Biomedical subjects

Ian M Kennedy

Publications and source records attributed to Ian M Kennedy.

17 recordsLinked to original sources

PubMed2006

Optical characterization of eu-doped and undoped gd(2)o(3) nanoparticles synthesized by the hydrogen flame pyrolysis method.

Rare-earth-doped nanoparticles are promising materials for fluorescent labeling, as they are characterized by a high Stokes shift, narrow emission spectra, long lifetimes, minimized photobleaching, and low toxicity. We examined the structural and optical properties of europium-doped gadolinium oxide nanoparticles synthesized by the flame pyrolysis method, with specific emphasis on full spectral characterization and fluorescence kinetics. The emission-excitation characterization revealed the presence of predominantly monoclinic but also highly luminescent cubic phases with a prominent oxygen-to-europium charge-transfer band in the 230-260 nm range. A broad emission band in the visible region, corresponding to a similar band in undoped Gd(2)O(3), related to the matrix surface defects, was observed in time-gated spectroscopy of doped nanopowders. All of the examined nanopowders showed very short decay components, on the order of 2 ns, and much longer millisecond decay times characteristic of lanthanide ions. At intermediate times, on the order of 20-100 ns, a complex behavior of the decay was observed, indicative of progressive energy transfer to the lanthanide ion, which varied with different intrashell transitions. Structural characterization data by means of XRD measurements allowed for unambiguous determination of the Eu:Gd(2)O(3) crystallographic structure and cell dimensions to be consistent with a predominantly monoclinic phase.

Journal Article↗

PubMed2006

Eu3+-doped Gd2O3 nanoparticles as reporters for optical detection and visualization of antibodies patterned by microcontact printing.

Lanthanide oxide nanoparticles are promising luminescent probes in bioanalysis, because of their unique spectral properties, photostability, and low-cost synthesis. We report for the first time the application of europium-doped gadolinium oxide (Eu:Gd2O3) nanoparticles to the optical imaging of antibody micropatterns. The nanoparticles were synthesized by spray pyrolysis and coated with antibody (IgG) molecules by physical adsorption. Our experiments showed that the Eu:Gd2O3 is a good biocompatible solid support for antibody immobilization. The antibodies (anti-rabbit IgG) immobilized on the nanoparticles had excellent biological activity in the specific recognition reaction with rabbit IgG patterned in line strips (10 micromx10 microm) on a glass substrate by use of a micro-contact printing technique. The specific immunoreaction was confirmed by two independent microscopic techniques-fluorescence and scanning electron microscopy (SEM). Both microscopic images revealed that the nanoparticles were organized into designated structures as defined by the microcontact printing process with negligible non-specific binding. The nanoparticles can be used as fluorescent markers in a variety of immunosensing applications in a microscale format.

Adsorption↗

PubMed2005

Microarray immunoassay for phenoxybenzoic acid using polymer encapsulated Eu:Gd2O3 nanoparticles as fluorescent labels.

Currently, detection in microarray bioanalysis is based mainly on the use of organic dyes. To overcome photobleaching and spectral overlaps we applied a new type of fluorophore, crystalline europium-doped gadolinium oxide (Eu:Gd2O3) nanoparticles, as labels in immunoassay microarrays. The Eu:Gd2O3 nanoparticles synthesized by spray pyrolysis offer narrow red emission, large Stokes shift, photostable laser-induced fluorescence with a long lifetime (1 ms). The amino functionalization of the particles was achieved by poly(L-lysine) (PL) encapsulation. The formation of a stable PL shell was confirmed by TEM analysis, colloidal stability studies, and quantification of the surface reactive amino groups. The PL-encapsulated particles were covalently conjugated to antibodies and successfully applied as reporters in a competitive fluorescence microimmunoassay for phenoxybenzoic acid (PBA), a generic biomarker of human exposure to pyrethroid insecticides. Microarrays were fabricated by microcontact printing of BSA-PBA in line patterns (10 x 10 microm). Confocal fluorescence microscopy combined with internal standard (fluorescein) calibration was used for quantitative measurements. The microarray immunoassay demonstrated a limit of detection of 1.4 microg L(-1) PBA. This work suggests the potential application of lanthanide oxide nanoparticles as fluorescent probes in microarray and biosensor technology, immunodiagnostics, and high-throughput screening.

