GSH concentrations in the lens were determined according to the method described by Sedlak and Lindsay, with minor modifications [19 (link)]. Briefly, lenses were homogenized in 500 μl of 0.1 M sodium phosphate buffer (pH 8.0) and centrifuged at 20,000 ×g for 20 min at 4 °C. The supernatant was mixed with an equal volume of 20% trichloroacetic acid, incubated for 15 min at 4 °C, and then centrifuged at 20,000 ×g for 30 min at 4 °C. Supernatant was diluted with 50 μl sodium phosphate buffer per 1 mg wet lens weight, and then 500 μl was added to 50 μg of dithionitrobenzene (DTNB; Wako Pure Chemical Industries Ltd.) and incubated for 30 min at room temperature. Absorbance at 412 nm was then measured in a microplate reader infinite M200 (Tecan Ltd., Männedorf, Switzerland). The GSH concentration in the lens was determined with a standard curve prepared using GSH (Wako Pure Chemical Industries Ltd.).
Glutathione (gsh)
Manufactured by Fujifilm
Sourced in Japan
The GSH is a piece of laboratory equipment manufactured by Fujifilm. It is designed to perform specific functions in the laboratory setting. The core function of the GSH is to provide a solution for the task at hand, without any further interpretation or extrapolation.
Automatically generated - may contain errors
Lab products found in correlation
L cysteine, by Nacalai Tesque (2 mentions)
Tapi 2, by Peptide Institute (2 mentions)
Bafilomycin a1, by Cayman Chemical (2 mentions)
Na2s2, by Dojindo Laboratories (2 mentions)
Na2s3, by Dojindo Laboratories (2 mentions)
Methanol, by Fujifilm (2 mentions)
Milli q system, by Merck Group (2 mentions)
Tempol, by Merck Group (2 mentions)
Sod from human erythrocytes, by Merck Group (2 mentions)
Sod assay kit wst, by Dojindo Laboratories (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Isoflurane, by Fujifilm (2 mentions)
Sprague dawley rats, by Japan SLC (2 mentions)
Metaphosphoric acid, by Fujifilm (2 mentions)
Sodium selenite, by Fujifilm (2 mentions)
Caffeine, by Fujifilm (2 mentions)
Xanthine, by Fujifilm (1 mentions)
Inosine, by Fujifilm (1 mentions)
Uridine, by Fujifilm (1 mentions)
Cytidine, by Fujifilm (1 mentions)
26 protocols using glutathione (gsh)
1
Quantification of Lens Glutathione Levels
2
Authentic Compound Characterization
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Authentic compounds were purchased from Wako Chemical Co. (Tokyo, Japan), including glutathione, hypoxanthine, xanthine, inosine, uridine, cytidine, guanine, guanosine, and L-ascorbic acid. All solvents were LC–MS grade (Kanto Chemical, Tokyo, Japan).
3
Synthesis and Characterization of Eudesmane Lactones
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Alantolactone (catalog number: S8318; Selleck Chemicals, Houston, TX, USA), bafilomycin A1 (item number: 119038; Cayman Chemical, Ann Arbor, MI, USA), costunolide (product number: 032-13731; Wako Pure Chemical Industries, Osaka, Japan), glutathione (product number: 073-02013; FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), L-cysteine (product number: 10309-41: NACalai Tesque), Z-Leu-Leu-Leu-H (aldehyde) (also known as MG-132) (code number: 3175-v; Peptide Institute, Osaka, Japan), NAC (product number: 00512-84; NACalai Tesque), parthenolide (catalog number: P0667-5MG; Sigma, St. Louis, MO, USA), and TAPI-2 (code number: INH-3852-PI; Peptide Institute) were purchased as commercial products. (11S)-2α-bromo-3-oxoeudesmano-12,6α-lactone (also known as santonin-related compound 2: SRC2) (1 ), (11S)-3-oxoeudesmano-12,6α-lactone (2 ), 2α-bromo-3-oxoeudesm-11(13)-eno-12,6α-lactone (3 ), 3-oxoeudesm-11(13)-eno-12,6α-lactone (4 ), (11S)-3-oxoeudesm-1-eno-12,6α-lactone (5 ), (11S)-3β-hydroxyeudesm-1-eno-12,6α-lactone (6 ), 3-oxoeudesma-1,11(13)-dieno-12,6α-lactone (also known as (+)-tuberiferin) (7 ), and 3β-hydroxyeudesma-1,11(13)-dieno-12,6α-lactone (8 ) were synthesized as previously described [36 (link),69 (link),70 (link)].
4
Sulfur Compound Preparation and Characterization
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All methods were performed in accordance with the guidelines and regulations of chemical substance management and approved by the committees of chemical substance management in the Sanyo-Onoda City University, Musashino University and National Institute of Neuroscience, National Center of Neurology and Psychiatry. Na2S2 (Dojindo, Kumamoto, Japan), Na2S3 (Dojindo), Na2S (Wako pure chemicals, Osaka, Japan), l -cysteine (Wako), and glutathione (Wako) were dissolved at 0.1 M in ultrapure water. These stock solutions were stored at − 80 °C, and were used within a week. 5,7-Dichlorokynurenic acid (DCKA) was obtained from Tocris Bioscience (Avonmouth, UK). d -Serine and the other chemicals were from Wako Pure Chemical Industries, Ltd. (Osaka, Japan).
