H2S Production Rate Research Articles

Cystathionine gamma-lyase deficiency exaggerates diethylnitrosamine-induced liver damage in mice

Cystathionine gamma-lyase (CSE) is a key enzyme in reverse transsulfuration pathway and contributes to the majority of H2S generation in liver tissues via cysteine metabolism. Dysfunction of the CSE/H2S system is linked to both chronic and acute liver damage. This study investigated the regulatory role of CSE deficiency on diethylnitrosamine (DEN)-induced liver damage in mice. A single injection of DEN was administered into 4-week-old male CSE knockout (CSE-KO) mice and wild-type (WT) littermates, and the mice were sacrificed at 28 weeks of age. Compared to age-matched WT mice, CSE-KO mice spontaneously developed steatosis with increased oxidative stress and higher expressions of inflammation and fibrosis-related genes at 28-weeks of age. Following DEN injection, CSE-KO mice experienced more severe liver damage in comparison with the WT group as reflected by elevated levels of lipid accumulation, increased activities of alanine aminotransferase and aspartate aminotransferase, higher oxidative stress and fibrosis development, and increased expressions of inflammation and fibrosis-related genes. No visible tumors were observed in both types of mice with DEN treatment. In addition, the expression levels of the three H2S-generating proteins (CSE, cystathionine beta-synthase, and 3-mercaptopyruvate sulfurtransferase) and the H2S production rate in liver tissues were unaffected by DEN. Taken together, our study demonstrates that CSE provides a significant hepatoprotective effect and deficiency of CSE exaggerates DEN-induced liver damage in mice. Based on these findings, it can be suggested that targeting the CSE/H2S signaling pathway could be a potential therapeutic target for the treatment of liver diseases.

Real-time detection of enzymatically formed hydrogen sulfide by pathogenic variants of cystathionine beta-synthase using hemoglobin I of Lucina pectinata as a biosensor

Classical homocystinuria is a rare disease caused by mutations in cystathionine β-synthase (CBS) gene (OMIM 613381). CBS catalyzes the first step of the transsulfuration pathway that converts homocysteine (Hcy) into cystathionine (Cysta) via a number of co-substrates and mechanisms. Formation of Cysta by condensation of Hcy and cysteine (Cys) produces a molar equivalent of hydrogen sulfide (H2S). H2S plays important roles in cognitive and vascular functions. Clinically, patients with CBS deficiency present with vascular, ocular, neurological and skeletal impairments. Biochemically, CBS deficiency manifests with elevated Hcy and reduced concentration of Cysta in plasma and urine. A number of pathogenic variants of human CBS have been characterized by their residual enzymatic activity, but very few studies have examined H2S production by pathogenic CBS variants, possibly due to technical hurdles in H2S detection and quantification. We describe a method for the real-time, continuous quantification of H2S formed by wild-type and pathogenic variants of human recombinant CBS, as well as by fibroblast extracts from healthy controls and patients diagnosed with CBS deficiency. The method takes advantage of the specificity and high affinity of hemoglobin I of the clam Lucina pectinata toward H2S and is based on UV–visible spectrophotometry. Comparison with the gold-standard, end-point H2S quantification method employing monobromobimane, as well as correlations with CBS enzymatic activity determined by LC-MS/MS showed agreement and correlation, and permitted the direct, time-resolved determination of H2S production rates by purified human recombinant CBS and by CBS present in fibroblast extracts. Rates of H2S production were highest for wild-type CBS, and lower for pathogenic variants. This method enables the examination of structural determinants of CBS that are important for H2S production and its possible relevance to the clinical outcome of patients.

The Antioxidant Properties of Glucosinolates in Cardiac Cells Are Independent of H2S Signaling.

