Diagnosis of Alzheimer's Disease and In Situ Biological Imaging via an Activatable Near-Infrared Fluorescence Probe.

Alzheimer's disease (AD) is a common neurodegenerative disease that makes the brain nervous system degenerate rapidly and is accompanied by some special cognitive and behavioral dysfunction. Recently, butyrylcholinesterase (BChE) was reported as an important enzyme, whose activity can provide predictive value for timely discovery and diagnosis of AD. Therefore, it is indispensable to design a detection tool for selective and rapid response toward BChE. In this study, we developed a novel near-infrared fluorescent probe (Chy-1) for the detection of BChE activity. An excellent sensitivity, good biocompatibility, and lower limit of detection (LOD) of 0.12 ng/mL made the probe extremely specific for BChE, which was successfully used in biological imaging. What is more, Chy-1 can not only clearly distinguish tumor from normal cells but also forms a clear boundary between the normal and cancer tissues due to the obvious difference in fluorescence intensity produced via in situ spraying. Most important of all, Chy-1 was also successfully applied to track the BChE activity in AD mouse models. Based on this research, the novel probe may be a powerful tool for clinical diagnosis and therapy of tumor and neurodegenerative diseases.

A Novel Fluorescent Probe Strategy Activated by ß-Glucuronidase for Assisting Surgical Resection of Liver Cancer.

Liver cancer is a primary malignant tumor with a very high fatality rate, which has seriously threatened human health and life. In normal hepatocellular lesions, ß-glucuronidase (GLU) activity in liver cancer tissues is significantly increased. Therefore, GLU has become one of the important biomarkers of primary liver cancer. Here, a series of fluorescent probes (DCDH, DCDCH3, DCDOCH3, and DCDNO2) for early diagnosis of liver cancer and auxiliary surgical resection were successfully synthesized. Since the electron-withdrawing group -NO2 connected to the probe DCDNO2 accelerates the rapid cleavage of the glycosidic bond, DCDNO2 exhibits superior fluorescence properties that are more sensitive and rapid than the other three probes DCDH, DCDCH3, and DCDOCH3 when detecting GLU. DCDNO2 has been well-applied in real-time fluorescent visualization imaging for the detection of GLU activity in liver cancer cells and tumor tissues. In addition, DCDNO2 has also been successfully used in the early diagnosis of liver cancer and real-time imaging to guide the surgical resection of liver cancer tumors. Therefore, DCDNO2 has great potential for development in bioclinical medicine for the early detection and treatment of liver cancer.

Nanoscale Metal-Organic Layers Detect Mitochondrial Dysregulation and Chemoresistance via Ratiometric Sensing of Glutathione and pH.

Mitochondrial dysregulation controls cell death and survival by changing endogenous molecule concentrations and ion flows across the membrane. Here, we report the design of a triply emissive nanoscale metal-organic layer (nMOL), NA@Zr-BTB/F/R, for sensing mitochondrial dysregulation. Zr-BTB nMOL containing Zr6 secondary building units (SBUs) and 2,4,6-tris(4-carboxyphenyl)aniline (BTB-NH2) ligands was postsynthetically functionalized to afford NA@Zr-BTB/F/R by exchanging formate capping groups on the SBUs with glutathione(GSH)-selective (2E)-1-(2'-naphthyl)-3-(4-carboxyphenyl)-2-propen-1-one (NA) and covalent conjugation of pH-sensitive fluorescein (F) and GSH/pH-independent rhodamine-B (R) to the BTB-NH2 ligands. Cell imaging demonstrated NA@Zr-BTB/F/R as a ratiometric sensor for mitochondrial dysregulation and chemotherapy resistance via GSH and pH sensing.

Long-Wavelength Ratiometric Fluorescent Probe for the Early Diagnosis of Diabetes.

