Biomedical subjects

Timothy E McKnight

Publications and source records attributed to Timothy E McKnight.

4 recordsLinked to original sources

PubMed2007

Quantitative analysis of EDC-condensed DNA on vertically aligned carbon nanofiber gene delivery arrays.

Vertically aligned carbon nanofibers (VACNFs) with immobilized DNA have been developed as a novel tool for direct physical introduction and expression of exogenous genes in mammalian cells. Immobilization of DNA base amines to the carboxylic acids on nanofibers can influence the accessibility and transcriptional activity of the DNA template, making it necessary to determine the number of accessible gene copies on nanofiber arrays. Polymerase chain reaction (PCR) and in vitro transcription (IVT) were used to investigate the transcriptional accessibility of DNA tethered to VACNFs by correlating the yields of both IVT and PCR to that of non-tethered, free DNA. Yields of the promoter region and promoter/gene region of bound DNA plasmid were high. Amplification using primers designed to cover 80% of the plasmid failed to yield any product. These results are consistent with tethered, longer DNA sequences having a higher probability of interfering with the activity of DNA and RNA polymerases. Quantitative PCR (qPCR) was used to quantify the number of accessible gene copies tethered to nanofiber arrays. Copy numbers of promoters and reporter genes were quantified and compared to non-tethered DNA controls. In subsequent reactions of the same nanofiber arrays, DNA yields decreased dramatically in the non-tethered control, while the majority of tethered DNA was retained on the arrays. This decrease could be explained by the presence of DNA which is non-tethered to all samples and released during the assay. This investigation shows the applicability of these methods for monitoring DNA immobilization techniques.

Carbon↗

PubMed2006

Resident neuroelectrochemical interfacing using carbon nanofiber arrays.

Carbon nanofiber electrode architectures are used to provide for long-term, neuroelectroanalytical measurements of the dynamic processes of intercellular communication between excitable cells. Individually addressed, vertically aligned carbon nanofibers are incorporated into multielement electrode arrays upon which excitable cell matrixes of both neuronal-like derived cell lines (rat pheochromocytoma, PC-12) and primary cells (dissociated cells from embryonic rat hippocampus) are cultured over extended periods (days to weeks). Electrode arrays are characterized with respect to their response to easily oxidized neurotransmitters, including dopamine, norepinephrine, and 5-hydroxytyramide. Electroanalysis at discrete electrodes following long-term cell culture demonstrates that this platform remains responsive for the detection of easily oxidized species generated by the cultured cells. Preliminary data also suggests that quantal release of easily oxidized transmitters can be observed at nanofiber electrodes following direct culture and differentiation on the arrays for periods of at least 16 days.

Animals↗

PubMed2005

Synthetic nanoscale elements for delivery of materials into viable cells.

Arrays of vertically aligned carbon nanofibers (VACNFs) provide structures that are well suited for the direct integration and manipulation of molecular-scale phenomena within intact, live cells. VACNFs are fabricated via a combination of microfabrication techniques and catalytic plasma-enhanced chemical vapor deposition. In this chapter, we discuss the synthesis of VACNFs and detail the methods for introducing these arrays into the intracellular domain of mammalian cells for the purpose of delivering large macromolecules, specifically plasmid DNA, on a massively parallel basis.

Animals↗

PubMed2004

A magnetocaloric pump for microfluidic applications.

A magnetocaloric pump provides a simple means of pumping fluid using only external thermal and magnetic fields. The principle, which can be traced back to the early work of Rosensweig, is straightforward. Magnetic materials tend to lose their magnetization as the temperature approaches the material's Curie point. Exposing a column of magnetic fluid to a uniform magnetic field coincident with a temperature gradient produces a pressure gradient in the magnetic fluid. As the fluid heats up, it loses its attraction to the magnetic field and is displaced by cooler fluid. The impact of such a phenomenon is obvious: fluid propulsion with no moving mechanical parts. Until recently, limitations in the magnetic and thermal properties of conventional materials severely limited practical operating pressure gradients. However, recent advancements in the design of metal substituted magnetite enable fine control over both the magnetic and thermal properties of magnetic nanoparticles, a key element in colloidal-based magnetic fluids (ferrofluids). This paper begins with a basic description of the process and previous limitations due to material properties. This is followed by a review of existing methods of synthesizing magnetic nanoparticles as well as an introduction to a new approach based on thermophilic metal-reducing bacteria. We compare two compounds and show, experimentally, significant variation in specific magnetic and thermal properties. We develop the constitutive thermal, magnetic, and fluid dynamic equations associated with a magnetocaloric pump and validate our finite-element model with a series of experiments. Preliminary results show a good match between the model and experiment as well as approximately an order of magnitude increase in the fluid flow rate over conventional magnetite-based ferrofluids operating below 80 degrees C. Finally, as a practical demonstration, we describe a novel application of this technology: pumping fluids at the "lab-on-a-chip" microfluidic scale.

Computer-Aided Design↗