The organic sulfur-containing compounds glucosinolates (GSLs) and the novel gasotransmitter H2S are known to have cardioprotective effects. This study investigated the antioxidant effects and H2S-releasing potential of three GSLs ((3E)-4-(methylsulfanyl)but-3-enyl GSL or glucoraphasatin, 4-hydroxybenzyl GSL or glucosinalbin, and (RS)-6-(methylsulfinyl)hexyl GSL or glucohesperin) in rat cardiac cells. It was found that all three GSLs had no effect on cardiac cell viability but were able to protect against H2O2-induced oxidative stress and cell death. NaHS, a H2S donor, also protected the cells from H2O2-stimulated oxidative stress and cell death. The GSLs alone or mixed with cysteine, N-acetylcysteine, glutathione, H2O2, iron and pyridoxal-5'-phosphate, or mouse liver lysates did not induce H2S release. The addition of GSLs also did not alter endogenous H2S levels in cardiac cells. H2O2 significantly induced cysteine oxidation in the cystathionine gamma-lyase (CSE) protein and inhibited the H2S production rate. In conclusion, this study found that the three tested GSLs protect cardiomyocytes from oxidative stress and cell death but independently of H2S signaling.

A Hybrid Data-Physics Framework for Reservoir Performance Prediction with Application to H2S Production

Summary Model-based reservoir management workflows rely on the ability to generate predictions for large numbers of model and decision scenarios. When suitable simulators or models are not available or cannot be evaluated in a sufficiently short time frame, surrogate modeling techniques can be used instead. In the first part of this paper, we describe extensions of a recently developed open-source framework for creating and training flow network surrogate models, called FlowNet. In particular, we discuss functionality to reproduce historical well rates for wells with arbitrary trajectories, multiple perforated sections, and changing well type or injection phase, as one may encounter in large and complex fields with a long history. Furthermore, we discuss strategies for the placement of additional network nodes in the presence of flow barriers. Despite their flexibility and speed, the applicability of flow network models is limited to phenomena that can be simulated with available numerical simulators. Prediction of poorly understood physics, such as reservoir souring, may require a more data-driven approach. We discuss an extension of the FlowNet framework with a machine learning (ML) proxy for the purpose of generating predictions of H2S production rates. The combined data-physics proxy is trained on historical liquid volume rates, seawater fractions, and H2S production data from a real North Sea oil and gas field, and is then used to generate predictions of H2S production. Several experiments are presented in which the data source, data type, and length of the history are varied. Results indicate that, given a sufficient number of training data, FlowNet is able to produce reliable predictions of conventional oilfield quantities. An experiment performed with the ML proxy suggests that, at least for some production wells, useful predictions of H2S production can be obtained much faster and at much lower computational cost and complexity than would be possible with high-fidelity models. Finally, we discuss some of the current limitations of the approach and options to address them.

Observations of comet C/2020 F3 (NEOWISE) with IRAM telescopes

We present the results of millimetre-wave spectroscopic and continuum observations of the comet C/2020 F3 (NEOWISE) undertaken with the Institut de RadioAstronomie Millimétrique (IRAM) 30-m and the NOrthern Extended Millimeter Array (NOEMA) telescopes on 22, 25–27 July, and 7 August 2020. Production rates of HCN, HNC, CH3OH CS, H2CO, CH3CN, H2S, and CO were determined with upper limits on six other species. The comet shows abundances within the range observed for other comets. The CO abundance is low (3.2% relative to water), while H2S is relatively abundant (1.1% relative to water). The H2CO abundance shows a steep variation with heliocentric distance, possibly related to a distributed production from the dust or macro-molecular source. The CH3OH and H2S production rates show a slower decrease post-perihelion than water. There was no detection of the nucleus point source contribution based on the interferometric map of the continuum (implying a size of r < 4.7 km), but this yielded an estimate of the dust production rate, leading to a relatively low dust-to-gas ratio of 0.7 ± 0.3 on 22.4 July 2020.

