Targeting Hydrogel for Intelligent Recognition and Spatiotemporal Control in Cell-Based Therapeutics.

Smart drug platforms based on spatiotemporally controlled release and integration of tumor imaging are expected to overcome the inefficiency and uncertainty of traditional theranostic modes. In this study, a composite consisting of a thermosensitive hydrogel (polyvinyl alcohol-carboxylic acid hydrogel (PCF)) and a multifunctional nanoparticle (Fe3O4@Au/Mn(Zn)-4-carboxyphenyl porphyrin/polydopamine (FAMxP)) is developed to combine tumor immunogenic cell death (ICD)/immune checkpoint blockade (ICB) therapy under the guidance of magnetic resonance imaging (MRI) and fluorescence imaging (FI). It can not only further recognize the target cells through the folate receptor of tumor cells, but also produce thermal dissolution after exposure to near-infrared light to slowly release FAMxP in situ, thereby prolonging the treatment time and avoiding tumor recurrence. As FAMxP entered the tumor cells, it released FAMx in a pH-dependent manner. Chemodynamic, photothermal and photodynamic therapy can cause significant ICD in cancer cells. ICB can thus be further enhanced by injecting anti-programmed cell death ligand 1, improving the effectiveness of tumor treatment. The developed PCF-FAMxP composite hydrogel may represent an updated drug design approach with simple compositions for cooperative MRI/FI-guided targeted therapeutic pathways for tumors.

Discrimination of polysorbate 20 by high-performance liquid chromatography-charged aerosol detection and characterization for components by expanding compound database and library.

Analyzing polysorbate 20 (PS20) composition and the impact of each component on stability and safety is crucial due to formulation variations and individual tolerance. The similar structures and polarities of PS20 components make accurate separation, identification, and quantification challenging. In this work, a high-resolution quantitative method was developed using single-dimensional high-performance liquid chromatography (HPLC) with charged aerosol detection (CAD) to separate 18 key components with multiple esters. The separated components were characterized by ultra-high-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC-Q-TOF-MS) with an identical gradient as the HPLC-CAD analysis. The polysorbate compound database and library were expanded over 7-time compared to the commercial database. The method investigated differences in PS20 samples from various origins and grades for different dosage forms to evaluate the composition-process relationship. UHPLC-Q-TOF-MS identified 1329 to 1511 compounds in 4 batches of PS20 from different sources. The method observed the impact of 4 degradation conditions on peak components, identifying stable components and their tendencies to change. HPLC-CAD and UHPLC-Q-TOF-MS results provided insights into fingerprint differences, distinguishing quasi products.

An integrated liposome-based microfluidic strategy for rapid colorimetric analysis: A case study of microRNA-21 detection.

In this study, a novel integrated liposome-based microfluidic platform combined with a smartphone was designed for the rapid colorimetric detection of microRNA-21 (miRNA-21) in real samples. The flowing surface-functionalized liposomes were first captured by nucleic acid-functionalized Au nanoparticles in the microfluidic chip. In the presence of miRNA-21, the DNA strand modified on the surface of Au nanoparticles hybridized with the target to form double-stranded products and was cleaved by duplex-specific nuclease (DSN) enzyme, causing the liposomes to be re-released. Then, as the liposomes in the colorimetric module were lysed and the "cellular" contents were released, a step-by-step "glucose-glucose oxidase-3,3',5,5'-tetramethylbenzidine (TMB)" colorimetric reaction process catalyzed by the G-quadruplex/hemin was triggered. The grayscale values were recorded and recognized by the smartphone camera for miRNA-21 analysis. The advantages of the present strategy included the portability of smartphone-based colorimetric assay, the encapsulation and transport of reactants by liposomes and the low solvent usage of microfluidic chip. Under optimal conditions, this assay exhibited a wide linear range from 1 pM to 1 nM (r2 = 0.9981), and the limit of detection of miRNA-21 was as low as 0.27 pM. Moreover, the high specificity of this strategy allowed its successful application to the rapid analysis of miRNA-21 in real blood serum samples of people with type 2 diabetes.

