Free H2S Research Articles

Detection of changes in mitochondrial hydrogen sulfide i n vivo in the fish model Poecilia mexicana (Poeciliidae).

ABSTRACTIn this paper, we outline the use of a mitochondria-targeted ratiometric mass spectrometry probe, MitoA, to detect in vivo changes in mitochondrial hydrogen sulfide (H2S) in Poecilia mexicana (family Poeciliidae). MitoA is introduced via intraperitoneal injection into the animal and is taken up by mitochondria, where it reacts with H2S to form the product MitoN. The MitoN/MitoA ratio can be used to assess relative changes in the amounts of mitochondrial H2S produced over time. We describe the use of MitoA in the fish species P. mexicana to illustrate the steps for adopting the use of MitoA in a new organism, including extraction and purification of MitoA and MitoN from tissues followed by tandem mass spectrometry. In this proof-of-concept study we exposed H2S tolerant P. mexicana to 59 µM free H2S for 5 h, which resulted in increased MitoN/MitoA in brain and gills, but not in liver or muscle, demonstrating increased mitochondrial H2S levels in select tissues following whole-animal H2S exposure. This is the first time that accumulation of H2S has been observed in vivo during whole-animal exposure to free H2S using MitoA. This article has an associated First Person interview with the first author of the paper.

Mitigation of Extreme Ocean Anoxic Event Conditions by Organic Matter Sulfurization

AbstractPast occurrences of widespread and severe anoxia in the ocean have frequently been associated with abundant geological evidence for free hydrogen sulfide (H2S) in the water column, so‐called euxinic conditions. Free H2S may react with, and modify, the chemical structure of organic matter settling through the water column and in marine sediments, with hypothesized implications for carbon sequestration. Here, taking the example of Ocean Anoxic Event 2, we explore the potential impact of organic matter sulfurization on marine carbon and oxygen cycling by means of Earth system modeling. Our model experiments demonstrate that rapid sulfurization (ksulf≥ = 105 M−1 year−1) of organic matter in the water column can drive a more than 30% enhancement of organic carbon preservation and burial in marine sediments and hence help accelerate climate cooling and Ocean Anoxic Event 2 recovery. As a consequence of organic matter sulfurization, we also find that H2S can be rapidly scavenged and the euxinic ocean volume reduced by up to 80%—helping reoxygenate the ocean as well as reducing toxic H2S emissions to the atmosphere, with potential implications for the kill mechanism at the end‐Permian. Finally, we find that the addition of organic matter sulfurization induces a series of additional feedbacks, including further atmospheric CO2 drawdown and ocean reoxygenation by the creation of a previously unrecognized net source of alkalinity to the ocean as H2S is scavenged and buried.

Gas chromatographic-sulfur chemiluminescent detector procedures for the simultaneous determination of free forms of volatile sulfur compounds including sulfur dioxide and for the determination of their metal-complexed forms

Three different procedures for the quantitative assessment of free and metal complexed volatile sulfur compounds (VSCs) and for the determination of truly free SO2 have been developed, taking advantage of a GC-sulfur chemiluminescent detector system (GC-SCD) with cryotrapping. The inertness of the inlet systems, together with the column used (SPB-1 sulfur) makes it possible to obtain a non-saturated perfectly Gaussian peak for SO2, well resolved from H2S. In the main procedure, the injection of 1 mL of the headspace of a sample prepared in complete anoxia and equilibrated at 30 °C makes it possible to get highly sensitive signals for all VSCs and free SO2. Detection limits are 3, 35 and 60 ng/L for H2S, MeSH and EtSH, 13 μg/L for truly free SO2 (at pH = 3.4, or 0.46 μg/L for molecular SO2), and better than 1 μg/L for other relevant sulfur volatiles. Method precision is also satisfactory and linearity covers the whole range of occurrence of these compounds. A second procedure, not making use of the cryotrapping unit, gives also satisfactory results, although with higher detection limits (0.03, 0.25 and 0.37 μg/L for free H2S, MeSH and EtSH, respectively). For the analysis of free plus metal-complexed forms, it has been demonstrated that the headspace injection of the vapors on a 1:10 brine dilution of the sample heated at 70 °C for 25 min, gives good estimates of the free + metal-complexed forms of H2S and wine mercaptans.

