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The development of portable, cost-effective, and straightforward DNA biosensors holds immense importance in various fields, including healthcare, environmental monitoring, and food safety. This study contributes to the objective by introducing an innovative approach for synthesizing carbon dots (Cdots) with high quantum yield (QY) and remarkable selectivity for Fe 3+ ions. Utilizing o-phenylenediamine as a precursor, the study achieved a straightforward and environmentally friendly synthesis method, enabling the efficient detachment of metal ions from the Cdot surface upon introducing pyrophosphate (PPi). The presence of surface hydroxyl and amino groups facilitated specific Fe 3+ recognition. Employing D-optimal response surface methodology, the study optimized Cdot synthesis parameters, identifying temperature and heating time as critical factors influencing QY. Statistical analysis confirmed the model's reliability, predicting maximum QY of 48.8 % with minimal deviation from experimental results. Characterization studies revealed the amorphous nature of Cdots through HR-TEM, XRD, and FTIR analysis. Furthermore, the proposed LAMP/PPi biosensing technique demonstrated higher sensitivity, specificity, and repeatability, with negligible interference from common anions and efficacy across varying pH levels. The limit of detection (LOD) of 0.079 (±0.01) μM and the detection range of 0.1 μM-2 mM underscore the biosensor's practical utility. This study highlights a promising direction for developing paper-based LAMP/PPi biosensors with potential diagnostics and environmental monitoring applications. Significantly, the biosensing technique is applicable to any DNA amplification method generating pyrophosphate (PPi) as a by-product., Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: Dinesh Prasad reports financial support was provided by Birla Institute of Technology. DINESH PRASAD reports a relationship with Birla Institute of Technology., (Copyright © 2024 Elsevier Inc. All rights reserved.)

The residues of organophosphorus pesticides (OPs) are increasing environmental pollution and public health concerns. Thus, the development of simple, convenient and sensitive method for detection of OPs is crucial. Herein, a multifunctional Fe-based MOF with fluorescence, catalytic and adsorption, is synthesized by a simple one-pot hydrothermal method. The ratiometric fluorescence sensor for detection of OPs is constructed by using only one multifunctional sensing material. The NH 2 -MIL-101(Fe) is able catalyze the o-phenylenediamine (OPD) into 2,3-diaminophenazine (DAP) in the presence of H 2 O 2 . The generated DAP can significantly quench the intrinsic fluorescence of NH 2 -MIL-101(Fe) by the fluorescence resonance energy transfer (FRET) and internal filtration effect (IFE), while producing a new measurable fluorescence. Without immobilization or molecular imprinting, pyrophosphate ion (PPi) can inhibit the peroxidase-like activity of the NH 2 -MIL-101(Fe) by chelating with Fe 3+ /Fe 2+ redox couple. Moreover, PPi can also be hydrolyzed by alkaline phosphatase (ALP), the presence of OPs inhibits the activity of ALP, resulting in the increase of extra PPi preservation and signal changes of ratiometric fluorescence, the interactions of ALP with different OPs are explored by molecular docking, the OPs (e.g., glyphosate) interact with crucial amino acid residues (Asp, Ser, Ala, Lys and Arg) are indicated. The proposed sensor exhibits excellent detection performance for OPs with the detection limit of 18.7 nM, which provides a promising strategy for detection of OPs., Competing Interests: Declaration of competing interest The authors have no conflict of potential competing interest, financial or otherwise., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Sensitive and accurate analysis of pyrophosphate (PPi) is of great importance for preventing health hazard in environment. Nevertheless, most of sensors focus on sensitivity and selectivity, but practicality is also a significant quota. How to reconciling sensitivity, selectivity and practicability in one single sensor is desirable but remains challenging. Here, we created a novel metal-carbon nanozyme V 2 O 5 @C with two-dimensional (2D) morphology and high yet exclusive peroxidase (POD)-like activity via a glucose and NH 4 NO 4 -co-directed avenue, and further showed its application in constructing a portable and disposable paper-based analytical chip (PA-chip) for rapid, visual and onsite analysis of PPi. PPi etched V 2 O 5 to prevent the decomposition of H 2 O 2 into ·OH, resulting in weakened POD-like activity. In comparison with PPi deficiency, colorless TMB couldn't be oxidized into oxidized TMB with a dropped absorption at 652 nm. Therefore, obviously shallowed blue color on PA-chip surface was recorded, and demonstrated a negative relationship with PPi dosage, enabling rapid and visual detection of PPi with a limit of detection of 2.6 nM. This study demonstrated the burgeoning applications of nanozymes with POD-like activity in construction of PA-chips for PPi and will quicken the advancement of practical sensors, guaranteeing environmental safety., Competing Interests: Declaration of competing interest The authors declare no competing financial interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Elucidation on the emulsifying behaviors of goose liver protein (GLP) from interfacial perspective was scarce when protein charging was altered. This work aimed to elucidate the role of phosphorylation on the interfacial associative interaction and then emulsion stabilizing properties of GLP using three structurally relevant phosphates of sodium trimetaphosphate (STMP), sodium tripolyphosphate (STPP) and sodium pyrophosphate (TSPP). A monotonic increment of protein charging treated from STMP, STPP to TSPP caused progressively increased particle de-aggregation, surface hydrophobicity and structural flexibility of GLP. Compared with STMP and TSPP, STPP phosphorylation rendered the most strengthened interfacial equilibrium pressure (11.98 ± 0.24 mN/m) due to sufficient unfolding but moderated charging character conveyed. Desorption curve and interfacial protein microstructure indicated that STPP phosphorylation caused the highest interfacial connectivity between proteins adsorbed onto the same droplet, as was also verified by interfacial elastic modulus (10.3 ± 0.21 mN/m). STPP treated GLP also yielded lowest droplet size (8.16 ± 0.10 μm), flocculation (8.18%) and Turbiscan stability index (8.78 ± 0.36) of emulsion but most improved microrheological properties. Overall, phosphorylation functioned itself in fortifying the intradroplet protein-protein interaction but restraining the interdroplet aggregation, and STPP phosphorylation endowed the protein with most enhanced interfacial stabilization and emulsifying efficiency., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

T4 polynucleotide kinase (T4 PNK) phosphorylates the 5'-terminus of DNA and RNA substrates. It is widely used in molecular biology. Single nucleotides can serve as substrates if a 3'-phosphate group is present. In this study, the T4 PNK-catalyzed conversion of adenosine 3'-monophosphate (3'-AMP) to adenosine-3',5'-bisphosphate was characterized using isothermal titration calorimetry (ITC). Although ITC is typically used to study ligand binding, in this case the instrument was used to evaluate enzyme kinetics by monitoring the heat production due to reaction enthalpy. The reaction was initiated with a single injection of 3'-AMP substrate into the sample cell containing T4 PNK and ATP at pH 7.6 and 30 °C, and Michaelis-Menten analysis was performed on the reaction rates derived from the plot of differential power versus time. The Michaelis-Menten constant, K M , was 13 μM, and the turnover number, k cat , was 8 s -1 . The effect of inhibitors was investigated using pyrophosphate (PP i ). PP i caused a dose-dependent decrease in the apparent k cat and increase in the apparent K M under the conditions tested. Additionally, the intrinsic reaction enthalpy and the activation energy of the T4 PNK-catalyzed phosphorylation of 3'-AMP were determined to be -25 kJ/mol and 43 kJ/mol, respectively. ITC is seldom used as a tool to study enzyme kinetics, particularly for technically-challenging enzymes such as kinases. This study demonstrates that quantitative analysis of kinase activity can be amenable to the ITC single injection approach., Competing Interests: Declaration of competing interest The authors declare that they have no conflicts of interest with the contents of this article., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Background: Inorganic pyrophosphatase (PPase) is key enzyme playing a key role in biochemical transformations such as biosynthesis of DNA and RNA, bone formation, metabolic pathways associated with lipid, carbohydrate and phosphorous. It has been reported that lung adenocarcinomas, colorectal cancer, and hyperthyroidism disorders can result from abnormal level of PPase. Therefore, it is of notable significance to develop simple and effective real time assay for PPase enzyme activity monitoring for screening of many metabolic pathways as well as for early disease diagnosis., Result: The fluorometric detection of PPase enzyme in near infrared region-1 (NIR-1) has been carried out using bimetallic nanoclusters (LA@AuAg NCs). The developed sensing strategy was based on quenching of fluorescence intensity of LA@AuAg NCs upon interaction with copper (Cu 2+ ) ions. The off state of LA@AuAg_Cu 2+ ensemble was turned on upon addition of pyrophosphate anion (PPi) due to strong binding interaction between PPi and Cu 2+ . The catalytic conversion of PPi into phosphate anion (Pi) in the presence of PPase led to liberation of Cu 2+ ions, and again quenched off state was retrieved due to interaction of free Cu 2+ with LA@AuAg NCs. The ultrasensitive detection of PPase was observed in the linear range of 0.06-250 mU/mL with LOD as 0.0025 mU/mL. The designed scheme showed good selectivity towards PPase enzyme in comparison to other bio-substrates, along with good percentage recovery for PPase detection in real human serum samples., Significance: The developed NIR based assay is ultrasensitive, highly selective and robust for PPase enzyme and can be safely employed for other enzymes detection. This highly sensitive nature of biosensor was result of involvement of fluorescence-based technique and synergistic effect of dual metal in NIR based bimetallic NCs. Moreover, owing to the emission in NIR domain, in future, these nanoclusters can be safely employed for many biomedical applications for In vivo studies., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