Looping upcycling SO2 into value-added H2S by fast-induced reduction process for heavy metals treatment in nonferrous smelting industry

Reversing the pollution to a value-added product can simultaneously alleviate environmental problems and achieve a circular economy. In nonferrous smelting industries, SO2 is hazardous pollution that should be disposed, but H2S is usually required for heavy metals (HMs) treatment. Herein, we developed a field SO2 looping upcycling technology for value-added H2S production to meet the HMs treatment. Results demonstrated that the H2 could accelerate the kinetic rate and decrease the energy barriers of the SO2 and CH4 reaction by a fast-induced reduction process (fast-IRP). Thus, the H2S production rate in fast-IRP was three times higher than the direct reaction between SO2 and CH4. Meanwhile, the T90 temperature (690 °C) was significantly lower than SO2 and CH4 reaction about 90 °C. Fast-IRP also can resist the SO2 poison effect and prevent sulfation, enhancing the activity, long-term stability (no visible efficiency loss in 50 h), and selectivity (nearly 100 %). Integrated the characterizations and DFT results, we found that the H2 would firstly attack the uncoordinated sulfur species with a hydrodesulfurization process to form the HS-/H2S intermediate species with a lower energy barrier (1.57 eV) than the CH4 (2.83 eV), which can facilely protect the Mo-terminated to avoid sulfation poison. Subsequently, the HS-/H2S* species would react with the adsorbed SO2 to Sx* to support the deep-reduction reaction for H2S production. This work proposes an actual SO2 field utilization route and also will enlighten an induced redox cycle in the sulfur reduction over the pre-sulfurized catalysts.

Cloning of cucumber LCD and DES gene and their response to abiotic stress

Hydrogen sulfide (H2S) has been reported as a significant gasotransmitter, which involved in a range of physiological processes in plant, including growth, development and abiotic stresses resistence. Enzymatic activity assays showed that L-cysteine desulfhydrase (L-CD) was primarily responsible for the generation of endogenous H2S in plant. In this work, we identified two activated L-CDs from cucumber 9930 varieties. Firstly, three L-CD genes, CsaLCD, CsaDES1 and CsaDES2 were acquired from tBLASTn algorithm-based search of NCBI and a cucumber genomic database (http://www.icugi.org), using the Arabidopsis LCD and DES1 protein sequences as the query. The CDS of CsaLCD (Csa2P034800.1) and CsaDES2 (Csa1P574810.1) were amplified successfully by PCR, while the CsaDES1 (Csa1P574800.1) could not be obtained in the case of cDNA as the template. The recombinant proteins His-CsaLCD and His-CsaDES2, which were purified from E.coli BL21 strain, exhibited evident L-CD activity. The mRNA level of CsaLCD was induced by 37 °C, low light or PEG8000 treatments, and CsaDES2 was induced by 4 °C or 37 °C treatments, while CsaDES1 itself has no expression. The CDS of CsaLCD or CsaDES2 with 35S promoter were transformated into lcd/des mutant of Arabidopsis, and both their heterologous expression restored the H2S production rate, as well as enhancing the resistance to osmosis stress of the transgenetic seedlings. Therefore, CsaLCD and CsaDES2 are the main enzymes for endogenous H2S production in cucumber, and play a pivotal role in H2S generation during stresses responses.

Prokaryotic expression, purification and functional identification of epidermal pattern factors in Arabidopsis thaliana

Stomatal density is important for crop yield. In this paper, we studied the epidermal pattern factors (EPFs) related to stomatal development. Prokaryotic expression vectors were constructed to obtain EPFs. Then the relationship between EPFs and hydrogen sulfide (H2S) was established. First, AtEPF1, AtEPF2 and AtEPFL9 were cloned and constructed to pET28a vectors. Then recombinant plasmids pET28a-AtEPF1, pET28a-AtEPF2 and pET28a-AtEPFL9 were digested and sequenced, showing successful construction. Finally, they were transformed into E. coli BL21(DE3) separately and induced to express by isopropyl β-D-galactoside (IPTG). The optimized expression conditions including IPTG concentration (0.5, 0.3 and 0.05 mmol/L), temperature (28 °C, 28 °C and 16 °C) and induction time (16 h, 16 h and 20 h) were obtained. The bands of purified proteins were about 18 kDa, 19 kDa and 14.5 kDa, respectively. In order to identify their function, the purified AtEPF2 and AtEPFL9 were presented to Arabidopsis thaliana seedlings. Interestingly, the H2S production rate decreased or increased compared with the control, showing significant differences. That is, EPFs affected the production of endogenous H2S in plants. These results provide a foundation for further study of the relationship between H2S and EPFs on stomatal development, but also a possible way to increase the yield or enhance the stress resistance.

