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1.
Anal Chem ; 90(6): 3766-3774, 2018 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-29498256

RESUMO

We describe a technology to perform sizing and concentration analysis of double stranded DNA with a sensitivity of 10 fg/µL in an operating time of 20 min. The technology is operated automatically on a commercial capillary electrophoresis instrument using electro-hydrodynamic actuation. It relies on a new capillary device that achieves online concentration of DNA at the junction between two capillaries of different diameters, thanks to viscoelastic lift forces. Using a set of DNA ladders in the range of 100-1500 bp, we report a sizing accuracy and precision better than 3% and a concentration quantification precision of ∼20%. When the technology is applied to the analysis of clinical samples of circulating cell-free DNA (cfDNA), the measured cfDNA concentrations are in good correlation with those measured by digital PCR. Furthermore, the cfDNA size profiles indicate that the fraction of low molecular weight cfDNA in the range of 75-240 bp is a candidate biomarker to discriminate between healthy subjects and cancer patients. We conclude that our technology is efficient in analyzing highly diluted DNA samples and suggest that it will be helpful in translational and clinical research involving cfDNA.


Assuntos
Ácidos Nucleicos Livres/sangue , Eletroforese Capilar/instrumentação , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/sangue , Ácidos Nucleicos Livres/análise , Desenho de Equipamento , Humanos , Hidrodinâmica , Limite de Detecção , Neoplasias/sangue , Neoplasias/diagnóstico , Reação em Cadeia da Polimerase
2.
Electrophoresis ; 33(12): 1709-14, 2012 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-22740458

RESUMO

LEDs present an alternative to lasers in LIF detection with CE, resulting in LED-induced fluorescence (LEDIF). LEDs are much less expensive, consume less energy and are more stable. In addition, LED light sources allow a greater range of wavelengths to better match the maximum wavelength for the fluorescence of the dye. Antibodies were largely studied in SDS capillary gel electrophoresis (SDS-CGE) and LIF detection with different dyes to label the proteins. In this work, our goal is to show that LEDs can advantageously replace lasers. We used 5-carboxytetramethylrhodamine succinimidyl ester (5-TAMRA.SE), 3-(2-furoyl)-quinoline-2 carboxaldehyde (FQ), and naphthalene-2,3-dialdehyde (NDA) to label IgG and we compared the LIF sensitivity with that obtained from LEDIF. We measured that the LOD values of LEDIF are identical to that obtained with the wavelength equivalent laser, and for 5-TAMRA.SE analysis, LOD values are about six times better than when the classical 488 nm laser was used.


Assuntos
Eletroforese Capilar/instrumentação , Eletroforese Capilar/métodos , Corantes Fluorescentes/química , Imunoglobulina G/análise , Dodecilsulfato de Sódio/química , Espectrometria de Fluorescência/métodos , Aminas/química , Aminoácidos/química , Fluoresceína-5-Isotiocianato , Furanos/química , Humanos , Imunoglobulina G/química , Lasers , Limite de Detecção , Naftalenos/química , Quinolinas/química , Rodaminas/química
3.
Environ Sci Pollut Res Int ; 25(14): 14117-14137, 2018 May.
Artigo em Inglês | MEDLINE | ID: mdl-29520553

RESUMO

Faced with health, environmental, and socio-economic issues related to the heavy use of pesticides, diffuse phytosanitary pollution becomes a major concern shared by all the field actors. These actors, namely the farmers and territorial managers, have expressed the need to implement decision support tools for the territorial management of diffuse pollution resulting from the plant protection practices and their impacts. To meet these steadily increasing requests, a cartographic analysis approach was implemented based on GIS which allows the spatialization of the diffuse pollution impacts related to plant protection practices on the Etang de l'Or catchment area in the South of France. Risk mapping represents a support-decision tool that enables the different field actors to identify and locate vulnerable areas, so as to determine action plans and agri-environmental measures depending on the context of the natural environment. This work shows that mapping is helpful for managing risks related to the use of pesticides in agriculture by employing indicators of pressure (TFI) and risk on the applicator's health (IRSA) and on the environment (IRTE). These indicators were designed to assess the impact of plant protection practices at various spatial scales (field, farm, etc.). The cartographic analysis of risks related to plant protection practices shows that diffuse pollution is unequally located in the North (known for its abundant garrigues and vineyards) and in the South of the Etang de l'Or catchment area (the Mauguio-Lunel agricultural plain known for its diversified cropping systems). This spatial inequity is essentially related to land use and agricultural production system. Indeed, the agricultural lands cover about 60% of the total catchment area. Consequently, this cartographic analysis helps the territorial actors with the implementation of strategies for managing risks of diffuse pollution related to pesticides use in agriculture, based on environmental and socio-economic issues and the characteristics of the natural environment.


Assuntos
Agricultura/métodos , Monitoramento Ambiental/métodos , Poluição Ambiental/análise , Poluição Ambiental/prevenção & controle , Praguicidas/análise , Gestão de Riscos/métodos , Área Programática de Saúde , França , Análise Espacial
4.
Environ Sci Pollut Res Int ; 24(8): 6951-6972, 2017 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-27164887

RESUMO

The health, environmental and socio-economic issues related to the massive use of plant protection products are a concern for all the stakeholders involved in the agricultural sector. These stakeholders, including farmers and territorial actors, have expressed a need for decision-support tools for the management of diffuse pollution related to plant protection practices and their impacts. To meet the needs expressed by the public authorities and the territorial actors for such decision-support tools, we have developed a technical-economic model "OptiPhy" for risk mitigation based on indicators of pesticide toxicity risk to applicator health (IRSA) and to the environment (IRTE), under the constraint of suitable economic outcomes. This technical-economic optimisation model is based on linear programming techniques and offers various scenarios to help the different actors in choosing plant protection products, depending on their different levels of constraints and aspirations. The health and environmental risk indicators can be broken down into sub-indicators so that management can be tailored to the context. This model for technical-economic optimisation and management of plant protection practices can analyse scenarios for the reduction of pesticide-related risks by proposing combinations of substitution PPPs, according to criteria of efficiency, economic performance and vulnerability of the natural environment. The results of the scenarios obtained on real ITKs in different cropping systems show that it is possible to reduce the PPP pressure (TFI) and reduce toxicity risks to applicator health (IRSA) and to the environment (IRTE) by up to approximately 50 %.


Assuntos
Produção Agrícola/métodos , Produtos Agrícolas/crescimento & desenvolvimento , Técnicas de Apoio para a Decisão , Modelos Teóricos , Praguicidas/toxicidade , Gestão de Riscos/métodos , Produção Agrícola/economia , Produtos Agrícolas/efeitos dos fármacos , Humanos , Modelos Econômicos , Praguicidas/análise , Gestão de Riscos/economia
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