Changes in Concentrations of Polyfluoroalkyl Substances in Human Milk Over Lactation Time and Effects of Maternal Exposure via Analysis of Matched Samples.

Perfluoroalkyl and polyfluoroalkyl substances (PFAS) are potentially related to many adverse health outcomes and could be transferred from maternal blood to human milk, which is an important exposure source for infants during a long-term period. In this study, the maternal blood of 76 women after delivery and their matched human milk samples obtained at 0.5, 1, and 3 months were analyzed by solid-phase extraction method with metal-organic framework/polymer hybrid nanofibers as the sorbents and ultrahigh-performance liquid chromatography-negative electrospray ionization mass spectrometric for quantitative analysis of 31 PFAS. The perfluorooctanoic acid, perfluorooctane sulfonate, and N-methyl perfluorooctane sulfonamido acetic acid (N-MeFOSAA) contributed to more than approximately 50% of the total PFAS concentrations in blood and human milk, while N-MeFOSAA (median: 0.274 ng/mL) was the highest PFAS in human milk at 3 months. The transfer efficiencies for PFAS from maternal blood to human milk at 0.5 months were generally lower, with medians ranging from 0.20% to 16.9%. The number of PFAS species detected in human milk increased as the lactation time went on from 0.5 to 3 months, and the concentrations of 10 PFAS displayed an increasing trend as the prolongation of lactation time (p < 0.05).

Multiplex Detection of Single Nucleotide Polymorphisms by Liquid Chromatography for Nonsmall Cell Lung Cancer Staging.

In this work, an integrated strategy with excellent accuracy and high throughput is proposed for the precise indication of single nucleotide polymorphism (SNP) in nonsmall cell lung cancer diseases. Two types of point mutations (L858R and T790M) and the corresponding wild types could be identified together in a single high-performance liquid chromatographic run. Signal amplification was achieved through a series of enzyme ligation, primer extension, and enzyme cleavage strategies, and a large number of DNA probes with different fluorescence signals were finally generated. The factors affecting the spatiotemporal separation efficiency of four DNA probes were systematically investigated. The limits of detection of wild types (WTs) or mutant types (MTs) abbreviated as L858R-MT, L858R-WT, T790M-MT, and T790M-WT were 26, 24, 19, and 22 aM, respectively. In addition, the levels of mutant types and wild types in the serum of 40 nonsmall cell lung cancer patients at different stages were detected using the method, and the correlation between the mutation ratios and cancer stages was preliminarily verified. The proposed highly selective and sensitive method may serve as an alternative approach for early diagnosis and staging of nonsmall cell lung cancer.

Recent advances in the analysis of electronic cigarette liquids and aerosols: Sample preparation and chromatographic characterization.

Electronic cigarette (e-cigarette) usage has risen dramatically worldwide in recent years. It has been publicized as a safer alternative to the conventional combustible cigarette. This, however, has not yet been supported by robust toxicological research evidence. Analysis of the chemical compositions of e-liquids and generated aerosols is an important step in evaluating the toxicity effects of e-cigarettes. Currently, a broad spectrum of analytical methods have been employed for qualitative and quantitative analysis of chemical compositions of e-cigarette liquids and aerosols. The aim of this article is to review the advances in the chromatographic characterization of chemical composition of the latter in the recent five years. In addition, sample preparation methods for e-liquids and aerosols are surveyed and discussed. A study of the relevant literature indicates that, expectedly, gas chromatography and liquid chromatography with a variety of detection systems, particularly mass spectrometry, have been the main analytical techniques used in this field. Sample preparation procedures primarily include headspace sampling, dilute-and-shoot approach, liquid-liquid extraction and sorbent-based extraction for e-liquids and for aerosols (the latter usually with laboratory-built collection devices). Some challenges of current e-cigarette analytical research, and an overview on prospective work are also presented.

