Reducing thermal damage to adjacent normal tissue with dual thermo-responsive polymer via thermo-induced phase transition for precise photothermal theranosis.

Photothermal therapy has been extensively studied to improve the light-to-heat efficiency for tumor ablation, but could cause severe damage to adjacent healthy tissue due to the thermal transfer, the random distribution of photothermal agents (PTAs), or combination hereof. Herein, we solve this dilemma with a material design strategy to develop a P(AAm-co-AN)-b-P(NIPAM-co-DMAa)-b-P(AAm-co-AN) ABA triblock copolymer by RAFT polymerization, which exhibits both UCST and LCST dual thermo-responsive behaviors in aqueous solution. The P(AAm-co-AN) block with appropriate AN content allows to finely tune its UCST to ∼ 43°C, which can effectively co-assemble with camptothecin (CPT) and Cy7-TCF, a near-infrared (NIR) PTA, realizing the photo-activated "on-demand" release of CPT and Cy7-TCF. The LCST of P(NIPAM-co-DMAa) segment is adjusted to ∼ 53°C by varying DMAa content, enabling an irreversible sol-to-gel transition. The heat transfer in hydrogel and heat dissipation at the interface of hydrogel-adjacent tissue are limited, resulting in selectively cell killing in tumor, with little hyperthermia in adjacent tissues. Moreover, the hydrogel continues to release CPT to enhance the synergistic efficacy of PTT with chemotherapy. These results suggest that dual thermo-responsive polymer can contribute PTT with high selectivity and negligible side effects for precise medicine. STATEMENT OF SIGNIFICANCE: Photothermal therapy exploits the susceptibility of tumor cells toward external light-induced hyperthermia, but can cause severe damage to adjacent healthy tissue due to thermal transfer, random distribution of photothermal agents (PTAs), or combination hereof. Here, we solve this dilemma by developing a P(AAm-co-AN)-b-P(NIPAM-co-DMAa)-b-P(AAm-co-AN) triblock copolymer with UCST and LCST dual thermo-responsive behaviors, realizing the sequential micelle-unimer-hydrogel phase transitions. The polymer can effectively encapsulate PTA/drug, achieve long systemic circulation, accumulate in tumor through EPR effect, regulate drug release by controlling tumor temperature above UCST via irradiation, and finally exhibit a sol-gel transition, eradicating the heat transfer to adjacent tissue. This represents a practicable strategy to guide the design of next-generation polymeric vector that can contribute PTT with negligible side effects.

Characterization of trichome-specific BAHD acyltransferases involved in acylsugar biosynthesis in Nicotiana tabacum.

Glandular trichomes of tobacco (Nicotiana tabacum) produce blends of acylsucroses that contribute to defence against pathogens and herbivorous insects, but the mechanism of assembly of these acylsugars has not yet been determined. In this study, we isolated and characterized two trichome-specific acylsugar acyltransferases that are localized in the endoplasmic reticulum, NtASAT1 and NtASAT2. They sequentially catalyse two additive steps of acyl donors to sucrose to produce di-acylsucrose. Knocking out of NtASAT1 or NtASAT2 resulted in deficiency of acylsucrose; however, there was no effect on acylsugar accumulation in plants overexpressing NtASAT1 or NtASAT2. Genomic analysis and profiling revealed that NtASATs originated from the T subgenome, which is derived from the acylsugar-producing diploid ancestor N. tomentosiformis. Our identification of NtASAT1 and NtASAT2 as enzymes involved in acylsugar assembly in tobacco potentially provides a new approach and target genes for improving crop resistance against pathogens and insects.

A Novel Chemiluminescent Method for Efficient Evaluation of Heterogeneous Fenton Catalysts Using Cigarette Tar.

