Hydrogen sulfide and the vasculature: a novel vasculoprotective entity and regulator of nitric oxide bioavailability?

This is a freely-available open access publication. Please cite the published version which is available via the DOI link in this record.Hydrogen sulfide (H(2)S) is a well known and pungent toxic gas that has recently been shown to be synthesised in man from the amino acids cystathionine, homocysteine and cysteine by at least two distinct enzymes; cystathionine-gamma-lyase and cystathionine-beta-synthase. In the past few years, H(2)S has emerged as a novel and increasingly important mediator in the cardiovascular system but delineating the precise physiology and pathophysiology of H(2)S is proving to be complex and difficult to unravel with disparate findings reported with cell types, tissue types and animal species reported. Therefore, in this review we summarize the mechanisms by which H(2)S has been proposed to regulate blood pressure and cardiac function, discuss the mechanistic discrepancies reported in the literature as well as the therapeutic potential of H(2)S. We also examine the methods of H2S detection in biological fluids, processes for H(2)S removal and discuss the reported blood levels of H(2)S in man and animal models of cardiovascular pathology. We also highlight the complex interaction of H(2)S with nitric oxide in regulating cardiovascular function in health and disease

Imaging the uptake of gold nanoshells in live cells using plasmon resonance enhanced four wave mixing microscopy

This paper was published in Optics Express and is made available as an electronic reprint with the permission of OSA. The paper can be found at the following URL on the OSA website: http://dx.doi.org/10.1364/OE.19.017563. Systematic or multiple reproduction or distribution to multiple locations via electronic or other means is prohibited and is subject to penalties under law.Gold nanoshells (GNS) are novel metal nanoparticles exhibiting attractive optical properties which make them highly suitable for biophotonics applications. We present a novel investigation using plasmon-enhanced four wave mixing microscopy combined with coherent anti-Stokes Raman scattering (CARS) microscopy to visualize the distribution of 75 nm radius GNS within live cells. During a laser tolerance study we found that cells containing nanoshells could be exposed to < 2.5 mJ each with no photo-thermally induced necrosis detected, while cell death was linearly proportional to the power over this threshold. The majority of the GNS signal detected was from plasmon-enhanced four wave mixing (FWM) that we detected in the epi-direction with the incident lasers tuned to the silent region of the Raman spectrum. The cellular GNS distribution was visualized by combining the epi-detected signal with forwards-detected CARS at the CH2 resonance. The applicability of this technique to real-world nanoparticle dosing problems was demonstrated in a study of the effect of H2S on nanoshell uptake using two donor molecules, NaHS and GYY4137. As GYY4137 concentration was increased from 10 µM to 1 mM, the nanoshell pixel percentage as a function of cell volume (PPCV) increased from 2.15% to 3.77%. As NaHS concentration was increased over the same range, the nanoshell PPCV decreased from 12.67% to 11.47%. The most important factor affecting uptake in this study was found to be the rate of H2S release, with rapid-release from NaHS resulting in significantly greater uptake

Competition of reactive signals and thiol modifications of proteins

It is clear that cells are constantly bombarded by multiple signals, often initiating similar, or even conflicting, responses. Important players in this suite of signals are the reactive oxygen species (ROS), such as hydrogen peroxide (H2O2), the reactive nitrogen species (RNS), such as nitric oxide (NO) and sulfur-based molecules, such as hydrogen sulfide (H2S). These compounds are often involved in stress responses and dysfunction of these signaling systems is often involved in disease [1-3]. This commentary discusses the interactions of such signals, which was discussed in a previous paper [3]. It was argued that all these molecules are not acting in the same manner, and that H2S was acting in a role which moderated the effects of ROS and NO

Vascular Guidance: Microstructural Scaffold Patterning for Inductive Neovascularization

