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1.
Materials (Basel) ; 16(20)2023 Oct 12.
Artigo em Inglês | MEDLINE | ID: mdl-37895647

RESUMO

The cubic nonlinearity of a graphene-oxide monolayer was characterized through open and closed z-scan experiments, using a nano-second laser operating at a 10 Hz repetition rate and featuring a Gaussian spatial beam profile. The open z-scan revealed a reverse saturable absorption, indicating a positive nonlinear absorption coefficient, while the closed z-scan displayed valley-peak traces, indicative of positive nonlinear refraction. This observation suggests that, under the given excitation wavelength, a two-photon or two-step excitation process occurs due to the increased absorption in both the lower visible and upper UV wavelength regions. This finding implies that graphene oxide exhibits a higher excited-state absorption cross-section compared to its ground state. The resulting nonlinear absorption and nonlinear refraction coefficients were estimated to be approximately ~2.62 × 10-8 m/W and 3.9 × 10-15 m2/W, respectively. Additionally, this study sheds light on the interplay between nonlinear absorption and nonlinear refraction traces, providing valuable insights into the material's optical properties.

2.
Int J Mol Sci ; 23(9)2022 Apr 26.
Artigo em Inglês | MEDLINE | ID: mdl-35563168

RESUMO

Muscle atrophy is a major muscle disease, the symptoms of which include decreased muscle volume leading to insufficient muscular support during exercise. One cause of muscle atrophy is the induction of oxidative stress by reactive oxygen species (ROS). This study aimed to identify the antioxidant mechanism of linoleic acid (LA) in muscle atrophy caused by oxidative stress. H2O2 has been used to induce oxidative stress in myoblasts in vitro. C2C12 myoblasts treated with H2O2 exhibited decreased viability and increased ROS synthesis. However, with LA treatment, the cells tended to recover from oxidative effects similar to those of the control groups. At the molecular level, the expression of superoxide dismutase 1 (SOD1), Bax, heat shock protein 70 (HSP70), and phosphorylated forkhead box protein O1 was increased by oxidative stress, causing apoptosis. LA treatment suppressed these changes. In addition, the expression of MuRF1 and Atrogin-1/MAFbx mRNA increased under oxidative stress but not in the LA-treated group. Sciatic denervation of C57BL/6 mice manifested as atrophy of the skeletal muscle in micro-computed tomography (micro-CT). The protein expression levels of SOD1, HSP70, and MuRF1 did not differ between the atrophied muscle tissues and C2C12 myoblasts under oxidative stress. With LA treatment, muscle atrophy recovered and protein expression was restored to levels similar to those in the control. Therefore, this study suggests that LA may be a candidate substance for preventing muscle atrophy.


Assuntos
Peróxido de Hidrogênio , Ácido Linoleico , Animais , Denervação , Peróxido de Hidrogênio/metabolismo , Ácido Linoleico/metabolismo , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/metabolismo , Atrofia Muscular/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Superóxido Dismutase-1/metabolismo , Microtomografia por Raio-X
3.
Sensors (Basel) ; 18(1)2017 Dec 26.
Artigo em Inglês | MEDLINE | ID: mdl-29278402

RESUMO

Using gold nanoparticles (AuNPs) on "capillary enzyme-linked immunosorbent assay (ELISA)", we produced highly sensitive and rapid assays, which are the major attributes for point-of-care applications. First, in order to understand the size effect of AuNPs, AuNPs of varying diameters (5 nm, 10 nm, 15 nm, 20 nm, 30 nm, and 50 nm) conjugated with Horseradish Peroxidase (HRP)-labeled anti-C reactive protein (antiCRP) (AuNP•antiCRP-HRP) were used for well-plate ELISA. AuNP of 10 nm produced the largest optical density, enabling detection of 0.1 ng/mL of CRP with only 30 s of incubation, in contrast to 10 ng/mL for the ELISA run in the absence of AuNP. Then, AuNP of 10 nm conjugated with antiCRP-HRP (AuNP•antiCRP-HRP) was used for "capillary ELISA" to detect as low as 0.1 ng/mL of CRP. Also, kinetic study on both 96-well plates and in a capillary tube using antiCRP-HRP or AuNP•antiCRP-HRP showed a synergistic effect between AuNP and the capillary system, in which the fastest assay was observed from the "AuNP capillary ELISA", with its maximum absorbance reaching 2.5 min, while the slowest was the typical well-plate ELISA with its maximum absorbance reaching in 13.5 min.


