Thienopyrimidine-derived multifunctional fluorescence sensor for the detection of Cu2+, Fe3+, and PPi in different solvents.

Hydrazine substituted thienopyrimidine, a new fluorophore, was used to synthesize a novel Schiff base R1 as a chemosensor via the condensation with p-formyltriphenylamine, and the structure was confirmed using nuclear magnetic resonance spectroscopy (NMR) and mass spectrometry (MS) analysis. When treated with Cu2+ in dimethylsulfoxide (DMSO)/H2O buffer, R1 showed a phenomenon of fluorescence quenching, which was reversible with the action of ethylenediaminetetraacetic acid (EDTA). When treated with Fe3+ in dimethylformamide (DMF)/H2O buffer, R1 exhibited the same phenomenon, but fluorescence was recovered with inorganic pyrophosphate (PPi) quantitatively. The complexation ratios for R1-Cu2+ and R1-Fe3+ were both 1:2, which were manifested by MS titrations and corresponding Job's plots. The limits of detection of R1 for Cu2+ and Fe3+ were 3.11 × 10-8 and 1.24 × 10-7 M, respectively. The sensing mechanism of R1 toward Cu2+ and Fe3+ was confirmed using density functional theory calculations and electrostatic potential analysis. Test strips of R1 were fabricated successfully for on-site detection of Cu2+ and Fe3+. In addition, R1 was applied to recognize Cu2+ and Fe3+ in actual water samples with satisfactory recovery.

Inositol pyrophosphate profiling reveals regulatory roles of IP6K2-dependent enhanced IP7 metabolism in the enteric nervous system.

Inositol pyrophosphates regulate diverse physiological processes; to better understand their functional roles, assessing their tissue-specific distribution is important. Here, we profiled inositol pyrophosphate levels in mammalian organs using an originally designed liquid chromatography-mass spectrometry (LC-MS) protocol and discovered that the gastrointestinal tract (GIT) contained the highest levels of diphosphoinositol pentakisphosphate (IP7) and its precursor inositol hexakisphosphate (IP6). Although their absolute levels in the GIT are diet dependent, elevated IP7 metabolism still exists under dietary regimens devoid of exogenous IP7. Of the major GIT cells, enteric neurons selectively express the IP7-synthesizing enzyme IP6K2. We found that IP6K2-knockout mice exhibited significantly impaired IP7 metabolism in the various organs including the proximal GIT. In addition, our LC-MS analysis displayed that genetic ablation of IP6K2 significantly impaired IP7 metabolism in the gut and duodenal muscularis externa containing myenteric plexus. Whole transcriptome analysis of duodenal muscularis externa further suggested that IP6K2 inhibition significantly altered expression levels of the gene sets associated with mature neurons, neural progenitor/stem cells, and glial cells, as well as of certain genes modulating neuronal differentiation and functioning, implying critical roles of the IP6K2-IP7 axis in developmental and functional regulation of the enteric nervous system. These results collectively reveal an unexpected role of mammalian IP7-a highly active IP6K2-IP7 pathway is conducive to the enteric nervous system.

Triazol-Methanaminium-Pillar[5]arene-Functionalized Single Nanochannel for Quantitative Analysis of Pyrophosphate in Water.

Inorganic pyrophosphate (PPi) is an important biological functional anion and plays crucial roles in life science, environmental science, medicine, and chemical process. Quantification of PPi in water has far-reaching significance for life exploration, disease diagnosis, and water pollution control. The label-free quantitative detection of PPi anions with a nanofluidic sensing device based on a conical single nanochannel is demonstrated. The channel surface is functionalized with a synthetic PPi receptor, triazol-methanaminium-functionalized pillar[5]arene (TAMAP5), using carbodiimide coupling chemistry. Due to the specific binding between TAMAP5 and PPi, the functionalized nanochannel can discriminate PPi from other inorganic anions with high selectivity through ionic current recording, even in the presence of various interfering anions. The current response exhibits a linear correlation with PPi concentration in the range from 1 × 10-7 to 1 × 10-4 M with a limit of detection of 6.8 × 10-7 M. A spike-and-recovery analysis of PPi in East Lake water samples indicates that the proposed nanofluidic sensor has the ability to quantitate micromolar concentrations of PPi in environmental water samples.

