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1.
Arch Pharm Res ; 46(8): 659-678, 2023 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-37592169

RESUMO

Green synthesis strategies have been widely applied for the preparation of versatile nanomaterials. Gold nanospheres with an average size of 6.95 ± 2.25 nm were green synthesized by using a 70% ethanol extract of Korean red ginseng (Panax ginseng Meyer) root as a reducing agent. A seed-mediated synthesis was conducted to prepare Au-Ag bimetallic nanoparticles using gold nanospheres as seeds. Remarkably, Au-Ag bimetallic nanoparticles with an average size of 80.4 ± 11.9 nm were synthesized. Scanning transmission electron microscopy, energy dispersive X-ray spectroscopy and elemental mappings revealed bimetallic nanoparticles with Au-Ag alloy core and Au-rich shells. A face-centered cubic structure of Au-Ag bimetallic nanoparticles was confirmed by X-ray diffraction analysis. For Au-Ag bimetallic nanoparticles, the ratio of Ag/Au was 0.20 which was detected and analyzed by inductively coupled plasma-mass spectrometry. Gold nanospheres and Au-Ag bimetallic nanoparticles were functionalized by PEGylation, folic acid conjugation and grafting onto graphene oxide. Finally, docetaxel was loaded for evaluating the in vitro cell viability on cancer cells. Successful functionalization was confirmed by Fourier-transform infrared spectra. The anticancer activity of the docetaxel-loaded nanoparticles was higher than that of their non-docetaxel-loaded counterparts. The highest anticancer activity on human gastric adenocarcinoma cells (AGS) was observed in the docetaxel-loaded gold nanospheres that were functionalized by PEGylation, folic acid conjugation and grafting onto graphene oxide. Additionally, grafting onto graphene oxide and docetaxel loading induced high intracellular reactive oxygen species generation. For chemo-photothermal (PTT) anticancer therapy, cell viability was investigated using near-infrared laser irradiation at 808 nm. The highest chemo-PTT anticancer activity on AGS cells was observed in the docetaxel-loaded Au-Ag bimetallic nanoparticles. Therefore, the newly prepared docetaxel-loaded Au-Ag bimetallic nanoparticles in the current report have potential applications in chemo-PTT anticancer therapy.

2.
Front Psychiatry ; 14: 1161200, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37426108

RESUMO

Background: The Coronavirus (COVID-19) pandemic resulted in a dramatic increase in the prevalence of anxiety and depression globally. Although the impact on the mental health of young adults was especially strong, its underlying mechanisms remain elusive. Materials and methods: Using a network approach, the present study investigated the putative pathways between pandemic-related factors and anxiety and depressive symptoms among young adults in South Korea and the U.S. Network analyses were conducted on cross-country data collected during the COVID-19 lockdown period (n = 1,036). Our model included depression symptoms (PHQ-9), generalized anxiety symptoms (GAD-7), and COVID-19-related factors (e.g., COVID-19-related traumatic stress, pandemic concerns, access to medical/mental health services). Results: The overall structure of pandemic-to-symptom networks of South Korea and the U.S. were found to be similar. In both countries, COVID-related stress and negative future anticipation (an anxiety symptom) were identified as bridging nodes between pandemic-related factors and psychological distress. In addition, worry-related symptoms (e.g., excessive worry, uncontrollable worry) were identified as key contributors in maintaining the overall pandemic-to-symptom network in both countries. Conclusion: The similar network structures and patterns observed in both countries imply that there may exist a stable relationship between the pandemic and internalizing symptoms above and beyond the sociocultural differences. The current findings provide new insights into the common potential pathway between the pandemic and internalizing symptoms in South Korea and in the U.S. and inform policymakers and mental health professionals of potential intervention targets to alleviate internalizing symptoms.

