Cysteine triggered cascade reaction forming coumarin: Visualization of cysteine fluctuation in alcoholic liver disease by a NIR fluorescent probe.

Alcoholic liver disease (ALD) is a chronic toxic liver injury caused by long-term heavy drinking. Due to the increasing incidence, ALD is becoming one of important medical tasks. Many studies have shown that the main mechanism of liver damage caused by large amounts of alcohol may be related to antioxidant stress. As an important antioxidant, cysteine (Cys) is involved in maintaining the normal redox balance and detoxifying metabolic function of the liver, which may be closely related to the pathogenesis of ALD. Therefore, it is necessary to develop a simple non-invasive method for rapid monitoring of Cys in liver. Thus, a near-infrared (NIR) fluorescent probe DCI-Ac-Cys which undergoes Cys triggered cascade reaction to form coumarin fluorophore is developed. Using the DCI-Ac-Cys, decreased Cys was observed in the liver of ALD mice. Importantly, different levels of Cys were monitored in the livers of ALD mice taking silybin and curcumin with the antioxidant effects, indicating the excellent therapeutic effect on ALD. This study provides the important references for the accurate diagnosis of ALD and the pharmacodynamic evaluation of silybin and curcumin in the treatment of ALD, and support new ideas for the pathogenesis of ALD.

The first ER-targeting flavone-based fluorescent probe for Cys: Applications in real-time tracking in an epilepsy model and food analysis.

Epilepsy is one of the most commonly-seen neurological disorders, and both endoplasmic reticulum stress (ERS) and oxidative stress (OS) have been demonstrated to be associated with epileptic seizures. As one of the three endogenous thiol-containing amino acids, cysteine (Cys) is recognized not only as an important biomarker of various biological processes but also widely used as a significant additive in the food industry. However, the exact role that Cys plays in ERS has not been well answered up to now. In this paper, we reported the first flavone-based fluorescent probe (namely BFC) with nice endoplasmic reticulum (ER)-targeting ability, which was capable of monitoring Cys in a fast response (3.0 min), large stokes shift (130 nm) and low detection limit (10.4 nM). The recognition mechanism of Cys could be attributed to the addition-cyclization reaction involving a Cys residue and an acrylate group, resulting in the release of the strong excited-state intramolecular proton transfer (ESIPT) emission molecule of benzoflavonol (BF). The low cytotoxicity and good biocompatibility of the probe BFC allowed for monitoring the fluctuation of endogenous Cys levels under both ERS and OS processes, as well as in zebrafish models of epilepsy. Quantitative determination of Cys with the probe BFC was also achieved in three different food samples. Additionally, a probe-immersed test strips integrated with a smartphone device was successfully constructed for on-site colorimetric detection of Cys. Undoubtedly, our work provided a valuable tool for tracking Cys levels in both an epilepsy model and real food samples.

A novel structurally modified isophorone fluorescent probe for H2S detection.

Hydrogen sulfide (H2S) has a comprehensive contribution to the normal operation and stability of organisms and is also present in environmental water samples and food deterioration. Thus, it is exceedingly promising and significant to develop a highly sensitive detection technique for tracing H2S. Inspired by this, we designed and synthesized a new fluorescent probe 2-[3-[2-[3-bromo-4-(2,4- dinitrobenzenesulfonate)] ethenyl]-5,5-dimethyl-2-cyclohexen-1-ylidene]propanedinitrile (SP-Br) for hydrosulfide ion detection by introducing bromine atom. Compared with reported H2S probes based on the same fluorescent parent, SP-Br has longer fluorescence emission (λem = 670 nm), shorter response time (3 min), lower detection limit (149 nM), and wider detection range (0-30 nM). SP-Br can emit weak yellow fluorescence, and the emission intensity at 670 nm is considerably enhanced in the presence of hydrosulfide ions. The identification mechanism of hydrosulfide ion by SP-Br was verified by high-resolution mass spectrometry, fluorescence, and UV-vis absorption spectroscopy. In addition, SP-Br has been successfully applied to the monitoring of actual water samples and beer samples and has certain development prospects and value in the fields of environmental pollution and food quality analysis.