Assuntos
Ensaio de Imunoadsorção Enzimática , Anticorpos , Ouro , Peroxidase do Rábano Silvestre , Nanopartículas Metálicas
4.
ACS Appl Mater Interfaces ; 9(50): 43490-43497, 2017 Dec 20.
Artigo em Inglês | MEDLINE | ID: mdl-29171259

RESUMO

Molybdenum disulfide (MoS2) field-effect transistor (FET)-based biosensors have attracted significant attention as promising candidates for highly sensitive, label-free biomolecule detection devices. In this paper, toward practical applications of biosensors, we demonstrate reliable and quantitative detection of a prostate cancer biomarker using the MoS2 FET biosensor in a nonaqueous environment by reducing nonspecific molecular binding events and realizing uniform chemisorption of anti-PSA onto the MoS2 surface. A systematic and statistical study on the capability of the proposed device is presented, and the biological binding events are directly confirmed and characterized through intensive structural and electrical analysis. Our proposed biosensor can reliably detect various PSA concentrations with a limit of 100 fg/mL. Moreover, rigorous theoretical simulations provide a comprehensive understanding of the operating mechanism of the MoS2 FET biosensors, and further suggests the enhancement of the sensitivity through engineering device design parameters.


Assuntos
Técnicas Biossensoriais , Desenho de Equipamento , Humanos , Masculino , Sistemas Automatizados de Assistência Junto ao Leito , Neoplasias da Próstata
5.
J Nanosci Nanotechnol ; 16(2): 1942-4, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-27433706

RESUMO

Plasmon-coupled CdSe/ZnS and CdTe/CdS/ZnS coreshells are investigated for their optoelectronic applications because of their high color purity, wide optical tunability, large PL enhancement, and compact and easy integration into electronic devices. The quantum confinement of carriers within quantum dots (QDs) with sizes near the exciton Bohr radius (CdSe ~ 5.8 nm, CdTe ~ 7 nm) exhibits the features of discrete energy states and blue-shift from the bulk bandgap (CdSe ~718 nm, CdTe ~ 863 nm) in the optical spectrum. While the fluorescence from the QDs is attributable to the exciton carrier recombination, large PL enhancement and fast emission time is achieved through plasmon-exciton coupling via the Coulomb interaction. Large PL enhancement of QDs in the vicinity of plasmonic particles was observed and attributed to the reduction of the non-radiative decay rate and large local field enhancement. The large PL enhancement and wide optical tunability along with high color purity from plasmon-coupled QDs enables the realization of hybrid LEDs.

6.
Biosci Biotechnol Biochem ; 80(5): 911-9, 2016 May.
Artigo em Inglês | MEDLINE | ID: mdl-26943486

RESUMO

Pterocarpus indicus Willd has been widely used as a traditional medicine to treat edema, cancer, and hyperlipidemia, but its antiallergic properties and underlying mechanisms have not yet been studied. The purpose of this study was to evaluate the antiallergic activity of Pterocarpus indicus Willd water extract (PIW) using activated mast cells and an atopic dermatitis (AD)-like mouse model. PIW decreased IgE/Ag-induced mast cell degranulation and the phosphorylation of Syk and downstream signaling molecules such as PLC-γ, Akt, Erk 1/2, JNK compared to stimulated mast cells. In DNCB-induced AD-like mice, PIW reduced IgE level in serum, as well as AD-associated scratching behavior and skin severity score. These results indicate that PIW inhibits the allergic response by reducing mast cell activation and may have clinical potential as an antiallergic agent for disorders such as AD.


