Probe Compounds Research Articles

ADP-Hep-Induced Liquid Phase Condensation of TIFA-TRAF6 Activates ALPK1/TIFA-Dependent Innate Immune Responses.

The ALPK1 (alpha-kinase 1)-TIFA (TRAF-interacting protein with fork head-associated domain)-TRAF6 signaling pathway plays a pivotal role in regulating inflammatory processes, with TIFA and TRAF6 serving as key molecules in this cascade. Despite its significance, the functional mechanism of TIFA-TRAF6 remains incompletely understood. In this study, we unveil that TIFA undergoes liquid-liquid phase separation (LLPS) induced by ALPK1 in response to adenosine diphosphate (ADP)-β-D-manno-heptose (ADP-Hep) recognition. The phase separation of TIFA is primarily driven by ALPK1, the pT9-FHA domain, and the intrinsically disordered region segment. Simultaneously, TRAF6 exhibits phase separation during ADP-Hep-induced inflammation, a phenomenon observed consistently across various inflammatory signal pathways. Moreover, TRAF6 is recruited within the TIFA condensates, facilitating lysine (K) 63-linked polyubiquitin chain synthesis. The subsequent recruitment, enrichment, and activation of downstream effectors within these condensates contribute to robust inflammatory signal transduction. Utilizing a novel chemical probe (compound 22), our analysis demonstrates that the activation of the ALPK1-TIFA-TRAF6 signaling pathway in response to small molecules necessitates the phase separation of TIFA. In summary, our findings reveal TIFA as a sensor for upstream signals, initiating the LLPS of itself and downstream proteins. This process results in the formation of membraneless condensates within the ALPK1-TIFA-TRAF6 pathway, suggesting potential applications in therapeutic biotechnology development.

Practical examination of flow rate effects and influence of the stationary phase water layer on peak shape and retention in Hydrophilic Interaction Liquid Chromatography.

The effect of flow velocity on retention and peak shape of neutral, acidic and basic probe compounds was studied using seven different UHPLC hydrophilic interaction chromatography (HILIC) columns. Surprisingly on some columns, the retention factor k was found to vary somewhat with flow velocity, due to the combined effects of pressure and of frictional heating on retention. The selectivity of different HILIC columns was much greater than typically found with RP columns. The volume of the water layer on the HILIC columns was measured using the toluene exclusion procedure. For the neutral solute uridine, a good correlation was found between the volume of the water layer and retention, indicating the likely domination of a partition mechanism. For the ionogenic solutes, the correlation was generally poor, due to the presence of strong additional mechanisms such as ionic retention and repulsion. Reduced van Deemter plots for uridine showed a negative correlation between the reduced b coefficient and the volume of the water layer, which can be attributed to reduced surface diffusion in this viscous layer. Once again, the behaviour of ionic solutes was complex on some columns making detailed interpretation difficult.

Enhanced removal of perfluoroalkyl substances using MMO-TiO2 visible light photocatalyst

Exploration of easily produced visible light photocatalysts is still necessary. Thus, this study investigated the catalytic performance of a mixed metal oxide-titanium dioxide (MMO-TiO2) to remove perfluorooctanoic acid (PFOA), a typical and harmful perfluoroalkyl substance, under visible light conditions. Analysis of degradation efficiencies revealed that MMO-TiO2 exhibited a remarkable removal efficiency of 95.99 ± 1.49 %, surpassing other catalysts, including TiO2, ZnAl, MMO, and ZnAl-TiO2. MMO-TiO2, derived from ZnAl-TiO2 calcination, displayed a 5.66-fold higher apparent degradation rate constant (1.07 × 10−2 min−1) compared to ZnAl-TiO2 (1.89 × 10−3 min−1). These materials were characterized via X-ray powder diffractometry, Fourier transform infrared-attenuated total reflectance, N2 adsorption-desorption analysis, and field emission-scanning electron microscopy. Results revealed that O2•- radicals play a pivotal role as the predominant species that mediate PFOA removal. Comparative experiments using probe compounds confirmed increased O2•- production during MMO-TiO2 photoreaction under visible light. In addition, this study explored the effect of operational parameters, such as catalyst dose, initial PFOA concentration, solution pH, and water matrices on the photocatalytic removal process. These findings provide insight into the removal of PFOA using MMO-TiO2, highlighting the potential of this visible light catalyst for the eco-friendly removal of persistent organic pollutants in aquatic systems.

