Aminoguanidine hemisulfate salt

Methylglyoxal (MG), propidium iodide, [3(4,5-dimethylthi-azol-2-yl)-2,5-diphenyl tetrazolium bromide] (MTT), neutral red, carbenoxolone disodium salt (CBX), aminoguanidine hemisulfate salt (AG), standard glutathione, o-phthaldialdehyde, metaphosphoric acid, L-glutamate, N-methyl-D-glucamine, 4-(2-hydroxyethyl) piperazine-L-ethanesulfonic acid (HEPES), and cell culture materials were purchased from Sigma (Saint Louis, MO, USA). Dulbecco's modified Eagle medium (DMEM) and Dulbecco's phosphate-buffered saline (DPBS) were purchased from Gibco BRL (Carlsbad, CA, USA). Fetal bovine serum was obtained from Cultilab (Campinas, SP, Brazil), and L-[2,3-³H] glutamate was purchased from Amersham International (United Kingdom). Polyclonal anti-EAAT1 (GLAST) and anti-EAAT2 (GLT-1) were purchased from Abcam (Cambridge, MA, USA), anti-EAAT3 was purchased from Novus Biologicals (Littleton, CO, USA), and polyclonal anti-glyoxalase 1 was purchased from Santa Cruz Biotechnology Inc. (Dallas, Texas, USA). All other chemicals were purchased from local commercial suppliers.

The catecholamine epinephrine hydrochloride (EPI), the NOS inhibitors S-methylisothiourea hemisulfate salt (SMIS) and aminoguanidine hemisulfate salt (AGH) as well as the NO donor sodium nitroprusside dihydrate (SNP) were purchased from Sigma-Aldrich. The irreversible inhibitor of soluble guanylyl cyclase 1H-[1,2,4]Oxadiazole[4,3-a]quinoxalin-1-one (ODQ) and the NO donor 3-morpholinosydnonimine chloride (SIN-1) were obtained from AdipoGen Life Sciences and the NOS inhibitors L-NG-nitroarginine methyl ester (L-NAME) hydrochloride, L-NG-nitroarginine (L-NNA) and 7-nitroindazole (7-NI) were purchased from Cayman chemicals. Additional chemicals were Levodopa (L-DOPA), obtained from Santa Cruz Biotechnology, as well as (+)-MK 801 maleate (MK-801) and Ifenprodil (+)-tartrate salt (ifenprodil), both obtained from Selleckchem. Stock solutions at 10^− 1 M for SMIS, AGH and SNP or at 10^− 2 M for remaining compounds were either prepared with autoclaved Milli-Q dH₂O or with DMSO (Sigma-Aldrich) for 7-NI and ODQ. Working solutions (10x concentrate of final concentration of treatment) for each compound were prepared prior to experiments with 1 μm filtered fresh seawater (FSW).

For in vivo competition experiments using EcN strains (Fig. 3D to F), mice were maintained on either filter-sterilized water (mock treatment) or a filter-sterilized solution of aminoguanidine hemisulfate salt (Sigma-Aldrich) (1 mg/ml) in water beginning on the same day as inoculation with EcN (3 days postinfection with T. gondii). In order to avoid any potential direct effect from aminoguanidine on gut microbiota structure, intraperitoneal injection (2 mg/mouse daily, dissolved in PBS) was used for 16S rRNA profiling experiments (Fig. 3A to C) from day 0 to the end of the experiment, and the control group included mice administered PBS (mock).