Petrologic and geochemical evidence for the formation of organic-rich siliceous rocks of the Late Permian Dalong Formation, Lower Yangtze region, southern China

Abstract Black shales are economically and scientifically important because they are commonly associated with high-quality hydrocarbon source rocks and metal accumulations formed during periods of abrupt environmental change. Organic-rich siliceous rocks of the Late Permian Dalong Formation developed extensively within Middle–Upper Yangtze region (western Yangtze platform); however, the formation mechanism of its formation in the Lower Yangtze region (eastern Yangtze platform) remains a subject of debate due to a lack of outcrop exposure. Petrologic, mineralogic, and geochemical data are presented for three sections of the Dalong Formation within the Lower Yangtze region. Geochemical ratios including V/(V + Ni), V/Cr, Ni/Co, U/Th, and V/Sc suggest that the Dalong Formation was deposited in an oxygen-poor environment. The elevated concentration of Mo (>100 ppm) further suggests that free H2S existed in the bottom water, which was favorable for the accumulation of large amounts of organic matter. Mo/TOC ratios reveal that water flow in the Lower Yangtze Basin was moderately to strongly restricted during deposition of the Dalong Formation. Flow within the upper–middle parts of the Yangtze Basin was also restricted, suggesting that northern marginal basins on the Yangtze Platform formed part of a single ocean during the Late Permian (i.e., Changhsingian). Cherts of the Dalong Formation reflect high primary productivity, similar to that of modern-day upwelling zones, which increased the amount of organic matter. These results, combined with previous work, suggest that the long-term oxygen-poor conditions during deposition of the Dalong Formation were caused by restricted water circulation in a unique paleogeographic environment characterized by low latitude and submarine paleo-highlands. Such settings of low latitude characterized by tropical climate favored the growth of siliceous zooplankton in the zones with submarine paleo-highlands having created topographic barriers to water circulation. Restricted bottom-water flow and oceanic anoxia during the Late Permian were likely the main controls on the accumulation of organic-rich rocks. Elevated nutrient levels caused by frequent oceanic upwelling along the western margin of the South China Craton and global volcanism at this time facilitated siliceous organism productivity.

Simultaneous Measurements of Tension and Free H2S in Mesenteric Arteries.

Hydrogen sulfide (H2S), in addition to nitric oxide and carbon monoxide, is the third gasotransmitter and known to cause relaxation in peripheral arteries. Here we describe a method that allows simultaneous measurement of contractility in arteries mounted in an isometric wire myograph and the concentration of free H2S in the lumen of the artery as well as in the organ bath. This method can be used to directly correlate how much free H2S is needed to cause relaxation, which previously has been difficult to answer as H2S can be found in many different forms.

Tissue-dependent variation of hydrogen sulfide homeostasis in anoxic freshwater turtles.

Hydrogen sulfide (H2S) controls numerous physiological responses. To understand its proposed role in metabolic suppression, we measured free H2S and bound sulfane sulfur (BSS) in tissues of the freshwater turtle Trachemys scriptaelegans, a species undergoing strong metabolic suppression when cold and anoxic. In warm normoxic turtles, free H2S was higher in red blood cells (RBCs) and kidney (∼9-10 µmoll-1) than in brain, liver and lung (∼1-2 µmoll-1). These values overall aligned with the tissue H2S-generating enzymatic activity. BSS levels were similar in all tissues (∼0.5 µmoll-1) but ∼100-fold higher in RBCs, which have a high thiol content, suggesting that RBCs function as a circulating H2S reservoir. Cold acclimation caused significant changes in free and bound H2S in liver, brain and RBCs, but anoxia had no further effect, except in the brain. These results show tissue-dependent sulfide signaling with a potential role in brain metabolic suppression during anoxia in turtles.

Methylene Blue Administration During and After Life-Threatening Intoxication by Hydrogen Sulfide: Efficacy Studies in Adult Sheep and Mechanisms of Action.