Optimizing radical yield from free chlorine with tailored UV light emitting diode emission spectra

Novel UV sources, which do not contain mercury, provide the opportunity for enhancement of current oxidation technologies through spectral optimization, minimizing inefficiencies that currently limit conventional technology. Wastewater reuse is the primary full-scale application of UV advanced oxidation processes (AOPs) in practice but any background absorbance and the low molar absorption by conventional radical promoters (hydrogen peroxide) have historically limited their system efficiency, resulting in the underutilization of photons in a reactor. This bench-scale research evaluated use of longer wavelength UV light emitting diodes (265, 280, and 300 nm) matched with free chlorine to optimize the utilization of photons for advanced oxidation. Free chlorine possesses large absorption bands in the 280 to 300 nm range in basic pH waters which are common in carbon-based reuse and was used to experimentally verify quantum yields of hydroxyl radical generation across the UV LED peak emission wavelengths. pH- and wavelength-dependent fluence-based rate constants were experimentally derived using Nitrobenzene and Benzoic acid as probe compounds and evaluated to determine the contribution of the hydroxyl and chlorine radical. Reclaimed water taken from various advanced treatment steps was treated with this UV LED AOP to investigate how background absorbance affects radical generation and contaminant transformation kinetics. In addition, alternative performance metrics to evaluate hydroxyl radical production at different incident fluence rates and different rates of photon absorption at unique wavelengths across varying background UV absorbance levels were assessed.

Kinetics and mechanism of dissolved organic phosphorus (DOP) digestion by the UV/O3 process

AbstractIn this study, the kinetics and mechanism of UV/O3 synergistic oxidative digestion of dissolved organic phosphorus (DOP) were investigated, focusing on the ozone direct oxidation and hydroxyl radical oxidation parts of glufosinate and triphenyl phosphate (TPhP). The p‐chlorobenzoic acid (p‐CBA) was selected as the probe compound, and two kinds of reaction kinetic models were proposed by competitive kinetic method with Rct according to the different scale of rate constants of hydroxyl radical oxidation. Under the condition of weakly alkaline (pH = 9.0) and weakly acidic environment (pH = 5.0), the second‐order rate constants of glufosinate and TPhP was determined indirectly to be ko3/glufosinate = (2.903 ± 0.247)M−1s−1 and ko3/TPhP = (3.307 ± 0.204) M−1s−1 by ozone direct oxidation, and k·OH/glufosinate = (1.257 ± 1.031) × 109 M−1s−1 and k·OH/TPhP = (7.120 × 108 ± 0.963) M−1s−1 by hydroxyl radical oxidation, respectively. The comparison of the contribution levels of the two parts to the digestion process showed that the contribution levels in the digestion of glufosinate and TPhP processes both the contribution of ·OH were higher than those of ozone, 86.3% and 72.6%, respectively.

Unraveling the contribution of Co(IV) and free radicals in Co2+-activated Peroxymonosulfate process: Reaction mechanism, degradation pathway, and DFT calculation

The Co2+ and Peroxymonosulfate (PMS) reaction is a highly regarded advanced oxidation process (AOP) that has been extensively researched. The consensus regarding the relative contribution of reactive species in the Co2+/PMS system under different pH conditions has not been established. In this study, quenching experiments and electron spin resonance (ESR) tests demonstrated the coexistence of Co(IV), sulfate radical (SO4•-), hydroxyl radical (•OH), and singlet oxygen (1O2), and there fell to CBZ degradation were Co(IV) > SO4•->•OH ≈ 1O2 in the optimal conditions, where almost 100% of CBZ was degraded in the Co2+/PMS system within 30 min. Furthermore, Cl− would suppress the formation of Co(IV), SO4•-, and •OH but enhance the yield of 1O2, thereby showing a trend of inhibiting CBZ degradation. The probe compound used in this study was Methyl Phenyl Sulfoxide (PMSO), while methyl phenyl sulfone (PMSO2) as the indicator of Co(IV), the generation efficiency of PMSO2 (η'[PMSO2]) increased with reaction time and reached 70%–98% at 40 min at all set pH values (3, 4, 6, and 8). These results suggest that Co(IV) played a significant role in the Co2+/PMS system and its importance became more pronounced as the reaction progressed. PMSO probe and SO4•- quantification experiments jointly revealed that Co(IV) had the greatest performance at pH 4, and compared to other pH values, SO4•- had the largest contribution in the initial stage of the reaction at pH 6. The Ecological Structure Activity Relationships (ECOSAR) software was utilized to predict the toxicity of the degradation by-products. Additionally, luminescent bacteria experiments were conducted to assess the acute toxicity of the reaction solution. The results of these experiments demonstrated that the Co2+/PMS process exhibited a remarkable ability to detoxify the solution.

