Project description:Hydrogen sulfide (H2 S) is a gasotransmitter known to regulate physiological and pathological processes. Abnormal H2 S levels have been associated with a range of conditions, including Parkinson's and Alzheimer's diseases, cardiovascular and renal diseases, bacterial and viral infections, as well as cancer. Therefore, fast and sensitive H2 S detection is of significant clinical importance. Fluorescent H2 S probes hold great potential among the currently developed detection methods because of their high sensitivity, selectivity, and biocompatibility. However, many proposed probes do not provide a gold standard for proper use and selection. Consequently, issues arise when applying the probes in different conditions. Therefore, we systematically evaluated four commercially available probes (WSP-1, WSP-5, CAY, and P3), considering their detection range, sensitivity, selectivity, and performance in different environments. Furthermore, their capacity for endogenous H2 S imaging in live cells was demonstrated.
Project description:The new nonheme iron complexes FeII(BNPAPh2O)(N3) (1), FeIII(BNPAPh2O)(OH)(N3) (2), FeII(BNPAPh2O)(OH) (3), FeIII(BNPAPh2O)(OH)(NCS) (4), FeII(BNPAPh2O)(NCS) (5), FeIII(BNPAPh2O)(NCS)2 (6), and FeIII(BNPAPh2O)(N3)2 (7) (BNPAPh2O = 2-(bis((6-(neopentylamino)pyridin-2-yl) methyl)amino)-1,1-diphenylethanolate) were synthesized and characterized by single crystal X-ray diffraction (XRD), as well as by 1H NMR, 57Fe Mössbauer, and ATR-IR spectroscopies. Complex 2 was reacted with a series of carbon radicals, ArX3C· (ArX = p-X-C6H4), analogous to the proposed radical rebound step for nonheme iron hydroxylases and halogenases. The results show that for ArX3C· (X = Cl, H, tBu), only OH· transfer occurs to give ArX3COH. However, when X = OMe, a mixture of alcohol (ArX3COH) (30%) and azide (ArX3CN3) (40%) products was obtained. These data indicate that the rebound selectivity is influenced by the electron-rich nature of the carbon radicals for the azide complex. Reaction of 2 with Ph3C· in the presence of Sc3+ or H+ reverses the selectivity, giving only the azide product. In contrast to the mixed selectivity seen for 2, the reactivity of cis-FeIII(OH)(NCS) with the X = OMe radical derivative leads only to hydroxylation. Catalytic azidation was achieved with 1 as catalyst, λ3-azidoiodane as oxidant and azide source, and Ph3CH as test substrate, giving Ph3CN3 in 84% (TON = 8). These studies show that hydroxylation is favored over azidation for nonheme iron(III) complexes, but the nature of the carbon radical can alter this selectivity. If an OH· transfer pathway can be avoided, the FeIII(N3) complexes are capable of mediating both stoichiometric and catalytic azidation.
Project description:Two new coumarin-based "turn-off" fluorescent probes, (E)-3-((3,4-dihydroxybenzylidene)amino)-7-hydroxy-2H-chromen-2-one (BS1) and (E)-3-((2,4-dihydroxybenzylidene)amino)-7-hydroxy-2H-chromen-2-one (BS2), were synthesized and their detection of copper(II) and iron(III) ions was studied. Results show that both compounds are highly selective for Cu²? and Fe³? ions over other metal ions. However, BS2 is detected directly, while detection of BS1 involves a hydrolysis reaction to regenerate 3-amino-7-hydroxycoumarin (3) and 3,4-dihydroxybenzaldehyde, of which 3 is able to react with copper(II) or iron(III) ions. The interaction between the tested compounds and copper or iron ions is associated with a large fluorescence decrease, showing detection limits of ca. 10?? M. Preliminary studies employing epifluorescence microscopy demonstrate that Cu²? and Fe³? ions can be imaged in human neuroblastoma SH-SY5Y cells treated with the tested probes.