Increased sulfate availability in saline water promotes hydrogen sulfide production in fish organic waste

The risk of hydrogen sulfide (H2S) production can be a challenge in marine land-based recirculating aquaculture systems (RAS). Hydrogen sulfide is a toxic gas that can cause massive fish mortality even at low concentrations, and in addition, serious odour problems in the surroundings. It is a bacterial by-product originating from the degradation of organic matter in sulfur-rich waters such as marine waters. In order to hinder H2S production in marine land-based RAS, more information on the H2S production conditions and the associated microbiology is needed. In this study, the production of H2S from rainbow trout (Oncorhynchus mykiss) organic waste was examined using a novel H2S measurement method under a range of salinities (0, 5, 10, 15, 25 and 35 g/L) in anaerobic mixed reactors, and the microbial communities as well as abundance of sulfate reducing bacteria (SRB) were characterized. The maximum H2S concentration increased from 23.1 ± 8.2 mg H2S/L at 0 g/L salinity to 153.9 ± 34.1 mg H2S/L at 35 g/L salinity. Similarly, the H2S production rate increased from 5.6 ± 0.2 at 0 g/L salinity to 26.4 ± 12.7 mg of H2S produced per day at 35 g/L salinity. The highest H2S production was recorded after increased availability of volatile fatty acids, which were produced by fermentative bacteria from phyla Firmicutes and Bacteroidetes that dominated the microbial communities after day 5. The traditional sulfate reducing bacteria (SRB) were found only at 0 and 5 g/L salinity, while at higher salinities, H2S production was carried out by novel unquantifiable SRB. The results demonstrate that H2S can be a pronounced problem in marine RAS, although it can be controlled through preventing anaerobic conditions within the system.

Prospects for biogas production and H2S control from the anaerobic digestion of cattle manure: The influence of microscale waste iron powder and iron oxide nanoparticles

Improving the quality and quantity of biogas usually requires pre-treatment to maximize methane yields and/or post-treatment to remove H2S, which involves considerable energy consumption and higher costs. Therefore, this study proposes a cost-effective method for the enhanced anaerobic digestion (AD) of dairy manure (DM) without pre/post-treatment by directly adding waste iron powder (WIP) and iron oxide nanoparticles (INPs) to batch digesters. The results showed that the addition of iron in the form of microscale WIP (generated from the laser cutting of iron and steel) at concentrations of 100 mg/L, 500 mg/L, and 1000 mg/L improved methane yields by 36.99%, 39.36%, and 56.89%, respectively. In comparison, the equivalent dosages of INPs improved yields by 19.74%, 18.14%, and 21.11%, respectively. Additionally, the highest WIP dose (1000 mg/L) achieved the maximum improvement in the rate of hydrolysis (k), which was 1.25 times higher than in control reactions, and a maximum biomethane production rate (Rmax) of 0.045 L/gVS/d according to kinetic analysis models (i.e., first-order and the Gompertz kinetic models). The rate of H2S production was also significantly reduced (by 45.20%, 58.16%, and 77.24%) using the three WIP concentrations in comparison with INPs (which achieved reductions of 33.59%, 46.30%, and 53.52%, respectively). Therefore, the direct mixing of WIP with cattle manure is proposed as a practical and economical means of addressing complex and high-cost pre- and post-treatments that are otherwise required in the digestion process.