Changes in the cis-trans isomer selectivity of a reversed-phase liquid chromatography column during use with acidic mobile phase conditions

Reversed-phase liquid chromatography (RPLC) is the analytical tool of choice for monitoring process-related organic impurities and degradants in pharmaceutical materials. Its popularity is due to its general ease-of-use, high performance, and reproducibility in most cases, all of which have improved as the technique has matured over the past few decades. Nevertheless, in our work we still occasionally observe situations where RPLC methods are not as robust as we would like them to be in practice due to variations in stationary phase chemistry between manufactured batches (i.e., lot-to-lot variability), and changes in stationary phase chemistry over time. Over the last three decades several models of RPLC selectivity have been developed and used to quantify and rationalize the effects of numerous parameters (e.g., effect of bonded phase density) on separation selectivity. The Hydrophobic Subtraction Model (HSM) of RPLC selectivity has been used extensively for these purposes; currently the publicly available database of column parameters contains data for 750 columns. In this work we explored the possibility that the HSM could be used to better understand the chemical basis of observed differences in stationary phase selectivity when they occur – for example, lot-to-lot variations or changes in selectivity during column use. We focused our attention on differences and changes in the observed selectivity for a pair of cis-trans isomers of a pharmaceutical intermediate. Although this is admittedly a challenging case, we find that the observed changes in selectivity are not strongly correlated with HSM column parameters, suggesting that there is a gap in the information provided by the HSM with respect to cis-trans isomer selectivity specifically. Further work with additional probe molecules showed that larger changes in cis-trans isomer selectivity were observed for pairs of molecules with greater molecular complexity, compared to the selectivity changes observed for simpler molecules. These results do not provide definitive answers to questions about the chemical basis of changes in stationary phase chemistry that lead to observed differences in cis-trans isomer selectivity. However, the results do provide important insights about the critical importance of molecular complexity when choosing probe compounds and indicate opportunities to develop improved selectivity models with increased sensitivity for cis-trans isomer selectivity.

Quantitative probing of reactive oxygen species and their selective degradation on contaminants in peroxymonosulfate-based process enhanced by picolinic acid

The processes of Fe(III) activated peroxymonosulfate (PMS) in degrading contaminants have been extensively studied. Herein, a biodegradable chelating agent, picolinic acid (PICA), was introduced to the PMS/Fe(III) process to improve the reaction efficiency. The emphases of this study were placed on the quantification of steady-state concentrations of reactive oxygen species (ROS). Experiments presented that five types of ROS, including Fe(IV), SO4•−, HO•, 1O2 and O2•− coexisted in this system. Four typical probe compounds were used to quantify the steady-state concentration of ROS under different variables. The steady-state concentration of Fe(IV) ([Fe(IV)]ss) was 3–5 orders of magnitude higher than that of other ROS, followed by 1O2 and SO4•−, whereas HO• had the lowest concentration. The reaction between PMS and PICA was first explored in our study and results showed that 1O2 and O2•− would form in this reaction. Owing to the hybrid oxidation by multiple ROS, this system showed high oxidation capacity, and could effectively degrade a variety of pollutants. The contributions of ROS to the alleviation of pollutants varied depending on their concentrations and specific reactivity of substrates. Generally, organic contaminants with phenol structures were prone to react with Fe(IV). Overall, this study compared the steady-state concentrations of different ROS and revealed the intrinsic ROS formation mechanisms.