Antibodies↗

PubMed2004

Reduced lung cell proliferation following short-term exposure to ultrafine soot and iron particles in neonatal rats: key to impaired lung growth?

Particulate matter (PM) has been associated with a variety of negative health outcomes in children involving the respiratory system and early development. However, the precise mechanisms to explain how exposure to airborne particles may cause adverse effects in children are unknown. To study their influence on early postnatal development, a simple, laminar diffusion flame was used to generate an aerosol of soot and iron particles in the size range of 10 to 50 nm. Exposure of 10-day-old rat pups to soot and iron particles was for 6 h/day for 3 days. The lungs were examined following a single injection of bromodeoxyuridine (BrdU) 2 h prior to necropsy. Neonatal rats exposed to these particles demonstrated no effect on the rate of cell proliferation within terminal bronchioles or the general lung parenchyma. In contrast, within those regions arising immediately beyond the terminal bronchioles (defined as the proximal alveolar region), the rate of cell proliferation was significantly reduced compared with filtered air controls. These findings strongly suggest exposure to airborne particles during early neonatal life has significant direct effects on lung growth by altering cell division within critical sites of the respiratory tract during periods of rapid postnatal development. Such effects may result in altered growth in the respiratory system that may be associated with lifelong consequences.

Air Pollutants↗

PubMed2003

Fluorescence quenching competitive immunoassay in micro droplets.

A fluorescence quenching competitive immunoassay in micro droplets was applied to the sensitive detection of the pyrethroid insecticide, esfenvalerate. Laser induced fluorescence from rhodamine dye was used as a marker. The competitive immunoreaction was performed in micro droplets generated by a vibrating orifice aerosol generator system with a 10-microm diameter orifice. Fluorescence that was emitted from the droplets was detected by a 1/8 m imaging spectrograph with a 512 x 512 thermoelectrically cooled, charged-coupled device camera. The conjugate of esfenvalerate with rhodamine exhibited similar fluorescence to that of pure rhodamine 6G. When anti-esfenvalerate antibodies were added to the droplets, the fluorescence decreased. The reduction in emission was due to a strong quenching effect that arises from the interaction between the protein and rhodamine molecules following the antigen-antibody reaction. When a sample of esfenvalerate was added to the droplets, the release of the conjugated rhodamine from the antigen-antibody complex allowed the fluorescence signal to recover. An assay in a picoliter droplet sample was shown to enable detection down to approximately 0.1 nM. A very small mass of analyte could be detected with this method. A sample of river water was used to gauge the impact of matrix effects and was shown to give rise to negligible interference with the immunoassay.

Aerosols↗

PubMed2003

Oxidative stress and NFkappaB activation in the lungs of rats: a synergistic interaction between soot and iron particles.

Particulate matter (PM) has been associated with a variety of adverse health effects primarily involving the cardiopulmonary system. However, the precise biological mechanisms to explain how exposure to PM exacerbates or directly causes adverse effects are unknown. Particles of varying composition may play a critical role in these effects. To study such a phenomenon, a simple, laminar diffusion flame was used to generate aerosols of soot and iron particles in the ultrafine size range. Exposures of healthy adult rats were for 6 h/day for 3 days. Conditions used included exposure to soot only, iron only, or a combination of soot and iron. We found animals exposed to soot particles at 250 microg/m3 had no adverse respiratory effects. Exposure to iron alone at a concentration of 57 microg/m3 also had no respiratory effects. However, the addition of 45 microg/m3 of iron to soot with a combined total mass concentration of 250 microg/m3 demonstrated significant pulmonary ferritin induction, oxidative stress, elevation of IL-1beta, and cytochrome P450s, as well as activation of NFkappaB. These findings suggest that a synergistic interaction between soot and iron particles account for biological responses not found with exposure to iron alone or to soot alone.

Administration, Inhalation↗

PubMed2003

Competitive quenching fluorescence immunoassay for chlorophenols based on laser-induced fluorescence detection in microdroplets.

