Mitochondria-targeted ruthenium(II) complexes for photodynamic therapy and GSH detection in living cells.

Photodynamic therapy is an emerging tumor therapy that kills tumor cells by activating reactive oxygen species (ROS) produced by photosensitizers. Mitochondria, as an important organelle, are the main generator of cellular ROS. Therefore, the development of photosensitizers capable of targeting mitochondria could significantly enhance the efficacy of photodynamic therapy. In this study, two novel ruthenium(II) complexes, Ru-1 and Ru-2, were designed and synthesized, both of which were functionalized with α,ß-unsaturated ketones for sensing of glutathione (GSH). The crystal structures of the two complexes were determined and they exhibited good recognition of GSH by off-on luminescence signals. The complex Ru-2 containing aromatic naphthalene can enter the cells and react with GSH to generate a strong luminescence signal that can be used to monitor intracellular GSH levels through imaging. Ru-2 also has an excellent mitochondrial localization ability with a Pearson's coefficient of 0.95, which demonstrates that it can efficiently target the mitochondria of tumor cells to enhance the effectiveness of photodynamic therapy as a photosensitizer.

Dual-channel luminescent Ir(III) complex for detection of GSH and Hcy/Cys in cells.

Glutathione (GSH), homocysteine (Hcy) and cysteine (Cys) play important roles in many physiological processes. However, due to their structural and functional similarities, it is still a challenge to develop a probe that can differentiate between GSH and Hcy/Cys simultaneously. In this work, a luminescent probe Ir-NBD was designed and synthesized, which emit weakly due to the presence of photo induced electron transfer (PET) interaction. When it reacted with the three biothiols, NBD dissociated and luminescence of Ir-OH was enhanced in the near-infrared (NIR) region due to the disappearance of the PET effect. On the other hand, the products obtained from the reaction of GSH with NBD were hardly luminescent, but the products from the reaction of Hcy/Cys with NBD could undergo an intramolecular rearrangement, resulting in an enhanced luminescence of the solution in the visible region. Ir-NBD enabled highly selective and sensitive detection of GSH and Cys/Hcy in a relatively short time (15 min). The two luminescent colors were clearly differentiated without spectral interference and the detection limit reached 1.32 µM (GSH), 0.42 µM (Hcy) and 0.51 µM (Cys), respectively. Ir-NBD also had low cytotoxicity, it realized the simultaneous detection of GSH and Hcy/Cys by dual-channel luminescence, and also provided ideas for the design of multifunctional luminescent probes.

Two near-infrared phosphorescent iridium(III) complexes for the detection of GSH and photodynamic therapy.

GSH is one of the most important reducing agents in biological systems. The depletion of GSH in the human body is linked to many diseases. Therefore, it is necessary to develop suitable and efficient probes for detecting GSH concentrations in real samples. In this work, we designed and synthesized two near-infrared emitting iridium(III) complex probes containing a novel ligand functionalized with an α,ß-unsaturated ketone for the rapid and sensitive detection of GSH. The molecular structure of Ir2 was determined by X-ray crystallography. Due to their large Stokes shift, long luminescence lifetime and NIR emission, these probes were successfully applied in the imaging of GSH in living cells. In addition, two iridium(III) complexes have strong singlet oxygen generation ability which can be used for photodynamic therapy (PDT) upon visible light irradiation. On the basis of these findings, our iridium(III) complexes may serve as GSH probes for HeLa cell imaging and as photosensitizers for PDT.

Aggregation-Induced Near-Infrared Emission and Electrochemiluminescence of an Iridium(III) Complex for Ampicillin Sodium Sensing.

A new iridium(III) complex was synthesized and characterized. Its photophysical properties and aggregation-induced emission and electrochemiluminescence in the near-infrared range were studied. The large conjugated cyclometallic ligand 1,2-phenylbenzoquinoline (pbq) was selected to form the Ir-C bond with the metal iridium(III) center and provide near-infrared emission of the complex. The auxiliary ligand 4,4'-diamino-2,2'-bipyridine (dabpy) can form hydrogen bonds, which was beneficial for the generation of aggregation-induced emission. The complex was aggregated into small spherical nanoparticles in 80% water and fascinating nanorings in 90% water. The sensing of ampicillin sodium (AMP) antibiotic by the iridium(III) complex were also investigated by photoluminescent and electrochemiluminescent methods. The complex showed a good selectivity toward AMP antibiotic compared to sodium phenylacetate and other eight antibiotics. The detection limits for AMP antibiotic was 0.76 µg/mL. This work provided a new strategy for the design of iridium(III) complex-based aggregation-induced emission and electrochemiluminescence probes for the sensing application.