Fluorescence based metabisulfite sensing: New aspects of ion sensing by a styryl benzothiazolium dye and understanding nitrite interference.

Detection of specific ions using fluorescent probes has relevance in several areas of therapeutics development and environmental science. Here, we provide new perspectives to the sensing of a styryl benzothiazolium-based fluorescent compound 1 and report that sensing properties are for sulfite ions in general with highest preference for metabisulfite ions (S2O52-) adding to its previously determined role as a bisulfite ion sensor. This probe exhibits its sensing action via an addition reaction in which the styryl double bond gets reduced. The interference studies highlighted that the sequence of addition of nitrite and metabisulfite has a bearing on the overall interference outcome. Spectroscopic studies revealed that the order of preferential sensing of sulfites and sulfide ion is S2O52- > HSO3- > SO32- > S2-. Although this probe displays robust sensing on its own through fluorescence quenching, its fluorescence emission can be enhanced at much lower concentrations in the presence of a G-quadruplex DNA without compromising the outcome of the sensing.

A multifunctional near-infrared fluorescent probe based on benzothiazole structure for fluoride-ion detection.

Fluoride ions (F-) are one of the essential trace elements for the human body and are widely existed in nature. In this study, we present a novel fluorescent probe (YF-SZ-F) designed and synthesized for the specific detection of F-. The probe exhibits high sensitivity, excellent selectivity, and low cytotoxicity, making it a promising tool for biomedical applications. Imaging experiments conducted on zebrafish and Arabidopsis roots demonstrate the probe's remarkable cellular permeability and biocompatibility, laying a solid foundation for its potential biomedical utility. Furthermore, the probe holds potential for practical applications in environmental monitoring and public health through its capability to detect fluoride ions in water samples and via mobile phone software. This multifaceted functionality underscores the broad applicability and significance of the fluorescent probe, not only in scientific research but also in real-world scenarios, contributing to the development of more convenient and precise methods for fluoride detection.

A near-infrared ratiometric fluorescent probe for the sensing and imaging of sulfur dioxide derivatives in living systems.

As a reactive sulfur species, sulfur dioxide (SO2) and its derivatives play crucial role in various physiological processes, which can maintain redox homeostasis at normal levels and lead to the occurrence of many diseases at abnormal levels. So, the development of a suitable fluorescent probe is a crucial step in advancing our understanding of the role of SO2 derivatives in living organisms. Herein, we developed a near-infrared fluorescent probe (SP) based on the ICT mechanism to monitor SO2 derivatives in living organisms in a ratiometric manner. The probe SP exhibited excellent selectivity, good sensitivity, fast response rate (within 50 s), and low detection limit (1.79 µM). In addition, the cell experiment results suggested that the SP has been successfully employed for the real-time monitoring of endogenous and exogenous SO2 derivatives with negligible cytotoxicity. Moreover, SP was effective in detecting SO2 derivatives in mice.

Fluorescent probes based on N-CQDs: For direct detection of food additives STPP and Al3.

Sodium tripolyphosphate (STPP), as one of the many food additives, can cause gastrointestinal discomfort and a variety of adverse reactions when ingested by the human body, which is a great potential threat to human health. Therefore, it is necessary to develop a fast, sensitive and simple method to detect STPP in food. In this study, we synthesized a kind of nitrogen-doped carbon quantum dots (N-CQDs), and were surprised to find that the addition of STPP led to the gradual enhancement of the emission peaks of the N-CQDs, with a good linearity in the range of 0.067-1.96 µM and a low detection limit as low as 0.024 µM. Up to now, there is no report on the use of carbon quantum dots for the direct detection of STPP. Meanwhile, we found that the addition of Al3+ effectively bursts the fluorescence intensity of N-CQDs@STPP solution and has a good linear relationship in the range of 0.33-6.25 µM with a lower detection limit of 0.24 µM. To this end, we developed a fluorescent probe to detect STPP and Al3+. In addition, the probe was successfully applied to the detection of bread samples, which has great potential for practical application.

Selective, Intrinsically Fluorescent Trk Modulating Probes.