Removing antibiotic resistance genes through the synergistic effect of hydroxyl radical and hypochlorite radical generated in a CeO2@CNT electrified membrane – NaClO system

The removal of antibiotic resistance genes (ARGs) is important for inhibiting the spread of antibiotic resistance in the aquatic environment. In this study, a series of coupled advanced oxidation-disinfection systems were developed based on a carbon nanotube membrane modified with different CeO2 nanocrystals and NaClO (denoted as CeO2@CNT-NaClO system). The removal performance of ARGs by the CeO2@CNT-NaClO systems was tested by treating the simulated water samples containing Pseudomonas sp. HLS-6 with sul1 genes and Escherichia coli TOP10F with tetA genes. The results showed that the CeO2@CNT-NaClO system showed the superior disinfection efficiencies compared to using sole CeO2@CNT electrified membrane and sole NaClO. After treatment by the CeO2@CNT-NaClO system, Pseudomonas sp. HLS-6 and Escherichia coli TOP10F were completely inactivated, and the absolute abundances of sul1 and tetA genes decreased by 4.8 log and 1.8 log, respectively. The probe compounds and quenching experiments showed the synergistic effect of •OH and •ClO is the main contributor to the high removal efficiencies of ARGs by the CeO2@CNT-NaClO system. The rectangular CeO2 particles with (200) and (111) crystal facets exhibited the largest amounts of oxygen vacancy and Lewis acid sites, which favored the coupled system to generate the highest cumulative concentration of •OH and •ClO. This led to a synchronous high-efficiency removal of ARGs and antibiotics in the CeO2@CNT-NaClO system.

Importance of Chain Length in Propagation Reaction on •OH Formation during Ozonation of Wastewater Effluent.

During the ozonation of wastewater, hydroxyl radicals (•OH) induced by the reactions of ozone (O3) with effluent organic matters (EfOMs) play an essential role in degrading ozone-refractory micropollutants. The •OH yield provides the absolute •OH formation during ozonation. However, the conventional "tert-Butanol (t-BuOH) assay" cannot accurately determine the •OH yield since the propagation reactions are inhibited, and there have been few studies on •OH production induced by EfOM fractions during ozonation. Alternatively, a "competitive method", which added trace amounts of the •OH probe compound to compete with the water matrix and took initiation reactions and propagation reactions into account, was used to determine the actual •OH yields (Φ) compared with that obtained by the "t-BuOH assay" (φ). The Φ were significantly higher than φ, indicating that the propagation reactions played important roles in •OH formation. The chain propagation reactions facilitation of EfOMs and fractions can be expressed by the chain length (n). The study found significant differences in Φ for EfOMs and fractions, precisely because they have different n. The actual •OH yield can be calculated by n and φ as Φ = φ (1 + n)/(nφ + 1), which can be used to accurately predict the removal of micropollutants during ozonation of wastewater.

Synthesis and evaluation of curcumin-based near-infrared fluorescent probes for detection of amyloid β peptide in Alzheimer mouse models

The abnormal accumulation of amyloid β protein (Aβ) is one of the most important causes of Alzheimer's disease (AD) and is usually a detecting biomarker. Curcumin and its derivatives have potential Aβ aggregate targeting ability; we synthesized a series of curcumin-based near-infrared fluorescence probes in this study. By characterizing the excitation wavelength and emission wavelength, the imaging characteristics of the investigation in the near-infrared light region were determined; with an increase in the concentration of the probe compounds, the fluorescence intensity showed an upward trend, demonstrating ideal optical characteristics. In vivo, imaging results showed that the synthesized probe compounds could penetrate the blood–brain barrier (BBB) and specifically bind to Aβ in the brain of APP/PS1 mice. Especially for compound 3b, the maximum emission wavelength was around 667 nm, and the fluorescence signal intensity in the brain of the APP/PS1 mice model was more than twice that of the wild control group at 120 min after administration, which could display Aβ pathological changes. The fluorescent probes designed in this study can become an effective tool for early AD diagnosis and visual detection.

The overlooked role of Cr(VI) in micropollutant degradation under solar light irradiation