It is a challenging task to remove heavy metal ions efficiently from the wastewater containing high concentrations of complexants. In this work, a novel PEI-modified chitosan/activated carbon composite adsorbent (PCA) was prepared and applied to the removal of Cu(II) from pyrophosphate plating rinsing wastewater. The main species of Cu(II) in the pyrophosphate wastewater was [Cu(HP 2 O 7 ) 2 ] 4- or [Cu(P 2 O 7 ) 2 ] 6- , which were denoted as [Cu(II)-PP] anions. The maximum adsorption capacity of PCA for Cu(II) reached 1.41 mmol g -1 under the condition of pH = 8 and molar ratio of pyrophosphate to Cu(II) = 4:1. The adsorption kinetic behavior of Cu(II) on PCA followed the Elovich model best and PCA attained adsorption equilibrium within 36 h. The thermodynamic studies showed that the adsorption process of Cu(II) by PCA was endothermic and spontaneous. The PCA fixed bed column was used to remove Cu(II) from simulated pyrophosphate plating rinsing wastewater. After three consecutive adsorption-desorption cycles, the adsorption performance, hydraulic conductivity, and mechanical stability of PCA column did not decrease. The FTIR and XPS analysis results indicated that [Cu(II)-PP] anions can be adsorbed on PCA by electrostatic attraction with protonated amine groups or coordination with the amine groups of PCA via ligand substitution., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Protein O-linked β-D-N-acetylglucosamine (O-GlcNAc) modification (O-GlcNAcylation), an essential post-translational as well as cotranslational modification, is the attachment of β-D-N-acetylglucosamine to serine and threonine residues of nucleocytoplasmic proteins. An aberrant O-GlcNAc profile on certain proteins has been implicated in metabolic diseases such as diabetes and cancer. Inhibitors of O-GlcNAc transferase (OGT) are valuable tools to study the cell biology of protein O-GlcNAc modification. In this study we report novel uridine-peptide conjugate molecules composed of an acceptor peptide covalently linked to a catalytically inactive donor substrate analogue that bears a pyrophosphate bioisostere and explore their inhibitory activities against OGT by a radioactive hOGT assay. Further, we investigate the structural basis of their activities via molecular modelling, explaining their lack of potency towards OGT inhibition., (Copyright © 2021 Elsevier Inc. All rights reserved.)

The utility of plasma-treated winter mushroom powder (PWMP) as an alternative to synthetic nitrite and phosphate in ground ham was evaluated. Treatment of atmospheric non-thermal plasma generated nitrite in winter mushroom homogenate, and PWMP contained 4.87 g/kg of nitrite. Canned ground ham samples without nitrite and phosphate (NC), with sodium nitrite and sodium pyrophosphate (PC), and with PWMP (PWM) were manufactured, sterilized (F 0  > 4), and stored under accelerated conditions (40 °C) for 30 days. The nitrosyl-hemochrome content and color of PWM were similar to those of PC, without differences in the thiobarbituric acid reactive substances (TBARS) value. The proportion of jelly and melted fat exuded from ground hams in PWM exceeded that in PC but was lower than that in NC (P < .05). Therefore, PWMP can successfully replace synthetic nitrite, but it is an insufficient substitute for phosphate in ground ham., Competing Interests: Declaration of interests The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Magnetic trapping has been employed in the development of analytical methods owing to its ease and simplicity in handling samples. Nevertheless, the generation of functional probes is usually time consuming. A new and simple affinity method that uses gadolinium ion (Gd 3+ ), a magnetic ion, as affinity probe for magnetic tapping of pathogenic bacteria was demonstrated in the present study. Escherichia coli O157:H7, Staphylococcus aureus, and Acinetobacter baumannii were selected as model bacteria. The model bacteria were magnetically isolated after incubation in Tris buffer (pH 8) containing Gd 3+ (0.1 M) under microwave heating (power: 180 W, 90 s × 3). The resultant Gd 3+ -bacterium conjugates possessed sufficient magnetism, resulting in magnetic aggregations by an external magnet (∼4,000 Gauss). For ease of magnetic isolation, the sample containing Gd 3+ -bacterium complexes was stirred by a small magnet. After 1 h, the magnet attached with precipitates, i.e., Gd 3+ -bacterium conjugates, was readily removed using a pair of tweezers. The bacteria in the resultant conjugates were characterized by matrix-assisted laser desorption/ionization mass spectrometry. The limits of detection of the current approach toward E. coli O157:H7, S. aureus, and A. baumannii in complex samples were ∼10 4 -10 5  cells mL -1 ., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020. Published by Elsevier B.V.)

Background: Bouillon cubes are a potential vehicle for iron fortification. They are currently fortified with ferric pyrophosphate (FePP), which is known to be poorly absorbed. The objective of this study was to assess the iron absorption of Aspergillus oryzae grown in FePP (ASP-p) and compare it with FePP and ferrous sulfate (FeSO4)-fortified bouillon cubes., Methods: In 2 single-blinded, crossover studies, healthy women with serum ferritin concentrations <40 μg/L were randomly assigned to consume a rice-vegetable meal with iron-fortified chicken bouillon. Subjects in study I (n = 17, 18-26 y) consumed iron from both iron sources as 57FePP and 58ASP-p (intrinsically labeled with 58FePP) with a meal containing 4.2 mg of total iron provided for 3 d. Study II (n = 18, 18-29 y) was similar except that subjects consumed 57FeSO4 and 58ASP-p. Whole-blood stable isotope enrichment after 14 d was used to measure fractional iron absorption. Hemoglobin, hematocrit, serum ferritin, hepcidin, and serum C-reactive protein were analyzed at baseline and at 14 d. A t test was used to compare the mean differences in fractional absorptions within each study and baseline characteristics between studies., Results: Geometric mean (95% CI) fractional iron absorption of FePP [0.94% (0.63%, 1.40%)] was lower than ASP-p [2.20% (1.47%, 3.30%)] (P < 0.0001) in study I. In study II, ASP-p fractional absorption [2.98% (2.03%, 4.38%)] was lower than that of FeSO4 [9.88% (6.70%, 14.59%)] (P < 0.0001). Both ferritin (r = -0.41, P = 0.014) and hepcidin (r = -0.42, P = 0.01) concentrations were inversely correlated with ASP-p iron absorption. Fractional absorption of ASP-p was also positively correlated with FePP (r = 0.92, P < 0.0001) and FeSO4 (r = 0.52, P < 0.02) absorption., Conclusions: ASP-p-fortified bouillon provided 2.3-fold higher absorbable iron than the currently used FePP. Bouillon fortified with ASP-p may contribute sufficient bioavailable iron to meet the daily iron requirements in young women only if consumed with other iron-fortified staple foods. This trial was registered at clinicaltrials.gov as NCT03586245., (Copyright © The Author(s) 2020.)

