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Long-wavelength fluorescent proteins (LWFPs) and LWFP-based sensors are indispensable tools for bioimaging and biosensing applications. However, it remains challenging to develop LWFPs with outstanding brightness and/or sensitivities, largely due to the lack of simple and effective molecular design strategies. Herein, we rationalized the molecular origins of a multi-donor strategy that affords significant bathochromic shifts and large Stokes shifts with minimal structural changes in the resulting protein fluorophores. We analyzed three key factors that affect the spectral properties of these fluorophores, including the (1) substituent position, (2) electron-donating strength, and (3) number of electron-donating groups. We further demonstrated that this simple design strategy is generalizable to various fluorophore families. We expect that this work can provide rational guidelines for developing fluorescent proteins (and small-molecule fluorophores) with long emission wavelengths and large Stokes shifts.
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Electrones , Colorantes Fluorescentes , Colorantes Fluorescentes/química , HumanosRESUMEN
Long-term super-resolution imaging appears to be increasingly important for unraveling organelle dynamics at the nanoscale, but is challenging due to the need for highly photostable and environment-sensitive fluorescent probes. Here, we report a self-blinking fluorophore that achieved 12â nm spatial resolution and 20â ms time resolution under acidic lysosomal conditions. This fluorophore was successfully applied in super-resolution imaging of lysosomal dynamics over 40â min. The pH dependence of the dye during blinking made the fluorophore sensitive to lysosomal pH. This probe enables simultaneous dynamic and pH recognition of all lysosomes in the entire cell at the single-lysosome-resolved level, which allowed us to resolve whole-cell lysosome subpopulations based on lysosomal distribution, size, and luminal pH. We also observed a variety of lysosome movement trajectories and different types of interactions modes between lysosomes.
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Parpadeo , Colorantes Fluorescentes , Colorantes Fluorescentes/metabolismo , Células HeLa , Humanos , Concentración de Iones de Hidrógeno , Lisosomas/metabolismoRESUMEN
Food flavour is important in appetite control. The effects of aroma and taste, independently or in combination, on appetite sensation and subsequent food intake, were studied. Twenty-six females (24 ± 4 years, 20.9 ± 1.9 kgâ m-2) consumed, over 15 min period, one of four sample drinks as a preload, followed by an ad libitum consumption of a pasta meal (after 65 min). Sample drinks were: water (S1, 0 kcal), water with strawberry aroma (S2, 0 kcal), water with sucrose and citric acid (S3, 48 kcal) and water with strawberry aroma, sucrose and citric acid (S4, 48 kcal). Appetite sensation did not differ between the S1 (water), S2 (aroma) and S3 (taste) conditions. Compared with S1 (water), S2 (aroma) and S3 (taste), S4 (aroma + taste) suppressed hunger sensation over the 15 min sample drink consumption period (satiation) (p < 0.05). S4 (aroma + taste) further reduced hunger sensation (satiety) more than S1 at 5, 20 and 30 min after the drink was consumed (p < 0.05), more than S2 (aroma) at 5 and 20 min after the drink was consumed (p < 0.05), and more than S3 (taste) at 5 min after the drink was consumed (p < 0.05). Subsequent pasta energy intake did not vary between the sample drink conditions. S4 (aroma + taste) had the strongest perceived flavour. This study suggests that the combination of aroma and taste induced greater satiation and short-term satiety than the independent aroma or taste and water, potentially via increasing the perceived flavour intensity or by enhancing the perceived flavour quality and complexity as a result of aroma-taste cross-modal perception.
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Regulación del Apetito , Bebidas , Ingestión de Energía , Calidad de los Alimentos , Odorantes , Respuesta de Saciedad , Gusto , Adolescente , Adulto , Estudios Cruzados , Sacarosa en la Dieta/metabolismo , Inglaterra , Femenino , Preferencias Alimentarias , Humanos , Almuerzo , Investigación Cualitativa , Adulto JovenRESUMEN
The emission intensities of coumarin 545 solution exhibit a low temperature dependence, with a record-low temperature coefficient of only â¼0.025% per °C. This monomer-aggregate coupled fluorescence system can be used for ratiometric temperature measurements with high spatial and temporal resolutions; three different working modes have been demonstrated.