Hexavalent chromium (Cr(VI)) is ubiquitous in natural environments, whereas its role in the transformation of coexisting contaminants may have been overlooked. In this work, it was reported for the first time that the irradiation of Cr(VI) by solar light (solar light/Cr(VI) system) could effectively degrade various micropollutants with different structures. The removal efficiency of selected micropollutants was increased by 13.3–64.8% by the solar light/Cr(VI) system compared to that by direct solar photolysis. Meanwhile, the oxidation rates were enhanced by 2.2–21.5 folds, while they were negligible by Cr(VI) oxidation alone. Experiments by specific scavengers, probe compounds, fluorescence absorbance, and electron spin resonance analysis demonstrated that hydroxyl radical (•OH) was the major reactive species in the solar light/Cr(VI) system. Further experiments showed that the generation of •OH was closely related to the intermediate Cr(V) generated from Cr(VI) reduction, and Cr(V) could be re-oxidized back to Cr(VI). Increasing solution pH negatively affected model micropollutant (carbamazepine (CBZ)) degradation by the solar light/Cr(VI) system, mainly due to the decreased quantum yield of •OH at higher pH. Coexisting sulfate ions showed negligible effect on CBZ degradation in the solar light/Cr(VI) system, while the presence of bicarbonate, chloride, and humic acid inhibited CBZ degradation to varying degrees, owing to their diverse scavenging effects on •OH. Furthermore, moderate CBZ degradation was also achieved by natural solar light photolysis of Cr(VI). This study demonstrated the pivotal role of Cr(VI) in the transformation of micropollutants under solar irradiation, which advances the understanding of the fate of micropollutants in natural environments.

Expanding Chemical Probe Space: Quality Criteria for Covalent and Degrader Probes.

Within druggable target space, new small-molecule modalities, particularly covalent inhibitors and targeted degraders, have expanded the repertoire of medicinal chemists. Molecules with such modes of action have a large potential not only as drugs but also as chemical probes. Criteria have previously been established to describe the potency, selectivity, and properties of small-molecule probes that are qualified to enable the interrogation and validation of drug targets. These definitions have been tailored to reversibly acting modulators but fall short in their applicability to other modalities. While initial guidelines have been proposed, we delineate here a full set of criteria for the characterization of covalent, irreversible inhibitors as well as heterobifunctional degraders ("proteolysis-targeting chimeras", or PROTACs) and molecular glue degraders. We propose modified potency and selectivity criteria compared to those for reversible inhibitors. We discuss their relevance and highlight examples of suitable probe and pathfinder compounds.

Chromatographic behavior of polar compounds in the transition between reversed-phase and submicellar/micellar liquid chromatography

Micellar liquid chromatography (MLC) is a reversed-phase liquid chromatography (RPLC) mode employing a surfactant, typically sodium dodecyl sulfate (SDS), which is biodegradable, in an aqueous or aqueous-organic mobile phase above its critical micellar concentration (CMC). Under these conditions, substantial alterations in retention, selectivity and peak profile occur in comparison to RPLC using conventional hydro-organic mobile phases. These changes arise from the presence of micelles in the mobile phase and the adsorption of surfactant on the stationary phase. In this work, the chromatographic performance of polar compounds of diverse nature is investigated using mobile phases containing SDS below and above its CMC in the presence of acetonitrile. Three nucleosides, four diuretics, and four sulfonamides, all of biomedical and pharmaceutical significance and possessing distinct acid-base characteristics, were chosen as probe compounds. Modifications in retention and peak shape were investigated using experimental conditions that enable the transition from submicellar to micellar conditions. A comparison was also made with conventional RPLC. Analysing the polar compounds with RPLC under low submicellar or micellar conditions significantly decreases the consumption of organic solvent in comparison to hydrophilic liquid chromatography (HILIC).

Kinetics study of chloride-activated peracetic acid for purifying bisphenol A: Role of Cl2/HClO and carbon-centered radicals

Peracetic acid is an emerging oxidant and disinfectant for wastewater purification. In this study, we first developed a comprehensive and accurate model to elucidate the reaction mechanisms and simulate reaction kinetics of peracetic acid (PAA, CH3C(=O)OOH) activated by chloride (Cl–) based on experimental results and literature. A diversity of experiments methods (e.g., quenching experiments, probe compounds degradation, electron paramagnetic resonance (EPR) measurements) and kinetic modeling were used to determine the reactive species. As a result, carbon-centered radicals and free chlorine reactive species (Cl2 and HClO) were devoted to BPA degradation in the PAA/Cl− system. The carbon-centered radicals CH3C(=O)OO•, CH3C(=O)O•, CH3OO•, and •CH3 greatly accelerated BPA degradation with their corresponding kinetics of kCH3C(=O)OO•, BPA = 2 × 108 M−1 s−1, kCH3C(=O)O•, BPA = 2 × 107 M−1 s−1, k•CH3, BPA = 2 × 106 M−1 s−1 and kCH3OO•, BPA = 2 × 104 M−1 s−1. Dissolved Cl2(l) species was also important for BPA degradation with kCl2, BPA of 2 × 107 M−1 s−1, much higher than HClO/ClO− of kHClO, BPA = 1.2 × 101 M−1 s−1 and kClO−, BPA = 9 × 10−3 M−1 s−1. While free chlorine tends to transform BPA to estrogenic chlorinated organic products, the primary degradation of BPA by carbon-centered radicals results in chlorine-free products, reducing the production of disinfection byproducts during the treatment of saline wastewater. This study improves the knowledge of reaction kinetics and mechanism and reactive species generation in the PAA/Cl– system.

