A photoluminescent and electrochemiluminescent probe based on an iridium(III) complex with a boronic acid-functionalised ancillary ligand for the selective detection of mercury(II) ions.

Exposure to mercury(II) ions (Hg2+) can cause various diseases such as Minamata disease, acrodynia, Alzheimer's disease, and Hunter-Russell syndrome, and even organ damage. Therefore, real-time and accurate monitoring of Hg2+ in environmental samples is crucial. In this study, we report a photoluminescent (PL) and electrochemiluminescent (ECL) probe based on a cyclometalated Ir(III) complex for the selective detection of Hg2+. The introduction of a reaction site, o-aminomethylphenylboronic acid, on the ancillary ligands allowed a prompt transmetalation reaction to take place between Hg2+ and boronic acid. This reaction resulted in significant decreases of the PL and ECL signals due to the photo-induced electron transfer from the Ir(III) complex to the Hg2+ ions. The probe was applied to the selective detection of Hg2+, and the signal changes revealed a linear correlation with Hg2+ concentrations in the range of 0-10 µM (LOD = 0.72 µM for PL, 8.03 nM for ECL). The designed probe allowed the successful quantification of Hg2+ in tap water samples, which proves its potential for the selective detection of Hg2+ in environmental samples.

Highly Selective Electrochemiluminescence Chemosensor for Sulfide Enabled by Hierarchical Reactivity.

Hydrogen sulfide (H2S) is a well-known toxic gas with the odor of rotten eggs. Several reaction-based electrochemiluminescence (ECL) chemosensors for H2S have been developed; however, no homogeneous ECL probe with high selectivity toward H2S in aqueous media has been reported. Herein, we report an iridium(III) complex-based ECL chemodosimetric probe employing two 7-nitrobenz-2-oxa-1,3-diazol-4-yl (NBD) groups known as a photo-induced electron transfer quencher and a reaction site for the selective detection of H2S; the detection mechanism involves H2S being clearly distinguished from biothiols based on the different cleavage rates of the two NBD groups and extremely weak ECL interferences caused by reaction by-products. The probe was rationally designed to improve selectivity toward H2S within the ECL analysis platform by enabling the removal of nonspecific background signals observed via fluorescence analysis. This analytical system exhibited remarkable selectivity toward H2S, a rapid reaction rate, and high sensitivity (LOD = 57 nM) compared to conventional fluorescence methods. Furthermore, the probe could successfully quantify H2S in tap water samples and commercial ammonium sulfide solutions, which demonstrates the effectiveness of this probe in field monitoring.

Ferroelectric PVDF nanofiber membrane for high-efficiency PM0.3 air filtration with low air flow resistance.

The significant public health concerns related to particulate matter (PM) air pollutants and the airborne transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have led to considerable interest in high-performance air filtration membranes. Highly ferroelectric polyvinylidene fluoride (PVDF) nanofiber (NF) filter membranes are successfully fabricated via electrospinning for high-performance low-cost air filtration. Spectroscopic and ferro-/piezoelectric analyses of PVDF NF show that a thinner PVDF NF typically forms a ferroelectric ß phase with a confinement effect. A 70-nm PVDF NF membrane exhibits the highest fraction of ß phase (87%) and the largest polarization behavior from piezoresponse force microscopy. An ultrathin 70-nm PVDF NF membrane exhibits a high PM0.3 filtration efficiency of 97.40% with a low pressure drop of 51 Pa at an air flow of 5.3 cm/s owing to the synergetic combination of the slip effect and ferroelectric dipole interaction. Additionally, the 70-nm PVDF NF membrane shows excellent thermal and chemical stabilities with negligible filtration performance degradation (air filtration efficiency of 95.99% and 87.90% and pressure drop of 55 and 65 Pa, respectively) after 24 h of heating at 120 °C and 1 h immersion in isopropanol.

The Stringent Response Contributes to Persistent Methicillin-Resistant Staphylococcus aureus Endovascular Infection Through the Purine Biosynthetic Pathway.

Persistent methicillin-resistant Staphylococcus aureus (MRSA) endovascular infections represent a significant clinical-therapeutic challenge. Of particular concern is antibiotic treatment failure in infections caused by MRSA that are "susceptible" to antibiotic in vitro. In the current study, we investigate specific purine biosynthetic pathways and stringent response mechanism(s) related to this life-threatening syndrome using genetic matched persistent and resolving MRSA clinical bacteremia isolates (PB and RB, respectively), and isogenic MRSA strain sets. We demonstrate that PB isolates (vs RB isolates) have significantly higher (p)ppGpp production, phenol-soluble-modulin expression, polymorphonuclear leukocyte lysis and survival, fibronectin/endothelial cell (EC) adherence, and EC damage. Importantly, an isogenic strain set, including JE2 parental, relP-mutant and relP-complemented strains, translated the above findings into significant outcome differences in an experimental endocarditis model. These observations indicate a significant regulation of purine biosynthesis on stringent response, and suggest the existence of a previously unknown adaptive genetic mechanism in persistent MRSA infection.

