A novel "Turn-Off-On" fluorescent probe for specific sequential detection of Cu2+ and glyphosate and its application in biological imaging.

As common pollutants, Cu2+ and glyphosate pose a serious threat to human health and the ecosystem. Herein, a fluorescent probe (E)-7-(diethylamino)-N'(4-(diethylamino)-2-hydroxybenzyl)-2-oxo-2H chromophore-3-carbazide (DDHC) was designed and synthesised for the sequential recognition of Cu2+ and glyphosate. DDHC has the advantages of a short synthesis path, easy-to-obtain raw materials, good anti-interference ability, and strong stability. The interaction of the DDHC-Cu2+ complexes with glyphosate allows the amino and carboxyl groups in glyphosate molecules to coordinate with Cu2+ strongly, competing for the Cu2+ in the DDHC-Cu2+ complexes and releasing the DDHC, leading to the recovery of fluorescence. The recognition was further validated through Job's plot, HRMS, and DFT calculations. In addition, the successful recovery of Cu2+ and glyphosate in different environmental water samples fully demonstrates the practical application potential of DDHC. Especially, DDHC has low cytotoxicity and can enter zebrafish and HeLa cells, rapidly reacting with Cu2+ and glyphosate in the body, generating visible fluorescence quenching and recovery phenomena, achieving real-time visual monitoring of exogenous Cu2+ and glyphosate in zebrafish and HeLa cells. The targeting and dual selectivity of DDHC greatly enhance its potential application value in the field of detection, providing important theoretical support for studying the fate of multiple pollutants in the environment.

Influence of para-substituted benzaldehyde derivatives with different push/pull electron strength groups on the conformation of human serum albumin and toxicological effects in zebrafish.

Excessive intake of benzaldehyde and its derivatives can cause irreversible damage to living organisms. Hence, benzaldehyde derivatives with different para-substitutions of push/pull electronic groups were chosen to investigate the effect of different substituent properties on the structure of human serum albumin (HSA). The binding constants, number of binding sites, major interaction forces, protein structural changes, and binding sites of benzaldehyde (BzH) and its derivatives (4-BzHD) with HSA in serum proteins were obtained based on multispectral and molecular docking techniques. The mechanism of BzH/4-BzHD interaction on HSA is mainly static quenching and is accompanied by the formation of a ground state complex. BzH/4-BzHD is bound to HSA in a 1:1 stoichiometric ratio. The interaction forces for the binding of BzH/4-BzHD to HSA are mainly hydrogen bonding and hydrophobic interaction, which are also accompanied by a small amount of electrostatic interactions. The effect of BzH/4-BzHD on HSA conformation follows: 4-Diethylaminobenzaldehyde (4-DBzH) > 4-Nitrobenzaldehyde (4-NBzH) > 4-Hydroxybenzaldehyde (4-HBzH) > 4-Acetaminobenzaldehyde (4-ABzH) > BzH, which means that the stronger push/pull electronic strength of the para-substituted benzaldehyde derivatives has a greater effect on HSA conformation. Furthermore, the concentration-lethality curves of different concentrations for BzH/4-BzHD on zebrafish verified above conclusion. This work provides a scientific basis for the risk assessment of benzaldehyde and its derivatives to the ecological environment and human health and for the environmental toxicological studies of benzaldehyde derivatives with different strengths of push/pull electron substitution.

Enantioselective effects of chiral prothioconazole and its metabolites: Oxidative stress in HepG2 cells and lysozyme activity.

