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1.
J Chromatogr A ; 1736: 465411, 2024 Sep 30.
Article in English | MEDLINE | ID: mdl-39368194

ABSTRACT

Water, ubiquitous in analytical methods, is renowned for its fluorescence quenching properties, influencing techniques like fluorescence spectrophotometry or techniques with fluorescence detection. This study explores the impact of water (H2O) substitution for heavy water (D2O) on the fluorescence behavior of anthraquinones and anthracyclines. Anthraquinones and anthracyclines play crucial roles in pharmacy, serving as essential components in various therapeutic formulations, particularly in cancer treatment and other pharmacological interventions. Capillary electrophoresis (CE) with heavy water as the background electrolyte (BGE) solvent offers superior sensitivity to the separation and detection of these analytes. Experimental results demonstrate the improved detection limits and separation efficiency of selected anthraquinones rhein (RH), aloe-emodin (AE), and anthracyclines doxorubicin (DOX), epirubicin (EPI) and daunorubicine (DAU) in heavy water-based buffers, highlighting the potential of heavy water in advancing analytical chemistry.

2.
Nanomedicine (Lond) ; : 1-16, 2024 Oct 09.
Article in English | MEDLINE | ID: mdl-39382009

ABSTRACT

Aim: Fluorescence detection of breast and prostate cancer cells expressing Tn-antigen, a tumor marker, with Vicia villosa lectin (VVL)-labeled nanoparticles.Materials & methods: Breast and prostate cancer cells engineered to express high levels of Tn-antigen and non-engineered controls were incubated with VVL-labeled or unlabeled red dye-doped silica-coated polystyrene nanoparticles. The binding to cells was studied with flow cytometry, confocal microscopy, and electron microscopy.Results: Flow cytometry showed that the binding of VVL-labeled nanoparticles was significantly higher to Tn-antigen-expressing cancer cells than controls. Confocal microscopy demonstrated that particles bound to the cell surface. According to the correlative light and electron microscopy the particles bound mostly as aggregates.Conclusion: VVL-labeled nanoparticles could provide a new tool for the detection of Tn-antigen-expressing breast and prostate cancer cells.


[Box: see text].

3.
Methods Enzymol ; 705: 377-396, 2024.
Article in English | MEDLINE | ID: mdl-39389670

ABSTRACT

DNA and RNA can form various non-canonical secondary structures, including G-quadruplex (G4) and R-loops. These structures are considered transcriptional regulatory elements due to their enrichment at regulatory regions. During transcription, G-rich sequences in the non-template strand promote R-loop formation in the DNA template strand. These R-loops induce G4 structures in the non-template DNA strand, further stabilizing them. Additionally, the high rG: dC base-pairing within the R-loop contributes to the stability of DNA/RNA hybridization. Our previous study investigated the interplay between G4s and R-loops and its impact on transcription. We employed two techniques to demonstrate transcription-mediated G4 and R-loop formation. The single-molecule method allows us to detect intricate details of transcription initiation, elongation, and co-transcriptional R-loop and G4 formation. It provides a high-resolution view of the dynamic processes involved in transcriptional regulation. As an orthogonal approach, a gel-based assay enables the detection of the transcription-mediated R-loops and the RNA product. We can measure the progressive formation of R-loop and total RNA produced from transcription by analyzing gel electrophoresis patterns. In summary, these techniques provide valuable insights into the non-canonical nucleic acid structures and their impact on gene expression.


Subject(s)
DNA , G-Quadruplexes , R-Loop Structures , Transcription, Genetic , DNA/chemistry , DNA/genetics , Humans , RNA/chemistry , RNA/genetics , Single Molecule Imaging/methods
4.
Mikrochim Acta ; 191(11): 646, 2024 10 05.
Article in English | MEDLINE | ID: mdl-39367246