An improved biomonitoring system for the analysis of 2,4,6-trichlorophenol (TCP) in urine samples has been developed. The principle of the biosensor device is the detection of laser-induced fluorescence (LIF) in single microdroplets by a homogeneous quenching fluorescence immunoassay (QFIA). The competitive immunoassay occurs in microdroplets (d = 58,4 microm) produced by a piezoelectric generator system with 10-microm-diameter orifice. A continuous Ar ion laser (488 nm) excites the fluorescent tracer; its fluorescence is detected by a spectrometer attached to a 512 x 512 cooled, charge-coupled device camera. Fluorescence is quenched by specific binding of TCP polyclonal antibodies to the fluorescent tracer (hapten A-fluorescein); the quenching effect is diminished by the presence of the analyte. Thus, an increase in the signal is produced in a positive dose-dependent manner when TCP is present in the sample. In 10 mM PBS buffer, the IC50 of the LIF-microdroplet QFIA is 0.45 microg L(-1) reaching a LOD of 0.04 microg L(-1). The QFIA with the same reagents performed in microtiter plate format achieved a LOD of 0.36 microg L(-1) in buffer solution. Performance in human urine was similar to that observed in the buffer. A LOD of 1.6 ,g L(-1), with a dynamic range between 4 and 149.5 microg L(-1) in urine, was obtained without any sample treatment other than dilution with the assay buffer. The detectability achieved is sufficient for occupational exposure risk assessment.

Binding, Competitive↗

PubMed2003

Pulmonary responses of acute exposure to ultrafine iron particles in healthy adult rats.

As critical constituents of ambient particulate matter, transition metals such as iron may play an important role in health outcomes associated with air pollution. The purpose of this study was to determine the respiratory effects of inhaled ultrafine iron particles in rats. Sprague Dawley rats 10-12 weeks of age were exposed by inhalation to iron particles (57 and 90 microg/m(3), respectively) or filtered air (FA) for 6 h/day for 3 days. The median diameter of particles generated was 72 nm. Exposure to iron particles at a concentration of 90 microg/m(3) resulted in a significant decrease in total antioxidant power along with a significant induction in ferritin expression, GST activity, and IL-1beta levels in lungs compared with lungs of the FA control or of animals exposed to iron particles at 57 microg/m(3). NFkappaB-DNA binding activity was elevated 1.3-fold compared with that of control animals following exposure to 90 microg/m(3) of iron, but this change was not statistically significant. We concluded that inhalation of iron particles leads to oxidative stress associated with a proinflammatory response in a dose-dependent manner. The activation of NFkappaB may be involved in iron-induced respiratory responses, but further studies are merited.

Air Pollutants↗

PubMed2003

Microwave plasma conversion of volatile organic compounds.

A microwave-induced, steam/Ar/O2, plasma "torch" was operated at atmospheric pressure to determine the feasibility of destroying volatile organic compounds (VOCs) of concern. The plasma process can be coupled with adsorbent technology by providing steam as the fluid carrier for desorbing the VOCs from an adsorbent. Hence, N2 can be excluded by using a relatively inexpensive carrier gas, and thermal formation of oxides of nitrogen (NOx) is avoided in the plasma. The objectives of the study were to evaluate the technical feasibility of destroying VOCs from gas streams by using a commercially available microwave plasma torch and to examine whether significant byproducts were produced. Trichloroethene (TCE) and toluene (TOL) were added as representative VOCs of interest to a flow that contained Ar as a carrier gas in addition to O2 and steam. The O2 was necessary to ensure that undesirable byproducts were not formed in the process. Microwave power applied at 500-600 W was found to be sufficient to achieve the destruction of the test compounds, down to the detection limits of the gas chromatograph that was used in the analysis. Samples of the postmicrowave gases were collected on sorbent tubes for the analysis of dioxins and other byproducts. No hazardous byproducts were detected when sufficient O2 was added to the flow. The destruction efficiency at a fixed microwave power improved with the addition of steam to the flow that passed through the torch.

Adsorption↗

PubMed2002

Resonant Raman-scattering measurements of trichloroethene in a thermal boundary layer.

Concentration profiles of trichloroethene were measured in a boundary-layer flow over a heated ceramic surface. Raman scattering was excited with the fifth harmonic of a Nd:YAG laser at 213 nm. This wavelength took advantage of a resonance in the trichloroethene molecule to significantly enhance the C2HCl3 scattering cross section. The resonant Raman system was calibrated in a heated flow. The optical system was optimized so that measurements could be obtained close to the solid surface, normally a significant challenge for a spontaneous Raman-scattering setup. Measured concentrations indicated the lack of catalytic activity on a bare alumina surface. However, the results showed that a surface that was coated with Cr2O3-based zeolite was catalytically active.

Ceramics↗

PubMed1981