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1.
Anal Biochem ; 408(2): 342-4, 2011 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-20723536

RESUMO

Selective messenger RNA (mRNA) profiling of transfected breast cancer gene expression in a living cell is demonstrated. Atomic force microscope (AFM) probe tips are structurally modified to create a dielectrophoretic force that attracts mRNA molecules within the cell nucleus. The tip end is chemically modified to hybridize only to the target mRNA from a pool of molecules within the nucleus. We successfully combined this scheme with standard assay techniques to develop an assay technology that can be used for early disease detection and basic studies in cell biology.


Assuntos
Neoplasias da Mama/genética , Microscopia de Força Atômica/métodos , RNA Mensageiro/análise , Animais , Linhagem Celular Tumoral , Feminino , Regulação da Expressão Gênica , Gliceraldeído-3-Fosfato Desidrogenases/genética , Microscopia de Força Atômica/instrumentação , Hibridização de Ácido Nucleico/métodos , RNA Mensageiro/metabolismo , Ratos , Receptores Acoplados a Proteínas G/genética , Transfecção
2.
Appl Phys Lett ; 95(8): 83117, 2009 Aug 24.
Artigo em Inglês | MEDLINE | ID: mdl-19777090

RESUMO

We report on a selective and nondestructive measurement of mRNA (messenger ribonucleic acid) expression levels within a living cell. We first modify an atomic force microscope tip to create a tapered nanoscale coaxial cable. Application of an ac (alternating potential) between the inner and outer electrodes of this cable creates a dielectrophoretic force attracting mRNA molecules toward the tip-end which is pretreated with gene specific primers. We selectively extracted and analyzed both high ( approximately 2500) and extremely low (11 0) copy number mRNA from a living cell mRNA in less than 10 s.

3.
Phys Rev E Stat Nonlin Soft Matter Phys ; 73(5 Pt 1): 051914, 2006 May.
Artigo em Inglês | MEDLINE | ID: mdl-16802974

RESUMO

We report on harmonic generation by budding yeast cells (Saccharomyces cerevisiae, 10(8) cells/ml) in response to sinusoidal electric fields with amplitudes ranging from zero to 5 V/cm in the frequency range 10-300 Hz. The cell-generated harmonics are found to exhibit strong amplitude and frequency dependence. Sodium metavanadate, an inhibitor of the proton pump known as H+-ATPase, and glucose, a substrate of H+-ATPase, are found to increase harmonic production at low amplitudes while reducing it at large amplitudes. This P-type proton pump can be driven by an oscillatory transmembrane potential, and its nonlinear response is believed to be largely responsible for harmonic production at low frequencies in yeast cells. We find that the observed harmonics show dramatic changes with time and in their field and frequency dependence after perturbing the system by adding an inhibitor, substrate, or membrane depolarizer to the cell suspension.


Assuntos
Relógios Biológicos/fisiologia , Membrana Celular/fisiologia , Potenciais da Membrana/fisiologia , Modelos Biológicos , Bombas de Próton/fisiologia , Proteínas de Saccharomyces cerevisiae/metabolismo , Saccharomyces cerevisiae/fisiologia , Relógios Biológicos/efeitos da radiação , Membrana Celular/efeitos da radiação , Simulação por Computador , Relação Dose-Resposta à Radiação , Campos Eletromagnéticos , Ativação do Canal Iônico/fisiologia , Ativação do Canal Iônico/efeitos da radiação , Potenciais da Membrana/efeitos da radiação , Proteínas de Membrana/metabolismo , Bombas de Próton/efeitos da radiação , Doses de Radiação , Saccharomyces cerevisiae/efeitos da radiação
4.
Phys Rev Lett ; 95(15): 158103, 2005 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-16241766

RESUMO

We report on harmonic generation by budding yeast cells in response to a sinusoidal electric field, which is seen to be minimal when the field amplitude is less than a threshold value. Surprisingly, sodium metavanadate, an inhibitor of P-type ATPases reportedly responsible for nonlinear response in yeast, reduces the threshold field amplitude, increasing harmonic generation at low amplitudes while reducing it at large amplitudes, whereas the addition of glucose dramatically increases the production of even harmonics. Finally, a simple model is proposed to interpret the observed behavior.


Assuntos
Adenosina Trifosfatases/fisiologia , Adenosina Trifosfatases/efeitos da radiação , Membrana Celular/fisiologia , Membrana Celular/efeitos da radiação , Ativação do Canal Iônico/fisiologia , Ativação do Canal Iônico/efeitos da radiação , Modelos Biológicos , Simulação por Computador , Relação Dose-Resposta à Radiação , Campos Eletromagnéticos , Conformação Proteica/efeitos da radiação , ATPases Translocadoras de Prótons/fisiologia , ATPases Translocadoras de Prótons/efeitos da radiação , Doses de Radiação , Saccharomyces cerevisiae/fisiologia , Saccharomyces cerevisiae/efeitos da radiação
5.
J Chem Phys ; 121(24): 12428-30, 2004 Dec 22.
Artigo em Inglês | MEDLINE | ID: mdl-15606263

RESUMO

Reactions between metals and chloride solutions have been shown to exhibit magnetic field fluctuations over a wide range of size and time scales. Power law behavior observed in these reactions is consistent with models said to exhibit self-organized criticality. Voltage fluctuations observed during the dissolution of magnesium and aluminum in copper chloride solution are qualitatively similar to the recorded magnetic signals. In this paper, distributions of voltage and magnetic peak sizes, noise spectra, and return times are compared for both reactions studied.

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