Evaluation of the role of superoxide radical as chain carrier for the formation of hydroxyl radical during ozonation

Superoxide radicals (O2•−) have been suggested as an important chain carrier in the radical chain reaction that promotes ozone (O3) decomposition to hydroxyl radicals (•OH) during ozonation. However, due to the difficulty in measuring transient O2•− concentrations, this hypothesis has not been verified under realistic ozonation conditions during water treatment. In this study, a probe compound was used in combination with kinetic modeling to evaluate the role of O2•− for O3 decomposition during ozonation of synthetic solutions with model promotors and inhibitors (methanol and acetate or tert-butanol) and natural waters (one groundwater and two surface waters). By measurement of the abatement of spiked tetrachloromethane (as a O2•− probe), the O2•− exposure during ozonation was determined. Based on the measured O2•− exposures, the relative contribution of O2•− to O3 decomposition, in comparison to OH−, •OH, and dissolved organic matter (DOM), was quantitatively evaluated using kinetic modeling. The results show that water compositions (e.g., the concentration of promotors and inhibitors, and the O3 reactivity of DOM) have a considerable effect on the extent of the O2•−-promoted radical chain reaction during ozonation. In general, the reaction with O2•− accounted for ∼59‒70% and ∼45‒52% of the overall O3 decomposition during ozonation of the selected synthetic solutions and natural waters, respectively. This confirms that O2•− plays a critical role in promoting O3 decomposition to •OH. Overall, this study provides new insights on the controlling factors for ozone stability during ozonation processes.

Algal organic matter accelerates the photodegradation of tetracycline: Mechanisms, degradation pathways and product toxicity

Algae are ubiquitous in eutrophic surface waters and they play important roles in the sunlight-driven transformation of environmental contaminants. The present study aims to investigate the role of algal organic matter (AOM) separated from Microcystis aeruginosa at different growth stages in tetracycline photodegradation along with the related photosensitive mechanisms. The results showed that AOM, both extracellular organic matter (EOM) and intracellular organic matter (IOM) at different growth phases all promote tetracycline photodegradation, but in different ways and performances. EOM exerts greater photolysis effectiveness than IOM with logarithmic phase (LP) organic matter more effective than that from the algae’s declining growth phase (DP). The promoting efficiencies of tetracycline photodegradation by four AOM types follow the order of LP-EOM > DP-EOM > LP-IOM > DP-IOM, accounting for 1.0420 h−1, 0.8281 h−1, 0.6527 h−1 and 0.4974 h−1, respectively. Such discrepant effectiveness is related to the different chemical constituents and structures of EOM and IOM. Scavenger experiments further demonstrated that triplet-state AOM (3AOM*) contributes the most to tetracycline degradation (30.01%∼48.51%), followed by OH (1.68%∼10.66%) and 1O2 (1.34%∼10.08%). The steady-state concentrations of 3AOM*, OH and 1O2 were measured with probe compounds in the range of 13.950 ∼ 155.234 L/mol, 2.494 × 10−17 ∼ 27.636 × 10−17 M and 1.410 × 10−14 ∼ 26.493 × 10−14 M, respectively. Interestingly, the photolytic intermediates show increasing toxicity to Photobacterium phosphoreum at first which then decreases with the extension of irradiation. These results provide comprehensive insight into the transport and environmental behavior of antibiotics in a eutrophic aqueous environment.

Picolinic Acid-Mediated Catalysis of Mn(II) for Peracetic Acid Oxidation Processes: Formation of High-Valent Mn Species.

