Microfluidic paper-based analytical device for measurement of pH using as sensor red cabbage anthocyanins and gum arabic.

The objective of this study was to develop and validate a novel microfluidic paper-based analytical device (µPADpH) for determining the pH levels in foods. Anthocyanins from red cabbage aqueous extract (RCAE) were used as its analytical sensor. Whatman No. 1 filter paper was the most suitable for the device due to its porosity and fiber organization, which allows for maximum color intensity and minimal color heterogeneity of the RCAE in the detection zone of the µPADpH. To ensure the color stability of the RCAE for commercial use of the µPADpH, gum arabic was added. The geometric design of the µPADpH, including the channel length and separation zone diameter, was systematically optimized using colored food. The validation showed that the µPADpH did not differ from the pH meter when analyzing natural foods. However, certain additives in processed foods were found to increase the pH values.

Deep Learning-Based Kinetic Analysis in Paper-Based Analytical Cartridges Integrated with Field-Effect Transistors.

This study explores the fusion of a field-effect transistor (FET), a paper-based analytical cartridge, and the computational power of deep learning (DL) for quantitative biosensing via kinetic analyses. The FET sensors address the low sensitivity challenge observed in paper analytical devices, enabling electrical measurements with kinetic data. The paper-based cartridge eliminates the need for surface chemistry required in FET sensors, ensuring economical operation (cost < $0.15/test). The DL analysis mitigates chronic challenges of FET biosensors such as sample matrix interference, by leveraging kinetic data from target-specific bioreactions. In our proof-of-concept demonstration, our DL-based analyses showcased a coefficient of variation of <6.46% and a decent concentration measurement correlation with an r2 value of >0.976 for cholesterol testing when blindly compared to results obtained from a CLIA-certified clinical laboratory. These integrated technologies have the potential to advance FET-based biosensors, potentially transforming point-of-care diagnostics and at-home testing through enhanced accessibility, ease-of-use, and accuracy.

Effect of incubation conditions of cellulase hydrolysis on mechanical pulp fibre morphology.

The mechanical pulp industry is diversifying through the manufacture of high-value paper products, such as microfibrillated cellulose. However, the development of fibre quality is still energy-intensive. Enzymatic hydrolysis is hypothesized to promote fibre cutting, greater fibrillation, and reduce refining energy costs. Despite potential benefits, there is little understanding of the mechanisms behind fibre development during enzymatic hydrolysis of mechanical pulp. This work investigates how incubation pH and temperature during enzymatic hydrolysis impact the refining of mechanical pulp short fibres. Incubation with endoglucanase at pH 5 and 60 °C increased fibre cutting by approximately 20 %. Fibrillation was negatively affected at this condition, resulting in increased slim fines formation with refining. Incubation at pH 8 and 80 °C promoted >15 % reduction in fibre length, despite such conditions being associated with low enzyme activity. The pH variation modified the sedimentation height of the fibres and the conductivity of suspensions, indicating a change in fibre surface charge. Fibre morphology changes were induced by enzyme hydrolysis conducted at conditions representative of the full range of pH and temperature observed in mechanical pulp mills.

A portable lateral flow distance-based paper sensor for drinking water hardness test.

Hardness is one of the basic parameters of water, and a high-level hardness of drinking water may be harmful to human health. Thus, it is very important to monitor drinking water hardness. In this work, a portable lateral flow distance-based paper sensor for the semi-quantitative detection of drinking water hardness is demonstrated. In the presence of Ca2+/Mg2+, the hydrogel can be formed via the chelation between sodium alginate and Ca2+/Mg2+, inducing a phase separation process. The viscosity change of the sodium alginate solution is directly related to the Ca2+/Mg2+ concentration and can be determined by the water lateral flow distance on test strips. The sensor successfully realizes the quantification of Ca2+ and Mg2+ in the range of 0-10 mmol L-1 and 4-20 mmol L-1, respectively. The recoveries are found varied from 95% to 108.9%. The water hardness is acceptable for drinking if the Cr values lies in the range of 0.259 to 0.419, and it is high with the Cr value above 0.595. Remarkably, the performance of the sensor is comparable with the commercial kit for real water samples, which avoids the subjective judgment. Overall, this method provides a portable approach for semi-quantitative detection of drinking water hardness with the merits of convenience and low cost, which shows great potential for the potential application.