Propargylglycine decreases neuro-immune interaction inducing pain response in temporomandibular joint inflammation model

The mechanisms underlying temporomandibular disorders following orofacial pain remain unclear. Hydrogen sulfide (H2S), a newly identified gasotransmitter, has been reported to modulate inflammation. Cystathionine γ-lyase (CSE) is responsible for the systemical production of H2S, which exerts both pro- and antinociceptive effects through inflammation. In the current study, we investigated whether the endogenous H2S production pathway contributes to arousal and maintenance of orofacial inflammatory pain, through the investigation of the effects of a CSE inhibitor, propargyglycine (PAG), in a rat CFA (Complete Freund Adjuvant)-induced temporomandibular inflammation model to mimic persistent pain in the orofacial region. For this, rats received either CFA or saline in the temporomandibular joints (TMJs), and after 3 or 14 days, they received a single injection of PAG or saline and were evaluated for nociception with the von Frey and formalin test. Also, pro-inflammatory cytokines, tumor necrosis factor-α (TNF-α), and interleukin-1β (IL-1β) were analyzed in TMJs and trigeminal ganglion (TG). In this last one, glial cells reactivity was also verified. Endogenous H2S production rate were measured in both, TMJ and TG. Our results indicated decreased allodynia and hyperalgesic responses in rats submitted to CFA after injection of PAG. Moreover, PAG inhibited leucocyte migration to temporomandibular synovial fluid after 3 and 14 days of inflammation. PAG was able to reduce levels of CBS, CSE, TNF-α, and IL-1β in the TMJ and TG, after 13 days of CFA injection. The observed increased activation of glial cells in the trigeminal ganglia on the 14th day of inflammation can be prevented by the highest dose of PAG. Finally, CBS and CSE expression, and endogenous H2S production rate in the TMJ and TG was found higher in rats with persistent temporomandibular inflammation compared to rats injected with saline and PAG was able to prevent this elevation. Our results elucidated the molecular mechanisms by which H2S exerts its pro-inflammatory and pro-nociceptive role in the orofacial region by alterations in both local tissue and TG.

The gasotransmitter hydrogen sulfide inhibits transepithelial anion secretion of pregnant mouse endometrial epithelium

Endometrial epithelium exhibits a robust ion transport activity required for dynamical regulation of uterine fluid environment and thus embryo implantation. However, there still lacks a thorough understanding of the ion transport processes and regulatory mechanism in peri-implantation endometrial epithelium. As a gaseous signaling molecule or gasotransmitter, hydrogen sulfide (H2S) regulates a myriad of cellular and physiological processes in various tissues, including the modulation of ion transport proteins in epithelium. This study aimed to investigate the effects of H2S on ion transport across mouse endometrial epithelium and its possible role in embryo implantation. The existence of endogenous H2S in pregnant mouse uterus was tested by the detection of two key H2S-generating enzymes and measurement of H2S production rate in tissue homogenates. Transepithelial ion transport processes were electrophysiologically assessed in Ussing chambers on early pregnant mouse endometrial epithelial layers, demonstrating that H2S suppressed the anion secretion by blocking cystic fibrosis transmembrane conductance regulator (CFTR). H2S increased intracellular Cl− concentration ([Cl−]i) in mouse endometrial epithelial cells, which was abolished by pretreatment with the CFTR selective inhibitor CFTRinh-172. The cAMP level in mouse endometrial epithelial cells was not affected by H2S, indicating that H2S blocked CFTR in a cAMP-independent way. In vivo study showed that interference with H2S synthesis impaired embryo implantation. In conclusion, our study demonstrated that H2S inhibits the transepithelial anion secretion of early pregnant mouse endometrial epithelium via blockade of CFTR, contributing to the preparation for embryo implantation.

Hydrogen Sulfide Regulates Energy Production to Delay Leaf Senescence Induced by Drought Stress in Arabidopsis.

