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Here, we describe a SERS-based vertical flow assay as a platform technology suitable for point-of-care (POC) diagnostic testing. A capture substrate is constructed from filter paper embedded with spherical gold nanoparticles (AuNPs) and functionalized with an appropriate capture antibody. The capture substrate is loaded into a filtration device and connected to a syringe to rapidly and repeatedly pass the sample through the sensor for efficient antigen binding. The antigen is then labeled with a SERS-active detection probe. We show that only a few Raman reporter molecules, exclusively located adjacent to the plasmonic capture substrate, generate detectible signals. To maximize the signal from underutilized Raman reporter molecules, we employ a secondary signal enhancing probe that undergoes antibody-directed assembly to form plasmonic core-satellites. This facile enhancement step provides a 3.5-fold increase in the signal and a detection limit of 0.23 ng/mL (1.6 pM) for human IgG. This work highlights the potential to rationally design plasmonic architectures using widely available and reproducible spherical AuNPs to achieve large SERS enhancements for highly sensitive POC diagnostics.
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Ouro , Imunoglobulina G , Nanopartículas Metálicas , Análise Espectral Raman , Análise Espectral Raman/métodos , Ouro/química , Nanopartículas Metálicas/química , Imunoensaio/métodos , Humanos , Imunoglobulina G/imunologia , Limite de Detecção , Anticorpos/imunologia , Anticorpos/químicaRESUMO
Various chimeric lysins have been developed as efficacious antibiotics against multidrug-resistant bacteria, but direct comparisons of their antibacterial activities have been difficult due to the preparation of multiple recombinant chimeric lysins. Previously, we reported an Escherichia coli cell-free expression method to better screen chimeric lysins against Staphylococcus aureus, but we still needed to increase the amounts of expressed proteins enough to be able to detect them non-isotopically for quantity comparisons. In this study, we improved the previous cell-free expression system by adding a previously reported artificial T7 terminator and reversing the different nucleotides between the T7 promoter and start codon to those of the T7 phage. The new method increased the expressed amount of chimeric lysins enough for us to detect them using Western blotting. Therefore, the qualitative comparison of activity between different chimeric lysins has become possible via the adjustment of the number of variables between samples without protein purification. We applied this method to select more active chimeric lysins derived from our previously reported chimeric lysin (ALS2). Finally, we compared the antibacterial activities of our selected chimeric lysins with reported chimeric lysins (ClyC and ClyO) and lysostaphin and determined the rank orders of antibacterial activities on different Staphylococcus aureus strains in our experimental conditions.
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Antibacterianos , Bacteriófagos , Antibacterianos/farmacologia , Staphylococcus aureus/metabolismo , Lisostafina , N-Acetil-Muramil-L-Alanina Amidase , Bacteriófagos/metabolismoRESUMO
Validating the direct photocatalytic activity of colloidal plasmonic nanoparticles is challenging due to their limited stability and needed support materials that can often contribute to the chemical reactions. Stable gold nanoparticles (AuNPs) with tunable sizes are prepared across porous polymer particles without any chemical bonds where the resulting composite particles exhibit intense surface plasmon resonances (SPRs) in the visible region. These composite particles are then tested as photocatalysts under a broadband solar-simulated light source to examine the contribution degree of photothermal heating and SPR coming from the incorporated AuNPs in the C-C bond forming homocoupling reaction. Generally, the thermal and photothermal heating are the main driving force to increase the reactivity of relatively smaller AuNPs (~ 44 nm in diameter) with a narrower SPR band. However, the SPR-induced catalytic activity is much greater for the composite particles containing larger AuNPs (~ 87 nm in diameter) with a broader SPR. As the polymer particle matrix does not influence the catalytic activity (e.g., inducing charge delocalization and/or separation), the unique SPR role of the colloidal AuNPs in the catalytic reaction is assessable under light irradiation. This study experimentally demonstrates the possibility of evaluating the direct contribution of SPRs to photocatalytic chemical reactions.