In this study we applied the dual-responsive chromo-fluorescent Cu 2+ chelate 1C for the recognition of miRNA-146a through a pyrophosphate (PPi) sensing strategy in a rolling circle amplification (RCA) process. This approach for the recognition of miRNA-146a was highly robust, selective, and sensitive down to the attomolar (fluorogenic) and sub-micromolar (chromogenic) ranges under modified biochemical conditions at elevated temperature. Probe 1 selectively recognized Cu 2+ and PPi ions in a sequential manner, as evidenced by colorless→pink→colorless transitions; the fluorescence emissions centered at 480 nm underwent a corresponding on-off-on sequence in the bluish-green region. We attribute this reversible switching upon the addition of Cu 2+ /PPi ions to effective chelation-induced ligand-to-metal charge/electron transfer that resulted in opening of the lactam ring upon complexation and closing of the lactam ring upon decomplexation. We also report a label-free approach for monitoring miRNA-146a amplification in an RCA process under modified T4 ligase and ϕ 29 buffer conditions, using the Cu 2+ ensemble 1C at pH 7.0 (4-(2-hydroxyethyl)piperazine-1-ethanesulfonic acid: HEPES, 10 mM MgCl 2 ); the time required to perform this process (40-50 min) was relatively shorter than conventional RCA process. This ensemble 1C could recognize miRNA-146a colorimetrically (from pink to colorless) and fluorimetrically ("turn-on" mode) at concentrations within the highly sensitive atto-/nanomolar range under physiological conditions. This cost-effective label-free sensing strategy appears to be a universal method for detecting miRNAs according to the specified length of the template., Competing Interests: Declaration of competing interest Authors declare no conflicts of interest., (Copyright © 2019 Elsevier Inc. All rights reserved.)

The present work constructs a sequentially triggered nanoswitch (STN) for sequential detection of Cr 3+ , P 2 O 7 4- (PPi) and alkaline phosphatase (ALP) depending on polythymine (T40) templated fluorescent Cu nanoparticles (Cu NPs). A significant phenomenon is that Cr 3+ can only causing 5% QE of fluorescent Cu NPs synthesized by lower than 500 μM Cu 2+ , 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 Cr 3+ . Then the quenched fluorescence of CuNP-Cr 3+ complex provides a sensing platform for PPi due to the strong binding between Cr 3+ and PPi, resulting in dissociation of Cr 3+ from the surface of Cu NPs and the recovery of fluorescence emission. Further ALP hydrolysis of PPi disrupts Cr 3+ -PPi assemble and Cr 3+ is released to interact with Cu NPs, which induces fluorescence quenching again. Thus, sequentially detection of Cr 3+ (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 Cr 3+ , 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 (Cr 3+ , PPi and ALP) to construct a logic gate., (Copyright © 2019 Elsevier B.V. All rights reserved.)

As an important biomarker, alkaline phosphatase (ALP) is one of the most commonly assayed enzymes in clinical practice. Here a novel turn-on fluorescent nanoswitch for ALP assay was suggested. The nanoswitch was easily constructed via two high-affinity ligands between GSH and Al 3+ and PPi and Al 3+ based on the difference in the affinity. The primary fluorescence of as-prepared GSH capped Cu nanoclusters (NCs) turned on first upon the Al 3+ addition due to the Al 3+ induced aggregation induced emission (AIE) enhancement based on the high affinity of GSH and Al 3+ . The presence of PPi then made Al 3+ desorb from the surface of GSH capped Cu NCs due to the higher affinity of PPi and Al 3+ . As a result, the fluorescence of the Cu NCs was quenched. ALP could hydrolyze PPi into phosphate, destroying the PPi-Al 3+ complex and releasing Al 3+ . Thus, the Al 3+ binds to GSH again and the fluorescence was restored. The nanoswitch was demonstrated to be sensitive and selective for ALP assay and was successfully used for the ALP assay in the human serum., (Copyright © 2019 Elsevier B.V. All rights reserved.)

For the first time, a biomass-derived porous carbon cathode (WDC) was fabricated via a facile one-step pyrolysis of recovered wood-waste without any post-treatment. The WDC along with pyrophosphate (PP) as electrolyte were used in electro-Fenton (EF) at pH 8 for sulfathiazole (STZ) treatment. The H 2 O 2 accumulation capacity of WDC was optimized via the following parameters: pyrolysis temperature, applied current and electrolyte. Results showed that the WDC cathode prepared at 900 °C achieved the highest H 2 O 2 accumulation (13.80 mg L -1 in 3 h) due to its larger electroactive surface area (28.81 cm 2 ). Interestingly, it was found that PP decreased the decomposition rate of H 2 O 2 in solution as compared to conventional electrolyte, which resulted in higher H 2 O 2 accumulation. PP allowed operating EF at pH of 8 due to the formation of Fe 2+ -PP complexes in solution. Moreover, Fe 2+ -PP was able to activate oxygen to produce OH. In this way, the degradation of STZ took place through four main pathways: 1) via OH from the Fe 2+ -PP complex, 2) via OH from EF reactions, 3) via surface OH at the boron doped diamond electrode (BDD) and 4) via SO 4 - from BDD activation. Finally, microtox tests revealed that some toxic intermediates were generated during WDC/BDD/PP EF treatment, but they were removed at the end of the process., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 Me 2 Sn(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., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Chemical pretreatment can reduce NH 3 -N levels in piggery wastewater to a certain extent, but the lack of a carbon source for subsequent biological treatment leads to a low denitrification efficiency and poor total nitrogen (TN) removal percentage. Taking superphosphate (SP) pretreatment (SP/Pretreatment) as the control, this research studies the influence of fermented superphosphate (FSP; SP plus a carbon source) pretreatment (FSP/Pretreatment) on biological denitrification in a subsequent biological treatment step. Furthermore, the removal of pollutants under different influent modes is also evaluated. The experimental results show that with the addition of the SP pretreatment, the removal percentage of NH 3 -N was 52%, and the chemical oxygen demand (COD)/TN ratio increased from 0.36 to 0.71. However, with the addition of the FSP pretreatment, the removal percentage of NH 3 -N reached 64%, and the COD/TN ratio increased to 2.28. The combination of the FSP pretreatment and a subsequent sequencing batch reactor (SBR) step in the step-feed influent mode resulted in the best denitrification, with a TN removal percentage of 57%. This result was 51% higher than that of the SP/Pretreatment-SBR system, indicating that the addition of the FSP pretreatment improves the biological denitrification of piggery wastewater. After the full treatment process for piggery wastewater, the effluent COD was 57.33 mg·L -1 , the NH 3 -N was 66.32 mg·L -1 , and the total phosphorus (TP) was 1.17 mg·L -1 , all of which meet the emission standards of the "Fouling Standards for Pollutants in the Livestock Breeding Industry" (GB 18596-2001) (400 mg·L -1 COD, 80 mg·L -1 NH 3 -N, 8 mg·L -1 TP)., (Copyright © 2019 Elsevier B.V. All rights reserved.)

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 Cu 2+ 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., (Copyright © 2018 Elsevier B.V. All rights reserved.)

