic and hypotonic solutions were made from sodium chloride (Affymetrix). The phosphate-buffered saline (PBS) was prepared with sodium chloride (Affymetrix), sodium phosphate (Sigma-Aldrich, St. Louis, MO), potassium chloride (Sigma-Aldrich), and potassium phosphate (Sigma-Aldrich).

Delgado A.L., Carreira A.C., de Carvalho H.J., da Palma R.K., Sasahara T.H., de Carvalho C.M., León M., Barreto R.D, & Miglino M.A. (2021). Development of a new decellularization protocol for the whole porcine heart. Journal of Clinical and Translational Research, 7(4), 563-574.

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PMMA and Polystyrene Microparticle Preparation

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For experiments involving non-fluorescent polymethyl methacrylate (PMMA) microparticles, a saline buffer was first prepared by mixing 2 g of NaCl (Thermo Fisher Scientific Inc., Waltham, MA, USA) with 10 mL of DI water. The PMMA microparticles (Cospheric LLC, Santa Barbara, CA, USA) with continuous size range 2–32 µm in diameter were then mixed with the prepared saline buffer at a concentration of ~3 million particles/mL (0.134 g in 50 mL of saline buffer). Buffer solution was used in preparing sample solution in order to match the particle density of 1.2 g/cm³ to achieve neutral buoyancy and minimize particle sedimentation in the syringe during experiments.
For experiments involving fluorescent polystyrene microparticles, a saline buffer was first prepared by mixing 1.6 g of NaCl (Fisher Scientific Inc., Waltham, MA, USA) with 10 mL of DI water to match the particle’s density of 1.06 g/cm³. The polystyrene particles of diameter 7.32 µm and 15.45 µm (Bangs Laboratories Inc., Fishers, IN, USA), 18.67 µm and 26.3 µm (Polysciences Inc., Warrington, PA, USA) were mixed with the prepared saline buffer at a concentration of ~5 million particles/mL. Tween 80 was added at 0.1% v/v (Fisher Scientific, Waltham, MA, USA) to all particle solutions to minimize aggregation and avoid clogging.

Bogseth A., Zhou J, & Papautsky I. (2020). Evaluation of Performance and Tunability of a Co-Flow Inertial Microfluidic Device. Micromachines, 11(3), 287.

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Extraction and Characterization of Lemon Peel Pectin

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Lemon peel was received in dried and milled state from a local supplier. Extraction of the alcohol insoluble residue (AIR) from lemon peel was performed using ethanol (C 2 H 5 OH, technical, Chem-lab, Zedelgem, Belgium) and acetone (C 3 H 6 O, technical, VWR, Pennsylvania, USA). Acid pectin extraction on the AIR was performed using 7N nitric acid (HNO 3 , 65%, Merck, New Jersey, USA). Furthermore, standardized tap water containing 0.2% sodium chloride (NaCl, analytical grade, Fisher scientific, New Hampshire, USA) and 0.015% dehydrated calcium chloride (CaCl 2 •H 2 O, 99.5%, Chem-lab, Zedelgem, Belgium) in ultrapure water (organic free, 18 MΩ cm resistance), supplied by a Simplicity™ water purification system (Millipore, Billerica, USA), was used to make AR suspensions.
Either sodium carbonate (NaCO 3 , 99.8%, VWR, Pennsylvania, USA) or hydrochloric acid (HCl, Fisher scientific, New Hampshire, USA) was used for pH adjustments whereas sodium chloride (NaCl, analytical grade, Fisher scientific, New Hampshire, USA) or calcium chloride (CaCl 2 •H 2 O, 99.5%, Chem-lab, Zedelgem, Belgium) was used to increase the salt content. Acridine orange (Sigma-Aldrich, Missouri, USA) was used to stain cell wall polymers for epifluorescence microscopy.

Willemsen K.L.D.D., Panozzo A., Moelants K., Cardinaels R., Wallecan J., Moldenaers P, & Hendrickx M. (2018). Effect of pH and salts on microstructure and viscoelastic properties of lemon peel acid insoluble fiber suspensions upon high pressure homogenization. Food Hydrocolloids., 82.