3.
Analyst ; 148(6): 1349-1361, 2023 Mar 13.
Artigo em Inglês | MEDLINE | ID: mdl-36857647

RESUMO

An electrochemical immunoassay based on the redox cycling method was presented using vertically paired electrodes (VPEs), which were fabricated using poly(3,4-ethylenedioxythiophene):poly(styrenesulfonate) (PEDOT:PSS) as an electrode material and parylene-C as a dielectric layer. For the application to immunoassays, different electrochemical properties of PEDOT:PSS were analyzed for the redox reaction of 3,3',5,5'-tetramethylbenzidine (TMB, the chromogenic substrate for enzyme-immunoassays) at different pH conditions, including the conductivity (σ), electron transfer rate constant (kapp), and double-layer capacitance (Cdl). The influencing factors on the sensitivity of redox cycling based on VPE based on PEDOT:PSS were analyzed for the redox reaction of TMB, such as the electrode gap and number of electrode pairs. Computer simulation was also performed for the redox cycling results based on VPEs, which had limitations in fabrication, such as VPEs with an electrode gap of less than 100 nm and more than five electrode pairs. Finally, the redox cycling based on VPE was applied to the medical diagnosis of human hepatitis-C virus (hHCV) using a commercial ELISA kit. The sensitivity of the redox cycling method for the medical diagnosis of hHCV was compared with conventional assay methods, such as TMB-based chromogenic detection, luminol-based chemiluminescence assay, and a rapid test kit (lateral flow immunoassay).


Assuntos
Simulação por Computador , Humanos , Eletrodos , Oxirredução , Imunoensaio , Técnicas Imunoenzimáticas
4.
Analyst ; 147(16): 3783-3794, 2022 Aug 08.
Artigo em Inglês | MEDLINE | ID: mdl-35876175

RESUMO

In this study, parylene-C films from plasma deposition as well as thermal deposition were pyrolyzed to prepare a carbon electrode for application in electrochemical immunoassays. Plasma deposition could prepare parylene-C in a faster deposition rate and more precise control over the thickness in comparison with the conventional thermal deposition. To analyze the influence of the deposition method, the crystal and electronic structures of the pyrolyzed parylene-C films obtained via both deposition methods were compared using Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy. For application as a carbon electrode in immunoassays, the electrochemical properties of the pyrolyzed carbon films from two both deposition methods were analyzed, including the double layer capacitance (2.10 µF cm-2 for plasma deposition and 2.20 µF cm-2 for thermal deposition), the apparent electron transfer rate (approximately 1.1 × 10-3 cm s-1 for both methods), and the electrochemical window (approximately -1.0 ∼ 2.1 V for both methods). Finally, the applicability of the pyrolyzed carbon electrode from parylene-C was demonstrated for the diagnosis of human hepatitis-C using various amperometric methods, such as cyclic voltammetry, chronoamperometry, square-wave voltammetry and differential pulse voltammetry.


Assuntos
Carbono , Pirólise , Carbono/química , Eletrodos , Humanos , Imunoensaio , Polímeros , Xilenos
5.
Biosens Bioelectron ; 202: 113975, 2022 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-35042131

RESUMO

Vertically paired electrodes (VPEs) with multiple electrode pairs were developed for the enhancement of capacitive measurements by optimizing the electrode gap and number of electrode pairs. The electrode was fabricated using a conductive polymer layer of PEDOT:PSS instead of Ag and Pt metal electrodes to increase the VPE fabrication yield because the PEDOT:PSS layer could be effectively etched using a reactive dry etching process. In this study, sensitivity enhancement was realized by decreasing the electrode gap and increasing the number of VPE electrode pairs. Such an increase in sensitivity according to the electrode gap and the number of electrode pairs was estimated using a model analyte for an immunoassay. Additionally, a computer simulation was performed using VPEs with different electrode gaps and numbers of VPE electrode pairs. Finally, VPEs with multiple electrode pairs were applied for SARS-CoV-2 nucleoprotein (NP) detection. The capacitive biosensor based on the VPE with immobilized anti-SARS-CoV-2 NP was applied for the specific detection of SARS-CoV-2 in viral cultures. Using viral cultures of SARS-CoV-2, SARS-CoV, MERS-CoV, and CoV-strain 229E, the limit of detection (LOD) was estimated to satisfy the cutoff value (dilution factor of 1/800) for the medical diagnosis of COVID-19, and the assay results from the capacitive biosensor were compared with commercial rapid kit based on a lateral flow immunoassay.