Diabetes is a metabolic disease caused by high blood sugar. Patients are often suffering from high blood pressure and arteriosclerosis, which may even evolve into liver disease, kidney disease, and other diabetic complications. Dipeptide peptidase IV (DPP-IV) plays an important role in regulating blood sugar levels and is one of the targets for the diagnosis and treatment of diabetes. Here, a long-wavelength ratiometric fluorescent probe DCDHFNH2-dpp4 for detecting DPP-IV was designed and synthesized. DCDHFNH2-dpp4 was used to detect DPP-IV in healthy, tumor-bearing, and diabetic mice, and only diabetic mice showed strong fluorescence signals. In organ imaging, it is found that DPP-IV is relatively enriched in the liver of diabetic mice. In addition, probe DCDHFNH2-dpp4 also exhibited a significant ratiometric fluorescence signal in the serum of diabetic mice. Therefore, the fluorescent probe DCDHFNH2-dpp4 has shown outstanding potential in the early diagnosis of diabetes, and DCDHFNH2-dpp4 is hopeful to be applied to actual clinical medicine.

A naphthalimide-based lysosome-targeting fluorescent probe for the selective detection and imaging of endogenous peroxynitrite in living cells.

The morpholine (ML) group can be used as a targeting unit for lysosomes. Here, a novel turn-off fluorescence probe for the highly selective imaging of peroxynitrite (ONOO-) produced by the endogenous stimulation of lysosomes in living cells is presented. The probe, denoted ML-NAP-DPPEA, comprises ML and 2-(diphenylphosphino)ethylamine (DPPE) groups attached to the fluorophore naphthalimide (NAP). ML-NAP-DPPEA shows excellent properties, including high selectivity for ONOO-, low cytotoxicity, and no interference, leading to low detection limits (17.6 nM). In the presence of ONOO-, the secondary amine group (NH) is oxidized to an electron-withdrawing group (HN → O), which quenches the fluorescence of ML-NAP-DPPEA. This intracellular lysosomal imaging technique was tested, and the results pointed to its potential use as a probe for studying the biological function and pathological effects of ONOO- in subcellular structure. Graphical abstract.

Dually emitting carbon dots as fluorescent probes for ratiometric fluorescent sensing of pH values, mercury(II), chloride and Cr(VI) via different mechanisms.

The authors describe the preparation of carbon dots (CDs) that display both blue (~ 410 nm) and yellow (~ 565 nm) emission peaks. The CDs was synthesized by solvothermal treatment of o-phenylenediamine in aqueous ethyl alcohol at pH ~7.0. The CDs are shown to be useful fluorescent probes for pH values in that the ratio of fluorescences at 565 and 410 nm strongly depends on the pH value in the range from 4.5-6.5 and 10.0-13.0, respectively. The blue fluorescence is quenched by 91% by 100 µM solutions of Hg(II) through an electron transfer process, and is restored by 97% an addition of chloride (0.5 mM). The yellow fluorescence, in contrast, is hardly affected. The ratio of fluorescences at 414 and 565 nm drops linearly in the 30 to 60 µM of Hg(II) concentration range, and the limit of detection is 60 nM. Fluorescence is linearly restored in the 70 to 180 µM chloride concentration range, and the LOD is 2.8 nM. Both the blue and the yellow emission are reduced by Cr(VI) (chromate) due to an inner filter effect at pH 3.0. The ratio of fluorescences (410/565 nm) drops linearly in the 20 to 250 µM Cr(VI) concentration range, and the LOD is 260 nM. The method was utilized to analysis of chloride in salt lake brine and of Cr(VI) in spiked tap water. Graphical abstract Schematic presentation of carbon dots with pH-dependent dual emission (at ~ 410 nm and ~ 565 nm). They are shown to be viable fluorescent probes for ratiometric sensing of pH values, mercury(II), chloride and Cr(VI) via different mechanisms.

A Phenylselenium-Substituted BODIPY Fluorescent Turn-off Probe for Fluorescence Imaging of Hydrogen Sulfide in Living Cells.

Herein a phenylselenium-substituted BODIPY (1) fluorescent turn-off sensor was developed for the purpose to achieve excellent selectivity and sensitivity for H2S detection based on the substitution reaction of the phenylselenide group at the 3-position with H2S. The excess addition of hydrogen sulfide promoted further substitution of the phenylselenide group at the 5-position of the probe and was accompanied by a further decrease in fluorescence emission intensity. Sensor 1 demonstrated remarkable performance with 49-fold red color fluorescence intensity decrease at longer excitation wavelength, a low detection limit (0.0025 µM), and specific fluorescent response toward H2S over anions, biothiols, and other amino acids in neutral media. It showed no obvious cell toxicity and good membrane permeability, which was well exploited for intracellular H2S detection and imaging through fluorescence microscopy imaging.

