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5 protocols using citronellal

1

Episodic Memory Assessment in Mice

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To assess effects of sex and mNMDAR function on episodic memory, mice were tested in tasks that used multiple odor cues and did not involve repetition or reward5 (link),21 (link),36 (link),37 (link). Mice were handled for 2-min the day before experimentation. Serial ‘What’ and ‘When’ tasks used plexiglass arenas (30×25-cm floor, 21.5-cm height) and two jars; the simultaneous ‘What’ and ‘Where’ tasks employed larger arenas (60×60-cm floor, 30-cm height) and 4 jars as described5 (link),21 (link). Each jar contained a filter paper scented with an odorant dissolved in mineral oil (~0.1 Pascals). The following odors, with letter identification, were used: (A) (+)-Limonene (≥97% purity, Sigma-Aldrich), (B) Cyclohexyl ethyl acetate (≥97%, International Flavors & Fragrances), (C) (+)-Citronellal (~96% Alfa Aesar), (D) Octyl Aldehyde (~99%, Acros Organics), (E) Anisole 99% (~99% Acros Organics). These odors were previously confirmed to be saliently balanced in both sexes21 (link).
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2

Antimicrobial Activity of Terpene Compounds

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Carveol and carvone were obtained from Sigma-Aldrich (Portugal) and citronellol and citronellal were obtained from Acros Organics (USA). Each compound (the compounds were weighed out leading to μg/mL units) was tested at various concentrations in the range of 0.066 to 3000 μg/mL in dimethyl sulfoxide (DMSO, 100%) (Sigma-Aldrich). The solutions of EOs components in DMSO did not exceed 10% (v/v) of the final volume of cell suspensions. Cell suspensions with DMSO (10%, v/v) and cell suspension without EOs components were used as controls. To assess the mode of action, each EOs component was tested at MIC concentration. All tests were performed in triplicate with three repeats.
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3

Olfactory Discrimination Training in Mice

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Studies used FVB129 mice as previously described72 (link). The mice were handled one day prior to the task. On training day, the mice were then placed into a plexiglas arena (60×60 cm floor, 30 cm walls) containing four empty glass cups (5.25 cm diameter x 5 cm height) with a metal lid with a single hole (~1. 5 cm diameter). The cups were removed from the chamber, and the animal rested undisturbed for 5 minutes. For the training session, the four cups were re-introduced with each containing one of the following odorants dissolved in mineral oil (final concentration of 0.1 Pascals): (A) (+)-Limonene (≥97% purity, Sigma-Aldrich), (B) Cyclohexyl ethyl acetate (≥97%, International Flavors & Fragrances Inc.), (C) (+)-Citronellal (~96% Alfa Aesar), and (D) Octyl Aldehyde (~99%, Acros Organics). The animal’s behavior was monitored over 5 minutes of odor exposure, and then they were returned to their home cage for 24 hours. For testing, the animals were allowed to explore the chamber with either odorant pairs (A:D) or (B:C) switched in position for 5 minutes.
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4

Methylene Blue Nanoparticle Synthesis

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Methylene blue (MB) dye 89%, Stearic acid 97%, oleic acid 90%, De-ionized water, polysorbate 20 (tween20), sodium glycocholate 97.5% and sodium tourocholate, 96% were purchased from Alfa Aesar (Thermo Fisher Scientific, Germany), while citronellal 93% and carvacrol 95% monoterpens were obtained from Acros Organics (Thermo Fisher Scientific, Germany). All the chemical reagents were used without any purification.
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5

Comprehensive Chemical Compound Acquisition

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Benzalkonium chloride (BKC), glycerol, parafluoro aniline (PFA), polyethylene glycol-400 (PEG-400), carvacrol, isopropyl alcohol, decanol, tri-isobutyl phosphate, terpinyl acetate, sodium fluorescein and agarose were purchased from Sigma-Aldrich (UK). Benzyl alcohol, citronellal, linalyl acetate, 1-bromohexane and methyl palmitate were purchased from Fischer Scientific, UK. Instant ocean salt was from Aquarium Systems (UK).
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