Theoretical Investigation of a Coumarin Fluorescent Probe for Distinguishing the Detection of Small-Molecule Biothiols.

Monitoring the level of biothiols in organisms would be beneficial for health inspections. Recently, 3-(2'-nitro vinyl)-4-phenylselenyl coumarin as a fluorescent probe for distinguishing the detection of the small-molecule biothiols cysteine/homocysteine (Cys/Hcy) and glutathione (GSH) was developed. By introducing 4-phenyselenium as the active site, the probe CouSeNO2/CouSNO2 was capable of detecting Cys/Hcy and GSH in dual fluorescence channels. Theoretical insights into the fluorescence sensing mechanism of the probe were provided in this work. The details of the electron excitation process in the probe and sensing products under optical excitation and the fluorescent character were analyzed using the quantum mechanical method. All these theoretical results would provide insight and pave the way for the molecular design of fluorescent probes for the detection of biothiols.

Theoretical investigation on a simple turn on fluorescent probe for detection of biothiols based on coumarin unit.

The discovery of a simple and efficient detection method for biothiols would be scientifically significant due to the crucial role of them in various physiological processes. Recently, a simple fluorescent probe, DEMCA-NBSC, based on coumarin fragments, was developed by Ding et al., and provided an efficient way for real-time sensing of biothiols both in vivo and vitro. Theoretical insights to the fluorescence sensing mechanism of the probe were provided in this work. Details of the electron transfer process in the probe under optical excitation and the fluorescent character of the probe were analyzed using a quantum mechanical method. All these theoretical results could inspire the development of a highly convenient and efficient fluorescent probe to sense biothiols both in vivo and vitro.

A Single Organic Fluorescent Probe for the Discrimination of Dual Spontaneous ROS in Living Organisms: Theoretical Approach.

Single-organic-molecule fluorescent probes with double-lock or even multi-lock response modes have attracted the attention of a wide range of researchers. The number of corresponding reports has rapidly increased in recent years. The effective application of the multi-lock response mode single-molecule fluorescent probe has improved the comprehensive understanding of the related targets' functions or influences in pathologic processes. Building a highly efficient functional single-molecule fluorescent probe would benefit the diagnosis and treatment of corresponding diseases. Here, we conducted a theoretical analysis of the synthesizing and sensing mechanism of this kind of functional single-molecule fluorescent probe, thereby guiding the design and building of new efficient probes. In this work, we discuss in detail the electronic structure, electron excitation, and fluorescent character of a recently developed single-molecule fluorescent probe, which could achieve the discrimination and profiling of spontaneous reactive oxygen species (ROS, •OH, and HClO) simultaneously. The theoretical results provide insights that will help develop new tools for fluorescent diagnosis in biological and medical fields.

A Novel Fluorescence Probe Based on Azamonardine for Detecting and Imaging Cysteine in Cells and Zebrafish with High Selectivity and Sensitivity.

A novel fluorescent probe based on azamonardine (Aza) fluorophore was designed and synthesized for the highly selective detection of cysteine (Cys) in vivo and in vitro. After reacting with acryloyl chloride, the fluorescence of Aza is effectively quenched, resulting in the formation of the Aza-acryl probe. Upon the addition of Cys, the ester bond of Aza-acryl is cleaved, releasing a new compound (Compound 1) with strong fluorescence, thereby achieving fluorescence turn-on detection of Cys. The structure of Aza-acryl was characterized using X-ray crystallography and NMR spectroscopy. Additionally, density functional theory was employed to elucidate the quenching mechanism of the acyl group on the Aza. Aza-acryl exhibits high selectivity towards Cys and distinguishes it from other biothiols such as homocysteine (Hcy) and glutathione (GSH). The mechanism of Aza-acryl for detecting Cys was investigated through HPLC, NMR spectroscopy, high-resolution mass spectrometry, and reaction kinetics experiments. Aza-acryl demonstrates excellent imaging capabilities for Cys in cells and zebrafish, providing a reliable and selectable tool for the detection and imaging of Cys in biological systems.

Theoretical Insights into a Near-Infrared Fluorescent Probe NI-VIS Based on the Organic Molecule for Monitoring Intracellular Viscosity.