Characteristic and health risk of per- and polyfluoroalkyl substances from cosmetics via dermal exposure.

In this work, 45 cosmetic samples were collected from China, and 27 target per- and polyfluoroalkyl substances (PFAS) were analyzed by ultrahigh-performance liquid chromatography-high resolution mass spectrometry. PFAS were found in all samples, including the products marketed for pregnant women, and the total concentrations of PFAS measured in each sample were in the range of 4.05 - 94.9 ng/g. Short-chain perfluorinated carboxylic acids were the dominant compounds contributing to over 60% of the total content. Perfluorobutanoic acid, with high placental transfer efficiency, was the major PFAS in cosmetics for pregnant women. Three emerging PFAS, 2-perfluorohexyl ethanoic acid, 3-perfluoropentyl propanoic acid (5:3) and perfluoro-2-propoxypropanoic acid, were also identified in the cosmetic samples at quantifiable levels. Significantly, positive correlations between individual PFAS were observed, indicating that there may be a common source for PFAS in these samples. Statistical analyses suggested that using plastic containers and precursor substances may be potential sources of PFAS in terminal products, and product aging may increase PFAS levels. From the PFAS analysis of the cosmetics, the margin of safety (MoS) and hazard quotient (HQ) were calculated to assess human health risks through dermal exposure by using these products. Although the MoS and HQ values obtained were deemed acceptable, the cumulative effect caused by composite and long-term exposure to these contaminants needs to be given greater attention by health authorities.

The sources and bioaccumulation of per- and polyfluoroalkyl substances in animal-derived foods and the potential risk of dietary intake.

Per- and polyfluoroalkyl substances (PFAS) have attracted increasing attention due to their environmental persistence and potential toxicity. Diet is one of the main routes of human exposure to PFAS, particularly through the consumption of animal-derived foods (e.g., aquatic products, livestock and poultry, and products derived from them). This review summarizes the source, bioaccumulation, and distribution of PFAS in animal-derived foods and key influential factors. In most environmental media, perfluorooctanoic acid and perfluorooctane sulfonate are the dominant PFAS, with the levels of short-chain PFAS such as perfluorobutyric acid and perfluorohexane sulfonate surpassing them in some watersheds and coastal areas. The presence of PFAS in environmental media is mainly influenced by suspended particulate matter, microbial communities as well as temporal and spatial factors, such as season and location. Linear PFAS with long carbon chains (C ≥ 7) and sulfonic groups tend to accumulate in organisms and contribute significantly to the contamination of animal-derived foods. Furthermore, PFAS, due to their protein affinity, are prone to accumulate in the blood and protein-rich tissues such as the liver and kidney. Species differences in PFAS bioaccumulation are determined by diet, variances in protein content in the blood and tissues and species-specific activity of transport proteins. Carnivorous fish usually show higher PFAS accumulation than omnivorous fish. Poultry typically metabolize PFAS more rapidly than mammals. PFAS exposures in the processing of animal-derived foods are also attributable to the migration of PFAS from food contact materials, especially those in higher-fat content foods. The human health risk assessment of PFAS exposure from animal-derived foods suggests that frequent consumption of aquatic products potentially engender greater risks to women and minors than to adult males. The information and perspectives from this review would help to further identify the toxicity and migration mechanism of PFAS in animal-derived foods and provide information for food safety management.

Photocatalytic degradation of bisphenol A by temperature-sensitive magnetic hydrogel with enhanced service life.

