Small, gaseous molecules such as nitric oxide, carbon monoxide and hydrogen sulfide are produced as signalling molecules in mammalian cells. Here, we show that low concentrations of cyanide are generated endogenously in various mammalian tissues and cells. We detect cyanide in several cellular compartments of human cells and in various tissues and the blood of mice. Cyanide production is stimulated by glycine, occurs at the low pH of lysosomes and requires peroxidase activity. When generated at a specific rate, cyanide exerts stimulatory effects on mitochondrial bioenergetics, cell metabolism and cell proliferation, but impairs cellular bioenergetics at high concentrations. Cyanide can modify cysteine residues via protein S-cyanylation, which is detectable basally in cells and mice, and increases in response to glycine. Low-dose cyanide supplementation exhibits cytoprotective effects in hypoxia and reoxygenation models in vitro and in vivo. Conversely, pathologically elevated cyanide production in nonketotic hyperglycinaemia is detrimental to cells. Our findings indicate that cyanide should be considered part of the same group of endogenous mammalian regulatory gasotransmitters as nitric oxide, carbon monoxide and hydrogen sulfide.

Hydrogen sulfide (H2S) gas therapygarners significant attention for its potential to improve outcomes in various disease treatments. The quantitative control of H2S release is crucial for effective the rapeutic interventions; however, traditional researchon H2S therapy frequently utilizes static release models and neglects the dynamic nature of blood flow. In this study, we propose a novel slow-release in-situ H2S release model that leverages the dynamic hydrolysis of H2S donorswithin the bloodstream. Calcium sulfide nanoparticles (CaS NPs) withmicrosolubility characteristics exhibit continuous H2S release, surpassing 24 h at normal blood flow velocities. The extended-release profile demonstrates superior potential in aligning with the bell-shapedpharmacological effect of H2S, compared to NaHS. Moreover, we synthesisedrare earth-doped CaS NPs (CaS: Eu2+, Sm3+ NPs) tha texhibit persistent luminescence, enabling visualisation of the continuous H2S release in trials. Our results demonstrate that lowdose CaS: Eu2+, Sm3+ NPs significantly reduces seizureduration to 1.2 ± 0.7 minutes, while high dose effectively suppresses colontumor growth with a tumor inhibition rate of 54%. Remarkably, these findings closely resemble endogenous H2S levels in treating epilepsy and tumors. This innovative slow-release, in-situ H2S the rapeutic approach via hydrolysis rejuvenates the development of H2S-basedtherapeutics.

In this study, we investigated the interaction between the H2S ligand and the heme pocket of hemoglobin I (HbI) of Lucina pectinata for the wild-type protein; three known mutations where distal glutamine is replaced by hydrophobic valine (Gln64Val) and hydrophilic histidine in both protonation forms (Gln64Hisϵ and Gln64Hisδ); five known mutations of the so-called phenyl cage, replacing the hydrophobic phenylalanines Phe29 and Phe43 with tyrosine (Tyr), valine (Val), or leucine (Leu); and two additional mutations, Phe68Tyr and Phe68Val, in order to complement previous studies with new insights about the binding mechanism at the molecular level. A particular focus was on the intrinsic strengths of the chemical bonds involved, utilizing local vibrational force constants based on combined quantum mechanical-molecular mechanical calculations. Wild-type protein and mutations clustered into two distinct groups: Group 1 protein systems with a proton acceptor in the distal protein pocket, close to one of the H2S bonds, and Group 2 protein systems without a hydrogen acceptor close by in the active site of the protein. According to our results, the interactions between H2S and HbI of Lucina pectinata involve two important elements, namely, binding of H2S to Fe of the heme group, followed by the proton transfer from the HS bond to the distal residue. The distal residue is additionally stabilized by a second proton transfer from the distal residue to COO- of the propionate group in heme. We could identify the FeS bond as a key player and discovered that the strength of this bond depends on two mutual factors, namely, the strength of the HS bond involved in the proton transfer and the electrostatic field of the protein pocket qualifying the FeS bond as a sensitive probe for monitoring changes in H2S ligation upon protein mutations. We hope our study will inspire and guide future experimental studies, targeting new promising mutations such as Phe68Tyr, Phe68Val, or Phe43Tyr/Phe68Val.