5
Preparation and Use of Sulfur Compounds
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All methods were performed in accordance with the guidelines and regulations of chemical substance management and approved by the committees of chemical substance management in the National Institute of Neuroscience, National Center of Neurology and Psychiatry. Diethylamine NONOate (DEA/NO) (Cayman Chemical, Ann Arbor, MI) and Angeli’s salt (Cayman Chemical) were dissolved at 0.1 M in 10 mM NaOH. Na2S2 (Dojindo, Kumamoto, Japan), Na2S3 (Dojindo), Na2S (Wako pure chemicals, Osaka, Japan), L-cysteine (Cys) (Wako), glutathione (Wako), dithiothreitol (Wako) were dissolved at 0.1 M in ultrapure water. These stock solutions were stored at −80 °C, and were used within a week. NaCN (Wako) was dissolved at 0.5 M in a 0.1 M HEPES buffer (pH 7.4) and stored at −20 °C.
6
Cucurbitacin B Inhibition Study
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Cucurbitacin B (Tokyo Chemical Industries Co., Ltd., Tokyo, Japan), bafilomycin A1 (Cayman Chemical Company, Ann Arbor, MI, USA), MG-132 (Peptide Institute, Osaka, Japan), TAPI-2 (Peptide Institute), N-acetyl-L-cysteine (Nacalai Tesque), glutathione (FUJIFILM Wako Pure Chemical Corporation, Osaka, Japan), L-cysteine (Nacalai Tesque), ascorbic acid (Nacalai Tesque), Trolox (FUJIFILM Wako Pure Chemical Corporation), cytochalasin D (Cayman Chemical Company), and jasplakinolide (Nacalai Tesque) were purchased for the present study. Human recombinant TNF-α was provided by Dainippon Pharmaceutical Co., Ltd. (Osaka, Japan).
7
MeHg Exposure in Mice
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Two-month-old male mice were randomly divided into two groups: a vehicle group and an MeHg-exposed group. MeHg (Tokyo Chemical Industry, Tokyo, Japan) was dissolved at 30 ppm in distilled water containing equimolar amounts of glutathione (Fujifilm Wako Pure Chemical, Osaka, Japan) and administered via drinking water for 8 weeks as previously described (Fujimura et al. 2009 (link)). The vehicle group received drinking water containing only glutathione.
8
Bacterial Culturing for Oral Pathogens
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All chemicals were used as received without further purification. Glutathione (reduced form, 98% purity), methotrexate (MTX, 98.0% purity), methanol (99.7% purity), D2O (99.9% purity), and silver nitrate (99.9% purity) were purchased from FUJIFILM Wako Pure Chemical Corporation Ltd. (Osaka, Japan). Sodium borohydride (NaBH4, 99.99% purity) was purchased from Sigma-Aldrich (St. Louis, MO, USA).
Bacterial strains used in this study were S. mutans ATCC 35668, A. actinomycetemcomitans ATCC 29522, and P. gingivalis ATCC 33277. These strains were kept frozen until analysis. Bacterial stocks were anaerobically and statically incubated in brain heart infusion (BHI) broth (Pearlcore®, Eiken Chemical, Co., Ltd., Tokyo, Japan) supplemented with 0.1% antibiotics (gramicidin D and bacitracin, FUJIFILM Wako Pure Chemical Corporation Ltd.) and 1% sucrose (FUJIFILM Wako Pure Chemical Corporation Ltd.) for S. mutans; 1% yeast extract (FUJIFILM Wako Pure Chemical Corporation Ltd.) for A. actinomycetemcomitans; and 0.5% yeast extract, 0.0005% hemin, and 0.0001% menadione for P. gingivalis. Anaerobic incubation was carried out in an AnaeroPack system using anaerobic jars (Mitsubishi Gas Chemical Company, Inc., Tokyo, Japan).
Bacterial strains used in this study were S. mutans ATCC 35668, A. actinomycetemcomitans ATCC 29522, and P. gingivalis ATCC 33277. These strains were kept frozen until analysis. Bacterial stocks were anaerobically and statically incubated in brain heart infusion (BHI) broth (Pearlcore®, Eiken Chemical, Co., Ltd., Tokyo, Japan) supplemented with 0.1% antibiotics (gramicidin D and bacitracin, FUJIFILM Wako Pure Chemical Corporation Ltd.) and 1% sucrose (FUJIFILM Wako Pure Chemical Corporation Ltd.) for S. mutans; 1% yeast extract (FUJIFILM Wako Pure Chemical Corporation Ltd.) for A. actinomycetemcomitans; and 0.5% yeast extract, 0.0005% hemin, and 0.0001% menadione for P. gingivalis. Anaerobic incubation was carried out in an AnaeroPack system using anaerobic jars (Mitsubishi Gas Chemical Company, Inc., Tokyo, Japan).
9
Acetaminophen Metabolism and Glutathione
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APAP was purchased from Sigma-Aldrich (St. Louis, MO, USA). OPZ was purchased from LKT Laboratories (St. Paul, MN, USA). Glutathione (GSH) was purchased from Wako Pure Chemical Industries (Osaka, Japan). All the other chemicals and solvents used in this study were of biological or analytical grade.
10
Allantopyrone A Synthesis Protocol
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Allantopyrone A was prepared as described previously. 13 Glutathione (reduced form) was commercially obtained from Wako Pure Chemical Industries (Osaka, Japan).
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