Photochemical Release Of Nitric Oxide Research Articles

Mechanistic analysis of the photolytic decomposition of solid-state S-nitroso-N-acetylpenicillamine

S-Nitroso-N-acetylpenicillamine (SNAP) is among the most common nitric oxide (NO)-donor molecules and its solid-state photolytic decomposition has potential for inhaled nitric oxide (iNO) therapy. The photochemical NO release kinetics and mechanism were investigated by exposing solid-state SNAP to a narrow-band LED as a function of nominal wavelength and intensity of incident light. The photolytic efficiency, decomposition products, and the photolytic pathways of the SNAP were examined. The maximum light penetration depth through the solid layer of SNAP was determined by an optical microscope and found to be within 100–200 μm, depending on the wavelength of light. The photolysis of solid-state SNAP to generate NO along with the stable thiyl (RS·) radical was confirmed using Electron Spin Resonance (ESR) spectroscopy. The fate of the RS· radical in the solid phase was studied both in the presence and absence of O2 using NMR, IR, ESR, and UPLC-MS. The changes in the morphology of SNAP due to its photolysis were examined using PXRD and SEM. The stable thiyl radical formed from the photolysis of solid SNAP was found to be reactive with another adjacent thiyl radical to form a disulfide (RSSR) or with oxygen to form various sulfonyl and sulfonyl peroxyl radicals {RS(O)xO·, x = 0 to 7}. However, the thiyl radical did not recombine with NO to reform the SNAP. From the PXRD data, it was found that the SNAP loses its crystallinity by generating the NO after photolysis. The initial release of NO during photolysis was increased with increased intensity of light, whereas the maximum light penetration depth was unaffected by light intensity. The knowledge gained about the photochemical reactions of SNAP may provide important insight in designing portable photoinduced NO-releasing devices for iNO therapy.

Photodissociation Dynamics of Nitric Oxide from N-Acetylcysteine- or N-Acetylpenicillamine-bound Roussin's Red Ester.

The photochemical release of nitric oxide (NO) from a NO precursor is advantageous in terms of spatial, temporal, and dosage control of NO delivery to target sites. To realize full control of the quantitative NO administration from photoactivated NO precursors, it is necessary to have detailed dynamical information on the photodissociation of NO from NO precursors. We synthesized two new water-soluble Roussin’s red esters (RREs), [Fe2(μ-N-acetylcysteine)2(NO)4] and [Fe2(μ-N-acetylpenicillamine)2(NO)4], which have five times longer lifetime than the well-known [Fe2(μ-cysteine)2(NO)4]. The photodissociation dynamics of NO from these RREs in water were investigated over a broad time range from 0.3 ps to 10 μs after excitation at 310 and 400 nm using femtosecond time-resolved infrared (IR) spectroscopy. When these RREs are excited, they either release one NO, producing a radical species deficient in one NO (R), [Fe2(μ-RS)2(NO)3], or relax into the ground state without photodeligation via an electronically excited intermediate state (M). R appears immediately after photoexcitation, suggesting that one NO is photodissociated faster than 0.3 ps. A certain fraction of R undergoes geminate recombination (GR) with NO with a time constant of 7–9 ps, while the remaining R competitively binds to the solvent. Solvent-bound R eventually bimolecularly recombines with NO with a rate constant of (1.3–1.6) × 108 M–1 s–1. For a given RRE molecule, the fractional yield of M (0.62–0.76) depends on the excitation wavelength (λex); however, the relaxation time of M (6 ± 1 ns) is independent of λex. Although the primary quantum yield of NO photodissociation (Φ1) was found to be 0.24–0.38, the final yield of NO suitable for other reactions (Φ2) was reduced to 0.14–0.29 due to the picosecond GR of the dissociated NO with R. Detailed photoexcitation dynamics of RRE can be utilized in the quantitative control of NO administration at a specific site and time, promoting pin-point usage of NO in chemistry and biology. We demonstrate that femtosecond IR spectroscopy combined with quantum chemical calculations is a powerful method for obtaining detailed dynamic information on photoactivated NO precursors such as Φ1 and Φ2, the GR yield, and secondary reactions of the nascent photoproducts, which are essential information for the design of efficient photoactivated NO precursors and their quantitative utilization.