Chiral pesticides may exhibit enantioselectivity in terms of bioconcentration, environmental fate, and reproductive toxicity. Here, chiral prothioconazole and its metabolites were selected to thoroughly investigate their enantioselective toxicity and mechanisms at the molecular and cellular levels. Multispectral techniques revealed that the interaction between chiral PTC/PTCD and lysozyme resulted in the formation of a complex, leading to a change in the conformation of lysozyme. Meanwhile, the effect of different conformations of PTC/PTCD on the conformation of lysozyme differed, and its metabolites were able to exert a greater effect on lysozyme compared to prothioconazole. Moreover, the S-configuration of PTCD interacted most strongly with lysozyme. This conclusion was further verified by DFT calculations and molecular docking as well. Furthermore, the oxidative stress indicators within HepG2 cells were also affected by chiral prothioconazole and its metabolites. Specifically, S-PTCD induced more substantial perturbation of the normal oxidative stress processes in HepG2 cells, and the magnitude of the perturbation varied significantly among different configurations (P > 0.05). Overall, chiral prothioconazole and its metabolites exhibit enantioselective effects on lysozyme conformation and oxidative stress processes in HepG2 cells. This work provides a scientific basis for a more comprehensive risk assessment of the environmental behaviors and effects caused by chiral pesticides, as well as for the screening of highly efficient and less biotoxic enantiomeric monomers.

A highly selective and sensitive ICT-based Cu2+ fluorescent probe and its application in bioimaging.

Cu2+ is an essential trace element for the organism, but its excess can also cause irreversible damage to the organism. As such, a "Turn-Off" fluorescent probe DPAP for the specific detection of Cu2+ was successfully constructed. DPAP exhibits large Stokes shift (120 nm), fast reaction speed (1 min), low detection limit (15.2 nM), low toxicity, and good cell permeability. Cu2+ quenches the fluorescence of DPAP by blocking its intramolecular charge transfer process to achieve the detection of Cu2+ and has been confirmed by HRMS, 1H NMR and DFT calculations. Excitingly, the five-cycle detection of Cu2+ and the successful recovery of trace Cu2+ in environmental water samples fully demonstrate the potential of DPAP for practical applications. In particular, DPAP can observe the distribution and translocation patterns of exogenous Cu2+ in HeLa cells and zebrafish in real-time. This research concept has offered important theoretical support for the study of the environmental behavior of heavy metal ions.

Enantioselective effect of chiral prothioconazole on the conformation of bovine serum albumin.

As a typical chiral triazole fungicide, the enantioselective toxicity of prothioconazole to environmental organisms is of increasing concern. Herein, the binding mechanism of chiral PTCs to BSA was investigated by multi-spectral technique and molecular docking. Fluorescence titration and fluorescence lifetime experiments fully established that quenching BSA fluorescence by chiral PTCs is static quenching and could spontaneously bind to BSA. Hydrophobic interactions dominate the binding process of chiral PTCs to BSA. Differently, although both chiral PTCs and BSA have a primary binding site, the difference in chiral isomerism leads to a stronger binding ability of S-PTC than R-PTC. Both configurations of PTC can change the conformation of BSA and induce changes in the microenvironment around its amino acid residues, and the effect of S-PTC is more significant. Overall, S-PTC exhibited a more substantial effect on BSA structure relative to R-PTC. That is, S-PTC may lead to more potent potential toxicological effects on environmental organisms. This study provides a comprehensive assessment of the environmental behavior of chiral pesticides and their potential toxicity to environmental organisms at the molecular level and provides a theoretical basis for the screening of highly effective and biologically less toxic enantiomers of chiral pesticides.

Bioactivity, Uptake, and Distribution of Prothioconazole Loaded on Fluorescent Double-Hollow Shelled Mesoporous Silica in Soybean Plants.

Prothioconazole (PTC) has been widely utilized for plant fungal disease control, but its metabolite prothioconazole-desthio (PTC-d) exhibits reproductive toxicity. In the present study, carbon quantum dot (CQD)-modified fluorescent double-hollow shelled mesoporous silica nanoparticles (FL-MSNs) loaded with PTC, referred to as PTC@FL-MSNs, were constructed with an average size of 369 nm and a loading capacity of 28.1 wt %, which could increase the antifungal efficiency of PTC. In addition, upright fluorescence microscope and UPLC-MS/MS studies showed that PTC@FL-MSNs could be effectively transported via root uptake and foliar spray in soybean plants. Compared to a 30% PTC dispersible oil suspension agent, the PTC@FL-MSN treatment group showed higher concentrations (leaves: 0.50 > 0.48 mg/kg), longer half-lives for degradation (leaves: 3.62 > 3.21 d; roots: 3.39 > 2.82 d), and fewer metabolites. These findings suggest that sustained pesticide release and toxicity reduction are potential applications for PTC nanofungicide delivery technology.