So many biological functional disorders and diseases, such as atherosclerosis, hypertension, diabetes, Alzheimer's disease, as well as cell malignancy are closely related with the intracellular viscosity. A safe and effective intracellular viscosity detecting method is desired by the biomedical community. Recently, a novel near-infrared fluorescent probe NI-VIS with a twisting intramolecular charge transfer mechanism was developed. The capability of this probe to visualize the viscosity variation in cirrhotic liver tissues and map the micro viscosity in vivo were testified using an experiment. In this work, the twisting intramolecular charge transfer mechanism and fluorescent properties of the probe NI-VIS were studied in detail under quantum mechanical method. The low energy barrier among the different conformations of the probe indicated the occurrence of twisting intramolecular charge transfer due to the rotation of the aryl group in the probe molecule while within the low viscosity environment. The electronic structure analysis on different probe conformations revealed the electron transfer process of the probe under optical excitation. All these theoretical results could provide insights into understand in greater depth the principles and build highly effective fluorescent probe to monitor the viscosity in biological samples.

Theoretical investigation on FRET strategy of ratio metric fluorescent probe sensing hydrogen sulfide.

The level of hydrogen sulfide (H2S) in human body is related to many diseases, such as Alzheimer's disease, Down syndrome, etc. Therefore, the detection of H2S level in biological systems is very important and has attracted great attention from scientific and clinical researchers. Understanding the design and working mechanism of fluorescent probes for H2S level detection is important for building new highly efficient fluorescent probe. The mechanisms of a recently reported efficient small molecule fluorescent probe based on the Fluorescence Resonance Energy Transfer (FRET) were investigated thoroughly in this work. The theoretical results would provide the insights for designing new efficient and multi-functional fluorescent probe applicable in the biological systems.

Electronic structure and interaction in CH4@C60: a first-principle investigation.

CH4@C60 was the first example within which an organic molecule has been embedded in C60. CH4 can rotate freely in the molecular cage, and the carbon skeleton structure of the C60 has no obvious deformation. The electronic structure of CH4@C60 and interaction between C60 and CH4 were studied under quantum mechanical calculation method. The different reaction sites on C-C bonds in C60 and the weak Van der Waals interaction between CH4 and C60 were shown clearly. These results and the orbital interaction between CH4 and C60 were helpful for understanding and further application of this unique biggest organic molecule CH4 contained in C60 structure so far.

Automatic and sensitive detection of West Nile virus non-structural protein 1 with a portable SERS-LFIA detector.

A portable surface-enhanced Raman scattering (SERS)-lateral flow immunoassay (LFIA) detector has been developed for the automatic and highly sensitive detection of West Nile virus (WNV) non-structural protein 1 (NS1) and actual WNV samples. Au@Ag nanoparticles (Au@Ag NPs) labeled with double-layer Raman molecules were used as SERS tags to prepare WNV-specific SERS-LFIA strips. On this platform, the WNV-specific antigen NS1 protein was quantitatively and sensitively detected. The detection limit for the WNV NS1 protein was 0.1 ng/mL, which was 100-fold more sensitive than visual signals. The detection limit for inactivated WNV virions was 0.2 × 102 copies/µL. The sensitivity of the SERS-LFIA detector was comparable to that of the fluorescence quantitative reverse transcription-polymerase chain reaction assay. The prepared SERS-LFIA strips exhibited high sensitivity and good specificity for WNV. Thus, the strips developed herein have clinical application value. Moreover, the portable SERS-LFIA detector enabled automatic and rapid detection of the SERS-LFIA strips. The platform established herein is expected to make a substantial contribution to the diagnosis and control of outbreaks of emerging infectious diseases, including WNV.

Duplex-specific nuclease signal amplification-based fluorescent lateral flow assay for the point-of-care detection of microRNAs.