Project description:In the structure of the title compound, [Fe(C(7)H(7)N(2)O(2))(3)]·CH(3)CH(2)OH, the Fe(III) atom is in a distorted octa-hedral O(6) environment with the three hydroxamate O atoms (and the three carbonyl O atoms) arranged in a fac configuration and one of the hydroxamate ligands being puckered. The methyl C atom of the ethanol solvent mol-ecule is disordered over two positions with occupancies of 0.626?(13) and 0.374?(13), respectively. The cocrystallized ethanol mol-ecule is hydrogen bonded to one of the hydroxamate O atoms. O-H?O and N-H?O inter-actions generate infinite three-dimensional networks along [100], [010] and [001].
Project description:A series of fluorescent iron chelators has been synthesized such that a fluorescent function is covalently linked to a 3-hydroxypyridin-4-one. In the present study, the fluorescent iron chelators were loaded into isolated rat hepatocytes. The intracellular fluorescence was not only quenched by an addition of a highly lipophilic 8-hydroxyquinoline-iron(III) complex but also was dequenched by the addition of an excess of the membrane-permeable iron chelator CP94 (1,2-diethyl-3-hydroxypyridin-4-one). The time course of uptake of iron and iron chelation in single, intact cells was recorded on-line by using digital fluorescence microscopy. Intracellular concentrations of various fluorescent iron chelators were determined by using a spectrofluorophotometer subsequent to lysis of probe-loaded cells and were found to depend on their partition coefficients; the more hydrophobic the compound, the higher the intracellular concentration. An ex situ calibration method was used to determine the chelatable iron pool of cultured rat hepatocytes. CP655 (7-diethylamino-N-[(5-hydroxy-6-methyl-4-oxo-1,4-dihydropyridin-3-yl)methyl]-N-methyl-2-oxo-2H-chromen-3-carboxamide), which is a moderately lipophilic fluorescent chelator, was found to be the most sensitive probe for monitoring chelatable iron, as determined by the intracellular fluorescence increase induced by the addition of CP94. The concentration of the intracellular chelatable iron pool in hepatocytes was determined by this probe to be 5.4+/-1.3 microM.
Project description:Tricresyl phosphate (TCP) is an organophosphorous neurotoxin that has been detected in water, soil and air. Exposure to TCP in cockpit and cabin air poses a severe threat to flight safety and the health of the aircraft cabin occupants. Conventional methods for the detection of TCP in various samples are gas or liquid chromatography coupled to mass spectrometry, which are complex and expensive. To develop a simple low-cost methodology for the real-time monitoring of TCP in the environment, an effective catalyst is demanded for the hydrolysis of TCP under neutral condition. In this study, Ruthenium (III) hydroxide and Iron (III) hydroxide are found to facilitate the production of the alcoholysis and hydrolysis products of TCP, suggesting their role as a catalyst. With this finding, these metal hydroxides provide new potential to realize not only simple colorimetric or electrochemical detection of TCP, but also a simple detoxication strategy for TCP in environment. In addition, the catalytic capability of Ru (III) or Fe (III) hydroxide for TCP gives a hint that they can potentially serve as catalysts for the hydrolysis of alcolyolysis of many other organophosphate compounds.
Project description:The title compound, [Fe(C(12)H(8)N(2))(3)][Fe(2)Cl(6)O]·0.5CH(3)CH(2)OH, consists of one [Fe(phen)(3)](2+) cation (phen = 1,10-phen-anthroline), one [Fe(2)Cl(6)O](2-) anion and one half-mol-ecule of ethanol. In the cation, the Fe(II) atom is coordinated by six N atoms from three phen ligands in a distorted octa-hedral geometry. In the bent anion, two Fe(III) atoms are connected by a bridging oxide O atom [bridging angle = 160.6 (4)°], and each Fe(III) atom is also coordinated by three Cl atoms, completing a distorted tetra-hedral geometry.