Assuntos
Técnicas Biossensoriais , COVID-19 , Técnicas Biossensoriais/métodos , Simulação por Computador , Eletrodos , Humanos , SARS-CoV-2 , Sensibilidade e Especificidade
6.
Front Psychol ; 13: 1025638, 2022.
Artigo em Inglês | MEDLINE | ID: mdl-36591014

RESUMO

This study defined adherence to COVID-19 prevention guidelines as health behavior and examined whether the two constructs of the health belief model (i.e., perceived susceptibility and perceived severity) and sense of control predict the level of adherence among young adults in South Korea. An online survey (N = 200) conducted in June 2021, showed that perceived susceptibility, perceived severity, and sense of control positively predict adherence behavior. Sense of control significantly moderated the relationship between perceived susceptibility and adherence even after controlling for depression and perceived health status. Specifically, individuals with a lower level of perceived susceptibility still adhered to COVID-19 prevention guidelines if they had a higher level of sense of control. The finding demonstrates the key role of sense of control in promoting adherence to COVID-19 prevention guidelines and the relationship between sense of control and two constructs of the health belief model. Implication for public messaging targeted at young adults during pandemic situations is also discussed.

7.
Adv Healthc Mater ; 10(17): e2002280, 2021 09.
Artigo em Inglês | MEDLINE | ID: mdl-33724723

RESUMO

Wearable healthcare devices have garnered substantial interest for the realization of personal health management by monitoring the physiological parameters of individuals. Attaining the integrity between the devices and the biological interfaces is one of the greatest challenges to achieving high-quality body information in dynamic conditions. Liquid metals, which exist in the liquid phase at room temperatures, are advanced intensively as conductors for deformable devices because of their excellent stretchability and self-healing ability. The unique surface chemistry of liquid metals allows the development of various sensors and devices in wearable form. Also, the biocompatibility of liquid metals, which is verified through numerous biomedical applications, holds immense potential in uses on the surface and inside of a living body. Here, the recent progress of liquid metal-based wearable electronic devices for healthcare with respect to the featured properties and the processing technologies is discussed. Representative examples of applications such as biosensors, neural interfaces, and a soft interconnection for devices are reviewed. The current challenges and prospects for further development are also discussed, and the future directions of advances in the latest research are explored.


Assuntos
Técnicas Biossensoriais , Dispositivos Eletrônicos Vestíveis , Atenção à Saúde , Eletrônica , Humanos , Metais
8.
Adv Sci (Weinh) ; 8(4): 2002497, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33643790

RESUMO

Cellular senescence can either support or inhibit cancer progression. Here, it is shown that intratumoral infiltration of CD8+ T cells is negatively associated with the proportion of senescent tumor cells in colorectal cancer (CRC). Gene expression analysis reveals increased expression of C-X-C motif chemokine ligand 12 (CXCL12) and colony stimulating factor 1 (CSF1) in senescent tumor cells. Senescent tumor cells inhibit CD8+ T cell infiltration by secreting a high concentration of CXCL12, which induces a loss of CXCR4 in T cells that result in impaired directional migration. CSF1 from senescent tumor cells enhance monocyte differentiation into M2 macrophages, which inhibit CD8+ T cell activation. Neutralization of CXCL12/CSF1 increases the effect of anti-PD1 antibody in allograft tumors. Furthermore, inhibition of CXCL12 from senescent tumor cells enhances T cell infiltration and results in reducing the number and size of tumors in azoxymethane (AOM)/dextran sulfate sodium (DSS)-induced CRC. These findings suggest senescent tumor cells generate a cytokine barrier protecting nonsenescent tumor cells from immune attack and provide a new target for overcoming the immunotherapy resistance of CRC.

9.
Enzyme Microb Technol ; 144: 109721, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33541568

RESUMO

In this work, we present a novel microbial biosensor for Salmonella based on impedance spectrometry by using isolated antibodies against a specific bacterial strain from human serum. Anti-Salmonella (or BL21(DE3)) antibodies were isolated from human serum using S. enteritidis (or BL21(DE3)) and the mutant strain ClearColi. After the purification steps, the purification yield of the antibodies was calculated to be 0.2 %. From the FACS analysis, the isolated anti-Salmonella antibodies were estimated to have more than 6-fold higher binding affinity for S. enteritidis compared to antibodies against other kinds of Gram-negative bacterial strains, including HB101, ClearColi, JM110, DH5α, and BL21(DE3). Finally, the anti-Salmonella antibodies isolated herein were used for bacterial detection using electrochemical biosensors based on impedance spectrometry and the Rct value of the antibodies was estimated for S. enteritidis from the Nyquist plot. The limit of detection of the isolated anti-Salmonella antibodies was estimated to be 1.0 × 103 cells/mL for S. enteritidis and 1.0 × 106 cells/mL for BL21(DE3), respectively.


