Activatable fluorescent probes for imaging and diagnosis of rheumatoid arthritis

Rheumatoid arthritis(RA)is a systemic autoimmune disease that is primarily manifested as synovitis and polyarticular opacity and typically leads to serious joint damage and irreversible disability,thus adversely affecting locomotion ability and life quality.Consequently,good prognosis heavily relies on the early diagnosis and effective therapeutic monitoring of RA.Activatable fluorescent probes play vital roles in the detection and imaging of biomarkers for disease diagnosis and in vivo imaging.Herein,we review the fluorescent probes developed for the detection and imaging of RA biomarkers,namely reactive oxygen/nitrogen species(hypochlorous acid,peroxynitrite,hydroxyl radical,nitroxyl),pH,and cysteine,and address the related challenges and prospects to inspire the design of novel fluorescent probes and the improvement of their performance in RA studies.

Application and development of fluorescence probes in MINFLUX nanoscopy(invited paper)

The MINimal emission FLUXes(MINFLUX)technique in optical microscopy,widely recognized as the next innovative fluorescence microscopy method,claims a spatial resolution of 1-3 nm in both dead and living cells.To make use of the full resolution of the MINFLUX microscope,it is important to select appropriate fluorescence probes and labeling strategies,especially in living-cell imaging.This paper mainly focuses on recent applications and developments of fluorescence probes and the relevant labeling strategy for MINFLUX microscopy.Moreover,we discuss the deficiencies that need to be addressed in the future and a plan for the possible progression of MINFLUX to help investigators who have been involved in or are just starting in the field of super-resolution imaging microscopy with theoretical support.

Presheath formation and area design limit satellite-based Langmuir probes

In this article,the effect of the finite conductive surface area of a satellite on the use of satellite-based Langmuir probes is reviewed in light of the basic theory of asymmetric double Langmuir probes(ADLPs).Recent theoretical and experimental studies have discussed electron sheath/presheath formation and the electron Bohm criterion along with their implications for satellite-based Langmuir probes.The effects predicted by the latest theory of the electron Bohm criterion were not experimentally observed and the experimental results remain supportive of a critical area ratio(A_(L)/A_(S))_(crit)=(m_(i)/(2.3m_(e)))^(1/2)between the probe area A_(S)and the satellite area A_L as conventionally believed.A satellite-based Langmuir probe must satisfy this criterion to physically act as a single Langmuir probe.However,experimental investigations also found that high-energy electrons adversely affect(A_(L)/A_(S))_(crit)and a Langmuir probe's signal quality by giving additional electron current to A_(L).Based on these results,a number of limitations of the maximum probe area are derived when designing satellite-based Langmuir probes,with consideration of both the aim of the satellite and the plasma where the satellite-based probe works.These proposed measures are expected to only partially alleviate the effect of the inadequate satellite surface area on the application of satellite-based Langmuir probes.Using a larger satellite to carry a Langmuir probe remains the most viable means to obtain precise space plasma parameters.

Au@Ag Core-shell Nanorods Self-assembled on Polyelectrolyte Multilayers for Ultra-High Sensitivity SERS Fiber Probes

We demonstrated a chemical process in the fabrication of a SERS fiber probe with an ultrahigh sensitivity.The synthesis was carried out by preparing Au@Ag core-shell nanorods (Au@Ag-NRs) selfassembled on polyelectrolyte (PE) multilayers,for which Au@Ag-NRs were controlled by adjusting the silver layer thickness.The effect of silver layer thickness of Au@Ag-NRs on the SERS performance of the fiber probe was investigated.The SERS fiber probe shows the best performance when the silver layer thickness is controlled at 8.57 nm.Under the condition of optimizing silver layer thickness,the fiber probe exhibits ultra-high sensitivity (i e,10^(-10) M crystalline violet,CV),good reproducibility (i e,RSD of 3.5%) and stability.Besides,electromagnetic field distribution of the SERS fiber probe was also investigated.The strongest enhancement is found within the core of fiber,whereas a weakened electromagnetic field exists in the fiber cladding layer.The SERS fiber probe can be a good candidate in ultra-trace detection for biomedical and environmental areas.