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Rapid, sensitive, and quantitative detection of biomarkers is needed for early diagnosis of disease and surveillance of infectious outbreaks. Here, we exploit a plasmonic syringe filter and surface-enhanced Raman spectroscopy (SERS) in the development of a rapid detection system, using human IgG as a model diagnostic biomarker. The novel assay design facilitates multiple passages of the sample and labeling solution through the detection zone enabling us to investigate and maximize sampling efficiency to the capture substrate. The vertical flow immunoassay process in this study involves the utilization of filter paper embedded with gold nanoparticles (AuNPs) to form a plasmonic substrate. Capture antibody (anti-human IgG) is then immobilized onto the prepared plasmonic paper and inserted into a vertical flow device (syringe filter holder). Sample solution is passed through the filter paper and the target antigen (human IgG) is selectively captured by the immobilized antibody to form an antibody-antigen complex. Next, functionalized AuNPs as extrinsic Raman labels (ERLs) are passed through the filter paper to label the captured biomarker molecules forming a layered structure. This sandwiched geometry enhances plasmonic coupling and SERS signal to provide highly sensitive detection of biomolecules. Systematic studies to investigate the impact of multiple infuse/withdraw cycles of the sample and labeling solutions reveal that antigen and ERL binding are maximized with 10 and 20 cycles, respectively. The optimized assay achieves a detection limit of â¼0.2 ng mL-1 for human IgG with a total assay time of less than 5 minutes, meeting the demands for rapid point of care diagnostics. Additionally, the optimized platform was implemented in the quantitative analysis of the SARS-CoV-2 nucleocapsid protein, the typical target in commercial, FDA-approved rapid antigen tests for COVID-19.
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Ouro , Nanopartículas Metálicas , Humanos , Ouro/química , Nanopartículas Metálicas/química , Análise Espectral Raman/métodos , Biomarcadores , Imunoensaio/métodos , Antígenos , Imunoglobulina GRESUMO
This work demonstrates the incorporation of gold nanoparticles (AuNPs) into crosslinker-free poly(N-isopropylacrylamide), PNIPAM, particles in situ and the examination of their structural and catalytic properties. The formation process of the AuNPs across the crosslinker-free PNIPAM particles are compared to that of crosslinked PNIPAM particles. Given the relatively larger free volume across the crosslinker-free polymer network, the AuNPs formed by the in situ reduction of gold ions are detectably larger and more polydisperse, but their overall integration efficiency is slightly inferior. The structural features and stability of these composite particles are also examined in basic and alcoholic solvent environments, where the crosslinker-free PNIPAM particles still offer comparable physicochemical properties to the crosslinked PNIPAM particles. Interestingly, the crosslinker-free composite particles as a colloidal catalyst display a higher reactivity toward the homocoupling of phenylboronic acid and reveal the importance of the polymer network density. As such, the capability to prepare composite particles in a controlled polymer network and reactive metal nanoparticles, as well as understanding the structure-dependent physicochemical properties, can allow for the development of highly practical catalytic systems.
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Oblique lumbar interbody fusion is a minimally invasive procedure for treating degenerative lumbar disease. Its advantages include correcting coronal and sagittal spinal alignment and indirect neural decompression. However, achieving a successful outcome is limited in some patients who need direct decompression for central canal lesions including hard stenotic lesions (endplate or facet articular osteophytes and ossification of posterior longitudinal ligaments) and sequestration of the disk. Biportal endoscopic spinal surgery is a minimally invasive technique, which directly decompresses the lesion. By taking advantage of two procedures, in a longlevel lumbar lesion, alignment correction and direct decompression can be both achieved. Herein, the authors introduce multilevel lumbar fusion through oblique lumbar interbody fusion and selective direct decompression through biportal endoscopic spinal surgery and discuss the surgical indications, surgical pitfalls, and recommendations for application. Consequently, it is regarded as a minimally invasive interbody fusion method for patients with multilevel lumbar degenerative degeneration.
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This study reports the effects of recovered carbon black (produced in a clean and sustainable way) as a reinforcing agent on the physicochemical properties of a styrene-butadiene rubber (SBR) matrix. SBR-based composite materials are prepared with recovered green carbon black (GCB), and these are thoroughly compared to the composite materials containing conventional virgin carbon black (VCB) (produced by the incomplete combustion of petroleum products). The GCB-SBR composite materials generally show detectably inferior properties compared to the VCB-SBR composite under the same preparation conditions due to the limited functionality of the GCB filler. However, the introduction of a small amount of crosslinker, acrylate-functionalized POSS (polyhedral oligomeric silsesquioxane), into the GCB-SBR composite materials effectively enhances the overall physical properties, including the tensile strength, fracture elongation, and thermal stability. The degree of the crosslinking efficiency, thermal stability, and mechanical properties of the composite materials are optimized and thoroughly examined to demonstrate the possibility of replacing typical VCB with GCB, which can allow for upcycling the inexpensive and ecofriendly carbon black materials as effective reinforcing fillers.
