Ovaries were dissected as previously described but fixed for 20 minutes. Ovaries were denatured into 3 M HCl (sigma) diluted in 0.3% PBX for 15 minutes. HCl was removed, and ovaries were incubated 3 times for 5 minutes in 0.1 M Borax (Sigma) to neutralize HCl. After this step Borax solution was removed and ovaries were incubated in blocking solution with 0.5% NDS in PBX for 1 hour as previously described. Staining and mounting were performed as previously described.
Hydrochloric acid (hcl)
Manufactured by Merck Group
Sourced in Germany, United States, United Kingdom, India, Italy, France, Spain, Australia, China, Poland, Switzerland, Canada, Ireland, Japan, Singapore, Sao Tome and Principe, Malaysia, Brazil, Hungary, Chile, Belgium, Denmark, Macao, Mexico, Sweden, Indonesia, Romania, Czechia, Egypt, Austria, Portugal, Netherlands, Greece, Panama, Kenya, Finland, Israel, Hong Kong, New Zealand, Norway
Hydrochloric acid is a commonly used laboratory reagent. It is a clear, colorless, and highly corrosive liquid with a pungent odor. Hydrochloric acid is an aqueous solution of hydrogen chloride gas.
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Lab products found in correlation
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Sodium carbonate, by Merck Group (234 mentions)
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2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (179 mentions)
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Ascorbic acid, by Merck Group (173 mentions)
Quercetin, by Merck Group (145 mentions)
4 825 protocols using hydrochloric acid (hcl)
1
Ovary Tissue Fixation and Staining
2
Cobalt Extraction and Detection Protocol
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The salt of CoCl2.6H2O (Merck company, 98.0%) was acquired for the preparation of cobalt solution. Bis (2,4,4-trimethylpentyl monothiophosphinic acid) (Cyanex302) (Aldrich company, 90.0%), bis(2,4,4-trimethylpentyl)dithiophosphinic acid (Cyanex301) (Aldrich company, 90.0%), bis-(2,4,4-trimethylpentyl) phosphinic acid (Cyanex272) (Aldrich company, 90.0%), and Di-(2-ethylhexyl)phosphoric acid (D2EHPA) (Merck company, 95.0%), as the extractants and kerosene (99.9%) as the diluted phase from Merck company, were used in the experiments. Ethylene glycol from Sigma Aldrich, 99.5% was used as the non-aqueous system. Another diluted phase such as, chloroform (99.8%), 1-decanol (99.0%), and toluene (99.9%) was provided from Merck. HCl (37.0%, Merck) and NaOH (99.9%, Merck) solutions were used to reach the appropriate pH of solutions in the experiments. As reverse extraction agents, acids like sulfuric acid (96.0%, Merck), nitric acid (65.0%, Merck), and hydrochloric acid were utilized. Ascorbic acid (99.0%, Merck), ammonium thiocyanate (99.0%, Merck), acetone (99.0%, Merck), and hydrochloric acid were used as cobalt detectors using a UV spectrophotometer.
3
Analytical Characterization of Quinoa from Xinjiang
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Quinoa was from Xinjiang, China. All solvents and chemicals were of analytical reagent grade or higher. Equipment included a water bath, spectrophotometer, centrifuge, incubator, electrophoresis tank, microscope, swirl mixer, and 80 mesh sieve. The amylase activity was 3,700 U/g and the lipase activity was 100,000 U/g. The equipment and conditions used for high-performance liquid chromatography were as follows: Agilent 1100 HPLC system (Agilent, USA), including online degassing device (G1322A), quadruple pump (G1311A), autosampler (G1313A), VWD detector (G1314A), and 1525 Binary HPLC Pump (Waters Co.); methanol (chromatographic pure) and acetonitrile (chromatographic pure) were purchased from TEDIA (United States); tetrahydrofuran, triethylamine, hydrochloric acid, crystalline tetrahydrofuran, triethylamine, hydrochloric acid, and sodium acetate trichloroacetic acid were all of analytical grade in purity; water was Millipore ultrapure water. Seventeen kinds of amino acid standards (Aspartic acid, Histidine, Glutamic acid, serine, Glycine, Threonine, Alanine, Arginine, Tyrosine, Cystine, Valine, Methionine, PhenylAlanine, IsoLeucine, Leucine, Lysine, and Proline), OPA, FMOC, and angiotensin-converting enzyme (ACE) were purchased from Sigma (United States). FAPGG was purchased from Solarbio (China). S. aureus and E.coli was purchased from Bainer Biotech (China).