Easy diffusion and low reusability limit the practicality of photocatalysts. In this study, a hollow sphere (HS) heterojunction was synthesized based on oxygen-doped carbon nitride (OCN) and layered double hydroxides (LDHs). A thermosensitive HS hydrogel (HS Gel) was prepared by mixing HS with N-isopropylacrylamide. Bisphenol A (BPA), being widely manufactured and used in commercial and domestical products and its high toxicity, was chosen as the target pollutant to demonstrate the photocatalytic ability and practicality of the HS Gel. HS Gel presented effective BPA degradation (95% degradation in 70 mins, 4.2 × 10-2 min-1 of kobs) at ambient temperature which is much better than kobs = 1.8 × 10-2 min-1 of OCN and kobs = 0.08 × 10-2 min-1 of LDH), and increased by two-fold the recycling service life (retention of >80% degradation efficiency after 13 usage cycles) compared to other carbon-based photocatalysts (retention of >80% degradation efficiency after 5-6 usage cycles). This is due to its multifunctional characteristics (magnetic property and thermal sensitivity). Under ambient temperature, the hydrophilic HS Gel swelled in the aqueous solution, which promoted the photocatalytic reaction between HS and BPA in the gel state. After the reaction, the HS Gel was subjected to shrinkage by high temperature heating to enhance the mechanical strength for recovery. The magnetic recovery was realized by the paramagnetic properties of layered double oxide to reduce environmental interference. Detailed studies of HS gel related to enhanced service life were conducted including structural changes, catalyst leaking and magnetic changing. A new kind of type Ӏ plus Z-scheme mechanism was also proposed based on the Kubelka-Munk equation, UV diffuse reflectance spectroscopy and Mott-Schotty technique.

Per- and Polyfluoroalkyl Substances in Personal Hygiene Products: The Implications for Human Exposure and Emission to the Environment.

Per- and polyfluoroalkyl substances (PFAS) have been related to reproductive toxicity in humans, but their occurrence in some specific personal hygiene products, i.e., sanitary pads, panty liners, tampons, paper diapers, menstrual cups, and bactericidal liquids, has not been extensively studied. This work investigated 31 representative PFAS in six categories of such personal hygiene products (n = 91). Perfluorinated carboxylic acids were the primary PFAS found in the samples, accounting for over 85% of the total concentrations of PFAS. Paper diapers contained the highest sum of PFAS concentrations (64.6 ng/g) followed by sanitary pads (52.3 ng/g) and menstrual cups (21.1 ng/g). The estimated exposure doses of perfluorooctanoic acid through dermal absorption from the use of menstrual cups and paper diapers for infants (adults) were 0.77 and 2.1 (1.2) ng/kg-bw/day, which contributed more than normal dust ingestion. The estimated emission of paper diapers and sanitary pads into the environment was 2.58 and 322 kg/year with an assumed leaching rate of 100%. The potential exposure of PFAS through the use of personal hygiene products observed in this work suggests a previously unreported exposure pathway of these chemicals to humans.

Application of three dimensional-printed devices in extraction technologies.

Sample pretreatment is an important and necessary process in chemical analysis. Traditional sample preparation methods normally consume moderate to large quantities of solvents and reagents, are time- and labor-intensive and can be prone to error (since they usually involve multiple steps). In the past quarter century or so, modern sample preparation techniques have evolved, from the advent of solid-phase microextraction and liquid-phase microextraction to the present day where they are now widely applied to extract analytes from simple as well as complex matrices leveraging on their extremely low solvent consumption, high extraction efficiency, generally straightforward and simple operation and integration of most, if not all, of the following aspects: Sampling, cleanup, extraction, preconcentration and ready-to-inject status of the final extract. One of the most interesting features of the progress of microextraction techniques over the years lies in the development of devices, apparatus and tools to facilitate and improve their operations. This review explores the application of a recent material fabrication technology that has been receiving a lot of interest, that of three-dimensional (3D) printing, to the manipulation of microextraction. The review highlights the use of 3D-printed devices in the extraction of various analytes and in different methods to address, and improves upon some current extraction (and microextraction) problems, issues and concerns.

Electrospun fluorinated carbon nanotubes/silk fibroin composite nanofibers for the analysis of perfluoroalkyl and polyfluoroalkyl substances.