Assuntos
Antialérgicos/farmacologia , Dermatite Atópica/tratamento farmacológico , Imunoglobulina E/sangue , Mastócitos/efeitos dos fármacos , Pterocarpus/química , Pele/efeitos dos fármacos , Animais , Degranulação Celular/efeitos dos fármacos , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/imunologia , Dermatite Atópica/patologia , Dinitroclorobenzeno , Modelos Animais de Doenças , Regulação da Expressão Gênica , Irritantes , MAP Quinase Quinase 4/genética , MAP Quinase Quinase 4/imunologia , Masculino , Mastócitos/imunologia , Mastócitos/patologia , Camundongos , Camundongos Endogâmicos ICR , Proteína Quinase 1 Ativada por Mitógeno/genética , Proteína Quinase 1 Ativada por Mitógeno/imunologia , Proteína Quinase 3 Ativada por Mitógeno/genética , Proteína Quinase 3 Ativada por Mitógeno/imunologia , Fosfolipase C gama/genética , Fosfolipase C gama/imunologia , Fosforilação/efeitos dos fármacos , Extratos Vegetais/química , Proteínas Proto-Oncogênicas c-akt/genética , Proteínas Proto-Oncogênicas c-akt/imunologia , Transdução de Sinais , Pele/imunologia , Pele/patologia , Quinase Syk/genética , Quinase Syk/imunologia , Água
7.
Biosci Biotechnol Biochem ; 80(6): 1095-106, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26924430

RESUMO

Angiogenesis, neovascularization from pre-existing vessels, is a key step in tumor growth and metastasis, and anti-angiogenic agents that can interfere with these essential steps of cancer development are a promising strategy for human cancer treatment. In this study, we characterized the anti-angiogenic effects of Coptis japonica Makino extract (CJME) and its mechanism of action. CJME significantly inhibited the proliferation, migration, and invasion of vascular endothelial growth factor (VEGF)-stimulated HUVECs. Furthermore, CJME suppressed VEGF-induced tube formation in vitro and VEGF-induced microvessel sprouting ex vivo. According to our study, CJME blocked VEGF-induced cell cycle transition in G1. CJME decreased expression of cell cycle-regulated proteins, including Cyclin D, Cyclin E, Cdk2, and Cdk4 in response to VEGF. Taken together, the results of our study indicate that CJME suppresses VEGF-induced angiogenic events such as proliferation, migration, and tube formation via cell cycle arrest in G1.


Assuntos
Inibidores da Angiogênese/farmacologia , Coptis/química , Pontos de Checagem da Fase G1 do Ciclo Celular/efeitos dos fármacos , Regulação da Expressão Gênica/efeitos dos fármacos , Neovascularização Patológica/prevenção & controle , Fator A de Crescimento do Endotélio Vascular/antagonistas & inibidores , Inibidores da Angiogênese/isolamento & purificação , Animais , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Ciclina D/antagonistas & inibidores , Ciclina D/genética , Ciclina D/metabolismo , Ciclina E/antagonistas & inibidores , Ciclina E/genética , Ciclina E/metabolismo , Quinase 2 Dependente de Ciclina/antagonistas & inibidores , Quinase 2 Dependente de Ciclina/genética , Quinase 2 Dependente de Ciclina/metabolismo , Quinase 4 Dependente de Ciclina/antagonistas & inibidores , Quinase 4 Dependente de Ciclina/genética , Quinase 4 Dependente de Ciclina/metabolismo , Pontos de Checagem da Fase G1 do Ciclo Celular/genética , Células Endoteliais da Veia Umbilical Humana , Humanos , Masculino , Neovascularização Patológica/genética , Neovascularização Patológica/metabolismo , Extratos Vegetais/química , Ratos , Ratos Sprague-Dawley , Fator A de Crescimento do Endotélio Vascular/farmacologia
8.
Artigo em Inglês | MEDLINE | ID: mdl-26981139