Silver Carp (Hypophthalmichthys molitrix) Scale Collagen Peptides-1 (SCPs1) Inhibit Melanogenesis through Downregulation of the cAMP-CREB Signaling Pathway.

The mechanism of silver carp scale collagen peptides (SCPs1) on melanogenesis and its mechanism of action were examined in mouse melanoma cells (B16). The cell viability and effects of SCPs1 on intracellular tyrosinase (TYR) activity and melanin, reactive oxygen species (ROS), glutathione (GSH) and cyclic adenosine monophosphate (cAMP) content were examined. The regulatory mechanism of SCPs1 on the cAMP response element-binding protein (CREB) signaling pathway was analyzed. The cell viability of the SCPs1 group was >80% (0.01-1 mg/mL) and the inhibitory rate of SCPs1 on B16 cell melanin increased in a dose-dependent manner. The highest inhibitory rate of SCPs1 on melanin content reaching 80.24%. SCPs1 significantly increased the GSH content and decreased the tyrosinase activity, as well as the content of ROS and cAMP. Western blot analysis showed that SCPs1 significantly inhibited melanocortin-1 receptor (MC1R) expression and CREB phosphorylation in the cAMP-CREB signaling pathway, leading to downregulation of microphthalmia-associated transcription factor (MITF) and the expression of TYR, TYR-related protein-1 (TRP-1) and TRP-2. SCPs1 also inhibited the expression of MC1R, MITF, TYR, TRP-1 and TRP-2 at the transcriptional level. Taken together, SCPs1 inhibited melanin synthesis through the downregulation of the cAMP-CREB signaling pathway. Fish-derived collagen peptides could potentially be applied in skin whitening products.

Silver Carp (Hypophthalmichthys molitrix) Scales Collagen Peptides (SCPs): Preparation, Whitening Activity Screening and Characterization.

This study involves the preparation of scale collagen peptides (SCPs) with whitening activity from silver carp (Hypophthalmichthys molitrix) and their characterization and peptide sequence identification. In this article, scanning electron microscopy (SEM) was used to observe structure changes of sliver carp scales; enzymatic hydrolysis was optimized through protease screening and response surface optimization. The ultrafiltration was used to separate SCPs and the whitening activity was comprehensively evaluated using radical scavenging rate and tyrosinase-inhibiting activity, among others. An optimal component was characterized and identified using various modern spectral analysis techniques. The results showed that the surface of silver carp scales after decalcification was smooth and clear. The pepsin had the highest peptide yield and tyrosinase-inhibiting activity (90.01% and 82.25%, respectively). The optimal enzymatic hydrolysis conditions were an enzyme dosage of 16.1%, a solid-liquid ratio of 1:15.6 and a time of 4.9 h. The proportions of hydrophobic and basic amino acids in the peptide composition were 32.15% and 13.12%, respectively. Compared with SCPs2, SCPs1 (6096.68-9513.70 Da) showed better ·OH scavenging ability, tyrosinase-inhibiting activity and moisture absorption. SCPs1 was a macromolecular fragment of type I collagen with a triple helix structure, containing three peptide sequences with the potential for tyrosinase activity inhibition (AGPPGADGQTGQRGE, SGPAGIAGPAGPRGPAGPNGPPGKD and KRGSTGEQGSTGPLGMRGPRGAA). These results show that SCPs1 is a collagen peptide product with whitening potential.

Analysis of Water Distribution and Muscle Quality of Silver Carp (Hypophthalmichthys molitrix) Chunks Based on Electron-Beam Irradiation.

Electron-beam irradiation (EBI) is an efficient, safe, and nonthermal sterilization technique that is extensively used in food preservation research. Here we report the effects of different EBI doses (0, 4, 8 kGy) and preservation temperatures (room temperature [RT], 4 °C) on the muscle water distribution and muscle quality indices of silver carp chunks (SCCs). The highest entrapped water content was found in the 4-kGy-irradiated/4-°C-stored samples. The expressible moisture content (EMC) of the SCCs increased with increasing irradiation dose and was significantly lower in the RT group than in the 4 °C group. The irradiation dose and preservation temperature had no significant effect on the moisture content, whiteness value and protein content of SCCs (p > 0.05). When the irradiation dose reached 8 kGy, AV value, POV value and TVB value were significantly increased (p < 0.05). The myofibrillar protein content and actomyosin content of the SCCs in the 4 °C group was higher than that of the specimens in the RT group by 0.29−0.98 mg/mL (p < 0.05) and 36.21−296.58 µg/mL (p < 0.05), respectively. Overall, EBI treatment (4 kGy) and low-temperature preservation (4 °C) helped retain the muscle water content of the SCCs and preserve their quality, thereby endorsing the EBI treatment of silver carp products.