Neurotrophins (NTs) elicit the growth, survival, and differentiation of neurons and other neuroectoderm tissues via activation of Trk receptors. Hot spots for NT·Trk interactions involve three neurotrophin loops. Mimicry of these using "cyclo-organopeptides" comprising loop sequences cyclized onto endocyclic organic fragments accounts for a few of the low molecular mass Trk agonists or modulators reported so far; the majority are nonpeptidic small molecules accessed without molecular design and identified in random screens. It has proven difficult to verify activities induced by low molecular mass substances are due to Trk activation (rather than via other receptors), enhanced Trk expression, enhanced NT expression, or other pathways. Consequently, identification of selective probes for the various Trk receptors (e.g., A, B, and C) has been very challenging. Further, a key feature of probes for early stage assays is that they should be easily detectable, and none of the compounds reported to date are. In this work, we designed novel cyclo-organopeptide derivatives where the organic fragment is a BODIPY fluor and found ones that selectively, though not specifically, activate TrkA, B, or C. One of the assays used to reach this conclusion (binding to live Trk-expressing cells) relied on intrinsic fluorescence in the tested materials. Consequently, this work established low molecular mass Trk-selective probes exhibiting neuroprotective effects.

Novel benzothiazole-based fluorescent probe for efficient detection of Cu2+/S2- and Zn2+ and its applicability in cell imaging.

BACKGROUND: In recent years, environmental pollution has been increasing due to the excessive emission of toxic ions, which has caused serious harm to human health and ecological environment. There are various methods for detecting Cu2+, S2- and Zn2+, but the traditional ion detection methods have obvious disadvantages, such as poor selectivity and long detection time. Therefore, it is still crucial to develop simple, efficient and rapid detection methods. RESULTS: A fluorescent probe based on benzothiazole, (E)-N'-(3-(benzo[d]thiazol-2-yl)-2-hydroxy-5-methylbenzylidene)-3,4,5-tris(benzyloxy)benzohydrazide (BT), was designed and synthesized. It was characterized using ESI-MS, 1H NMR, and 13C NMR. BT can be used as a chemosensor to detect Cu2+, S2- and Zn2+ in CH3CN/H2O (7:3, v/v, pH = 7.4, HEPES buffer: 0.1 M), with detection limits of 0.301 µM, 0.017 µM, and 0.535 µM, respectively. At an excitation wavelength of 320 nm, BT exhibits an "on-off-on" response to Cu2+/S2- and enhanced fluorescence response to Zn2+, with a change in fluorescence color from orange to green. The coordination ratio of ions to the probe was determined to be 1:1 through Job's plot and hydrogen spectral titration. The recognition mechanism was discussed in conjunction with theoretical calculations. Furthermore, the probe has been successfully used in test strips and medical swabs colorimetry, as well as live cell imaging. SIGNIFICANCE: The probe BT lays the foundation for the design and synthesis of multifunctional fluorescent probes. As a portable detection method, probe BT was used to detect Cu2+, S2- and Zn2+ on strips. Furthermore, the probe was applied to biological cells to detect target ions with low cytotoxicity and excellent cell permeability. This indicating that it can be used as a potential candidate for tracking Cu2+ and S2- in clinical diagnostics and biological systems.

Functionalized α-Cyanostilbene Derivatives for Detection of Hypoxia or Proteostasis Imbalance in Live Cells.

α-Cyanostilbene represents one of the easily functionalized aggregation-induced emission (AIE) scaffolds. It has been widely adopted for the construction of fluorescent materials for broad applications. Here, we further expanded the utilization of α-cyanostilbene derivatives for the detection of hypoxia or proteostasis imbalance in live cells. Four different amine containing donors were introduced to construct α-cyanostilbene derivatives (R-ASC) with donor-acceptor scaffolds. Equipped with the cysteine (Cys) reactive group, maleimide (MI), R-ASC-MI shows fluorescence turn-on property upon binding with unfolded proteins in vitro and in live cells under proteostatic stress. By virtue of R-ASC-MI, the level of unfolded protein loads in cells can be quantified by flow cytometry, or visualized under microscope. Furthermore, we also characterized the performance of R-ASC-NO2, synthetic precursors of R-ASC-MI, in cellular hypoxia. R-ASC-NO2 revealed upregulated activities of nitroreductase, as well as increased hydrophobicity in live cells, under either chemical (NaN3) induced or atmospheric (1% O2) hypoxia. Together, the advantages of easy modification and high signal-to-noise ratio of new α-cyanostilbene derivatives reported in this work highlight the great potential of α-cyanostilbene in constructing functional biosensors and many other domains.

