A biomimetic approach to conjugate vitamin B6 cofactor with the lysozyme cocooned fluorescent AuNCs and its application in turn-on sensing of zinc(II) in environmental and biological samples.

This communication focusses on the synthesis of red fluorescent lysozyme cocooned gold nanoclusters (Lyso-AuNCs) that have been successfully applied for the selective and specific recognition of the vitamin B6 cofactor pyridoxal-5'-phosphate (PLP). The red fluorescence of Lyso-AuNCs showed remarkable color change to yellow upon conjugation with PLP due to the formation of a Schiff base between the free -NH2 present in the lysozyme and the -CHO group of PLP. The developed PLP conjugated Lyso-AuNCs (PLP_Lyso-AuNCs) was applied for the selective turn-on recognition of Zn2+ ions in aqueous medium. The yellow fluorescence of PLP_Lyso-AuNCs exhibited significant enhancement at 475 nm in the presence of Zn2+ producing bluish-green fluorescence attributed to the complexation-induced aggregation of nanoclusters. The nanoprobe exhibits nanomolar limit of detection for Zn2+ ions (39.2 nM) and the practicality of the nanoprobe was validated in various environmental water samples and biological plasma, urine, and beetroot extract, with fairly good recovery percent. Also, the system was successfully implemented for the intracellular detection and monitoring of Zn2+ in live HeLa cells. Graphical abstract Applications of red emitting lysozyme cocooned gold nanoclusters (Lyso-AuNCs) for the selective recognition of the vitamin B6 cofactor pyridoxal-5'-phosphate (PLP) and the conjugated nano-assembly PLP_Lyso-AuNCs for turn-on detection of Zn2+ ions in various environmental and biological samples.

Functional DNA-Zn2+ coordination nanospheres for sensitive imaging of 8-oxyguanine DNA glycosylase activity in living cells.

Sensitive monitoring of human 8-oxyguanine DNA glycosylase (hOGG1) activity in living cells is helpful to understand its function in damage repair and evaluate its role in disease diagnosis. Herein, a functional DNA-Zn2+ coordination nanospheres was proposed for sensitive imaging of hOGG1 in living cells. The nanospheres were constructed through the coordination-driven self-assembly of the entropy driven reaction (EDR) -deoxyribozyme (DNAzyme) system with Zn2+, where DNAzyme was designed to split structure and assembled into the EDR system. When the nanospheres entered the cell, the competitive coordination between phosphate in the cell and Zn2+ leaded to the disintegration of the nanospheres, releasing DNA and some Zn2+. The released Zn2+ acted as a cofactor of DNAzyme. In the presence of hOGG1, the EDR was completed, accompanied by fluorescence recovery and the generation of a complete DNAzyme. With the assistance of Zn2+, DNAzyme continuously cleaved substrates to produce plenty of fluorescence signals, thus achieving sensitive imaging of hOGG1 activity. The nanospheres successfully achieved sensitive imaging of hOGG1 in human cervical cancer cells (HeLa), human non-small cell lung cancer cells and human normal colonic epithelial cells, and assayed changes in hOGG1 activity in HeLa cells. This nanospheres may provide a new tool for intracellular hOGG1 imaging and related biomedical studies.

The Dynamics of Lamin a During the Cell Cycle.

Lamin proteins play an essential role in maintaining the nuclear organization and integrity; and lamin A, in particular, plays a major role in the whole volume of the nuclear interior. Although the nucleus is highly organized, it is rather dynamic, it affects crucial nuclear processes and its organization must change as cells progress through the cell cycle. Although many aspects of these changes are already known, the role of lamin A during nuclear assembly and disassembly as well as its underlying mechanisms remains controversial. Here we used live cells imaging and Continuous Photobleaching (CP) method to shed light on the dynamics and mechanisms of lamin A during the cell cycle, combined with imaging flow cytometry measurements, which provides the high-throughput capabilities of flow cytometry with single-cell imaging. As a major analysis tool, we used spatial correlation algorithm for allocating the distribution of lamin A, chromatin and tubulin, as well as their mutual colocalization. Furthermore, we analyzed the distribution of lamin A along the nuclear lamina and in the nucleus interior during the cell cycle. Our results indicate that at the beginning of the cell division that include prophase, metaphase and anaphase, lamin A is distributed throughout the cytoplasm and its concentration in the chromosomal regions is reduced, whereas the spatial correlation between lamin A and tubulin is increased. It implies that lamin A also disassembled in the whole cellular volume. At the telophase and early G1, lamin A is concentrated in the whole volume of the newly formed nuclei of the daughter cells and it assembles to the lamina. We also explored the functional aspects of lamin A during the cell cycle and its binding to the chromatin versus the freely diffusion form. We found that the fraction of the bound proteins of lamin A in the S phase increased, relative to the G1 phase, which means that during replication, the concentration of lamin A on the chromatin increases. All these results shed light on the function of lamin A throughout the cell cycle.

Development of a method for the detection of Cu2+ in the environment and live cells using a synthesized spider web-like fluorescent probe.

