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1.
Anal Biochem ; 558: 12-18, 2018 10 01.
Artigo em Inglês | MEDLINE | ID: mdl-30076789

RESUMO

Here, we aimed to use graphene oxide to improve the selectivity and sensitivity of Tyr determination via the reaction with 1-nitroso-2-naphthol as a selective reagent of Tyr. The reaction between Tyr and 1-nitroso-2-naphthol in absence and presence of GO was studied spectrophotometrically. Different parameters such as concentrations, temperature, incubation time were optimized. The obtained data showed that the maximum absorbance was achieved by using 2 mL of 0.03% 1-nitroso-2-naphthol at temperature 60 °C for 10 min. On the basis of calibration curve of various concentrations of Tyr in the presence of 20 µg mL-1 GO, the limit of detection was 6.4 × 10-6 M (1.15 µg mL-1), where in absence of GO was 1.1 × 10-5 M (19.9 µg mL-1). The selectivity of Tyr in presence of other amino acids and phenols was studied with and without GO. The data obtained revealed that the selectivity of Tyr in presence of GO with respect to some amino acids and phenols was improved. The proposed method has been applied for the determination of Tyr in urine and serum samples. Therefore, GO is a powerful catalytic surface for the sensitive and selective determination of Try in biological fluids.


Assuntos
Grafite/química , Nanoestruturas/química , Tirosina/análise , Aminoácidos/química , Líquidos Corporais/química , Humanos , Limite de Detecção , Microscopia Eletrônica de Varredura , Fenóis/química , Espectroscopia de Infravermelho com Transformada de Fourier , Tirosina/sangue , Tirosina/urina
2.
Materials (Basel) ; 11(1)2018 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-29300306

RESUMO

Graphene oxide (GO) is emerging as a promising nanomaterial with potential application in the detection and analysis of amino acids, DNA, enzymes, and proteins in biological fluid samples. So, the reaction of GO with amino acids should be characterized and determined before using it in biosensing methods and devices. In this study, the reaction of tyrosine amino acid (Tyr) with GO was characterized using FT-IR, UV-vis spectrophotometry, and scanning electron microscopy (SEM) before its use. The optimum conditions for GO's interaction with Tyr amino acid have been studied under variable conditions. The optimum conditions of pH, temperature, shaking time, and GO and tyrosine concentrations for the uptaking of tyrosine amino acid onto the GO's surface from aqueous solution were determined. The SEM analysis showed that the GO supplied was in a particle size range between 5.4 and 8.1 nm. A pH of 8.4-9.4 at 25 °C and 5 min of shaking time were the optimum conditions for a maximum uptake of 1.4 µg/mL of tyrosine amino acid onto 0.2 mg/mL of GO.

3.
Nanoscale ; 8(37): 16662-16669, 2016 Sep 22.
Artigo em Inglês | MEDLINE | ID: mdl-27714111

RESUMO

Among various cancers, pediatric brain tumors represent the most common cancer type in children and the second most common cause of cancer related deaths. Anticancer drugs and therapies, such as doxorubicin (Dox), have severe side effects on patients during chemotherapy, especially for children as their bodies are still under development. These side effects are believed to be due to the lack of a delivery system with high efficacy and targeting selectivity, resulting in serious damages of normal cells. To improve the efficacy and selectivity, the transferrin (Trans) receptor mediated endocytosis can be utilized for drug delivery system design, as transferrin receptors are expressed on the blood brain barrier (BBB) and often over expressed in brain tumor cells. Carbon dots (C-Dots) have recently emerged as benign nanoparticles in biomedical applications owing to their good water solubility, tunable surface functionalities and excellent biocompatibility. The unique characteristics of C-Dots make them promising candidates for drug delivery development. In this study, carbon dots-transferrin-doxorubicin covalent conjugate (C-Dots-Trans-Dox) was synthesized, characterized by different spectroscopic techniques and investigated for the potential application as a drug delivery system for anticancer drug doxorubicin to treat pediatric brain tumors. Our in vitro results demonstrate greater uptake of the C-Dots-Trans-Dox conjugate compared to Dox alone presumably owing to the high levels of transferrin receptors on these tumor cells. Experiment showed that C-Dots-Trans-Dox at 10 nM was significantly more cytotoxic than Dox alone, reducing viability by 14-45%, across multiple pediatric brain tumor cell lines.


Assuntos
Carbono , Doxorrubicina/administração & dosagem , Sistemas de Liberação de Medicamentos , Nanopartículas , Transferrina , Neoplasias Encefálicas/tratamento farmacológico , Linhagem Celular Tumoral , Humanos , Nanoconjugados
4.
Colloids Surf B Biointerfaces ; 145: 251-256, 2016 Sep 01.
Artigo em Inglês | MEDLINE | ID: mdl-27187189

RESUMO

Drug delivery to the central nervous system (CNS) in biological systems remains a major medical challenge due to the tight junctions between endothelial cells known as the blood-brain-barrier (BBB). Here we use a zebrafish model to explore the possibility of using transferrin-conjugated carbon dots (C-Dots) to ferry compounds across the BBB. C-Dots have previously been reported to inhibit protein fibrillation, and they are also used to deliver drugs for disease treatment. In terms of the potential medical application of C-Dots for the treatment of CNS diseases, one of the most formidable challenges is how to deliver them inside the CNS. To achieve this in this study, human transferrin was covalently conjugated to C-Dots. The conjugates were then injected into the vasculature of zebrafish to examine the possibility of crossing the BBB in vivo via transferrin receptor-mediated endocytosis. The experimental observations suggest that the transferrin-C-Dots can enter the CNS while C-Dots alone cannot.


