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Biomedical subjects

K M O'Brien

Publications and source records attributed to K M O'Brien.

At least 19 recordsLinked to original sources
PubMed2000

Quantification of diffusion distance within the spongy myocardium of hearts from antarctic fishes.

We developed a stereological method for quantifying diffusion distance within spongy myocardium. Using this method we compared the hearts of three species of Antarctic fishes that vary in expression of oxygen-binding proteins. We examined hearts from Gobionotothen gibberifrons, a red-blooded species whose ventricle has myoglobin (Mb), and hearts of two species of icefish that lack hemoglobin (Hb) and vary in expression of cardiac Mb; Chionodraco rastrospinosus expresses Mb, Chaenocephalus aceratus does not. Average diffusion distance within ventricular tissue is greater in red-blooded Antarctic teleosts (9.82 + or - 1.37 microm) compared with icefish (C. rastrospinosus, 6.20 microm + or - 0.86; C. aceratus, 6.23 + or - 0.41 microm). Average diffusion distance to a mitochondrion parallels this trend because mitochondria are uniformly distributed within cardiac muscle. Results show that loss of Hb is correlated with increased trabeculation of heart ventricle. Loss of Mb however, is not correlated with an increase in trabeculation of ventricular tissue, despite significant differences in cellular ultrastructure compared with species that express the protein.

Animals↗
PubMed2000

The interplay among cardiac ultrastructure, metabolism and the expression of oxygen-binding proteins in Antarctic fishes.

We examined heart ventricle from three species of Antarctic fishes that vary in their expression of oxygen-binding proteins to investigate how some of these fishes maintain cardiac function despite the loss of hemoglobin (Hb) and/or myoglobin (Mb). We quantified ultrastructural features and enzymatic indices of metabolic capacity in cardiac muscle from Gobionotothen gibberifrons, which expresses both Hb and Mb, Chionodraco rastrospinosus, which lacks Hb but expresses Mb, and Chaenocephalus aceratus, which lacks both Hb and Mb. The most striking difference in cellular architecture of the heart among these species is the percentage of cell volume occupied by mitochondria, V(v)(mit,f), which is greatest in Chaenocephalus aceratus (36.53+/-2.07), intermediate in Chionodraco rastrospinosus (20.10+/-0.74) and lowest in G. gibberifrons (15.87+/-0.74). There are also differences in mitochondrial morphologies among the three species. The surface area of inner mitochondrial membrane per volume of mitochondria, S(v)(imm, mit), varies inversely with mitochondrial volume density so that S(v)(imm,mit) is greatest in G. gibberifrons (29.63+/-1.62 microm(-)(1)), lower in Chionodraco rastrospinosus (21.52+/-0.69 microm(-)(1)) and smallest in Chaenocephalus aceratus (20.04+/-0.79 microm(-)(1)). The surface area of mitochondrial cristae per gram of tissue, however, is greater in Chaenocephalus aceratus than in G. gibberifrons and Chionodraco rastrospinosus, whose surface areas are similar. Despite significant ultrastructural differences, oxidative capacities, estimated from measurements of maximal activities per gram of tissue of enzymes from aerobic metabolic pathways, are similar among the three species. The combination of ultrastructural and enzymatic data indicates that there are differences in the density of electron transport chain proteins within the inner mitochondrial membrane; proteins are less densely packed within the cristae of hearts from Chaenocephalus aceratus than in the other two species. High mitochondrial densities within hearts from species that lack oxygen-binding proteins may help maintain oxygen flux by decreasing the diffusion distance between the ventricular lumen and mitochondrial membrane. Also, high mitochondrial densities result in a high intracellular lipid content, which may enhance oxygen diffusion because of the higher solubility of oxygen in lipid compared with cytoplasm. These results indicate that features of cardiac myocyte architecture in species lacking oxygen-binding proteins may maintain oxygen flux, ensuring that aerobic metabolic capacity is not diminished and that cardiac function is maintained.

Animals↗
PubMed1998

Computerized polymorphic marker identification: experimental validation and a predicted human polymorphism catalog.