Wearable electrochemical device based on butterfly-like paper-based microfluidics for pH and Na+ monitoring in sweat.

A wearable potentiometric device is reported based on an innovative butterfly-like paper-based microfluidic system, allowing for continuous monitoring of pH and Na+ levels in sweat during physical activity. Specifically, the use of the butterfly-like configuration avoids evaporation phenomena and memory effects, enabling precise and timely biomarker determination in sweat. Two ad hoc modified screen-printed electrodes were embedded in the butterfly-like paper-based microfluidics, and the sensing device was further integrated with a portable and miniaturized potentiostat, leveraging Bluetooth technology for efficient data transmission. First, the paper-based microfluidic configuration was tested for optimal fluidic management to obtain optimized performance of the device. Subsequently, the two electrodes were individually tested to detect the two biomarkers, namely pH and Na+. The results demonstrated highly promising near-Nernstian (0.056 ± 0.002 V/dec) and super-Nernstian (- 0.080 ± 0.003 V/pH) responses, for Na+ and pH detection, respectively. Additionally, several important parameters such as storage stability, interferents, and memory effect by hysteresis study were also investigated. Finally, the butterfly-like paper-based microfluidic wearable device was tested for Na+ and pH monitoring during the physical activity of three volunteers engaged in different exercises, obtaining a good correlation between Na+ increase and dehydration phenomena. Furthermore, one volunteer was tested through a cardiopulmonary test, demonstrating a correlation between sodium Na+ increase and the energetic effort by the volunteer. Our wearable device highlights the high potential to enable early evaluation of dehydration and open up new opportunities in sports activity monitoring.

Fabrication of highly fluorescent graphene quantum dots for quantification of As3+ ion using modified cellulose paper.

Easy, economical, and swift detecting tools are very demanded for assaying various chemical species. The introduction of label-free paper-based read-out devices has significantly reached the demand of analytical science for target analytes assays. Herein, a facile, and disposable inexpensive paper-based sensing tool was fabricated for sensing As3+ ion using graphene quantum dots (GQDs) as a fluorescent reader. The CA-GQDs were synthesized using citric acid (CA) as a precursor via the pyrolysis method, further physisorbed on the cellulose substrate for sensing of As3+ via aggregation-based fluorescence "turn-off" mechanism. The linear range for quantitating As3+ ion is in the range of 0.05-50 µM with a detection limit of 10 nM. The practical application of the CA-GQDs-based analytical platform was verified by assaying As3+ ion in water samples. The CA-GQDs-embedded paper strip can be easily extended for assaying of As3+ ion, which meets the demand for monitoring of As3+ ion in real samples.

Treatment of pulp and paper mill effluent through combined aerobic and anaerobic suspended fixed-bed bioreactor.

This study explored using ultrafiltration (UF) membranes to treat pulp and paper mill wastewater, implementing a novel Taguchi experimental design to optimize operating conditions for pollutant removal and minimal membrane fouling. Researchers examined four factors: pH, temperature, transmembrane pressure, and volume reduction factor (VRF), each at three levels. Optimal conditions (pH 10, 25°C, 6 bar, VRF 3) led to a 35% reduction in flux due to fouling and high pollutant rejections: total hardness (83%), sulfate (97%), spectral absorption coefficient (SAC254) (95%), and chemical oxygen demand (COD) (89%). Conductivity had a lower rejection rate of 50%. Advanced imaging techniques like atomic force microscopy (AFM) and scanning electron microscopy (SEM) revealed reduced membrane fouling under these conditions. The Taguchi method effectively identified optimal conditions, significantly improving wastewater treatment efficiency and promoting environmental sustainability in the pulp and paper industry. PRACTITIONER POINTS: This study optimized UF membrane conditions for pulp and paper mill wastewater, reducing fouling and enhancing pollutant removal, offering practical strategies for industrial treatment. AFM and SEM provided key insights into membrane fouling and mitigation, promoting real-time diagnosis and optimization for enhanced treatment efficiency. Prioritizing anaerobic fixed-bed systems in wastewater treatment is beneficial for achieving high COD removal efficiency. Optimizing hydraulic retention time (HRT) in these systems can further improve their overall effectiveness and sustainability.