Influence of Pluronic F127 microenvironments on the photochemical nitric oxide release from S-nitrosoglutathione

Poly(ethylene oxide)-poly(propylene oxide)-poly(ethylene oxide) (PEO-PPO-PEO) (F127) hydrogels have been used to deliver nitric oxide (NO) topically in biomedical applications. Here, the effect of F127 microenvironments on the photochemical NO release from S-nitrosoglutathione (GSNO) was investigated in F127 solutions 7.6 wt% 15 wt% and 22.5 wt% at 15 °C and 37 °C. Small-angle X-ray Scattering (SAXS) and Differential Scanning Calorimetry (DSC) measurements, along with proton Nuclear Magnetic Resonance (1H NMR) spectral shifts and T2 relaxation data at six different concentration-temperature conditions, allowed identifying F127 microphases characterized by: a sol phase of unimers; micelles in non-defined periodic order, and a gel phase of cubic packed micelles. Kinetic measurements showed that GSNO photodecompositon proceeds faster in micellized F127 where GSNO is segregated to the intermicellar microenvironment. Real time kinetic monitoring of NO release and T2 relaxation profiles showed that NO is preferentially partitioned into the hydrophobic PPO cores of the F127 micelles, with the consequent decrease in its rate of release to the gas phase. These results show that F127 microphases affect both the kinetics of GSNO photodecomposition and the rate of NO escape and can be used to modulate the photochemical NO delivery from F127/GSNO solutions.

Topical photochemical nitric oxide release from porous poly(vinyl alcohol) membrane for visible light modulation of dermal vasodilation

Abstract Complications of functional peripheral artery disease (PAD), like chronic wounds and Raynaud’s disease and phenomenon, have stimulated the development of topical nitric oxide (NO)-releasing biomaterials capable of restoring the blood flow locally. In this work, we used phase inversion technique to prepare porous membranes of poly(vinyl alcohol) (PVA) functionalized with NO-releasing S-nitrosothiol (SNO) moieties. This strategy led to crosslinked amorphous membranes with swelling degree of 750 wt% and interconnected macropores with diameters greater than 600 μm. Controlling the extent of the S-nitrosation reaction allowed obtaining PVA-SNO membranes that spontaneously release NO in the range of 33–53 nmol/cm 2 in the dark. Irradiation of these membranes with visible light, led to first order NO release kinetics with rate constant of (5.6 ± 0.2) × 10 −3 s −1 and increased the amount of NO released to 64–101 nmol/cm 2 . Topical application of PVA-SNO membrane for 4 min on the healthy skin, led to a fast and localized 300% increase in the dermal blood flow, which was enhanced to 900% when the membrane was irradiated with visible light, during the same period. These results show that the photochemical NO release from macroporous PVA-SNO membrane is an effective tool for modulating NO actions in the treatment/prevention of PAD symptoms and other biomedical applications.

Light-controlled release of nitric oxide from solid polymer composite materials using visible and near infra-red light.

Photochemical Nitric oxide releasing composite materials (Photo-NORMs) were prepared using biocompatible polymers and the photochemical nitric oxide donor complex (CrONO). We have demonstrated nitric oxide (NO) release from the solid composites for extended (>30 hours) and controlled (20-100 pmoles s(-1)) durations after visible light irradiation. Quantitation of the efficiency of NO release from the composites shows that polymer gas permeability most dramatically affects the overall efficiency (QY) of photochemical NO release, where polymers with higher gas permeability have a higher QY of nitric oxide release. Composites were also prepared with β-phase lanthanide-doped NaYF4 upconverting nanoparticles (UCNPs). Controlled Nitric oxide release was achieved via near infrared (NIR) irradiation. A prototype LED device shows proof-of-concept that such photoresponsive NO-releasing composites could be applied to implantable systems, where the amount of NO released is modulated by changing irradiation time and light intensity. This research provides the guidelines necessary to move towards device fabrication and testing in actual tissue to evaluate the photo-NORMS as a reliable option for nitric oxide release in vivo.

Photochemical and Electrochemical Study of the Release of Nitric Oxide from [Ru(bpy)2L(NO)](PF6)nComplexes (L = Imidazole, 1-Methylimidazole, Sulfite and Thiourea), Toward the Development of Therapeutic Photodynamic Agents

The development of NO photoreleaser compounds has important potential applications on medicine, particularly on preventing topic infections and controlling cancers. Due to these expectations, the photochemical release of nitric oxide from complexes of [Ru(bpy)2LX]n+, where L = imidazole, 1-methylimidazole, sulphite and thiourea and X = NO+ and NO2− was investigated employing spectroscopic and electrochemical techniques. The release of NO was confirmed by chronoamperometry using a NO selective electrode, while the other product, mainly [RuIIH2O], was detected by UV-Visible spectroscopy and electrochemical techniques for all complexes except for thiourea. The amount of NO released by these complexes upon irradiation was determined using a new developed method using square wave voltammetry.