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Objectives: Sarcopenia is a skeletal muscle loss disease with adverse outcomes, including falls, mortality, and cardiovascular disease (CVD) in older patients. Distal radius fractures (DRF), common in older people, are strongly related to falls. We aimed to investigate the correlation between DRF and low skeletal muscle mass, which strongly correlated to sarcopenia. Methods: We performed a retrospective review of data from patients diagnosed with or without DRF in our institute between 2015 and 2020. Finally, after propensity score matching, data from 115 patients with and 115 patients without DRF were used for analyses. Multivariate logistic regression analysis was performed for sex, body mass index (BMI), the presence of low skeletal muscle mass, bone quality measured by dual-energy X-ray absorptiometry (DXA), and comorbidities (diabetes mellitus, CVD). Results: We found that female sex (odds ratio = 3.435, p = 0.015), CVD (odds ratio = 5.431, p < 0.001) and low skeletal muscle mass (odds ratio = 8.062, p = 0.001) were significant predictors for DRF. BMI and osteoporosis were not statistically significantly related to DRF. Conclusions: Women with low skeletal muscle mass and CVD may be more responsible for DRF than osteoporosis.
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BACKGROUND: One in five patients with mechanical alignment (MA) after total knee arthroplasty (TKA) was reportedly dissatisfied. As constitutional varus knees are common, restoring the patients' natural residual varus (RV) alignment is as an appealing alternative to neutral MA. This meta-analysis aimed to evaluate the effects of RV alignment on the functional outcomes compared with those of MA in TKA for the knees with varus osteoarthritis. METHODS: The MEDLINE/PubMed, Cochrane Library, and EMBASE databases were comprehensively searched for papers comparing the effects of RV alignment and MA on the functional outcomes from the time of inception of the databases to July 2020. Studies comparing the functional outcomes in the knees subjected to TKA with RV alignment (case group) and MA (control group) were included. The Knee Society knee and functional scores (KSKS and KSFS, respectively), Western Ontario and McMaster University Osteoarthritis Index (WOMAC), Oxford knee score (OKS), and forgotten joint score (FJS) were compared. RESULTS: Seven studies were finally included; all studies showed a low risk of selection bias and provided detailed demographic data. The pooled mean difference in the KSKS (0.06, 95% confidence interval [CI]: -0.14 to 0.27; p = 0.55) and KSFS (0.08, 95% CI: -0.08 to 0.35; p = 0.56) between RV alignment and MA did not significantly differ. The pooled mean differences in the WOMAC (-0.25, 95% CI: -0.57 to 0.07; p = 0.12), OKS (0.06, 95% CI: -0.15 to 0.27; p = 0.56), and FJS (0.41, 95% CI: -0.18 to 1.00; p = 0.18) between the groups were not significant. CONCLUSION: The beneficial effects of RV alignment on the functional outcomes are limited compared to those of MA in TKA for varus osteoarthritis to date. Currently, TKA with neutral MA should be considered as the gold standard.
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Artroplastia do Joelho , Prótese do Joelho , Osteoartrite do Joelho , Humanos , Joelho/cirurgia , Articulação do Joelho/cirurgia , Osteoartrite do Joelho/cirurgiaRESUMO
Alzheimer's disease (AD) is a neurodegenerative disease causing progressive cognitive decline until eventual death. AD affects millions of individuals worldwide in the absence of effective treatment options, and its clinical causes are still uncertain. The onset of dementia symptoms indicates severe neurodegeneration has already taken place. Therefore, AD diagnosis at an early stage is essential as it results in more effective therapy to slow its progression. The current clinical diagnosis of AD relies on mental examinations and brain imaging to determine whether patients meet diagnostic criteria, and biomedical research focuses on finding associated biomarkers by using neuroimaging techniques. Multiple clinical brain imaging modalities emerged as potential techniques to study AD, showing a range of capacity in their preciseness to identify the disease. This review presents the advantages and limitations of brain imaging modalities for AD diagnosis and discusses their clinical value.
