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Characterisation and comparison of temporal release profiles of nitric oxide generating donors.

Bradley SA, Steinert JR - J. Neurosci. Methods (2015)

Bottom Line: We found that donors such as NOC-5 and PAPA-NONOate decayed substantially within days, whereas SNP and GSNO showed greater stability releasing consistent levels of NO over days.In all donors tested, the amount of released NO differs between frozen and unfrozen stocks.Fluorescent and amperometric approaches to measure NO concentrations yield a wide range of levels.

View Article: PubMed Central - PubMed

Affiliation: MRC Toxicology Unit, Hodgkin Building, University of Leicester, Leicester LE1 9HN, UK.

No MeSH data available.


Related in: MedlinePlus

Temporal release of NO from GSNO. (A) Average profiles of NO release over 30 min from fresh GSNO stock at 100, 200 and 300 μM. (B) Average release profile of NO yield from two day old stock at 100, 200 and 300 μM over 30 min. (C) Average NO release profiles from three day old GSNO at 100, 200 and 300 μM over a 30 min recording period. (D) Average NO release profiles of 4 week old frozen stock at 100, 200 and 300 μM over a 30 min recording period, note the saturation of the signal at 300 μM. (E) Box and whisker plots displaying the plateau concentration range of NO recorded using GSNO stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the curve using GSNO stock over 3 consecutive days and frozen stock, note that values for 300 μM frozen stock are a slight underestimation due to saturation of the signal.
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fig0025: Temporal release of NO from GSNO. (A) Average profiles of NO release over 30 min from fresh GSNO stock at 100, 200 and 300 μM. (B) Average release profile of NO yield from two day old stock at 100, 200 and 300 μM over 30 min. (C) Average NO release profiles from three day old GSNO at 100, 200 and 300 μM over a 30 min recording period. (D) Average NO release profiles of 4 week old frozen stock at 100, 200 and 300 μM over a 30 min recording period, note the saturation of the signal at 300 μM. (E) Box and whisker plots displaying the plateau concentration range of NO recorded using GSNO stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the curve using GSNO stock over 3 consecutive days and frozen stock, note that values for 300 μM frozen stock are a slight underestimation due to saturation of the signal.

Mentions: GSNO is a S-nitrosothiol which is synthesised through the S-nitrosation of primary and secondary thiols (Feelisch, 1998). S-nitrosothiols are catalysed by thiols, light and transition metals, and additionally, their stability in solution depends on temperature, pH and oxygen pressure. Here we sought to characterise the NO release from fresh GSNO stock over a three day period and stock frozen for four weeks. On day one NO release resulted in a steady increase to plateau at ∼24 min at 100 μM, ∼17 min at 200 μM and ∼16 min at 300 μM (Fig. 5A). NO release on day one to three was similar with plateaus reached by ∼24, ∼19 and ∼20 min for 100, 200 and 300 μM, respectively (Fig. 5A–C). Stock frozen for a month produced different NO release curves compared to the fresh stock. There is a rapid release of NO which increases with increased concentration; the response then decays over the 30 min recording period without reaching a plateau (Fig. 5D). Plateau concentrations of NO over the three consecutive days from fresh stock are remarkably stable with very little decay. The NO concentration from 100 μM GSNO is 145 ± 21 nM on day one, 145 ± 13 nM on day two and 136 ± 18 nM on day three. 200 μM GSNO liberates 283 ± 29 nM, 279 ± 26 nM and 262 ± 28 nM NO for day one, two and three, respectively. Finally, 300 μM GSNO resulted in NO plateau concentrations of 381 ± 32 nM, 384 ± 39 nM and 359 ± 38 nM for day one, two and three, respectively. Frozen stocks have lower plateau values of NO release (100 μM: 105 ± 6 nM, 200 μM: 193 ± 6 nM, 300 μM: 279 ± 13 nM) (Fig. 5E, Table 1). Total amount of NO released during the recording period is very stable across the subsequent days, however, the frozen stock showed a bell shape NO release curve. This resulted in a larger total amount of NO released (AUC) in comparison to fresh stock (Fig. 5F).