MiRNAs play important regulatory roles in numerous biological processes and serve as significant biomarkers for the development and prognosis of several diseases. Their unique characteristics, such as short size, high sequence homology among family members, low abundance, and easy degradability, have hindered their specific and highly sensitive detection. Herein, a duplex-specific nuclease (DSN)-assisted target recycling signal amplification-based fluorescent lateral flow assay was demonstrated for the point-of-care detection of cancer-related miRNA-21. In this assay, digoxin/biotin-labeled DNA probes were selectively cleaved by the DSN enzyme in the rounds of hybridization with the miRNA-21 target and cleavage cycle. Subsequently, the resulting mixture, containing the miRNA-21 target and intact and cleaved DNA probes, was loaded onto the lateral flow strip with digoxin antibody-conjugated quantum dot nanobeads and the streptavidin-coated test line. The increase in the proportion of cleaved DNA probes can induce a weakened response signal, which is directly associated with the amount of the miRNA target. Thus, highly sensitive quantification of miRNA-21 was achieved at a low limit of detection of 0.16 pM within 2 h of assay time. Assay specificity toward miRNA-21 was validated by testing several other miRNAs, including let-7b, let-7d, miRNA-141, and miRNA-200a. Moreover, the assay can quantify miRNA-21 spiked in human serum samples with acceptable recovery values, thus indicating its considerable clinical feasibility.

Portable and multiplexed lateral flow immunoassay reader based on SERS for highly sensitive point-of-care testing.

A portable surface-enhanced Raman scattering (SERS)-based lateral flow immunoassay (LFIA) reader with multiplexed detection was developed using an integrated LFIA reaction column. The proposed LFIA reader was designed to simultaneously detect multiple samples or samples with multiple biomarkers. With the integrated LFIA reaction column, we achieved the specific detection of alpha-fetoprotein (AFP), carcinoembryonic antigen (CEA), and prostate-specific antigen (PSA) with a detection limit of 0.01 ng/mL, which was three orders of magnitude lower than that of the visual signal. We also investigated the uniformity of channels based on an eight-channel integrated LFIA reaction column. The relative standard deviation values of the SERS intensity of the eight-channel for measuring the AFP, CEA, and PSA antigens at 1323 cm-1 were 13%, 4.8%, and 5%, respectively. We detected 45 clinical serum samples of the three antigens using the proposed portable SERS-based LFIA reader to further confirm its applicability to clinical samples. The SERS signals of the positive sera were higher than those of the negative sera and their thrice standard deviation. This result indicated the practicality of the developed integrated reaction column and the proposed portable and multiplexed Raman reader. This work provides a new high-sensitivity, multiplexed, and automated SERS-based LFIA detector for use in the point-of-care setting.

Rapid Enrichment and Ultrasensitive Detection of Influenza A Virus in Human Specimen using Magnetic Quantum Dot Nanobeads Based Test Strips.

Influenza A virus (IAV) possesses a high infectivity and pathogenicity, and can lead to severe respiratory infection with similar symptoms caused by some other common respiratory viruses. Lateral flow assay (LFA) has been widely deployed in remote settings as a rapid and reliable approach for point-of-care detection of infectious pathogens. However, it still remains challenging to detect IAV virions using LFA from clinical samples such as nasopharyngeal or throat swabs, because their various components and high viscosity can decrease flow velocity and lead to the nonspecific adsorption of nanoparticle labels on the sensing membrane. Herein, we demonstrated a magnetic quantum dot nanobeads (MQBs) based LFA for magnetic enrichment and fluorescent detection of IAV virions in clinical specimens. In this study, MQBs were synthesized and then conjugated with IAV-specific antibody to efficiently enrich IAV virions from complex biological matrix, but also serve as highly bright fluorescent probes in lateral flow strips. This assay can achieve quantitative detection of IAV virions with a low limit of detection down to 22 pfu mL-1 within 35 minutes, and show good specificity between influenza B virus and two adenovirus strains. Furthermore, the presented platform was able to directly detect IAV virions spiked in nasopharyngeal swab dilution, indicating its stability and feasibility in clinical applications. Thus, this point-of-care detection platform holds great promise as a broadly applicable approach for the rapid diagnosis of influenza A.

Probing the glass transition in reversible cross-linked polymer composites.