Exposure to toxic levels of hydrogen sulfide (H2S) produces an acute cardiac depression that can be rapidly fatal. We sought to characterize the time course of the cardiac effects produced by the toxicity of H2S in sheep, a human sized mammal, and to describe the in vivo and in vitro antidotal properties of methylene blue (MB), which has shown efficacy in sulfide intoxicated rats. Infusing NaHS (720 mg) in anesthetized adult sheep produced a rapid dilation of the left ventricular with a decrease in contractility, which was lethal within about 10 min by pulseless electrical activity. MB (7 mg/kg), administered during sulfide exposure, maintained cardiac contractility and allowed all of the treated animals to recover. At a dose of 350 mg NaHS, we were able to produce an intoxication, which led to a persistent decrease in ventricular function for at least 1 h in nontreated animals. Administration of MB, 3 or 30 min after the end of exposure, whereas all free H2S had already vanished, restored cardiac contractility and the pyruvate/lactate (P/L) ratio. We found that MB exerts its antidotal effects through at least 4 different mechanisms: (1) a direct oxidation of free sulfide; (2) an increase in the pool of "trapped" H2S in red cells; (3) a restoration of the mitochondrial substrate-level phosphorylation; and (4) a rescue of the mitochondrial electron chain. In conclusion, H2S intoxication produces acute and long persisting alteration in cardiac function in large mammals even after all free H2S has vanished. MB exerts its antidotal effects against life-threatening sulfide intoxication via multifarious properties, some of them unrelated to any direct interaction with free H2S.

Hyperhomocysteinemia potentiates diabetes-impaired EDHF-induced vascular relaxation: Role of insufficient hydrogen sulfide

Insufficient hydrogen sulfide (H2S) has been implicated in Type 2 diabetic mellitus (T2DM) and hyperhomocysteinemia (HHcy)-related cardiovascular complications. We investigated the role of H2S in T2DM and HHcy-induced endothelial dysfunction in small mesenteric artery (SMA) of db/db mice fed a high methionine (HM) diet. HM diet (8 weeks) induced HHcy in both T2DM db/db mice and non-diabetic db/+ mice (total plasma Hcy: 48.4 and 31.3 µM, respectively), and aggravated the impaired endothelium-derived hyperpolarization factor (EDHF)-induced endothelium-dependent relaxation to acetylcholine (ACh), determined by the presence of eNOS inhibitor N(ω)-nitro-L-arginine methyl ester (L-NAME) and prostacyclin (PGI2) inhibitor indomethacin (INDO), in SMA from db/db mice but not that from db/+ mice. A non-selective Ca2+-active potassium channel (KCa) opener NS309 rescued T2DM/HHcy-impaired EDHF-mediated vascular relaxation to ACh. EDHF-induced relaxation to ACh was inhibited by a non-selective KCa blocker TEA and intermediate-conductance KCa blocker (IKCa) Tram-34, but not by small-conductance KCa (SKCa) blocker Apamin. HHcy potentiated the reduction of free sulfide, H2S and cystathionine γ-lyase protein, which converts L-cysteine to H2S, in SMA of db/db mice. Importantly, a stable H2S donor DATS diminished the enhanced O2- production in SMAs and lung endothelial cells of T2DM/HHcy mice. Antioxidant PEG-SOD and DATS improved T2DM/HHcy impaired relaxation to ACh. Moreover, HHcy increased hyperglycemia-induced IKCa tyrosine nitration in human micro-vascular endothelial cells. EDHF-induced vascular relaxation to L-cysteine was not altered, whereas such relaxation to NaHS was potentiated by HHcy in SMA of db/db mice which was abolished by ATP-sensitive potassium channel blocker Glycolamide but not by KCa blockers. ConclusionsIntermediate HHcy potentiated H2S reduction via CSE-downregulation in microvasculature of T2DM mice. H2S is justified as an EDHF. Insufficient H2S impaired EDHF-induced vascular relaxation via oxidative stress and IKCa inactivation in T2DM/HHcy mice. H2S therapy may be beneficial for prevention and treatment of micro-vascular complications in patients with T2DM and HHcy.

A simple and facile paper-based colorimetric assay for detection of free hydrogen sulfide in prostate cancer cells