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Cumarinas/química , Colorantes Fluorescentes/química , Frío , Espectrometría de Fluorescencia , Temperatura , TermómetrosRESUMEN
The effective delivery and targeted release of drugs within tumor cells are critical factors in determining the therapeutic efficacy of nanomedicine. To achieve this objective, a conjugate of maltose (Mal) and bovine serum albumin (BSA) was synthesized by the Maillard reaction and self-assembled into nanoparticles with active-targeting capabilities upon pH/heating induction. This nanoparticle could be effectively loaded with doxorubicin (DOX) to form stable nanodrugs (Mal-BSA/DOX) that were sensitive to low pH or high glutathione (GSH), thereby achieving a rapid drug release (96.82 % within 24 h). In vitro cell experiments indicated that maltose-modified BSA particles efficiently enhance cellular internalization via glucose transporters (GLUT)-mediated endocytosis, resulting in increased intracellular DOX levels and heightened expression of γ-H2AX. Consequently, these results ultimately lead to selective tumor cells death, as evidenced by an IC50 value of 3.83 µg/mL in HepG2 cells compared to 5.87 µg/mL in 293t cells. The efficacy of Mal-BSA/DOX in tumor targeting therapy has been further confirmed by in vivo studies, as it effectively delivered a higher concentration of DOX to tumor tissue. This targeted delivery approach not only reduces the systemic toxicity of DOX but also effectively inhibits tumor growth (TGI, 75.95 %). These findings contribute valuable insights into the advancement of targeting-albumin nanomedicine and further support its potential in tumor treatment.
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Neoplasias Hepáticas , Nanopartículas , Humanos , Maltosa , Portadores de Fármacos , Doxorrubicina/farmacología , Sistemas de Liberación de Medicamentos , Albúmina Sérica Bovina , Neoplasias Hepáticas/tratamiento farmacológico , Glutatión , Concentración de Iones de HidrógenoRESUMEN
The formation mechanism behind the sophisticated aromas of sesame oil (SO) has not been elucidated. The interaction effects of the Maillard reaction (MR) and lipid oxidation on the aroma formation of fragrant sesame oil were investigated in model reaction systems made of l-lysine (Lys) and d-glucose (Glc) with or without fresh SO (FSO) or oxidized SO (OSO). The addition of OSO to the Lys-Glc model increased the MR browning at 294 nm and 420 nm and enhanced the DPPH radical scavenging activity greater than the addition of FSO (p < 0.05). The presence of lysine and glucose inhibited the oxidation of sesame oil, reduced the loss of γ-tocopherol, and facilitated the formation of sesamol (p < 0.05). The Maillard-lipid interaction led to the increased concentrations of some of the alkylpyrazines, alkylfurans, and MR-derived ketones and acids (p < 0.05) while reducing the concentrations of other pyrazines, lipid-derived furans, aliphatic aldehydes, ketones, alcohols, and acids (p < 0.05). The addition of FSO to the MR model enhanced the characteristic roasted, nutty, sweet, and fatty aromas in sesame oil (p < 0.05), while excessive lipid oxidation (OSO) brought about an unpleasant oxidized odor and reduced the characteristic aromas. This study helps to understand the sophisticated aroma formation mechanism in sesame oil and provides scientific instruction for precise flavor control in the production of sesame oil.
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Glucosa , Lisina , Reacción de Maillard , Odorantes , Oxidación-Reducción , Aceite de Sésamo , Aceite de Sésamo/química , Glucosa/química , Odorantes/análisis , Lisina/química , Fenoles/química , BenzodioxolesRESUMEN
The study characterized the aroma-active compounds produced by sesame hulls at three roasting temperatures and analyzed the similarities and differences in the aroma profile of sesame hulls with whole seeds and kernels after roasting. Roasting hulls produced mainly furans, aldehydes, and ketones volatiles. 140 Compounds were identified as aroma-active compounds, including 36 key aroma compounds (odor activity value, OAV ≥ 1). Among them, furanone (caramel-like, OAV = 80), 3-methylbutanal (fruity, OAV = 124), and 2-methoxy-4-vinylphenol (burnt, smoky, OAV = 160) gave hulls (180 °C) sweet, burnt, and smoky aroma. Due to the contribution of vanillin (fatty, sweet milk, OAV = 45), 2-hydroxy-3-butanone (caramel-like, roast, OAV = 46), and 2-methoxy-4-vinylphenol (OAV = 78), hulls (200 °C) shown strong sweet and roast note. These results identified compounds that contributed significantly to the aroma of sesame hulls and elucidated the contribution of sesame hulls to the flavor of roasted whole seeds and sesame oil.