The evaluation of the catalytic capacity of catalysts is indispensable research, as catalytic capacity is a crucial factor to dictate the efficiency of heterogeneous Fenton catalysis. Herein, we obtained cigarette tar-methanol extracts (CTME) by applying methanol to cigarette tar and found that CTME could cause CL reactions with Fe2+/H2O2 systems in acidic, neutral, and alkaline media. The CL spectrum experiment indicated that the emission wavelengths of the CTME CL reaction with Fe2+/H2O2 systems were about 490 nm, 535 nm, and 590 nm. Quenching experiments confirmed that hydroxyl radicals (•OH) were responsible for the CL reaction for CTME. Then the CL property of CTME was applied in-situ to rapidly determine the amounts of •OH in tetrachloro-1,4-benzoquinone (TCBQ)/H2O2 system in acidic, neutral and alkaline media, and the CL intensities correlated the best (R2 = 0.99) with TCBQ concentrations. To demonstrate the utility of the CTME CL method, the catalytic capacity of different types and concentrations of catalysts in heterogeneous Fenton catalysis were examined. It was found that the order of CL intensities was consistent with the order of degradation efficiencies of Rhodamine B, indicating that this method could distinguish the catalytic capacity of catalysts. The CTME CL method could provide a convenient tool for the efficient evaluation of the catalytic capacity of catalysts in heterogeneous Fenton catalysis.

Impact of Unadorned Carbon Nitride on Photodegradation and Bioavailability of Multifungicides in the Environment.

Unadorned carbon nitride was synthesized via different nitrogen-rich precursors by thermal polymerization and applied to multifungicides for simultaneous photodegradation in the present study. Urea-derived carbon nitride (UCN) was verified to be most efficient in fungicide removal. The influences of catalyst dosage and pH were studied during the photodegradation process. Hydroxyl radical (•OH) and holes (h+) are the active species during photodegradation of each of the eight fungicides within an aqueous environment. The primary photodegradation products and pathways of all eight fungicides were systematically identified using liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis. In addition, the UCN catalyst was also applied to potted plants. The experimental results revealed that UCN could reduce fungicide residues in plants grown within a contaminated matrix. This study shows promising applications of the UCN catalyst in alleviating the hazards of pesticide residue.

Comparison of Potent Odorants in Raw and Ripened Pu-Erh Tea Infusions Based on Odor Activity Value Calculation and Multivariate Analysis: Understanding the Role of Pile Fermentation.

Infusions prepared from raw pu-erh tea (RAPT) and ripened pu-erh tea (RIPT) showed remarkable aroma differences. Predominant odorants in RAPT and RIPT infusions were identified and compared by the combined use of gas chromatography-olfactometry, aroma extract dilution analysis, odor activity values (OAVs), and multivariate analysis. A total of 35 and 19 odorants (OAV > 1) were detected in RIPT and RAPT, respectively. Odorants in RAPT and RIPT are significantly different in both odor properties and aroma compound intensities. Overall, RAPT contained a complex variety of chemical classes with diverse odors and moderate odor intensities, while RIPT is dominated by structurally and organoleptically similar compounds with high potency. Specifically, stale and musty smelling methoxybenzenes contributed the most to RIPT, while floral-, sweet-, and woody-smelling terpene alcohols, terpene ketones, and phenolic compounds were the predominant odorants in RAPT. Orthogonal partial least squares discriminant analysis revealed that linalool, α-ionone, 1,2,4-trimethoxybenzene, 1,2,3-trimethoxy-5-methylbenzene, 1,2,3,4-tetramethoxybenzene, and 1,2,3-trimethoxybenzene underwent remarkable changes during pile fermentation and could be used as potential odor-active markers for RIPT and RAPT discrimination. The comprehensive aroma characterization of pu-erh tea and determination of the effect of pile fermentation on odorant alteration herein will provide guidance for pu-erh tea flavor quality control and evaluation.

Adsorption behavior of three triazole fungicides on polystyrene microplastics.

Environmental pollution caused by microplastics (MPs) and pesticides has become a global challenge, and increasing evidence shows that MPs can adsorb organic pollutants which may affect their distribution and bioavailability. As widely used pesticides, triazole fungicides with potential environmental and human safety risks often coexist with MPs in the environment. Understanding the adsorption behavior is the basis of risk assessment of co-exposure of MPs and triazole fungicides. In this study, the adsorption behavior of three commonly used triazole fungicides on polystyrene (PS) was studied using adsorption test. The influences of PS particle size and environmental factors on adsorption capacity were evaluated, and the adsorption mechanisms were discussed. Results suggested that the adsorption kinetics and isotherm conformed to the Pseudo-second-order and Freundlich model, respectively. The order of adsorption and desorption capacity was hexaconazole (HEX) > myclobutanil (MYC) > triadimenol (TRI), which was positively correlated with LogKow of pesticides. To a certain extent, the decrease in PS particle size and change in solution pH value and increase in salt ion strength all contribute to increasing adsorption capacity. The main mechanisms of adsorption were hydrophobic and electrostatic interactions. MPs can adsorb and may become the source and sink of triazole fungicides in aqueous environments. Our results demonstrate that more attention should be given to the combined water pollution risk of MPs and triazoles fungicides.

