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Biomedical subjects

P C Goodwin

Publications and source records attributed to P C Goodwin.

12 recordsLinked to original sources

Image metrics in the statistical analysis of DNA microarray data.

DNA microarrays represent an important new method for determining the complete expression profile of a cell. In "spotted" microarrays, slides carrying spots of target DNA are hybridized to fluorescently labeled cDNA from experimental and control cells and the arrays are imaged at two or more wavelengths. In this paper, we perform statistical analysis on images of microarrays and show that quantitating the amount of fluorescent DNA bound to microarrays is subject to considerable uncertainty because of large and small-scale intensity fluctuations within spots, nonadditive background, and fabrication artifacts. Pixel-by-pixel analysis of individual spots can be used to estimate these sources of error and establish the precision and accuracy with which gene expression ratios are determined. Simple weighting schemes based on these estimates are effective in improving significantly the quality of microarray data as it accumulates in a multiexperiment database. We propose that error estimates from image-based metrics should be one component in an explicitly probabilistic scheme for the analysis of DNA microarray data.

DNA, Complementary↗

Reliability of leg muscle electromyography in vertical jumping.

In this study we aimed to determine the reliability of the surface electromyography (EMG) of leg muscles during vertical jumping between two test sessions, held 2 weeks apart. Fifteen females performed three maximal vertical jumps with countermovement. The displacement of the body centre of mass (BCM), duration of propulsion phase (time), range of motion (ROM) and angular velocity of the knee and surface EMG of four leg muscles (rectus femoris, vastus medialis. biceps femoris and gastrocnemius) were recorded during the jumps. All variables were analysed throughout the propulsion and mid-propulsion phases. Intraclass correlation coefficients (ICC) for the rectus femoris, vastus medialis, biceps femoris and gastrocnemius were calculated to be 0.88, 0.70, 0.24 and 0.01, respectively. BCM, ROM and time values all indicated ICC values greater than 0.90, and the mean knee angular velocity was slightly lower, at 0.75. ICCs between displacement of the BCM and integrated EMG (IEMG) of the muscles studied were less than 0.50. The angular velocity of the knee did not correlate well with muscle activity. Factors that may have affected reliability were variations in the position of electrode replacement, skin resistance, cross-talk between muscles and jump mechanics. The results of this study suggest that while kinematic variables are reproducible over successive vertical jumps, the degree of repeatability of an IEMG signal is dependent upon the muscle studied.

Adult↗

Fluorescence image analysis of the association between Porphyromonas gingivalis and gingival epithelial cells.

We have developed a fluorescence imaging technique using a DNA-binding dye to visualize, over time, the physical interactions between Porphyromonas gingivalis and human gingival epithelial cells in vitro. The results extend previous observations of P. gingivalis invasion of gingival epithelial cells based on indirect measurements. An intracellular location for P. gingivalis was established by optical sectioning of images in the z-plane. Kinetic analysis showed that P. gingivalis invasion of epithelial cells is a rapid and efficient process, reaching completion after 12 min. Imaging of infected monolayers revealed that over 90% of a population of gingival epithelial cells contained bacteria. Furthermore, only vital bacteria were capable of invasion, and intracellular bacteria congregated in the perinuclear region of the epithelial cells. P. gingivalis remained inside the epithelial cells over a 24 h period and induced rearrangement of the actin cytoskeleton along with alteration of the size and shape of the epithelial cells. These findings provide direct evidence that entry rates of P. gingivalis into gingival epithelial cells are high and rapid, and that internalized bacteria initially localize in a specific region of the epithelial cells.

Cell Nucleus↗

Mitochondrial participation in the intracellular Ca2+ network.

Calcium can activate mitochondrial metabolism, and the possibility that mitochondrial Ca2+ uptake and extrusion modulate free cytosolic [Ca2+] (Cac) now has renewed interest. We use whole-cell and perforated patch clamp methods together with rapid local perfusion to introduce probes and inhibitors to rat chromaffin cells, to evoke Ca2+ entry, and to monitor Ca2+-activated currents that report near-surface [Ca2+]. We show that rapid recovery from elevations of Cac requires both the mitochondrial Ca2+ uniporter and the mitochondrial energization that drives Ca2+ uptake through it. Applying imaging and single-cell photometric methods, we find that the probe rhod-2 selectively localizes to mitochondria and uses its responses to quantify mitochondrial free [Ca2+] (Cam). The indicated resting Cam of 100-200 nM is similar to the resting Cac reported by the probes indo-1 and Calcium Green, or its dextran conjugate in the cytoplasm. Simultaneous monitoring of Cam and Cac at high temporal resolution shows that, although Cam increases less than Cac, mitochondrial sequestration of Ca2+ is fast and has high capacity. We find that mitochondrial Ca2+ uptake limits the rise and underlies the rapid decay of Cac excursions produced by Ca2+ entry or by mobilization of reticular stores. We also find that subsequent export of Ca2+ from mitochondria, seen as declining Cam, prolongs complete Cac recovery and that suppressing export of Ca2+, by inhibition of the mitochondrial Na+/ Ca2+ exchanger, reversibly hastens final recovery of Cac. We conclude that mitochondria are active participants in cellular Ca2+ signaling, whose unique role is determined by their ability to rapidly accumulate and then release large quantities of Ca2+.