Potential-Dependent Electrochemiluminescence for Selective Molecular Sensing of Cyanide.

Although tremendous efforts have been devoted to providing specificity for molecular sensors, most of the methods focus on the structural variation of the binding or reaction site to improve selectivity. Herein, we report a new approach in which a chemical probe, possessing a mediocre recognition site, can successfully discriminate a target among various interferences only with electrochemical manipulation. The synthetic probe (1) was designed to react with a cyanide anion (CN-), and its dicyanovinyl group has selectivity toward CN- along with sulfides and biothiols resulting in similar adducts. However, the binding adduct between 1 and CN- (1-CN-) has significantly different energy levels that are only able to undergo electrochemical oxidation under ∼1.2 V (vs Ag/AgCl), generating strong electrochemiluminescence (ECL). The ECL emission from 1-CN- successfully discriminates CN- without any interferences from other analytes including sulfides and thiols and exhibits a linear correlation with CN- in a range of 1-400 µM (LOD = 0.04 µM, n = 5). Density functional theory (DFT) calculations and electrochemical studies supported the mechanism of CN- discrimination. The approach was finally applied to direct trace analysis of CN- in tap water (≥1 µM) and showed excellent performance suggesting a new, versatile, and rapid determination method for molecular toxins in real samples.

Second-Harmonic Generation and Photoluminescence Properties of Sn(II)- and Bi(III)-Based Lone Pair Cation-Pyridine Dicarboxylate Coordination Compounds.

Lone pair cation-based novel coordination compounds Sn[(pdc)(H2O)] (Sn-I) and (H2bpy)[Bi(pdc)2(Hpdc)]·5H2O (Bi-I) (pdc = pyridine-2,6-dicarboxylate; bpy = 4,4'-bipyridine) were synthesized through mild hydrothermal reactions. While Sn-I crystallizing in the polar space group, Pca21, exhibits a helical chain structure consisting of SnO3N distorted seesaws, 2,6-pdc linkers, and water molecules, Bi-I crystallizing in the centrosymmetric (CS) space group, P1̅, reveals a pseudo-3D network composed of BiO5N3 polyhedra, 2,6-pdc ligands, H2bpy2+ cations, and isolated H2O molecules. The lone cations Sn2+ and Bi3+ in the title compounds are in a highly deformed polyhedral environment. The single-crystal-to-single-crystal transformation from Sn-I to the anhydrous Sn[(pdc)] (Sn-II) with the polar noncentrosymmetric structure was successfully achieved upon heating crystals of Sn-I. UV-vis diffuse reflectance spectra indicate that the introduction of Sn2+ or Bi3+ red-shifts the adsorption edges upon coordination. Powder second-harmonic generation (SHG) measurements indicate that Sn-I and Sn-II are type-I phase-matchable and exhibit SHG intensity of ca. 15 and 35 times that of α-SiO2, respectively. Solid state photoluminescence (PL) measurements indicate that Bi-I is an excellent green emitting phosphor with the quantum efficiency up to 26% and outstanding decay lifetime of 1.82 ms at room temperature.

Electrogenerated Chemiluminescent Chemodosimeter Based on a Cyclometalated Iridium(III) Complex for Sensitive Detection of Thiophenol.

Thiophenol is the simplest aromatic thiol that is utilized for various applications in industry and agriculture. However, it should be used with care because thiophenol is readily absorbed into the human body by inhalation and ingestion, which leads to serious internal injuries. Thus, there is an urgent need for real-time and accurate monitoring of thiophenol. Despite remarkable advantages of electrogenerated chemiluminescence (ECL) analysis, ECL thiophenol probes have never been reported. Herein, a new strategy for the rapid detection of thiophenol by use of an ECL turn-on chemodosimeter based on a cyclometalated Ir(III) complex is described. This analytical system showed superior sensitivity [limit of detection (LOD) value, 3.8 nM] in comparison to the conventional fluorescence method. In addition, our system exhibited remarkable selectivity and reaction rate toward thiophenol over other analytes. Moreover, it was successfully applied to quantify thiophenol in real water samples, providing a new proof-of-concept for field monitoring based on ECL.