A new method for hydroxyl radical detection by chemiluminescence of flue-cured tobacco extracts.

Chemiluminescence (CL) reactions usually take place in a severely restricted pH regime, thereby confining their application in media at different pH. In this paper, the CL behavior of tobacco-methanol extract (TME) was explored. Surprisingly, TME exhibited CL behavior upon mixing with Fe2+/H2O2, HRP/H2O2 and gold nanoparticles/H2O2 oxidation systems, i.e., in acidic, neutral and alkaline solution respectively. Addition of different reactive oxygen species scavengers verified that the hydroxyl radical (OH) triggers TME CL reactions. Then, the CL behavior of TME was applied to determine OH in tetrachloro-1,4-benzoquinone (TCBQ)/H2O2 system in acidic, neutral and alkaline solutions. CL intensity correlated most strongly (R2 = 0.99) with TCBQ concentration, which was used as a means to indirectly denote OH concentration. This implies that OH could be determined by a TME CL method in a semi-quantitative way regardless of pH value. Therefore, the TME CL method may be a promising method for OH determination in various OH-generating systems.

Enantioselective effects of metalaxyl on soil enzyme activity.

The enantioselective effects of the chiral pesticide metalaxyl on soil enzyme activity were investigated. Incubation experiments were conducted to investigate the effects of metalaxyl enantiomers at different concentrations on the activities of urease, invertase, and catalase as well as the type of activity change (activation vs. inhibition) at different times during incubation. The results indicated that the effects of metalaxyl on the activity of soil enzymes were not only related to the concentration of the enantiomers and soil incubation time, but also to the chiral configuration, suggesting the effects were enantioselective. A pattern of inhibition-recovery-slight stimulation was observed in urease activity of the soil samples treated with metalaxyl enantiomers, but the effects of (-) -R-metalaxyl were stronger than those of (+)-S-metalaxyl at the same concentration. Invertase activity in soil samples treated with metalaxyl enantiomers initially sharply decreased before finally returning to the normal level, and the effects of (+)-S-metalaxyl were stronger than those of (-) -R-metalaxyl at the same concentration. Metalaxyl enantiomers influenced catalase activity in a pattern of slight stimulation-inhibition-recovery, and the effects of (-) -R-metalaxyl were stronger than those of (+)-S-metalaxyl at the same concentration.

Multiresidue determination of 114 multiclass pesticides in flue-cured tobacco by solid-phase extraction coupled with gas chromatography and tandem mass spectrometry.

A sensitive and robust multiresidue method for the simultaneous analysis of 114 pesticides in tobacco was developed based on solid-phase extraction coupled with gas chromatography and tandem mass spectrometry. In this strategy, tobacco samples were extracted with acetonitrile and cleaned up with a multilayer solid-phase extraction cartridge Cleanert TPT using acetonitrile/toluene (3:1) as the elution solvent. Two internal standards of different polarity were used to meet simultaneous pesticides quantification demands in the tobacco matrix. Satisfactory linearity in the range of 10-500 ng/mL was obtained for all 114 pesticides with linear regression coefficients higher than 0.994. The limit of detection and limit of quantification values were 0.02-5.27 and 0.06-17.6 ng/g, respectively. For most of the pesticides, acceptable recoveries in the range of 70-120% and repeatabilities (relative standard deviation) of <11% were achieved at spiking levels of 20, 100, and 400 ng/g. Compared with the reported multiresidue analytical method, the proposed method provided a cleaner test solution with smaller amounts of pigments, fatty acids as well as other undesirable interferences. The development and validation of the high sensitivity, high selectivity, easy automation, and high-throughput analytical method meant that it could be successfully used for the determination of pesticides in tobacco samples.