ABSTRACT

A reusable fluorescent nanoprobe was developed using DNA-functionalized metal-organic framework (MOF) for ratiometric detection of Hg2+. We utilized a zirconium-based MOF (UiO-66) to encapsulate tris(bipyridine) ruthenium(II) chloride (Ru(bpy)32+), resulting in Ru(bpy)32+@UiO-66 (RU) with red fluorescence. The unsaturated metal sites in UiO-66 facilitate the attachment of thymine-rich single-strand DNA (T-ssDNA) through Zr-O-P bond, producing T-ssDNA-functionalized RU complex (RUT). The T-ssDNA selectively binds to Hg2+, forming stable T-Hg2+-T base pairs and folding into double-stranded DNA, which permits the intercalation of SYBR Green I (SGI) and activates its green fluorescence. In the presence of Hg2+, SGI fluorescence increases in a dose-dependent manner, while Ru(bpy)32+ fluorescence remains constant. This fluorescence contrast enables RUT to serve as an effective ratiometric nanoprobe for Hg2+ detection, with a detection limit of 3.37 nM. Additionally, RUT demonstrates exceptional reusability due to the ability of cysteine to remove Hg2+, given its stronger affinity for thiol groups. The RUT was successfully applied to detect Hg2+ in real water samples. This work advances the development of ratiometric fluorescence nanoprobe based on DNA-functionalized MOFs.


Subject(s)
Fluorescent Dyes , Limit of Detection , Mercury , Metal-Organic Frameworks , Spectrometry, Fluorescence , Mercury/analysis , Mercury/chemistry , Fluorescent Dyes/chemistry , Metal-Organic Frameworks/chemistry , Spectrometry, Fluorescence/methods , Water Pollutants, Chemical/analysis , DNA/chemistry , Quinolines/chemistry , Benzothiazoles/chemistry , DNA, Single-Stranded/chemistry , Zirconium/chemistry , Diamines/chemistry , Phthalic Acids
5.
Article in English | MEDLINE | ID: mdl-39353101

ABSTRACT

The paramount importance of anticounterfeiting measures in safeguarding consumers from counterfeit products lies in their ability to ensure product safety and reliability. Advanced luminescent anticounterfeiting materials, particularly those responsive to multiple stimuli, afford a dynamic and multilayered security assurance. This study presents the synthesis of a novel material, Eu/Tb@GC-3, via postsynthetic modification, which exhibits notable photoluminescent properties with emission at 544 and 614 nm. The material demonstrates high selectivity and sensitivity in detecting Nitrofural and Enrofloxacin, with limits of detection at 0.0122 and 0.0280 µM, respectively. Furthermore, multistimulus responsive luminescent fibers and inks were developed, facilitating intelligent anticounterfeiting labels. The integration of these labels with back-propagation neural networks (BPNNs) significantly enhances pattern recognition and authentication capabilities, providing an efficacious strategy to combat counterfeit products and ensure consumer safety.

6.
J Fluoresc ; 2024 Oct 01.
Article in English | MEDLINE | ID: mdl-39352679

ABSTRACT

The rapid and accurate detection of programmed death-ligand 1 (PD-L1) expression is of great value in the diagnosis and treatment of tumors. ELISA-based traditional method is the gold standard for protein detection, but there are still some shortcomings, especially the antigen-antibody dependence, greatly increased the detection time and cost. This work constructed a label-free fluorescent probe for rapid and sensitive detection of PD-L1 using a truncated aptamer as recognition molecules and double-stranded DNA specific dyes (SYBR Green I) as signal units. After a series of optimization conditions, this probe has good detection capability for PD-L1 in buffer solution with the detection limit as low as 0.68 ng/mL. Due to the specific recognition ability of aptamer and target, this method also has good selectivity for PD-L1 detection. The recovery of PD-L1 in human serum samples ranges from 86.20 to 96.36%. Compared with other methods, this strategy does not need to be marked, and does not need other complex design and purification process, but simple operation process and strong anti-interference ability. The whole detection process can be completed within 20 min and has good application prospect. This work will provide reference for drug dosage and prognosis evaluation of specific tumor therapy.