Molecular Recognition of Pyrophosphate with Extended Bis(Zn(II)-DPA) Derivatives.

A series of pyrophosphate (PPi) receptors were synthesized, and their binding affinities toward both PPi and adenosine triphosphate were evaluated in N-(2-hydroxyethyl)piperazine-N'-ethanesulfonic acid-buffered solution. The presence of two hydrogen bond donors slightly retarded the reaction rate of anionic guest exchange, while four had a significant retardation effect. A macrocyclic receptor, 17, exhibited superior selectivity toward PPi compared with acyclic receptors, presumably due to effective steric interactions. Isothermal titration calorimetry experiments and potentiometric titration experiments revealed the effect of the substituent structure on the degree of anion guest stabilization and the deprotonation of water molecules bound to zinc(II) ions, as well as the guest binding thermodynamics.

High Throughput Bar-Coating Processed Organic-Inorganic Hybrid Multi-Layers for Gas Barrier Thin-Films.

Herein, we demonstrate the preparation of a scalable bar-coated nanocomposite organic-inorganic hybrid film and developed robust barrier films for general purpose packaging. Using combinatory printing of polymers and nanocomposites by bar coating, a facile and effective barrier film fabrication method was developed. Based on a preliminary survey with several material combinations, a rationalized two-fold nanocomposite film was fabricated. The number of layers in the barrier film significantly modified oxygen barrier performance such that, for the 1 wt% ethylene vinyl alcohol (EVOH) intercalated film, the oxygen transmission rate (OTR) of the 5-layer sample was reduced to 31.69% of the OTR of the 3-layer sample (112.8 vs. 35.75 cc/(m² · day)). In addition, fine tuning the amount of EVOH polymer enabled further improvement of oxygen barrier performance. Intercalation of 2 wt% EVOH resulted an OTR improvement from 35.75 in the 1 wt% sample to 11.90 cc/(m² ·day), which is a 4.25-fold enhancement. Overall barrier characteristics proved that our approach could be used for large-area deposited, oxygen resistant, general purpose packaging applications.

Electrostatic Modification for Promotion of Flavin-Mediated Oxidation of a Probe for Flavin Detection.

Electrostatic effects on the redox photochemistry of synthetic probes (1, 2, and 1-Zn) are examined by adjusting the thermodynamic driving force of their oxidation reactions. The redox photochemistry was simply controlled by introducing a zinc binding site (2,2'-dipicolylamine (DPA)) on the coumarin moiety of probe 2. Zinc complexation produced a positively charged environment on the coumarin (1-Zn), which lowered the electron density of a nearby 9 H-xanthene ring, attenuating the auto-oxidation of 1-Zn by 45 % compared with that of probe 1 at 298 K. The positive net charge of 1-Zn also provided an attractive Coulombic force toward the phosphate of flavin mononucleotide and flavin adenine dinucleotide, which lowered the reduction potential of the electron acceptor (isoalloxazine) and improved intermolecular electron transfer from the 9 H-xanthene ring to isoalloxazine. The flavin-mediated oxidation rate of 1-Zn was increased to 1.5 times that of probe 2. Probe 1-Zn showed highly selective sensing behaviour toward flavins, producing an intense brightness (ϵΦF =2.80×103 m-1  cm-1 ) in the long-wavelength regions (λmax =588 nm) upon flavin-mediated oxidation. Furthermore, probes 1-Zn and 2 were successfully applied to eosinophil imaging and the differential diagnosis of eosinophilia; this demonstrates their use as diagnostic tools.

Sulfatase activity assay using an activity-based probe by generation of N-methyl isoindole under reducing conditions.

Sulfatases catalyze the hydrolysis of sulfate esters that are present in a range of biomolecules. This is an important step in several biological processes such as cellular degradation, hormone regulation, and cell signaling. We have developed a new activity-based sulfatase probe (probe 1) that generates a fluorescent N-methylisoindole upon hydrolysis by sulfatase. Because of the autoxidation of N-methylisoindole, the sulfatase activity was also tested under reducing conditions, containing either glutathione (GSH) or tris(2-carboxyethyl)phosphine (TCEP), exhibiting little change in kinetic parameters compared to non-reducing conditions. Probe 1 displayed reasonable kinetic parameters under both non-reducing and reducing conditions, among which the use of Tris buffer and Tris buffer containing GSH appeared to be appropriate conditions for inhibitor screening. Probe 1 showed stronger intensity upon treatment with sulfatase under neutral conditions than under acidic conditions, but it still has limitations in the selectivity for a specific sulfatase. Nevertheless, the fluorescent signal generated as a result of the release of N-methylisoindole after treatment of probe 1 with sulfatase provides a new assay for measuring sulfatase activity that could be adapted for high throughput screening.

