A novel rhodamine-based fluorescent probe for high selectively determining cysteine in lysosomes

The three primary biothiols are cysteine (Cys), glutathione (GSH), and homocysteine (Hcy) which plays key functions in a variety of biological processes. Intracellular and extracellular microenvironments are perfectly regulated in the living system, while in case of any fluctuations in the redox, buffer level may lead to several diseases. The unbalance in the extracellular compartment is influenced by the changes in intracellular metabolism and these are correlated with cellular processes such as death, differentiation, and proliferation, in the live system. Therefore, a study was performed to develop copper-based fluorescence probes for the sensitive and selective detection of extracellular and intracellular targets such as cysteine (Cys) and glutathione (GSH) respectively. Conversely, the quenched fluorescence of complex (Probes + Cu) by the coordination of either Cys or GSH, leads to retained Cu2+ ion due to its approximate binding with Cys or GSH. In this review, we have segmented the reported probes according to the fluorophore used, analyzed their photophysical characteristics, and highlighted the advantages and disadvantages of each (limit of detection, mechanism). 

Targeted cysteine and glutathione detection in extra/intracellular systems by copper-based fluorescent imaging probes

Acinetobacter is a vast bacterial genus comprising of numerous species with variable characteristics. The enigma associated with clinical strains which have been implicated in many nosocomial outbreaks has prompted the need for continuous research on pathogens like Acinetobacter baumannii and members of the ACB complex. However, numerous species of Acinetobacter genus possess diverse metabolic capabilities and have the potential for a plethora of industrial and environment-based applications. Therefore, a comprehensive review on the entire genus including many under-represented topics would contribute extensive information to the scientific community indulged in Acinetobacter research. The current review is a unique compilation which attempts to provide the latest update on the genus covering its clinical as well as ecological aspects. Moreover, it is the first study of its kind which focuses on the entire genus and elaborates on the phylogenetic relationships, pathogenesis and virulence mechanisms followed by emerging biotechnological applications with future directions.

The multifaceted genus Acinetobacter: from infection to bioremediation.

Although paraquat (PQ) is one of the most extensively used herbicides worldwide, it is hazardous to the environment and the public due to its extreme toxicity towards non-target organisms especially animals and humans. Therefore, developing a detection system for selectively and sensitively detecting PQ remains a great challenge. This report demonstrates the design of a novel highly fluorescent rhodamine-based probe fabricated by rationally inserting adamantane-modified rhodamine into a host cucurbit[7]uril (CB[7]) with a host-guest interaction induced spirocyclic ring-opening process. This host-guest binary complex (1&CB[7]) had a stronger fluorescence intensity than rhodamine B, which was exploited for PQ detection. Notably, complex 1&CB[7] displayed high sensitivity toward PQ, with an excellent nanomolar limit of detection of 10 nM. It was determined that this unique detection mechanism involved a process of self-assembly, disassembly, and reassembly, thereby providing a reversible fluorescence response to PQ. With such merits, we successfully quantified and mapped PQ in actual water samples, live cells, and various agricultural products. We anticipated that the constructed complex 1&CB[7] could acts as a potential alternative for PQ tracking in various applications, highlighting a new paradigm for engineering host-guest probes with spirocyclic ring-opening process for analyte detection. 

Engineering a novel rhodamine-based fluorescent probe using host-guest interactions for reversible, selective, and sensitive detection of herbicide paraquat