RESUMO

Cymbidium has known antibacterial and antiedema activity and has been used as an ingredient in cosmetics and fragrances. The effects of Cymbidium ethanol extract (CYM) on allergic response and the underlying mechanisms of action have not been reported. Therefore, the purpose of this study was to determine the effect of CYM on allergic responses. Topical application of CYM was effective against immunoglobulin E (IgE)/dinitrophenyl-conjugated bovine serum albumin- (DNP-BSA-) induced degranulation of RBL-2H3 cells and anaphylaxis in ICR mice. An allergic dermatitis-like mouse model was used to evaluate the therapeutic potential of CYM in vivo. Continuous application of 2,4-dinitrochlorobenzene (DNCB) not only induced dermatitis in ICR mice but also aggravated the skin lesioning. However, the application of CYM decreased skin lesion severity, scratching behavior, and IgE levels. In addition, CYM downregulated the expression of the proinflammatory cytokines interleukin- (IL-) 4, IL-13, and tumor necrosis factor- (TNF-) α. Studies of signal transduction pathways showed that CYM suppressed the phosphorylation of spleen tyrosine kinase (Syk), an upstream molecule. It also inhibited the phosphorylation of Akt, phospholipase C- (PLC-) γ, and mitogen-activated protein kinase kinase kinase (MEKK). These results indicate that CYM may be effective in preventing and reducing allergic response and may have therapeutic potential as an antiallergic agent in disorders such as atopic dermatitis.

9.
Biosci Biotechnol Biochem ; 80(1): 80-8, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26613402

RESUMO

Skeletal muscle atrophy can be defined as a decrease of muscle volume caused by injury or lack of use. This condition is associated with reactive oxygen species (ROS), resulting in various muscular disorders. We acquired 2D and 3D images using micro-computed tomography in gastrocnemius and soleus muscles of sciatic-denervated mice. We confirmed that sciatic denervation-small animal model reduced muscle volume. However, the intraperitoneal injection of Oenothera odorata root extract (EVP) delayed muscle atrophy compared to a control group. We also investigated the mechanism of muscle atrophy's relationship with ROS. EVP suppressed expression of SOD1, and increased expression of HSP70, in both H2O2-treated C2C12 myoblasts and sciatic-denervated mice. Moreover, EVP regulated apoptotic signals, including caspase-3, Bax, Bcl-2, and ceramide. These results indicate that EVP has a positive effect on reducing the effect of ROS on muscle atrophy.


Assuntos
Músculo Esquelético/efeitos dos fármacos , Atrofia Muscular/tratamento farmacológico , Oenothera/química , Extratos Vegetais/farmacologia , Espécies Reativas de Oxigênio/antagonistas & inibidores , Transdução de Sinais/efeitos dos fármacos , Animais , Apoptose/efeitos dos fármacos , Caspase 3/genética , Caspase 3/metabolismo , Linhagem Celular , Ceramidas/metabolismo , Denervação/efeitos adversos , Modelos Animais de Doenças , Regulação da Expressão Gênica , Proteínas de Choque Térmico HSP70/agonistas , Proteínas de Choque Térmico HSP70/genética , Proteínas de Choque Térmico HSP70/metabolismo , Injeções Intraperitoneais , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Músculo Esquelético/metabolismo , Músculo Esquelético/patologia , Atrofia Muscular/etiologia , Atrofia Muscular/metabolismo , Atrofia Muscular/patologia , Mioblastos/citologia , Mioblastos/efeitos dos fármacos , Mioblastos/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Espécies Reativas de Oxigênio/metabolismo , Nervo Isquiático/cirurgia , Superóxido Dismutase/antagonistas & inibidores , Superóxido Dismutase/genética , Superóxido Dismutase/metabolismo , Superóxido Dismutase-1 , Proteína X Associada a bcl-2/genética , Proteína X Associada a bcl-2/metabolismo
10.
Eur J Pharmacol ; 744: 36-41, 2014 Dec 05.
Artigo em Inglês | MEDLINE | ID: mdl-25261039