Assuntos
Anticorpos Antibacterianos , Técnicas Biossensoriais , Humanos , Salmonella enteritidis
10.
Enzyme Microb Technol ; 144: 109728, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33541571

RESUMO

In this work phospholipid quantification was carried out using an enzymatic assay based on cyclic voltammetry of the condensation product of N-ethyl-N-(2-hydroxy-3-sulfopropyl)-3,5-dimethoxyaniline sodium salt (DAOS) and 4-aminoantipyrine (4-AP) with a graphite electrode. For the optimization of electrochemical measurement for the product, electrochemical properties such as the electrochemical window, double layer capacitance (Cdl) and electron transfer rate (kapp) were analyzed for a graphite-electrode and Au-electrode. The phospholipid enzymatic assay based the on electrochemical measurement using the graphite electrode was applied to the diagnosis of sepsis for sera from healthy volunteers (n = 16), patients with systemic inflammatory response syndrome (SIRS, n = 16) and severe sepsis patients (n = 24). Finally, the phospholipid quantification results from the electrochemical measurement were statistically compared with the conventional method based on optical density measurement.


Assuntos
Grafite , Sepse , Técnicas Eletroquímicas , Eletrodos , Ensaios Enzimáticos , Humanos , Fosfolipídeos , Sepse/diagnóstico
11.
ACS Appl Bio Mater ; 4(11): 7779-7789, 2021 11 15.
Artigo em Inglês | MEDLINE | ID: mdl-35006761

RESUMO

As the shelf life of platelets collected from donated blood is very short, approximately 5 days, the determination of bacterial contamination in platelets has become necessary. In this study, rapid analysis of Gram-positive and Gram-negative bacterial contamination in platelet samples was presented without pre-enrichment using pig serum-derived antibodies against the outer membrane proteins (OMP) of Gram-negative bacteria and antibodies against lipoteichoic acid (LTA) on the surface of Gram-positive bacteria. The anti-OMP antibodies against Gram-negative bacteria were isolated using sequential incubation with (1) the modified Gram-negative bacteria ClearColi, which lacks lipopolysaccharide (LPS) on the outer membrane, and (2) the Gram-positive bacteriaBacillus subtilis to filter away nonspecifically bound proteins from ClearColi. The anti-lipoteichoic acid (LTA) antibodies against Gram-positive bacteria were isolated using sequential incubation with (1) the Gram-positive bacteriaB. subtilis and (2) the Gram-negative bacteria Escherichia coli BL21 to filter away nonspecifically bound proteins fromB. subtilis. The feasibility of using the antibodies isolated from pig serum against Gram-negative and Gram-positive bacteria was demonstrated using flow cytometry. Finally, detection of the contamination of platelets with Gram-negative and Gram-positive bacteria using the impedance immunosensor based on these isolated antibodies was successfully demonstrated.


Assuntos
Técnicas Biossensoriais , Animais , Anticorpos/metabolismo , Plaquetas , Escherichia coli , Bactérias Gram-Negativas/metabolismo , Bactérias Gram-Positivas/metabolismo , Imunoensaio , Proteínas de Membrana/metabolismo , Suínos
12.
Restor Dent Endod ; 45(2): e4, 2020 May.
Artigo em Inglês | MEDLINE | ID: mdl-32483529

RESUMO

Although numerous studies have been conducted on apexification using mineral trioxide aggregate (MTA), direct observation of extracted human teeth after the procedure has been rarely reported. This case report describes a mandibular premolar treated 2.5 years ago and extracted recently for orthodontic treatment. The tubercle of the right mandibular premolar of a 12-year-old boy with dens evaginatus was fractured and the pulp was exposed. The tooth was diagnosed with pulp necrosis and asymptomatic periapical abscess. During the first visit, copious irrigation was performed with 2.5% sodium hypochlorite. Calcium hydroxide paste was placed as an intracanal medicament. The sinus tract had disappeared at the second visit after 3 weeks. MTA was applied on to the bleeding point as a 4-mm-thick layer, followed by a 3-mm-thick gutta-percha filling and resin core build-up. After 2.5 years, the tooth and three other premolars were extracted for orthodontic treatment. The right and left mandibular premolars were scanned with micro-computed tomography to determine the root shape and canal anatomy. Irregular root growth was observed and the root outline of the right mandibular premolar differed from that of the contralateral tooth. Apexification with MTA leads to the formation of roots with irregular morphology, without any pulpal space.