FeMoO4 nanospheres-based nanozymatic colorimetry for rapid and sensitive pyrophosphate detection.

Assays of pyrophosphate ion (PPi) are of remarkable biochemical significance due to their vital roles in the bioenergetic and metabolic processes or as disease indicators. Colorimetry is popular in the field of biosensing and detection because of its simplicity, speed and cost-effectiveness, but there is a lack of a suitable colorimetric probe. Herein, a novel colorimetric sensing platform has been established for the detection of pyrophosphate based on the FeMoO4-H2O2-3,3',5,5'-tetra-methylbenzidine (TMB) system. Compared with most previously reported iron-based nanozymes, the as-obtained FeMoO4 nanospheres with a rough surface possessed a much superior peroxidase-like catalytic activity (Vmax = 28.47 × 10-8 M s-1) and substrate affinity (Km = 0.174 mM) toward H2O2 catalysis. Due to the Fe(II) and PPi reaction, the presence of PPi could specifically restore blue oxidized TMB to colorless TMB, which led to a decrease in UV absorption at 652 nm. The absorbance change is proportional to the PPi concentration, with a linear detection range (from 0.5 to 25 µM) and a low detection limit of 0.3 µM (S/N = 3). Accordingly, its excellent selectivity and high sensitivity made it a potential colorimetric sensor for PPi analysis in actual water samples.

Portable, quantitative, and sequential monitoring of copper ions and pyrophosphate based on a DNAzyme-Fe3O4 nanosystem and glucometer readout.

In this report, portable, quantitative, and sequential monitoring of copper ions and pyrophosphate (PPi) with a single sensor based on a DNAzyme-Fe3O4 system and glucometer readout was performed. Initially, streptavidin was functionalized on the surface of magnetic Fe3O4 spheres through glutaraldehyde. Then, an invertase-modified DNA Cu substrate was connected to the magnetic Fe3O4 spheres by a specific reaction between streptavidin and biotin. The sensing system was formed by a hybridization reaction between the Cu substrate and Cu enzyme. In the presence of Cu2+, Cu2+ will recognize the Cu DNA substrate and form an "off-on" signal switch, thereby resulting in the separation of invertase from the Fe3O4 nanospheres. PPi recognizes Cu2+ to form a Cu2+-PPi complex, resulting in an "on-off" signal switch. Under optimized conditions, linear detection ranges for Cu2+ and PPi of 0.01-5 and 0.5-10 µM, and detection limits for Cu2+ and PPi of 10 nM and 500 nM, respectively, were obtained. Good selectivity was achieved for the analysis of Cu2+ and PPi. Satisfactory results were achieved for this biosensor during the determination of Cu2+ in real tap samples and PPi in human urine samples. This verified that the sensor is portable and low cost, and can be applied to the sequential monitoring of multiple analytes with a single point-of-care biosensor.

Plasmodium falciparum gametocyte-induced volatiles enhance attraction of Anopheles mosquitoes in the field.

BACKGROUND: Plasmodium parasites manipulate the interaction between their mosquito and human hosts. Patients infected with gametocytes attract anopheline mosquitoes differentially compared to healthy individuals, an effect associated with an increased release of attractive volatile cues. This odour-driven manipulation is partly mediated by the gametocyte-specific metabolite, (E)-4-hydroxy-3-methyl-but-2-enyl pyrophosphate (HMBPP), which induces increased release of select aldehydes and terpenes from red blood cells and results in the enhanced attraction of host-seeking mosquitoes, which are vectors of malaria. This study investigates the effect of the HMBPP-induced volatiles on the attraction of wild Anopheles mosquitoes to humans under field conditions. METHODS: The efficacy of the HMBPP-induced odour blend to attract Anopheles was evaluated in a 4 × 6 Latin rectangular study design indoors using baited Suna traps. Furthermore, to assess the efficacy of the HMBPP-induced odour blend in (1) augmenting the attractiveness of human odour, and (2) attracting Anopheles mosquitoes in the absence of human odour, a two-choice assay using host decoy traps (HDTs) was used and evaluated using binomial generalized regression. RESULTS: Traps baited with the HMBPP-induced odour blend attracted and caught both Anopheles arabiensis and Anopheles pharoensis females in a dose-dependent manner in the presence of background human odour, up to 2.5 times that of an unbaited trap. Given a choice between human odour and human odour laden with the HMBPP-induced odour blend, mosquitoes differentially preferred traps augmented with the HMBPP-induced odour blend, which caught twice as many female An. arabiensis. Traps baited with the HMBPP-induced odour blend but lacking the background of human odour were not effective in attracting and catching mosquitoes. CONCLUSION: The findings of the present study revealed that the HMBPP-induced odour blend, when augmented with human body odour, is attractive to anopheline mosquitoes and could be used as a complementary vector control tool along with existing strategies.