Assuntos
Barreira Hematoencefálica/metabolismo , Carbono/química , Transferrina/metabolismo , Peixe-Zebra/metabolismo , Animais , Luz , Microscopia Confocal , Modelos Animais
5.
Anal Chim Acta ; 787: 226-32, 2013 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-23830443

RESUMO

Reaction of formaldehyde with amino acids followed by oxidation with hydrogen peroxide to produce a fluorophore Norharman product is well known and was used for the spectrofluorimetric determination of l-tryptophan (Trp). This study aimed to use graphene oxide (GO) to enhance the selectivity and sensitivity of Trp in presence of other amino acids and possible interfering compounds. Different parameters such as pH, temperature, incubation time, and concentrations of formaldehyde, H2O2 and GO were studied to optimize the condition of determination. Experimental data showed that the maximum fluorescence intensity was achieved in pH 7.0-9.0 phosphate buffer mixed with 7-10% (v/v) formaldehyde and 1-2% (v/v) H2O2 as oxidizing agent at 60°C for 1h. On the basis of calibration curve of various concentrations of Trp in the presence of 20 µg mL(-1) GO, the lower limit of detection (LOD) of Trp was determined as 0.092 nmol mL(-1) and the lower limit of quantification (LOQ) was 0.3 nmol mL(-1). The selectivity of Trp in presence of other amino acids and possible interfering compounds were studied with and without GO. The data obtained after inner filter effect corrections revealed that the selectivity of Trp in presence of amino acids and other possible interfering agents was improved in the range of 76-96%, compared with that in absence of GO. The enhancement of selectivity in the presence of GO indicates that the Trp and other amino acid and possible interfering compounds were adsorbed by GO, and the selective uptaking of Trp-by the reaction with formaldehyde followed by oxidation with H2O2 at 60°C with high selectivity and sensitivity was achieved successfully.


Assuntos
Grafite/química , Nanopartículas/química , Óxidos/química , Triptofano/análise , Nanopartículas/normas , Espectrometria de Fluorescência/métodos , Espectrometria de Fluorescência/normas
6.
Anal Bioanal Chem ; 400(3): 787-95, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21360032

RESUMO

The activity of the α-L-fucosidase (AFU) enzyme represents an excellent test for diagnosis of hepatocellular carcinoma (HCC) and fucosidosis recognized in inborn disorder of metabolism and increases the sensitivity of detection to 95.5% in patients with HCC. Therefore, the determination of the activity of AFU enzyme is very important and can be used as a screening tool for the early diagnosis of tumors for HCC patients. A simple, accurate, and sensitive potentiometric method was developed for measuring the activity of AFU. The method was based upon measuring the concentration of 2-chloro-4-nitrophenol (2-chloro-4-NP) using a 2-chloro-4-NP-rhodamine B ion pair in a PVC membrane sensor. The electrode shows a linear, reproducible, and stable potentiometric response with an anionic Nernstian slope of -51.13 ± 0.6 mV/decade over a wide range of concentrations 10(-5)-10(-2) M and a detection limit of 1.0 × 10(-6) M of 2-chloro-4-NP. The membrane exhibits a fast response time of 30 s, over a pH range of 4.0-6.5. The selectivity coefficients indicate excellent selectivity for 2-chloro-4-NP over a number of interfering species, e.g., chloride, nitrate, sulfate, chromate urea, albumin, glucose, uric acid, and total protein. The prepared sensor has been used successfully for the determination of 2-chloro-4-NP produced from the hydrolysis of 2-chloro-4-NP-α-L-fucopyranoside substrate. It was also applied for the determination α-L-fucosidase enzyme of 33 serum samples of healthy subjects and patients. The average recoveries ± RSD for the healthy subjects, cirrhosis of chronic hepatitis C and B, and HCC serum samples were 102.6 ± 1.01%, 101.5 ± 0.95%, and 100.1 ± 1.1%, respectively. The results obtained are in good agreement with those obtained by standard methods.


Assuntos
Técnicas Biossensoriais/métodos , Carcinoma Hepatocelular/enzimologia , Membranas Artificiais , Nitrofenóis/química , Cloreto de Polivinila/química , Rodaminas/química , alfa-L-Fucosidase/sangue , Adulto , Idoso , Técnicas Biossensoriais/economia , Feminino , Hepatite C Crônica/enzimologia , Humanos , Limite de Detecção , Masculino , Pessoa de Meia-Idade , Nitrofenóis/metabolismo , Cloreto de Polivinila/metabolismo , Potenciometria/economia , Potenciometria/métodos , Adulto Jovem , alfa-L-Fucosidase/metabolismo
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