Antimicrobial activities and mechanism of sturgeon spermary protein extracts against Escherichia coli.

This study aimed to evaluate the antimicrobial activities and mechanism of sturgeon spermary protein extracts (SSPE) against Escherichia coli. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined. Cell structural change was analyzed using scanning electron microscopy-energy dispersive X-ray spectrometry and transmission electron microscope. Moreover, pH, zeta potential, membrane potential, intracellular ATP concentrations and the interaction of SSPE with genomic DNA were analyzed. Results showed that molecular weight of SSPE is 13.4 kDa, the content of basic amino acids is the highest, in which arginine accounts for 73.2%. The MIC and MBC of SSPE for E. coli were 0.05 and 5 mg/mL, respectively. After SSPE treatment, cell membrane permeability changes, zeta potential decrease and genomic DNA lysis occurred in E. coli, which indicated it exerted bacteriostatic effects either independently or simultaneously by destroying the cell membrane and genomic DNA. These findings indicated that SSPE has potential to be a natural antiseptic.

A sensitive photoluminescent sensor based on highly charged monoruthenium(II) complexes for dopamine detection.

A sensitive and selective photoluminescent sensor based on the highly charged monoruthenium(II) complex was designed to detect dopamine (DA) in aqueous samples. Two novel highly charged cationic ruthenium(II) complexes [Ru(bpy)2(bpy-N)]X4 (bpy = 2,2'-bipyridine, bpy-N = 4,4'-bis[N,N,N-triethyl-(methylamino)]-2,2'-bipyridine, X- = [PF6]- (1a) or Cl- (1b) and [Ru(bpy)(bpy-N)2]X6 (X- = [PF6]- (2a) or Cl-(2b)) can be assembled with anionic surfactant sodium dodecylbenzene sulfonate (SDBS), leading to an enhancement of photoluminescence intensity. Upon addition of DA to the system, the photoluminescence intensity of the assembled system was quenched due to the energy transfer effect. It exhibited a wide linear range (0.1-50 µM) and low detection limit (10 nM). The sensor demonstrated a high selectivity toward DA, especially in the presence of adrenaline (Adr) and norepinephrine (NE), whose structures are similar to DA in biological systems. With the merits of simple operation, obvious phenomenon and fast response speed, the sensor had a potential application prospect in human urine sample.

Amino group-driven distinguishing homocysteine from cysteine and glutathione in photoluminesecent signal of the iridium(III) complexes.

In this work, six iridium(III) complexes have been designed, synthesized and characterized. The molecular structures of complex 1 ([(pba)2Ir(bpy-2N(CH3)2)]PF6), 2 ([(pba)2Ir(bpy-2NH2)]PF6) and 3 ([(pba)2Ir(bpy-2CH3)]PF6) were determined by single crystal X-ray diffraction. Upon addition of Hcy (homocysteine) to the solution of complex 1, a luminescent variation from orange red to green was observed by the naked eye, corresponding to a large blue shift from 604 nm to 498 nm (~106 nm). While the emission intensity of complex 1 was almost no change after addition of other common amino acids including Cys (cysteine) and GSH (glutathione). The aldehyde group of complex 1 formed a new thiazinane/thiazolidine ring with Hcy/Cys confirmed by 1H NMR and high-resolution mass spectrometry. And the new product 1-Hcy had a higher quantum yield than 1-Cys. Theoretical calculations showed that the HOMO (highest occupied molecular orbital) of 1-Hcy was located on the newly formed six-membered thiazinane ring, which was different from the HOMO of 1-Cys. Compared with the other iridium(III) complexes, we can speculate that the large blue shift and enhancement of the emission intensity of the complex 1 were related to the strong electron donating ability of the modified amino groups on bipyridine ligand. This will provide an idea for the design of ratio-based luminescence probes for Hcy in future.

An Ir(III) complex capable of discriminating homocysteine from cysteine and glutathione with luminescent signal and imaging studies.