The objective was to examine shelf stability, cooked product yield, and sensory characteristics of beef patties that had no binder (Control), incorporated soy flour (Textured Vegetable Protein; TVP) or one of three dry potato extracts: X-TRATOS™ (potato extract), X-TRATOS™ O (potato extract with mustard), or X-TRATOS™ W (potato extract with sodium acid pyrophosphate). In retail display patties, all binders decreased discoloration and lipid oxidation compared to Control, and X-TRATOS™ O was superior (P < 0.05) to all other treatments. Cooking yield was higher (P < 0.05) in patties containing potato extracts compared with patties containing TVP, which had higher yield than Control patties. Beef patties with potato extracts were juicier (P < 0.05) than Control and TVP patties and had higher (P < 0.05) overall acceptability than Control patties. We conclude that potato extracts are effective binders for use in fresh or precooked beef patties because they improve retail shelf life, cooked product yield, and sensory characteristics., (Copyright © 2018. Published by Elsevier Ltd.)

Single superphosphate (SSP) as an additive could improve phosphorus availability and reduce nitrogen loss for composts, but few studies have explored the influence of SSP on the transformation of carbon fractions in composting. The aim of this work was to assess the effect of different components of SSP, including calcium dihydrogen phosphate (CDP), calcium sulfate (CS) and free acid (FA) on organic matter degradation and maturity during pig manure composting. The results showed that CDP had significantly negative effects on the duration of thermophilic phase and organic matter degradation, but lengthened the curing phase for the transformation of organic matter. FA could intensify the inhibiting effect of CDP and postpone the biodegradation process of composting, but CS could buffer the effect of CDP on the degradation of organic carbon fractions by controlling pH. The study reveals the roles of different components of SSP to the transformation of organic carbon fractions, which lays a foundation for regulating the effects of chemical additives during composting. Regulating the content of CDP in SSP or applying SSP with other chemical additives to control the biotoxicity of excess phosphate on microbial activity should be concerned for complete and efficient composting in further study., (Copyright © 2018. Published by Elsevier B.V.)

Nanozymes have been extensively exploited in the construction of colorimetric sensors in the past decade. However, as lack of specific recognition element, these sensors are mostly limited in the detection of hydrogen peroxide and its related agents. In this work, a colorimetric assay for the detection of alkaline phosphatase (ALP) activity was developed by employing copper (II)-based metal-organic frameworks (Cu-MOFs) as peroxidase mimic and pyrophosphate (PPi) as recognition element. Benefiting from its intrinsic porosity, Cu-MOFs built from one-step self-assembly showed a superior catalytic activity over horseradish peroxidase. It was found that, in the presence of PPi, the catalytic activity of Cu-MOFs was greatly inhibited due to the ultrastrong binding ability of Cu 2+ with PPi, and no colorimetric signal was recorded. However, after incubation with ALP, such negative behavior of PPi can be suppressed as the hydrolysis of PPi into orthophosphates (Pi) with low affinity to Cu 2+ , producing a colorimetric signal depending on ALP level. On this basis, a way for quantitative analysis of ALP activity was paved accordingly. Compared with previously reported ALP assays based on nanomaterials, this assay is simple, cost-effective and visualized, which not only allows highly sensitive analysis of ALP activity with a detection limit of 0.19 U/L, but also exhibits a good sensing performance in serum samples. Furthermore, based on the distinct colorimetric signals induced by PPi and ALP, an IMPLICATION logic gate was constructed. We hope that this study could provide a new insight for designing nanozyme-based highly specific colorimetric sensors., (Copyright © 2017 Elsevier B.V. All rights reserved.)

The mode of interaction and thermodynamic stability of complexes formed in binary and ternary Cu(II)/ATP/triamines systems were studied using potentiometric and spectroscopic (NMR, EPR, UV-Vis) methods. It was found that in binary metal-free systems ATP/H x PA species are formed (PA: Spd=spermidine or 3,3-tri=1,7-diamino-4-azaheptane) where the phosphate groups from nucleotides are preferred negative centers and protonated amine groups of amines are positive centers of reaction. In the ternary systems Cu/ATP/H x (PA) as well as Cu/(ATP)(PA) species are formed. The type of the formed Cu(II) complexes depends on pH of the solution. For a low pH value the complexation appears between Cu(II) and ATP molecules via oxygen atoms of phosphate groups. For a very high pH value, where ATP is hydrolyzed, the Cu(II) ions are bound to the nitrogen atoms of polyamine molecules. We did not detect any direct coordination of the N7 nitrogen atom of adenosine to Cu(II) ions. It means that the CuN7 interaction is an indirect type and can be due to noncovalent interplay including water molecule. EPR studies were performed at glassy state (77K) after a fast freezing both for binary and ternary systems. The glassy state EPR spectra do not reflect species identified in titration studies indicating significant effect of rapid temperature decrease on equilibrium of Cu(II) complexes. We propose the molecular structure of all the studied complexes at the glassy state deduced from EPR and optical spectroscopy results., (Copyright © 2017 Elsevier Inc. All rights reserved.)

Mineralization of the skeleton starts within cell-derived matrix vesicles (MVs); then, minerals propagate to the extracellular collagenous matrix. Tissue-nonspecific alkaline phosphatase (TNAP) degrades inorganic pyrophosphate (PP i ), a potent inhibitor of mineralization, and contributes P i (Phosphate) from ATP to initiate mineralization. Compared to the plasma membrane, MVs are rich in Cholesterol (Chol) (∼32%) and TNAP, but how Chol influences TNAP activity remains unclear. We have reconstituted TNAP in liposomes of dipalmitoylphosphatidylcholine (DPPC) or dioleoylphosphatidylcholine (DOPC) combined with Chol or its derivatives Cholestenone (Achol) and Ergosterol (Ergo). DPPC plus 36% sterols in liposome increased the catalytic activity of TNAP toward ATP. The presence of Chol also increased the propagation of minerals by 3.4-fold. The catalytic efficiency of TNAP toward ATP was fourfold lower in DOPC proteoliposomes as compared to DPPC proteoliposomes. DOPC proteoliposomes also increased biomineralization by 2.8-fold as compared to DPPC proteoliposomes. TNAP catalyzed the hydrolysis of ATP more efficiently in the case of the proteoliposome consisting of DOPC with 36% Chol. The same behavior emerged with Achol and Ergo. The organization of the lipid and the structure of the sterol influenced the surface tension (γ), the TNAP phosphohydrolytic activity in the monolayer, and the TNAP catalytic efficiency in the bilayers. Membranes in the Lα phase (Achol) provided better kinetic parameters as compared to membranes in the Lo phase (Chol and Ergo). In conclusion, the physical properties and the lateral organization of lipids in proteoliposomes are crucial to control mineral propagation mediated by TNAP activity during mineralization., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Tooth hypersensitivity is a growing problem affecting both the young and ageing population worldwide. Since an effective and permanent solution is not yet available, we propose a new methodology for the restoration of dental enamel using femtosecond lasers and novel calcium phosphate biomaterials. During this procedure the irradiated mineral transforms into a densified layer of acid resistant iron doped β-pyrophosphate, bonded with the surface of eroded enamel. Our aim therefore is to evaluate this densified mineral as a potential replacement material for dental hard tissue. To this end, we have tested the hardness of β-pyrophosphate pellets (sintered at 1000°C) and its mineral precursor (brushite), the wear rate during simulated tooth-brushing trials and the cytocompatibility of these minerals in powder form. It was found that the hardness of the β-pyrophosphate pellets is comparable with that of dental enamel and significantly higher than dentine while, the brushing trials prove that the wear rate of β-pyrophosphate is much slower than that of natural enamel. Finally, cytotoxicity and genotoxicity tests suggest that iron doped β-pyrophosphate is cytocompatible and therefore could be used in dental applications. Taken together and with the previously reported results on laser irradiation of these materials we conclude that iron doped β-pyrophosphate may be a promising material for restoring acid eroded and worn enamel., (Crown Copyright © 2017. Published by Elsevier B.V. All rights reserved.)