13.
Biosens Bioelectron ; 156: 112110, 2020 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-32174550

RESUMO

Thermophoresis is the physical diffusion of molecules from hot to cold induced by a thermal gradient. Thermophoresis has been used to evaluate the interaction of biomolecules in solution. In this study, the outer membrane from E. coli was isolated and used to produce OM particles with a diameter of approximately 100 nm. These prepared OM particles were applied in a thermophoretic immunoassay. First, outer membrane (OM) particles with lipopolysaccharides (LPS) and anti-LPS antibodies were used as a model to demonstrate proof of concept and the difference in E. coli thermophoresis was explained by the changes in the molecular surface area (A) and effective charge (σeff). The hydrodynamic size of the molecules was measured as a changing parameter, molecular surface area (A), by dynamic laser scattering (DLS), and the zeta potential was measured as a changing parameter of effective charge (σeff) and then evaluated by the Soret equation. Using the hydrodynamic size and zeta potential values, the interaction between the antigen (OM particle with LPS) and antibody (anti-LPS antibodies) could be monitored and the results were fitted to the thermophoretic immunoassay using the Soret coefficient and equation. Finally, this OM-based immunoassay was applied to the medical diagnosis of systemic lupus erythematosus. Here, OM particles with Ro and La proteins were used to analyze the autoantibodies in patient and control sera. Thermophoretic immunoassay results were also compared to the fitted analysis using hydrodynamic size and zeta potential values and the Soret coefficient and equation.


Assuntos
Membrana Externa Bacteriana/química , Técnicas Biossensoriais/métodos , Escherichia coli/química , Lúpus Eritematoso Sistêmico/diagnóstico , Autoanticorpos/imunologia , Humanos , Hidrodinâmica , Imunoensaio/métodos , Lipopolissacarídeos/imunologia , Lúpus Eritematoso Sistêmico/imunologia , Tamanho da Partícula
14.
ACS Appl Mater Interfaces ; 12(2): 2679-2686, 2020 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-31849212

RESUMO

Two-dimensional (2D) metal dichalcogenides have drawn considerable interest because they offer possibilities for the implementation of emerging electronics. The emerging electronics are moving toward two major directions: vertical expansion of device space and flexibility. However, the development of a synthesis method for 2D metal dichalcogenides that meets all the requirements remains a significant challenge. Here, we propose a promising method for wafer-scale, conformal, and low-temperature (≤240 °C) synthesis of single-phase SnS2 via the atomic layer deposition technique. There is a trade-off relationship between the crystallinity and orientation preference of SnS2, which is efficiently eliminated by the two-step growth occurring at different temperatures. Consequently, the van der Waals layers of the highly crystalline SnS2 are parallel to the substrate. Thin-film transistors (TFTs) comprising the SnS2 layer show reasonable electrical performances (field-effect mobility: ∼0.8 cm2 V-1 s-1 and on/off ratio: ∼106), which are comparable to that of a single-crystal SnS2 flake. Moreover, we demonstrate nonplanar and flexible TFTs to identify the feasibility of the implementation of future electronics. Both the diagonal-structured TFT and flexible TFT fabricated without a transfer process show electrical performances comparable to those of rigid and planar TFTs. Particularly, the flexible TFT does not exhibit substantial degradation even after 2000 bending cycles. Our work would provide decisive opportunities for the implementation of future electronic devices utilizing 2D metal chalcogenides.

15.
Anal Chim Acta ; 1073: 30-38, 2019 Sep 27.
Artigo em Inglês | MEDLINE | ID: mdl-31146833

RESUMO

Highly N-doped SiC was presented as an optimal electrode for electrochemical immunoassays with a far higher sensitivity than chemiluminescence detection. As the first step, the electrochemical properties of highly N-doped SiC, such as the double-layer capacitance (Cdl), rate constant for electron transfer (kapp) and ideal polarizable potential range (electrochemical window) were analyzed and compared with those of Au, Pt, and graphite electrodes. The highly N-doped SiC electrode was used for the quantification of oxidized 3,3',5,5'-tetramethylbenzidine (TMB) which was widely used as chromogenic substrate for commercialized immunoassay kits. In order to enhance the sensitivity for the quantification of the oxidized TMB the chronoamperometry was applied to avoid the background current of i-V measurement. Finally, the chronoamperometry based on the highly N-doped SiC electrode was applied to commercial immunoassay kits for the medical diagnosis of the human immunodeficiency virus (HIV) and the human hepatitis B surface antigen (hHBsAg). The chronoamperometric measurement based on the highly N-doped SiC electrode was proved to detect at far lower limits in comparison with the conventional optical density measurement as well as the chemiluminescence assay based on luminol as a chemiluminescent probe.