RESUMO

Abnormal proliferation of vascular smooth muscle cells (VSMCs) plays an essential functional role in the pathogenesis of vascular disorders, such as atherosclerosis, restenosis, and neointimal hyperplasia. In this study, we examined the effects of meso-dihydroguaiaretic acid (MDGA) on platelet-derived growth factor (PDGF)-BB-induced proliferation and the molecular basis of its underlying mechanism of action in rat aortic VSMCs. Incubation of resting VSMCs with MDGA for 24 h significantly diminished PDGF-BB-induced DNA synthesis in a dose-dependent manner. We also examined the effects of MDGA on PDGF-BB signal transduction. Pre-treatment of VSMCs with MDGA inhibited PDGF-BB-induced phosphorylation of extracellular signal-regulated kinases 1/2 (ERK1/2), p38, and C-Jun N-terminal kinase (JNK). MDGA also effectively inhibited phosphorylation of Akt, phospholipase C gamma 1 (PLCγ1), and PDGF receptor beta (PDGFRß). These results indicate that MDGA may inhibit proliferation of VSMCs by suppressing autophosphorylation of PDGFRß, and may be useful in the treatment of VSMC-associated vascular disease such as atherosclerosis, restenosis, and neointimal hyperplasia after angioplasty.


Assuntos
Aorta/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Guaiacol/análogos & derivados , Lignanas/farmacologia , Músculo Liso Vascular/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Fosforilação/efeitos dos fármacos , Proteínas Proto-Oncogênicas c-sis/metabolismo , Animais , Aorta/metabolismo , Becaplermina , Células Cultivadas , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Guaiacol/farmacologia , Proteínas Quinases JNK Ativadas por Mitógeno/metabolismo , Músculo Liso Vascular/metabolismo , Miócitos de Músculo Liso/metabolismo , Fosfolipase C gama/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Ratos , Transdução de Sinais/efeitos dos fármacos , Proteínas Quinases p38 Ativadas por Mitógeno/metabolismo
11.
Biosci Biotechnol Biochem ; 78(9): 1568-71, 2014.
Artigo em Inglês | MEDLINE | ID: mdl-25209505

RESUMO

Nodakenin, derived from the roots of Angelica gigas Nakai, is an important natural resource and medicinal material with anti-allergic and anti- inflammatory activities. We have previously shown that nodakenin inhibits IgE/Ag-induced degranulation in mast cells. In this study, we investigated the inhibitory effect of nodakenin on 2,4-dinitrochlorobenzene (DNCB)-induced atopic dermatitis (AD)- like skin lesions in ICR mice. Scratching behavior, skin severity score, blood IgE level, and skin thickness were improved in DNCB-induced AD-like ICR mice. Our results showed that nodakenin suppressed the increase of AD-like skin lesions in ICR mice. These results suggest that nodakenin may be a potential therapeutic resource for AD as well as an adjunctive agent to control itching associated with AD.


Assuntos
Cumarínicos/administração & dosagem , Dermatite Atópica/tratamento farmacológico , Glucosídeos/administração & dosagem , Prurido/tratamento farmacológico , Pele/efeitos dos fármacos , Angelica/química , Animais , Degranulação Celular/efeitos dos fármacos , Dermatite Atópica/induzido quimicamente , Dermatite Atópica/metabolismo , Humanos , Camundongos , Nitrobenzoatos/toxicidade , Prurido/induzido quimicamente , Prurido/patologia , Pele/patologia
12.
Adv Mater ; 25(40): 5701-5, 2013 Oct 25.
Artigo em Inglês | MEDLINE | ID: mdl-23946273

RESUMO

Conductive, flexible, and durable reduced RGO textiles with a facile preparation method are presented. BSA proteins serve as universal adhesives for improving the adsorption of GO onto any textile, irrespective of the materials and the surface conditions. Using this method, we successfully prepared various RGO textiles based on nylon-6 yarns, cotton yarns, polyester yarns, and nonwoven fabrics.