Effect of chlorine dioxide and phosphates on the quality of tiger frog (Rana tigrina) meat during 4 °C storage.

Tiger frog (Rana tigrina) meat is extremely perishable. This study investigated the antimicrobial efficacy of chlorine dioxide (ClO2 ) on frog meat, optimized the formulation of a phosphate-based enhancement solution by response surface methodology (RSM), and determined the quality parameters (i.e., total aerobic counts [TAC], pH, drip loss, cooking loss, color measurements, shear force, total volatile basic nitrogen [TVB-N], and thiobarbituric acid-reactive substances [TBARS]) of refrigerated frog meat pretreated with ClO2 and the optimized blend of phosphates. Treatments of frog meat with 35 and 70 ppm ClO2 for 3, 5, and 10 min achieved a 0.7-, 0.9- and 0.9-, and 0.8-, 1.4- and 1.6-log CFU/g reduction of TAC, respectively, indicating the antimicrobial efficacy of ClO2 was concentration- and time-dependent with such that higher concentrations and/or longer exposure time achieved greater bacterial reductions. The concentrations of the phosphates, including sodium tripolyphosphate (STPP), sodium pyrophosphate (SPP), and sodium hexametaphosphate (SHMP), were optimized as the formula of 0.3% STPP and 0.45% SPP obtaining the highest water retention of the frog meat. After washed with 70 ppm ClO2 for 10 min and subsequently soaked with 0.3% STPP and 0.45% SPP for 30 min, the frog meat stored at 4 °C shown significantly (P < 0.05) lower TAC (<4.4 log CFU/g) and higher water holding capacity during the whole storage of 12 days, compared to the control. Results indicated that the two-step process may be applicable to slow down deterioration and maintain quality frog meat during refrigeration. PRACTICAL APPLICATION: This research provides a means to slow down deterioration, maintain quality frog meat, and improve stability during refrigeration. Refrigerated frog meat products, which are preferred by consumers with juicier and more tender texture compared to the frozen-thawed meat, could be developed by the frog industry based on the data from this study.

Dual-emission carbon dots for ratiometric detection of Fe3+ ions and acid phosphatase.

We have developed a simple and convenient route to prepare fluorescent carbon dots with dual emission peaks respectively at 470 and 570 nm. The prepared dual-emission carbon dots can be used for ratiometric detection of Fe3+ ions in the range from 0 to 50 µmol·L-1 with 0.8 µmol·L-1 detection limit based on the fluorescence quenching at 570 nm. The quenched fluorescence induced by Fe3+ ions could be recovered by pyrophosphate. We further used the carbon dots-Fe3+ ions-pyrophosphate mixed system for ratiometric detection of acid phosphatase in the range from 0.08 to 6.75 µg·mL-1 with 0.01 µg·mL-1 detection limit.

Selective detection of pyrophosphate anion by zinc ensemble of C3-symmetric triaminoguanidine-pyrrole conjugate and its biosensing applications.

A new-fangled C3-symmetric triaminoguanidine-pyrrole conjugate has been constructed and utilized for sensing applications. The probe selectively detects zinc ions (Zn2+) by colorimetric as well as turn-on fluorescent manner. Further, the in-situ formed zinc ensemble displays turn-off fluorescence response towards the pyrophosphate anion (PPi) via displacement approach. Emissive off-on-off sensing characteristics of the probe has been successfully exploited to construct the INHIBIT logic gate, coding/decoding of messages and in vivo imaging of Zn2+/PPi in zebrafish larvae. Further, PPi detection characteristics of zinc ensembles were established for the sensing of PPi discharged from DNA synthesis and other biological reactions.