A computational system for the prediction of polymorphic loci directly and efficiently from human genomic sequence was developed and verified. A suite of programs, collectively called POMPOUS (polymorphic marker prediction of ubiquitous simple sequences) detects tandem repeats ranging from dinucleotides up to 250 mers, scores them according to predicted level of polymorphism, and designs appropriate flanking primers for PCR amplification. This approach was validated on an approximately 750-kilobase region of human chromosome 3p21.3, involved in lung and breast carcinoma homozygous deletions. Target DNA from 36 paired B lymphoblastoid and lung cancer lines was amplified and allelotyped for 33 loci predicted by POMPOUS to be variable in repeat size. We found that among those 36 predominately Caucasian individuals 22 of the 33 (67%) predicted loci were polymorphic with an average heterozygosity of 0.42. Allele loss in this region was found in 27/36 (75%) of the tumor lines using these markers. POMPOUS provides the genetic researcher with an additional tool for the rapid and efficient identification of polymorphic markers, and through a World Wide Web site, investigators can use POMPOUS to identify polymorphic markers for their research. A catalog of 13,261 potential polymorphic markers and associated primer sets has been created from the analysis of 141,779,504 base pairs of human genomic sequence in GenBank. This data is available on our Web site (pompous.swmed.edu) and will be updated periodically as GenBank is expanded and algorithm accuracy is improved.

Computer Communication Networks↗
PubMed1997

Rescuing corrupted gel files from Model 377 and 373 DNA Sequencers.

Automated DNA sequencing requires the intensive use of computers to handle the large amount of data taken. When a computer failure occurs and the data are no longer accessible, all the expense and effort that went into the sequencing experiment is lost. By using the data storage architecture of Macintosh computers to our advantage, we may prevent this loss in the case of automatic sequencers from PE Applied Biosystems. The software required to allow the experimenter to do this has been written and is available free of charge.

Automation↗
PubMed1993

Cancer in the augmented breast. Diagnosis and prognosis.

BACKGROUND: The stage of breast cancer at diagnosis in women who have undergone augmentation mammoplasty has been examined with conflicting results. Additional evaluation of this population is needed. METHODS: A retrospective review of a large cancer center's breast cancer registry for a 116-month period was performed. Comparisons of patient age, method of detection, and pathologic stage of disease at presentation were performed for 33 patients with augmented breasts and 1735 patients with nonaugmented breasts who were treated. RESULTS: Twenty-four percent of the patients with augmented breasts and 42% of those with nonaugmented breasts had mammographically detected cancers (P = NS). The incidence of ductal carcinoma in situ in the two groups was similar (18% versus 15%). Sizes of the mammographically detected tumors in the two groups were comparable; however, palpable tumors in the augmented group were significantly smaller than were those in the nonaugmented group. Overall, a significant difference in axillary lymph node involvement was detected, with 19% of the augmented group and 41% of the nonaugmented group having positive lymph nodes. In patients with palpable tumors, axillary lymph node involvement also was significantly different, with 22% of the patients with augmented breasts and 58% of those with nonaugmented breasts having axillary lymph node metastases. In the mammographically detected tumors, there was no significant difference in axillary lymph node metastases between the patients with augmented (13%) and those with nonaugmented (15%) breasts. CONCLUSIONS: Early detection of breast cancer is possible in the patient who has undergone augumentation mammoplasty. Mammography was successful in detecting occult lesions, and palpation detected smaller tumors. In this cohort, there was no evidence to suggest that the patients with augmented breasts had more advanced disease at presentation.

Adult↗
PubMed1993

Photoinhibition of smooth muscle cell migration: potential therapy for restenosis.

Evidence from animal, autopsy, and atherectomy studies demonstrates that migration and proliferation of smooth muscle cells of medial origin result in neointima formation and decreased luminal cross-sectional area. The purpose of this study was to evaluate whether low energy light irradiation can inhibit smooth muscle cell migration and therefore potentially reduce the degree of neointima formation and the incidence of restenosis. The migration kinetics of bovine aortic smooth muscle cell monolayers were examined using a fence assay. The effect on smooth muscle cell migration of irradiation with monochromatic light at wave-lengths ranging from 400 to 700 nm was compared to the migration of cells irradiated with broadband white light or maintained in the dark. Wavelength specific photoinhibition of smooth muscle cell migration was observed; 594-600 nm light reproducibly inhibited migration by 12-29% (P < 0.05). Migration rate was significantly reduced following daily radiant exposures of 1.0 J/cm2 as well as following a single radiant exposure of 0.09 or 0.9 J/cm2. The decrease in migration was not associated with any change in cell proliferation or [3H] thymidine incorporation. We conclude that 594-600 nm light inhibits smooth muscle cell migration in vitro and may potentially be used in vivo to decrease fibrointimal thickening following arterial injury. This application of photoinhibition may be useful in retarding restenosis following angioplasty.

Animals↗
PubMed1992

Detection of calcified atherosclerotic plaque by laser-induced plasma emission.