Development of oligonucleotide probes for FISH karyotyping in Haynaldia villosa,a wild relative of common wheat

Haynaldia villosa is a wild relative of wheat and a valuable gene resource for wheat improvement.Owing to the limited number of probes available for fluorescence in situ hybridization(FISH),the resolution at which the karyotype of H.villosa can be characterized is poor,hampering accurate characterization of small segmental alien introgressions.We designed ten oligonucleotide probes using tandem repeats in DNA sequences derived from the short arm of H.villosa chromosome 6 V(6 VS).FISH with seven of them resulted in clear signals on H.villosa chromosomes.Using these,we constructed FISH karyotypes for H.villosa using oligo-6 VS-1 and oligo-6 VS-35 oligonucleotides and characterized the distribution of the two probes in five different H.villosa accessions.The new FISH probes can efficiently characterize H.villosa introgressions into wheat.

Hydroxylated graphene quantum dots as fluorescent probes for sensitive detection of metal ions

Highly sensitive methods are important for monitoring the concentration of metal ions in industrial wastewater.Here,we developed a new probe for the determination of metal ions by fluorescence quenching.The probe consists of hydroxylated graphene quantum dots(H-GQDs),prepared from GQDs by electrochemical method followed by surface hydroxylation.It is a non-reactive indicator with high sensitivity and detection limits of 0.01μM for Cu2+,0.005μM for Al3+,0.04μM for Fe3+,and 0.02μM for Cr3+.In addition,the low biotoxicity and excellent solubility of H-GQDs make them promising for application in wastewater metal ion detection.

An empirical model of the global distribution of plasmaspheric hiss based on Van Allen Probes EMFISIS measurements

Using wave measurements from the EMFISIS instrument onboard Van Allen Probes,we investigate statistically the spatial distributions of the intensity of plasmaspheric hiss waves.To reproduce these empirical results,we establish a fitting model that is a thirdorder polynomial function of L-shell,magnetic local time(MLT),magnetic latitude(MLAT),and AE*.Quantitative comparisons indicate that the model’s fitting functions can reflect favorably the major empirical features of the global distribution of hiss wave intensity,including substorm dependence and the MLT asymmetry.Our results therefore provide a useful analytic model that can be readily employed in future simulations of global radiation belt electron dynamics under the impact of plasmaspheric hiss waves in geospace.

Ultrasensitive Detection of Glyphosate Using CdTe Quantum Dots in Sol-Gel-Derived Silica Spheres Coated with Calix[6]arene as Fluorescent Probes

We have developed a simple method for the preparation of highly fluorescent and stable, water-soluble CdTe quantum dots in sol-gel-derived composite silica spheres which were coated with calix[6]arene. The resulting nanoparticles (NPs) were characterized in terms of UV, fluorescence and FT-IR spectroscopy and TEM. The results show that the new NPs display more intense fluorescence intensity and are more stable than its precursors of the type SiO2/CdTe. Under the optimum, the novel NPs exhibit a higher selectivity and ultrasensitive fluorescence probes for the determination of gly-phosate over other pesticides, the fluorescence intensity increase with the concentration of glyphosate in the range from 1.0 to 25.0 nmol/L and the detection limit is low to 0.0725 nmol/L. A mechanism is suggested to explain the inclusion process by a Langmuir binding isotherm.

Diagnosis of Hydrogen Plasma in a Miniature Penning Ion Source by Double Probes

Parameters of hydrogen plasma in a miniature Penning discharge ion source, including the electron temperature and the electron density, were measured by using double probes. The results indicate that the electron density increases and the electron temperature decreases with the increase in gas pressure and the discharge current. The electron temperature is about 5 - 9 eV and the electron density is 6.0× 10^13 ～ 1.2 × 10^14 m^-3 while the discharge current is in a range of 50 - 120 μA.