Fast and Selective Two-Stage Ratiometric Fluorescent Probes for Imaging of Glutathione in Living Cells.

Two fluorescent, m-nitrophenol-substituted difluoroboron dipyrromethene dyes have been designed by nucleophilic substitution reaction of 3,5-dichloro-4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY). Nonsymmetric and symmetric probes, that is. BODIPY 1 (with one nitrophenol group at the position 3) and BODIPY 2 (with two nitrophenol groups at the positions 3 and 5) were applied to ratiometric fluorescent glutathione detection. The detection is based on the two-step nucleophilic aromatic substitution of the nitrophenol groups of the probes by glutathione in buffer solution containing CTAB. In the first stage, probe 1 showed ratiometric fluorescent color change from green (λem = 530 nm) to yellow (λem = 561 nm) because of monosubstitution with glutathione (I561nm/I530nm). Addition of excess glutathione caused the second stage of ratiometric fluorescent color change from yellow to reddish orange (λem = 596 nm, I596nm/I561nm) due to disubstitution with glutathione. Therefore, different concentration ranges of glutathione (from less to excess) could be rapidly detected by the two-stage ratiometric fluorescent probe 1 in 5 min. While, probe 2 shows single-stage ratiometric fluorescent detection to GSH (from green to reddish orange, I596nm/I535nm). Probes 1 and 2 exhibit excellent properties with sensitive, specific colorimetric response and ratiometric fluorescent response to glutathione over other sulfur nucleophiles. Application to cellular ratiometric fluorescence imaging indicated that the probes were highly responsive to intracellular glutathione.

Foot-and-mouth disease virus-like particles as integrin-based drug delivery system achieve targeting anti-tumor efficacy.

The surface of foot-and-mouth disease virus (FMDV)-like particles (VLPs) contains a conserved arginine-glycine-aspartic acid (RGD) motif. Natural FMDV specifically attaches to overexpressed integrin receptors in several cancer cells. The FMDV VLPs produced in Escherichia coli were used for the first time as a delivery system of anti-tumor drug doxorubicin (DOX). The DOX-loaded VLPs exhibited a distinct release profile in different physiological conditions. The effects of FMDV-VLPs-DOX on cellular internalization and viability were evaluated in vitro by cell imaging, MTT assay and apoptosis, respectively. The anti-tumor efficacy in vivo was also determined in a nude mouse xenograft model based on tumor volume/weight and histological changes. The FMDV-VLPs-DOX complex significantly inhibited the proliferation of tumor and improved the pathological damage of DOX to non-targeting tissues. All results supported the potential of FMDV VLPs as a platform for specific targeted delivery of drugs or chemical reagents.

​The structural regulation of photosensitive unit and conjugation in COFs for efficient photocatalytic H2 evolution.

The photosensitive unit and conjugation play a significant role in photocatalytic performance of covalent organic frameworks (COFs). In this work, a series of COFs that introduced the phenyl phenanthridine as photosensitive unit with different planarity of linkages were synthesized and the common regulation between them for photocatalysis hydrogen evolution reaction (HER) was also studied. The results indicate that DHTB-PPD, with 2/3 planarity linkages (ß-ketoenamine/imine is 2/3) and the phenyl phenanthridine as building blocks, shows the narrowest bandgap and the strongest charge separation efficiency. Therefore, it shows the highest H2 production rate of 12.13 mmol g-1 h-1. The optimal photocatalytic efficiency of DTHB-PPD can be attributed to the combined effect of the photosensitive unit and the long-range ordering of the COF skeleton. According to The Density Functional Theory (DFT), the O site on ß-ketoamine is the most possible H2 generation site, but the photocatalytic efficiency of TP-PPD, with the highest skeletal conjugation and the highest proportion of ß-ketoamine is not the most efficient photocatalyst, indicating that the long-range ordering of COFs is important on photocatalytic performance. Thus, these findings provide valuable guidance for the structural design of COFs photocatalysts.

ß-galactosidase-activated red fluorescent probe assists in the diagnosis of pancreatitis.