A reversible CHEF-based NIR fluorescent probe for sensing Hg2+ and its multiple application in environmental media and biological systems.

Hg2+ poses a great threat to human health and the environment due to its bioaccumulation and permanent damage. Herein, a reversible CHEF-based near-infrared fluorescent probe 2-(3-((E)-4-((E)-4-(diethylamino)-2- hydroxybenzylidene)amino)styryl)-5,5-dimethylcyclohex-2-en-1-ylidene)propanedinitrile (DHEY) capable of specifically recognizing Hg2+ was constructed. DHEY exhibits advantages of large Stokes shift (157 nm), excellent selectivity, high sensitivity (LOD = 3.2 µg/L), and fast response efficiency (<3 min). Interestingly, DHEY can also realize rapid and effective detection of Hg2+ after being recycled 7 times. The successful recovery of trace Hg2+ in different environmental water samples fully demonstrates the potential of DHEY for actual applications. In particular, DHEY enables real-time observation of the distribution and translocation pattern of exogenous Hg2+ in HeLa cells and zebrafish. This work provides important theoretical support for investigating the fate of heavy metal ions in the environment using fluorescence techniques.

Construction of a novel fluorescent nanocarrier with double hollow shells for pH-controlled release of imidacloprid and its distribution and transport in bok choy.

Nanotechnology has been widely used in the field of pesticides. Integration of nano-pesticides and carbon dot fluorescence can fully utilize the potential for high admission of pesticides on leaves and convenience observation of its distribution and transport in the tissues. In the present study, a fluorescent mesoporous nanosilica with double hollow shells for loading imidacloprid (Im@FL-MSNs) was designed and synthesized. The physical and chemical properties of the imidacloprid nanocarriers were characterized by transmission electron microscopy (TEM), FT-IR spectroscopy, thermogravimetric analysis (TGA), X-ray photoelectron spectroscopy (XPS) and N2 adsorption/desorption. When the mass ratio of FL MSNs to imidacloprid is 6:5, Im@FL-MSNs exhibits good fluorescence properties, high loading efficiency (∼30%), great slow-release performance as well as pH controllability. Besides, Im@FL-MSNs can improve the ability of imidacloprid to adhere on the leaf surface of bok choy (Initial contact angled is greater than 80°ï¼‰. Importantly, Im@FL-MSNs did not reduce the biological activity of imidacloprid (LC50 (95% CI) = 1.43 mg/L). It was able to visually study the absorption and distribution of imidacloprid in bok choy plants, and provide theoretical and technical guidance for pesticide reduction.

Potential toxic effects of sulfonamides antibiotics: Molecular modeling, multiple-spectroscopy techniques and density functional theory calculations.

Sulfonamide antibiotics (SAs) are widely used in medicine, animal husbandry and aquaculture, and excessive intake of SAs may pose potential toxicity to organisms. The toxicological mechanisms of two classical SAs, sulfamerazine (SMR) and sulfamethoxazole (SMT), were investigated by molecular docking, DFT and multi-spectroscopic techniques using HSA and BSA as model proteins. The quenching of HSA/BSA endogenous fluorescence by SMR was higher than that by SMT due to the stronger binding effect of the pyrimidine ring on HSA/BSA compared to the oxazole ring, and that result was consistent with that predicted by DFT calculations. Thermodynamic parameters show that the binding of SAs to HSA/BSA is an exothermic process that proceeds spontaneously (ΔG < 0). Marker competition experiments illustrate that the binding site of SMR/SMT on serum albumin is located in subdomain IIIA. The combination of SAs and HSA/BSA is mainly realized by hydrogen bond and hydrophobic interaction, and the concept is also supported by molecular modeling. The reduced α-helix content of HSA/BSA induced by SMR/SMT indicates a greater stretching of the protein α-helix structure of the SMR/SMT-HSA/BSA. The results could provide useful toxicological information on the hazards of SAs in response to growing concern that SAs may pose a toxic threat to organisms.