Assessment of the exposure risk of perfluoroalkyl and polyfluoroalkyl substances (PFASs) has been of public concern for many years. However, it is a challenging undertaking because of the trace levels of these contaminants in the environment and biological systems. In this work, fluorinated carbon nanotubes/silk fibroin (F-CNTs/SF) nanofibers were for the first time synthesized by electrospinning and evaluated as a new adsorbent in pipette tip-solid-phase extraction to enrich PFASs. The addition of F-CNTs increased the mechanical strength and toughness of the SF nanofibers, thus improving the durability of composite nanofibers. The proteophilicity of silk fibroin formed the basis of the good affinity of this material with PFASs. The adsorption behaviors of PFASs on the F-CNTs/SF were investigated by adsorption isotherm experiments to understand the mechanism of extraction. With analysis using ultrahigh performance liquid chromatography-Orbitrap high-resolution mass spectrometric, low limits of detection (0.006-0.090 µg L-1) and enrichment factors of 13-48 were obtained. Meanwhile, the developed method was successfully applied to the detection of wastewater and human placenta samples. This work provides a new idea for the design of novel adsorbents with proteins integrated in polymer nanostructures, a potential routine and practical monitoring technique for PFASs in environmental and biological samples.

Artificial intelligence-assisted smartphone-based sensing for bioanalytical applications: A review.

Artificial intelligence (AI) has received great attention since the concept was proposed, and it has developed rapidly in recent years with applications in many fields. Meanwhile, newer iterations of smartphone hardware technologies which have excellent data processing capabilities have leveraged on AI capabilities. Based on the desirability for portable detection, researchers have been investigating intelligent analysis by combining smartphones with AI algorithms. Various examples of the application of AI algorithm-based smartphone detection and analysis have been developed. In this review, we give an overview of this field, with a particular focus on bioanalytical detection applications. The applications are presented in terms of hardware design, software algorithms, and specific application areas. We also discuss the existing limitations of AI-based smartphone detection and analytical approaches, and their future prospects. The take-home message of our review is that the application of AI in the field of detection analysis is restricted by the limitations of the smartphone's hardware as well as the model building of AI for detection targets with insufficient data. Nevertheless, at this juncture, while bioanalytical diagnostics and health monitoring have set the pace for AI-based smartphone applicability, the future should see the technology making greater inroads into other fields. In relation to the latter, it is likely that the ordinary or average person will play a greater participatory role.

Spatial Confinement of Single-Drop System to Enhance Aggregation-Induced Emission for Detection of MicroRNAs.

Due to high incidence, poor prognosis, and easy transformation into pancreatic cancer (PC) with high mortality, early diagnosis and prevention of acute pancreatitis (AP) have become significant research focuses. In this work, we proposed a magnetic single-drop microextraction (SDME) system with spatial confinement to enhance the aggregation-induced emission (AIE) effect for simultaneous fluorescence detection of miRNA-155 (associated with AP) and miRNA-196a (associated with PC). The target miRNAs were selectively recognized by the hairpin probe and triggered the DNA amplification reaction; then, the DNA strands with two independent probes of G-quadruplex/TAIN and Cy5 were constructed on the surfaces of the magnetic beads. The SDME process, in which a drop containing the fluorescence probes was formed at the tip of the magnetic microextraction rod rapidly within 10 s, was performed by magnetic extraction. In this way, G-quadruplex/TAIN was enriched owing to the spatial confinement of the single-drop system, and the fluorescence signal given off (by G-quadruplex/TAIN) was highly enhanced (AIE effect). This was detected directly by fluorescence spectrophotometry. The approach achieved low limits of detection of 2.1 aM for miRNA-196a and 8.1 aM for miRNA-155 and wide linear ranges from 10 aM to 10 nM for miRNA-196a and from 25 aM to 10 nM for miRNA-155. This novel method was applied to the fluorescence detection of miRNAs in human serum samples. High relative recoveries from 95.6% to 104.8% were obtained.

Self-Generation of Distinguishable Fluorescent Probes via a One-Pot Process for Multiple MicroRNA Detection by Liquid Chromatography.