ACTIVATABLE SMART PROBES FOR MOLECULAR OPTICAL IMAGING AND THERAPY

Recent years have seen the design and implementation of many optical activatable smart probes.These probes are activatable because they change their optical properties and are smart because they can identify specific targets.This broad class of detection agents has allowed previously unperformed visualizations,facilitating the study of diverse biomolecules including enzymes,nucleic acids,ions and reactive oxygen species.Designed to be robust in an in vivo environment,these probes have been used in tissue culture cells and in live small animals.An emerging class of smart probes has been designed to harness the potency of singlet oxygen generating photosensitizers.Combining the discrimination of activatable agents with the toxicity of photosensitizers represents a new and powerful approach to disease treatment.This review highlights some applications of activatable smart probes with a focus on developments of the past decade.

Detection of HBV and HCV Coinfection by TEM with Au Nanoparticle Gene Probes

Gold（Au） nanoparticle HBV DNA or HCV cDNA gene probes were prepared and were used to detect HBV DNA and HCV RNA extracted from positive serum of patients with HBV and HCV coinfection directly by transmission electron microscopy （TEM）. PCR identifying HBV and HCV in serum of patients with HBV and HCV coinfection was established. Alkanethiol-modified oligonucleotide was bound with self-made Au nanoparticles to form nanoparticle HBV DNA or HCV cDNA gene probes through covalent binding of Au-S. HBV DNA and HCV RNA extracted from positive serum of patients with HBV and HCV coinfection was added to the detection system composed of nanoparticle HBV DNA and（or） HCV cDNA gene probes, The results showed that HBV DNA and HCV RNA could be specifically amplified by PCR. The zones of DNA amplification appeared in 431 bp and 323 bp respectively. When HBV DNA and HCV RNA extracted from positive serum of patients with HBV and HCV coinfection were added to the detection system, TEM displayed the nanoparticles self-assembled into large network aggregates. It was concluded that the detection of HBV and HCV coinfection by TEM was convenient and efficient with high specificity and sensitivity.

The Positions and Number of Flux Loops and Magnetic Probes for the HT-7U Superconducting Tokamak

The plasma shape and other paremeters such as /3P, li is important for the tokamak deveice where the plasma has a non-circular cross-section of sufficient elongation. The measuered signals of magnetic probes and flux loops are used to reconstruct the plasma shape and the current profile in device operation and plasma shape feed back control system. So the number and positions of magnetic probes and flux loops provides the basis of the plasma reconstruction. This paper instroduce how to use EFIT code (equilibrium fitting code) to determine the number and positions of the magnetic probes and flux loops. The simulation result is given also.

Microdosing cocktail study on the determination and pharmacokinetics of six hepatic cytochrome probes and their metabolites in rat