Effects of plant morphology, vitamin C, and other co-present pesticides on the deposition, dissipation, and metabolism of chlorothalonil in pakchoi.

Pesticide residues have been a focus of attention of food safety. Different varietal pakchoi plants grown in open fields were studied to understand effects of morphology, leaf wax content, and vitamin C on the deposition, dissipation, and metabolism of chlorothalonil. The loose pakchoi plants and flat leaves were conducive to pesticide deposition, but not plants with erect leaves. Chlorothalonil on nine varieties of pakchoi dissipated in the first-order kinetic with T1/2 s of 1.4 ~ 2.0 days. Vitamin C in pakchoi could promote the dissipation of chlorothalonil. Carbendazim could significantly promote the dissipation of chlorothalonil on pakchoi. Interestingly, four metabolites of chlorothalonil were identified in the pakchoi and the metabolic pathway was predicted by DFT calculations. The risk assessment showed that pakchoi were safe for consumption after 10 days of application of the recommended dose. This work provides important information for the understanding of deposition, dissipation, and metabolism of chlorothalonil in pakchoi.

An ICT-Based Coumarin Fluorescent Probe for the Detection of Hydrazine and Its Application in Environmental Water Samples and Organisms.

As an inorganic small molecule pollutant, the toxicity and potential carcinogenicity of hydrazine (N2H4) are of increasing concern. In this work, A water-soluble fluorescent probe (OCYB) based on the intramolecular charge transfer (ICT) mechanism for the detection of hydrazine was designed and synthesized. Taking the advantage of 4-bromobutyryl as the recognition group, the high selectivity of OCYB to N2H4 was confirmed by steady-state fluorescence spectroscopy. The limit of detection (LOD) was calculated to be 78 nM in the DMSO-HEPES (pH 7.4) system. The detection mechanism was verified by NMR, HRMS and density functional theory (DFT) calculations. In addition, OCYB exhibits strong anti-interference ability and an "Off-On" fluorescence enhancement effect. Importantly, OCYB can be used to effectively monitor the fluorescence distribution of N2H4 in environmental water samples and organisms.

A novel ESIPT-based fluorescent probe with dual recognition sites for the detection of hydrazine in the environmental water samples and in-vivo bioimaging.

Hydrazine (N2H4), an important chemical intermediate, has been widely used in industrial production and agricultural life, but it has also caused environmental pollution. A novel ESIPT-based fluorescent probe with dual recognition sites, 2-(benzothiazole-2-yl)-1,4-imphenyl bis 4-bromobutyric acid (BRBA), was developed to selectively detect N2H4 under complex conditions. BRBA exhibits accurate detection for N2H4 with a good linear relationship ranging from 0 to 150 µM, and the LOD can reach 0.1 µM. Importantly, taking advantage of low cytotoxicity and a large Stokes shift, BRBA can be utilized to monitor environmental water samples and successfully applied to imaging HeLa cells and zebrafish.

Uptake, translocation and metabolism of imidacloprid loaded within fluorescent mesoporous silica nanoparticles in tomato (Solanum lycopersicum).

Fluorescence-labeling technology has been widely used for rapid detection of pesticides in agricultural production. However, there are few studies on the use of this technology to investigate pesticide uptake and transport in plants with fluorescent nanoparticle formulations. Here, we investigated uptake, transport, accumulation and metabolism of imidacloprid loaded in fluorescent mesoporous SiO2 nanoparticles (Im@FL-MSNs) in tomato plants, and compared the results with the pesticide application in granular formulation. The results revealed that Im@FL-MSNs applied via root uptake and foliar spray both could effectively transport in tomato plants and carry the imidacloprid to plant tissues. Neither Im@FL-MSNs nor imidacloprid was detected inside of tomato fruits from root uptake or foliar spray applications. Compared with the foliar application of granular formulation, imidacloprid in Im@FL-MSNs demonstrated a higher concentration in leaves (1.14 ± 0.07 mg/kg > 1.08 ± 0.04 mg/kg, 1.13 ± 0.09 mg/kg > 1.11 ± 0.02 mg/kg), longer half-life (0.84 d < 1.31 d, 0.90 d < 1.36 d) and small numbers of metabolites formed. These results suggest that mesoporous silica nanoparticles could serve as an effective and efficient pesticide carrier for achieving the high use efficiency in plant protection. The information is also helpful to guide the pesticide applications and assess the risks associated with environmental quality and dietary consumption of vegetables.