To address the challenge of signal production and separation for multiple microRNA (miRNA) detection, in this work, a "one-pot" process to self-generate distinguishable fluorescent probes was developed. Based on a long and short probe amplification strategy, the generated G-quadruplex fluorescent dye-free probes can be separated and detected by a high-performance liquid chromatography-fluorescence platform. The free hairpin probes enriched in guanine with different lengths and base sequences were designed and could be opened by the target miRNAs (miRNA-10b, miRNA-21, and miRNA-210). Cleaved G-quadruplex probes with fluorescent signal could be generated in a one-pot process after a duplex-specific nuclease-based cleavage, and the detection of multiple miRNAs could be realized in one run. No solid nanomaterials were applied in the assay, which avoided the blocking of the column. Moreover, without modification of expensive fluorescein, the experimental cost was greatly reduced. The one-pot reaction process also eliminated tedious preparation steps and suggested feasibility of automation. The limits of detection of miRNA-10b, miRNA-21, and miRNA-210 were 2.19, 2.20, and 2.75 fM, respectively. Notably, this method was successfully applied to multiplex detection of miRNAs in serum samples from breast cancer patients within 30 min.

Can Direct-Immersion Aqueous-Aqueous Microextraction Be Achieved When Using a Single-Drop System?

For the analysis of biological analytes in complex matrices, it is difficult to achieve extraction of analytes and enrichment in an aqueous-aqueous single-drop microextraction system. In this study, we proposed a pH-dependent polydopamine (PDA)-coated vesicle/Fe3O4 magnetic aqueous-aqueous in a single-drop microreactor (SDMR) for the direct fluorescence detection of glutathione S-transferase (GST), a metabolic enzyme involved with crucial biological processes, in biological samples. After extracting and enriching the GST target from an aqueous-aqueous single-drop interface, the extraction process was conducted rapidly in 6 s in the SDMR system. The GST was first extracted from the sample solution via the GST-Aptamer on the polydopamine-coated vesicle/Fe3O4 nanospheres (Fe3O4@PDA@GST-Aptamer). Then, as the pH changed from weakly acidic to weakly alkaline in the SDMR system, the GST and GST-Aptamer were released from Fe3O4@PDA@GST-Aptamer nanospheres and captured by polydiacetylene vesicles via the capture probe. These changes altered the effective conjugation length and angle of the vesicle trunk, generating a highly enhanced fluorescence signal. This not only achieved the purpose of target enrichment but also reduced interferences posed by matrix effects. The approach can be used for the direct detection of GST in genuine urine and blood without any sample pretreatment. The linear range was 0.005 to 0.5 µg/mL, and the limit of detection was 0.834 ng/mL. The recoveries of GST in genuine blood samples ranged from 90.8 to 108.0% and in urine from 91.6 to 102.8%. The method has the capability of handling complex samples directly by enabling microextraction in an aqueous-aqueous single-drop system.

Occurrence of Perfluoroalkyl and Polyfluoroalkyl Substances in Ice Cream, Instant Noodles, and Bubble Tea.

Food consumption is a significant exposure route to perfluoroalkyl and polyfluoroalkyl substances (PFAS). The concentrations of 27 PFAS in fast food were determined by ultrahigh-performance liquid chromatography-high resolution mass spectrometry. In ice cream, instant noodles, and bubble tea, some PFAS were detected, among which perfluorooctanoic acid, perfluoro-n-butanoic acid, and 6:2 polyfluoroalkyl phosphate monoester showed relatively high concentrations. PFAS migrating from bubble tea cups to the food simulant of 50% ethanol aqueous solution showed a difference compared with those migrating into bubble tea matrices. The migration of 27 PFAS to bubble tea samples indicated that long storage time increased PFAS levels (up to 4.8 times) and so did high storage temperature (up to 7.3 times). The hazard ratio, defined as the ratio of the estimated daily intake and the reference dose, was calculated, and it suggests that the total PFAS exposure risk due to consumption of bubble tea should be of concern.