Enhanced photochemical nitric oxide release from a flutamide derivative incorporated in Pluronic F127 micelles

The incorporation of nitric oxide (NO) photodonors into nontoxic vehicles may allow for the tunable control of topical NO release to treat pathological conditions associated with the impairment of NO production in the dermal vasculature. Pluronic F127 is a PEO–PPO–PEO triblock copolymer that is used as a nontoxic vehicle that undergoes thermally reversible micellization and gelation. In this study, we synthesized a thermally stable flutamide derivative, N-(3-aminopropyl)-3-(trifluoromethyl)-4-nitrobenzenamine (ATN), which is capable of releasing NO upon irradiation with visible light, as shown by real-time chemiluminescence NO detection. We demonstrated that the incorporation of ATN into the hydrophobic nuclei of F127 micelles leads to an enhancement of its photochemical NO release, which is consistent with the photodecomposition route associated with the nitro-to-nitrite photorearrangement of ATN. This mechanism underscores a new role of the F127 micelles as concentrating “nanoreactors” for hydrophobic NO releasing drugs. In this case, this effect may represent a new strategy for enhancing photochemical NO release from ATN, which makes the micellar F127/ATN solutions promising for topical NO application.

Detailed mechanism of the autoxidation of N-hydroxyurea catalyzed by a superoxide dismutase mimic Mn(iii) porphyrin: formation of the nitrosylated Mn(ii) porphyrin as an intermediate

The in vitro autoxidation of N-hydroxyurea (HU) is catalyzed by Mn(III)TTEG-2-PyP(5+), a synthetic water soluble Mn(III) porphyrin which is also a potent mimic of the enzyme superoxide dismutase. The detailed mechanism of the reaction is deduced from kinetic studies under basic conditions mostly based on data measured at pH = 11.7 but also including some pH-dependent observations in the pH range 9-13. The major intermediates were identified by UV-vis spectroscopy and electrospray ionization mass spectrometry. The reaction starts with a fast axial coordination of HU to the metal center of Mn(III)TTEG-2-PyP(5+), which is followed by a ligand-to-metal electron transfer to get Mn(II)TTEG-2-PyP(4+) and the free radical derived from HU (HU˙). Nitric oxide (NO) and nitroxyl (HNO) are minor intermediates. The major pathway for the formation of the most significant intermediate, the {MnNO} complex of Mn(II)TTEG-2-PyP(4+), is the reaction of Mn(II)TTEG-2-PyP(4+) with NO. We have confirmed that the autoxidation of the intermediates opens alternative reaction channels, and the process finally yields NO(2)(-) and the initial Mn(III)TTEG-2-PyP(5+). The photochemical release of NO from the {MnNO} intermediate was also studied. Kinetic simulations were performed to validate the deduced rate constants. The investigated reaction has medical implications: the accelerated production of NO and HNO from HU may be utilized for therapeutic purposes.

Visible Light‐Induced Release of Nitrogen Monoxide from a Nitrosylrhodium Complex

The important roles that nitric oxide (NO) plays in biological environments, and the need for precise and targeted delivery of NO for medicinal and other purposes have led to intense research in the area of metal nitrosyl complexes as thermal and photochemical sources of NO. Complexes with a good combination of chemical stability and high quantum yield for photochemical release of NO upon irradiation with visible light in aqueous solutions are rare. Here we report that a simple macrocyclic nitrosylrhodium complex [L(2)(H(2)O)Rh(NO)](2+) (L(2)=Me(6)[14]aneN(4)) exhibits unique chemical and photochemical properties that make it an excellent photochemical precursor of NO. The complex is highly soluble in water, thermally stable, and resistant toward O(2). Irradiation in the 648 nm band generates NO and [L(2)(H(2)O)Rh](2+) in aqueous solutions with a quantum yield of 1.00±0.07, the highest ever reported for a nitrosyl complex under any conditions. In the absence of O(2), the two fragments combine to regenerate [L(2)(H(2)O)Rh- (NO)](2+), but in O(2)-containing solutions, [L(2)(H(2)O)RhOO](2+) is formed as determined in spectral and kinetic measurements. The kinetics of the reaction of this superoxo complex with NO were measured by laser flash photolysis, k=(3.9±0.4)×10(7) M(-1) s(-1). Steady-state photolysis of [L(2)(H(2)O)Rh(NO)](2+) under O(2) yielded [L(2)(H(2)O)Rh(ONO(2))](2+), a long-lived nitrato intermediate that can also be generated in a direct reaction between NO and genuine [L(2)(H(2)O)RhOO](2+). Thus, visible-light photolysis of the [L(2)(H(2)O)Rh(NO)](2+)/O(2) system converts it to the [L(2)(H(2)O)RhOO](2+)/NO combination.