New insights into the interactions between humic acid and three neonicotinoid pesticides, with multiple spectroscopy technologies, two-dimensional correlation spectroscopy analysis and density functional theory.

The widespread use of neonicotinoid pesticides in agricultural production has caused pressure on the environment. In the present work, the interactions between humic acid (HA) and three neonicotinoid insecticides, dinotefuran, clothianidin and nitenpyram, were investigated by using multiple spectroscopy techniques combined with two-dimensional correlation spectroscopy analysis and density functional theory (DFT). Dinotefuran, clothianidin and nitenpyram could quench the endogenous fluorescence of HA through a static quenching process dominated by hydrogen bonds and van der Waals forces. According to the revised Stern-Volmer equation and DFT calculation, the binding abilities of the three pesticides with HA were ranked as dinotefuran < clothianidin < nitenpyram. The results of dynamic light scattering showed that neutral conditions were more conducive to the combination of HA and dinotefuran, clothianidin and nitenpyram. Through Fourier transform infrared spectroscopy (FTIR) combined with two-dimensional correlation analysis (2D-COS), the functional group with the strongest binding ability in the HA-dinotefuran, HA-clothianidin and HA-nitenpyram system was CH, CO and CO, respectively. The work will help to further understand the behavior of neonicotinoid pesticides in the environment.

A novel near-infrared fluorimetric method for point-of-care monitoring of Fe2+ and its application in bioimaging.

Iron is one of the essential trace elements in the human body, which is involved in many important physiological processes of life. The abnormal amount of iron in the body will bring many diseases. Therefore, a novel near-infrared fluorimetric method was developed. The method is based on a fluorescent probe (E)-4-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohex-1-en-1-yl)vinyl)-N, N-diethylaniline oxide (DDED) which uses N-oxide as a recognition group to real-time monitoring and imaging of Fe2+ in vivo and in vitro. The method exhibits excellent selectivity and high sensitivity (LOD = 27 nM) for Fe2+, fast reaction rate (< 4 min), extremely large Stokes shift (＞ 275 nm), low cytotoxicity. The strip test strongly illustrates the potential application of DDED in real environment. In particular, DDED has been successfully applied to real-time monitoring and imaging of Fe2+ in HepG2 cells and zebrafish. That is, the method has great potential for the detection of Fe2+ in living systems.

Hydrazine exposure: A near-infrared ICT-based fluorescent probe and its application in bioimaging and sewage analysis.

Hydrazine (N2H4) is an environment pollutant with high acute toxicity and potential carcinogenicity, and detection of N2H4 has attracted increasing attention. In the present study, a low toxicity near-infrared fluorescent probe (DCDB) based on the intramolecular charge transfer (ICT) principle was developed. The probe DCDB exhibits excellent selectivity and high sensitivity (LOD = 1.27 ppb) for N2H4, fast reaction rate (5 min), extremely large Stokes shift (160 nm). The color transformation of the DCDB-N2H4 system from purple to pink can be observed with the naked eye. The success of N2H4 test strips to detect trace N2H4 in actual sewage strongly illustrates the practical application potential of DCDB. Importantly, DCDB can be utilized to monitor the distribution of exogenous N2H4 in vivo and in vitro.

Interactions between Imidacloprid and Thiamethoxam and Dissolved Organic Matter Characterized by Two-Dimensional Correlation Spectroscopy Analysis, Molecular Modeling, and Density Functional Theory Calculations.