A new cationic Ir(III) complex with aldehyde and amino groups was synthesized and characterized. The Ir(III) complex has rich photophysical properties. The reaction of the aldehyde group in the Ir(III) complex with homocysteine (Hcy) afforded thiazinane derivatives which resulted in obvious changes in the luminescence spectra. After addition of Hcy to the Ir(III) complex containing 4,4'-diamino-2,2'-bipyridine, the luminescence intensity at ca. 580-610 nm decreased, and a new band at ca.490-520 nm appeared and enhanced strongly with a large blue shift of ca.90 nm, and the luminescent color changed from orange red to green. Based on this ratiometric probe, it can sensitively and selectively recognize Hcy by the ratio of emission intensity at two wavelengths to the concentrations of Hcy. While after addition of cysteine (Cys) or glutathione (GSH), the luminescence band showed a mild decrease in intensity with an unnoticeable shift. These different phenomena make it capable of discriminating homocysteine from cysteine and glutathione. The cytotoxicity and imaging of the complex were also studied in this work. The complex exhibited very low cytotoxicity on HeLa cells and showed sensitivity toward Hcy in living cells. These advantages provide it a good candidate for the application in the analytical and bioanalytical field.

Iridium(III) and gadolinium(III) loaded and peptide-modified silica nanoparticles for photoluminescence and magnetic resonance (dual) imaging.

The combination of non-invasive fluorescence (FL) and magnetic resonance (MR) imaging can compensate for disadvantages in terms of resolution and sensitivity. However, the preparation of dual-mode probes simultaneously exhibiting strong brightness and high MR response is challenging. A multifunctional nanoprobe was synthesized for targeted photoluminescence (PL) and MR dual-modal imaging. It was obtained by conjugating iridium(III) complexes, gadolinium(III) and the peptide arginine-glycine-aspartate (RGD) onto silica nanoparticles (Ir@SiO2-Gd-RGD NPs). They are highly water soluble, have an average diameter of ~50 nm, and emit strong yellowish green PL (with excitation/emission peaks at 380/572 nm). Simultaneously, the nanoprobes exhibit high MR response with a longitudinal relaxation of 7.16 mM-1 s-1. Instead of simple encapsulation, Ir(III) complexes were covalently conjugated to silica matrix to enhance the chemical and photochemical stability of the nanoprobes. The excellent biocompatibility and PL/MR dual modal imaging capability of the NPs is demonstrated using HeLa cells and mice as models.

The functionalized ruthenium(II) polypyridine complexes for the highly selective sensing of mercury ions.

A series of new ruthenium(II) polypyridine complexes appending with thioether groups were designed, synthesized and characterized. The sensing ability of the complexes toward mercury ions were studied by electronic absorption and emission spectra, and the reaction of the complexes with mercury ions were also confirmed by ESI mass spectroscopy and 1HNMR spectroscopy. The thioether groups would react with mercury ion fast to form aldehyde group leading to the significant change in the spectra. The color of the complex changed from yellow to orange after addition of mercury ions, and the color of the emission changed from red orange to dark red with a large red shift (~80 nm). Importantly, these kinds of ruthenium(II) complexes show a unique recognition of mercury ions over other metal ions. The complexes with more thioether groups also showed a better sensitivity toward mercury ions, this is good strategy for the further design of the new phosphorescent probes for sensing of mercury ions.

Sensitive determination of lysozyme by using a luminescent and colorimetric probe based on the aggregation of gold nanoparticles induced by an anionic ruthenate(II) complex.

The negatively charged ruthenate(II) complex [Ru(bpy)(PPh3)(CN)3]- and gold nanoparticles (AuNPs) were used for detecting lysozyme (LYS). The luminescence of the ruthenate(II) complex is quenched by AuNPs, and this induces the aggregation of AuNPs and a color change from red to blue. After addition of lysozyme, the positively charged lysozyme and the negatively charged ruthenate(II) complex bind each other by electrostatic interaction firstly. This prevents AuNPs from aggregation and quenches the emission of the ruthenate(II) complex. Its luminescence and the degree of aggregation of the AuNPs can be used to quantify LYS. The fluorometric calibration plot is linear in the 0.01 to 0.20 µM LYS concentration range, and the calibration plot is linear between 0.02 and 0.20 µM of LYS. The color of the solution can be easily distinguished by bare eyes at 0.08 µM or higher concentration of LYS. The applicability of the method was verified by the correct analysis of LYS in chicken egg white. Graphical abstract Schematic of a luminometric and colorimetric probe based on the induced aggregation of gold nanoparticles by an anionic luminescent ruthenate(II) complex or sensitive lysozyme detection.

Coupling coumarin to gold nanoparticles by DNA chains for sensitive detection of DNase I.