A macrocyclic Schiff base fluorescent probe [1,2-phenylenediamine-2,6-pyridinedialdehyde macrocyclic Schiff base] (BP-MSB) based on 2,6-pyridinedialdehyde was synthesized for use in the detection of Cu2+ in environmental water samples and live cells imaging by the method of specific recognition. The free fluorescent probe BP-MSB shows strong fluorescence in DMSO/H2O. The probe shows high sensitivity and selectivity for Cu2+ through "turn-off" fluorescence response in DMSO/H2O buffer solution (pH = 6.5), with a detection limit of 0.83 nM, which is far below the maximum allowable drinking water content of 20.0 µM specified by the US Environmental Protection Agency. The BP-MSB fluorescence quenching method was used for the determination of Cu2+ in Xiang Jiang water samples and tap-water. Furthermore, addition of the same number of moles of ethylene diamine tetraacetic acid (EDTA) can realize the reversible recognition of Cu2+ by the probe BP-MSB. Most importantly, the fluorescence imaging of live cells after incubation of BP-MSB with GM12878 cells showed good imaging performance, confirming the sensitivity of the fluorescent probe BP-MSB in vivo. The probe was also used to form an analog logic gate. This probe has the advantages of good stability, simple operation and high selectivity, which provides a broad prospect for environmental monitoring, intracellular detection and practical application of POCT.

Synthesis and Discovery of Schiff Base Bearing Furopyrimidinone for Selective Recognition of Zn2+ and its Applications in Cell Imaging and Detection of Cu2.

A simplefuro [2,3-d]pyrimidinone-based Schiff base FPS was synthesized via aza-Wittig reaction and structure elucidation was carried out by spectroscopic studies FT-IR, 1H NMR, and 13C NMR and mass spectrometry. FPS showed weak fluorescence emission in methanol and the selectivity of FPS to different metal ions (Mn2+, Ca2+, Fe2+, Fe3+, Mg2+, Al3+, Ba2+, Ag+, Co2+, Na+, K+, Cu2+, Zn2+, Pb2+, Bi3+) were studied by absorption and fluorescence titration. The results show that FPS has selective fluorescence sensing behavior for Zn2+ ions and the limit of detection (LOD) was calculated to be 1.19 × 10-8 mol/L. Moreover, FPS-Zn2+ acts as a metal based highly selective and sensitive new chemosensor for Cu2+ ions and the LOD was calculated to be 2.25 × 10-7 mol/L. In accordance with the results and theoretical calculations, we suspected that the binding mechanisms of FPS to Zn2+ and Cu2+ were assigned to be the cooperative interaction of Zn2+(Cu2+)-N.

5-Hydroxymethylfurfural modified rhodamine B dual-function derivative: Highly sensitive and selective optical detection of pH and Cu(2+).

A dual-function optical chemosensor (RBF) was designed and easily synthesized by condensation reaction of 5-Hydroxymethylfurfural and rhodamine B hydrazide. RBF exhibited highly sensitive, highly selective and quick response to acidic pH. The fluorescence intensity of RBF exhibited a more than 41-fold increase within the pH range from 7.50 to 3.73 with a pKa value of 5.02, which could be successfully applied to monitor intracellular pH in living PC12 cells and HeLa cells. Additionally, the spectroscopy of UV-Vis and EDTA-adding experiments indicated that RBF was a highly selective and reversible colorimetric chemosensor for Cu(2+) in Tris-HCl (10mM, pH=7.2) aqueous buffer solution as well as other metal ions had no obvious interference. Moreover, RBF has been successfully applied to detect Cu(2+) in real water samples.

A high performance Schiff-base fluorescent probe for monitoring Au(3+) in zebrafish based on BODIPY.

We designed and synthesized a mono-Schiff-base fluorescent probe (Probe 1) based on a boron-dipyrromethene (BODIPY) dye. By investigating the recognition of Au(3+) through an irreversible C=N bond hydrolysis reaction, Probe 1 exhibited higher properties such as acting as a "naked eye" probe, stability to pH, fast-response of 90s, a lower detection limit of 60 nM, stronger antijamming capability, and better live-cells imaging with low cytotoxicity compared with other probes. Even in relatively high temperatures, Probe 1 maintained its own excellent characteristic. More importantly, this is the first time that one chemosensor could be successfully applied to Au(3+) imaging in zebrafish, which demonstrated the performance that Probe 1 exhibited wonderful organism permeability.

A novel fluorescent "turn-on" chemosensor for nanomolar detection of Fe(III) from aqueous solution and its application in living cells imaging.

An electronically active and spectral sensitive fluorescent "turn-on" chemosensor (BTP-1) based on the benzo-thiazolo-pyrimidine unit was designed and synthesized for the highly selective and sensitive detection of Fe(3+) from aqueous medium. With Fe(3+), the sensor BTP-1 showed a remarkable fluorescence enhancement at 554 nm (λex = 314 nm) due to the inhibition of photo-induced electron transfer. The sensor formed a host-guest complex in 1:1 stoichiometry with the detection limit down to 0.74 nM. Further, the sensor was successfully utilized for the qualitative and quantitative intracellular detection of Fe(3+) in two liver cell lines i.e., HepG2 cells (human hepatocellular liver carcinoma cell line) and HL-7701 cells (human normal liver cell line) by a confocal imaging technique.