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Doença de Alzheimer , Disfunção Cognitiva , Doenças Neurodegenerativas , Doença de Alzheimer/diagnóstico por imagem , Doença de Alzheimer/psicologia , Biomarcadores , Encéfalo/diagnóstico por imagem , Disfunção Cognitiva/diagnóstico , Progressão da Doença , Humanos , Neuroimagem/métodosRESUMO
Roll-to-roll nanoimprint lithography (RTR-NIL) is a low-cost and continuous fabrication process for large-area functional films. However, the partial ultraviolet (UV) resin filling obstructs the ongoing production process. This study incorporates UV resin filling process into the nanopillars and nanopores by using RTR-NIL. A multiphase numerical model with a sliding mesh method is proposed in this study to show the actual phenomena of imprint mold rotation and feeding of UV resin on the polyethylene terephthalate (PET) substrate. The implementation of UV resin filling under environmental conditions was performed by utilizing the open-channel (OC) boundary conditions. The numerical model was solved by using the explicit volume of fluid (VOF) scheme to compute the filling on each node of the computational domain. The effects of different processing parameters were investigated through the proposed numerical model such as imprinting speed (IS), contact angles (CAs), viscosity, initial thickness of the PET, and supporting roll diameter. A good agreement was found between numerical simulations and experimental results. The proposed numerical model gives better insights of the filling process for the mass production of functional surfaces with nanopillars and nanopores patterns for different applications on an industrial scale.
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BACKGROUND: This study aimed (1) to introduce a computed tomography (CT)-based classification of the posterolateral compartment of the tibial plateau based on the fibula and to propose the individualized surgical approaches for each zone; and (2) to determine the surgical approach based on the classification, that would achieve a safe and effective reduction and could improve postoperative clinical outcomes. METHODS: Eighteen cases of tibia plateau fracture involving the isolated posterolateral compartment in a single institution were retrospectively analyzed. The posterolateral compartment of the tibial plateau was segmented into three zones based on the fibular position and an individualized surgical approach was proposed for each zone. In anterior Zone I, surgical treatment was performed using an extended anterolateral approach and the patient was placed in the supine position; in middle Zone II, using the transfibular approach in the supine position; in posterior Zone III, using the posteromedial approach in the prone position. RESULTS: In all cases, anatomical articular reduction (intra-articular step off in CT images <2 mm) was achieved and maintained for the follow up period. The average mechanical medial proximal tibial angle was increased from 87.6° before surgery to 88.2° in the immediate postoperative period (P = 0.060), and maintained for the follow up period (mean 89.9° at 1-year follow up). At the 1-year follow up, the knee range of motion averaged 140° and the Lysholm knee function score was 95.0 points. CONCLUSION: An individualized surgical approach and fixation according to three-zone subdivision for isolated posterolateral tibial plateau fractures provided an effective and safe method to treat posterolateral tibial plateau fractures.
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Fixação Interna de Fraturas , Fraturas da Tíbia , Consolidação da Fratura , Humanos , Amplitude de Movimento Articular , Estudos Retrospectivos , Fraturas da Tíbia/diagnóstico por imagem , Fraturas da Tíbia/cirurgia , Resultado do TratamentoRESUMO
Vertically oriented femoral neck fractures (FNFs) are known to be especially unstable FNFs, and they have a higher associated risk of failure. The dynamic hip screw (DHS) technique and the cannulated cancellous screw (CCS) technique are the two main fixation techniques used in the treatment of FNFs. However, no large clinical study has compared the DHS and CCS techniques in patients with high-shear-angle FNFs. MEDLINE, Embase, Cochrane Library, and Web of Science were systematically searched for studies that compared the DHS and CCS techniques for the treatment of Pauwels type II or type III FNF. Pooled analysis was performed to identify differences between the DHS and CCS techniques in Pauwels type II or type III FNF, with a focus on postoperative complications such as fracture nonunion and osteonecrosis of the femoral head (ONFH). We included five studies with a total of 252 patients. The DHS technique was used in 96 patients (DHS group), and the CCS technique was used in 156 patients (CCS group). The pooled analysis revealed that the nonunion rate in the CCS group was significantly higher than that in the DHS group (OR = 0.32; 95% CI, 0.11-0.96; p = 0.04, I2 = 0%), but there was no difference in the incidence of ONFH between the groups (OR = 0.98; 95% CI, 0.20-4.73; p = 0.98, I2 = 53%). For vertically oriented FNFs, the DHS technique is more favorable and has a lower risk of fracture nonunion than the CCS technique.