Assuntos
Benzidinas/análise , Técnicas Biossensoriais , Compostos Inorgânicos de Carbono/química , Técnicas Eletroquímicas , HIV/isolamento & purificação , Antígenos de Superfície da Hepatite B/isolamento & purificação , Imunoensaio , Compostos de Silício/química , Eletrodos , Humanos
16.
ACS Appl Mater Interfaces ; 11(16): 14892-14901, 2019 Apr 24.
Artigo em Inglês | MEDLINE | ID: mdl-30945837

RESUMO

A new deposition technique is required to grow the active oxide semiconductor layer for emerging oxide electronics beyond the conventional sputtering technique. Atomic layer deposition (ALD) has the benefits of versatile composition control, low defect density in films, and conformal growth over a complex structure, which can hardly be obtained with sputtering. This study demonstrates the feasibility of growing amorphous In-Zn-Sn-O (a-IZTO) through ALD for oxide thin-film transistor (TFT) applications. In the ALD of the a-IZTO film, the growth behavior indicates that there exists a growth correlation between the precursor molecules and the film surface where the ALD reaction occurs. This provides a detailed understanding of the ALD process that is required for precise composition control. The a-IZTO film with In/Zn/Sn = 10:70:20 was chosen for high-performance TFTs, among other compositions, regarding the field-effect mobility (µFE), turn-on voltage ( Von), and subthreshold swing (SS) voltage. The optimized TFT device with the a-IZTO film thickness of 8 nm revealed a high performance with a µFE of 22 cm2 V-1 s-1, Von of 0.8 V, and SS of 0.15 V dec-1 after annealing at 400 °C for 30 min. Furthermore, an emerging device such as a vertical channel TFT was demonstrated. Thus, the a-IZTO ALD process could offer promising opportunities for a variety of emerging oxide electronics beyond planar TFTs.

17.
Anal Chim Acta ; 1055: 106-114, 2019 May 09.
Artigo em Inglês | MEDLINE | ID: mdl-30782361

RESUMO

The autoimmune diseases systemic lupus erythematosus (SLE) and Sjogren's syndrome (SS) are diagnosed by detection of autoantibodies against Ro and La protein autoantigens, respectively. In this work, the diagnosis of autoimmune diseases SLE and SS was demonstrated using thermophoresis of Escherichia coli with the autodisplayed autoantigens. Ro and La protein autoantigens were autodisplayed by constitutive expression together with a fluorescent protein called tdTomato in the cytosol. The binding affinity of the autodisplayed autoantigens was tested against positive and control sera by using FACS as a reference method. The factors influencing interactions between E. coli with autodisplayed autoantigens and autoantibodies in sera during thermophoresis were analyzed by measurement of cell surface charge and size before and after interaction. Finally, the thermophoretic diagnosis of autoimmune diseases SLE and SS was demonstrated using sera from patients afflicted with the respective diseases by estimating sensitivity and selectivity from ROC plots.


Assuntos
Autoantígenos/imunologia , Doenças Autoimunes/diagnóstico , Doenças Autoimunes/imunologia , Escherichia coli/metabolismo , Animais , Autoanticorpos/química , Autoanticorpos/imunologia , Autoanticorpos/metabolismo , Autoantígenos/metabolismo , Modelos Moleculares , Conformação Proteica
18.
Sensors (Basel) ; 19(24)2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31888286