A kind of coumarin-modified gold nanoparticle by the bridge of dsDNA chains was designed and synthesized for sensitive detection of DNase I. The fluorescence of coumarin 343 at emission wavelengths of 491 nm excited at 440 nm was quenched by the gold nanoparticles due to the energy transfer process after the coumarin 343 was connected on the gold nanoparticles by DNA chains. When dsDNA chains were cut off by DNase I, the coumarin 343 molecules were released from gold nanoparticles and the fluorescence of coumarin 343 would be restored. The DNase I activity could be detected by this fluorescence assay with a high sensitivity based on the change of the energy transfer efficiency. The intensity of restored fluorescence is linearly related to the quantity of DNase I in the range from 1.0 to 40 mU/mL with a detection limit of 0.22 mU/mL. This design idea could render a useful way to develop similar molecular or enzyme sensor in analytical or biological fields.

Barbituric acid-modified graphitic carbon nitride nanosheets for ratiometric fluorescent detection of Cu2.

In this work, barbituric acid-modified graphitic carbon nitride (BCN) nanosheets with blue-green fluorescence were prepared by a copolymerization of dicyandiamide with barbituric acid and then by a chemical oxidation process. A ratiometric fluorescent sensor for Cu2+ was designed by using BCN nanosheets as the report probe and Ru(bpy)3Cl2 as the reference probe. In the presence of different concentrations of Cu2+, the blue-green fluorescence of BCN nanosheets was drastically quenched, while the red fluorescence of Ru(bpy)3Cl2 remained constant. The color of the sensor changed from blue-green to red, resulting in ratiometric and visual detection of Cu2+ ions. The detection limit for Cu2+ was estimated to be 70 nM, and as low as 5 µM Cu2+ was distinguished with the naked eye. Additionally, this sensor was also applied for the detection of Cu2+ in tap water samples.

A luminescent and colorimetric probe based on the functionalization of gold nanoparticles by ruthenium(ii) complexes for heparin detection.

A sensitive assay was demonstrated for the colorimetric and luminescent detection of heparin. Positively charged [Ru(phen)2np]2+ (np: naphtho[2,3-f][1,10]phenanthroline) can interact with negatively charged AuNPs through electrostatic interaction, leading to the luminescence quenching of [Ru(phen)2np]2+via energy transfer as well as the aggregation of AuNPs together with a color change in solution from red to blue. Upon the addition of negatively charged heparin to gold nanoparticles functionalized by ruthenium(ii) complexes, the emission of [Ru(phen)2np]2+ recovered due to the competitive adsorption of [Ru(phen)2np]2+ to heparin and AuNPs. A linear calibration curve was obtained in the 0.4 µM to 100 µM range and 0.1 µM to 6 µM with a low detection limit of 0.22 µM and 0.024 µM based on 3σ/s for heparin quantification in buffer solution and in 1% fetal bovine serum (FBS), respectively. Interestingly, the naked eye alone can judge the presence of 0.02 µM heparin without the aid of any advanced instruments by color change. Thus a convenient and visual method to detect heparin was established.

Gold Nanoparticle-Based Probe for Colorimetric Detection of Copper Ions.

A new ligand 10-mercaptodecyl-1-iminodiacetic acid (MDIA) was synthesized and used to modify gold nanoparticles to provide a simple assay to repeatedly sense Cu²âº in the solution at room temperature. This functionalized gold nanosensor was applied for the detection of Cu²âº in water samples with sensitivity and simplicity. The chelation/aggregation process is reversible via addition of a strong metal ion chelator such as EDTA. This simple and fast colorimetric sensor is important in the application of copper ion detection in water quality during the emergency and early warning monitoring.

Colorimetric and luminescent bifunctional iridium(III) complexes for the sensitive recognition of cyanide ions.

Two new cyclometalated iridium(III) complexes [(ppy)2Irppz]Cl (1) and [(ppy)2Irbppz]Cl (2) (where ppy=2-phenylpyridine, ppz=4,7-phenanthrolino-5,6:5,6-pyrazine, bppz=2.3-di-2-pyridylpyrazine), were designed and synthesized. The structure of [(ppy)2Irppz]Cl was determined by single crystal X-ray diffraction. Their photophysical properties were also studied. This kind of complexes could coordinate with Cu2+, the photoluminescence (PL) of the complex was quenched, and the color changed from orange-red to green. The forming M-Cu (M: complexes 1 and 2) ensemble could be further utilized as a colorimetric and emission "turn-on" bifunctional detection for CN-, especially for complex 1-Cu2+ showed a high sensitivity toward CN- with a limit of diction is 97nM. Importantly, this kind of iridium(III) complexes shows a unique recognition of cyanide ions over other anions which makes it an eligible sensing probe for cyanide ions.