Humidity-enhanced microfluidic plasma separation on Chinese Xuan-papers.

The first step in blood testing necessitates blood separation to obtain an adequate volume of plasma. Traditional centrifugation is bulky, expensive and electricity-powered, which is not suitable for micro-scale blood plasma separation in point-of-care testing (POCT) cases. Microfluidic paper-based plasma separation devices present a promising alternative for plasma separation in such occasions. However, they are limited in terms of plasma yield, which hinders analyte detection. Herein, we proposed a humidity-enhanced paper-based microfluidic plasma separation method to address this issue. Specifically, paper was first treated by blood-typing antibodies, then samples of whole blood were introduced into the prepared paper. After waiting for 5 min for RBC agglutination and plasma wicking under high humidity, micro-scale plasma separation from whole blood was achieved. As a result, an extremely high plasma yield of up to 60.1% could be separated from whole blood through using Xuan-paper. Meanwhile, the purity of plasma could reach 99.99%. Finally, this innovative approach was effortlessly integrated into distance-based glucose concentration detection, enabling rapid determination of blood glucose levels through naked-eye observation. Considering the simplicity and inexpensiveness of this method, we believe that this technology could be integrated to more paper-based microfluidic analytical devices for rapid and accurate detection of plasma analytes in POCT.

Pancreatic lipase immobilization on cellulose filter paper for inhibitors screening and network pharmacology study of anti-obesity mechanism.

The discovery of pancreatic lipase (PL) inhibitors is an essential route to develop new anti-obesity drugs. In this experiment, chitosan was used to add amino groups to cellulose filter paper (CFP) and then glutaraldehyde was used to covalently combine PL with amino-modified CFP through the Schiff base reaction. Under optimal immobilization conditions, CFP immobilized PL has a wide range of pH and temperature tolerance, as well as excellent reproducibility, reusability and storage stability. Subsequently, 26 natural products (NPs) were screened by immobilized PL with black tea extract having the highest inhibition rate. Three compounds with binding effects on PL (epigallocatechin gallate, theaflavin-3-gallate and theaflavin-3,3'-digallate) were captured. Molecular docking proved that these three compounds have a strong binding affinity for PL. Fluorescence spectra further revealed that theaflavin-3,3'-digallate could statically quench the intrinsic fluorescence of pancreatic lipase. The molecular docking and thermodynamic parameters indicated that electrostatic interaction was considered as the main interaction force between PL and theaflavin-3,3'-digallate. Finally, the potential anti-obesity targets and pathways of the three compounds were discussed through network pharmacology. This study not only proposes a simple and efficient method for screening PL inhibitors, but also sheds light on the anti-obesity mechanism of active compounds in black tea.

Thermal crosslinking kinetics of shellac and its coating for stiffened and water stable cellulose-based paper straws.