A novel fluorescent chemosensor based on coumarin and quinolinyl-benzothiazole for sequential recognition of Cu2+ and PPi and its applicability in live cell imaging.

In this study, a highly selective fluorescent sensor (E)-2-((2-(benzo[d]thiazol-2-yl)quinolin-8-yl)oxy)-N'-((7-(diethylamino)-2-oxo-2H-chromen-3-yl)methylene)acetohydrazide (TQC) was synthesized from 2-methylquinolin-8-ol and 4-(diethylamino)-2-hydroxybenzaldehyde and its structure was characterized by 1H NMR, 13C NMR, ESI-HR-MS and density functional theory (DFT) calculation. Sensor TQC showed an obvious "on-off-on" fluorescence response to Cu2+ and PPi in a DMSO/HEPES (3:2 v/v, pH = 7.4) buffer system. The detection limits of sensor TQC were 0.06 µM to Cu2+ and 0.01 µM to PPi. In addition, sensor TQC showed a 1:1 binding stoichiometry to Cu2+ and TQC-Cu2+ complex showed a 2:1 binding stoichiometry to PPi. The optimum pH range of sensor TQC and TQC-Cu2+ was 3-8. Further studies demonstrated that sensor TQC could be made into test paper strips for the qualitative of Cu2+ and PPi and showed sequentially "on-off-on" fluorescent bio-imaging of Cu2+ and PPi in HeLa cells.

Removal behaviors and mechanisms of orthophosphate and pyrophosphate by calcined dolomite with ferric chloride assistance.

Phosphate is one of the main contaminations in water, so an effective method of decreasing or removing phosphate is needed. The main purpose of this paper is to synthesize CaFe-LDHs and MgFe-LDHs from the mixture of calcined dolomite and ferric chloride to remove orthophosphate and pyrophosphate. The study showed that removal of orthophosphate was attributed to the precipitation by Ca2+ and adsorption by MgFe-LDHs, where the former played a main role. As for pyrophosphate, it was mainly removed by precipitation at the initial pyrophosphate concentration ranging from 3.228 to 17.04 mmol/L. When the initial concentrations became relatively higher, the removal efficiency of pyrophosphate decreased because the complexation effects by Fe3+, Ca2+ and Mg2+ took place.

Design of a nanoswitch for sequentially multi-species assay based on competitive interaction between DNA-templated fluorescent copper nanoparticles, Cr3+ and pyrophosphate and ALP.

The present work constructs a sequentially triggered nanoswitch (STN) for sequential detection of Cr3+, P2O74- (PPi) and alkaline phosphatase (ALP) depending on polythymine (T40) templated fluorescent Cu nanoparticles (Cu NPs). A significant phenomenon is that Cr3+ can only causing 5% QE of fluorescent Cu NPs synthesized by lower than 500 µM Cu2+, but the fluorescence of the Cu NPs synthesized by more than 500 µM Cu 2+ can be quenched up to 90% QE by the same concentration of Cr3+. Then the quenched fluorescence of CuNP-Cr3+ complex provides a sensing platform for PPi due to the strong binding between Cr3+ and PPi, resulting in dissociation of Cr3+ from the surface of Cu NPs and the recovery of fluorescence emission. Further ALP hydrolysis of PPi disrupts Cr3+-PPi assemble and Cr3+ is released to interact with Cu NPs, which induces fluorescence quenching again. Thus, sequentially detection of Cr3+ (LOD, 0.03 µM), PPi (LOD, 0.005 µM) and ALP (LOD, 0.125 mU/mL) was successfully implemented with high sensitivity and selectivity. The sensor is also successfully used for Cr3+, PPi and ALP assays in the human serum. Additionally, the sensitive "on-off-on-off" sensing behavior of the Cu NPs allow three chemical inputs (Cr3+, PPi and ALP) to construct a logic gate.

One-step synthesis of a dual-emitting carbon dot-based ratiometric fluorescent probe for the visual assay of Pb2+ and PPi and development of a paper sensor.