The bacterial alarmone (p)ppGpp activates the type III secretion system in Erwinia amylovora.

UNLABELLED: The hypersensitive response and pathogenicity (hrp) type III secretion system (T3SS) is a key pathogenicity factor in Erwinia amylovora. Previous studies have demonstrated that the T3SS in E. amylovora is transcriptionally regulated by a sigma factor cascade. In this study, the role of the bacterial alarmone ppGpp in activating the T3SS and virulence of E. amylovora was investigated using ppGpp mutants generated by Red recombinase cloning. The virulence of a ppGpp-deficient mutant (ppGpp(0)) as well as a dksA mutant of E. amylovora was completely impaired, and bacterial growth was significantly reduced, suggesting that ppGpp is required for full virulence of E. amylovora. Expression of T3SS genes was greatly downregulated in the ppGpp(0) and dksA mutants. Western blotting showed that accumulations of the HrpA protein in the ppGpp(0) and dksA mutants were about 10 and 4%, respectively, of that in the wild-type strain. Furthermore, higher levels of ppGpp resulted in a reduced cell size of E. amylovora. Moreover, serine hydroxamate and α-methylglucoside, which induce amino acid and carbon starvation, respectively, activated hrpA and hrpL promoter activities in hrp-inducing minimal medium. These results demonstrated that ppGpp and DksA play central roles in E. amylovora virulence and indicated that E. amylovora utilizes ppGpp as an internal messenger to sense environmental/nutritional stimuli for regulation of the T3SS and virulence. IMPORTANCE: The type III secretion system (T3SS) is a key pathogenicity factor in Gram-negative bacteria. Fully elucidating how the T3SS is activated is crucial for comprehensively understanding the function of the T3SS, bacterial pathogenesis, and survival under stress conditions. In this study, we present the first evidence that the bacterial alarmone ppGpp-mediated stringent response activates the T3SS through a sigma factor cascade, indicating that ppGpp acts as an internal messenger to sense environmental/nutritional stimuli for the regulation of the T3SS and virulence in plant-pathogenic bacteria. Furthermore, the recovery of an spoT null mutant, which displayed very unique phenotypes, suggested that small proteins containing a single ppGpp hydrolase domain are functional.

A regulatory feedback loop between RpoS and SpoT supports the survival of Legionella pneumophila in water.

Legionella pneumophila is a waterborne pathogen, and survival in the aquatic environment is central to its transmission to humans. Therefore, identifying genes required for its survival in water could help prevent Legionnaires' disease outbreaks. In the present study, we investigate the role of the sigma factor RpoS in promoting survival in water, where L. pneumophila experiences severe nutrient deprivation. The rpoS mutant showed a strong survival defect compared to the wild-type strain in defined water medium. The transcriptome of the rpoS mutant during exposure to water revealed that RpoS represses genes associated with replication, translation, and transcription, suggesting that the mutant fails to shut down major metabolic programs. In addition, the rpoS mutant is transcriptionally more active than the wild-type strain after water exposure. This could be explained by a misregulation of the stringent response in the rpoS mutant. Indeed, the rpoS mutant shows an increased expression of spoT and a corresponding decrease in the level of (p)ppGpp, which is due to the presence of a negative feedback loop between RpoS and SpoT. Therefore, the lack of RpoS causes an aberrant regulation of the stringent response, which prevents the induction of a successful response to starvation.

Homogeneous electrochemical assay for protein kinase activity.

Herein, we report a homogeneous assay for protein kinase activity using an electrochemistry-based probe. The approach involves a peptide substrate conjugated with a redox tag and the phosphate-specific receptor immobilized on an electrode surface. The peptide substrate phosphorylated by a protein kinase binds to the receptor site of the probe, which results in a redox current under voltammetric measurement. Our method was successfully applied even in the presence of citrated human blood and modified to enable a single-use, chip-based electrochemical assay for kinase activity.

Self-assembly behavior of alkylated isophthalic acids revisited: concentration in control and guest-induced phase transformation.