An activity-based sensing fluorogenic probe for monitoring O-methyltransferase in plants.

Activity-based sensing probes are powerful tools for monitoring enzymatic activities in complex biological samples such as cellular and live animals; however, their application in plants remains challenging. Herein, fourteen activity-based fluorescent probes were assayed against Arabidopsis O-methyltransferases (AtOMTs). One probe, 3-BTD, displayed a high selectivity, reactivity, and fluorescence response toward AtOMTs especially the isoform AtCCoAOMT. We further characterized the features of this probe and explored whether it could be used to detect OMT activities in living plant cells. Our results show that 3-BTD can be used to visualize OMT activity in Arabidopsis, and no fluorescent signal was observed in the comt/ccoaomt double mutant, indicating that it has good specificity. Interestingly, in contrast to the observation that AtCCoAOMT-YFP accumulated in both cytoplasm and nucleus, OMT enzymatic activity tracked by 3-BTD probe was found only in the cytoplasm. This underscores the importance of activity-based sensing in studying protein function. Moreover, 3-BTD can be successfully applied in OMT visualization of different plants. This study indicates that 3-BTD can serve as a potential probe for in situ monitoring the real activity of OMT in multiple plants and provides a strategy for visualizing the activity of other enzymes in plants.

Simultaneous detection of cysteine and glutathione in food with a two-channel near-infrared fluorescent probe.

L-Cysteine (Cys) and glutathione (GSH) are closely related biological species that widely exist in food and living cells. To simultaneously detect Cys and GSH from different emission channels, we developed a fluorescent probe (BDP-NBD) based on near-infrared BODIPY and 7-nitrobenzofurazan (NBD). Upon nucleophilic substitution reaction with GSH, BDP-NBD generated an emission band at 713 nm, which can be used to determine GSH (0-100 µM) with a low detection limit (34 nM). Different from GSH, BDP-NBD underwent a nucleophilic substitution-rearrangement reaction with Cys, affording two emission bands at 550 nm and 713 nm, respectively. BDP-NBD was successfully employed to quantify Cys and GSH in various food samples with good recoveries (86.6%-104.6%). Besides, BDP-NBD can image Cys and GSH in living cells from two emission channels. Therefore, this work developed a tool for the simultaneous determination of Cys and GSH in both food and living cells so as to ensure food safety and human health.

An activatable near-infrared fluorescent probe with large Stokes shift for visualizing peroxynitrite in Alzheimer's disease models.

Alzheimer's disease (AD), characterized by its incurable nature and prevalence among the elderly, has remained a focal point in medical research. Increasing evidence suggests that peroxynitrite (ONOO-) serves as a crucial biomarker for the diagnosis of AD. In this study, we present a novel, easily available, high-yield, and cost-effective near-infrared (NIR) fluorescent probe, CDCI-ONOO. This probe utilizes a coumarin-dicyanoisophorone conjugate as the fluorophore and diphenylphosphinic chloride as the recognition site, enabling the detection of ONOO- both in vitro and in vivo. Upon interaction with ONOO-, CDCI-ONOO exhibits a distinct maximum emission peak at 715 nm with a substantial Stokes shift of 184 nm. The probe demonstrates excellent selectivity and sensitivity (LOD = 144 nM), along with noticeable colorimetric and fluorescence changes after the reaction. Comprehensive analyses using high-performance liquid chromatography (HPLC), high-resolution mass spectrometry (HRMS), and density functional theory (DFT) calculations confirm that the reaction with ONOO- restores the initially quenched Intramolecular Charge Transfer (ICT), resulting in the formation of CDCI-OH, a product that emitting fluorescence in the near-infrared region. Furthermore, we demonstrated the successful application of CDCI-ONOO for ONOO- detection in neuronal cells and imaging of ONOO- in the brains of mice. These findings underscore the potential of CDCI-ONOO as a near-infrared fluorescent probe for in vivo ONOO- detection, offering a significant avenue for advancing our understanding of AD pathology and diagnosis.