Background: Extruded rice grains are often cofortified with iron and zinc. However, it is uncertain if the addition of zinc to iron-fortified rice affects iron absorption and whether this is zinc-compound specific. Objective: We investigated whether zinc, added as zinc oxide (ZnO) or zinc sulfate (ZnSO 4 ), affects human iron absorption from extruded rice fortified with ferric pyrophosphate (FePP). Methods: In 19 iron-depleted Swiss women (plasma ferritin ≤16.5 μ/L) aged between 20 and 39 y with a normal body mass index (in kg/m 2 ; 18.7-24.8), we compared iron absorption from 4 meals containing fortified extruded rice with 4 mg Fe and 3 mg Zn. Three of the meals contained extruded rice labeled with FePP ( 57 FePP): 1 ) 1 meal without added zinc ( 57 FePP-Zn), 2 ) 1 cofortified with ZnO ( 57 FePP+ZnO), and 3 ) 1 cofortified with ZnSO 4 ( 57 FePP+ZnSO 4 ). The fourth meal contained extruded rice without iron or zinc, extrinsically labeled with ferrous sulfate ( 58 FeSO 4 ) added as a solution after cooking. All 4 meals contained citric acid. Iron bioavailability was measured by isotopic iron ratios in red blood cells. We also measured relative in vitro iron solubility from 57 FePP-Zn, 57 FePP+ZnO, and 57 FePP+ZnSO 4 expressed as a fraction of FeSO 4 solubility. Results: Geometric mean fractional iron absorption (95% CI) from 57 FePP+ZnSO 4 was 4.5% (3.4%, 5.8%) and differed from 57 FePP+ZnO (2.7%; 1.8%, 4.1%) ( P < 0.03); both did not differ from 57 FePP-Zn: 4.0% (2.8%, 5.6%). Relative iron bioavailabilities compared with 58 FeSO 4 were 62%, 57%, and 38% from 57 FePP+ZnSO 4 , 57 FePP-Zn, and 57 FePP+ZnO, respectively. In vitro solubility from 57 FePP+ZnSO 4 differed from that of 57 FePP-Zn (14.3%; P < 0.02) but not from that of 57 FePP+ZnO (10.2% compared with 13.1%; P = 0.08). Conclusions: In iron-depleted women, iron absorption from FePP-fortified extruded rice cofortified with ZnSO 4 was 1.6-fold (95% CI: 1.4-, 1.9-fold) that of rice cofortified with ZnO. These findings suggest that ZnSO 4 may be the preferable zinc cofortificant for optimal iron bioavailability of iron-fortified extruded rice. This trial was registered at clinicaltrials.gov as NCT02255942., (© 2017 American Society for Nutrition.)

Ionotropic gelation of chitosan by means of opposite charged ions represents an efficient alternative to covalent reticulation because of milder condition of use and, in general, higher biocompatibility of the resulting systems. In this work 90° light scattering (turbidimetry), circular dichroism (CD) and 1 H NMR measurements have been performed to study the interactions between the biopolymer and ionic cross-linkers tripolyphosphate (TPP) and pyrophosphate (PPi) in dilute solutions. Thereafter, a dialysis-based technique was exploited to fabricate tridimensional chitosan hydrogels based on both polyanions. Resulting matrices showed a different mechanical behavior because of their peculiar mesh-texture at micro/nano-scale: in the present contribution we demonstrate that TPP and PPi favor the formation of homogeneous and inhomogeneous systems, respectively. The different texture of networks could be exploited in future for the preparation of systems for the controlled release of molecules., (Copyright © 2016 Elsevier B.V. All rights reserved.)

An automated stopped-in-loop flow analysis (SILFA) system is proposed for the successive catalytic determination of vanadium and iron. The determination of vanadium was based on the p-anisidine oxidation by potassium bromate in the presence of Tiron as an activator to form a reddish dye, which has an absorption maximum at 510 nm. The selectivity of the vanadium determination was greatly improved by adding diphosphate as a masking agent of iron. For the iron determination, an iron-catalyzed oxidative reaction of p-anisidine by hydrogen peroxide with 1,10-phenanthroline as an activator to produce a reddish dye (510 nm) was employed. The SILFA system consisted of two peristaltic pumps, two six-port injection valves, a four-port selection valve, a heater device, a spectrophotometric detector and a data acquisition device. One six-port injection valve was used for the isolation of a mixed solution of standard/sample and reagent to promote each catalytic reaction, and another six-port injection valve was used for switching the reagent for vanadium or iron to achieve selective determination of each analyte. The above mentioned four-port selection valve was used to select standard solutions or sample. These three valves and the two peristaltic pumps were controlled by a built-in programmable logic controller in a touchscreen controller. The obtained results showed that the proposed SILFA monitoring system constituted an effective approach for the selective determination of vanadium and iron. The limits of detection, 0.052 and 0.55 µg L(-1), were obtained for vanadium and iron, respectively. The proposed system was successfully applied to drinking water samples without any preconcentration procedures., (Copyright © 2015 Elsevier B.V. All rights reserved.)

A chemiluminescent method is proposed for quantitation of NO generation in cell cultures. The method is based on activation of soluble guanylyl cyclase by NO. The product of the guanylyl cyclase reaction, pyrophosphate, is converted to ATP by ATP sulfurylase and ATP is detected in a luciferin-luciferase system. The method has been applied to the measurement of NO generated by activated murine macrophages (RAW 264.7) and bovine aortic endothelial cells. For macrophages activated by lipopolysaccharide and γ-interferon, the rate of NO production is about 100 amol/(cell·min). The rate was confirmed by the measurements of nitrite, the product of NO oxidation. For endothelial cells, the basal rate of NO generation is 5 amol/(cell·min); the rate approximately doubles upon activation by bradykinin, Ca(2+) ionophore A23187 or mechanical stress. For both types of cells the measured rate of NO generation is strongly affected by inhibitors of NO synthase. The sensitivity of the method is about 50 pM/min, allowing the registration of NO generated by 10(2)-10(4) cells. The enzyme-linked chemiluminescent method is two orders of magnitude more sensitive than fluorescent detection using 4-amino-5-methylamino-2',7'-difluorofluorescein (DAF-FM)., (Copyright © 2015 Elsevier Inc. All rights reserved.)

An anthracene-armed tetraaza macrocyclic fluorescent probe 3-(9-anthrylmethyl)-3,6,9,15-tetraazabicyclo[9.3.1]pentadeca-1(15),11,13-triene(l) for detecting Zn(2+) in aqueous medium was synthesized. L-Zn(2+) complex, showed selectivity toward pyrophosphate ion (PPi) by quenching the fluorescence in aqueous HEPES buffer (pH 7.4). Furthermore, L-Zn(2+) was also used to set up a real-time fluorescence assay for monitoring enzyme activities of alkaline phosphatase (ALP) and adenosine triphosphate sulfurylase (ATPS). In the presence of ALP inhibitor Na3VO4 and ATPS inhibitor chlorate, two enzymes activities decreased obviously, respectively., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Glucose-1-phosphate uridylyltransferase in conjunction with UDP-glucose pyrophosphorylase was found to catalyse the conversion of a range of 5-substituted UTP derivatives into the corresponding UDP-galactose derivatives in poor yield. Notably the 5-iodo derivative was not converted to UDP-sugar. In contrast, UDP-glucose pyrophosphorylase in conjunction with inorganic pyrophosphatase was particularly effective at converting 5-substituted UTP derivatives, including the iodo compound, into a range of gluco-configured 5-substituted UDP-sugar derivatives in good yields. Attempts to effect 4"-epimerization of these 5-substituted UDP-glucose with UDP-glucose 4"-epimerase from yeast were unsuccessful, while use of the corresponding enzyme from Erwinia amylovora resulted in efficient epimerization of only 5-iodo-UDP-Glc, but not the corresponding 5-aryl derivatives, to give 5-iodo-UDP-Gal. Given the established potential for Pd-mediated cross-coupling of 5-iodo-UDP-sugars, this provides convenient access to the galacto-configured 5-substituted-UDP-sugars from gluco-configured substrates and 5-iodo-UTP., (Copyright © 2015 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Objectives: Pyrophosphate (PPi) is a potent inhibitor of ectopic mineralization but its role during aortic valve calcification is not known., Methods: Anti-calcific effect of PPi was investigated by using an in vitro model of serum-driven calcification of collagen sponges and decellularized porcine aortic valve leaflets. Bovine interstitial valve cells (VIC), seeded either within the collagen matrices or in transwell chambers, were used to test cellular ability to inhibit serum-induced calcification. PPi metabolism was investigated in clonal VIC harboring different calcifying potential., Results: In a cell-free system, high serum levels induced a dose-dependent calcification of type I collagen matrices which was prevented by PPi and ATP supplementation. Blockade of serum-driven calcification by PPi and ATP was also observed when using decellularized porcine aortic valve leaflets. A similar anti-calcific effect was also seen for bovine VIC, either statically seeded into the collagen matrices or co-cultured by using a transwell system. However, when we performed co-culture experiments by using clonal VIC harboring different calcifying potential, we observed that the subset of cells acquiring a pro-calcific profile lost the ability to protect the collagen from serum-driven calcification. Pro-calcific differentiation of the clonal VIC was accompanied by increase in ALP along with significant reduction in NPP activity and ATP/PPi extracellular accumulation. These changes were not observed in the clonal subtype with lower propensity towards calcification., Conclusions: We showed that PPi and ATP are potent inhibitors of serum-driven calcification of collagen matrix and that their extracellular accumulation is reduced in calcifying VIC., (Copyright © 2014 Elsevier Ireland Ltd. All rights reserved.)