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Single-Nucleus ATAC-Seq Library Preparation

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Cells were collected in 1.5 mL centrifuge tube and added lysis buffer which consisted of 10 mM Tris-HCl pH7.5 (Thermo Fisher Scientific, USA), 10 mM NaCl (Thermo Fisher Scientific, USA), 3 mM MgCl₂ (Thermo Fisher Scientific, USA), 0.1% Tween-20 (Sigma, USA), 0.1% NP-40 (Roche, Switzerland), 0.01% Digitonin (Sigma, USA), and 1% bovine serum albumin (BSA, BBI, UK). Then the lysates were centrifuged at 500 g for 5 min at 4°C and the supernatant was discarded. The obtained nuclei were then washed three times with ATAC wash buffer (10 mM Tris-HCl pH7.5 (Thermo Fisher Scientific, USA), 10 mM NaCl (Thermo Fisher Scientific, USA), 3 mM MgCl₂ (Thermo Fisher Scientific, USA), 0.1% Tween-20 (Sigma, USA), 1% bovine serum albumin (BSA, BBI, UK)). Nuclei were then transposed using transposase (BGI, China) and were resuspended in nuclear resuspension buffer. Transposed single-nucleus suspensions were converted to barcoded scATAC-seq libraries, through procedures including droplet encapsulation, pre-amplification, emulsion breakage, capture beads collection, DNA amplification and purification. The libraries were sequenced on the ultra-high-throughput DIPSEQ T1 sequencer with PE 50-bp read length.

Wang S., Xie J., Zou X., Pan T., Yu Q., Zhuang Z., Zhong Y., Zhao X., Wang Z., Li R., Lei Y., Yin J., Yuan Y., Wei X., Liu L., Liu S., Yang H, & Wu L. (2022). Single-cell multiomics reveals heterogeneous cell states linked to metastatic potential in liver cancer cell lines. iScience, 25(3), 103857.

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Preparation and Analysis of Cellular Lysates

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Total cell lysates were prepared in a buffer containing 50 mM Tris-HCl (pH 7.8; Thermo Fisher Scientific), 137 mM NaCl (Thermo Fisher Scientific), 10 mM NaF (Thermo Fisher Scientific), 1 mM EDTA (Thermo Fisher Scientific), 1% Triton X-100 (Thermo Fisher Scientific), 10% glycerol (Thermo Fisher Scientific), and the protease inhibitor cocktail (Roche) through 3 freeze/thaw cycles. Tissue lysates were prepared by homogenizing in a buffer containing 50 mM Tris (pH 7.6; Thermo Fisher Scientific), 130 mM NaCl (Thermo Fisher Scientific), 5 mM NaF (Thermo Fisher Scientific), 25 mM β-glycerophosphoate (Thermo Fisher Scientific), 1 mM sodium orthovanadate (Thermo Fisher Scientific), 10% glycerol (Thermo Fisher Scientific), 1% Triton X-100 (Thermo Fisher Scientific), 1 mM dithiothreitol (Thermo Fisher Scientific), 1 mM phenylmethanesulfonyl fluoride (PMSF) (Thermo Fisher Scientific), and the protease inhibitor cocktail (Roche). After centrifugation (12,000g, 4°C for 10 minutes), tissue lysates were separated on SDS-polyacrylamide gel (SDS-PAGE) and analyzed using the following antibodies: rabbit anti-UCP1 (UCP11-A, Alpha Diagnostic), rabbit anti-HSP90 (sc-7947, Santa Cruz Biotechnology Inc.), rabbit anti–phospho-PKA substrate (9624, Cell Signaling Technology), rabbit anti-TH (ab112, Abcam), and mouse anti-tubulin (sc-32293, Santa Cruz Biotechnology Inc.).

Chen Z., Zhang P., Liu T., Qiu X., Li S, & Lin J.D. (2024). Neuregulin 4 mediates the metabolic benefits of mild cold exposure by promoting beige fat thermogenesis. JCI Insight, 9(1), e172957.

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Cultivation of M. tuberculosis in 7H9 medium

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M. tuberculosis was grown at 37°C in 7H9 Middlebrook medium (Becton Dickinson Biosciences) with 0.2% (vol/vol) glycerol (Sigma-Aldrich) and 0.05% (vol/vol) Tween 80 (Sigma-Aldrich), supplemented with either 10% albumin-dextrose-NaCl (ADN; 5% [wt/vol] bovine serum albumin fraction V [BSA; Roche], 2% [wt/vol] glucose [Sigma-Aldrich], 0.85% [vol/vol] NaCl [Thermo Fisher Scientific]) or 10% oleic acid-albumin-dextrose-catalase (OADC; 5% [wt/vol] bovine serum albumin fraction V [BSA; Roche], 2% [wt/vol] glucose [Sigma-Aldrich], 0.85% [vol/vol] NaCl [Thermo Fisher Scientific], 0.056% [vol/vol] oleic acid [Sigma-Aldrich], 4% [wt/vol] catalase [Sigma-Aldrich]). M. tuberculosis H37Rv was grown in 7H9 medium supplemented with 10% ADN, while clinical isolates were grown with 10% OADC supplement.