In this study, a novel rhodamine-based optically and electrochemically active chemosensor, integrated with a p-DMAC moiety, demonstrated extremely selective identification of Au3+ ions relative to other metal species, including (Li+, Na+, K+, Ba2+, Ca2+, Mg2+, Co2+, Mn2+, Zn2+, Pb2+, Ni2+, Fe2+, Hg2+, Fe3+, Cd2+, Pd2+, Al3+, Cr3+, Cu2+, and nitrate salt of Ag+). These compounds demonstrated a novel and outstanding aggregation-induced emission enhancement (AIEE) behavior by aggregating in DMF/H2O medium. Furthermore, the degree of quenching was varying linearly with a Au3+ concentration from 0 to 40 nM, with a lower detection limit by RH-DMAC nanoaggregates of 118.79 picomolar (40.35 ppm). The Stern-Volmer plots, Job's plot, Benesi-Hildebrand plot, 1H NMR titrations, ESI-mass, and FTIR all revealed significant interactions between the sensor and Au3+. Moreover, the proposed electrochemical sensor afforded a linear correlation before the peak current and concentration of Au3+ in the range of 0-40 nM, with a detection limit of 483.73 pM or 164.36 ppt (by cyclic voltammetry method) and 298.0 pM or 101.24 ppt (by the Differential Pulse Voltammetry method). Furthermore, the proposed sensing assay was used to measure Au3+ ion in spiked water samples (tap, drinking, waste, and river water), achieving acceptable accuracy and precision with high recovery rates. Furthermore, RH-DMAC-coated fluorescence paper test strips were designed for on-site Au3+ detection. Apart from this, the use of smartphone-based RGB (Red Green Blue) color analysis shortened the operating process, accelerated the detection technique, and provided a novel methodology for the instantaneous, real-time examination of Au3+ in real water samples.

AIEE Active Azomethine-Based Rhodamine Derivative For Ultrasensitive Multichannel Detection of Au3+ Through a Fluorimetrically, Electrochemically, and RGB-Based Sensing Assay.

A Rhodol-Derived Probe for Intracellular Biothiols Imaging and Rapid Labelling of Sulfhydryl-Containing Proteins

A rapid and convenient strategy to monitor the productivity of biomanufacturing is essential for the research in optimizing relevant bioprocesses. In this work, we have developed a fluorescein-derived probe (FL-DT) that reacts rapidly with thiol groups via 1,4-Michael addition reaction of the sulfhydryl to unsaturated ketone and releases fluorescence. FL-DT specifically forms fluorescent adduct with two adjacent thiols in a protein of interest (POI), making the probe a reliable tool for protein quantification. The production of xylanase fused with a short di-Cys tag was then successfully monitored and quantified with FL-DT in Escherichia coli system under different protein expression conditions, providing useful information for optimizing the bioprocess. Our work provides a convenient and efficient strategy for POI labeling and monitoring bioproduction. 

Fluorescent Labeling of
 Dicysteine‐Tagged
 Peptide for Monitoring and Optimization of Protein Bio‐production in Bacteria

Objectives
By combining the metabolomics and computational biology, to explore the relationship between metabolic phenotype and pathological stage in esophageal cancer patients, to find the mechanism of metabolic network disturbance and develop a new method for fast preoperative clinical staging.


Methods
A prospective cohort study (from April 2013 to January 2016) was conducted. The preoperative patients from Sichuan Provincial People's Hospital, who were diagnosed with esophageal cancer from May 2013 to April 2014 were included, and their serum samples were collected to detect 1H-nuclear magnetic resonance (NMR) metabolomics for the purpose of drawing the metabolic fingerprinting in different stages of patients with esophageal cancer. The data were processed with these methods-principal components analysis: partial least squares regression and support vector machine, for the exploration of the enzyme-gene network regulatory mechanism in abnormal esophageal cancer metabolic network regulation and to build the quantitative prediction model of esophageal cancer staging in the end. All data were processed on high-performance computing platforms Matalab. The comparison of data had used Wilcoxon test, variance analysis, χ2 test and Fisher exact test.