7.
J Fluoresc ; 2024 Sep 20.
Article in English | MEDLINE | ID: mdl-39302493

ABSTRACT

Sulfadiazine (SD) is extensively utilized in agriculture, aquaculture, poultry, medical, and other industries. Its residues pose a threat to human health by entering the food chain and can also be released into the environment through animal feces and urine, leading to ecotoxicological pollution. Consequently, there is an urgent need to establish an efficient method for detecting SD residues in the environment. In this study, a novel two-probe fluorescence assay for determining SD in the environment, based on magnetic separation and real-time quantitative PCR-TaqMan probe technology, was successfully developed. In the experiment, SD served as the target substance, and an aptamer (Apt) with high affinity for SD was synthesized. Additionally, a non-fully complementary chain (Cdna) with favorable hybridization properties with the aptamer was designed and synthesized to create a magnetic probe of magnetic beads@Apt@Cdna. When SD was introduced, Apt specifically bound to SD with a hairpin structure and was released from the magnetic probe, allowing SD detection via the PCR-TaqMan method. Factors affecting the determination accuracy of this assay system, such as Apt concentration, SD standard solution pH, and incubation time, were optimized. Under the optimized conditions, the assay demonstrated high sensitivity for SD, with a detection limit of 2.34 × 10-5 ng/mL. Finally, the method was applied to detect SD in water samples from the Jialu River Basin in Zhengzhou City, yielding spiked recoveries of 88.82-117.05%. The results indicated that the detection system is a highly sensitive and specific method for determining SD residues in environmental water samples, showcasing its potential application in SD detection.

8.
Spectrochim Acta A Mol Biomol Spectrosc ; 325: 125177, 2024 Sep 19.
Article in English | MEDLINE | ID: mdl-39316855

ABSTRACT

BACKGROUND: MicroRNA (miRNA) has gradually become an emerging biomarker for early diagnosis and prognosis of various diseases due to its specific gene expression and high stability. With the development of molecular diagnosis and point-of-care testing (POCT) technology, developing simple, fast, sensitive, efficient, and low-cost miRNA sensors is of great significance for clinical applications and emergency rapid diagnosis. At present, entropy-driven toehold mediated chain displacement reaction, as a promising enzyme free isothermal amplification technique, is an important tool for ultra-sensitive biosensing applications. RESULTS: In this study, we used gold nanoparticles (AuNPs) as carriers and quenchers, modified them using self-assembled triple chain composite substrates AuNPs@A@B1/B2, and used dual reporter molecules for cascade cyclic amplification to amplify fluorescence signals, which proposed a fluorescent biosensor based on this reaction and build an intelligent fluorescence sensing platform for rapid detection of miRNA. We designed a highly specific self-programmable sensor using the acute ischemic stroke (AIS) biomarker miRNA-125a-5p as a sample, and achieved sensitive detection of miRNA in the range of 0.01 µM∼10 µ M under optimal conditions. It broke through the traditional detection limitations of weak signals and liberated the fluorescence detection environment. SIGNIFICANCE: In summary, this creative miRNA biosensor combined with POCT has demonstrated extraordinary detection potential, broad application prospects in the early diagnosis and prognosis monitoring of AIS, provides a novel miRNA universal detection strategy for the fields of biological and life sciences.

9.
J Fluoresc ; 2024 Sep 26.
Article in English | MEDLINE | ID: mdl-39325301

ABSTRACT

In recent years, perovskite quantum dots (PQDs) have successfully attracted widespread attention due to their excellent optical properties. However, the instability and toxicity problems of perovskite quantum dots are the main obstacles limiting their applications. In this work, bismuth-based perovskite quantum dots were synthesized by a ligand-assisted reprecipitation method, based on which a novel boric acid-functionalized bismuth-based non-toxic perovskite quantum dots fluorescent sensor (Cs3Bi2Br9-APBA) that can be stabilized in the ethanol phase was prepared by a boron affinity technique. Based on the covalent binding interaction of Cs3Bi2Br9-APBA with oxytetracycline (OTC), a highly selective and sensitive method for the detection of OTC was developed, which effectively solved the problems of poor stability and toxicity in the application of perovskite quantum dots. Under the optimal conditions, the fluorescence intensity of the synthesized Cs3Bi2Br9-APBA was linear with the concentration range of 0.1 ∼ 18 µM OTC, and the detection limit could reach 0.0802 µM. The fluorescence detection mechanism was explored and analyzed by spectral overlap analysis, suppression efficiency study of observed and corrected fluorescence, and fluorescence lifetime decay curve fitting, the mechanism of OTC detection by Cs3Bi2Br9-APBA was identified as the inner filter effect (IFE). In addition, the sensor successfully realized the quantitative detection of trace OTC in the environment, and our study provides a new idea for the preparation of green perovskite materials with high stability and selectivity.