OBJECTIVE To describe a highly sensitive LC-ESI MSnmethod that was developed to simultaneously detect six CYP isoform-specific probes and their metabolites in rat plasma for microdosing cocktail study.METHODS After administration of a mixture of six probes(i.e.,a cocktail approach with caffeine 100μg·kg-1,tolbutamide100μg·kg-1,omeprazole 500μg·kg-1,dextromethorphan 500μg·kg-1,chlorzoxazone 50μg·kg-1and midazolam 100μg·kg-1)to SD rats.The plasma samples were extracted using ethyl acetate with diazepam and gliclazide as the IS.The assay was performed on an Agilent Eclipse Plus C18 column(2.1×50 mm,3.5μm).The mobile phase consisted of 0.01%formic acid(1 mmol·L-1ammonium formate)and acetonitrile.The flow rate was0.3 m L·min-1.The samples were analyzed by LC-20A&5500Qtrap ESI MSnin MRM mode.The MS/MS reaction selected 181.2/124.0 m/z ions for caffeine,195.2/138.2m/z for paraxanthine,269.1/170.0 m/z for tolbutamide,285.1/186.0 m/z for 4-hydroxytolbutamide,346.1/198.1m/z for omeprazole,362.2/214.2 m/z for 5-hydroxyomeprazole,272.3/147.1 m/z for dextromethorphan,258.2/157.0 m/z for dextrorphan,168.1/132.1 m/z for chlorzoxazone,326.1/291.2 m/z for midazolam,and 342.1/324.2m/z for 1′-hydroxymidazolam.RESULTS The datashowed that the method was with good linearity in the range of 0.2-200 ng·m L-1for caffeine,0.1-25 ng·m L-1for paraxanthine,0.05-100 ng·m L-1for omeprazole,0.01-25 ng·mL-1for 5-hydroxyomeprazole,0.1-200 ng·mL-1for dextromethorphan,0.05-12.5 ng·mL-1for dextrophan,0.2-200 ng·mL-1for midazolam,and 0.2-25 ng·mL-1for 1′-hydroxymidazolam,respectively.The stability%RSD for al probes was less than 15%and matrix effects in plasma on the ionization were negligible.CONCLUSION This highly sensitive and quantitative method allowed a pharmacokinetic study in subjects receiving doses 10-100 times lower than typical therapeutic doses.The established LCMS/MS method was suitable for pharmacokinetic study of this mixture cocktail probe group and could be applied deeply to CYP isoforms(1A2,2C9,2C19,2D6,2E1and 3A)research.

Synthesis of 3-hydroxyflavone fluorescent probes and study of their fluorescence properties

Direct measurement of dipole potential in biological membranes has been impossible and 3-hydroxyflavones(3HFs) have allowed detection of changes in dipole potential in biological systems.In the present study,sixteen derivatives of 3HF with aliphatic hydrocarbon chains of different lengths at 4′-position and 6-position were synthesized.The basic fluorescence properties of 3HFs are maintained in all the probes in terms of strong blue shift in maximum fluorescence emission wavelength and>100 fold increase in quantum yield in organic solvents and in dioleoylphosphatidylcholine(DOPC) small unilamellar vesicles(SUV) in comparison to in aqueous Hepes buffer(15 mmol/L,pH 7.4).More importantly,the ability of the new compounds to report dipole potential changes in biological systems are also maintained,since all the new probes showed spectrum properties that are similar to yet different from that of F4N1,which potentially may allow more sensitive measurement of the dipole potential change in membranes.

Cytotoxic T-cells as imaging probes for detecting glioma

Tumor vaccination using tumor-associated antigenprimed dendritic cells(DCs)is in clinical trials.Investigators are using patients’own immune systems to activate T-cells against recurrent or metastatic tumors.Following vaccination of DCs or attenuated tumor cells,clinical as well as radiological improvements have been noted due to migration and accumulation of cytotoxic T-cells(CTLs).CTLs mediated tumor cell killing resulted in extended survival in clinical trails and in preclinical models.Besides administration of primed DCs or attenuated or killed tumors cells to initiate the generation of CTLs,investigators have started making genetically altered T-cells(CTLs)to target specific tumors and showed in vivo migration and accumulation in the implanted or recurrent tumors using different imaging modalities.Our groups have also showed the utilization of both in vivo and in vitro techniques to make CTLs against glioma and used them as imaging probes to determine the sites of tumors.In this short review,the current status of vaccination therapy against glioma and utilization of CTLs as in vivo imaging probes to determine the sites of tumors and differentiate recurrent glioma from radiation necrosis will be discussed.