In this study, we introduce a new paper-based colorimetric assay fabricated with the 96-well microplate format for cell culture for sensing H2S gas in live cancer cells. The H2S sensing zones on the paper were coated with a polyvinylpyrrolidone (PVP) membrane containing silver/Nafion. The principle of colorimetric detection for free H2S gas is reaction of silver in the coating zones with sulfide to form Ag2S, which is brown in color. First, we optimized the composition of the coating membrane on the paper, including the mixing ratio between PVP and Nafion and the AgNO3 concentration. Next, we examined the sensing performance of the paper assay in the form of sensitivity, selectivity reproducibility, and long-term stability using a known concentration of Na2S standard solution in phosphate buffered saline (PBS) at room temperature (RT) and in cell culture media at 37°C in a 5% CO2 incubator. Our paper assay for H2S detection showed good sensitivity (2.51 color intensity/μM Na2S in PBS, R2=0.9967 at RT; 1.15 color intensity/μM Na2S in cell culture media, R2=0.9933 at 37°C in incubator), high selectivity, excellent reproducibility (relative standard deviation 2.4%), and high stability over 1 month, as well as a limit of detection of 1.4μM Na2S in PBS. Finally, we successfully measured the difference in endogenous H2S concentration between live LNCaP and PC-3 cells, which show different expression of H2S producing enzymes, using this paper-based colorimetric assay. The results were concordant with those of conventional methylene blue assay and western blot analysis. In summary, our paper assay using silver/Nafion-containing PVP membranes is a simple, facile, and reliable tool for the detection of H2S without complicated sample processing or labor-intensive bioanalytical methods We expect that this paper assay could be easily applied to study H2S signaling and pathophysiology in cancer biology, as well as cancer diagnosis and therapeutics.

Elusive Chemistry of Hydrogen Sulfide and Mercaptans in Wine.

This paper summarizes, discusses, and complements recent findings about the fate of H2S and methanethiol (MeSH) during wine storage. Analytical assays to determine free volatile sulfur compounds (VSCs) and brine-releasable (BR-) VSCs in combination with accelerated reductive (AR) aging and micro-oxygenation (MOX) assays allow characterizing the different categories of species able to produce H2S and MeSH and the processes of interconversion. Each wine seems to contain a specific total amount of H2S and MeSH distributed into free, metal-complexed, and oxidized forms (di and polysulfides) interconnected through reversible redox equilibria whose external expression is wine redox potential. Oxidation transforms all mercaptans likely into nonvolatile disulfides and hydrodisulfides. In anoxia, these molecules are spontaneously and quantitatively reduced back. The concomitant accumulation of major wine thiols would provoke complex dissociation and the release of free H2S and MeSH. Additionally, total amounts can increase due to the metal-catalyzed desulfhydration of cysteine and methionine.

Role of thiosulfate in hydrogen sulfide-dependent redox signaling in endothelial cells.

Recent reports have revealed that hydrogen sulfide (H2S) exerts critical actions to promote cardiovascular homeostasis and health. Thiosulfate is one of the products formed during oxidative H2S metabolism, and thiosulfate has been used extensively and safely to treat calcific uremic arteriopathy in dialysis patients. Yet despite its significance, fundamental questions regarding how thiosulfate and H2S interact during redox signaling remain unanswered. In the present study, we examined the effect of exogenous thiosulfate on hypoxia-induced H2S metabolite bioavailability in human umbilical vein endothelial cells (HUVECs). Under hypoxic conditions, we observed a decrease of GSH and GSSG levels in HUVECs at 0.5 and 4 h as well as decreased free H2S and acid-labile sulfide and increased bound sulfide at all time points. Treatment with exogenous thiosulfate significantly decreased the ratio of GSH/GSSG to total sulfide of HUVECs under 0.5 h of hypoxia but significantly increased this ratio in HUVECs under 4 h of hypoxia. These responses reveal that thiosulfate has different effects at low and high doses and under different O2 tensions. In addition, treatment with thiosulfate also diminished VEGF-induced cystathionine-γ-lyase expression and reduced VEGF-induced HUVEC proliferation under both normoxic and hypoxic conditions. These results indicate that thiosulfate can modulate H2S metabolites and signaling under various culture conditions that impact angiogenic activity. Thus, thiosulfate may serve as a unique sulfide donor to modulate endothelial responses under pathophysiological conditions involving angiogenesis.NEW & NOTEWORTHY This report provides new evidence that different levels of exogenous thiosulfate dynamically change discrete sulfide biochemical metabolite bioavailability in endothelial cells under normoxia or hypoxia, acting in a slow manner to modulate sulfide metabolites. Moreover, our findings also reveal that thiosulfate surprisingly inhibits VEGF-dependent endothelial cell proliferation associated with a reduction in cystathionine-γ-lyase protein levels.