Reactive oxygen species (ROS) and reactive sulfur species (RSS) are involved in many physiological processes and act as collaborators with crosstalk. As an important member of gasotransmitters and RSS, hydrogen sulfide (H₂S) carries out signaling functions at submicromolar levels because of its high reactivity. Mechanisms of dynamic regulation of ROS and H₂S production are poorly understood, and the development of a highly selective and organelle-targeted chemical tool will advance the further understanding of H₂S chemical biology and ROS/RSS crosstalk. Herein, we report a highly selective and sensitive, endoplasmic reticulum (ER)-targeted fluorescent probe (ER-BODIPY-NBD) for revealing cisplatin-induced H₂S biogenesis for the first time. The probe demonstrates a 152-fold fluorescence enhancement at 520 nm after reaction with H₂S to release a bright BODIPY product (quantum yield 0.36). The probe is highly selective toward H₂S over biothiols, ER-targeted, and biocompatible. In addition, the probe was successfully employed to track H₂S biogenesis in live cells via stimulation from exogenous hypochlorous acid and the drug cisplatin.

An unprecedented H₂S release from cysteine esters and amides (CysO/NHR) under physiological conditions was discovered and the plausible mechanism was proposed. Alkylation of the amino moiety of cysteine esters enables the H₂S release to be tuned and further provides support to the mechanistic insights. This discovery not only provides new insights into several fundamental science issues including non-enzymatic H₂S-produced pathways, but also inspires new tunable cleavable motifs for sustained release of arylthiols and even for prodrug design.

Following the discovery of nitric oxide (NO) and carbon monoxide (CO), hydrogen sulfide (H₂ S) has been identified as the third gasotransmitter in humans. Increasing evidence have shown that H₂ S is of preventive or therapeutic effects on diverse pathological complications. As a consequence, it is of great significance to develop suitable approaches of H₂ S-based therapeutics for biomedical applications. H₂ S-releasing agents (H₂ S donors) play important roles in exploring and understanding the physiological functions of H₂ S. More importantly, accumulating studies have validated the theranostic potential of H₂ S donors in extensive repertoires of in vitro and in vivo disease models. Thus, it is imperative to summarize and update the literatures in this field. In this review, first, the background of H₂ S on its chemical and biological aspects is concisely introduced. Second, the studies regarding the H₂ S-releasing compounds are categorized and described, and accordingly, their H₂ S-donating mechanisms, biological applications, and therapeutic values are also comprehensively delineated and discussed. Necessary comparisons between related H₂ S donors are presented, and the drawbacks of many typical H₂ S donors are analyzed and revealed. Finally, several critical challenges encountered in the development of multifunctional H₂ S donors are discussed, and the direction of their future development as well as their biomedical applications is proposed. We expect that this review will reach extensive audiences across multiple disciplines and promote the innovation of H₂ S biomedicine.

Symbiosis is found throughout nature, but perhaps nowhere is it more fundamental than mitochondria in all eukaryotes. Since mitochondria were discovered and mechanisms of oxygen reduction characterized, an understanding gradually emerged that these organelles were involved not just in the combustion of oxygen, but also in the sensing of oxygen. While multiple hypotheses exist to explain the mitochondrial involvement in oxygen sensing, key elements are developing that include potassium channels and reactive oxygen species. To understand how mitochondria contribute to oxygen sensing, it is informative to study a model system which is naturally adapted to survive extended periods without oxygen. Amongst air-breathing vertebrates, the most highly adapted are western painted turtles (Chrysemys picta bellii), which overwinter in ice-covered and anoxic water bodies. Through research of this animal, it was postulated that metabolic rate depression is key to anoxic survival and that mitochondrial regulation is a key aspect. When faced with anoxia, excitatory neurotransmitter receptors in turtle brain are inhibited through mitochondrial calcium release, termed "channel arrest". Simultaneously, inhibitory GABAergic signalling contributes to the "synaptic arrest" of excitatory action potential firing through a pathway dependent on mitochondrial depression of ROS generation. While many pathways are implicated in mitochondrial oxygen sensing in turtles, such as those of adenosine, ATP turnover, and gaseous transmitters, an apparent point of intersection is the mitochondria. In this review we will explore how an organelle that was critical for organismal complexity in an oxygenated world has also become a potentially important oxygen sensor.