Hydrogen sulfide (H2S) is a novel gasotransmitter in both mammals and plants. H2S plays important roles in various plant developmental processes and stress responses. Leaf senescence is the last developmental stage and is a sequential degradation process that eventually leads to leaf death. A mutation of the H2S-producing enzyme-encoding gene L-cysteine desulfhydrase1 (DES1) leads to premature leaf senescence but the underlying mechanisms are not clear. In this present study, wild-type, DES1 defective mutant (des1) and over-expression (OE-DES1) Arabidopsis plants were used to investigate the underlying mechanism of H2S signaling in energy production and leaf senescence under drought stress. The des1 mutant was more sensitive to drought stress and displayed accelerated leaf senescence, while the leaves of OE-DES1 contained adequate chlorophyll levels, accompanied by significantly increased drought resistance. Under drought stress, the expression levels of ATPβ-1, -2, and -3 were significantly downregulated in des1 and significantly upregulated in OE-DES1, and ATPε showed the opposite trend. Senescence-associated gene (SAG) 12 correlated with age-dependent senescence and participated in the drought resistance of OE-DES1. SAG13, which was induced by environmental factors, responded positively to drought stress in des1 plants, while there was no significant difference in the SAG29 expression between des1 and OE-DES1. Using transmission electron microscopy, the mitochondria of des1 were severely damaged and bubbled in older leaves, while OE-DES1 had complete mitochondrial structures and a homogeneous matrix. Additionally, mitochondria isolated from OE-DES1 increased the H2S production rate, H2S content and ATPase activity level, as well as reduced swelling and lowered the ATP content in contrast with wild-type and des1 significantly. Therefore, at subcellular levels, H2S appeared to determine the ability of mitochondria to regulate energy production and protect against cellular aging, which subsequently delayed leaf senescence under drought-stress conditions in plants.

Prediction of H2S production rate in sewer systems using the Z model: a case study in Dehloran city, Iran

Modeling of H2S buildup in sewers is performed due to the health problems associated with the high concentration of H2S, destruction of non-resistant structures in corrosion processes, and high costs of repairing corroded concrete sewer pipes. This analytical study aimed to predict the risk of H2S production in the sewage collection network of Dehloran city, Iran using the Z model. In total, 11 main sewage lines with various diameters were selected for wastewater sampling. For each pipeline, two samples per month were collected and processed for the analysis of various parameters in order to determine the H2S production based on the Z value. Biochemical oxygen demand, chemical oxygen demand, and SO4-2 values in warm seasons were 117, 291, and 251 mg/l, while they were 101, 247, and 234 mg/l in cold seasons, respectively. In all the samples, the Z value was >13,000. In addition, the Z level was higher in warm seasons (Z value in guaranteed H2S production category) compared to cold seasons (Z value in a large possibility of H2S production category), which could be due to the high temperature and anaerobic decomposition of organic matter in summer. A significant correlation was also observed between the Z value in different seasons and various diameters of the sewers. Considering the high risk of H2S production, it is recommended that proper scheming and planning be performed to eliminate this gas and prevent corrosion.

Effect of O2 enrichment on acid gas oxidation and formation of COS and CS2 in a rich diffusion flame

O2 enriched combustion is applied to a non-premixed acid gas (AG) flame, in which AG and air are injected separately into a vertical reactor via a coaxial burner at atmosphere pressure. Equilibrium predictions of AG oxidation were performed using Aspen Plus and lab-scale experiments with particular focus on formation of COS and CS2 were undertaken under different combustion conditions. The effects of equivalence ratio (Φ=0.8, 1.0 and 3.0) and initial O2 concentration (OC) in air (21, 30 and 50vol%) have been studied and the flame is interpreted by analyzing the axial temperature and species concentration distributions along the reactor.AG diffusion flame could be divided into three zones, namely AG decomposition, oxidation and complex reaction zones, among which decomposition zone is tightly associated with formation of COS and CS2. It is shown that Φ generally determines the flame temperature and controls the H2S oxidation degree and production rate of H2 and CO. Reactivity of CO2 primarily expressed via CO2+H=CO+OH during the fuel rich flame. COS is primarily produced in the flame inner core via reactions jointing CO2 and sulfur species, consequently its formation shows a low sensitivity to Φ in the air-supplied flame. O2 enrichment basically contributes to higher flame temperature, accelerated H2S oxidation and advanced CO production. Also an increasing tendency for H2S to decompose into H2 and S2 can be observed. And this factor directly triggers the formation of COS and CS2 via increasing the presence of CO and sulfur species. COS is formed within extended channels involving the primary production by reaction of SH+CO2 and the secondary by reactions between CO and sulfur species. The CS2 formation is more complex, mainly comprises of reactions involving CS intermediate and evidently it is enhanced under O2 enriched combustion.

