Chemicals and reagents namely Dimethyl sulfoxide (DMSO), 2,2-diphenyl-1-picrylhydrazyl (DPPH), D-Pinitol, GTP, INH, Isopropyl β-D-1-thiogalactopyranoside (IPTG) and RIF were procured from Sigma (Aldrich, St. Louis, USA).
D pinitol
Manufactured by Merck Group
Sourced in United States
D-pinitol is a naturally occurring polyol compound that can be used as a laboratory reagent. It functions as a chemical standard and reference material for analytical and research applications.
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Lab products found in correlation
Myo inositol, by Merck Group (2 mentions)
D fructose, by Merck Group (2 mentions)
D glucose, by Merck Group (2 mentions)
Il 1β, by Fortis Life Sciences (2 mentions)
Il 6 enzyme linked immunosorbent assay elisa kit, by Fortis Life Sciences (2 mentions)
Aft6α, by Abcam (2 mentions)
Erk1 2, by Abcam (2 mentions)
Isopropyl β d 1 thiogalactopyranoside (iptg), by Merck Group (1 mentions)
Cesium triiodide, by Merck Group (1 mentions)
D galactose, by Merck Group (1 mentions)
Milli q rg apparatus, by Merck Group (1 mentions)
Methanol, by Merck Group (1 mentions)
Ammonium acetate, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
Trifluoroacetic acid, by Merck Group (1 mentions)
Chemstation, by Agilent Technologies (1 mentions)
Trypsin edta solution, by Solarbio (1 mentions)
Dulbecco s modified eagle s medium dmem, by Thermo Fisher Scientific (1 mentions)
Age bsa, by Abcam (1 mentions)
Ripk3, by Proteintech (1 mentions)
7 protocols using d pinitol
1
Biochemical Reagents Procurement
2
Quantification of Carbohydrate Standards
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D-pinitol (95% mass), L-chiro-inositol (95% mass), myo-inositol (≥98% mass), D-glucose, D-fructose, and D-galactose were purchased from Sigma-Aldrich (Steinheim, Germany). Water, acetonitrile, methanol (all solvents were of LC-MS grade purity), trifluoroacetic acid (≥99% purity), and ammonium acetate (≥99% purity) were purchased from Sigma-Aldrich (Steinheim, Germany). MALDI matrices such as 2,5-dihydroxybenzoic acid (DHB), α-cyano-4-hydroxycinnamic acid (HCCA) as well as cesium triiodide for mass calibration were purchased from Sigma-Aldrich (Steinheim, Germany). Water was obtained with a Milli-Q RG apparatus by Millipore (Millipore Intertech, Bedford, MA, USA).
3
Quantifying NSCs and Starch in Saplings
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The cambium and xylem tissues of 18 saplings (3 saplings×3 thermal conditions×2 irrigation regimes) were collected every 2 weeks (Supplementary Fig. S1 at JXB online) and NSCs were extracted following the procedure described in Giovannelli et al. (2011) . An Agilent 1200 series HPLC with a RID and a Shodex SC 1011 column and guard column, equipped with an Agilent Chemstation for the LC systems program, was used for assessment of soluble carbohydrates. A calibration curve was created for each carbohydrate using standard sucrose, raffinose, glucose, fructose (Canadian Life Science), and d -pinitol (Sigma-Aldrich). Total NSCs were calculated as the sum of soluble carbohydrate concentrations (raffinose, sucrose, glucose, pinitol, and fructose). Xylem powder was also used for starch extraction, performed according to Chow and Landhäusser (2004). The starch was solubilized with 0.1M NaOH and 0.1M acetic acid, and was hydrolysed enzymatically with an α-amylase solution at 2000U ml–1 and amyloglucosidase at 10U ml–1. PGO-colour reagent and 75% H2SO4 were added to the solution 24h later. Starch was assessed using a spectrophotometer at 533nm (Chow and Landhäusser, 2004 (link)).
4
Evaluation of D-pinitol's Effects on Diabetic Cells
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D-pinitol (purity: 95%, Lot No: 441252) and streptozotocin (STZ) were from Sigma Chemicals Co. (St. Louis, MO, USA). Paraformaldehyde (PFA), dimethyl sulfoxide (DMSO), bovine serum albumin (BSA), trypsin/EDTA solution, D-glucose, and 3-(4,5-dimethylthiazol)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) were purchased from Solarbio (Beijing, China). Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal bovine serum (FBS) were obtained from GIBCO (Grand Island, NE, USA). Hoechst 33258 and PI were purchased from Beyotime (Shanghai, China). AGE-BSA was purchased from Abcam (Cambridge, UK). ELISA kits and necrosulfonamide (NSA) were purchased from R&D systems (Minneapolis, MN, USA). Annexin V/PI Kit was from Key Gen (Nanjing, China). The primers were synthesized by Sangon Biotechnology (Shanghai, China). The MFG-E8 antibody was purchased from MBL (Woburn, MA, USA) and R&D (Minneapolis, MN, USA), and antibodies RIPK1, MLKL, phospho-MLKL, and β-action were purchased from Abcam (Cambridge, UK). RIPK3 and HMGB1 antibodies were purchased from Proteintech (Wuhan, China). All other reagents are standard commercial high purity materials.