Rapid and sensitive assay for trantinterol, a novel beta(2)-adrenoceptor agonist, in human plasma using liquid chromatography-tandem mass spectrometry.

A rapid and sensitive assay for trantinterol, a novel beta(2)-adrenoceptor agonist, in human plasma has been developed. Samples containing the analyte and internal standard, clenbuterol, were analyzed by liquid chromatography-tandem mass spectrometry after liquid-liquid extraction with diethyl ether:dichloromethane (60:40, v/v). Separation was performed on a Venusil MP C(18) column (50 mm x 4.6 mm, 5 microm) using methanol:1% formic acid (50:50, v/v) as mobile phase and monitored by multiple reaction monitoring of the precursor-to-product ion transitions of trantinterol at m/z 311.2-->238.1 and clenbuterol at m/z 277.2 --> 203.1. The total run time was only 1.5 min and the method was linear over the concentration range 1-1000 pg/mL with a lower limit of quantitation of 1 pg/mL. Intra- and inter-day precisions (relative standard deviation) were below 7% and 12%, respectively, with accuracy (relative error) below 8%. The method was successfully applied to a pharmacokinetic study involving oral administration of a 50 microg trantinterol tablet to healthy volunteers.

Rapid and sensitive LC-MS/MS assay for the quantitation of 20(S)-protopanaxadiol in human plasma.

This paper describes a rapid and sensitive method for the quantitation of 20(S)-protopanaxadiol (PPD) in human plasma based on high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS). The analyte and internal standard (I.S.), ginsenoside Rh(2), were extracted from plasma by liquid-liquid extraction and separated on a Zorbax extend C(18) analytical column using methanol-acetonitrile-10mM ammonium acetate (47.5:47.5:5, v/v/v) as mobile phase. Detection was by tandem mass spectrometry using electrospray ionization in the positive ion mode and multiple reaction monitoring (MRM). The assay was linear over the concentration range 0.1-100.0ng/ml with a limit of detection of 0.05ng/ml. The method was successfully applied to a clinical pharmacokinetic study in healthy volunteers after a single oral administration of a PPD 25mg capsule.

Quantitation of bergenin in human plasma by liquid chromatography/tandem mass spectrometry.

This paper reports the development and validation of an assay for quantitation of bergenin in human plasma using liquid chromatography/tandem mass spectrometry (LC-MS/MS). Bergenin and the internal standard (I.S.), 5-bromo-2,4(1H,3H)-pyrimidinedione (5-BrU), were separated by reversed phase HPLC and quantitated by MS/MS using electrospray ionization (ESI) and multiple reaction monitoring (MRM) in the negative ion mode. The most intense [M-H](-) MRM transition of bergenin at m/z 326.9-->312.3 was used for quantitation and the transition at m/z 188.9-->42.2 was used to monitor 5-BrU. Stability issues with bergenin required the addition of ascorbic acid to plasma samples prior to storage and analysis within 10 days storage at -80 degrees C. The method was linear in the range 3-1000 ng/mL with intra- and inter-day precision of 3.94-5.96 and 1.62-8.31%, respectively, and accuracy <2.33%. The assay was successfully applied to a pharmacokinetic study in healthy volunteers after administration of a single 250 mg oral dose.

Liquid chromatographic-tandem mass spectrometric method for the simultaneous quantitation of telmisartan and hydrochlorothiazide in human plasma.

A rapid and sensitive method using liquid chromatography-tandem mass spectrometry (LC-MS/MS) has been developed for the simultaneous determination of telmisartan and hydrochlorothiazide in human plasma. Sample preparation involved liquid-liquid extraction with diethyl ether-dichloromethane (60:40, v/v). The analytes and internal standard, probenecid, were separated on a Venusil XBP-C(8) column using gradient elution with acetonitrile-10 mM ammonium acetate-formic acid at a flow rate of 1.2 mL/min. Detection was by electrospray negative ionization mass spectrometry using multiple reaction monitoring of the transitions at m/z 513.0-->469.4 for telmisartan, m/z 295.9-->268.9 for hydrochlorothiazide and m/z 283.9-->239.9 for probenecid. For both analytes, the method was linear in the range 1.00-600 ng/mL with intra- and inter-day precision (as relative standard deviation)