10.
Phytochem Anal ; 2024 Sep 11.
Article in English | MEDLINE | ID: mdl-39261748

ABSTRACT

INTRODUCTION: Aflatoxins, potent carcinogens produced by Aspergillus species, present significant health risks and commonly contaminate herbal products such as Chrysanthemum morifolium. Detecting these toxins in C. morifolium proves challenging due to the complex nature of the herbal matrix and the fluctuating levels of toxins found in different samples. OBJECTIVES: This study aimed to develop and optimize a novel method for the detection of aflatoxins in C. morifolium using dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-fluorescence detection based on quality by design principles. METHODOLOGY: The method involved determining critical method attributes and parameters through the Plackett-Burman design, followed by optimization using the Box-Behnken design. Monte Carlo simulation was employed to establish a design space, which was experimentally verified. Method validation was performed to confirm accuracy, precision, and stability. RESULTS: The developed method exhibited excellent linearity (R2 > 0.9991) for aflatoxins B1, B2, G1, and G2 across a range of concentrations, with recovery rates between 85.52% and 102.01%. The validated method effectively quantified aflatoxins in C. morifolium under different storage conditions, highlighting the impact of temperature and storage time on aflatoxin production. CONCLUSION: This study successfully established a reliable and effective method for the detection of aflatoxins in C. morifolium, highlighting the importance of strict storage conditions to reduce aflatoxin contamination. Using a quality by design framework, the method demonstrated robustness and high analytical performance, making it suitable for routine quality control of herbal products.

11.
Talanta ; 281: 126892, 2024 Sep 16.
Article in English | MEDLINE | ID: mdl-39298805

ABSTRACT

Alkaline phosphatase (ALP) is abnormally expressed in some cancers and promotes the growth, metastasis, and invasion of cancer cells. The detection of ALP is of great significance for both pathological study and clinical detection. In this work, a europium (Eu)-based fluorescence detection sensor was prepared in a mild reaction condition. LaF3:Eu nanoparticles was mixed with ethylene imine polymer (PEI) and Ag+ ions. PEI was used as stabilizer and reducing agent, and Ag+ ions were reduced as molecular-like silver clusters (ML-Ag NCs). The fluorescence of LaF3:Eu nanoparticles was enhanced by ML-Ag NCs through energy transfer. When ascorbic acid 2-phosphate (AAP) was hydrolyzed to ascorbic acid (AA) in the presence of ALP, AA reduced Ag+ ions to silver nanoparticles (Ag NPs) and quenched the fluorescence of LaF3:Eu/PEI/Ag. The activity of ALP was detected by measuring the fluorescence intensity of Eu3+ at 618 nm. In the concentration range from 2.0 to 16.0 U/L, the fluorescence intensity ratio ((F0-F)/F0) had a linear relationship with the logarithm of ALP concentration. The limit of detection (LOD) was 1.3 U/L. Moreover, the ALP activity was detected successfully in cancer cells by this method. The sensing platform has application potential in the detection of ALP activity in biological systems.

12.
J Biomed Opt ; 29(9): 097001, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39224540

ABSTRACT

Significance: Although the depth detection limit of fluorescence objects in tissue has been studied, reports with a model including noise statistics for designing the optimum measurement configuration are missing. We demonstrate a variance analysis of the depth detection limit toward clinical applications such as noninvasively assessing the risk of aspiration. Aim: It is essential to analyze how the depth detection limit of the fluorescence object in a strong scattering medium depends on the measurement configuration to optimize the configuration. We aim to evaluate the depth detection limit from theoretical analysis and phantom experiments and discuss the source-detector distance that maximizes this limit. Approach: Experiments for detecting a fluorescent object in a biological tissue-mimicking phantom of ground beef with background emission were conducted using continuous wave fluorescence measurements with a point source-detector scheme. The results were analyzed using a model based on the photon diffusion equations. Then, variance analysis of the signal fluctuation was introduced. Results: The model explained the measured fluorescence intensities and their fluctuations well. The variance analysis showed that the depth detection limit in the presence of ambient light increased with the decrease in the source-detector distance, and the optimum distance was in the range of 10 to 15 mm. The depth detection limit was found to be ∼ 30 mm with this optimum distance for the phantom. Conclusions: The presented analysis provides a guide for the optimum design of the measurement configuration for detecting fluorescence objects in clinical applications.