The effects of copper fining on the wine content in sulfur off-odors and on their evolution during accelerated anoxic storage

Three different red wines with reductive character have been treated with two different doses of copper sulfate (0.06 and 0.5mg/L) and with a commercial copper-containing product at the recommended dose (0.6mg/L). Wines were in contact with copper one week, centrifuged and stored at 50°C in strict anoxia for 2weeks (up to 7 in one case). Brine-releasable (BR-) and free fractions of Volatile Sulfur Compounds were determined throughout the process. Relevant increases of BR-H2S suggest that those wines contained other H2S precursors non-detectable by the brine dilution method. Copper treatments had two major effects: 1) immediate decrease the levels of free H2S and methanethiol (MeSH); 2) slow the rate at which free H2S (not MeSH) increases during anoxic storage. After 7weeks of anoxia levels of free H2S and MeSH were high and similar regardless of the copper treatment. Higher copper doses could induce the accumulation of BR-H2S.

Highly sensitive potentiometric measuring method for measurement of free H2S in physiologic samples

Hydrogen sulfide in high concentration is a well-known toxic gaseous molecule; nevertheless, H2S is synthetized in mammalls’ body in small concentration participating important physiological processes. It also is implicated in inflammation as a mediator molecule. In this work, a potentiometric air gap cell has been developed in order to apply it in the quantitative study of the H2S accompanied the inflammatory disease in animal experiments. The cell was tested using aqueous standard sample solutions and super-Nernstian response has been obtained in the concentration range of interest and submicromolar lower detection limit, respectively. Animal experiments have been carried out in healthy and inflamed paws of sacrificed mice. The measurements showed significant difference in the H2S level of the control and carrageenan treated tissues of the experimental animals.

H2S concentrations in the heart after acute H2S administration: methodological and physiological considerations.

In this study, we have tried to characterize the limits of the approach typically used to determine H2S concentrations in the heart based on the amount of H2S evaporating from heart homogenates-spontaneously, after reaction with a strong reducing agent, or in a very acidic solution. Heart homogenates were prepared from male rats in control conditions or after H2S infusion induced a transient cardiogenic shock (CS) or cardiac asystole (CA). Using a method of determination of gaseous H2S with a detection limit of 0.2 nmol, we found that the process of homogenization could lead to a total disappearance of free H2S unless performed in alkaline conditions. Yet, after restoration of neutral pH, free H2S concentration from samples processed in alkaline and nonalkaline milieus were similar and averaged ∼0.2-0.4 nmol/g in both control and CS homogenate hearts and up to 100 nmol/g in the CA group. No additional H2S was released from control, CS, or CA hearts by using the reducing agent tris(2-carboxyethyl)phosphine or a strong acidic solution (pH < 2) to "free" H2S from combined pools. Of note, the reducing agent DTT produced a significant sulfide artifact and was not used. These data suggest that 1) free H2S found in heart homogenates is not a reflection of H2S present in a "living" heart and 2) the pool of combined sulfides, released in a strong reducing or acidic milieu, does not increase in the heart in a measurable manner even after toxic exposure to sulfide.

Copper-Complexed Hydrogen Sulfide in Wine: Measurement by Gas Detection Tubes and Comparison of Release Approaches

Recent work suggests that copper complexes may serve as a latent source of free hydrogen sulfide (H2S) and other malodorous volatile thiols during wine storage. However, measurements of these complexes require analytical tools that are unavailable to most wineries. To facilitate additional studies of these compounds, we developed an inexpensive and convenient method for detecting copper-complexed H2S in wine with commercially available colorimetric gas detection tubes. Using brine dilution as a pretreatment, we observed that this approach showed acceptable detection limits (0.34 μg/L) and excellent H2S recovery from both model and real wines. Alternative approaches to H2S release from copper sulfide complexes were also investigated, including ethylenediaminetetraacetic acid (EDTA), neocuproine, ascorbic acid, and tris(2-carboxyethyl)phosphine (TCEP), alone and in combination. EDTA resulted in loss of free H2S. Partial recovery (35 to 70%) of H2S from model wines was achieved with the other reagents, with the release induced by ascorbic acid being of particular interest to winemakers. In a survey of seven commercial wines, the fraction of complexed H2S released by the brine was 80 to 95% of the total H2S pool, with lower releases observed for neocuproine and ascorbic acid. However, in several wines, TCEP treatment released higher concentrations of H2S from unknown precursors.