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Two selenized chitooligosaccharide (O-Se-COS and N,O-Se-COS) with different sites modification were synthesized to alleviate liver injury in vivo. Comparing to traditional COS, both selenized COS exhibited enhanced reducibility as well as antioxidant capacity in vitro. Furthermore, O-Se-COS demonstrated superior efficacy in reducing intracellular reactive oxygen species (ROS) and mitochondrial damage compared to N,O-Se-COS as its enhanced cellular uptake by the positive/negative charge interactions. Two mechanisms were proposed to explained these results: one is to enhance the enzymatic activity of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px), which effectively scavenge free radicals; the other is to down-regulate intracellular cytochrome P450 (CYP2E1) levels, inhibiting carbon tetrachloride (CCl4)-induced peroxidation damage. In vivo studies further demonstrated the effective alleviation of CCl4-induced liver injury by selenized COS, with therapeutic efficacy observed in the following order: O-Se-COS > N,O-Se-COS > COS. Finally, hemolysis and histological tests confirmed the biosafety of both selenized COS. Taken together, these finding demonstrated that selenium has the potential to improve the biological activity of COS, and precise selenylation was more conducive to achieving the synergistic effect where 1 + 1>2.
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Quitosano , Hígado , Oligosacáridos , Selenio , Antioxidantes/farmacología , Tetracloruro de Carbono/toxicidad , Tetracloruro de Carbono/metabolismo , Especies Reactivas de Oxígeno/metabolismo , Quitina/farmacología , Quitina/uso terapéutico , Quitina/metabolismo , Estrés Oxidativo , Selenio/farmacología , Selenio/metabolismoRESUMEN
The unknown effect of sesame lignans on aroma formation in sesame oil via the Maillard reaction (MR) and lipid oxidation was investigated. Sesamin, sesamolin, or sesamol was added to 3 models: lysine+glucose (MR), cold-pressed sesame oil (SO), and MR + SO, and were heated at 120 °C for 60 min. All three lignans suppressed SO oxidation while increasing DPPH scavenging ability (p < 0.05). Lignans increased depletions of lysine and glucose and MR browning (p < 0.05). Lignans reduced most aroma-active pyrazines, aldehydes, ketones, alcohols, and esters (p < 0.05). Sesamol and sesamolin increased perceptions of the preferable aromas of nutty, roasted sesame, and popcorn while reducing the undesirable green and rancid aromas (p < 0.05). Sesamol demonstrated a stronger effect on lipid oxidation, MR browning, aroma formation, and sensory perception than sesamin and sesamolin. This study suggests that sesame lignans can modulate aroma formation and sensory perception of sesame oil by interacting with the MR and lipid oxidation pathways.
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Lignanos , Reacción de Maillard , Odorantes , Oxidación-Reducción , Aceite de Sésamo , Sesamum , Lignanos/química , Aceite de Sésamo/química , Sesamum/química , Odorantes/análisis , Humanos , Fenoles/química , Dioxoles/química , Benzodioxoles/químicaRESUMEN
This study developed a novel and green method to produce fragrant sesame oil using microwaves and subcritical extraction (SBE). Sesame seeds were microwaved at 540 W for 0-9 min before subcritical propane extraction at 40 °C and 0.5 MPa. SBE caused less deformation to the cellular microstructure of sesame cotyledons while dramatically improving oil yield (96.7-97.1 %) compared to screw processing (SP) (53.1-58.6 %). SBE improved extraction rates for γ-tocopherol (381.1-454.9 µg/g) and sesame lignans (917.9-970.4 mg/100 g) in sesame oil compared to SP (360.1-443.8 µg/g and 872.8-916.8 mg/100 g, respectively). Microwaves generated aroma-active heterocyclics and phenolics faster than hot-air roasting in sesame oil with a better sensory profile. SBE had a higher extraction rate for aroma-active terpenes, alcohols, and esters while reducing the concentrations of carcinogenic PAHs and HCAs in sesame oil. The novel combination process of microwaves and subcritical extraction is promising in producing fragrant sesame oil with superior qualities.