Current tissue engineering techniques are limited by inadequate vascularisation and perfusion of cell-scaffold constructs. Microstructural patterning through biomimetic vascular channels within a polymer scaffold might induce neovascularization, allowing fabrication of large engineered constructs. The network of vascular channels within a frontal-parietal defect in a patient, originating from the anterior branch of the middle meningeal artery, was modeled using computer-aided design (CAD) techniques and subsequently incorporated into polycaprolactone (PCL) scaffolds fabricated using fused deposition modeling (FDM). Bone marrow-derived mesenchymal stem cells (MSCs) were seeded onto the scaffolds and implanted into a rat model, with an arteriovenous bundle inserted at the proximal extent of the vascular network. After 3 weeks, scaffolds were elevated as a prefabricated composite tissue-polymer flap and transferred using microsurgical technique. Histological examination of explanted scaffolds revealed vascular ingrowth along patterned channels, with abundant capillary and connective tissue formation throughout experimental scaffolds, while control scaffolds showed only granulation tissue. All prefabricated constructs transferred as free flaps survived and were viable. We term this concept “vascular guidance,” whereby neovascularization is guided through customized channels in a scaffold. Our technique might potentially allow fabrication of much larger tissue-engineered constructs than current technologies allow, as well as allowing tailored construct fabrication with a patient-specific vessel network based on CT scan data and CAD technology

Very High-Speed Digital Video Capability for In-Flight Use

digital video camera system has been qualified for use in flight on the NASA supersonic F-15B Research Testbed aircraft. This system is capable of very-high-speed color digital imaging at flight speeds up to Mach 2. The components of this system have been ruggedized and shock-mounted in the aircraft to survive the severe pressure, temperature, and vibration of the flight environment. The system includes two synchronized camera subsystems installed in fuselage-mounted camera pods (see Figure 1). Each camera subsystem comprises a camera controller/recorder unit and a camera head. The two camera subsystems are synchronized by use of an MHub(TradeMark) synchronization unit. Each camera subsystem is capable of recording at a rate up to 10,000 pictures per second (pps). A state-of-the-art complementary metal oxide/semiconductor (CMOS) sensor in the camera head has a maximum resolution of 1,280 1,024 pixels at 1,000 pps. Exposure times of the electronic shutter of the camera range from 1/200,000 of a second to full open. The recorded images are captured in a dynamic random-access memory (DRAM) and can be downloaded directly to a personal computer or saved on a compact flash memory card. In addition to the high-rate recording of images, the system can display images in real time at 30 pps. Inter Range Instrumentation Group (IRIG) time code can be inserted into the individual camera controllers or into the M-Hub unit. The video data could also be used to obtain quantitative, three-dimensional trajectory information. The first use of this system was in support of the Space Shuttle Return to Flight effort. Data were needed to help in understanding how thermally insulating foam is shed from a space shuttle external fuel tank during launch. The cameras captured images of simulated external tank debris ejected from a fixture mounted under the centerline of the F-15B aircraft. Digital video was obtained at subsonic and supersonic flight conditions, including speeds up to Mach 2 and altitudes up to 50,000 ft (15.24 km). The digital video was used to determine the structural survivability of the debris in a real flight environment and quantify the aerodynamic trajectories of the debris

Pharmacological postconditioning against myocardial infarction with a slow-releasing hydrogen sulfide donor, GYY4137

Exogenous hydrogen sulfide (H2S) protects against myocardial ischemia/reperfusion injury but the mechanism of action is unclear. The present study investigated the effect of GYY4137, a slow-releasing H2S donor, on myocardial infarction given specifically at reperfusion and the signalling pathway involved. Thiobutabarbital-anesthetised rats were subjected to 30min of left coronary artery occlusion and 2h reperfusion. Infarct size was assessed by tetrazolium staining. In the first study, animals randomly received either no treatment or GYY4137 (26.6, 133 or 266μmolkg-1) by intravenous injection 10min before reperfusion. In a second series, involvement of PI3K and NO signalling were interrogated by concomitant administration of LY294002 or L-NAME respectively and the effects on the phosphorylation of Akt, eNOS, GSK-3β and ERK1/2 during early reperfusion were assessed by immunoblotting. GYY4137 266μmolkg-1 significantly limited infarct size by 47% compared to control hearts (P<0.01). In GYY4137-treated hearts, phosphorylation of Akt, eNOS and GSK-3β was increased 2.8, 2.2 and 2.2 fold respectively at early reperfusion. Co-administration of L-NAME and GYY4137 attenuated the cardioprotection afforded by GYY4137, associated with attenuated phosphorylation of eNOS. LY294002 totally abrogated the infarct-limiting effect of GYY4137 and inhibited Akt, eNOS and GSK-3β phosphorylation. These data are the first to demonstrate that GYY4137 protects the heart against lethal reperfusion injury through activation of PI3K/Akt signalling, with partial dependency on NO signalling and inhibition of GSK-3β during early reperfusion. H2S-based therapeutic approaches may have value as adjuncts to reperfusion in the treatment of acute myocardial infarction

AP39, A Mitochondrially- Targeted Hydrogen Sulfide Donor, Exerts Protective Effects in Renal Epithelial Cells Subjected to Oxidative Stress in Vitro and in Acute Renal Injury in Vivo.