Animals↗

Potential role of angiotensin II in noise-induced increases in inner ear blood flow.

Guinea pigs were exposed to 120 dB white noise for 30 min and evidenced a four-fold elevation in plasma concentration of the potent vasoconstricting hormone angiotensin II (AII). Anesthetized animals received intra-arterial injections of AII at doses that approximated the endogenous levels measured following noise exposure. A marked decrease in skin blood flow was observed with a concomitant increase in cochlear blood flow as measured by laser Doppler flowmeters. Increased cochlear blood flow appeared to be secondary to the increases in systemic blood pressure induced by AII. These findings suggest that cochlear blood flow may increase during periods of intense noise exposure.

Angiotensin II↗

Measuring cochlear blood flow by laser Doppler spectroscopy.

Cochlear blood flow (CBF) was studied with a commercially available laser Doppler system in 20 guinea pigs. The cochlea was exposed to permit placement of the laser Doppler probe over the intact lateral wall of the basal turn. Ketamine and xylazine were used for anesthesia, and blood pressure was monitored from the femoral artery. In some cases, skin blood flow was monitored with a second laser Doppler system, and cardiac output was monitored with an ultrasonic Doppler system placed over the right brachiocephalic artery. We found that the laser Doppler signal is composed primarily of blood flow supplied by the internal auditory artery. Local pressure on the contents of the internal auditory canal after occipital craniotomy was found to reduce CBF to 15% of its original value in a reversible fashion. There was no change in CBF after bilateral occlusion of the common carotid arteries. There appears to be a mechanism governing CBF that stabilizes its value in the face of changes in blood pressure and cardiac output. This is similar to the vascular behavior of the central nervous system. Through the use of positive airway pressure and blood removal at different rates, cardiac output could be depressed to varying degrees. The magnitude of decrease in CBF was clearly related to the rate at which cardiac output and blood pressure dropped. This was confirmed when intravenous phenylephrine was given in sequential and increasing doses. CBF increased as blood viscosity decreased, as expected according to the vascular behavior of the central nervous system.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Inner ear blood flow measured with a laser Doppler system.

Cochlear blood flow was studied in guinea pigs, using a laser Doppler system during exposure to methoxamine hydrochloride and phentolamine hydrochloride, local cooling, or terminal asphyxia. Measurements were made through the intact lateral wall of the first turn of the cochlea. Observed changes in relationship to simultaneous measures of skin blood flow and BP indicated that this system provides a dynamic measure of inner ear blood flow. This technique may provide a strategy to evaluate the role of blood flow in normal and pathologic cochlear function and the utility of presumed cochlear vasoeffective agents proposed for treatment of certain inner ear disorders.

Animals↗

Cochlear deoxyglucose uptake: relationship to stimulus intensity.

The relationship between stimulus intensity and the uptake of 2-deoxyglucose (2-DG) in the cochlea of the gerbil was studied using an autoradiographic technique. In silence, incorporation of labeled 2-DG into stria vascularis and the spiral ligament was significantly higher than for other inner ear structures. With increasing intensity of noise exposure, 2-DG uptake in the spiral ganglion and VIIIth nerve increased dramatically when compared to the lateral wall structures. In contrast, relative 2-DG uptake in the organ of Corti was much less affected by noise exposure. Only at 105 dB SPL, the highest intensity tested, was a modest but statistically significant increase observed in the sensory epithelium. The small change in relative 2-DG uptake observed in the organ of Corti during acoustic stimulation is consistent with Davis' (1965: Quant. Biol. 30, 181-190) battery model of the cochlear transduction process. Alternatively, a larger change may have occurred, but been restricted to a small portion of the epithelium, such as one or both populations of hair cells.

Acoustic Stimulation↗

Laser Doppler measurements of cochlear blood flow.

Blood flow to the inner ear was studied with a laser Doppler system in the acute guinea pig. Flow was measured through the lateral wall of the basal turn. Changes in simultaneous measures of cochlear and skin flow with rebreathing, epinephrine, phentolamine and terminal bleeding were studied. Cochlear blood flow followed skin blood flow in most cases; where it did not, the change was in an expected direction. The laser Doppler flowmeter appears to provide a direct, dynamic and linear measure of inner ear blood flow.

Animals↗

The laser Doppler: a non-invasive measure of cochlear blood flow.

The present investigation demonstrates the utility of the laser Doppler flowmeter to provide a measure of cochlear blood flow dynamics. Cochlear and cutaneous blood flow were compared with arterial blood pressure during and following exposure to Angiotensin II, 5% carbon monoxide, 100% oxygen, mannitol, and saline. The observations indicate that: 1) cochlear blood flow generally parallels cutaneous blood flow; however, 2) when cutaneous beds vasoconstrict (e.g., AII, alpha-agonists), cochlear blood flow parallels blood pressure; and, 3) under the influence of agents that affect peripheral and central circulation (5% CO, 100% O2), cochlear blood flow may dissociate from cutaneous blood flow and blood pressure. The implications of these findings are discussed in terms of local control mechanisms that may be involved in the inner ear vasculature.

Angiotensin II↗