Assuntos
Grafite/química , Soroalbumina Bovina/química , Adsorção , Animais , Caprolactama/análogos & derivados , Caprolactama/química , Bovinos , Óxidos/química , Poliésteres/química , Polímeros/química , Soroalbumina Bovina/metabolismo , Eletricidade Estática
13.
Lab Chip ; 12(18): 3272-6, 2012 Sep 21.
Artigo em Inglês | MEDLINE | ID: mdl-22832792

RESUMO

We present a novel blood filter unit that is designed to separate blood plasma from whole blood by simple magnetic actuation. A non-diluted blood sample is dropped into the filter unit and magnetic attraction is applied to squeeze out only blood plasma while blood particles are filtered by membranes stacked in the filter unit. The new filter device yields good filtering performance with nearly perfect filtering efficiency (∼99.999%), high plasma recovery (∼30%), low blood consumption (<50 µl), and fast operation (∼1 min). Because it is simple to operate and is attachable to any kind of pre-made biochip, it has commercial potential in various lab-on-a-chip applications for blood tests.


Assuntos
Filtração/instrumentação , Magnetismo , Plasma/química , Desenho de Equipamento , Humanos , Técnicas Analíticas Microfluídicas/instrumentação , Técnicas Analíticas Microfluídicas/métodos
14.
Biosens Bioelectron ; 33(1): 233-40, 2012 Mar 15.
Artigo em Inglês | MEDLINE | ID: mdl-22305674

RESUMO

This paper describes a new technique for the detection of uncharged or feebly charged small molecules (<400Da) using Si field-effect transistor (FET) biosensors that are signal-enhanced by gold nanoparticle (NP) charges under dry measurement conditions. NP charges are quickly induced by a chemical deposition (that is, Au deposition) and the indirect competitive immunogold assay, and strongly enhance the electrical signals of the FET biosensors. For the validation of signal enhancement of FET biosensors based on NP charges and detection of uncharged or feebly charged small molecules, mycotoxins (MTXs) of aflatoxin-B1 (AFB1), zearalenone (ZEN), and ochratoxin-A (OTA) were used as target molecules. According to our experimental results, the signal is 100 times more enhanced than the use of the existing solution FET biosensing techniques. Furthermore, this method enables the FET biosensor to quantitatively detect target molecules, regardless of the ionic strengths, isoelectric points (pI), or pHs of the measured sample solutions.


Assuntos
Aflatoxina B1/análise , Técnicas Biossensoriais/instrumentação , Ocratoxinas/análise , Transistores Eletrônicos , Zearalenona/análise , Ouro/química , Nanopartículas Metálicas/química
15.
Methods Mol Biol ; 854: 67-85, 2012.
Artigo em Inglês | MEDLINE | ID: mdl-22311754

RESUMO

The application of difference gel electrophoresis (DIGE), in particular its most common "minimal labeling" variety, utilizes N-hydroxysuccinimide esters of Cy2, Cy3, and Cy5 dyes, which are commercially available. We describe methods for the efficient synthesis of all three dyes from relatively inexpensive and commercially available precursors in only a few steps and with relatively high yields. In model DIGE experiments, the newly synthesized dyes proved to be indistinguishable from commercially available ones and have been shown to be stable for years while stored under argon as dry solids or after being dissolved in N,N-dimethylformamide.