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Achieving practical full projector compensation requires the projection display to adapt quickly to textured projection surfaces and unexpected movements without interrupting the display procedure. A possible solution to achieve this involves using a projector and an RGB camera and correcting both color and geometry by directly capturing and analyzing the projected natural image content, without the need for additional patterns. In this study, we approach full projector compensation as a numerical optimization problem and present a physics-based framework that can handle both geometric calibration and radiometric compensation for a Projector-camera system (Procams), using only a few sampling natural images. Within the framework, we decouple and estimate the Procams' factors, such as the response function of the projector, the correspondence between the projector and camera, and the reflectance of projection surfaces. This approach provides an interpretable and flexible solution to adapt to the changes in geometry and reflectance caused by movements. Benefitting from the physics-based scheme, our method guarantees both accurate color calculation and efficient movement and reflectance estimation. Our experimental results demonstrate that our method surpasses other state-of-the-art end-to-end full projector compensation methods, with superior image quality, reduced computational time, lower memory consumption, greater geometric accuracy, and a more compact network architecture. The data and source code are accessible at https://github.com/kylin-leo/FullProjectorCompensation.
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OBJECTIVES: This study aims to investigate the correlation between serum Galectin-3 levels and the risk of periodontitis in patients with type 2 diabetes mellitus (T2DM). METHODS: A total of 140 patients with T2DM admitted to the endocrinology department of Weifang People's Hospital, Affiliated to Weifang Medical College from July 2021 to November 2022 were selected and divided into T2DM without periodontitis group (T2DM group, n=67) and T2DM with periodontitis group (T2DMP group, n=73) according to whether they were combined with periodontitis. In the same period, 65 non-periodontitis volunteers with normal blood glucose were selected as healthy control group (NC group). Blood samples of all subjects were collected, and serum Galectin-3 levels and related laboratory indices were detected and compared among the three groups. RESULTS: Serum Galectin-3 levels in the NC, T2DM, and T2DMP groups were 3.81 (3.49, 4.15), 4.82 (4.25, 5.26), and 6.83 (5.19, 7.28) ng/mL, respectively. After adjusting for the influence of baseline data by multiple linear regression, serum Galectin-3 levels in the T2DMP and T2DM groups were significantly higher than those in the NC group (all P<0.05). Multiple linear regression analysis showed that serum Galectin-3 levels were positively correlated with homeostatic model of the insulin resistance index (ß=0.254, 95%CI:0.089-0.419, P=0.003), glycosylated hemoglobin A1c (ß=0.397, 95%CI: 0.049-0.745, P=0.026), and clinical attachment loss (ß=0.298, 95%CI: 0.024-0.572, P=0.033). After adjusting for the effects of covariates, binary logistic regression showed that serum Galectin-3 levels were significantly associated with the risk of periodontitis in patients with T2DM (OR=2.146, 95%CI: 1.260-3.655, P=0.005). Trend test showed that the risk of periodontitis in patients with T2DM increased with increasing serum Galectin-3 levels (Ptrend=0.011). Receiver operating characteristic (ROC) curve analysis showed that the area under the curve (AUC) of serum Galectin-3 in predicting T2DM periodontitis was 0.861 (95%CI: 0.801-0.920, Z=11.806, P<0.001). CONCLUSIONS: Serum Galectin-3 levels were elevated in patients with T2DM and pe-riodontitis and associated with the risk of periodontitis.