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The safety of food contact plastic materials, including PP, PE, PET, PCT, PLA, PBT and cross-linked polyester, was assessed with regard to migrated substances. The migrated concentrations of overall migrants (OMs), terephthalic acid, acetaldehyde, 1,4-butanediol and lead, were determined according to the standards and specifications for utensils, containers and packages in Korea. Food simulants of 4% acetic acid, water and n-heptane were used for the analysis of the substances. The dietary exposures of terephthalic acid, acetaldehyde and 1,4-butanediol were assessed using the dietary concentrations and the food consumption data. As a result, the dietary exposures were considered to be safe comparing to the health-based guidance values. In the case of lead, the margin of exposure (MOE) approach was applied. The MOEs calculated using the UB concentration and mean consumption data were ranged from 3 to 1000, which indicated low concern for health risk. Moreover, in this study, the dietary exposures were estimated by the Korean MFDS and U.S. FDA methods, respectively. As a result, the assessed risks were considered to be low in both cases. Based on the results of current exposure assessments, it could be considered that the food contact plastic materials are properly controlled by the regulatory authorities.
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Acetaldeído/análise , Butileno Glicóis/análise , Exposição Dietética , Contaminação de Alimentos/análise , Chumbo/análise , Ácidos Ftálicos/análise , Ácido Acético/química , Embalagem de Alimentos/instrumentação , Armazenamento de Alimentos/instrumentação , Heptanos/química , Humanos , Plásticos/química , República da Coreia , Água/químicaRESUMO
Si surfaces were texturized with periodically arrayed oblique nanopillars using slanted plasma etching, and their optical reflectance was measured. The weighted mean reflectance (Rw) of the nanopillar-arrayed Si substrate decreased monotonically with increasing angles of the nanopillars. This may have resulted from the increase in the aspect ratio of the trenches between the nanopillars at oblique angles due to the shadowing effect. When the aspect ratios of the trenches between the nanopillars at 0° (vertical) and 40° (oblique) were equal, the Rw of the Si substrates arrayed with nanopillars at 40° was lower than that at 0°. This study suggests that surface texturing of Si with oblique nanopillars reduces light reflection compared to using a conventional array of vertical nanopillars.
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Urban development pattern significantly impacts stream water quality by influencing pollutant generation, build-up, and wash-off processes. It is thus necessary to understand and predict stream water quality in accordance with different urban development patterns to effectively advise urban growth planning and policies. To do so, we collected pollutant concentration data on nitrate (NO3--N), total phosphate (TP), and Escherichia coli (E. coli) from 1047 sampling stations in the Texas Gulf Region. We utilized a Random Forest (RF) machine learning model to predict stream water quality under four planning scenarios with different urban densities and configurations. SHapley Additive exPlanations (SHAP) was used to prove the importance of urban development pattern in influencing stream water quality. The spatial variations of the impact of these patterns were explored with Geographically Weighted Regression (GWR). SHAP results indicated that Largest Patch Index (LPI), Patch Cohesion Index (COHESION), Splitting Index (SPLIT), and Landscape Division Index (DIVISION) were the most important urban development pattern metrics affecting stream water quality. The spatial variations of such patterns were shown to impact stream water quality depending on pollutants, seasonality, climate, and urbanization level. RF prediction results suggested that high density aggregated development was more effective in reducing TP and NO3--N concentrations than the current sprawl development, but had the potential risk of increasing E. coli pollution in the wet season. The results of this study provide empirical evidence and a potential mechanistic explanation that stream water quality degradation is a consequence of urban sprawl. Lastly, machine learning is a powerful tool for scenario prediction in land use planning to forecast environmental impacts under different urban development pattern scenarios.
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SERS based immunoassays for point-of-care diagnostics are a promising tool to facilitate biomarker detection for early disease diagnosis and disease control. The technique is based on a sandwiched system in which antigen is first captured by a selective plasmonic paper substrate and then labeled by an extrinsic Raman label (ERL), consisting of a 60 nm gold nanoparticle (AuNP) functionalized with a mixed monolayer of detection antibody and 4-nitrobenezenethiol (NBT) as a Raman reporter molecule. Here, we report on the use of AuNP modified filter paper as a novel capture membrane in a vertical flow format. This vertical flow configuration affords reproducible flow of sample and label through the capture substrate to overcome diffusion limited kinetics and significantly reduced assay time. The filter paper was selected due to its affordability and availability, while the embedded AuNPs maximized plasmonic coupling with the ERLs and SERS enhancement. Additionally, the embedded AuNP served as a scaffold to immobilize capture antibody to specifically bind antigen. In this work, a SERS-based rapid vertical flow (SERS-RVF) immunoassay for detection of mouse IgG was developed to establish proof-of-principle. Optimization of assay conditions led to a limit of detection of 3 ng/mL, which is comparable to more traditional formats carried out in multi-well plates, and significantly reduced assay time to less than 2 min. Additionally, IgG was accurately quantified in normal mouse serum to validate the SERS-RVF assay for application to the analysis of biological samples. These results highlight the potential advantages of the SERS-RVF platform for point-of-need testing.