The use of fluorescence spectroscopy to discriminate atherosclerotic from normal tissue is limited by a lower sensitivity for calcified than noncalcified atherosclerotic plaque (65% vs. 93%, respectively). To evaluate plasma emission as a means to detect calcified plaque, 325 normal and atherosclerotic cadaveric aortic sites were irradiated through a 100-micron silica fiber in blood by a pulsed holmium laser (lambda = 2.1 microns, fluence = 4 J/mm2). A photodiode positioned near the proximal end of the fiber detected plasma emission during a laser pulse. Plasma emission was detected at 0% (0/110) of normal, 0% (0/107) of noncalcified atherosclerotic tissue, and 91% (98/108) of calcified atherosclerotic sites. Spectroscopic analysis confirmed the presence of calcium lines in the plasma emission from calcified atherosclerotic plaque. Although ablative fluences (greater than 3 J/mm2) were required for plasma generation, a single laser pulse ablated only to a depth of 67 +/- 16 microns in normal tissue. In an additional 10 calcified atherosclerotic sites, laser ablation was continued as long as plasma emission was detected. In all cases, plaque ablation was terminated before arterial perforation. Furthermore, the adjunctive use of plasma detection improved the accuracy of fluorescence spectroscopic classification of normal and atherosclerotic tissue. In conclusion, plasma detection has a high sensitivity (91%) and specificity (100%) for calcified atherosclerotic plaque and may be a useful adjunct for laser angioplasty guidance. Furthermore, plasma detection can be implemented both simply and inexpensively.

Aluminum Silicates↗
PubMed1991

Design and evaluation of a fiberoptic fluorescence guided laser recanalization system.

Current angioplasty techniques for recanalization of totally occluded arteries are limited by the inability to cross the occlusion and by the risk of perforation. A fiberoptic fluorescence guided laser recanalization system was developed and evaluated in vitro for recanalization of 17 human femoral or tibial totally occluded arterial segments (length 1.9-6.8 cm, diameter 2.5-6.0 mm). A 400 or 600 micron silica fiber was coupled to a helium-cadmium laser (lambda = 325 nm) for fluorescence excitation and to a holmium: YAG laser (lambda = 2.1 micron) for tissue ablation. Fluorescence was recorded during recanalization after every other holmium laser pulse. During recanalization, each arterial segment was bent 30-90 degrees with respect to the fiber to simulate arterial tortuosity. Ablation continued with fiber advancement as long as the fluorescence confirmed that the target tissue was atherosclerotic. Arterial spectra were classified as normal or atherosclerotic by an on-line computerized fluorescence classification algorithm (sensitivity 93%, specificity 95%). Normal fluorescence necessitated redirection of the fiber greater than 30 times per segment to continue recanalization. Fifteen of 17 totally occluded arteries had multiple recanalization channels created following total energy delivery of 40-1,016 Joules per segment with no angiographic or histologic evidence of laser perforation. Two heavily calcified arterial occlusions were not recanalized due to inhibition of holmium: YAG laser ablation by the recording of normal fluorescence spectra. Therefore, this fluorescence guided laser recanalization system appears safe and effective for recanalization of totally occluded arteries and merits in vivo evaluation. However, the lower sensitivity of fluorescence detection of heavily calcified plaques may limit the efficacy (but not safety) of fluorescence guided recanalization of heavily calcified occlusions.

Angioplasty, Laser↗
PubMed1991

Neural network and conventional classifiers for fluorescence-guided laser angioplasty.

In laser angioplasty, fluorescence spectra of targeted tissue may be used to classify the tissue as atherosclerotic or normal and guide selective laser ablation of atherosclerotic plaque. Here, the ability of the back-propagation and K-nearest neighbors techniques to classify arterial fluorescence spectra is investigated. Both methods are competitive with other classification schemes. The relative performance of variations on both techniques is used to make inferences about the geometry of the classification task.

Angioplasty, Laser↗
PubMed1990

Expression of epidermal growth factor receptor sequences as E. coli fusion proteins: applications in the study of tyrosine kinase function.