Decomposition of S‐Nitrosothiols Induced by UV and Sunlight

Photochemical release of nitric oxide (NO) from the S‐nitroso derivatives of glutathione, L‐cysteine, N‐acetyl‐L‐cysteine, L‐cysteinemethylester, D,L‐penicillamine, N‐acetyl‐D,L‐penicillamine, and N‐acetylcysteamine has been investigated at neutral and acidic pH. The release of NO from RSNO is one of the key reactions that could be utilized in photodynamic therapy. The UV‐VIS and HPLC analyses have shown that under argon saturated conditions, disulfide (RSSR) is the major product of UV as well as sunlight induced decomposition. While in aerated conditions, nitirite—the end product of the oxidation of NO—was also observed along with disulfide. The formation of thiyl radical as the intermediate was reconfirmed by laser flash photolysis. The initial rate of formation of NO was on the order of 10−5 dm3 mol−1 s−1. The quantum yields of these reactions were in the range of 0.2–0.8. The high quantum yields observed in the photo induced release of NO from RSNO using both UV and sunlight demonstrate the potential application of these reactions in photodynamic therapy.

Photochemical NO release from nitrosyl Ru II complexes with C-bound imidazoles

The series of nitrosyl complexes trans-[Ru(NH 3) 4L(NO)]Cl 3, L = caffeine, theophylline, imidazole and benzoimidazole in position trans to NO were prepared and their photochemical properties studied. All complexes showed nitric oxide (NO) release under light irradiation at 330–440 nm. Quantum yields for [Ru(NH 3) 4L(H 2O)] 3+ formation ( ϕ Ru(III)) were sensitive to the natures of L, λ irr and pH. The major product of the irradiation of trans-[Ru(NH 3) 4L(NO +)] 3+ is the trans-[Ru III(NH 3) 4L(Cl)] 2+ and NO as suggested by UV–Vis, electrochemical, and FTIR techniques.

Water-Soluble Two-Photon Absorbing Nitrosyl Complex for Light-Activated Therapy through Nitric Oxide Release

A water-soluble nitrosyl complex with large two-photon absorption was synthesized by incorporating a two-photon absorbing chromophore with tetra(ethylene glycol) units, into the Roussin's red salt. The nitrosyl complex exhibits quenched emission due to energy transfer from the two-photon chromophore to the Roussin's red salt. The nitric oxide (NO) release induced by one- or two-photon irradiation was detected by EPR spectroscopy with a chemical probe, the Fe(II)- N-(dithiocarbamoyl)- N-methyl- d-glucamine (Fe-MGD) complex. Increased one- or two-photon excited fluorescence, with a concomitant photochemical release of NO, was observed upon one- or two-photon light irradiation. With the observed light-dependent cytotoxicity against cancer cells of the water-soluble nitrosyl complex, it was demonstrated that two-photon-functionalized nitrosyl complexes can be effective NO donors for light-activated treatment.

P045. Local blue laser irradiation improves tissue circulation by photochemical release of nitric oxide from nitrosyl hemoglobin

P045. Local blue laser irradiation improves tissue circulation by photochemical release of nitric oxide from nitrosyl hemoglobin

Photochemical release of nitric oxide from a regenerable, sol-gel encapsulated Ru–salen–nitrosyl complex

Light activation leads to release of NO from a silicate sol-gel material SG-RuNO prepared from the ruthenium complex, [Ru(salen)(OH2)(NO)]+ (salen = N,N'-bis-(salicylidene)ethyl-enediaminato); after photochemical NO photolabilization, SG-RuNO can be regenerated from the spent material via the subsequent reaction with aqueous nitrite.