4
Carbon Steel Corrosion Inhibition Assessment
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In this investigation, the specimens employed were N80 carbon steel, having a composition of 0.31% carbon, 0.19% silicon, 0.92% manganese, 0.01% phosphorus, 0.008% sulfur, and 0.2% chromium, with the balance being iron, all percentages being by mass. The dimensions of these samples were meticulously cut to 1.5 cm in length, 1.2 cm in width, and 0.4 cm in thickness. The experimental setup involved immersing these samples in a corrosive medium, specifically a 15 wt.% HCl solution, obtained by diluting Sigma-Aldrich’s 37% HCl (Burlington, MA, USA) with distilled water. In preparation for the corrosion studies, the steel’s surface was meticulously refined using a series of abrasive papers, progressively moving from a grit of 600 to 1200. The study examined the effects of MeHDZ and HHDZ inhibitors at varying molar concentrations, precisely at 5 × 10−3 mol/L, 10−3 mol/L, and 5 × 10−4 mol/L, ensuring a comprehensive analysis of the corrosion behavior under these conditions.
5
Preparation of Aqueous Solutions
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Sodium sulfate (Na2SO4) was purchased from EMD Millipore Corporation (Germany). Hydrochloric acid (HCl) and sodium hydroxide (NaOH) were purchased from Sigma-Aldrich (USA). 50 mM Na2SO4, HCl with varied concentrations (0.1 mM, 1 mM, 10 mM, 100 mM and 1 M) and NaOH with varied concentrations (0.1 mM, 1 mM, 10 mM, 100 mM and 1 M) were prepared using ≥ 18 MΩ Milli-Q water produced from a Millipore system (Millipore Co., USA). PBS (Gibco™, 1×, pH 7.2) was purchased from Thermo Fisher Scientific (USA). Luria-Bertani (LB) Broth, Tryptic Soy Broth (TSB), Brain Heart Infusion (BHI) Broth and Nutrient Broth (NB) were purchased from Becton, Dickinson and Company (USA). Nuclease free water for PCR was purchased from Promega Corporation (USA).
6
Nitrate and Nitrite Quantification in STEMI and CTR Sera
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For measuring total nitrate and nitrite in the STEMI and CTR sera, the chemiluminescence detector Sievers Nitric Oxide Analyzer (NOA-280i; GE Water & Process Technologies, Analytic Instruments; Boulder, CO, USA) was used. Subsequent procedures were performed according to the operation and maintenance manual (Firmware Version 3.00 and later) provided by the manufacturer. The assay is based on the reduction of all nitrates and nitrites into nitric oxide (NO) by vanadium (III) chloride. NO reacts with ozone inside the NOA-280i to produce nitrogen dioxide (NO 2 ), which is sensitively detected by virtue of its chemiluminescence.
NO products (NOx) of the STEMI and CTR sera were analyzed by injecting 50 µL of each serum sample into a purge vessel containing a solution of vanadium (III) chloride (50 mmol/L; Sigma-Aldrich, Hamburg, Germany) in hydrochloric acid (HCl) (1 mol/L; Sigma-Aldrich, Hamburg, Germany) at 95 • C, continuously purged with a stream of nitrogen gas, connected to the NOA-280i. A gas bubbler between the purge vessel and the NOA-280i was filled with 15 mL of 1 M aqueous NaOH solution (Sigma-Aldrich, Hamburg, Germany) to prevent HCl vapors from entering the NOA-280i. Concentrations were calculated using the manufacturer's NO Analysis Software for Liquid (Version 3.21/Liquid, GE Water & Process Technologies, Analytic Instruments; Boulder, CO, USA).
NO products (NOx) of the STEMI and CTR sera were analyzed by injecting 50 µL of each serum sample into a purge vessel containing a solution of vanadium (III) chloride (50 mmol/L; Sigma-Aldrich, Hamburg, Germany) in hydrochloric acid (HCl) (1 mol/L; Sigma-Aldrich, Hamburg, Germany) at 95 • C, continuously purged with a stream of nitrogen gas, connected to the NOA-280i. A gas bubbler between the purge vessel and the NOA-280i was filled with 15 mL of 1 M aqueous NaOH solution (Sigma-Aldrich, Hamburg, Germany) to prevent HCl vapors from entering the NOA-280i. Concentrations were calculated using the manufacturer's NO Analysis Software for Liquid (Version 3.21/Liquid, GE Water & Process Technologies, Analytic Instruments; Boulder, CO, USA).