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Diabetes Mellitus Tipo 2 , Periodontitis , Humanos , Diabetes Mellitus Tipo 2/complicaciones , Galectina 3 , Periodontitis/complicaciones , Hemoglobina GlucadaRESUMEN
The unclear effects of microwaves, as a greener alternative to hot air, on sensory perception, aroma, and hazardous components of sesame oil were investigated. Microwaves (900 W, 6-10 min) created more seed porosity and cell destruction and facilitated more γ-tocopherol release in sesame oil (349.30-408.50 mg/kg) than 200 °C, 20 min hot air (304.90 mg/kg). Microwaves (6-10 min) generated more aromatic heterocyclics (42.40-125.12 mg/kg) and aldehydes (5.15-2.08 mg/kg) in sesame oil than hot air (25.59 mg/kg and 1.34 mg/kg). Microwaves (6 min) produced sesame oil with a stronger roasted sesame flavour, and weaker bitter and burnt flavour than hot air. Microwaves reduced harman (≤775.19 ng/g), norharman (≤1,069.99 ng/g), and benzo(a)pyrene (≤1.59 µg/kg) in sesame oil than hot air (1,319.85 ng/g, 1,168.40 ng/g, and 1.83 µg/kg). Appropriate microwave is a promising alternative to hot air in producing sesame oil with a better sensory profile, more bioactive, and less carcinogenic components.
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This study investigated the changes in aroma composition and perception of sunflower oils induced by seed roasting using sensory-oriented flavor analysis. Volatile compounds were extracted by solvent-assisted flavor evaporation and headspace solid-phase microextraction. Odorants were characterized by gas chromatography-olfactometry-mass spectrometry and aroma extract dilution analysis. The cold-pressed and roasted sunflower oils contained 13 and 50 odorants, respectively, with the flavor dilution factors between 1 and 256. Fifty-six odorants were newly identified in sunflower oils. Quantification of 26 important odorants by the external standard method revealed apparent changes induced by seed roasting in loss of terpenes, formation of Maillard reaction products, and the increase in lipid oxidation products. The most important odorants (odor active values, OAVs = 1-1857) in the cold-pressed sunflower oil included α-pinene (11,145 µg/kg), ß-pinene (4068 µg/kg), linalool (56 µg/kg), hexanal (541 µg/kg), octanal (125 µg/kg), α-phellandrene (36 µg/kg), and (E)-2-octenal (69 µg/kg), contributing to the raw sunflower seed, woody, green, earthy, and sweet aromas of the oil. The most important contributors (OAVs = 1-884) to the roasted, smoky, and burnt aromas of the roasted sunflower oil were 2- and 3-methylbutanal (6726 and 714 µg/kg), 2,6-dimethylpyrazine (2329 µg/kg), 2,5-dimethylpyrazine (12,228 µg/kg), 2,3-dimethylpyrazine (238 µg/kg), 2,3-pentanedione (1456 µg/kg), 2-pentylfuran (1332 µg/kg), 2,3-dimethyl-5-ethylpyrazine (213 µg/kg), and 1-pentanol (693 µg/kg). Aroma recombination of the key odorants in odorless sunflower oil adequately mimicked the general aroma profiles of sunflower oils. This study provides an important foundation for understanding the relationship between oil processing and aroma molecules of sunflower oils. PRACTICAL APPLICATION: The clear changes observed in the composition and concentrations of key aroma compounds explained the changes in sensory characteristics of sunflower seed oils induced by seed roasting on a molecular basis. Characterizing the key aroma-active composition of sunflower oil and investigating its relationship with oil processing could provide important practical applications for the sunflower oil industry in flavor regulation, quality control, product development, and process optimization.
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Helianthus , Compuestos Orgánicos Volátiles , Odorantes , Aceites , Olfatometría , Aceite de GirasolRESUMEN
This was the first study to compare the key aroma-active compounds that contributed to the different aroma profiles between roasted and cold-pressed sesame oils. Aroma compounds were extracted by headspace solid-phase micro-extraction (HS-SPME) and simultaneous distillation extraction (SDE) and were analysed using gas chromatography-olfactometry-mass spectrometry (GC-O-MS) and aroma extract dilution analysis (AEDA). The numbers of aroma-active compounds with the flavour dilution (FD) factors between 1 and 2048 were 57 and 16 in the roasted and cold-pressed sesame oils, respectively. A total of 28 volatile compounds were identified as aroma-active compounds in sesame oils for the first time. Important aroma compounds (FD ≥ 8) were quantified by the external standard method, and their odour activity values (OAV) were calculated as the ratio of their concentrations to odour thresholds in oil. The numbers of key aroma-active compounds defined by OAVs ≥ 1 were 23 (OAVs = 1-385) and 8 (OAVs = 1-42), respectively, in the roasted and cold-pressed sesame oils. 2-Methoxy-4-vinylphenol (smoked, 1924 µg/kg, OAV = 385), 2-methoxyphenol (smoked, 1488 µg/kg, OAV = 114) and pyrazines (roasted and nutty, 578-22750 µg/kg, OAV = 1-67) were the most important aroma-active compounds in the roasted sesame oil, whereas hexanal (green and fruity, 3094 µg/kg, OAV = 42) was the most important aroma-active compound in the cold-pressed sesame oil, followed by (E,E)-2,4-decadienal (earthy, 4170 µg/kg, OAV = 31), dimethyl sulfone (sulphur-like, 406 µg/kg, OAV = 20) and octanal (green and fruity, 901 µg/kg, OAV = 16). This study provides valuable information for manufacturers to achieve precise flavour control of sesame oil products.