S-Nitrosoglutathione incorporated in poly(ethylene glycol) matrix: potential use for topical nitric oxide delivery

Incorporation of nitric oxide (NO) donors in non-toxic polymeric matrices can be a useful strategy for allowing topical NO delivery. We have incorporated the NO-donor S-nitrosoglutathione (GSNO) into a liquid poly(ethylene glycol) (PEG)/H 2O matrix through the S-nitrosation of GSH by a NO/O 2 gas mixture. Kinetic measurements of GSNO decomposition associated with NO release were performed at 25, 35, and 45 °C in the dark and under irradiation with UV/Vis light, λ > 480 nm and λ = 333 nm. NO release from the liquid matrix to the gas phase was confirmed by mass spectrometry. The PEG/H 2O matrix stabilizes GSNO leading to expressive reductions in the initial rates of thermal and photochemical NO release, compared to aqueous GSNO solution. This matrix effect is assigned to diffusional constrains imposed on the escape of the NO and GS radicals formed in the solvent cage. This effect allows the storage of PEG-GSNO formulations for extended periods (more than 65 days at freezer) with negligible decomposition. PEG-GSNO formulation seems therefore to be applicable in topical NO delivery and GSNO displays potential as a percutaneous absorption enhancer. Moreover, the rate of NO release can be locally increased by irradiation with visible light.

Thermal and photochemical nitric oxide release from S-nitrosothiols incorporated in Pluronic F127 gel: potential uses for local and controlled nitric oxide release

The local delivery of nitric oxide (nitrogen monoxide, NO) by thermal or photochemical means to target cells or organs has a great potential in several biomedical applications, especially if the NO donors are incorporated into non-toxic viscous matrices. In this work, we have shown that the NO donors S-nitrosoglutathione (GSNO) and S-nitroso- N-acetylcysteine (SNAC) can be incorporated into F127 hydrogels, from where NO can be released thermally or photochemically (with λ irr>480 nm). High sensitivity differential scanning calorimetry (HSDSC) and a new spectrophotometric method, were used to characterize the micellization and the reversal thermal gelation processes of the F127 hydrogels containing NO donors, and to modulate the gelation temperatures to the range 29–32°C. Spectral monitoring of the S–NO bond cleavage showed that the initial rates of thermal and photochemical NO release (ranging from 2 to 45 μmol l −1 min −1) are decreased in the hydrogel matrices, relative to those obtained in aqueous solutions. This stabilization effect was assigned to a cage recombination mechanism and offers an additional advantage for the storage and handling of S-nitrosothiols. These results indicate that F127 hydrogels might be used for the thermal and photochemical delivery of NO from S-nitrosothiols to target areas in biomedical applications.

Controlled Photochemical Release of Nitric Oxide from O2-Naphthylmethyl- and O2-Naphthylallyl-Substituted Diazeniumdiolates

The photochemistry of O2-naphthylmethyl- and O2-naphthylallyl-substituted diazeniumdiolates has been investigated. Electron-donating methoxy group substitution is shown to have a significant effect on the observed photochemistry, with the appropriate substitution pattern resulting in efficient diazeniumdiolate photorelease. Observed nitric oxide release rates from these photoprecursors are consistent with those expected for normal thermal dissociation of the diazeniumdiolate in aqueous solutions and show the same pH dependence.

Controlled photochemical release of nitric oxide from O2-benzyl-substituted diazeniumdiolates.

An investigation of potential photosensitive protecting groups for diazeniumdiolates (R2N-N(O)=NO-) has been initiated, and here the effect of meta electron-donating groups on the photochemistry of O2-benzyl-substituted diazeniumdiolates (R2N-N(O)=NOCH2Ar) is reported. Photolysis of the parent benzyl derivative (Ar = Ph) results almost exclusively in undesired photochemistry-the formation of nitrosamine and an oxynitrene intermediate with very little, if any, photorelease of the diazeniumdiolate. We have been able to use meta substitution to tune the photochemistry of these benzylic systems. The desired diazeniumdiolate photorelease has been shown to become more substantial with stronger pi-donating meta substituents. This effect has been verified by direct observation of the photoreleased diazeniumdiolate with 1H NMR spectroscopy and by NO quantification measurements conducted in high- and low-pH solutions. In addition, the observed rates of NO release are consistent with that expected for normal thermal decomposition of the diazeniumdiolate in aqueous solutions and also show the same pH dependence.