Herein, an easy and effective ratiometric fluorescent nanoprobe for the selective detection of Pb2+ and pyrophosphate (PPi) was developed based on label-free carbon dots (CDs). The CDs showed dual emission bands at 477 and 651 nm under a single-wavelength excitation (391 nm). Pb2+ and PPi were sequentially detected by the sensor with high reliability and accuracy via the exploitation of the fluorescence intensity ratio (F477/F651). The dual emission sensor exhibited high selectivity and excellent sensitivity for Pb2+ and PPi than for other analytes with detection limits as low as 0.055 µM and 0.089 µM, respectively. Moreover, the fluorescent CDs changed from pink to cyan and from cyan to pink and could be used as a fluorescent paper sensor for the visual detection of Pb2+ and PPi. To the best of our knowledge, this is the first report on the ratiometric fluorescence detection of Pb2+ and PPi by CDs; therefore, the as-synthesized CDs show great potential for application in the determination of Pb2+ and PPi in real samples.

Smart probe for simultaneous detection of copper ion, pyrophosphate, and alkaline phosphatase in vitro and in clinical samples.

Wilson's disease (WD), which might lead to acute liver failure, is an inherited disorder characterized by accumulation of copper (Cu2+) in the brain, the liver, and other vital organs. In the clinic, decreased serum alkaline phosphatase (ALP) concentration is used for WD diagnosis. But to the best of our knowledge, using a fluorescent probe to simultaneously detect multiple factors in WD (e.g., Cu2+, pyrophosphate (PPi), and ALP) has not been reported. Herein, we rationally designed a fluorescent switch (E)-8-((4-methylbenzylidene)amino)napthalen-1-amine (L) and successfully applied it for sequential and selective detections of Cu2+, PPi, and ALP in vitro, in living cells and synovial fluid samples with "Off," "On," and "Off" fluorescence signals, respectively. Considering the obvious correlations among Cu2+, PPi, and ALP in WD, we envision that our fluorescent probe L could be applied to in vitro diagnosing WD in the near future.

Signal-on fluorescence assay for pyrophosphate ions based on DNA-stabilized silver nanoclusters.

Pyrophosphate anion (P2 O7 4- , PPi) is considered as a potential biomarker for arthritic diseases because high levels of PPi may result in calcium pyrophosphate dehydrate crystal deposition diseases. In this study, a simple fluorescence method for PPi was demonstrated by organic integration of the efficient fluorescence quenching ability of copper ions to DNA-scaffolded silver nanoclusters and the strong affinity of PPi towards copper ions. This simple fluorescence sensor showed a low detection limit (0.28 µM based on signal/noise = 3) towards the detection of PPi. Practical application of this method was also validated by detection of PPi in the synovial fluid.

Polyphosphate Chain Length Determination in the Range of Two to Several Hundred P-Subunits with a New Enzyme Assay and 31P NMR.

Currently, 31P NMR is the only analytical method that quantitatively determines the average chain length of long inorganic polyphosphate (>80 P-subunits). In this study, an enzyme assay is presented that determines the average chain length of polyphosphate in the range of two to several hundred P-subunits. In the enzyme assay, the average polyP chain length is calculated by dividing the total polyphosphate concentration by the concentration of the polyphosphate chains. The total polyphosphate is determined by enzymatic polyphosphate hydrolysis with Saccharomyces cerevisiae exopolyphosphatase 1 and S. cerevisiae inorganic pyrophosphatase 1, followed by colorimetric orthophosphate detection. Because the exopolyphosphatase leaves one pyrophosphate per polyphosphate chain, the polyphosphate chain concentration is assayed by coupling the enzymes exopolyphosphatase (polyP into pyrophosphate), ATP sulfurylase (pyrophosphate into ATP), hexokinase (ATP into glucose 6-phosphate), and glucose 6-phosphate dehydrogenase (glucose 6-phosphate into NADPH), followed by fluorometric NADPH detection. The ability of 31P NMR and the enzyme assay to size polyP was demonstrated with polyP lengths in the range from 2 to ca. 280 P-subunits (no polyP with a longer chain length was available). The small deviation between methods (-4 ± 4%) indicated that the new enzyme assay performed accurately. The limitations of 31P NMR (i.e., low throughput, high sample concentration, expensive instrument) are overcome by the enzyme assay that is presented here, which allows for high sample throughput and requires only a commonly available plate reader and micromole per liter concentrations of polyphosphate.