In this work, shellac and its crosslinking were studied to produce paper straws for the application of liquid products. Commercial paper straws are not durable for liquid foods due to their hygroscopic nature, and thus, they find it challenging to replace single-use plastics. Shellac is a naturally occurring resin utilized as an adhesive and water-resistant coating over the paper straw. Shellac was cured at 125 °C, 150 °C, 175 °C, and 200 °C, and it was crosslinked in about 210 min, 150 min, 60 min, and 30 min respectively and studied for kinetics. The crosslinking of shellac produced a thermally stable material. Compared to commercial paper straws, these paper shellac straws exhibited high bending stiffness (1356.11 Nmm), tensile strength (13,74 MPa), flexural strength (21.72 MPa), and compression strength (24.99 MPa). Moreover, the paper shellac straws didn't bend in wet conditions under load for up to one day, while the commercial paper straw bends in 8 min. Therefore, paper straws with shellac can replace plastic-based straws for a sustainable future.

Innovative disposable in-tip cellulose paper (DICP) device for facile determination of pesticides in postmortem blood samples: A proof-of-concept study.

Accurately identifying and quantifying toxicants is crucial for medico-legal investigations in forensic toxicology; however, low analyte concentrations and the complex samples matrix make this work difficult. Therefore, a simplified sample preparation procedure is crucial to streamline the analysis to minimize sample handling errors, reduce cost and improve the overall efficiency of analysis of toxicants. To address these challenges, an innovative disposable in-tip cellulose paper (DICP) device has been developed for the extraction of three pesticides viz. Chlorpyrifos, Quinalphos and Carbofuran from postmortem blood samples. The DICP device leverages cellulose paper strips housed within a pipette tip to streamline the extraction process, significantly reducing solvent usage, time, and labor while maintaining high analytical accuracy. The extraction of pesticides from postmortem blood using the DICP device involves a streamlined process characterized by adsorption and desorption. The diluted blood samples were processed through the DICP device via repeated aspirating and dispensing calyces to adsorb the pesticides onto the cellulose paper. The adsorbed pesticides are then eluted using acetone, which is collected for GC-MS analysis. The method was meticulously optimized, achieving a limit of quantification in the range of 0.009-0.01 µg mL-1. The intra-day and inter-day precisions were consistently less than 5 % and 10 %, respectively, with accuracy ranging from 94-106 %. Relative recoveries for the analytes were observed to be between 60 % and 93.3 %, and matrix effects were determined to be less than 10 %. The method's sustainability was validated with a whiteness score of 98.8, an AGREE score of 0.64, a BAGI score of 70 and ComplexMoGAPI score of 77. Applicability was demonstrated through successful analysis of real postmortem blood samples and proficiency testing samples, highlighting its potential utility in forensic toxicology.

Genetically engineered bacterial biofilm materials enhances portable whole cell sensing.

In recent years, whole-cell biosensors (WCBs) have emerged as a potent approach for environmental monitoring and on-site analyte detection. These biosensors harness the biological apparatus of microorganisms to identify specific analytes, offering advantages in sensitivity, specificity, and real-time monitoring capabilities. A critical hurdle in biosensor development lies in ensuring the robust attachment of cells to surfaces, a crucial step for practical utility. In this study, we present a comprehensive approach to tackle this challenge via engineering Escherichia coli cells for immobilization on paper through the Curli biofilm pathway. Furthermore, incorporating a cellulose-binding peptide domain to the CsgA biofilm protein enhances cell adhesion to paper surfaces, consequently boosting biosensor efficacy. To demonstrate the versatility of this platform, we developed a WCB for copper, optimized to exhibit a discernible response, even with the naked eye. To confirm its suitability for practical field use, we characterized our copper sensor under various environmental conditions-temperature, salinity, and pH-to mimic real-world scenarios. The biosensor-equipped paper discs can be freeze-dried for deployment in on-site applications, providing a practical method for long-term storage without loss of sensitivity paper discs demonstrate sustained functionality and viability even after months of storage with 5 µM limit of detection for copper with visible-to-naked-eye signal levels. Biofilm-mediated surface attachment and analyte sensing can be independently engineered, allowing for flexible utilization of this platform as required. With the implementation of copper sensing as a proof-of-concept study, we underscore the potential of WCBs as a promising avenue for the on-site detection of a multitude of analytes.