7
Collagen Quantification by Hydroxyproline
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Collagen was assessed by the measurement of hydroxyproline in tissue, following the methodology described by Reddy and Enwemeka (1996) [79 (link)], and Dwivedi et al. (2017) with modifications. Briefly, tissue samples frozen at −70 °C (Sample 2) were thawed and incubated for 15 h in an oven at 60 °C in uncapped microtubes. Samples were weighed and homogenized in POLYTRON® PT 3100 at 13,000 rpm with 6N hydrochloric acid (HCL) and transferred to capped glass test tubes. HCL ¨N was added in a proportion of 100 μL per 1 mg of dry tissue, homogenized and subjected to acid hydrolysis, thus incubating the samples in HCL (Sigma-Aldrich) at 130 °C for 4 h.
After incubation, the pH of the samples was adjusted to 7.0 with 2M sodium hydroxide (NaOH) (Sigma-Aldrich). Subsequently, 10 μL of each sample along with 90 μL of 0.056 M T-Chloramine solution were pipetted into 96-well plates incubated in the dark for 25 min. Afterwards, 100 μL of Ehrlich reagent [80 (link)] was added for the formation of the chromophore. Thus, the plates were incubated at 60 °C for 20 min as by performing the absorbance reading at 550 nm.
After incubation, the pH of the samples was adjusted to 7.0 with 2M sodium hydroxide (NaOH) (Sigma-Aldrich). Subsequently, 10 μL of each sample along with 90 μL of 0.056 M T-Chloramine solution were pipetted into 96-well plates incubated in the dark for 25 min. Afterwards, 100 μL of Ehrlich reagent [80 (link)] was added for the formation of the chromophore. Thus, the plates were incubated at 60 °C for 20 min as by performing the absorbance reading at 550 nm.
8
Silk Cocoon Dopamine Synthesis Protocol
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Dopamine hydrochloride (Sigma‐Aldrich, >99.9%), anhydrous ferric chloride FeCl3 (Sigma‐Aldrich, ≥99.99% trace metal basis), hydrochloric acid, HCl (Sigma‐Aldrich, reagent grade, ≥37%), and sodium carbonate (Na2CO3) were used without any further purification. Raw B. mori silk cocoons were purchased from Tagima Shogi (Japan).
9
Synthesis and Characterization of Hyperbranched Polyglycerol
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HPG (Mn = 949 g mol−1, Đ = 1.1, 14.6 OH per mol) was previously synthesized as described by Sunder et al. [20 (link)]. Oxalic acid dihydrate (OA, 99.5%, Synth, Diadema, Brazil), ibuprofen (IBU, DEG—lmportação de Produtos Químicos Ltda, São Paulo, Brazil) and phosphate buffer pH 7 (Tecnopon, Piracicaba, Brazil) were used as received. Hydrochloric acid (HCl, 37%, Sigma Aldrich, St. Louis, MO, USA) was used as received to prepare the HCl solutions. The water was distilled prior to use.
10
Standardization of Aqueous Solutions
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Zinc(II) solutions were prepared by dissolving the corresponding mass of ZnCl2 (99%, anhydrous, VWR) in water; the concentration was determined by complexometric titration against ethylenediaminetetraacetic acid (EDTA) standard solutions (Fisher Scientific). Standard hydrochloric acid (HCl) solutions were prepared from concentrated HCl (Sigma-Aldrich-Honeywell) and standardized with tris(hydroxymethyl)aminomethane (THAM) (Roche Diagnostics). CO2-free sodium hydroxide (NaOH) standard solutions were supplied by Fisher Scientific and were preserved from atmospheric CO2 by means of soda lime traps. Electrolyte solutions of sodium chloride (NaCl) were prepared from the pure salt (VWR). Citric acid monohydrate (VWR) and deferoxamine mesylate salt (Sigma-Aldrich) powders were used to prepare ligand solutions. Ultrapure water (R = 18 MΩ cm−1), grade A glassware, and analytical grade reagents were used throughout.
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