The thiolysis of NBD piperazinyl amine (NBD-PZ) is highly selective for H₂S over GSH and has been widely used for the development of many H₂S fluorescent probes. Whether the NBD amine in H₂S-specific probes could be a fluorescent quencher should be further clarified, because NBD amines have been used as environment-sensitive fluorophores for many years. Here, we compared the properties of NBD-based secondary and tertiary amines under the same conditions. For example, the emission of NBD-N(Et)2 is much smaller in water and less responsive to changes in polarity than that of NBD-NHEt. The emission of NBD-PZ-TPP is also smaller than that of NBD-NH-TPP both in aqueous buffer and in live cells. In addition, confocal bioimaging signals of NBD-PZ-TPP with excitation at 405 nm and 454 nm are much weaker than that at 488 nm. Based on these results as well as the previous work on NBD-based probes, we discuss and summarize the design strategies and sensing mechanisms for different NBD-based H₂S probes. Moreover, NBD-PZ-TPP may be a useful tool for reaction with and imaging of mitochondrial H₂S in live cells. This work should be useful for clarification of the roles of NBD in H₂S-specific fluorescent probes as well as for facilitating the development of future NBD-based probes.

Hydrogen sulfide (H₂S), previously recognized as a toxic gas, has emerged as an important gaseous signaling molecule along with nitric oxide, carbon monoxide and also hydrogen. H₂S can be endogenously produced in the mammalian body at a very low level for various pathophysiological processes. Notably, H₂S can interact with several essential metals in the body such as iron, copper, nickel, and zinc to carry out specific functions. The interactions of H₂S with metal-binding proteins have been shown to aid in its signal transduction and cellular metabolism. In addition, H₂S is capable of providing a cytoprotective role against metal toxicity. As the research in the field of H₂S signaling in biology and medicine increases, much progresses have been developed for detecting H₂S via interaction with metals. In this review, the interaction of H₂S with metals, specifically in regard to metal-driven metabolism of H₂S, the protection against metal toxicity by H₂S and the detection of H₂S using metals will be discussed. Discovering the interactions of this gasotransmitter with metals is important for determining the mechanisms underlying the cellular functions of H₂S as well as developing novel therapeutic avenues.

H₂S is a gaseous signaling molecule that is involved in many physiological and pathological processes. In general, the level of intracellular H₂S (<1 μM) is much lower than that of GSH (∼1-10 mM), leading to the remaining challenge of selective detection and differentiation of endogenous H₂S in live biosystems. To this end, we quantitatively demonstrate that the thiolysis of NBD amine has much higher selectivity for H₂S over GSH than that of the reduction of aryl azide. Subsequently, we developed the first NBD-based cell-trappable probe 1 (AM-BODIPY-NBD) for highly selective and ultrasensitive imaging of intracellular H₂S. Probe 1 demonstrates a 207-fold fluorescence enhancement at 520 nm after reaction with H₂S/esterase to produce a bright BODIPY (quantum yield 0.42) and a detection limit of 15.7 nM. Probe 1 is water-soluble, cell-trappable, and not cytotoxic. Based on this excellent chemical tool, relative levels of basal H₂S in different cell lines were measured to reveal a positive correlation between endogenous H₂S and the metastatic potential of colon and breast cancer cells. In addition, H₂S biogenesis in vivo was also validated by probe 1 both in tobacco leaves under viral infection and in zebrafish after tail amputation. It is anticipated that probe 1 will have widespread applications in imaging and for investigating different H₂S-related biological processes and diseases.