Effect of Physical Exercise on the Febrigenic Signaling is Modulated by Preoptic Hydrogen Sulfide Production.

We tested the hypothesis that the neuromodulator hydrogen sulfide (H2S) in the preoptic area (POA) of the hypothalamus modulates the febrigenic signaling differently in sedentary and trained rats. Besides H2S production rate and protein expressions of H2S-related synthases cystathionine β-synthase (CBS), 3-mercaptopyruvate sulfurtransferase (3-MPST) and cystathionine γ-lyase (CSE) in the POA, we also measured deep body temperature (Tb), circulating plasma levels of cytokines and corticosterone in an animal model of systemic inflammation. Rats run on a treadmill before receiving an intraperitoneal injection of lipopolysaccharide (LPS, 100 μg/kg) or saline. The magnitude of changes of Tb during the LPS-induced fever was found to be similar between sedentary and trained rats. In sedentary rats, H2S production was not affected by LPS. Conversely, in trained rats LPS caused a sharp increase in H2S production rate that was accompanied by an increased CBS expression profile, whereas 3-MPST and CSE expressions were kept relatively constant. Sedentary rats showed a significant LPS-induced release of cytokines (IL-1β, IL-6, and TNF-α) which was virtually abolished in the trained animals. Correlation between POA H2S and IL-6 as well as TNF-α was observed. Corticosterone levels were augmented after LPS injection in both groups. We found correlations between H2S and corticosterone, and corticosterone and IL-1β. These data are consistent with the notion that the responses to systemic inflammation are tightly regulated through adjustments in POA H2S production which may play an anti-inflammatory role downmodulating plasma cytokines levels and upregulating corticosterone release.

Involvement of endogenous central hydrogen sulfide (H2S) in hypoxia-induced hypothermia in spontaneously hypertensive rats.

Spontaneously hypertensive rats (SHR) display autonomic imbalance and abnormal body temperature (Tb) adjustments. Hydrogen sulfide (H2S) modulates hypoxia-induced hypothermia, but its role in SHR thermoregulation is unknown. We tested the hypothesis that SHR display peculiar thermoregulatory response to hypoxia and that endogenous H2S overproduced in the caudal nucleus of the solitary tract (NTS) of SHR modulates this response. SHR and Wistar rats were microinjected into the fourth ventricle with aminooxyacetate (AOA, H2S-synthezing enzyme inhibitor) or sodium sulfide (Na2S, H2S donor) and exposed to normoxia (21% inspired O2) or hypoxia (10% inspired O2, 30 min). Tb was continuously measured, and H2S production rate was assessed in caudal NTS homogenates. In both groups, AOA, Na2S, or saline (i.e., control; 1 μL) did not affect euthermia. Hypoxia caused similar decreases in Tb in both groups. AOA presented a longer latency to potentiate hypoxic hypothermia in SHR. Caudal NTS H2S production rate was higher in SHR. We suggest that increased bioavailability of H2S in the caudal NTS of SHR enables the adequate modulation of excitability of peripheral chemoreceptor-activated NTS neurons that ultimately induce suppression of brown adipose tissue thermogenesis, thus accounting for the normal hypoxic hypothermia.

Vitamin B6 nutritional status and cellular availability of pyridoxal 5'-phosphate govern the function of the transsulfuration pathway's canonical reactions and hydrogen sulfide production via side reactions.

