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4 protocols using sodium saccharine

1

Metformin Chewing Gum Development

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Metformin powder and the chewing gum base were kindly provided by Minoo Industrial Company (Tehran. I.R Iran). Glycerin, aspartame, sodium hydroxide, potassium hydrogen phosphate, sodium saccharine, potassium acesulfame, chloroform, and dichloro-methane (Merck Chemical Company, Germany) were purchased from market. Spearmint essence was bought from Parea-Quinica Company (Spain).
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2

Artemether-based Topical Formulation

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Artemether was a gift from Ipca Laboratories Ltd. India. Other ingredients include k-carrageenan and absolute ethanol, (Sigma Aldrich, Germany), propylene glycol, sodium saccharine andpeppermint oil (Merck, Germany). All other reagents and solvents were analytical grade and were used as procured.
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3

Alcohol, Sweeteners Consumption Study

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EtOH, Sucrose and sodium saccharin were used as drinking solutions. EtOH (99.8%) solutions (PanReac AppliChem, Barcelona, Spain) diluted to 5, 10, 20, 30 or 40% (v/v) were prepared in sterilized tap water. Sucrose solutions were prepared by dissolving ≥ 99.5% Sucrose (Sigma-Aldrich Química S.A. Madrid, Spain) in sterilized tap water to 5, 10, 20 or 40% (w/v). Sodium saccharine (Sigma-Aldrich, ≥ 98%) solutions at 0.13, 0.26, 0.53 or 1.06% (w/v) were dissolved in sterilized tap water. Concentrations of all drinking solutions were based on previous studies (Rhodes et al., 2005 (link), 2007 (link); Kamdar et al., 2007 ; Pastor et al., 2011 (link); Tarragón et al., 2012 (link)) as well as on our own pilot studies. In the case of EtOH, a direct study of the influence of availability of different concentrations and number of tubes was included in the present study (Experiment 1). NTX hydrochloride (Sigma-Aldrich) was diluted in 0.9% physiological saline and administered intraperitoneally (IP; 10 mL/kg injection volume). The dose range of NTX (4, 8 or 16 mg/kg) used in the present study was based on previous work (Kamdar et al., 2007 ; Tarragón et al., 2012 (link); Fultz et al., 2017 (link)).
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4

Saccharin Preference Test for Rats

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The SP task [23 (link)] was conducted by placing two 400 mL bottles in the rat’s home cage. One bottle contained tap water, the other contained a 0.05% sodium saccharine (Sigma-Aldrich) solution. Bottles were weighed before placement and after 72 hours exposure. Bottle position was alternated every 24 hours to reduce side bias. Saccharine preference was calculated as a percent of total intake. Trials occurred weekly after injury, and a pre-injury baseline assessment was taken. Sessions were conducted each week and initiated immediately after PR sessions on Friday and concluded immediately before FST on Monday.
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