5
Cytokine and Apoptosis Signaling Assay
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d-P initol (purity 95%) was purchased from Sigma-Aldrich Co. (St Louis,
MO, USA). Rats-specific TNF-α, IL-1β, and IL-6 enzyme-linked immunosorbent assay
(ELISA) Kit were obtained from Bethyl Laboratories Inc. (Montgomery, TX, USA).
The primary antibodies of caspase-3, caspase-9, caspase-12, AFT4 (Activating
transcription factor-4), AFT6α (Activating transcription factor-6α), XBP-1
(X-box binding protein 1), ERK-1/2 (Extracellular signal-regulated kinase-1/2),
and p38 were purchased from Abcam (Cambridge, MA, USA).
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MO, USA). Rats-specific TNF-α, IL-1β, and IL-6 enzyme-linked immunosorbent assay
(ELISA) Kit were obtained from Bethyl Laboratories Inc. (Montgomery, TX, USA).
The primary antibodies of caspase-3, caspase-9, caspase-12, AFT4 (Activating
transcription factor-4), AFT6α (Activating transcription factor-6α), XBP-1
(X-box binding protein 1), ERK-1/2 (Extracellular signal-regulated kinase-1/2),
and p38 were purchased from Abcam (Cambridge, MA, USA).
6
Comprehensive Honey Composition Analysis
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Most of the honey samples were collected from different regions of Poland (27 samples) from private beekeepers and manufacturers. Other samples were collected from Australia (3 samples), Romania (2 samples), Greece, Kameron, France, Czech Republic, Russia and Brazil (1 sample from each mentioned country). A full list of samples, including place of cultivation, year, manufacturer, and their physicochemical properties is presented in Table 1 .
Standard purity ≥95% D-pinitol, myo-inositol, D-chiro-inositol, ononitol, bornesitol, allo-inositol, cis-inositol, epi-inositol, D-glucose, D-fructose, D-maltose, lactose, D-sorbitol, and D-(+)-turanose and trimethylsilylimidazole (TMSI) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Standards of sucrose, xylose, quebrachitol, neo-inositol, muco-inositol with purity ≥98%, and pyridine were bought from Avantor (Gliwice, Poland). DPPH (2,2-Diphenyl-1-picrylhydrazyl), trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), and 70% EtOH, were purchased from Sigma Aldrich (St. Louis, MO, USA). Ultra-pure water was obtained from a Milli-Q water system (Millipore Bedford, MS, Boston, Massachusetts, USA).
Standard purity ≥95% D-pinitol, myo-inositol, D-chiro-inositol, ononitol, bornesitol, allo-inositol, cis-inositol, epi-inositol, D-glucose, D-fructose, D-maltose, lactose, D-sorbitol, and D-(+)-turanose and trimethylsilylimidazole (TMSI) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Standards of sucrose, xylose, quebrachitol, neo-inositol, muco-inositol with purity ≥98%, and pyridine were bought from Avantor (Gliwice, Poland). DPPH (2,2-Diphenyl-1-picrylhydrazyl), trolox (6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid), and 70% EtOH, were purchased from Sigma Aldrich (St. Louis, MO, USA). Ultra-pure water was obtained from a Milli-Q water system (Millipore Bedford, MS, Boston, Massachusetts, USA).
7
D-pinitol Suppresses Inflammatory Responses
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D-pinitol (purity 95%) was purchased from Sigma-Aldrich Co., St Louis, MO, USA. Rats-speci c TNF-α, IL-1β, and IL-6 enzyme-linked immunosorbent assay (ELISA) Kit were obtained from Bethyl Laboratories Inc., Montgomery, TX, USA. The primary antibodies of caspase-3, caspase-9, caspase-12, AFT4 (Activating transcription factor-4), AFT6α (Activating transcription factor-6α), XBP-1 (X-box binding protein 1), ERK-1/2 (Extracellular signal-regulated kinase-1/2), and P38 were purchased from Abcam, Cambridge, MA, USA.
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