The transsulfuration pathway (TS) acts in sulfur amino acid metabolism by contributing to the regulation of cellular homocysteine, cysteine production, and the generation of H2S for signaling functions. Regulation of TS pathway kinetics involves stimulation of cystathionine β-synthase (CBS) by S-adenosylmethionine (SAM) and oxidants such as H2O2, and by Michaelis-Menten principles whereby substrate concentrations affect reaction rates. Although pyridoxal phosphate (PLP) serves as coenzyme for both CBS and cystathionine γ-lyase (CSE), CSE exhibits much greater loss of activity than CBS during PLP insufficiency. Thus, cellular and plasma cystathionine concentrations increase in vitamin B6 deficiency mainly due to the bottleneck caused by reduced CSE activity. Because of the increase in cystathionine, the canonical production of cysteine (homocysteine→cystathionine→cysteine) is largely maintained even during vitamin B6 deficiency. Typical whole body transsulfuration flux in humans is 3-7μmol/h per kg body weight. The invivo kinetics of H2S production via side reactions of CBS and CSE in humans are unknown but they have been reported for cultured HepG2 cells. In these studies, cells exhibit a pronounced reduction in H2S production capacity and rates of lanthionine and homolanthionine synthesis in deficiency. In humans, plasma concentrations of lanthionine and homolanthionine exhibit little or no mean change due to 4-wk vitamin B6 restriction, nor do they respond to pyridoxine supplementation of subjects in chronically low-vitamin B6 status. Wide individual variation in responses of the H2S biomarkers to such perturbations of human vitamin B6 status suggests that the resulting modulation of H2S production may have physiological consequences in a subset of people. Supported by NIH grant DK072398. This paper refers to data from studies registered at clinicaltrials.gov as NCT01128244 and NCT00877812.

WRKY transcription factors down-regulate the expression of H2S-generating genes, LCD and DES in Arabidopsis thaliana

Hydrogen sulfide (H2S) is a gasotransmitter playing a vital role in response to biotic and abiotic stress for plants. In order to understand the transcriptional regulation of the genes that are responsible for endogenous H2S generation, the promoter sequences of L-cysteine desulfhydrase (LCD), D-cysteine desulfhydrase (DCD1, DCD2), desulfhydrase (DES) and nitrogen fixation synthetase (NFS1, NFS2) were analyzed. They are all found to contain a W-box, a characteristic core binding site for the plant WRKY transcription factors, which have important roles in the plant’s responses to numerous stresses by modifying the expression patterns of their target genes. An electrophoretic mobility shift assay indicated that WRKY18 and WRKY60 interacted with the W-box in the promoters of the LCD, DCD1, DCD2, DES and NFS2 genes, whereas WRKY40 bound to the W-box of the NFS1 promoter. The expression levels of the LCD, DES and DCD1 genes were up-regulated, but the DCD2 was down-regulated in the plants with WRKY18, WRKY40 or WRKY60 mutations. The plants with double and triple mutations of WRKY18, WRKY40 and WRKY60 had a higher rate of H2S production during cadmium stress and were more resistant to the cadmium stress than the wild type or single mutants. These results suggest that WRKY transcription factors regulate the H2S signaling pathway in plants, allowing them to cope with cadmium stress.

Inhibitory effect of hydrogen sulfide on ozone-induced airway inflammation, oxidative stress, and bronchial hyperresponsiveness.

Exposure to ozone has been associated with airway inflammation, oxidative stress, and bronchial hyperresponsiveness. The goal of this study was to examine whether these adverse effects of ozone could be prevented or reversed by hydrogen sulfide (H2S) as a reducing agent. The H2S donor sodium (NaHS) (2 mg/kg) or vehicle (PBS) was intraperitoneally injected into mice 1 hour before and after 3-hour ozone (2.5 ppm) or air exposure, and the mice were studied 24 hours later. Preventive and therapeutic treatment with NaHS reduced the ozone-induced increases in the total cells, including neutrophils and macrophages; this treatment also reduced levels of cytokines, including TNF-α, chemokine (C-X-C motif) ligand 1, IL-6, and IL-1β levels in bronchial alveolar lavage fluid; inhibited bronchial hyperresponsiveness; and attenuated ozone-induced increases in total malondialdehyde in bronchoalveolar lavage fluid and decreases in the ratio of reduced glutathione/oxidized glutathione in the lung. Ozone exposure led to decreases in the H2S production rate and in mRNA and protein levels of cystathionine-β-synthetase and cystathionine-γ-lyase in the lung. These effects were prevented and reversed by NaHS treatment. Furthermore, NaHS prevented and reversed the phosphorylation of p38 mitogen-activated protein kinase and heat shock protein 27. H2S may have preventive and therapeutic value in the treatment of airway diseases that have an oxidative stress basis.