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1.
Proc Natl Acad Sci U S A ; 120(37): e2305995120, 2023 09 12.
Artigo em Inglês | MEDLINE | ID: mdl-37669392

RESUMO

To minimize the incorrect use of antibiotics, there is a great need for rapid and inexpensive tests to identify the pathogens that cause an infection. The gold standard of pathogen identification is based on the recognition of DNA sequences that are unique for a given pathogen. Here, we propose and test a strategy to develop simple, fast, and highly sensitive biosensors that make use of multivalency. Our approach uses DNA-functionalized polystyrene colloids that distinguish pathogens on the basis of the frequency of selected short DNA sequences in their genome. Importantly, our method uses entire genomes and does not require nucleic acid amplification. Polystyrene colloids grafted with specially designed surface DNA probes can bind cooperatively to frequently repeated sequences along the entire genome of the target bacteria, resulting in the formation of large and easily detectable colloidal aggregates. Our detection strategy allows "mix and read" detection of the target analyte; it is robust and highly sensitive over a wide concentration range covering, in the case of our test target genome Escherichia coli bl21-de3, 10 orders of magnitude from [Formula: see text] to [Formula: see text] copies/mL. The sensitivity compares well with state-of-the-art sensing techniques and has excellent specificity against nontarget bacteria. When applied to real samples, the proposed technique shows an excellent recovery rate. Our detection strategy opens the way to developing a robust platform for pathogen detection in the fields of food safety, disease control, and environmental monitoring.


Assuntos
DNA , Poliestirenos , Antibacterianos , Coloides , Monitoramento Ambiental , Escherichia coli
2.
Langmuir ; 27(13): 8351-8, 2011 Jul 05.
Artigo em Inglês | MEDLINE | ID: mdl-21650180

RESUMO

Natural lung surfactant contains less than 40% disaturated phospholipids, mainly dipalmitoylphosphatidylcholine (DPPC). The mechanism by which lung surfactant achieves very low near-zero surface tensions, well below its equilibrium value, is not fully understood. To date, the low surface tension of lung surfactant is usually explained by a squeeze-out model which predicts that upon film compression non-DPPC components are gradually excluded from the air-water interface into a surface-associated surfactant reservoir. However, detailed experimental evidence of the squeeze-out within the physiologically relevant high surface pressure range is still lacking. In the present work, we studied four animal-derived clinical surfactant preparations, including Survanta, Curosurf, Infasurf, and BLES. By comparing compression isotherms and lateral structures of these surfactant films obtained by atomic force microscopy within the physiologically relevant high surface pressure range, we have derived an updated squeeze-out model. Our model suggests that the squeeze-out originates from fluid phases of a phase-separated monolayer. The squeeze-out process follows a nucleation-growth model and only occurs within a narrow surface pressure range around the equilibrium spreading pressure of lung surfactant. After the squeeze-out, three-dimensional nuclei stop growing, thereby resulting in a DPPC-enriched interfacial monolayer to reduce the air-water surface tension to very low values.


Assuntos
Fosfolipídeos/química , Surfactantes Pulmonares/química , Membranas Artificiais , Tamanho da Partícula , Tensão Superficial
3.
Lab Chip ; 13(12): 2285-91, 2013 Jun 21.
Artigo em Inglês | MEDLINE | ID: mdl-23666050

RESUMO

An opto-thermocapillary micromanipulator (OTMm) capable of single-cell manipulation and patterning is presented here. The OTMm uses a near-infrared laser focused on an ITO substrate to induce thermocapillary convection that can trap and transport living cells with forces of up to 40 pN. The OTMm complements other cell-manipulation technologies, such as optical tweezers and dielectrophoresis, as it is less dependent upon the optical and electrical properties of the working environment, and can function in many types of cell culture media. The OTMm was used to construct single-cell matrices in two popular hydrogels: PEGDA and agarose. High viability rates were observed in both hydrogels, and cells patterned in agarose spread and migrated during subsequent culturing.


Assuntos
Micromanipulação/métodos , Pinças Ópticas , Animais , Hidrogéis/química , Camundongos , Micromanipulação/instrumentação , Células NIH 3T3 , Polietilenoglicóis/química , Temperatura
4.
Artigo em Inglês | MEDLINE | ID: mdl-24110933

RESUMO

Microrobots made of light-absorbing hydrogel material were actuated by optically induced thermocapillary flow and move at up to 700 µm/s. The micro-assembly capabilities of the microrobots were demonstrated by assembling polystyrene beads and yeast cells into various patterns on standard glass microscope slides. Two microrobots operating independently in parallel were also used to assemble micro-hydrogel structures.


Assuntos
Absorção de Radiação , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Luz , Robótica/métodos , Lasers , Microesferas , Poliestirenos/química , Saccharomyces cerevisiae/citologia
5.
ACS Nano ; 7(12): 10525-33, 2013 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-24266809

RESUMO

Interaction with the pulmonary surfactant film, being the first line of host defense, represents the initial bio-nano interaction in the lungs. Such interaction determines the fate of the inhaled nanoparticles and their potential therapeutic or toxicological effect. Despite considerable progress in optimizing physicochemical properties of nanoparticles for improved delivery and targeting, the mechanisms by which inhaled nanoparticles interact with the pulmonary surfactant film are still largely unknown. Here, using combined in vitro and in silico methods, we show how hydrophobicity and surface charge of nanoparticles differentially regulate the translocation and interaction with the pulmonary surfactant film. While hydrophilic nanoparticles generally translocate quickly across the pulmonary surfactant film, a significant portion of hydrophobic nanoparticles are trapped by the surfactant film and encapsulated in lipid protrusions upon film compression. Our results support a novel model of pulmonary surfactant lipoprotein corona associated with inhaled nanoparticles of different physicochemical properties. Our data suggest that the study of pulmonary nanotoxicology and nanoparticle-based pulmonary drug delivery should consider this lipoprotein corona.


Assuntos
Lipoproteínas/química , Nanopartículas/química , Surfactantes Pulmonares/química , Administração por Inalação , Adsorção , Animais , Produtos Biológicos/química , Bovinos , Simulação por Computador , Sistemas de Liberação de Medicamentos , Durapatita/química , Interações Hidrofóbicas e Hidrofílicas , Bicamadas Lipídicas/química , Lipídeos/química , Pulmão/efeitos dos fármacos , Simulação de Dinâmica Molecular , Nanotecnologia , Poliestirenos/química , Transporte Proteico
6.
Lab Chip ; 12(19): 3821-6, 2012 Oct 07.
Artigo em Inglês | MEDLINE | ID: mdl-22899225

RESUMO

A novel hydrogel microrobot made of poly(ethylene glycol) diacrylate (PEGDA) is reported. This disk-shaped microrobot is optothermally actuated by laser-induced bubbles. A pulsed laser is used to smoothly actuate an 80-µm-diameter bubble microrobot at up to 320 µm s(-1). A single microrobot or a pair of microrobots working in cooperation were used to assemble 20-µm-diameter polystyrene beads into different patterns. The microrobots were also used to assemble patterns made of single yeast cells and cell-laden agarose microgels. The patterned yeast cells and cell-laden microgels were cultured, and the cells successfully multiplied.


Assuntos
Gases/química , Hidrogel de Polietilenoglicol-Dimetacrilato/química , Robótica/instrumentação , Géis/química , Lasers , Polietilenoglicóis/química , Poliestirenos/química , Saccharomyces cerevisiae/citologia
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