Project description:The development of synthetic lanthanide luminescent probes for selective sensing or binding anions in aqueous medium requires an understanding of how these anions interact with synthetic lanthanide probes. Synthetic lanthanide probes designed to differentiate anions in aqueous medium could underpin exciting new sensing tools for biomedical research and drug discovery. In this direction, we present three mononuclear lanthanide-based complexes, EuLCl3 (1), SmLCl3 (2), and TbLCl3 (3), incorporating a hexadentate aminomethylpiperidine-based nitrogen-rich heterocyclic ligand L for sensing anion and establishing mechanistic insight on their binding activities in aqueous medium. All these complexes are meticulously studied for their preferential selectivities towards different anions such as HPO42-, SO42-, CH3COO-, I-, Br-, Cl-, F-, NO3-, CO32-/HCO3-, and HSO4- at pH 7.4 in aqueous HEPES (2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid) buffer. Among the anions scanned, HPO42- showed an excellent luminescence change with all three complexes. Job's plot and ESI-MS support the 1:2 association between the receptors and HPO42-. Systematic spectrophotometric titrations of 1-3 against HPO42- demonstrates that the emission intensities of 1 and 2 were enhanced slightly upon the addition of HPO42- in the range 0.01-1 equiv and 0.01-2 equiv., respectively. Among the three complexes, complex 3 showed a steady quenching of luminescence throughout the titration of hydrogen phosphate. The lower and higher detection limits of HPO42- by complexes 1 and 2 were determined as 0.1-4 mM and 0.4-3.2 mM, respectively, while complex 3 covered 0.2-100 μM. This concludes that all complexes demonstrated a high degree of sensitivity and selectivity towards HPO42-.
Project description:Fluorescent dyes are widely used in the detection of labile (free or exchangeable) Zn(2+) and Ca(2+) in living cells. However, their specificity over other cations and selectivity for detection of labile vs. protein-bound metal in cells remains unclear. We characterized these important properties for commonly used Zn(2+) and Ca(2+) dyes in a cellular environment. By tracing the fluorescence emission signal along with UV-Vis and size exclusion chromatography-inductively coupled plasma mass spectrometry (SEC-ICP-MS) in tandem, we demonstrated that among the dyes used for Zn(2+), Zinpyr-1 fluoresces in the low molecular mass (LMM) region containing labile Zn(2+), but also fluoresces in different molecular mass regions where zinc ion is detected. However, FluoZin™-3 AM, Newport Green™ DCF and Zinquin ethyl ester display weak fluorescence, lack of metal specificity and respond strongly in the high molecular mass (HMM) region. Four Ca(2+) dyes were studied in an unperturbed cellular environment, and two of these were tested for binding behavior under an intracellular Ca(2+) release stimulus. A majority of Ca(2+) was in the labile form as tested by SEC-ICP-MS, but the fluorescence from Calcium Green-1™ AM, Oregon Green® 488 BAPTA-1, Fura red™ AM and Fluo-4 NW dyes in cells did not correspond to free Ca(2+) detection. Instead, the dyes showed non-specific fluorescence in the mid- and high-molecular mass regions containing Zn, Fe and Cu. Proteomic analysis of one of the commonly seen fluorescing regions showed the possibility for some dyes to recognize Zn and Cu bound to metallothionein 2. These studies indicate that Zn(2+) and Ca(2+) binding dyes manifest fluorescence responses that are not unique to recognition of labile metals and bind other metals, leading to suboptimal specificity and selectivity.
Project description:Redox processes are involved in almost every cell of the body as a consequence of aerobic life. In the past decades, redox biology has been increasingly recognized as one of the key themes in cell signaling. The progress has been accelerated by development of fluorescent probes that can monitor redox conditions and dynamics in cells and cell compartments. This short paper focuses on fluorescent redox probes that are genetically encoded, and discusses their properties, molecular mechanism, advantages and pitfalls. Our recent work on reaction-based encoded probes that are responsive to particular redox signaling molecules is also reviewed. Future challenges and directions are also commented.