Rollo R.F., Mori G., Hill T.A., Hillemann D., Niemann S., Homolka S., Fairlie D.P, & Blumenthal A. (2023). Wollamide Cyclic Hexapeptides Synergize with Established and New Tuberculosis Antibiotics in Targeting Mycobacterium tuberculosis. Microbiology Spectrum, 11(4), e00465-23.

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Preparation of Buffers and Cell Culture Media

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Buffer solutions were prepared in nanopure water as follows. 10x PBS contained 1370 mM NaCl, 27 mM KCl, 80 mM Na₂HPO₄, and 20 mM KH₂PO₄ (Thermo Fisher Scientific, Waltham, MA). 10x Citrate buffer contained 100 mM sodium citrate dihydrate (Sigma-Aldrich, St. Louis, MO), 1370 mM NaCl, and 27 mM KCl (Thermo). Dulbecco’s Modified Eagle Medium (DMEM) supplement was added at 13.4 g/L and Rosewell Park Memorial Institute (RPMI) supplement was added at 10.4 g/L, according to manufacturer’s instructions. 2 mM triethanolamine (TEOA) was prepared in 1x PBS at pH 7.4. The pH was measured using an Orion Star A111 pH meter (Thermo) and adjusted with 1 M NaOH or HCl after the addition of all chemicals and/or supplements.

Jansen L.E., Negrón-Piñeiro L.J., Galarza S, & Peyton S.R. (2018). Control of Thiol-Maleimide Reaction Kinetics in PEG Hydrogel Networks. Acta biomaterialia, 70, 120-128.

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Vibrio Isolation and Preservation Protocol

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Tryptic soy agar (TSA, Oxoid) was used as the nonselective media, and thiosulfate citrate bile salts sucrose (TCBS, Oxoid) for the isolation of Vibrio strains. They were both prepared according to the manufacturer’s instructions, with the addition of 2% NaCl (Oxoid). Additionally, tryptic soy broth (TSB, Oxoid) with 2% NaCl was used for the cryopreservation of the strains at −80 °C in a final dilution of 15% glycerol.

Lattos A., Bitchava K., Giantsis I.A., Theodorou J.A., Batargias C, & Michaelidis B. (2021). The Implication of Vibrio Bacteria in the Winter Mortalities of the Critically Endangered Pinna nobilis. Microorganisms, 9(5), 922.

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High Concentration Saline Gradients for Power

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Aqueous sodium chloride solutions were prepared using 99% NaCl (Alfa-Aesar, Lancashire, UK) and deionised water. A 0.51 M concentrated feed and 0.02 M dilute feed, corresponding to standard sea/river water equivalent concentrations, were prepared, in order to benchmark data form the high concentration salinity gradient. A 4 M concentrated feed and 0.02 M dilute feed were used to develop the high concentration gradient, which has been previously identified as the most suitable concentration gradient to promote high power density [23 (link)]. The electrode rinse solution contained 0.1 M K₃Fe(CN)₆, 0.1 M K₄Fe(CN)₆ (Fisher Scientific, Leicestershire, UK) and 2 M NaCl (Alfa-Aesar, Lancashire, UK).

Hulme A.M., Davey C.J., Tyrrel S., Pidou M, & McAdam E.J. (2021). Scale-up of reverse electrodialysis for energy generation from high concentration salinity gradients. Journal of Membrane Science, 627, 119245.

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Sodium Chloride Solutions for Electrolyte Membranes

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Sodium chloride (NaCl) solutions were prepared for the concentrated and dilute feeds using 99% NaCl (Alfa-Aesar, Lancashire, UK) and deionised water. For the concentrated feed, solutions of 0.5 M, typically used to represent sea water in the literature, and 4 M were prepared. For the dilute feed, a 0.02 M solution was prepared. The electrode rinse consisted of 0.1 M potassium ferricyanide (K₃Fe(CN)₆), 0.1 M potassium ferrocyanide (K₄Fe(CN)₆)(Fisher Scientific, Leicestershire, UK) and 2 M or 0.25 M NaCl (Alfa-Aesar, Lancashire, UK) depending on the concentration of the feed to limit water transport to the electrolyte and was continuously recirculated to the stacks. For experiments using the large commercially available stacks, a feed reservoir containing 5 L of electrode rinse solution was prepared and 1 L for the 10 cm × 10 cm stack and wrapped in tin foil to avoid exposure to light.

Hulme A.M., Davey C.J., Tyrrel S., Pidou M, & McAdam E.J. (2021). Transitioning from electrodialysis to reverse electrodialysis stack design for energy generation from high concentration salinity gradients. Energy Conversion and Management, 244, None.

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