Subject(s)
Phantoms, Imaging , Animals , Cattle , Limit of Detection , Spectrometry, Fluorescence/methods , Optical Imaging/methods
13.
Spectrochim Acta A Mol Biomol Spectrosc ; 325: 125111, 2024 Sep 08.
Article in English | MEDLINE | ID: mdl-39270366

ABSTRACT

In view of exploiting natural resources for designing of effectual materials in favor of detection and obliteration of water pollutants, a fluorescent nanomaterial (CDBHCF) based on biomass derived carbon dots (CDs) was constructed. The CDs and cobalt ferrite (CF) particles were anchored on boehmite (BH) which served as a support material for CDs. The CDBHCF nanocomposite was prepared via facile hydrothermal treatment for selective recognition of Methyl parathion (MP) pesticide and uranyl ions (UO22+). The corresponding structural, morphological and opto-electronic properties of the nanomaterials have been investigated by different physicochemical techniques. The fluorescent CDBHCF probe was employed to detect extremely low concentration of MP and UO22+ with detection limit of 22.4 nM and 4.4 nM, respectively. Ultimately, the proposed sensing platform was validated through real sample analysis. Besides, CDBHCF nanocomposite was utilized for photocatalytic abolition of Tetracycline (TC) in water samples. Initially, the impact of various operational parameters on the degradation efficiency, including catalyst dosage and initial pH were thoroughly examined. Under optimized conditions, the fabricated CDBHCF nanocomposite demonstrated excellent results for photocatalytic degradation of TC (92 % degradation in 120 min) under visible light illumination. Thus, the proposed strategy delivered an innovative insight for dual purpose of CDBHCF nanocomposite: as a fluorescent probe for real time monitoring and as a photocatalyst for removal of pollutants via simple photocatalytic degradation.

14.
aBIOTECH ; 5(3): 298-308, 2024 Sep.
Article in English | MEDLINE | ID: mdl-39279850

ABSTRACT

MicroRNAs (miRNAs) and short RNA fragments (18-25 nt) are crucial biomarkers in biological research and disease diagnostics. However, their accurate and rapid detection remains a challenge, largely due to their low abundance, short length, and sequence similarities. In this study, we report on a highly sensitive, one-step RNA O-circle amplification (ROA) assay for rapid and accurate miRNA detection. The ROA assay commences with the hybridization of a circular probe with the test RNA, followed by a linear rolling circle amplification (RCA) using dUTP. This amplification process is facilitated by U-nick reactions, which lead to an exponential amplification for readout. Under optimized conditions, assays can be completed within an hour, producing an amplification yield up to the microgram level, with a detection limit as low as 0.15 fmol (6 pM). Notably, the ROA assay requires only one step, and the results can be easily read visually, making it user-friendly. This ROA assay has proven effective in detecting various miRNAs and phage ssRNA. Overall, the ROA assay offers a user-friendly, rapid, and accurate solution for miRNA detection. Supplementary Information: The online version contains supplementary material available at 10.1007/s42994-024-00140-0.

15.
Spectrochim Acta A Mol Biomol Spectrosc ; 325: 125070, 2024 Sep 02.
Article in English | MEDLINE | ID: mdl-39232313

ABSTRACT

Four rhodamine 6G-PAH probes with pyrene (R6G-Pyr), anthracene (R6G-Ant), acenaphthene (R6G-Acp) or phenanthrene (R6G-PA) as fluorophore were designed and synthesized for Hg(II) detection. Probe R6G-PA, which had the lowest detection limit of 0.84 nmol/L, displayed the best fluorescence performance as compared to the other three probes. This type of probe had good anti-interference properties against most common metal ions except Cu(II). Metal Cu(II) had a certain quenching effect on the fluorescence generated by Hg(II), with a minimum detection limit of 0.31 nmol/L (for R6G-Acp), indicating its potential practicability for Cu(II) detection. The structure-fluorescence relationship was discussed based on density functional theory (DFT) calculations, and R6G-PA + Hg(II), which had the minimum dihedral angle between polycyclic aromatic rings and rhodamine spiro ring, produced the strongest π-π accumulation and provided the brightest fluorescence. Probe R6G-PA was successfully employed for fluorescence detection of Hg(II) in biological samples. Its solid-phase sensor PS@R6G-PA was developed by immobilizing R6G-PA on PS microspheres for the determination of Hg(II) in water and food samples, with excellent reproducibility and fluorescence "on/off" response. The relative error of the spiked recovery rate was less than 10 %.

