Boronate affinity-based template-immobilization surface imprinted quantum dots as fluorescent nanosensors for selective and sensitive detection of myricetin

February 21, 2022 0 Comments

  • In order to prepare a kind of efficient fluorescence sensors for determination of cis-diol-containing flavonoids, novel imprinted quantum dots for myricetin (Myr) were prepared based on boronate affinity-based template-immobilization surface imprinting. The obtained boronate affinity-based surface imprinted silica (imprinted APBA-functionalized CdTe QDs) was used as recognition elements. The quantum dots were used as signal-transduction materials. Under the optimum conditions, according to fluorescence quenching of imprinted APBA-functionalized CdTe QDs by Myr, the imprinting factor (IF) for Myr was evaluated to be 7.88. The result indicated that the boronate affinity functionalized quantum dots coated with imprinted silica were successfully prepared.
  • The prepared imprinted APBA-functionalized CdTe QDs exhibited good sensitivity and selectivity for Myr. The fluorescence intensity was inversely proportional to the concentration of Myr in the 0.30-40 μM concentration range. And its detection limit was obtained to be 0.08 μM. Using the fluorescence sensors, the detection of Myr in real samples was successfully carried out, and the concentration of Myr in green tea and apple juice samples was evaluated to be 2.26 mg/g and 0.73 mg/g, respectively.
  • The recoveries for the spiked green tea and apple juice samples were 95.2-105.0% and 91.5-111.0%, respectively. This study also provides an efficient fluorescent detection method for cis-diol-containing flavonoids in real samples.

Application of quantitative fluorescent polymerase chain reaction analysis for the rapid confirmation of trisomy 13 of maternal origin in a pregnancy with fetal holoprosencephaly, cyclopia, polydactyly, omphalocele and cell culture failure

Objective: We present the application of quantitative fluorescent polymerase chain reaction (QF-PCR) for the rapid confirmation of trisomy 13 of maternal origin in a pregnancy with fetal holoprosencephaly (HPE), cyclopia, polydactyly, omphalocele and cell culture failure.
Case report: A 21-year-old, gravida 2, para 0, woman was referred for termination of the pregnancy at 17 weeks of gestation because of the abnormal ultrasound finding of alobar HPE. The pregnancy was subsequently terminated, and a 118-g malformed male fetus was delivered with cyclopia, bilateral postaxial polydactyly of the hands and ruptured omphalocele. Postmortem cell culture of the placental tissue and umbilical cord was not successful. The parental karyotypes were normal. QF-PCR analysis using the polymorphic DNA markers of D13S1810, D13S790 and D13S251 on the DNA extracted from placenta, umbilical cord and parental bloods showed trisomy 13 of maternal origin.
Conclusion: Perinatal diagnosis of concomitant HPE, polydactyly and omphalocele should raise a suspicion of fetal trisomy 13. QF-PCR analysis is useful for rapid confirmation of trisomy 13 and the parental origin especially under the circumstance of cell culture failure, and the information acquired is very useful for genetic counseling of the parents.

A new HClO-activated “turn-off” mitochondria-targetable NIR fluorescent probe for imaging of osteoarthritis in vivo

Hypochlorous acid (HClO) as a biomarker of inflammation has been implicated in redox signaling and combating microbial infection. Therefore, it is of great significance to develop an efficient method for detection and analysis of HClO in osteoarthritis. Herein, a new”turn-off” mitochondria-targetable NIR fluorescent probe, NIR-ClO, was reported for specific analysis and imaging of osteoarthritis response-related HOCl levels in vitro and in vivo.
In the presence of HClO, due to the specific HOCl-triggered C = C bond cleavage reaction, NIR-ClO obtained a high sensitive and selective fluorescence “On-Off” response toward HClO with a good limit of detection(LOD) as low as 28.3 nM, and showed a fast response time (<60 s) , which allow it to be used for detection of HClO under a simulated physiological condition. In addition, NIR-ClO was successfully used to imaging of HClO in living RAW264.7 cells and osteoarthritis model rat. The results suggest that NIR-ClO is a robust tool for future studies on diagnosis osteoarthritis.

Preparation of layering-structured magnetic fluorescent liposomes and labeling of HepG2 cells

Background: At present, surgical resection and chemotherapy are still the main treatments for hepatocellular carcinoma and other cancers, but the curative effect and survival rate are not ideal.
Objective: In this study, we aim to prepare a carrier with low toxicity, high biocompatibility and targeted transport for the treatment of hepatocellular carcinoma.
Methods: CdSe quantum dots (QDs) modified with oleic acid were synthesized. Then hydrophobic CdSe QDs and hydrophilic super-paramagnetic Fe3O4 particles were encapsulated into different layers of liposomes to form magnetic fluorescent liposomes (MFLs). MFLs in the aqueous would quickly drift towards the external magnet and the entire process was clearly observed with fluorescence microscope. The fluorescence spectra revealed that the fluorescence properties of MFLs were similar to that of CdSe QDs.
Results: QDs had an average size of 3.32 nm with good fluorescence properties. The size of MFLs was about 100 nm (transmission electron microscopy (TEM) analysis showed the average size of MFLs was about 82.8 nm and dynamic light scattering (DLS) detection showed 111.9 nm). After being cultured with MFLs for 8 h, HepG2 cells were labeled by MFLs and good fluorescence images were obtained. MTT analysis also expressed their good biocompatibility.
Conclusion: The prepared MFLs had multi-function and could be used as ideal drug carriers.

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All-in-one: A Robust Fluorescent Fusion Protein Vector Toolbox for Protein Localization and BiFC Analyses in Plants

  • Fluorescent tagging protein localization (FTPL) and bimolecular fluorescence complementation (BiFC) are popular tools for in vivo analyses of the subcellular localizations of proteins and protein-protein interactions in plant cells. The efficiency of fluorescent fusion protein (FFP) expression analyses is typically impaired when the FFP genes are co-transformed on separate plasmids compared to when all are cloned and transformed in a single vector. Functional genomics applications using FFPs such as a gene family studies also often require the generation of multiple plasmids. Here, to address these needs, we developed an efficient, modular all-in-one (Aio) FFP (AioFFP) vector toolbox, including a set of fluorescently labeled organelle markers, FTPL and BiFC plasmids, and associated binary vectors.
  • This toolbox uses Gibson assembly (GA) and incorporates multiple unique nucleotide sequences (UNSs) to facilitate efficient gene cloning. In brief, this system enables convenient cloning of a target gene into various FFP vectors or the insertion of two or more target genes into the same FFP vector in a single-tube GA reaction. This system also enables integration of organelle marker genes or fluorescently fused target gene expression units into a single transient expression plasmid or binary vector.
  • We validated the AioFFP system by testing genes encoding proteins known to be functional in FTPL and BiFC assays. In addition, we performed a high-throughput assessment of the accurate subcellular localizations of an uncharacterized rice CBSX protein subfamily. This modular UNS-guided GA-mediated AioFFP vector toolkit is cost-effective, easy to use and will promote functional genomics research in plants.

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