A fluorescent probe for monitoring carboxylesterases in pulmonary cells under permissive hypercapnia condition.

Herein, by combining the benzofuranone-derived fluorophore and the carbamate recognition group, a fluorescent probe named BFO-CarE was developed for monitoring the carboxylesterase (CarE) level in pulmonary cells under the permissive hypercapnia condition. It showed a notable fluorescence response towards CarE at 570 nm under the excitation of 510 nm. The in-solution tests revealed the advantages of BFO-CarE including high sensitivity, high specificity, relatively rapid response, and high steadiness. It was also low-toxic upon the pulmonary cell lines. During the intracellular imaging in pulmonary cells, BFO-CarE achieved the monitoring of the CarE level in both inhibition and activation status. In particular, BFO-CarE realized the visualization of the affection of the permissive hypercapnia condition on the CarE level, which indicated the hypoxia tolerance of CarE. This work was informative for investigating the impact of hypoxia in pulmonary cells, and the corresponding anaesthesia-related approaches.

Diagnostic value for methylation in cervical cancer based on a small-molecule fluorescent probe targeting DNMT1.

BACKGROUND: Value analysis of a small-molecule fluorescent probe for methylation detection in different cervical lesions. MATERIALS AND METHODS: (1) The grayscale values of distinct lesion tissues were remarkably distinct among the four groups (p < 0.05). The comparison of the grayscale value between the two groups showed that the CA group noticeably exceeded the LSIL and cervicitis groups, and the HSIL group was apparently higher than the LSIL and cervicitis groups (p < 0.05); (2) The mean grayscale values of the enrolled subjects were calculated with 55.21 as the midline, with >55.21 as positive and ≤55.21 as negative. RESULTS: The results showed that the positive rate of the cervicitis group was 0.00%, the LSIL group 67.74%, the HSIL group 83.33%, and the CA group 100.00%. The results among the four groups were notably distinct (p < 0.05); (3) The comparison among DAPI, probe, bright, and merged images of cervicitis, LSIL, HSIL, and CA indicated that different cervical lesions were with quite various stains. CONCLUSION: The grayscale value, positive rate, and stained picture of distinct cervical lesions were remarkably different. The small-molecule fluorescent probe has a good value in differentiating cervical lesions and can be considered for popularization and application.

A Review on Recent Development of Phenothiazine-Based Chromogenic and Fluorogenic Sensors for the Detection of Cations, Anions, and Neutral Analytes.

This review provides an in-depth examination of recent progress in the development of chemosensors, with a particular emphasis on colorimetric and fluorescent probes. It systematically explores various sensing mechanisms, including metal-to-ligand charge transfer (MLCT), ligand-to-metal charge transfer (LMCT), photoinduced electron transfer (PET), intramolecular charge transfer (ICT), and fluorescence resonance energy transfer (FRET), and elucidates the mechanism of action for cation and anion chemosensors. Special attention is given to phenothiazine-based fluorescence probes, highlighting their exceptional sensitivity and rapid detection abilities for a broad spectrum of analytes, including cations, anions, and small molecules. Phenothiazine chemosensors have emerged as versatile tools widely employed in a multitude of applications, spanning environmental and biomedical fields. Furthermore, it addresses existing challenges and offers insights into future research directions, aiming to facilitate the continued advancement of phenothiazine-based fluorescent probes.

Rational design of a high-affinity fluorescent probe for visualizing monitoring the amyloid ß clearance effect of anti-Alzheimer's disease drug candidates.