Micro-macro SERS strategy for highly sensitive paper cartridge with trace-level molecular detection.

Surface-enhanced Raman Scattering (SERS) has become a powerful spectroscopic technology for highly sensitive detection. However, SERS is still limited in the lab because it either requires complicated preparation or is limited to specific compounds, causing poor applicability for practical applications. Herein, a micro-macro SERS strategy, synergizing polymer-assisted printed process with paper-tip enrichment process, is proposed to fabricate highly sensitive paper cartridges for sensitive practical applications. The polymer-assisted printed process finely aggregates nanoparticles with a discrete degree of 1.77, and SERS results are matched with theoretical enhancement, indicating small cluster-dominated hotspots at the micro-scale and thus 41-fold SERS increase compared to other aggregation methods. The paper-tip enrichment process moves molecules in a fluid into small tips filled with plasmonic clusters, and molecular localization at hotspots is achieved by the simulation and optimization of fluidic velocity at the macro-scale, generating a 39.5-fold SERS sensibility increase in comparison with other flow methods. A highly sensitive paper cartridge contains a paper-tip and a 3D-printed cartridge, which is simple, easy-to-operate, and costs around 2 US dollars. With a detection limit of 10 -12 M for probe molecules, the application of real samples and multiple analytes achieves single-molecule level sensitivity and reliable repeatability with a 30-min standardized procedure. The micro-macro SERS strategy demonstrates its potential in practical applications that require point-of-care detection.

Competitive coordination assisted scalable fabrication of FITC­nickel frameworks anchored nanofiber paper for colorimetric/fluorescent monitoring of shrimp freshness.

A novel type of colorimetric/fluorescent nanopaper indicator has been developed from the melt-extruded poly (vinyl alcohol-co-ethylene) nanofibers with surface anchored metal-organic frameworks (MOFs) by an interfacial coordination strategy. Specifically, the fluorescein isothiocyanate molecules could be anchored to the nanofiber surface by nickel ions and co-assembled into a hydrophilic nanocoating via a dynamic water/alcohol solvent evaporation method. Interestingly, this hydrophilic surface enables fast adsorption of moistures and interaction with biological amine vapors, resulting a saffron cake-layer of MOF nanocrystals with ultra-sensitive colorimetric/fluorescent responses based on an alkaline pH/ammonia induced competitive coordination mechanism. Finally, these porous nanofibrous matrix and active nanocoating make the nano-paper an ultra-sensitive optical platform for in-situ monitoring of the shrimp freshness from mins to weeks. Therefore, this composite film shows great potential into advanced paper-based indicators for food quality control and safety in processing industry.

Multi-Functional Repair and Long-Term Preservation of Paper Relics by Nano-MgO with Aminosilaned Bacterial Cellulose.

Paper relics, as carrieres of historical civilization's records and inheritance, could be severely acidic and brittle over time. In this study, the multi-functional dispersion of nanometer magnesium oxide (MgO) carried by 3-aminopropyl triethoxysilane-modified bacterial cellulose (KH550-BC) was applied in the impregnation process to repair aged paper, aiming at solving the key problems of anti-acid and strength recovery in the protection of ancient books. The KH550-BC/MgO treatment demonstrated enhanced functional efficacy in repairing aged paper, attributed to the homogeneous and stable distribution of MgO within the nanofibers of BC networks, with minimal impact on the paper's wettability and color. Furthermore, the treatment facilitated the formation of adequate alkali reserves and hydrogen bonding, resulting in superior anti-aging properties in the treated paper during prolonged preservation. Even after 30 days of hygrothermal aging tests, the paper repaired by KH550-BC/MgO was still in a gently alkaline environment (pH was about 7.56), alongside a 32.18% elevation compared to the untreated paper regarding the tear index. The results of this work indicate that KH550-BC/MgO is an effective reinforcement material for improving the long-term restoration of ancient books.

Waste newspaper as cellulose resource of activated carbon by sodium salts for methylene blue and congo red removal.