Acute pancreatitis (AP) is an acute inflammatory disease resulting from abnormal digestion of itself and surrounding organs by pancreatic enzymes caused by a variety of pathogenic factors. When the tissue of a biological organism is afflicted with pancreatitis and experiences swelling, bleeding, and necrotic injuries, the abnormal expression of ß-galactosidase (ß-Gal) activity becomes one of the main indicators for assisting in the diagnosis of pancreatitis. In this study, a highly specific red fluorescent probe designed for the detection of ß-Gal activity has been developed. ß-galactoside is used as the enzyme activating group, and the long-wavelength luminescent water-soluble organic molecule NBDOH is used as the luminophore to construct the fluorescent probe NBD-gal. NBD-gal is activated by abnormally overexpressed ß-Gal, releasing a strong red fluorescent signal. The new fluorescent probe developed in this study is used to assist in the diagnosis of pancreatitis by detecting abnormal expression of ß-Gal activity in vivo. In in vivo imaging experiments, NBD-gal can effectively differentiate between normal nude mice and pancreatitis nude mice. In addition, NBD-gal can be effectively localized in real-time to the pancreas and intestines, which are rich in ß-Gal in nude mice. Therefore, NBD-gal exerts great potential in the early diagnosis, treatment, and prevention of biomedical clinical pancreatitis.

Photophysical study of a polyoxo ethylene linked naphthalene-based fluorescent chemosensor for Mg2+ and Ca2+ detection.

A naphthalene-based bichromophoric fluorescent sensor 2,2'-[oxy-bis(2-oxatetramethyleneoxy)]-bis[N-(2-naphthyl)-benzamide)] (1) was synthesized and characterized. Fluorescence decay for 1 in alcoholic solvents in the region of 415-460 nm revealed bi-exponential behavior. The faster component of the decay can be attributed to the formation of dimers. Above 480 nm, besides the dimer, there is also a little excimer formation and this excimer emits at longer wavelengths than the dimer. The observation of the change of the fluorescence emission spectra upon addition of water in EtOH-water mixtures is in line with the formation of water-bridged complexes preventing excimer formation. The sensor shows an increase in fluorescence intensity upon increasing Mg(2+) or Ca(2+) concentration in EtOH because the formation of the excimer can be hindered upon complexation with Mg(2+) or Ca(2+) ions. Because of the competition between hydrated metal ions and the water-bridged complex, spectral changes by complexation with Mg(2+) or Ca(2+) in EtOH-H2O (9 : 1 v/v) are quite different from those in neat ethanol. The ground-state dissociation constant K(d) estimated for the complex with Mg(2+) or Ca(2+) was found to be around 2.0 mM in EtOH-H2O (9 : 1 v/v), which makes it suitable for the measurement of the concentrations of these ions in physiologically relevant concentration ranges.

Covalent organic framework-MnO2 nanoparticle composites for shape-selective sensing of bithiols.

Covalent organic frameworks (COFs) for detecting biological macromolecules in water or biological environments are generally challenging. In this work, a composite material IEP-MnO2 is obtained by combining manganese dioxide (MnO2) nanocrystals and a fluorescent COF (IEP), which is synthesized by using 2,4,6-tris(4-aminophenyl)-s-triazine and 2,5-dimethoxyterephthalaldehyde. By the addition of biothiols, such as glutathione, cysteine or homocysteine with different sizes, the fluorescence emission spectra of IEP-MnO2 changed ("turn-on" or "turn-off") via different mechanisms. The fluorescence emission of IEP-MnO2 increased in the presence of GSH by the elimination of the FRET (Förster resonance energy transfer) effect between MnO2 and IEP. Surprisingly, due to the formation of a hydrogen bond between Cys/Hcy and IEP, the fluorescence quenching for IEP-MnO2 + Cys/Hcy may be explained via the photoelectron transfer (PET) process, which endows IEP-MnO2 with specificity in distinguishing the detection of GSH and Cys/Hcy compared to other MnO2 complex materials. Therefore, IEP-MnO2 was used to detect GSH and Cys in human whole blood and serum, respectively. The limit of detection for GSH in whole blood and Cys in human serum was calculated to be 25.58 µM and 4.43 µM, which indicates that IEP-MnO2 can be used to investigate some diseases related to GSH and Cys concentration. Moreover, the research expands the application of covalent organic frameworks in the fluorescence sensing field.