The engineering of two-dimensional crystals by physisorption-based molecular self-assembly at the liquid-solid interface is a powerful method to functionalize and nanostructure surfaces. The formation of high-symmetry networks from low-symmetry building blocks is a particularly important target. Alkylated isophthalic acid (ISA) derivatives are early test systems, and it was demonstrated that to produce a so-called porous hexagonal packing of plane group p6, i.e., a regular array of nanowells, either short alkyl chains or the introduction of bulky groups within the chains were mandatory. After all, the van der Waals interactions between adjacent alkyl chains or alkyl chains and the surface would dominate the ideal hydrogen bonding between the carboxyl groups, and therefore, a close-packed lamella structure (plane group p2) was uniquely observed. In this contribution, we show two versatile approaches to circumvent this problem, which are based on well-known principles: the "concentration in control" and the "guest-induced transformation" methods. The successful application of these methods makes ISA suitable building blocks to engineer a porous pattern, in which the distance between the pores can be tuned with nanometer precision.

Phosphorescent sensor for phosphorylated peptides based on an iridium complex.

A bis[(4,6-difluorophenyl)pyridinato-N,C(2')]iridium(III) picolinate (FIrpic) derivative coupled with bis(Zn(2+)-dipicolylamine) (ZnDPA) was developed as a sensor (1) for phosphorylated peptides, which are related to many cellular mechanisms. As a control, a fluorescent sensor (2) based on anthracene coupled to ZnDPA was also prepared. When the total negative charge on the phosphorylated peptides was changed to -2, -4, and -6, the emission intensity of sensor 1 gradually increased by factors of up to 7, 11, and 16, respectively. In contrast, there was little change in the emission intensity of sensor 1 upon the addition of a neutral phosphorylated peptide, non-phosphorylated peptides, or various anions such as CO3(2-), NO3(-), SO4(2-), phosphate, azide, and pyrophosphate. Furthermore, sensor 1 could be used to visually discriminate between phosphorylated peptides and adenosine triphosphate in aqueous solution under a UV-vis lamp, unlike fluorescent sensor 2. This enhanced luminance of phosphorescent sensor 1 upon binding to a phosphorylated peptide is attributed to a reduction in the repulsion between the Zn(2+) ions due to the phenoxy anion, its strong metal-to-ligand charge transfer character, and a reduction in self-quenching.

Humidity effect of domain wall roughening behavior in ferroelectric copolymer thin films.

We have demonstrated that domain switching in ferroelectric copolymer films can be significantly affected by humidity. With increasing relative humidity (RH), we observed larger domains with highly irregular boundaries as a result of lateral spreading of the tip-induced electric field that originates from water adsorption. Fractal dimension study of irregular domains reveals that the fractal dimension is higher in cases where the RH is higher. The results show that the RH is one of the major switching parameters in ferroelectric copolymers, and therefore could allow clear understanding with regard to domain switching behavior in the ferroelectric copolymer films under ambient conditions.

Photoluminescent and Electrochemiluminescent Detection of Fe3+ Using Cyclometalated Iridium Complexes via Fe3+-Catalyzed Hydrolysis.

Ferric ion (Fe3+) is a biologically abundant and important metal ion. We developed several cyclometalated iridium complex-based molecular sensors (1, ppy-1, 1-phen, 1a, and 1-OMe) for the detection of Fe3+ using an acetal moiety as the reaction site. The acetal moiety in iridium complexes undergoes Fe3+-catalyzed hydrolysis and subsequent formation of a formyl group, resulting in turn-off photoluminescent and electrochemiluminescent responses. Sensor 1 showed excellent selectivity toward Fe3+ over other biologically important metal ions. Furthermore, we compared the performance of the sensors based on the structural differences of the iridium complexes, and revealed a relationship between the structure and chemical properties through electrochemical experiments and computational calculations.

Fluorescent probes designed for detecting human serum albumin on the basis of its pseudo-esterase activity.

We developed activity-based fluorescent probes for detecting human serum albumin (HSA) on the basis of its pseudo-esterase activity. These probes could also detect HSA in blood-contaminated tissue samples.

Reaction-based energy level modulation of a cyclometalated iridium complex for electrochemiluminescent detection of formaldehyde.

Formaldehyde is a toxic compound present in both the environment and living systems, and its detection is important due to its association with various pathological process. In this study, we report a new electrochemiluminescence (ECL) probe based on a cyclometalated iridium complex (IrHAA) for the selective detection of formaldehyde. The homoallylamine moiety in IrHAA reacts with formaldehyde, undergoing a 2-aza-Cope-rearrangement reaction to form a formyl group. Significant changes in the electronic properties and molecular orbital energies of the iridium complex through the functional group transformation result in enhanced ECL and radiometric phosphorescence changes, enabling the quantitative and selective detection of formaldehyde. The energetic requirements for ECL sensing were investigated, highlighting the importance of the excited state energy for achieving efficient ECL. The sensing mechanism was elucidated using NMR spectroscopy and MALDI-TOF analysis.