The work was aimed at evaluating the adsorptive properties of waste newspaper (WN) activated carbons chemically produced using sodium salts for methylene blue (MB) and congo red (CR) removal. The activated carbons, designated as AC1, AC2, AC3 and AC4 were prepared through impregnation with NaH2PO4, Na2CO3, NaCl and NaOH, respectively and activation at 500 °C for 1 h. The activated carbons were characterized for surface chemistry, thermal stability, specific area, morphology and composition. The AC1 with a surface area of 917 m2/g exhibits a greater MB capacity of 651 mg/g. Meanwhile, a greater CR capacity was recorded by AC2 at 299 mg/g. The pseudo-second order model fitted well with the kinetic data, while the equilibrium data could be described by Langmuir model. The thermodynamic parameters, i.e.., positive ΔH°, negative ΔG° and positive ΔS° suggest that the adsorption of dyes is endothermic, spontaneous and feasible at high solution temperature. To conclude, WN is a potential cellulose source for producing activated carbon, while NaH2PO4 activation could be employed to convert WN into activated carbon for effective dye wastewater treatment.

Unconventional strategies for liver tissue engineering: plant, paper, silk and nanomaterial-based scaffolds.

The paper highlights how significant characteristics of liver can be modeled in tissue-engineered constructs using unconventional scaffolds. Hepatic lobular organization and metabolic zonation can be mimicked with decellularized plant structures with vasculature resembling a native-hepatic lobule vascular arrangement or silk blend scaffolds meticulously designed for guided cellular arrangement as hepatic patches or metabolic activities. The functionality of hepatocytes can be enhanced and maintained for long periods in naturally fibrous structures paving way for bioartificial liver development. The phase I enzymatic activity in hepatic models can be raised exploiting the microfibrillar structure of paper to allow cellular stacking creating hypoxic conditions to induce in vivo-like xenobiotic metabolism. Lastly, the paper introduces amalgamation of carbon-based nanomaterials into existing scaffolds in liver tissue engineering.

Unconventional scaffolds have the potential to meet the current challenges in liver tissue engineering- loss of hepatic morphology and functions over long-term culture, absence of native-like cell-cell and cell-matrix interactions, organization of hepatocytes into lobular structures exhibiting metabolic variations-which hinder pharmaceutical analysis, regenerative therapies and artificial organ development. Paper with cellulose microfibril network develops cellular aggregates with hypoxic conditions that influence enzymes of xenobiotic metabolism proving to be a better scaffold for hepatotoxicity testing compared with conventional monolayers in tissue culture plates. Decellularized plant stems provide already-built vasculature to be exploited for the development of intricate vessel networks that exist in hepatic lobules aiding in regenerative medicine for hepatic pathologies. Fibrous plant structures are excellent materials for the immobilization of hepatocytes and improve albumin secretion enabling their use in bioartificial liver development. Biomimicry of metabolic zonation in hepatic lobules can be achieved with perfusion culture using silk blend scaffolds with varying proportions of the liver matrix that orchestrate cellular function. The mechanical properties of silk allow the fabrication of structures that resemble liver anatomy to generate native-like hepatic lobules. Nanomaterials have immense potential as a component of composite material development for scaffolds to achieve improved predictive ability in pharmacokinetics. Most of these unconventional scaffolds have the added advantage of being readily available, accessible, affordable and sustainable for liver tissue engineering applications. Conclusively, the shift of attention away from conventional scaffolds poses a promising future in the field of tissue engineering.

Molecularly Imprinted Wearable Sensor with Paper Microfluidics for Real-Time Sweat Biomarker Analysis.