Effects of encapsulated sodium tripolyphosphate (STP), sodium hexametaphosphate (HMP) and sodium pyrophosphate (SPP) on lipid oxidation in uncooked (0, 2, 24h) and cooked (0, 1, 7 d) ground chicken and beef during storage were determined. Ten phosphate treatments included a control (no phosphate), three unencapsulated (u) at 0.5% and three encapsulated (e) phosphates (0.5%) each at a low (e-low) and high (e-high) coating level. Two heating rates (slow, fast) were investigated. Cooking loss (CL), pH, color, orthophosphate (OP), TBARS and lipid hydroperoxides (LPO) were determined. A fast heating and uSTP resulted in lower CL (p<0.05). Orthophosphate increased with phosphate incorporation, slow heating and storage (p<0.05). Encapsulated phosphates and increased coating level reduced OP (p<0.05). Unencapsulated STP increased CIE a* and pH, whereas uSPP decreased CIE a* and pH (p<0.05). Encapsulated phosphates and the greater coating level had no effect on the pH in cooked samples. Not increased coating level but encapsulated phosphates decreased lipid oxidation in cooked samples (p<0.05)., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Ionically crosslinked chitosan particles with submicron dimensions attract widespread interest as materials for controlled release. To this end, we have examined the formation and dissolution of nanoparticles prepared by crosslinking chitosan with pyrophosphate (PPi). The formation of these particles required a critical PPi concentration (which increased with the chitosan concentration), and their z-average hydrodynamic diameters could be predictably tuned from roughly 60 to 220 nm by varying the concentration of the parent chitosan solutions. Unlike the nanoparticles crosslinked with the commonly used tripolyphosphate (TPP), which coagulated and precipitated when TPP was in excess, the chitosan/PPi nanoparticles remained colloidally stable even at high PPi concentrations. Moreover, the analysis of their dissolution revealed hysteresis in the particle formation/dissolution cycle, where portions of the crosslinked chitosan/PPi complexes remained stably intact at PPi concentrations below those required for their formation. This irreversible behavior was surmised to reflect the cooperativity of chitosan/PPi binding and was qualitatively modeled using the Bragg-Williams theory., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Due to a fast setting reaction, good biological properties, and easily available starting materials, there has been extensive research within the field of brushite cements as bone replacing material. However, the fast setting of brushite cement gives them intrinsically low mechanical properties due to the poor crystal compaction during setting. To improve this, many additives such as citric acid, pyrophosphates, and glycolic acid have been added to the cement paste to retard the crystal growth. Furthermore, the incorporation of a filler material could improve the mechanical properties when used in the correct amounts. In this study, the effect of the addition of the two retardants, disodium dihydrogen pyrophosphate and citric acid, together with the addition of β-TCP filler particles, on the mechanical properties of a brushite cement was investigated. The results showed that the addition of low amounts of a filler (up to 10%) can have large effects on the mechanical properties. Furthermore, the addition of citric acid to the liquid phase makes it possible to use lower liquid-to-powder ratios (L/P), which strongly affects the strength of the cements. The maximal compressive strength (41.8MPa) was found for a composition with a molar ratio of 45:55 between monocalcium phosphate monohydrate and beta-tricalcium phosphate, an L/P of 0.25ml/g and a citric acid concentration of 0.5M in the liquid phase., (Copyright © 2013 The Authors. Published by Elsevier Ltd.. All rights reserved.)

The analysis of free amino acids in urine and plasma is useful for estimating disease status in clinical diagnoses. Changes in the concentration of free amino acids in foods are also useful markers of freshness, nutrition, and taste. In this study, the specific interaction between aminoacyl-tRNA synthetase (aaRS) and its corresponding amino acid was used to measure amino acid concentrations. Pyrophosphate released by the amino acid-aaRS binding reaction was detected by luminol chemiluminescence; the method provided selective quantitation of 1.0-30 μM histidine and 1.0-60 μM lysine., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Ap4A hydrolase (asymmetrical diadenosine tetraphosphate hydrolase, EC 3.6.1.17), an enzyme involved in a number of biological processes, is characterized as cleaving the polyphosphate chain at the fourth phosphate from the bound adenosine moiety. This paper presents the crystal structure of wild-type and E58A mutant human Ap4A hydrolase. Similar to the canonical Nudix fold, human Ap4A hydrolase shows the common αβα-sandwich architecture. Interestingly, two sulfate ions and one diphosphate coordinated with some conserved residues were observed in the active cleft, which affords a better understanding of a possible mode of substrate binding., (Copyright © 2013 Elsevier Inc. All rights reserved.)

The diphosphoinositol polyphosphates ("inositol pyrophosphates"; PP-InsPs) regulate many cellular processes in eukaryotes, including stress responses, apoptosis, vesicle trafficking, cytoskeletal dynamics, exocytosis, telomere maintenance, insulin signaling and neutrophil activation. Thus, the enzymes that control the metabolism of the PP-InsPs serve important cell signaling roles. In order to fully characterize how these enzymes are regulated, we need to determine the atomic-level architecture of their active sites. Only then can we fully appreciate reaction mechanisms and their modes of regulation. In this review, we summarize published information obtained from the structural analysis of a human diphosphoinositol polyphosphate phosphohydrolase (DIPP), and a human diphosphoinositol polyphosphate kinase (PPIP5K). This work includes the analysis of crystal complexes with substrates, products, transition state analogs, and a novel phosphonoacetate substrate analog., (Published by Elsevier Ltd.)