The Mechanism of Photochemical Release of Nitric Oxide from S‐Nitrosoglutathione

Abstract— Laser flash photolysis of S‐nitroso complexes of glutathione (GSNO) and bovine serum albumin (BSANO) via excitation at 355 nm has been used to investigate the photogeneration of nitric oxide (NO) and subsequent radical reactions. In the case of GSNO, liberation of NO was confirmed by its oxidation of oxyhemoglobin to met hemoglobin. Initial NO release is via homolytic cleavage of the S‐N bond to produce the glutathione thiyl radical, GS, which can subsequently react with (a) ground‐state GSNO (k= 1.7 × 109M−1/i> s−1) to yield additional NO and oxidized glutathione, GSSG; and (b) oxygen (k= 3.0 × 109M−1 s−1) to give the glutathione peroxy radical, GSOO, which subsequently reacts with ground‐state GSNO (k= 3.8 × 108M−1 s−1), also producing additional NO and GSSG. The relative concentrations of oxygen and GSNO in the system determine the major pathway for removal of G'. These secondary reactions occur at such high rates that they preclude radical recombination under low‐intensity irradiation conditions. The quantum yield of overall loss of GSNO thus varies with both GSNO and oxygen concentrations; a value of 0.66 was determined for an aerated solution of GSNO (0.86 mM). In the case of GSNO, therefore, generation of NO is not due solely to homolysis of the S‐N bond; secondary reactions of the radicals formed lead to further NO liberation. In rationalizing the known phototoxicity of GSNO, possible contributions from thiyl and thiyl‐derived radicals should be considered. In contrast to GSNO, direct excitation of BSANO (containing one bound NO group per molecule) led to photodecomposition with a quantum yield of 0.09 but no evidence was obtained for liberation of NO into the bulk medium.

Vasodilator responses of rat isolated tail artery enhanced by oxygen‐dependent, photochemical release of nitric oxide from iron‐sulphur‐nitrosyls

1. The vasodilator properties and photochemical decomposition of two synthetic iron-sulphur-nitrosyl clusters (cluster A: [Fe4S4(NO)4], tetranitrosyl-tetra-mu 3-sulphido-tetrahedro-tetrairon; and B:[Fe4S3 (NO)7]-1, heptanitrosyl-tri-mu 3-thioxotetraferrate(-1)) have been investigated. Experiments were carried out on isolated, internally-perfused segments of rat tail artery. 2. Bolus injections (10 microliters) of A or B ( > 0.25 mM) delivered into the internal perfusate generated sustained (or S-type) vasodilator responses, characterized by a persistent plateau of reduced tone due to NO released from clusters which enter and become trapped within endothelial cells. Clusters were therefore irradiated with visible laser light (lambda = 457.9 or 514.5 nm) either (a) in solution, while passing through a glass tube en route to the artery; or (b) when retained within the endothelium, by illuminating the artery directly during the plateau of an S-type response. Irradiation produced an additional vasodilator response, the magnitude of which depended upon wavelength and laser beam energy. 3. The nitric oxide synthase inhibitor, NG-monomethyl-L-arginine (100 microM), had no effect on light-induced vasodilator responses. However, they were (a) blocked entirely by adding oxyhaemoglobin (5 microM) to the internal perfusate; and (b) greatly enhanced by the enzyme superoxide dismutase (150 u ml-1). 4. Photolysis of cluster B was measured by absorption spectroscopy and by detecting NO released with an electrochemical sensor. The photochemical reaction was found to be oxygen-dependent. The half-time for inactivation of cluster-derived NO was measured by interposing different lengths of tubing (i.e. time delays) between the photolysis tube and NO sensor. The steady-state probe current decayed exponentially with increasing delay time, with a t 1/2 of 21 s. The amplitudes of vasodilator responses of the tail artery also decreased exponentially by increasing the time delay (t 1/2 = 58 s). Superoxide dismutase (150 u ml-1) prevented this from happening, showing that "inactivation' of cluster-derived NO was caused by reaction with superoxide anions formed during photolysis. 5. We conclude that potentiation of vasodilator responses to iron-sulphur-nitrosyl clusters by visible light is due to an oxygen-dependent photochemical reaction which accelerates the release of ligated nitrosyl groups as free NO. Based on our measurements, we estimate that ca 100 pM NO is sufficient to produce a just-detectable additional vasodilatation and that the ED50 dose is ca 3.7 nM.