STRUCTURAL INVESTIGATION OF A NEW POLYORGANOSILOXANE CONTAINING NANOTUBULAR CAVITIES USING ORGANIC FLUORESCENT PROBES

Fluorescent molecules - p-dimethylaminobenzonitrile (DMABN) and trans-stilbene (TSB ) - were used as probes to investigate the inner structure of the nanotubular cavities contained in a new polyorganosiloxane (POS) which was prepared by the hydrosilylation coupling reaction of cis-isotactic ladderlike polyvinylsilsesquioxane (Vi-T) with 1,1,3,3-tetramethyldisiloxane (H-MM) as coupling agent in the presence of catalyst dicyclopentadienyldichloroplatinum (Cp2PtCl2). The results from FTIR spectra and fluorescence spectra in combination with molecular simulation reveal that the cross-section of the nanotubular cavity is nearly rectangular in shape, and is about 0.62 nm in width and about 0.38 nm in height. This work gains in-depth knowledge into the inner structure of the nanotubular cavities of POS and furthermore provides a guide to the selection of proper guest molecules in constructing functional supramolecular clathrate based upon POS.

PET probes beyond ^(18)F-FDG

During the past several decades,positron emission tomography（PET） has been one of the rapidly growing areas of medical imaging;particularly,its applications in routine oncological practice have been widely recognized.At present,^18F-fluorodeoxyglucose（^18F-FDG） is the most broadly used PET probe.However,^18F-FDG also suffers many limitations.Thus,scientists and clinicians are greatly interested in exploring and developing new PET imaging probes with high affinity and specificity.In this review,we briefly summarize the representative PET probes beyond ^18F-FDG that are available for patients imaging in three major clinical areas（oncology,neurology and cardiology）,and we also discuss the feasibility and trends in developing new PET probes for personalized medicine.

Ultrafast Terahertz Probes of Charge Transfer and Recombination Pathway of CH_3NH_3PbI_3 Perovskites

We use transient terahertz photoconductivity measurements to demonstrate that upon optical excitation of CH_3NH_3PbI_3 perovskite, the hole transfer from CH_3NH_3PbI_3 into the organic hole-transporting material（HTM）Spiro-OMe TAD occurs on a sub-picosecond timescale. Second-order recombination is the dominant decay pathway at higher photo-excitation fluences as observed in neat CH_3NH_3PbI_3 films. In contrast, under similar experimental conditions, second-order recombination weakly contributes the relatively slow recombination between the electrons in the perovskite and the injected holes in HTM, as a loss mechanism at the CH_3NH_3PbI_3/Spiro-OMe TAD interface. Our results offer insights into the intrinsic photophysics of CH_3NH_3PbI_3-based perovskites with direct implications for photovoltaic devices and optoelectronic applications.

Loading calcium fluorescent probes into protoplasts to detect calcium in the flesh tissue cells of Malus domestica

Cytosolic Ca^(2+)plays a key role in signal transduction in plants.Calcium imaging is the most common approach to studying dynamic changes in the cytoplasmic Ca^(2+)content.Here,we used mature‘Fuji’apples(Malus pumila Mill.)to obtain viable protoplasts from flesh tissue cells by enzymatic hydrolysis;then,three small-molecule fluorescent probes(fluo-8/AM,fluo-4/AM,and rhod-2/AM)were loaded into the protoplasts.All three Ca^(2+)fluorescent probes successfully entered the cytoplasm but did not enter the vacuole.Both the Ca^(2+)chelator(EGTA)and Ca^(2+)channel blocker(La3+)reduced the fluorescence intensity in the cytoplasm.The calcium ionophore A23187 increased the fluorescence intensity in the cytoplasm at 5μmol/L but decreased it at 50μmol/L.Additionally,A23187 reversed the fluorescence intensity in the cytoplasm,which was decreased by La3+.Ionomycin is also a calcium ionophore that can increase the fluorescence intensity of calcium in the cytoplasm.These results suggest that small-molecule Ca^(2+)fluorescent probes can be used to detect changes in cytosolic calcium levels in the cells of fruit flesh tissue.In addition,the optimum concentration of fluo-8/AM was determined to be 5μmol/L.This was the first time that protoplasts have been isolated from apple flesh tissue cells and small-molecule fluorescent probes have been used to detect calcium in the cytoplasm of flesh tissue cells.This study provides a new method to study calcium signal transduction in fruit flesh tissue.