Characterisation and comparison of temporal release profiles of nitric oxide generating donors.

Bradley SA, Steinert JR - J. Neurosci. Methods (2015)

Temporal release of NO from GSNO. (A) Average profiles of NO release over 30 min from fresh GSNO stock at 100, 200 and 300 μM. (B) Average release profile of NO yield from two day old stock at 100, 200 and 300 μM over 30 min. (C) Average NO release profiles from three day old GSNO at 100, 200 and 300 μM over a 30 min recording period. (D) Average NO release profiles of 4 week old frozen stock at 100, 200 and 300 μM over a 30 min recording period, note the saturation of the signal at 300 μM. (E) Box and whisker plots displaying the plateau concentration range of NO recorded using GSNO stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the curve using GSNO stock over 3 consecutive days and frozen stock, note that values for 300 μM frozen stock are a slight underestimation due to saturation of the signal.
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fig0025: Temporal release of NO from GSNO. (A) Average profiles of NO release over 30 min from fresh GSNO stock at 100, 200 and 300 μM. (B) Average release profile of NO yield from two day old stock at 100, 200 and 300 μM over 30 min. (C) Average NO release profiles from three day old GSNO at 100, 200 and 300 μM over a 30 min recording period. (D) Average NO release profiles of 4 week old frozen stock at 100, 200 and 300 μM over a 30 min recording period, note the saturation of the signal at 300 μM. (E) Box and whisker plots displaying the plateau concentration range of NO recorded using GSNO stock on day one, two and three and frozen stock. (F) Box and whisker plots depicting the area under the curve using GSNO stock over 3 consecutive days and frozen stock, note that values for 300 μM frozen stock are a slight underestimation due to saturation of the signal.
Mentions: GSNO is a S-nitrosothiol which is synthesised through the S-nitrosation of primary and secondary thiols (Feelisch, 1998). S-nitrosothiols are catalysed by thiols, light and transition metals, and additionally, their stability in solution depends on temperature, pH and oxygen pressure. Here we sought to characterise the NO release from fresh GSNO stock over a three day period and stock frozen for four weeks. On day one NO release resulted in a steady increase to plateau at ∼24 min at 100 μM, ∼17 min at 200 μM and ∼16 min at 300 μM (Fig. 5A). NO release on day one to three was similar with plateaus reached by ∼24, ∼19 and ∼20 min for 100, 200 and 300 μM, respectively (Fig. 5A–C). Stock frozen for a month produced different NO release curves compared to the fresh stock. There is a rapid release of NO which increases with increased concentration; the response then decays over the 30 min recording period without reaching a plateau (Fig. 5D). Plateau concentrations of NO over the three consecutive days from fresh stock are remarkably stable with very little decay. The NO concentration from 100 μM GSNO is 145 ± 21 nM on day one, 145 ± 13 nM on day two and 136 ± 18 nM on day three. 200 μM GSNO liberates 283 ± 29 nM, 279 ± 26 nM and 262 ± 28 nM NO for day one, two and three, respectively. Finally, 300 μM GSNO resulted in NO plateau concentrations of 381 ± 32 nM, 384 ± 39 nM and 359 ± 38 nM for day one, two and three, respectively. Frozen stocks have lower plateau values of NO release (100 μM: 105 ± 6 nM, 200 μM: 193 ± 6 nM, 300 μM: 279 ± 13 nM) (Fig. 5E, Table 1). Total amount of NO released during the recording period is very stable across the subsequent days, however, the frozen stock showed a bell shape NO release curve. This resulted in a larger total amount of NO released (AUC) in comparison to fresh stock (Fig. 5F).

Bottom Line: We found that donors such as NOC-5 and PAPA-NONOate decayed substantially within days, whereas SNP and GSNO showed greater stability releasing consistent levels of NO over days.In all donors tested, the amount of released NO differs between frozen and unfrozen stocks.Fluorescent and amperometric approaches to measure NO concentrations yield a wide range of levels.

View Article: PubMed Central - PubMed

Affiliation: MRC Toxicology Unit, Hodgkin Building, University of Leicester, Leicester LE1 9HN, UK.

No MeSH data available.


Related in: MedlinePlus