We have investigated the applicability of different chemical methods for pyrophosphate bond formation to the synthesis of 5-substituted UDP-galactose and UDP-N-acetylglucosamine derivatives. The use of phosphoromorpholidate chemistry, in conjunction with N-methyl imidazolium chloride as the promoter, was identified as the most reliable synthetic protocol for the preparation of these non-natural sugar-nucleotides. Under these conditions, the primary synthetic targets 5-iodo UDP-galactose and 5-iodo UDP-N-acetylglucosamine were consistently obtained in isolated yields of 40-43%. Both 5-iodo UDP-sugars were used successfully as substrates in the Suzuki-Miyaura cross-coupling with 5-formylthien-2-ylboronic acid under aqueous conditions. Importantly, 5-iodo UDP-GlcNAc and 5-(5-formylthien-2-yl) UDP-GlcNAc showed moderate inhibitory activity against the GlcNAc transferase GnT-V, providing the first examples for the inhibition of a GlcNAc transferase by a base-modified donor analogue., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Objective: (99m)Tc-Sn-PYP (Technetium-99(m) labeled tin pyrophosphate) has been widely used as a radiopharmaceutical for bone scanning as well as in nuclear cardiology. It is also found in the body in trace amounts. (177)Lu is presently considered as an excellent radionuclide for developing bone pain palliation agents. PYP is an analogue of MDP and MDP has been labeled with (177)Lu. No study on preparing a complex of (177)Lu with PYP has been reported yet. Based on these facts, it was hypothesized that a bone-seeking (177)Lu-PYP (Lutetium-177 labeled Pyrophosphate) radiopharmaceutical could be developed as an agent for palliative radiotherapy of bone pain due to skeletal metastases., Methods: (177)Lu was produced by irradiating lutetium foil (11 mg) natural target at a flux ∼1.0×10(14)n/cm(2)/s for 12 h in the swimming pool type reactor. (177)Lu in the form of (177)LuCl(3) was labeled with PYP. The radiochemical purity and labeling efficiencies were determined by paper chromatography. Labeling of (177)Lu with PYP was optimized and a labeled sample was subjected to HPLC analysis. To determine the charge on the (177)Lu-PYP complex, radio-electrophoresis was conducted for 1 h under a voltage of 300 V and 45 mA current using 0.025 M phosphate buffer (pH 6.9). Bioevaluation studies with rabbit under γ-camera were also performed to verify the skeletal uptake., Results: The quality control using paper radio-chromatography has shown >99% radiochemical purity of (177)Lu-PYP complex. Radio-chromatography also showed maximum labeling at ligand/metal ratio=60:1. HPLC analysis showed 1.42±0.01 min retention time of (177)Lu-PYP complex. No decrease in labeling was observed at higher temperatures. Gamma-camera images of (177)Lu-PYP in normal rabbit at 24 h post injection also showed high skeletal uptake., Conclusion: The study demonstrated that sodium pyrophosphate could be labeled with (177)Lu with high radiochemical yields (>99%). Negatively charged (177)Lu-PYP complex retained stability for a day and at high temperatures too. Gamma-camera images of (177)Lu-PYP in normal rabbit at 24 h post injection showed high skeletal uptake, suggesting that it may be useful as a bone-pain palliation agent for the treatment of bone metastases., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Validated methods for the determination of CA1, the active agent derived from the prodrug CA1P, in human plasma and urine, and of CA1P and three glucuronides CA1G1, CA1G2 and CA1DG in human urine were developed using LC-MS. Plasma CA1 was extracted using solid phase extraction and validated over the range 5-1000 nM. Urine samples were analysed without extraction, and the assays validated over the range 50-2000 nM (CA1P), 25-2000 nM (CA1), 50-40,000 nM (CA1G1 and CA1G2) and 25-4000 nM (CA1DG). The mean correlation coefficient (r²) was ≥ 0.997 for all assays. The intra-day and inter-day accuracy and precision were within the generally accepted criteria for bioanalytical methods (<15%). Mean recovery of CA1 from plasma was 101%, and 97% from urine. Mean urine recovery of CA1P was 98%, CA1G1 96%, CA1G2 93% and CA1DG 93%. The method was applied to plasma and urine samples from a recently completed clinical trial of the prodrug. Peak plasma concentrations of up to 470 nM CA1 were seen. The majority of drug-related material measured in urine comprised of the two monoglucuronides; CA1 and the diglucuronide were about 10-fold lower. No CA1P was detectable in urine., (Copyright © 2012 Elsevier B.V. All rights reserved.)

This work focuses on the remediation of polycyclic aromatic hydrocarbon (PAH)-contaminated soil using modified Fenton (MF) treatment coupled with a novel chelating agent (CA), a more effective technique among currently available technologies. The performance of MF treatment to promote PAH oxidation in artificially contaminated soil was investigated in a packed column with a hydrogen peroxide (H(2)O(2)) delivery system simulating in-situ soil flushing which is more representative of field conditions. The effectiveness of process parameters H(2)O(2)/soil, Fe(3+)/soil, CA/soil weight ratios and reaction time were studied using a 2(4) three level factorial design experiments. An optimised operating condition of the MF treatment was observed at H(2)O(2)/soil 0.05, Fe(3+)/soil 0.025, CA/soil 0.04 and 3h reaction time with 79.42% and 68.08% PAH removals attainable for the upper and lower parts of the soil column respectively. The effects of natural attenuation and biostimulation process as post-treatment in the remediation of the PAH-contaminated soil were also studied. In all cases, 3-aromatic ring PAH (phenanthrene) was more readily degraded than 4-aromatic ring PAH (fluoranthene) regardless of the bioremediation approach. The results revealed that both natural attenuation and biostimulation could offer remarkable enhancement of up to 6.34% and 9.38% in PAH removals respectively after 8 weeks of incubation period. Overall, the results demonstrated that combined inorganic CA-enhanced MF treatment and bioremediation serves as a suitable strategy to enhance soil quality particularly to remediate soils heavily contaminated with mixtures of PAHs., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Terpenoids form the largest class of plant metabolites involved in primary and secondary metabolism. Isoprenyl diphosphate synthases (IDSs) catalyze the condensation of the C(5) terpenoid building blocks, isopentenyl diphosphate and dimethylallyl diphosphate, to form geranyl diphosphate (C(10)), farnesyl diphosphate (C(15)), and geranylgeranyl diphosphate (C(20)). These branch point reactions control the flow of metabolites that act as precursors to each of the major terpene classes-monoterpenes, sequiterpenes, and diterpenes, respectively. Thus accurate and easily performed assays of IDS enzyme activity are critical to increase our knowledge about the regulation of terpene biosynthesis. Here we describe a new and sensitive nonradioactive method for carrying out IDS assays using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) to detect the short-chain prenyl diphosphate products directly without dephosphorylation. Furthermore, we were able to separate cisoid and transoid isomers of both C(10) enzyme products (geranyl diphosphate and neryl diphosphate) and three C(15) products [(E,E)-, (Z,E)-, and (Z,Z)-farnesyl diphosphate]. By applying the method to crude protein extracts from various organs of Arabidopsis thaliana, Nicotiana attenuata, Populus trichocarpa, and Picea abies, we could determine their IDS activity in a reproducible fashion., (Copyright © 2011 Elsevier Inc. All rights reserved.)

This study investigated the influence of flushing duration, [S,S]-ethylenediaminedisuccinic acid (EDDS) dosage, humic acid and various combinations of ethylenediaminetetraacetic acid (EDTA), EDDS and tetrasodium pyrophosphate (Na(4)P(2)O(7)) on metal extraction during soil flushing, through column experiments. A lesser extent of enhancement in metal extraction efficiencies was found when the flushing duration and the dosage of EDDS was doubled, compared to their efficiencies measured at pore volume 100. Metal extraction efficiency was mainly influenced by the initial metal distribution in the soils rather than the flushing duration and the EDDS-to-metal molar ratio. Humic acid of less than 10mg/L as dissolved organic carbon (DOC) posed an insignificant effect on metal extraction during EDDS enhanced soil flushing. The extraction rate of Ni by EDTA and EDDS was time dependent, and was initially fast in the case of EDDS, whereas it was slow for EDTA. However, the overall Ni extraction efficiency by EDTA was higher when the flushing time was longer. Na(4)P(2)O(7) promoted the mineral dissolution which enhanced the metal extraction as a result of soil disruption. The order of metal extraction by Na(4)P(2)O(7) was Ni>Cr>Cu, probably be due to the different affinities between metals and P(2)O(7)(4-)., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The optimization of analytical procedures for the quantification of free and total microcystins (MCs) in natural sediments was systematically examined based on solvent extraction and Lemieux oxidation. In this optimized analytical procedure, a sequential solvent extraction using 50% (v/v) methanol and EDTA-sodium pyrophosphate was selected as the optimal extraction solvent for free MCs analysis, after which the purified extracts and sediment residuals were applied to the optimized Lemieux oxidation for determination of total MCs in lake sediments. The optimized procedures were shown to be efficient and reliable for the routine analysis of both free and total MCs in lake sediment samples, as indicated by the minimal adverse impact of sediment organic matter on the recovery of free MCs and yield of MMPB (2-methyl-3-methoxy-4-phenylbutyric acid). Finally, the developed procedures were applied to field sediment samples collected from Lake Dianchi during a bloom season and seven of thirty samples showed positive results., (Copyright © 2011 Elsevier B.V. All rights reserved.)