Metal-based advanced oxidation processes (AOPs) with peracetic acid (PAA) have been extensively studied to degrade micropollutants (MPs) in wastewater. Mn(II) is a commonly used homogeneous metal catalyst for oxidant activation, but it performs poorly with PAA. This study identifies that the biodegradable chelating ligand picolinic acid (PICA) can significantly mediate Mn(II) activation of PAA for accelerated MP degradation. Results show that, while Mn(II) alone has minimal reactivity toward PAA, the presence of PICA accelerates PAA loss by Mn(II). The PAA-Mn(II)-PICA system removes various MPs (methylene blue, bisphenol A, naproxen, sulfamethoxazole, carbamazepine, and trimethoprim) rapidly at neutral pH, achieving >60% removal within 10 min in clean and wastewater matrices. Coexistent H2O2 and acetic acid in PAA play a negligible role in rapid MP degradation. In-depth evaluation with scavengers and probe compounds (tert-butyl alcohol, methanol, methyl phenyl sulfoxide, and methyl phenyl sulfone) suggested that high-valent Mn species (Mn(V)) is a likely main reactive species leading to rapid MP degradation, whereas soluble Mn(III)-PICA and radicals (CH3C(O)O• and CH3C(O)OO•) are minor reactive species. This study broadens the mechanistic understanding of metal-based AOPs using PAA in combination with chelating agents and indicates the PAA-Mn(II)-PICA system as a novel AOP for wastewater treatment.

Evaluation of Heterogeneous Catalytic Ozonation Process for the Removal of Micropollutants from Water/Wastewater: Application of a Novel Pilot-Scale Continuous Flow System

The present study evaluates the removal of micropollutants from water/wastewater contaminated sources through the application of a heterogeneous catalytic ozonation process, using a pilot-scale continuous operation unit, composed of a membrane module for the diffusion and effective dilution of ozone into the liquid phase to be treated and a plug flow reactor/continuous stirred tank reactor (PFR/CSTR) contact reactor system in series, where the catalyst is recirculated in dispersion mode. The solid materials tested as catalysts are natural and calcined zeolite, Bayoxide and alumina, whereas the examined micropollutants, used in this case as probe compounds, are p-chlorobenzoic acid (p-CBA), atrazine, benzotriazole and carbamazepine. A high-performance liquid chromatography system was used to determine the removal of micropollutants. In the case of p-CBA, an ozone-resistant compound, the addition of catalyst was found to significantly enhance its degradation rate, leading to >99% removal under the optimum defined conditions, i.e., in terms of catalyst concentration, pH, temperature, and process time. On the other hand, in the case of atrazine, a different ozone-resistant compound, the introduction of examined catalysts in the ozonation process was found to reduce the degradation of micropollutant, when compared with the application of single ozonation, indicating the importance of specific affinity between the pollutant and the solid material used as catalyst. Benzotriazole, a moderately ozone-reactive compound was degraded by more than 95% under all experimental conditions and catalysts tested in the pilot unit, while carbamazepine, a highly ozone-reactive compound, was completely removed even during the first stage of treatment process (i.e., at the membrane contactor). When increasing the pH value (in the range 6–8) and the contact time, the performance of catalytic ozonation process also improved.

Characterization of LysoTracker Red uptake by in vitro model cells of the outer blood-retinal barrier: Implication of lysosomal trapping with cytoplasmic vacuolation and cytotoxicity

Lysosomal trapping, a physicochemical process in which lipophilic cationic compounds are sequestered in lysosomes, can affect drug disposition and cytotoxicity. To better understand lysosomal trapping at the outer blood-retinal barrier (BRB), we investigated the distribution of LysoTracker Red (LTR), a probe compound for lysosomal trapping, in conditionally immortalized rat retinal pigment epithelial (RPE-J) cells. LTR uptake by RPE-J cells was dependent on temperature and attenuated by ammonium chloride and protonophore, which decreased the pH gradient between the lysosome and cytoplasm, suggesting lysosomal trapping of LTR in RPE-J cells. The involvement of lysosomal trapping in response to cationic drugs, including neuroprotectants such as desipramine and memantine, was also suggested by an inhibition study of LTR uptake. Chloroquine, which is known to show ocular toxicity, induced cytoplasmic vacuolization in RPE-J cells with a half-maximal effective concentration of 1.35 μM. This value was 59 times lower than the median lethal concentration (= 79.1 μM) of chloroquine, suggesting that vacuolization was not a direct trigger of cell death. These results are helpful for understanding the lysosomal trapping of cationic drugs, which is associated with drug disposition and cytotoxicity in the outer BRB.