Understanding the nature of glass transition is still a great challenge. Glass transition is widely observed in many glassy materials; however, it has never been unambiguously observed in reversible cross-linked polymer, which is an ideal model of the percolation process. Herein, we report the synthesis of a reversible cross-linked polymer incorporated with four-armed Diels-Alder (DA) dynamic covalent bonds, and the robust experimental observation of percolation-induced glass transition in this reversible four-armed cross-linked polymer (DAMF1). Temperature-modulated differential scanning calorimetry (TMDSC) experiment results clearly revealed the presence of a glass transition along with an endothermic or exothermic peak associated with DA/retro-DA (RDA) reaction related to the reconstitution/disassociation of the DAMF1's four-armed cross-linked network. In situ 13C variable-temperature solid-state NMR experiments further confirmed the DA/RDA reaction during glass transition at a molecular level. The above experimental results provide a direct experimental evidence for the recently developed percolation model of glass transition, which provides new insights into the nature of glass transition.

Development of ratiometric sensing and white light-harvesting materials based on all-copper nanoclusters.

Herein, we developed a special strategy for the fast sensitization of red emitting copper nanoclusters with the assistance of green emitting copper nanoclusters. Compared to most previous methods based on AIE, which do not maintain the water solubility or tiny size of nanoclusters, the charming features of the copper nanoclusters were retained after the fabrication. Furthermore, the product was employed for the detection of sulphide, which revealed its ratiometric sensing ability in water since the ratio of the intensity change for green and red emission was related to the sulphide concentration. In addition, after the addition of Zn2+, the green and red emission was either enhanced or quenched via the corresponding mechanism. This enables the facile fabrication of promising white light-harvesting materials since the species of the emitting color can be simply tuned.

Study on the Glass Transition Process of Polymer System Using Differential Scanning Calorimetry and Fourier Transform Infrared Spectroscopy.

The change in the infrared spectrum of polymer samples with temperature and their differential scanning calorimetry (DSC) experimental results are analyzed. According to the van't Hoff equation at constant pressure, the changes in the absorbance ratio corresponding to high and low vibrational states are calculated, and the apparent enthalpy differences of the vibration energy states transformation of the characteristic group can be obtained. From the experimental results, we can find that characteristic vibration modes of a chemical group in a polymer are under the influence of the glass transition process of the polymer with a different extent. The characteristic vibration modes of the same chemical group behave differently due to the influence of the polymer system at which the chemical moiety is situated.

A Single Molecular Diels-Alder Crosslinker for Achieving Recyclable Cross-Linked Polymers.

A triol-functional crosslinker combining the thermoreversible properties of Diels-Alder (DA) adducts in one molecule is designed, synthesized, and used as an ideal substitute of a traditional crosslinker to prepare thermal recyclable cross-linked polyurethanes with excellent mechanical properties and recyclability in a very simple and efficient way. The recycle property of these materials achieved by the DA/retro-DA reaction at a suitable temperature is verified by differential scanning calorimetry and in situ variable temperature solid-state NMR experiments during the cyclic heating and cooling processes. The thermal recyclability and remending ability of the bulk polyurethanes is demonstrated by three polymer processing methods, including hot-press molding, injection molding, and solution casting. It is notable that all the recycled cross-linked polymers display nearly invariable elongation/stress at break compared to the as-synthesized samples. Further end-group functionalization of this single molecular DA crosslinker provides the potential in preparing a wide range of recyclable cross-linked polymers.

[Application of a new design of cryo-jaw and its biomechanical evaluation in rat achilles tendon in vitro].

This study was aimed to design a new, accurate and easy-to-use water bath cryo-jaw, and try to solve the problems met in small animals achilles tendon mechanical testing. The muscle-tendon-bony units were fixed in the clamps. SD rats achilles tendon were randomly divided into group A and B. Group A was tested by the newly designed water bath cryo-jaw, while group B was treated by non-water bath cryo-jaw. The mechanical tests revealed that non of the samples of the newly-designed water bath cryo-jaw in group A slipped and fell off, and the achilles tendons were in a physiologically active state, but one of the group B samples slipped and fell off, and the others had the frozen phenomenon obviously. The maximum stress, fracture displacement and Young's modulus of the rats in group A were significantly different compared to those in group B (P < 0.05). In conclusion, the new water bath cryo-jaw has more advantages than traditional ones. It exhibits a good simulation in vivo in the environmental conditions for testing the mechanical properties of the achilles tendon.