The acid-catalysed, aqueous phase dehydration of xylose (a monosaccharide obtainable from hemicelluloses, e.g., xylan) to furfural was investigated using vanadium phosphates (VPO) as catalysts: the precursors, VOPO(4)·2H(2)O, VOHPO(4)·0.5H(2)O and VO(H(2)PO(4))(2), and the materials prepared by calcination of these precursors, that is, γ-VOPO(4), (VO)(2)P(2)O(7) and VO(PO(3))(2), respectively. The VPO precursors were completely soluble in the reaction medium. In contrast, the orthorhombic vanadyl pyrophosphate (VO)(2)P(2)O(7), prepared by calcination of VOHPO(4)·0.5H(2)O at 550°C/2 h, could be recycled by simply separating the solid acid from the reaction mixture by centrifugation, and no drop in catalytic activity and furfural yields was observed in consecutive 4 h-batch runs (ca. 53% furfural yield, at 170°C). However, detailed catalytic/characterisation studies revealed that the vanadyl pyrophosphate acts as a source of active water-soluble species in this reaction. For a concentration of (VO)(2)P(2)O(7) as low as 5 mM, the catalytic reaction of xylose (ca. 0.67 M xylose in water, and toluene as solvent for the in situ extraction of furfural) gave ca. 56% furfural yield, at 170°C/6 h reaction., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

Carbon nanotubes (CNTs) can efficiently quench the fluorescence of the adsorbed fluorophores and nonconvalently interact with soft single-stranded DNA (ssDNA). Upon disruption of CNTs-fluorescent oligonucleotides hybrid by nuclease S1, fluorescence turn-on was observed. Using this strategy, a platform based on fluorescence signal for monitoring the activity of nuclease with advantages of high sensitivity and commonality was established, and a linear relationship between initial cleavage reaction rate and nuclease S1 concentration is found in the range of 0.6-8.0 U mL(-1) with a detection limit of 0.08 U mL(-1). Furthermore, by taking pyrophosphate as an example, we use the assay to evaluate the prohibition effect on nuclease, and the extent of fluorescence recovery decreased linearly with increasing the concentration of pyrophosphate in the range of 0.2-1.4 mM, implying that the cleavage reaction by nuclease S1 was prohibited, and therefore this fluorescence assay can also be conveniently utilized for inhibitor screening of nuclease., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Bacteria monitoring is essential for many industrial manufacturing processes, particularly those involving in food, biopharmaceuticals, and semiconductor production. Firefly luciferase ATP luminescence assay is a rapid and simple bacteria detection method. However, the detection limit of this assay for Escherichia coli is approximately 10(4) colony-forming units (CFU), which is insufficient for many applications. This study aims to improve the assay sensitivity by simultaneous conversion of PP(i) and AMP, two products of the luciferase reaction, back to ATP to form two chain-reaction loops. Because each consumed ATP continuously produces two new ATP molecules, this approach can achieve exponential amplification of ATP. Two consecutive enzyme reactions were employed to regenerate AMP into ATP: adenylate kinase converting AMP into ADP using UTP as the energy source, and acetate kinase catalyzing acetyl phosphate and ADP into ATP. The PP(i)-recycling loop was completed using ATP sulfurylase and adenosine 5' phosphosulfate. The modification maintains good quantification linearity in the ATP luminescence assay and greatly increases its bacteria detection sensitivity. This improved method can detect bacteria concentrations of fewer than 10 CFU. This exponential ATP amplification assay will benefit bacteria monitoring in public health and manufacturing processes that require high-quality water., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Double-rotation (DOR) is the only technique generally capable of yielding high-resolution NMR spectra of half-integer quadrupolar nuclei in one dimension for solids without the need for sophisticated coherence pathway selection. Unfortunately, due to the low outer rotor spinning frequencies currently available, the spectra often contain a large number of spinning sidebands which may overlap with the resonances of interest. We implement a simple, robust, and easy to use family of pulse sequences, which in practice are fully analogous to the 'total suppression of sidebands' (TOSS) sequences, to suppress all sidebands arising from the spinning of the outer rotor in DOR experiments. By removing the rotor phase dependence of the evolution of the sidebands, the sidebands destructively interfere with one another during the course of signal averaging to yield 'solution-like' spectra of half-integer quadrupolar nuclei in solids. Advantages and shortcomings of the method compared to other DOR sideband suppression methods are explored with the aid of simulations., (Copyright © 2011 Elsevier Inc. All rights reserved.)

Phosphate buffer is frequently used in biological, biochemical and biomedical applications especially when pH is to be controlled around the physiological value of 7.4. One of the prerequisites of a buffer compound among good buffering capacity and pH stability over time is its non-reactivity with other constituents of the solution. This is especially important for quantitative analytical or toxicological assays. Previous work has identified a number of amino alcohol buffers like TRIS to react with G-type nerve agents sarin, soman and cyclosarin to form stable phosphonic diesters. In case of phosphate buffer we were able to confirm not only the rapid hydrolysis of these agents to the respective alkyl methylphosphonates but also the formation of substantial amounts of pyrophosphate-like adducts (phosphorylated methylphosphonates), which very slowly hydrolyzed following zero-order kinetics. This led to a complex mixture of phosphorus containing species with changing concentrations over time. We identified the molecular structure of these buffer adducts using 1D ¹H-³¹P HSQC NMR and LC-ESI-MS/MS techniques. Reaction rates of adduct formation are fast enough to compete with hydrolysis in aqueous solution and to yield substantial amounts of buffer adduct over the course of just a couple of minutes. Possible reaction mechanisms are discussed with respect to the formation and subsequent hydrolysis of the pyrophosphate-like compounds as well as the increased rate of hydrolysis of the nerve agent to the corresponding alkyl methylphosphonates. In summary, the use of phosphate buffer for the development of new assays with sarin, soman and cyclosarin is discouraged. Already existing protocols should be carefully reexamined on an individual basis., (Copyright © 2010 Elsevier Ireland Ltd. All rights reserved.)

The kinetics of the oxidative degradation of dipeptide glycyl-glycine (Gly-Gly) by water-soluble colloidal MnO(2) in acidic medium has been studied by employing visible spectrophotometer in the aqueous and micellar media at 35°C. To obtain the rate constants as functions of [Gly-Gly], [MnO(2)] and [HClO(4)], pseudo-first-order conditions were maintained in each kinetic run. The first-order-rate is observed with respect to [MnO(2)], whereas fractional-order-rates are determined in both [Gly-Gly] and [HClO(4)]. The addition of sodium pyrophosphate and sodium fluoride has composite effects (catalytic and inhibition). The reaction proceeds through the fast adsorption of Gly-Gly on the surface of the colloidal MnO(2). The observed results are discussed in terms of Michaelis-Menten/Langmuir-Hinshelwood model. The Arrhenius and Eyring equations are found valid for the reaction over a range of temperatures and different activation parameters have been evaluated. A probable reaction mechanism, in agreement with the observed kinetic results, has been proposed and discussed. The influence of changes in the surfactant concentrations on the observed rate constant is also investigated and the reaction followed the same type of kinetic behavior in micellar media. The pseudo-first-order rate constant (k(ψ)) is found to increase about two-fold with increase in [TX-100]. The catalytic effect of nonionic surfactant TX-100 is explained in terms of the mathematical model proposed by Tuncay et al., (Copyright © 2010 Elsevier B.V. All rights reserved.)