Hydrogen sulfide metabolism regulates endothelial solute barrier function

Hydrogen sulfide (H2S) is an important gaseous signaling molecule in the cardiovascular system. In addition to free H2S, H2S can be oxidized to polysulfide which can be biologically active. Since the impact of H2S on endothelial solute barrier function is not known, we sought to determine whether H2S and its various metabolites affect endothelial permeability. In vitro permeability was evaluated using albumin flux and transendothelial electrical resistance. Different H2S donors were used to examine the effects of exogenous H2S. To evaluate the role of endogenous H2S, mouse aortic endothelial cells (MAECs) were isolated from wild type mice and mice lacking cystathionine γ-lyase (CSE), a predominant source of H2S in endothelial cells. In vivo permeability was evaluated using the Miles assay. We observed that polysulfide donors induced rapid albumin flux across endothelium. Comparatively, free sulfide donors increased permeability only with higher concentrations and at later time points. Increased solute permeability was associated with disruption of endothelial junction proteins claudin 5 and VE-cadherin, along with enhanced actin stress fiber formation. Importantly, sulfide donors that increase permeability elicited a preferential increase in polysulfide levels within endothelium. Similarly, CSE deficient MAECs showed enhanced solute barrier function along with reduced endogenous bound sulfane sulfur. CSE siRNA knockdown also enhanced endothelial junction structures with increased claudin 5 protein expression. In vivo, CSE genetic deficiency significantly blunted VEGF induced hyperpermeability revealing an important role of the enzyme for barrier function. In summary, endothelial solute permeability is critically regulated via exogenous and endogenous sulfide bioavailability with a prominent role of polysulfides.

Formation and Release of H2S, Methanethiol, and Dimethylsulfide during the Anoxic Storage of Wines at Room Temperature.

A total of 21 different wines (13 reds, 5 whites, and 3 rosés) were kept at 25 °C in anoxia for 379 days. Free and total forms of H2S and methanethiol (MeSH) and dimethylsulfide (DMS) were measured initially and after 117, 221, and 379 days of storage. Levels of free H2S, free and total MeSH, and DMS continuously increased during storage, while levels of total H2S remained essentially unchanged. Average increases of free H2S amount to 6.2 μg/L (from 1.1 to 12.9 μg/L), those of free MeSH to 1.6 μg/L (from 0.7 to 3.5 μg/L), and those of total MeSH to 1.9 μg/L (from 1.1 to 3.0 μg/L), whereas those of DMS were 27.8 μg/L (from 11 to 69 μg/L). The fraction of H2S under free forms significantly increased, suggesting that release is a major factor explaining H2S increases. All increases can be satisfactorily predicted from data obtained at 50 °C.

Adsorption behaviors of hydrogen sulphide on Au(110) nanoslit array surfaces using molecular dynamics simulations

The adsorption behaviour of gas molecules on detector surfaces has a profound influence on the sensitivity of the detector. For this reason, this study used molecular dynamics simulation to explore the dynamic adsorption behaviour of hydrogen sulphide (H2S) molecules on various types of Au surfaces, including a planar Au(1 1 0) structure and three types of slit array structures. The influence of system temperature, adsorbate concentration and the slit width of nanoarrays on diffusivity, average adsorption energy and static adsorption amount were systematically examined. Simulation results indicate that the self-diffusivity of the adsorbate molecules increases with temperature but decreases with adsorbate concentration. At low concentrations (~3 mol/L), each type of Au(1 1 0) surface structure shows good capacity to adsorb all H2S molecules. With increasing concentration at 6.5 mol/L, the high concentration leads to adsorption saturation and many free H2S molecules in the planar Au(1 1 0) structure. Moreover, desorption also begins to appear on the planar structures at a temperature of 300 K (at 6.5 mol/L). The simulation results indicate that the columnar array structures with a slit width ≥5.76 Å allow molecules to swiftly spread into the slits and provide more stable adsorption sites (i.e. with a higher adsorption energy), which can effectively address the issues of high-temperature desorption and adsorption saturation. Particularly at low temperatures (≤100 K), slit structures presented a level of static adsorption of H2S that was 30% to 35 higher than that of planar structures.

Does Protein Sulfuration Take Place in the Heart?