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Odorantes , Aceite de Sésamo , Aromatizantes , Cromatografía de Gases y Espectrometría de Masas , Odorantes/análisis , OlfatometríaRESUMEN
Si-rhodamine has been extensively used in super-resolution fluorescence imaging in recent years. Its equilibrium between ring-closed nonfluorescent spirolactones and ring-opened fluorescent zwitterions endows Si-rhodamine with excellent fluorogenicity, membrane permeability, and photostability. In this paper, the equilibrium of Si-rhodamine between lactones and zwitterions was revealed to be greatly affected by various environmental factors, including molecular aggregation, solvent polarity, pH, metal ions, irradiation, and temperature. These environmental sensitivities make Si-rhodamine useful as a hydrochromic material, a fluorescent sensor array for metal ions or solvents, and a photoactivatable switch. Importantly, these results indicate that using Si-rhodamine as a fluorogenic probe or a blinking fluorophore in single-molecule localization super-resolution microscopy requires caution on possible false signals caused by its environmental sensitivity.
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The visualization of the microcosmic behavior of proteins in vivo is the key to real-time monitoring of proteins. A series of wash-free SNAP-tag probes were designed and synthesized based on the combination of SNAP-tag and small organic molecule fluorescent dyes. SNAP-tag, which specifically recognized O6-benzylguanine, could be labeled with a fluorophore (e. g., 1,8-naphthalimide) through the formation of covalent bonds. Furthermore, the change from a hydrophilic environment to the hydrophobic cavum of SNAP-tag realized a 2-13-fold enhancement in fluorescence. Through the fusion of SNAP-tag and the target protein, the probes could recognize the mitochondrial proteins (e. g., cytochrome oxidase, Cox8A) and nuclear proteins (e. g., H2B) in living cells. Besides, the fluorescent probes allowed the in-situ real-time monitoring of proteins without washing.
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Colorantes Fluorescentes , Proteínas/química , FluorescenciaRESUMEN
A fluorescent sensor for halogenated solvents termed AMN is reported. AMN shows strong fluorescence in most halogenated solvents (QE > 0.1) but weak fluorescence (QE<0.01) in most non-halogenated solvents. In chlorinated solvents, the fluorescence intensity decreased with the reduction of chlorine content. On the contrary, in brominated solvents the fluorescence intensity increased with the reduction of bromine content. It is worth mentioning that AMN displayed fluorescence emission centered at 520 nm in CCl4 with a quantum yield of 0.607, at 556 nm in CHCl3 with a quantum yield of 0.318, at 584 nm in CH2Cl2 with a quantum yield of 0.128, whereas in CHBr3 was centered at 441 nm with a quantum yield of 0.012. AMN was shown to have the ability to differentiate CCl4, CHCl3, CH2Cl2 and CHBr3 halogenated solvents.
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The fluorescence intensity of N,N-dimethyl-4-((2-methylquinolin-6-yl)ethynyl)aniline exhibits an unusual intensification with increasing temperature, by activating more vibrational bands and leading to stronger TICT emissions upon heating in dimethyl sulfoxide. Based on the different temperature dependence at various wavelengths, as shown in the TICT fluorescence spectrum, this dye can be employed to ratiometrically detect temperature.