The urgent need for real-time and noninvasive monitoring of health-associated biochemical parameters has motivated the development of wearable sweat sensors. Existing electrochemical sensors show promise in real-time analysis of various chemical biomarkers. These sensors often rely on labels and redox probes to generate and amplify the signals for the detection and quantification of analytes with limited sensitivity. In this study, we introduce a molecularly imprinted polymer (MIP)-based biochemical sensor to quantify a molecular biomarker in sweat using electrochemical impedance spectroscopy, which eliminates the need for labels or redox probes. The molecularly imprinted biosensor can achieve sensitive and specific detection of cortisol at concentrations as low as 1 pM, 1000-fold lower than previously reported MIP cortisol sensors. We integrated multimodal electrochemical sensors with an iontophoresis sweat extraction module and paper microfluidics for real-time sweat analysis. Several parameters can be simultaneously quantified, including sweat volume, secretion rate, sodium ion, and cortisol concentration. Paper microfluidic modules not only quantify sweat volume and secretion rate but also facilitate continuous sweat analysis without user intervention. While we focus on cortisol sensing as a proof-of-concept, the molecularly imprinted wearable sensors can be extended to real-time detection of other biochemicals, such as protein biomarkers and therapeutic drugs.

A dual colorimetric-electrochemical microfluidic paper-based analytical device for point-of-care testing of ischemic strokes.

A novel microfluidic paper-based analytical device with dual colorimetric and electrochemical detection (dual µPAD) was developed for the assessment of transferrin saturation (TSAT) in samples from ischemic stroke patients. TSAT was calculated from the ratio between transferrin-bound iron, which was colorimetrically measured, and the total iron-binding capacity, which was electrochemically measured. To this end, a µPAD was smartly designed, which integrated both colorimetric and electrochemical detection reservoirs, communicating via a microchannel acting as a chemical reactor, and with preloading/storing capabilities (reagent-free device). This approach allowed the dual and simultaneous determination of both parameters, providing an improvement in the reliability of the results due to an independent signal principle and processing. The µPADs were validated by analyzing a certified reference material, showing excellent accuracy (Er ≤ 5%) and precision (RSD ≤ 2%). Then they were applied to the analysis of diagnosed serum samples from ischemic stroke patients. The results were compared to those provided by a free-interference method (urea-PAGE). Impressively, both methods exhibited a good correlation (r = 0.96, p < 0.05) and no significant differences were found between them (slope 1.0 ± 0.1 and the intercept 1 ± 4, p < 0.05), demonstrating the excellent accuracy of our approach during the analysis of complex samples from ischemic stroke patients, using just 90 µL of clinical samples and taking less than 90 min in comparison with the 18 hours required by the urea-PAGE approach. The developed fully integrated colorimetric-electrochemical µPAD is a promising ready to use reagent-free device for the point-of-care testing of TSAT, which can be used to assist physicians in the fast diagnosis and prognosis of ischemic strokes, where the decision-time is crucial for the patient's survival.

Distance-based paper device coupled with uracil-rich DNA hydrogel for visual quantification of Uracil-DNA glycosylase.

Uracil-DNA glycosylase (UDG), an enzyme for repairing uracil-containing DNA damage, is crucial for maintaining genomic stability. Simple and fast quantification of UDG activity is essential for biological assay and clinical diagnosis, since its aberrant level is associated with DNA damage and various diseases. Herein, we developed a fully integrated "sample in-signal out" distance-based paper analytical device (dPAD) for visual quantification of UDG using a flow-controlled uracil-rich DNA hydrogel (URDH). The uracil base sites contained in the DNA hydrogel are mis-incorporated with dUTP by rolling circle amplification (RCA), which simplifies the preparation process of the functionalized hydrogel. In the presence of UDG, the uracil in URDH can be recognized and removed to induce the permeability change of URDH, resulting in the visible distance signal along the paper channel. Using dPAD, as low as 6.4 × 10-4 U/mL of UDG (within 80 min) is visually identified without any instruments and complicated operations. This integrated dPAD is advantageous for its simplicity, cost effectiveness, and ease of use. We envision that it has the great potential for point-of-care testing (POCT) in DNA damage testing, personalized healthcare assessment, and biomedical applications.