Indomethacin (IND, γ polymorph), l -histidine (HIS), l -lysine (LYS), l -isoleucine (ISO), glycine (GLY); 99% purity, were purchased from Alfa Aesar (Heysham, England). l -arginine (ARG); ≥99.5% purity, was purchased from Sigma Aldrich (Nottingham, United Kingdom). All powders were used as received from the supplier, and properties of the raw chemicals are described in Table 1 . Ultrapure water was produced from a Pure Lab Ultra-System (London, United Kingdom), hydrochloric acid 32% and sodium hydroxide; ≥97% purity, were supplied by Sigma Aldrich (Poole, United Kingdom) and VWR international (Leicestershire, United Kingdom), respectively. A pH meter (OXOID limited, England) was calibrated with standard buffer solutions: phosphate buffers; pH 7 and 4 ± 0.2 at 25 ± 1 °C and used to determine the pH of the solutions. A temperature probe (YC-747UD data logger thermometer) was used to accurately determine the solution temperature (precision 0.1 °C).
L arginine arg
Manufactured by Merck Group
Sourced in United Kingdom, United States, Germany
L-arginine (ARG) is an amino acid that plays a crucial role in various biological processes. It is a natural compound found in the human body and serves as a precursor for the production of nitric oxide, which has important functions in the cardiovascular, immune, and nervous systems. ARG is commonly used in laboratory settings for research and scientific applications.
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Lab products found in correlation
L lysine, by Thermo Fisher Scientific (1 mentions)
Ph meter, by Thermo Fisher Scientific (1 mentions)
L histidine, by Thermo Fisher Scientific (1 mentions)
Peptide n glycosidase f pngase f, by New England Biolabs (1 mentions)
Neuraminidase, by New England Biolabs (1 mentions)
O glycosidase, by New England Biolabs (1 mentions)
Nonanal, by Merck Group (1 mentions)
L arginine, by Merck Group (1 mentions)
1 hexanol, by Merck Group (1 mentions)
3 protocols using l arginine arg
1
Solubility and pH Determination of Amino Acids
2
Glycation of HDL: Prevention and Deglycosylation
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We incubated HDL at the concentration of 1 mg/mL with glucose or methylglyoxal (MG; Sigma-Aldrich Co., St. Louis, MO, USA) at various concentrations (glucose: 100 and 500 mg/dL; MG: 0.06, 0.6, 1.6, 6, 16, 60 and 160 mM) at 37℃ for 24 or 96 hours. After treatment with MG, glycated HDL was dialyzed against sterile PBS at 4℃ for 48 hours. Naive HDL was prepared as the control in the same manner, except for the treatment with MG.
The enzymatic deglycosylation of HDL was performed using peptide-N-glycosidase F (PNGase F), neuraminidase, and o-glycosidase (New England Biolabs, Inc., Ipswich, MA), in accordance with the manufacturer’s protocol. To prevent glycation, we incubated HDL with aminoguanidine (AG; guanidine hydrochloride, Tokyo Kasei Co., Tokyo), L-arginine (Arg, Sigma-Aldrich Co., St. Louis, MO, USA), or alagebrium chloride (ALT-711, CHEMOS GmbH & Co., KG, Germany) prior to its treatment with MG.
The enzymatic deglycosylation of HDL was performed using peptide-N-glycosidase F (PNGase F), neuraminidase, and o-glycosidase (New England Biolabs, Inc., Ipswich, MA), in accordance with the manufacturer’s protocol. To prevent glycation, we incubated HDL with aminoguanidine (AG; guanidine hydrochloride, Tokyo Kasei Co., Tokyo), L-arginine (Arg, Sigma-Aldrich Co., St. Louis, MO, USA), or alagebrium chloride (ALT-711, CHEMOS GmbH & Co., KG, Germany) prior to its treatment with MG.
3
Honeybee Learning: Caffeine and Arginine
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Experiments were carried out at the Experimental Field of the University of Buenos Aires, Argentina (34°32′S, 58°26′W) between January and March 2019. Honeybees workers (Apis mellifera Linnaeus 1758) of unknown age were captured as they landed at the entrance of at least 10 different Langstroth hives of 10 frames each and composed of a mated queen, three or four frames of capped brood, food reserves and about 20,000 individuals.
The chemical compounds used to prepare the different treatments (see Table 1) were caffeine (Caff ) and L-arginine (Arg) (Sigma-Aldrich, Steinheim, Germany). To prepare the different solutions, we considered the caffeine and L-arginine concentrations previously used in honeybee learning studies (Chalisova et al., 2011; (link)Wright et al., 2013; (link)Lopatina et al., 2017) (link). In the case of caffeine, the concentrations used are within the range found in natural nectars (Kretschmar and Baumann, 1999; Wright et al., 2013) (link). In the case of L-arginine, the concentrations used are below the range found in natural nectars (Gardener and Gillman, 2001) (link).
For classical conditioning assays, we used 1-hexanol and nonanal (Sigma-Aldrich), both pure odours commonly present in floral fragrances (Knudsen et al., 1993) (link) that are known to be similarly learned by honeybees (Guerrieri et al., 2005) (link).
The chemical compounds used to prepare the different treatments (see Table 1) were caffeine (Caff ) and L-arginine (Arg) (Sigma-Aldrich, Steinheim, Germany). To prepare the different solutions, we considered the caffeine and L-arginine concentrations previously used in honeybee learning studies (Chalisova et al., 2011; (link)Wright et al., 2013; (link)Lopatina et al., 2017) (link). In the case of caffeine, the concentrations used are within the range found in natural nectars (Kretschmar and Baumann, 1999; Wright et al., 2013) (link). In the case of L-arginine, the concentrations used are below the range found in natural nectars (Gardener and Gillman, 2001) (link).
For classical conditioning assays, we used 1-hexanol and nonanal (Sigma-Aldrich), both pure odours commonly present in floral fragrances (Knudsen et al., 1993) (link) that are known to be similarly learned by honeybees (Guerrieri et al., 2005) (link).
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