Beta-amyloid (Aß), the most pivotal pathological hallmark for Alzheimer's disease (AD) diagnosis and drug evaluation, was recognized by TZ095, a high-affinity fluorescent probe developed by rational molecular design. With a TICT mechanism, TZ095 exhibited remarkable affinity with Aß aggregates (Kd = 81.54 nM for oligomers; Kd = 66.70 nM for fibril) and substantial fluorescence enhancement (F/F0 = 44), enabling real-time monitoring of Aß in live cells and nematodes. Significantly, this work used TZ095 to construct a new protocol that can quickly and conveniently monitor Aß changes at the cellular and nematode levels to evaluate the anti-AD efficacy of candidate compounds, and four reported Aß-lowering drug candidates were administrated for validation. Imaging data demonstrated that TZ095 can visually and quantitatively track the effect of Aß elimination after drug treatment. Furthermore, TZ095 excelled in ex vivo histological staining of 12-month-old APP/PS1 mouse brains, accurately visualizing Aß plaques. Integrating CUBIC technology, TZ095 facilitated whole-brain, 3D imaging of Aß distribution in APP/PS1 mice, enabling high-resolution in situ analysis of Aß plaques. Collectively, these innovative applications of TZ095 offer a promising strategy for rapid, convenient, and real-time monitoring of Aß levels in preclinical therapeutic assessments.

A novel ultrafast and highly sensitive NIR fluorescent probe for the detection of organophosphorus pesticides in foods and biological systems.

The threat posed by organophosphorus pesticides (OPS) to food safety, human health, and the ecological environment is significant, which underscoring the need for the development of new detection tools. We designed and synthesized a NIR fluorescent probe PT-CES which targets carboxylesterase (CES), for the detection of OPS based on the principle of enzyme inhibition. The PT-CES is capable of instantaneous response to CES, exhibiting excellent stability, anti-interference capability. PT-CES realizes the quantitative detection of CES and OPS. It is noteworthy that PT-CES shows excellent stable and accurate detection ability in vegetable pesticide testing. It also enables the monitoring of CES activity in cells and liver tissue. This provides a novel tool for tracking the effect of OPS on CES activity in biological systems. Furthermore, it provides a useful method for ensuring food safety and enhancing pesticide residue analysis.

Developing a vanillin-derived imidazo-pyridin-containing fluorescent probe for imaging cysteine in living pulmonary cells under oxygen supply variation.

In this work, derived from vanillin and imidazo-pyridin backbone, a fluorescent probe IPV-Cys was developed for imaging the cysteine (Cys) level in living pulmonary cells under oxygen supply variation. By mimicking the oxygen supply variation in both the solution test and cellular imaging, the optical performance and imaging effect of IPV-Cys was investigated. In the solution system, the oxygen supply variation caused no impact on the reporting signals. The fluorescence reporting signal intensity at 490 nm suggested the enhancement along with the increase of the Cys concentration. The advantages of IPV-Cys included relatively high sensitivity, high stability, and high selectivity. On the basis of the low cyto-toxicity, IPV-Cys achieved the monitoring the endogenous Cys level in in living pulmonary cells and the impact of the oxygen supply variation by reporting fluorescence signals. The information here was meaningful for both the pre-clinical diagnosis and surgical techniques.

High-fidelity imaging of drug-induced acute gastritis by using a fluorescent and photoacoustic dual-modal probe with good stability in stomach acid.

In consideration of deep tissue imaging and signal fidelity, fluorescent-photoacoustic (PA) dual-modal probes are much more desirable. However, dual-modal imaging of gastritis using molecular probes remains a challenge due to the harsh gastric acid environment in the stomach. Based on the positive correlation between gastritis and cell viscosity, stomach acid-stable and viscosity-activated probes could potentially diagnose gastritis. As a proof of concept, herein, a fluorescent and photoacoustic dual-modal probe (named WSP-1) is revealed for the imaging of drug-induced acute gastritis in vivo. WSP-1 exhibits viscosity-dependent fluorescence emission and photoacoustic signals. A rotatable C-C single bond is incorporated into the D-π-A structure of WSP-1, which could facilitate the formation of the twisted intramolecular charge transfer (TICT) state in a low-viscosity environment (weak fluorescence/PA signal) and the intramolecular charge transfer (ICT) state in a high-viscosity environment (strong fluorescence/PA signal). WSP-1 has demonstrated the capability to target mitochondria and can be utilized to monitor the viscosity enhancement of cells during inflammation. Most importantly, WSP-1 exhibits good optical and structural stability in gastric acid. By leveraging these desirable features of WSP-1, we have achieved fluorescent and 3D photoacoustic in situ imaging of drug-induced acute gastritis following oral administration of WSP-1.