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1.
Protein Pept Lett ; 17(9): 1148-54, 2010 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-20394580

RESUMO

A Surface Plasmon Resonance Imaging (SPRI) sensor has been developed for highly selective determination of cathepsin D (Cat D) or/and E (Cat E). The sensor contains immobilised pepstatin A, which binds aspartyl proteases from solution. Pepstatin A activated with N-Hydroxysuccinimide (NHS) and N-Ethyl-N'-(3-dimethylaminopropyl) carbodiimide (EDC) was immobilized on an amine-modified gold surface. Cysteamine was used for modification of the gold surface. Pepstatin A concentration and pH of interaction were optimised. A concentration of pepstatin equal to 0.5 microg mL(-1) and a pH of 3.75 were selected as optimal. The sensor's dynamic response range is between 0.25 and 1.0 ng mL(-1), and the detection limit is 0.12 ng mL(-1). However, the sensor cannot distinguish between Cat D and Cat E. In order to demonstrate the sensor's potential, Cat E was determined in human red blood cells, Cat D in human saliva, as well as total concentration of Cat D and Cat E in human nasal polyps.


Assuntos
Técnicas Biossensoriais/métodos , Catepsinas/análise , Ressonância de Plasmônio de Superfície/métodos , Animais , Catepsina D/análise , Catepsina E/análise , Eritrócitos/química , Humanos , Pólipos Nasais/química , Pepstatinas/análise , Saliva/química
2.
Oral Microbiol Immunol ; 18(6): 405-7, 2003 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-14622349

RESUMO

The in vitro secreted aspartyl proteinase (SAP) activity of Candida albicans isolated from a variety of oral conditions, including healthy oral cavities, was determined. SAP activity (units/10(6) cells/ml, +/-SD) was 0.28 +/- 0.33 for pseudomembranous candidosis isolates (n = 18), 0.35 +/- 0.46 for chronic erythematous candidosis isolates (n = 21) and 0.30 +/- 0.32 for chronic hyperplastic candidosis isolates (n = 50). SAP activity of 0.19 +/- 0.22 was recorded for isolates from squamous cell carcinoma (n = 18), 0.26 +/- 0.37 for burning mouth syndrome isolates (n = 29), 0.25 +/- 0.38 for isolates from xerostomia (n = 15) and 0.39 +/- 0.50 for isolates from lichen planus (n = 13). The SAP activity of isolates from oral disease states was significantly (P < 0.05) higher than that recorded for 28 isolates from healthy mouths (activity of 0.04 +/- 0.03). However, there was no significant difference in the SAP activity between the three forms of clinical oral candidosis (P > 0.05). SAP activity was inhibited in control samples containing the SAP inhibitor, pepstatin A. These results indicate that C. albicans strains associated with oral disease have inherently higher SAP activity.


Assuntos
Ácido Aspártico Endopeptidases/metabolismo , Candida albicans/enzimologia , Doenças da Boca/enzimologia , Síndrome da Ardência Bucal/enzimologia , Candidíase Bucal/enzimologia , Carcinoma de Células Escamosas/enzimologia , Doença Crônica , Eritema/microbiologia , Humanos , Hiperplasia , Líquen Plano Bucal/enzimologia , Neoplasias Bucais/enzimologia , Pepstatinas/análise , Inibidores de Proteases/análise , Xerostomia/enzimologia
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