16.
Carbohydr Polym ; 346: 122588, 2024 Dec 15.
Article in English | MEDLINE | ID: mdl-39245483

ABSTRACT

Aiming at dealing with organic and inorganic pollutants dissolved in aquatic environments, we introduce self-assembled fluorescent nanocomposite hydrogel based on a binary polysaccharide network (xanthan gum/chitosan) embedding nitrogen-doped carbon quantum dots not only as a hybrid solid optical sensor for detecting Cr(VI) ions but also to remove anionically charged contaminants Cr(VI) and methyl orange (MO) by acting as an adsorbent. This fluorescent nanocomposite achieved a detection limit of 0.29 µM when used to detect Cr(VI) and demonstrated a fluorescence quantum yield of 59.7 %. Several factors contributed to the effectiveness of the adsorption of Cr(VI) and MO in batch studies, including the solution pH, dosage of the adsorbent, temperature, initial contamination level, and contact time. Experimental results showed 456 mg/g maximum adsorption capacity at pH 4 for MO compared to 291 mg/g at pH 2 for Cr(VI) at 25 °C. In addition to conforming to Langmuir's model, Cr(VI) and MO's adsorption kinetics closely matched pseudo-second-order. Using thermodynamic parameters, the results indicate that Cr(VI) and MO adsorb spontaneously and exothermically. Recycling spent adsorbent for Cr(VI) and MO using NaOH at 0.1 M was possible; the respective adsorption efficiency remained at approximately 82.2 % and 83 % after the fifth regeneration cycle.

17.
Front Cell Infect Microbiol ; 14: 1454076, 2024.
Article in English | MEDLINE | ID: mdl-39233906

ABSTRACT

Introduction: Mycobacterium tuberculosis, the causative agent of human tuberculosis, poses a significant threat to global public health and imposes a considerable burden on the economy. However, existing laboratory diagnostic methods for M. tuberculosis are time-consuming and have limited sensitivity levels. Methods: The CRISPR/Cas system, commonly known as the "gene scissors", demonstrates remarkable specificity and efficient signal amplification capabilities. Enzymatic recombinase amplification (ERA) was utilized to rapidly amplify trace DNA fragments at a consistent temperature without relying on thermal cyclers. By integrating of CRISPR/Cas12a with ERA, we successfully developed an ERA-CRISPR/Cas12a detection system that enables rapid identification of M. tuberculosis. Results: The sensitivity of the ERA-CRISPR/Cas12a fluorescence and lateral flow systems was 9 copies/µL and 90 copies/µL, respectively. Simultaneously, the detection system exhibited no cross-reactivity with various of respiratory pathogens and non-tuberculosis mycobacteria, demonstrating a specificity of 100%. The positive concordance rate between the ERA-CRISPR/Cas12a fluorescence system and commercial qPCR was 100% in 60 clinical samples. Meanwhile, the lateral flow system showed a positive concordance rate of 93.8% when compared to commercial qPCR. Both methods demonstrated a negative concordance rate of 100%, and the test results can be obtained in 50 min at the earliest. Discussion: The ERA-CRISPR/Cas12a system offers a rapid, sensitive, and specific method that presents a novel approach to laboratory diagnosis of M. tuberculosis.


Subject(s)
CRISPR-Cas Systems , Mycobacterium tuberculosis , Nucleic Acid Amplification Techniques , Sensitivity and Specificity , Mycobacterium tuberculosis/genetics , Mycobacterium tuberculosis/isolation & purification , Humans , Nucleic Acid Amplification Techniques/methods , Tuberculosis/diagnosis , Tuberculosis/microbiology , Recombinases/metabolism , Recombinases/genetics , Molecular Diagnostic Techniques/methods , Bacterial Proteins/genetics , DNA, Bacterial/genetics , CRISPR-Associated Proteins/genetics , Endodeoxyribonucleases
18.
Talanta ; 281: 126850, 2024 Sep 12.
Article in English | MEDLINE | ID: mdl-39276573