Assuntos
Carbocianinas/síntese química , Técnicas de Química Sintética/métodos , Corantes/síntese química , Eletroforese em Gel Diferencial Bidimensional/métodos , Carbocianinas/química , Corantes/química , Estabilidade de Medicamentos , Armazenamento de Medicamentos , Focalização Isoelétrica , Reprodutibilidade dos Testes , Fatores de Tempo
16.
Opt Mater Express ; 2(8): 1026-1039, 2012 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-23457661

RESUMO

A hybrid optical nanostructure of plasmon-coupled SQDs was developed for photonic applications. The coupling distances between the mono-layers of Au nanoparticles with a surface concentration of ~9.18 × 10-4 nm-2 and CdSe/ZnS SQDs with that of ~3.7 × 10-3 nm-2 were controlled by PMMA plasma etching. Time-resolved spectroscopy of plasmon-coupled SQDs revealed a strong shortening of the longest lifetime and ~9-fold PL enhancement. Polarization-resolved PL spectroscopy displayed linear polarization and depolarization at near- and far-field plasmon-coupling, respectively. The physical origin of PL enhancement could be attributable to both the large local field enhancement and the fast resonant energy transfer.

17.
Biosens Bioelectron ; 29(1): 219-23, 2011 Nov 15.
Artigo em Inglês | MEDLINE | ID: mdl-21890339

RESUMO

We propose a new type of photosensitive biosensor with a CMOS compatible Si photodiode integrated circuit, for the high-sensitive detection of small mycotoxin molecules requiring competitive assay approach. In this work, a photodiode is connected to the gate of a field effect transistor (FET) so that the open circuit voltage (V(OC)) of the illuminated photodiode is transferred into the drain/source current (I(DS)) of the FET. The sensing scheme employs competitive binding of toxin molecules (within the sample solution) and toxin-BSA conjugates (immobilized on the photodiode surface) with Au-nanoparticle-labeled antibodies, followed by silver enhancement to generate opaque structures on the photodiode surface. By utilizing the non-linear dependence of the V(OC) on the light intensity, we can maintain a sufficiently high signal resolution at low toxin concentrations (with most of the incident light blocked) for the competitive assay. By monitoring the I(DS) of the FET whose gate is driven by the V(OC), quantitative detection of Aflatoxin B1 has been achieved in the range of 0-15ppb.


Assuntos
Aflatoxina B1/análise , Técnicas Biossensoriais/métodos , Aflatoxina B1/imunologia , Anticorpos Imobilizados , Anticorpos Monoclonais , Técnicas Biossensoriais/estatística & dados numéricos , Técnicas Eletroquímicas , Ouro , Imunoensaio/métodos , Imunoensaio/estatística & dados numéricos , Nanopartículas Metálicas/ultraestrutura , Microscopia Eletrônica de Varredura , Processos Fotoquímicos , Silício
18.
Anal Chem ; 82(23): 9686-93, 2010 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-21049960

RESUMO

Cardiac markers in human serum with concentrations less than 0.1 ng/mL were analyzed by use of a guided-mode resonance (GMR) biosensor. Cardiac troponin I (cTnI), creatine kinase MB (CK-MB), and myoglobin (MYO) were monitored in the serum of both patients and healthy controls. Dose-response curves ranging from 0.05 to 10 ng/mL for cTnI, from 0.1 to 10 ng/mL for CK-MB, and from 0.03 to 1.7 µg/mL for MYO were obtained. The limits of detection (LOD) for cTnI, CK-MB, and MYO were less than 0.05, 0.1, and 35 ng/mL, respectively. Analysis time was 30 min, which is short enough to meet clinical requirements. Antibody immobilization and the hydrophilic properties of the guided-mode resonance filter (GMRF) surface were investigated by X-ray photoelectron spectroscopy (XPS) and by monitoring the peak wavelength shift and water contact angle (CA). Both assays used to evaluate the surface density of the immobilized antibodies, a sandwich enzyme-linked immunosorbent assay (ELISA) and a sandwich immunogold assay, showed that the antibodies were successfully immobilized and sufficiently aligned to detect the low concentration of biomarkers. Our results show that the GMR biosensor will be very useful in developing low-cost portable biosensors that can screen for cardiac diseases.