RESUMO

An electrochemical capacitance immunosensor based on an interdigitated wave-shaped micro electrode array (IDWµE) for direct and label-free detection of C-reactive protein (CRP) was reported. A self-assembled monolayer (SAM) of dithiobis (succinimidyl propionate) (DTSP) was used to modify the electrode array for antibody immobilization. The SAM functionalized electrode array was characterized morphologically by atomic force microscopy (AFM) and energy dispersive X-ray spectroscopy (EDX). The nature of gold-sulfur interactions on SAM-treated electrode array was probed by X-ray photoelectron spectroscopy (XPS). The covalent linking of anti-CRP-antibodies onto the SAM modified electrode array was characterized morphologically through AFM, and electrochemically through cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The application of phosphate-buffered saline (PBS) and human serum (HS) samples containing different concentrations of CRP in the electrode array caused changes in the electrode interfacial capacitance upon CRP binding. CRP concentrations in PBS and HS were determined quantitatively by measuring the change in capacitance (ΔC) through EIS. The electrode immobilized with anti-CRP-antibodies showed an increase in ΔC with the addition of CRP concentrations over a range of 0.01-10,000 ng mL-1. The electrode showed detection limits of 0.025 ng mL-1 and 0.23 ng mL-1 (S/N = 3) in PBS and HS, respectively. The biosensor showed a good reproducibility (relative standard deviation (RSD), 1.70%), repeatability (RSD, 1.95%), and adequate selectivity in presence of interferents towards CRP detection. The sensor also exhibited a significant storage stability of 2 weeks at 4 °C in 1× PBS.


Assuntos
Proteína C-Reativa/análise , Técnicas Eletroquímicas/métodos , Animais , Anticorpos/metabolismo , Bovinos , Espectroscopia Dielétrica , Capacitância Elétrica , Humanos , Concentração de Íons de Hidrogênio , Microeletrodos , Microscopia de Força Atômica , Espectroscopia Fotoeletrônica , Reprodutibilidade dos Testes , Soroalbumina Bovina/metabolismo , Succinimidas/química
19.
Biosens Bioelectron ; 126: 518-528, 2019 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-30476883

RESUMO

In this work, the gram-negative bacterium Escherichia coli strain BL21(DE3) (with lipopolysaccharide (LPS) in its outer membrane) and its modified ClearColi™ strain (lacking LPS) were used for the separation of anti-LPS antibodies from human serum by the following steps: (1) binding of the serum proteins to BL21(DE3); (2) dissociation of the bound proteins (including anti-LPS antibodies) from BL21(DE3) with acid; (3) filtering of the dissociated proteins using ClearColi to remove unwanted proteins; and (4) separation of the antibody fraction by protein-A column chromatography. The binding properties of the separated antibodies were analyzed by fluorescence-activated cell sorting to confirm their selective binding to LPS on the outer membrane of BL21(DE3), and by thermophoretic immunoassay to estimate their dissociation constant. The in vitro applicability of the separated anti-LPS antibodies was demonstrated through a fluorescence assay of BL21(DE3), after immobilizing the antibodies onto a modified microplate surface. The electrochemical detection of BL21(DE3) was also achieved after immobilizing the anti-LPS antibodies onto a gold electrode.


Assuntos
Anticorpos Anti-Idiotípicos/sangue , Técnicas Biossensoriais , Escherichia coli/isolamento & purificação , Lipopolissacarídeos/isolamento & purificação , Anticorpos Anti-Idiotípicos/química , Humanos , Imunoensaio , Lipopolissacarídeos/sangue
20.
Sensors (Basel) ; 18(11)2018 Nov 12.
Artigo em Inglês | MEDLINE | ID: mdl-30424510

RESUMO

We report the electrical characteristics and pH responses of a Si-nanonet ion-sensitive field-effect transistor with ultra-thin parylene-H as a gate sensing membrane. The fabricated device shows excellent DC characteristics: a low subthreshold swing of 85 mV/dec, a high current on/off ratio of ~107 and a low gate leakage current of ~10-10 A. The low interface trap density of 1.04 × 1012 cm-2 and high field-effect mobility of 510 cm²V-1s-1 were obtained. The pH responses of the devices were evaluated in various pH buffer solutions. A high pH sensitivity of 48.1 ± 0.5 mV/pH with a device-to-device variation of ~6.1% was achieved. From the low-frequency noise characterization, the signal-to-noise ratio was extracted as high as ~3400 A/A with the lowest noise equivalent pH value of ~0.002 pH. These excellent intrinsic electrical and pH sensing performances suggest that parylene-H can be promising as a sensing membrane in an ISFET-based biosensor platform.

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