ABSTRACT

MicroRNAs (miRNAs) are currently recognized as important biomarkers for the early diagnosis and prognostic treatment of cancer. Herein, we developed a simple and label-free method for the multiplex detection of miRNAs, based on entropy-driven circuit (EDC) amplification and non-gel sieving capillary electrophoresis-LED induced fluorescence detection (NGCE-LEDIF) platform. In this system, three different lengths of fuel chains were designed to catalyze three EDC, targeting miRNA-21, miRNA-155, and miRNA-10b, respectively. In the presence of target miRNA, the EDC cycle amplification reaction was triggered, generating numerous stable double-strands products (F-DNA/L-DNA). Since the three miRNAs correspond to three different lengths of F-DNA/L-DNA, they can be easily isolated and detected by NGCE. This strategy has good sensitivity, with detection limits of 68 amol, 292.2 amol, and 394 amol for miRNA-21, miRNA-155, and miRNA-10b, respectively. Additionally, this method has good specificity and can effectively distinguish single-base mismatches of miRNA. The recoveries of the three miRNAs in deproteinized healthy human serum ranged from 91.28 % to 108.4 %, with a relative standard deviation (RSD) of less than 7.9 %. This method was further applied to detect cellular miRNAs in human breast cancer (MCF-7) cell extracts, revealing an up-regulation of miRNA-21, miRNA-155, and miRNA-10b in MCF-7 cells. The successful spiked recovery in human serum and RNA extraction from MCF-7 cells underscores the practicality of this method. Therefore, this strategy has broad application prospects in biomedical research.

19.
Food Chem ; 460(Pt 3): 140754, 2024 Dec 01.
Article in English | MEDLINE | ID: mdl-39121762

ABSTRACT

The determination of biogenic amines (BAs) in alcoholic beverages is crucial for assessing their health impact, ensuring beverage quality, and guaranteeing safety. Herein, a rapid one-pot derivatization/magnetic solid-phase extraction (OPD/MSPE) method was proposed using 6-aminoquinolinyl-N-hydroxysuccinimide carbamate as the derivatization reagent and magnetic hydroxyl-functionalized multi-walled carbon nanotubes as the extraction material. Integration of derivatization and extraction steps simplifies the sample preparation process, taking only three minutes and eliminating the need for centrifugation by utilizing magnetic sorbent. The resulting desorption solution was directly analyzed by high-performance liquid chromatography-fluorescence detection (HPLC-FLD) without any evaporation or reconstitution steps. The integrated OPD/MSPE-HPLC-FLD method demonstrates excellent linearity (R2 > 0.992), accuracy (relative recoveries: 85.1-109.2%), precision (RSDs≤9.7%) and detection limits (limits of detection: 0.3-2 ng/mL). It has been successfully applied to determine free BAs in various alcoholic beverages, including red wine, Baijiu, Huangjiu, and beer. This method enables rapid, sensitive and precise analysis of BAs in alcoholic beverages.


Subject(s)
Alcoholic Beverages , Biogenic Amines , Solid Phase Extraction , Solid Phase Extraction/methods , Chromatography, High Pressure Liquid , Alcoholic Beverages/analysis , Biogenic Amines/analysis , Biogenic Amines/isolation & purification , Limit of Detection , Fluorescence
20.
Int J Biol Macromol ; 278(Pt 3): 134883, 2024 Oct.
Article in English | MEDLINE | ID: mdl-39168203

ABSTRACT

Uranium is a key element in the nuclear industry, whose accidental release causes health and environmental problems. In this paper, a protein-directed fluorescent sensor with aggregation-induced emission characteristics (gold nanoclusters@ovalbumin, AuNCs@OVA) was synthesized for the detection of UO22+ with high sensitivity and selectivity. The sensor exhibited good fluorescence stability, and its fluorescence intensity could be selectively enhanced by UO22+. Based on FT-IR and XPS analyses, the increase in fluorescence intensity of AuNCs@OVA after the addition of UO22+ was attributed to aggregation induced by the complexation between UO22+ and the amino, carboxyl, hydroxyl, and phosphate groups of ovalbumin. The detection limit was determined to be 34.4 nM, and the sensor showed excellent ion selectivity for UO22+. In combination with a smartphone program, the sensor could realize the real-time detection of UO22+ in a quantitative and portable way.


Subject(s)
Gold , Metal Nanoparticles , Uranium , Gold/chemistry , Metal Nanoparticles/chemistry , Uranium/analysis , Limit of Detection , Spectrometry, Fluorescence/methods , Ovalbumin/chemistry , Ovalbumin/analysis , Fluorescent Dyes/chemistry , Ions/analysis , Uranium Compounds/analysis
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