This is the author's accepted manuscript. The article is published in final form in Shock, DOI: 10.1097/SHK.0000000000000478This study evaluated the effects of AP39 [(10-oxo-10-(4-(3-thioxo-3H-1,2-dithiol-5yl) phenoxy)decyl) triphenyl phosphonium bromide], a mitochondrially targeted donor of hydrogen sulfide (H2S) in an in vitro model of hypoxia/oxidative stress injury in NRK-49F rat kidney epithelial cells (NRK cells) and in a rat model of renal ischemia-reperfusion injury. Renal oxidative stress was induced by the addition of glucose oxidase, which generates hydrogen peroxide in the culture medium at a constant rate. Glucose oxidase (GOx)-induced oxidative stress led to mitochondrial dysfunction, decreased intracellular ATP content, and, at higher concentrations, increased intracellular oxidant formation (estimated by the fluorescent probe 2, 7-dichlorofluorescein, DCF) and promoted necrosis (estimated by the measurement of lactate dehydrogenase release into the medium) of the NRK cells in vitro. Pretreatment with AP39 (30-300 nM) exerted a concentration-dependent protective effect against all of the above effects of GOx. Most of the effects of AP39 followed a bell-shaped concentration-response curve; at the highest concentration of GOx tested, AP39 was no longer able to afford cytoprotective effects. Rats subjected to renal ischemia/reperfusion responded with a marked increase (over 4-fold over sham control baseline) blood urea nitrogen and creatinine levels in blood, indicative of significant renal damage. This was associated with increased neutrophil infiltration into the kidneys (assessed by the myeloperoxidase assay in kidney homogenates), increased oxidative stress (assessed by the malondialdehyde assay in kidney homogenates) and an increase in plasma levels of IL-12. Pretreatment with AP39 (0.1, 0.2 and 0.3 mg/kg) provided a dose-dependent protection against these pathophysiological alterations; the most pronounced protective effect was observed at the 0.3 mg/kg dose of the H2S donor; nevertheless AP39 failed to achieve a complete normalization of any of the injury markers measured. The partial protective effects of AP39 correlated with a partial improvement of kidney histological scores and reduced TUNEL staining (an indicator of DNA damage and apoptosis). In summary, the mitochondria-targeted H2S donor AP39 exerted dose-dependent protective effects against renal epithelial cell injury in vitro and renal ischemia-reperfusion injury in vivo. We hypothesize that the beneficial actions of AP39 are related to the reduction of cellular oxidative stress, and subsequent attenuation of various positive feed-forward cycles of inflammatory and oxidative processes.National Institutes of Healt

Mitochondria-targeted hydrogen sulfide donor AP39 improves neurological outcomes after cardiac arrest in mice