Endogenous hydrogen sulfide (H2S) has been suggested to play a physiological role through protein sulfuration (also referred as sulfhydration) in the regulation of cardiac contractility. The sulfuration is a process by which H2S combines to free cysteine residue on proteins to create persulfide (protein‐S‐SH), changing protein conformation, such as in calcium channels, and functions. Since any new molecule of free H2S produced will immediately combine with the iron present on myoglobin or will be oxidized, our objective was to determine whether increasing endogenous H2S production can actually increase the pool of H2S combined with cysteine residues in the heart.To achieve this goal, we infused exogenous H2S up to a level that will alter cardiac contractility in order to create conditions wherein soluble H2S concentrations increase to levels higher than those reached by endogenous sulfide in the heart. The presence of sulfurated protein in the heart was identified by measuring gaseous H2S released after reducing disulfide bonds. TCEP was chosen as a potent reducing agent instead of dithiothreitol (DTT), previously used for this purpose in the literature, as DTT possesses two thiol groups (−SH) a significant source of H2S. Hearts were removed from control animals and animals exposed to 0.8 mg/min NaHS until cardiac depression‐induced hypotension was produced. The hearts were immediately homogenized in glycine‐NaOH buffer (pH 9), to prevent any evaporation of gaseous H2S. The homogenate was transferred into the 50‐ml chamber filled with sodium phosphate buffer (final pH ~6). At this pH, HS− will easily convert into gaseous H2S, in keeping with H2S partial pressure, and measured in the chamber headspace. The headspace was continuously flowed with N2, and H2S was directly measured by a voltammetry H2S analyzer before and after TCEP (5 mM).We found that the level of H2S produced by TCEP was similar and very low (below 0.5 nmol/g of protein) in both the control and intoxicated hearts, despite levels of free H2S in the blood above 10 microM in the latter. In contrast, the pool of H2S in albumin solution (10% in saline) incubated with H2S for 10 min, increased as a function of H2S concentrations using 0, 2, and 20 microM NaHS and reaching 10.6 ± 1.1, 11.8 ± 0.7 and 13.3 ± 0.4 nmol/g respectively. The change in H2S remained however extremely modest.Toxic levels of soluble H2S were unable to increase the pool of H2S liberated from the heart in a strong reducing environment. This suggests that free H2S in the heart is immediately trapped by other compounds such metallo‐proteins, e.g. myoglobin, present in the heart or is oxidized in the mitochondria before reacting with proteins, in contrast to a solution of albumin unable to produce these effects. Even in the latter, the pool of H2S produced by an increase in free H2S appears to be extremely small and to require toxic levels of sulfide to be affected.

The Ocean redox state evolution and its controls during the Cambrian Series 1–2: Evidence from Lijiatuo Section, South China

Well-exposed Lijiatuo Section was chosen to explore the temporal evolution and controls of the oceanic redox state, primary productivity and seawater sulfate levels during the Cambrian Series 1–2, South China. This section consists of Xiaoyanxi Formation (Fm.) mudstones and Liuchapo Fm. cherts that deposited in the slope and basin environment. Five oxic-anoxic cycles were identified based on V/Sc, Th/U and the enrichment factors of Mo, U, V, Ni and Cu. The Middle-Upper Liuchapo Fm. and the Middle Xiaoyanxi Fm. were deposited under oxic-suboxic conditions, and the rest of the strata were under anoxic conditions. The Re/Mo ratio demonstrated that the oxic-suboxic conditions in the Middle Xiaoyanxi Fm. were accompanied by transient sulfidic conditions, and the rest of the section was underanoxic and non-sulfidic conditions. All the TOC and the enrichment factors of Ba, Ni, Cu, Zn and Cd demonstrated that both the sinking and burial flux of organic matter (OM) in Liuchapo Fm. were lower than that in the overlying Xiaoyanxi Fm. The highest sinking and burial flux of OM in the Xiaoyanxi Fm. appeared at its lower parts; however, the lowest sinking and burial flux of OM in the Xiaoyanxi Fm. appeared in its middle parts. TOC/TS, TS and the vertical trend of δ34Spy demonstrated that the seawater was dominated by low oceanic sulfate levels, which resulted in the absence of free H2S. The rise of the atmospheric oxygen content may be the principal driver for the associated, transient suboxic-oxic and nearly sulfidic environment in the middle Xiaoyanxi Fm.