Spectrophotometric, fluorimetric and electrochemical selective pyrophosphate/ATP sensing based on the dimethyltin(IV)-tiron system.

Sensing of pyrophosphate anion (PPi) in the presence of nucleotide triphosphates allows the real time monitoring of the polymerase chain reaction. To get a deeper understanding of the factors involved in PPi/nucleotide triphosphate discrimination, a detailed study on the performance of a dimethyltin (IV)-catecholate complex capable of both separate fluorimetric or electrochemical detection of PPi in the presence of adenosine triphosphate (ATP) has been undertaken. Dimethyltin (IV) tightly binds PPi or ATP, and forms a stable 1:1 complex with tiron (4,5-dihydroxy-1,3-benzenedisulfonic acid) in water. The complexation equilibria with all components are characterized quantitatively by potentiometric and spectroscopic titrations. Pyrophosphate anion can be detected owing to its ability to release free tiron from the complex by measuring either a fluorimetric or an electrochemical signal. On the contrary, ATP does not displace tiron but causes an interference with PPi in the fluorimetric detection method due to the formation of a ternary Me2Sn(IV)-tiron-ATP complex with optical properties intermediate between those of free and bound tiron. In the electrochemical (square wave voltammetry) method, the ternary ATP complex shows a separate peak which does not coincide with the peaks of neither free nor bound tiron, thus making possible the simultaneous detection of ATP in addition to PPi.

In situ formation of fluorescent polydopamine catalyzed by peroxidase-mimicking FeCo-LDH for pyrophosphate ion and pyrophosphatase activity detection.

As pyrophosphate ion (PPi) and pyrophosphatase (PPase) play crucial roles in the pathological process of arthritis, determination of PPi and PPase in biological fluids turns to be of great importance for clinical diagnosis and therapy of arthritic diseases. In this work, we proposed a new fluorescent assay for PPi and PPase activity detection based on the competitive coordination chemistry of Fe3+ between PPi and an in situ formed fluorescent polydopamine (PDA). FeCo layered double hydroxide (FeCo-LDH) was explored as a peroxidase mimic to facilitate the in situ formation of fluorescent PDA from dopamine mediated by low-concentration H2O2 within 30 min; The formed fluorescent PDA could be significantly quenched by Fe3+ through forming a PDA-Fe3+ complex structure; When PPi existed, it coordinated Fe3+ competitively against PDA and inhibited the fluorescence quenching of PDA by Fe3+; When PPi was hydrolyzed under the catalysis of PPase, the Fe3+ ion could quench the fluorescence of the formed PDA again. With these principles, our fluorescent assay was able to detect PPi and PPase activity specifically, providing detection limits down to 54 µM and 0.13 U/L, respectively. Furthermore, accurate determination of PPi and PPase activity in spiked human serum was also demonstrated using the developed assay.

A Conjugated Polymer Fluorescent Sensor for Continuous Identification of Copper(II) and Pyrophosphate in Blood Serum and Synovial Fluid.

A novel "on-off-on" super-sensitive conjugated polymer fluorescence sensor (PPE-DPA) was developed and it was applied to realize the continuous recognition of Cu2+ and pyrophosphate (PPi). The fluorescence intensity decreased linearly with the change of Cu2+ from 0.05 to 5.0 µmol L-1 and the limit of detection was 24 nmol L-1. The fluorescence intensity was linearly enhanced with the increase of PPi from 0.5 to 12.0 µmol L-1 and the limit of detection was 230 nmol L-1. In addition, this method was applied to detect PPi in the blood serum and synovial fluid of patients with arthritis and satisfactory results were obtained. Thus, the PPE-DPA is not only an effective tool for detecting Cu2+ and PPi in samples, but also presents a potential way to diagnose arthritis.

A water-soluble benzoxazole-based probe: Real-time monitoring PPi via situ reaction by two-photon cells imaging.

Pyrophosphate (PPi) played crucial roles in various fundamental physiological processes. Herein, a two-photon absorption (TPA) "On-Off-On" type benzoxazole-based fluorescence probe BN was designed and synthesized, which detected PPi through Cu2+ displacing method in situ system in aqueous medium. The on-off-on process of BN recognizing PPi was verified by mass spectra and theoretical calculations, which was successfully applied in TPA cells imaging.