High-performance photocatalytic peroxymonosulfate activation by carbon quantum dots via precise surface chemistry regulation: Insight into the structure-function relations.

Carbon quantum dots (CQDs) are considered promising metal-free green catalysts for the activation of persulfates, but direct experimental evidence to identify the true active sites on the surface of CQDs is still lacking. We prepared CQDs with different oxygen contents by controlling the carbonisation temperature, using a simple pyrolysis method. Photocatalytic activity experiments show that CQDs200 exhibits the best PMS activation performance. By investigating the relationship between the oxygen functional groups on CQDs surface and photocatalytic activity, it was postulated that the C=O groups might be the predominant active site, which was confirmed by selective chemical titrations of the C=O, C-OH and COOH groups. Furthermore, limited to the weak photocatalytic properties of the pristine CQDs, ammonia and phenylhydrazine were used to precisely nitrogen-modify the o-CQD surface. We found that phenylhydrazine-modified o-CQDs-PH promoted the absorption of visible light and the separation of photocarriers, thus enhancing the activation of PMS. Theoretical calculations provide more insights from different levels of the pollutant, fine-tuned CQDs, and their interactions.

A self-assembled nanophotosensitizer targets lysosomes and induces lysosomal membrane permeabilization to enhance photodynamic therapy.

We report the self-assembly of amphiphilic BDQ photosensitizers into lysosome-targeting nanophotosensitizer BDQ-NP for highly effective photodynamic therapy (PDT). Molecular dynamics simulation, live cell imaging, and subcellular colocalization studies showed that BDQ strongly incorporated into lysosome lipid bilayers to cause continuous lysosomal membrane permeabilization. Upon light irradiation, the BDQ-NP generated a high level of reactive oxygen species to disrupt lysosomal and mitochondrial functions, leading to exceptionally high cytotoxicity. The intravenously injected BDQ-NP accumulated in tumours to achieve excellent PDT efficacy on subcutaneous colorectal and orthotopic breast tumor models without causing systemic toxicity. BDQ-NP-mediated PDT also prevented metastasis of breast tumors to the lungs. This work shows that self-assembled nanoparticles from amphiphilic and organelle-specific photosensitizers provide an excellent strategy to enhance PDT.

Visible absorption and fluorescence spectroscopy of conformationally constrained, annulated BODIPY dyes.

Six conformationally restricted BODIPY dyes with fused carbocycles were synthesized to study the effect of conformational mobility on their visible electronic absorption and fluorescence properties. The symmetrically disubstituted compounds (2, 6) have bathochromically shifted absorption and fluorescence spectral maxima compared to those of the respective asymmetrically monosubstituted dyes (1, 5). Fusion of conjugation extending rings to the α,ß-positions of the BODIPY core is an especially effective method for the construction of boron dipyrromethene dyes absorbing and emitting at longer wavelengths. The fluorescence quantum yields Φ of dyes 1-6 are high (0.7 ≤ Φ ≤ 1.0). The experimental results are backed up by quantum chemical calculations of the lowest electronic excitations in 1, 2, 5, 6, and corresponding dyes of related chemical structure but without conformational restriction. The effect of the molecular structure on the visible absorption and fluorescence emission properties of 1-6 has been examined as a function of solvent by means of the recent, generalized treatment of the solvent effect, proposed by Catalán (J. Phys. Chem. B 2009, 113, 5951-5960). Solvent polarizability is the primary factor responsible for the small solvent-dependent shifts of the visible absorption and fluorescence emission bands of these dyes.

A novel dual-capability naphthalimide-based fluorescent probe for Fe3+ ion detection and lysosomal tracking in living cells.

We design and synthesize a novel 1,8-naphthalimide-based fluorescent probe MNP that features the dual capabilities of tracking lysosomes in living HeLa cells and sensitively detecting Fe3+ ions in aqueous solution. The MNP is obtained by modifying the morpholine group with a lysosomal targeting function and the piperazine group with an Fe3+ ion recognition function on the 1,8-naphthalimide matrix. In the presence of Fe3+ ions, the MNP acts as a recognition ligand to coordinate with the central Fe3+ ion, and the protonated [MNPH]+ is eventually generated, in which significant fluorescence enhancements are observed due to the intramolecular photo-induced electron transfer (PET) process being blocked. The limit of detection of Fe3+ ions is as low as 65.2 nM. A cell imaging experiment shows that the MNP has low cytotoxicity and excellent lysosomal targeting ability. Therefore, the MNP offers a promising tool for lysosomal tracking and relevant life process research.