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Ouro , Nanopartículas Metálicas , Animais , Imunoensaio , Camundongos , Sistemas Automatizados de Assistência Junto ao Leito , Análise Espectral RamanRESUMO
This report describes strategies to increase the reactive surfaces of integrated gold nanoparticles (AuNPs) by employing two different types of host materials that do not possess strong electrostatic and/or covalent interactive forces. These composite particles are then utilized as highly reactive and recyclable quasi-homogeneous catalysts in a C-C bond forming reaction. The use of mesoporous TiO2 and poly(N-isopropylacrylamide), PNIPAM, particles allows for the formation of relatively small and large guest AuNPs and provides the greatly improved stability of the resulting composite particles. As these AuNPs are physically incorporated into the mesoporous TiO2 (i.e., supported AuNPs) and PNIPAM particles (i.e., encapsulated AuNPs), their surfaces are maximized to serve as highly reactive catalytic sites. Given their increased physicochemical properties (e.g., stability, dispersity, and surface area), these composite particles exhibit notably high catalytic activity, selectivity, and recyclability in the homocoupling of phenylboronic acid in water and EtOH. Although the small supported AuNPs display slightly faster reaction rates than the large encapsulated AuNPs, the apparent activation energies (Ea) of both composite particles are comparable, implying no obvious correlation with the size of guest AuNPs under the reaction conditions. Investigating the overall physical properties of various composite particles and their catalytic functions, including the reactivity, selectivity, and Ea, can lead to the development of highly practical quasi-homogeneous catalysts in green reaction conditions.
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Exfoliated graphene oxide (GO) was reliably modified with a cetyltrimethylammonium chloride (CTAC) surfactant to greatly improve the dispersity of the GO in a polyacrylonitrile (PAN) polymer precursor solution. Subsequent electrospinning of the mixture readily resulted in the formation of GO-PAN composite nanofibers containing up to 30 wt % of GO as a filler without notable defects. The absence of common electrospinning problems associated with clogging and phase separation indicated the systematic and uniform integration of the GO within the PAN nanofibers beyond the typical limits. After thoroughly examining the formation and maximum loading efficiency of the modified GO in the PAN nanofibers, the resulting composite nanofibers were thermally treated to form membrane-type sheets. The wettability and pore properties of the composite membranes were notably improved with respect to the pristine PAN nanofiber membrane, possibly due to the reinforcing filler effect. In addition, the more GO loaded into the PAN nanofiber membranes, the higher the removal ability of the methylene blue (MB) and methyl red (MR) dyes in the aqueous system. The adsorption kinetics of a mixed dye solution were also monitored to understand how these MB and MR dyes interact differently with the composite nanofiber membranes. The simple surface modification of the fillers greatly facilitated the integration efficiency and improved the ability to control the overall physical properties of the nanofiber-based membranes, which highly impacted the removal performance of various dyes from water.
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In the field of micro-nanofluidics, a freestanding configuration of a nanoporous junction is highly demanded to increase the design flexibility of the microscale device and the interfacial area between the nanoporous junction and microchannels, thereby improving the functionality and performance. This work first reports direct fabrication and incorporation of a freestanding nanoporous junction in a microfluidic device by performing an electrolyte-assisted electrospinning process to fabricate a freestanding nanofiber membrane and subsequently impregnating the nanofiber membrane with a nanoporous precursor material followed by a solidification process. This process also enables to readily control the geometry of the nanoporous junction depending on its application. By these advantages, vertically stacked 3D micro-nanofluidic devices with complex configurations are easily achieved. To demonstrate the broad applicability of this process in various research fields, a reverse electrodialysis-based energy harvester and an ion concentration polarization-based preconcentrator are produced. The freestanding Nafion-polyvinylidene fluoride nanofiber membrane (F-NPNM) energy harvester generates a high power (59.87 nW) owing to the enlarged interfacial area. Besides, 3D multiplexed and multi-stacked F-NPNM preconcentrators accumulate multiple preconcentrated plugs that can increase the operating sample volume and the degree of freedom of handling. Hence, the proposed process is expected to contribute to numerous research fields related to micro-nanofluidics in the future.