Assuntos
Técnicas Biossensoriais/métodos , Creatina Quinase Forma MB/sangue , Mioglobina/sangue , Troponina I/sangue , Anticorpos Imobilizados/química , Anticorpos Imobilizados/imunologia , Biomarcadores/sangue , Ouro/química , Humanos , Espectroscopia Fotoeletrônica
19.
Langmuir ; 26(10): 7355-64, 2010 May 18.
Artigo em Inglês | MEDLINE | ID: mdl-20205399

RESUMO

Anionic sulfate (SO(4)(-))-functionalized polystyrene (PS) nanoparticles were prepared by the thermal decomposition of potassium persulfate (KPS) in the presence of sodium tetraborate via emulsion polymerization. The presence of a SO(4)(-) group at a solid/liquid interface of a particle surface was confirmed by a zeta potential value of -40.6 mV as well as the shifting of S 2p spectra toward a lower-binding-energy region around 162.7 eV (2p(3/2)) and 164.4 eV (2p(1/2)) in X-ray photoelectron spectroscopy (XPS) analysis. The electrostatic attraction between positively charged antibodies of human immunoglobulin G (hIgG) and cardiac troponin I (cTnI) and negatively charged particle surfaces was accomplished. The atomic force microscopy (AFM) measurement and bicinchoninic acid (BCA) assay results show binding structure between hIgG and antibodies of hIgG (anti-hIgG) with a gradual increase in particle diameter to 152.6 nm (bare), 170.2 nm (hIgG), and 178.9 nm (hIgG/anti-hIgG). Surface coverage densities of 331.4 ng/cm(2) (hIgG) and 320.3 ng/cm(2) (cTnI) and the binding capacity of hIgG to HyLite-750-labeled Fab-specific anti-hIgG (approximately 81.2%) indicate that the majority of hIgG was immobilized with a Y-shaped orientation. The sandwich immunoassay results provide the evidence that the immunological activity of cTnI on the PS nanoparticle surface was retained because the binding activity of the cTnI-PS nanoparticle/cTnI (antigen)/detection cTnI-antibody reaction showed a 5-fold higher activity than that of the cTnI-PS nanoparticle/human serum albumin (HSA)/detection cTnI antibody used as a negative control.


Assuntos
Imunoensaio , Imunoglobulina G/química , Nanopartículas/química , Poliestirenos/química , Sulfatos/química , Troponina I/química , Ânions/química , Anticorpos/química , Anticorpos/imunologia , Reações Antígeno-Anticorpo , Humanos , Imunoglobulina G/imunologia , Modelos Imunológicos , Tamanho da Partícula , Poliestirenos/síntese química , Poliestirenos/imunologia , Sulfatos/síntese química , Sulfatos/imunologia , Propriedades de Superfície , Troponina I/imunologia
20.
Langmuir ; 25(19): 11692-7, 2009 Oct 06.
Artigo em Inglês | MEDLINE | ID: mdl-19788222

RESUMO

An antibody immobilization was investigated using a self-assembled monolayer (SAM) over the highly refractive coatings with a SiO2, TiO2, or Si3N4 substrate. The immobilization was characterized by analyzing the hydrophilic properties of hydroxyl (OH) groups on surface coatings with contact angle (CA) measurements to enhance protein immobilization. The hydroxyl (OH) group was formed in greater amounts as the oxygen plasma exposure time was increased, which resulted in a large enhancement in antibody immobilization. It indicated that hydroxyl (OH) group formation is critical for developing a label-free optical transducer with a high sensitivity.


Assuntos
Anticorpos Imobilizados/química , Hidróxidos/química , Titânio/química , Aminas/química , Animais , Anticorpos Imobilizados/análise , Anticorpos Imobilizados/imunologia , Anticorpos Monoclonais/análise , Anticorpos Monoclonais/química , Anticorpos Monoclonais/imunologia , Bovinos , Ensaio de Imunoadsorção Enzimática , Interações Hidrofóbicas e Hidrofílicas , Nanopartículas Metálicas/química , Oxigênio/química , Antígeno Prostático Específico/imunologia , Propriedades de Superfície , Molhabilidade
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