The heavy application of neonicotinoid insecticides in agricultural production has burdened the environment. In the present study, interactions of two neonicotinoid insecticides imidacloprid and thiamethoxam with dissolved organic matter (DOM) were investigated by spectroscopic techniques, molecular modeling, and density functional theory (DFT) calculations. The static mechanism of imidacloprid and thiamethoxam quenching the endogenous fluorescence of DOM was assessed through time-resolved analyses. During the binding process, a protein-like substance binds imidacloprid and thiamethoxam later than a humic-like substance, as analyzed by two-dimensional correlation spectroscopy, but more strongly than the humic-like substance, as suggested by molecular modeling and DFT calculations. The conformational changes of DOM are attributed to imidacloprid and thiamethoxam, as assessed with three-dimensional spectra. Fourier transform infrared spectroscopy indicated that DOM binds imidacloprid and thiamethoxam by hydroxyl, aliphatic C-H, amide I, and carboxyl to form stable DOM-imidacloprid and DOM-thiamethoxam complexes. Understanding the changes in the structural conformation of humic-like and protein-like substances with imidacloprid and thiamethoxam helps further understand the fate of the neonicotinoids in the environment.

An ICT-based fluorescent probe with a large Stokes shift for measuring hydrazine in biological and water samples.

As a strong reductant and highly active alkali, hydrazine (N2H4) has been widely used in chemical industry, pharmaceutical manufacturing and agricultural production. However, its high acute toxicity poses a threat to ecosystem and human health. In the present study, a ratiometric fluorescent probe for the detection of N2H4 was designed, utilizing dicyanoisophorone as the fluorescent group and 4-bromobutyryl moiety as the recognition site. 4-(2-(3-(dicyanomethylene)-5,5-dimethylcyclohex-1-enyl) phenyl 4-brobutanoate (DDPB) was readily synthesized and could specially sense N2H4 via an intramolecular charge transfer (ICT) pathway. The cyclization cleavage reaction of N2H4 with a 4-bromobutyryl group released phenolic hydroxyl group and reversed the ICT process between hydroxy group and fluorophore, turning on the fluorescence in the DDPB-N2H4 complexes. DDPB exhibits a low cytotoxicity, reasonable cell permeability, a large Stokes shift (186 nm) and a low detection limit (86.3 nM). The quantitative determination of environmental water systems and the visualization fluorescence of DDPB test strips provides a strong evidence for the applications of DDPB. In addition, DDPB is suitable for the fluorescence imaging of exogenous N2H4 in HeLa cells and zebrafish.

Multi-spectroscopic measurements, molecular modeling and density functional theory calculations for interactions of 2,7-dibromocarbazole and 3,6-dibromocarbazole with serum albumin.

2,7-Dibromocarbazole (2,7-DBCB) and 3,6-dibromocarbazole (3,6-DBCB) are emerging environmental pollutants, being potentially high risks to human health. In this study, interactions of the two compounds with human serum albumin (HSA) and bovine serum albumin (BSA) were investigated by molecular modeling, density functional theory calculations (DFT) and multispectral techniques. The static quenching interaction deduced in the fluorescence quenching experiment is confirmed by the time-resolved analyses. The interactions of the two compounds with HSA/BSA induce molecular microenvironment and conformation changes, as assessed by synchronous and 3D fluorescence spectra, together with a destruction of polypeptide carbonyl hydrogen bond network by circular dichroism and Fourier transform infrared analyses. The thermodynamic analysis indicated that the spontaneous interaction was hydrogen bonding and hydrophobic forces. The binding constant Ka at 298 K was 3.54 × 105 M-1 in HSA-2,7-DBCB, 6.63 × 105 M-1 in HSA-3,6-DBCB, 1.32 × 105 M-1 in BSA-2,7-DBCB and 2.17 × 105 M-1 in BSA-3,6-DBCB. These results indicates that 3,6-DBCB binds HSA/BSA more strongly than 2,7-DBCB, which was estimated with DFT calculations. Site marker competition experiments coupled with molecular modeling studies confirmed that both compounds bind HSA/BSA at site I (subdomain IIA). The results suggest that interactions between 2,7-DBCB and 3,6-DBCB with HSA and BSA may affect the normal physiological activities in human and animals.