Copyright © 2015 Elsevier. NOTICE: this is the author’s version of a work that was accepted for publication in . Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Nitric Oxide, Vol. 49, pp. 90–96 (2015), DOI:10.1016/j.niox.2015.05.001Aims Mitochondria-targeted hydrogen sulfide donor AP39, [(10-oxo-10-(4-(3-thioxo-3H-1,2-dithiol-5yl)phenoxy)decyl) triphenylphosphonium bromide], exhibits cytoprotective effects against oxidative stress in vitro. We examined whether or not AP39 improves the neurological function and long term survival in mice subjected to cardiac arrest (CA) and cardiopulmonary resuscitation (CPR). Methods Adult C57BL/6 male mice were subjected to 8 min of CA and subsequent CPR. We examined the effects of AP39 (10, 100, 1000 nmol kg−1) or vehicle administered intravenously at 2 min before CPR (Experiment 1). Systemic oxidative stress levels, mitochondrial permeability transition, and histological brain injury were assessed. We also examined the effects of AP39 (10, 1000 nmol kg−1) or vehicle administered intravenously at 1 min after return of spontaneous circulation (ROSC) (Experiment 2). ROSC was defined as the return of sinus rhythm with a mean arterial pressure >40 mm Hg lasting at least 10 seconds. Results Vehicle treated mice subjected to CA/CPR had poor neurological function and 10-day survival rate (Experiment 1; 15%, Experiment 2; 23%). Administration of AP39 (100 and 1000 nmol kg−1) 2 min before CPR significantly improved the neurological function and 10-day survival rate (54% and 62%, respectively) after CA/CPR. Administration of AP39 before CPR attenuated mitochondrial permeability transition pore opening, reactive oxygen species generation, and neuronal degeneration after CA/CPR. Administration of AP39 1 min after ROSC at 10 nmol kg−1, but not at 1000 nmol kg−1, significantly improved the neurological function and 10-day survival rate (69%) after CA/CPR. Conclusion The current results suggest that administration of mitochondria-targeted sulfide donor AP39 at the time of CPR or after ROSC improves the neurological function and long term survival rates after CA/CPR by maintaining mitochondrial integrity and reducing oxidative stress.National Institutes of Healt

Gendered violence and sexualized representations in video games: (Lack of) effect on gender-related attitudes

This research explored how gender portrayals in video games affect gender-related attitudes. Two hundred participants from the United Kingdom and Malaysia participated across three experiments, where the appearance and behaviour of video game characters were manipulated with regard to target (enemy) gender (Study 1), sexually explicit attire (Study 2) and level of character agency (Study 3). We found minimal evidence that exposure to gender-stereotyped content resulted in differential gender-related attitudes (implicit associations, hostile and benevolent sexism, or rape myth acceptance). However, Study 1 findings showed that individuals who played a first-person shooter with male enemies showed lower endorsement of some (benevolent) sexist attitudes (cf. control) and showed difference in game behaviour (cf. female enemies). Together, our results suggest that short-term exposure to video games containing female characters (sexualised, passive, or otherwise) does not consistently lead to the endorsement of negative gender attitudes

Ap39, a mitochondria-targeting hydrogen sulfide (H2 s) donor, protects against myocardial reperfusion injury independently of salvage kinase signalling

Background and Purpose H2S protects myocardium against ischaemia-reperfusion injury. This protection may involve the cytosolic reperfusion injury salvage kinase (RISK) pathway, but direct effects on mitochondrial function are possible. Here, we investigated the potential cardioprotective effect of mitochondria-specific H2S donor, AP39, at reperfusion against ischaemia/reperfusion injury. Experimental Approach Anaesthetised rats underwent myocardial (30 min ischaemia/120 min reperfusion) with randomisation to receive interventions prior to reperfusion: vehicle, AP39 (0.01, 0.1, 1 µmol kg-1), or the control compounds AP219 or ADT-OH (1 µmol kg-1). LY294002, L-NAME or ODQ were used to interrogate the involvement of RISK pathway. Myocardial samples harvested 5 minutes after reperfusion were analysed for RISK protein phosphorylation and additional experiments were conducted on isolated cardiac mitochondria to examine the direct mitochondrial effects of AP39. Key Results AP39 exerted dose-dependent infarct size limitation. Inhibition of either PI3K/Akt, eNOS or sGC did not affect the infarct limitation of AP39. Western blot analysis confirmed that AP39 did not induce phosphorylation of Akt, eNOS, GSK-3β or ERK1/2. In isolated subsarcolemmal and interfibrillar mitochondria, AP39 significantly attenuated mitochondrial ROS generation without affecting respiratory complexes I or II. Further, AP39 inhibited mitochondrial permeability transition pore (PTP) opening and co-incubation of mitochondria with AP39 and cyclosporine A induced an additive inhibition of PTP. Conclusion and Implications AP39 protects against reperfusion injury independently of the cytosolic RISK pathway. Cardioprotection could be mediated by inhibiting PTP via cyclophilin D-independent mechanism. Thus, selective delivery of H2S to mitochondria may be therapeutically applicable for harnessing the cardioprotective utility of H2S. This article is protected by copyright. All rights reserved