Results
Twenty patients with different stages of esophageal cancer were included; and their serum metabolic fingerprinting could differentiate different tumor stages. There were no difference among the five teams in the age (F=1.086, P>0.05), the body mass index (F=1.035, P>0.05), the distance from the incisors to tumor (F=1.078, P>0.05). Among the patients with different TNM stages, there was a significant difference in plasma metabolome. Compared to ⅡB, ⅢA, Ⅳstage patients, increased levels of butanone, ethanol amine, homocysteine, hydroxy acids and estriol, together with decreased levels of glycoprotein, creatine, choline, isobutyricacid, alanine, leucine, valine, were observed inⅠB, ⅡA stage patients. Four metabolic markers (ethanol amine, hydroxy-propionic acid, homocysteine and estriol) were eventually selected. gene ontology analysis showed that 54 enzymes and genes regulated the 4 key metabolic markers. The quantitative prediction model of esophageal cancer staging based on esophageal cancer NMR spectrum were established. Cross-validation results showed that the predicted effect was good (root mean square error=5.3, R2=0.47, P=0.036).


Conclusions
The systems biology approaches based on metabolomics and enzyme-gene regulatory network analysis can be used to quantify the metabolic network disturbance of patients with advanced esophageal cancer, and to predict preoperative clinical staging of esophageal cancer patients by plasma NMR metabolomics.


Key words: 
Esophageal neoplasms; Metabolomics; Surgery; Computational Biology; Metabolic networks and pathways; Gene Ontology; Neoplasm staging

[Using (1)H-nuclear magnetic resonance metabolomics and gene ontology to establish pathological staging model for esophageal cancer patients].

The pollution caused by improper handling of plastics has become a global challenge. In addition to recycling plastics and using biodegradable plastics, an alternative solution is to seek efficient methods for degrading plastics. Among them, the methods of using biodegradable enzymes or microorganisms to treat plastics have attracted increasing attention because of its advantages of mild conditions and no secondary environmental pollution. Developing highly efficient depolymerizing microorganisms/enzymes is the core for plastics biodegradation. However, the current analysis and detection methods cannot meet the requirements for screening efficient plastics biodegraders. It is thus of great significance to develop rapid and accurate analysis methods for screening biodegraders and evaluating biodegradation efficiency. This review summarizes the recent application of various commonly used analytical techniques in plastics biodegradation, including high performance liquid chromatography, infrared spectroscopy, gel permeation chromatography, and determination of zone of clearance, with fluorescence analysis techniques highlighted. This review may facilitate standardizing the characterization and analysis of plastics biodegradation process and developing more efficient methods for screening plastics biodegraders.

[Advances in methods for detecting plastics biodegradation].

Photoactivatable subcellular imaging offers the desirable spatiotemporal resolution in studying the complex biological processes. Developing proper and simple photoremovable protecting groups (PPGs) is the key for constructing such imaging probes. In this study, we designed and synthesized three Sanger's reagent caged fluorescent probes for specific photoactivated organelle imaging. We showed the photoactivated fluorescence recovery of the probes. The photolysis mechanism was verified by mass spectra (MS) analysis, and the highly efficient photolysis process was monitored by high performance liquid chromatography (HPLC). The probes successfully imaged their respective targeted organelle, including mitochondria, lysosomes, and endoplasmic reticulum (ER), in a photoactivated manner, and exhibited high co-localization coefficients when compared to commercially available probes. Given the high reactivity and efficient photodissociation capability of Sanger's reagent with amino groups, our examples demonstrate the potential of Sanger's reagent as a novel general PPG for constructing photoactivatable fluorescent probes. 

Sanger's reagent as a new general phototrigger for organelle imaging

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A Rhodol-Derived Probe for Intracellular Biothiols Imaging and Rapid Labelling of Sulfhydryl-Containing Proteins

Fluorescent Labeling of Dicysteine‐Tagged Peptide for Monitoring and Optimization of Protein Bio‐production in Bacteria

[Using (1)H-nuclear magnetic resonance metabolomics and gene ontology to establish pathological staging model for esophageal cancer patients].

[Advances in methods for detecting plastics biodegradation].

Sanger's reagent as a new general phototrigger for organelle imaging