Near-infrared probe for early diagnosis of diabetic complications-nephropathy and in vivo visualization fluorescence imaging research.

Diabetic nephropathy is one of the common complications of diabetes, which has high risk of renal function. Dipeptidyl peptidase 4 (DPP4) is considered to be one of the good dynamic monitoring indicators for early diabetic nephropathy. Therefore, real-time monitoring of changes in the activity of DPP4 in organisms is helpful to the diagnosis and treatment of diabetes and its complications-diabetic nephropathy. A near-infrared fluorescent probe GP-DCMNH2 is designed to detect the activity of DPP4. GP-DCMNH2 is catalyzed and hydrolyzed by DPP4 into the near-infrared fluorescent dye DCMNH2, to achieve the purpose of detecting DPP4 in organisms. Based on the excellent near-infrared spectroscopy characteristics displayed by the probe GP-DCMNH2 in vitro, in living cells and diabetic mouse models, GP-DCMNH2 has been further applied in the visual fluorescence imaging of diabetic complications-diabetic nephropathy. Compared with the control group and the treatment group, GP-DCMNH2 showed a stronger near-infrared fluorescence signal in the kidney tissue and blood of diabetic nephropathy mice. Because GP-DCMNH2 shows high sensitivity in real-time dynamic monitoring of changes in the activity of DPP4 in organisms, and shows strong practicability in the spectral test of mouse models of diabetes and diabetic nephropathy. In clinical medicine, GP-DCMNH2 is expected to be used in the early diagnosis, prevention and treatment of diabetes and diabetic nephropathy.

Covalent Organic Frameworks Doped with Different Ratios of OMe/OH as Fluorescent and Colorimetric Sensors.

Improving the luminescence properties of covalent organic frameworks (COFs) has always been an important issue. Here, a series of COFs (([OMe]x -TzDa (TzDa is composed only by monomerics Tz and Da, OMe represents the incorporation of monomeric Dm)) with different ratios of OMe and OH were designed and synthesized. The photochemical behavior of [OMe]x -TzDa changed significantly due to the synergistic effect of aggregation induced emission (AIE), intramolecular charge transfer (ICT), and excited-state intramolecular proton transfer (ESIPT) effects. [OMe]2 -TzDa, which contained a ratio of 2/1 of OMe/OH, showed the strongest fluorescence emission in water and the best linear relationship for the detection of pH. Furthermore, [OMe]2 -TzDa was used to monitor HCl and NH3 gases and showed a color change, visible to the naked eye. Therefore, a "confidential pigment" was successfully made. Moreover, [OMe]2 -TzDa was also applied to detect N2 H4 . The work indicates the [OMe]2 -TzDa can serve as the first fluorescence sensor to detect pH, HCl and NH3 gases, which also shows a good response to N2 H4 .

Structural Engineering of Covalent Organic Frameworks Comprising Two Electron Acceptors Improves Photocatalytic Performance.

Covalent organic frameworks (COFs) have recently attracted much attention as potential photocatalysts for hydrogen production. The effective separation of photogenerated charges is a key objective to improve the photocatalytic activity of COFs. Here, four COFs were synthesized through the Schiff-base reaction to investigate whether the presence (simultaneous or not) of triazine and ketone as acceptors in COFs improved electron-hole separation efficiency. Evidence indicated that charge separation was more efficient when triazine and ketone were simultaneously present in the COF. The COF comprising two acceptors displayed the highest photocatalytic hydrogen production rate (31.43 µmol h-1 ; 41.2 and 3.4 times as large as those of the COFs containing only triazine or ketone, respectively). Moreover, the effect of the distance between the two acceptors on the electron-hole separation was investigated by changing the length of a bridging biphenyl ring. It turned out that the transport distance of a single phenyl group was more favorable for the catalytic reaction. This work affords insight and support for the design of efficient COF photocatalysts.