To investigate the functions of key domains of the epidermal growth factor receptor (EGFR), various EGFR-derived peptide sequences were expressed in Escherichia coli as glutathione S-transferase (GST) fusion proteins. The purified fusion proteins (GST-TK0-8) were tested as substrates for the tyrosine kinase activities of the EGFR and c-src. Both the GST-TK4 fusion protein, which contains the major C-terminal tyrosine autophosphorylation sites of the EGFR, and GST-TK7, which contains the connecting sequence between the EGFR kinase domain and the C-terminal autophosphorylation domain, were strongly phosphorylated by the EGFR and c-src. Hence the candidate tyrosine phosphorylation sites present in the connecting sequences of the EGFR, as well as the known autophosphorylation sites of the EGFR, can be phosphorylated by the two tyrosine kinases. The protein GST-TK7 was phosphorylated by c-src with a KM of 5-10 microM, which indicated a potential interaction between the connecting segment of the EGFR and the c-src kinase. The GST fusion proteins were also used to map the sites recognized by two anti-EGFR monoclonal antibodies and a polyclonal serum raised against an EGFR tyrosine kinase domain fragment. The recognition site of one monoclonal antibody was determined to be in a short sequence surrounding tyr1068, a primary site of autophosphorylation in the C-terminal domain of the receptor. The anti-peptide polyclonal serum recognized only sequences in the GST-TK7 fusion protein, and hence binds to the connecting sequence between the kinase core and the C-terminal domain. These antibodies will therefore be useful reagents for studying the function of two key structural elements of the EGFR tyrosine kinase. The GST-TK fusion proteins should have many other applications in the study of EGFR catalysis and mitogenic signalling.

Amino Acid Sequence↗
PubMed1990

Characterization of the site dependency of normal canine arterial fluorescence.

The difference in fluorescence between normal and atherosclerotic artery has been proposed as a feedback mechanism to guide selective laser ablation of atherosclerotic plaque. This fluorescence difference is due to the relative difference in collagen:elastin content of normal artery and atherosclerotic plaque. However, normal arteries have site-dependent variation in collagen: elastin content which may affect their fluorescence spectra. To evaluate the site dependency of normal arterial fluorescence, helium-cadmium (325 nm) laser-induced fluorescence spectra were analyzed in vitro from the ascending aorta, abdominal aorta, and carotid, femoral, renal, and coronary arteries (N = 57) of 12 normal mongrel dogs. Elastin and collagen contents were determined for a subset of these arteries (N = 15). The spectral width of normal arterial Fluorescence varied by site and correlated with the measured collagen:elastin content at each site (r = -0.84, P less than 0.005). Fluorescence spectra were decomposed into collagen and elastin spectral components by using a linear model with a least-squared error criterion fit. The derived collagen and elastin spectral coefficients correlated with the measured collagen and elastin tissue content (r = 0.75 and 0.83 respectively, P less than 0.005). Thus, the fluorescence spectra of normal arteries is site dependent and correlates with the collagen:elastin content. Therefore, spectral feedback algorithms for laser angioplasty guidance must be site specific.

Animals↗
PubMed1990

Laser-induced fluorescence spectroscopy of human colonic mucosa. Detection of adenomatous transformation.

To evaluate the potential of laser-induced fluorescence spectroscopy for the detection of premalignant lesions of the gastrointestinal tract, the hypothesis that adenomatous transformation of colonic mucosa results in an alteration of laser-induced fluorescence that enables its differentiation from normal or hyperplastic tissue was tested. A fiberoptic catheter coupled to a helium-cadmium laser (325 nm) and an optical multichannel analyzer were used to obtain fluorescence spectra (350-600 nm) from 35 normal colonic specimens and 35 resected adenomatous polyps. A score based on six wavelengths was derived by stepwise multivariate linear regression analysis of the spectra. The mean score (+/- SEM) was + 0.86 +/- 0.06 for normal mucosa and -0.86 +/- 0.06 for adenomatous polyps (P less than 0.001). Spectra from an additional 34 normal specimens, 16 adenomatous polyps, and 16 hyperplastic polyps were prospectively classified with accuracies of 100%, 100%, and 94%, respectively. The mean score for hyperplastic polyps was significantly different from adenomatous (P less than 0.001) but not from normal tissue. Thus, quantitative analysis of fluorescence spectra enables the detection of adenomatous transformation in colonic mucosa.

Adenomatous Polyposis Coli↗
PubMed1989

Change in laser-induced arterial fluorescence during ablation of atherosclerotic plague.

Analysis of the change in arterial fluorescence during plaque ablation may provide the basis for developing a fluorescence-guided ablation system capable of selective plaque ablation without risk of vessel perforation. Accordingly, fluorescence spectra were recorded from 91 normal and 91 atherosclerotic specimens of cadaveric human aorta. The ratio of the laser-induced fluorescence intensity at 382 nm to 430 nm (LIF ratio) was capable of classifying these specimens with an 89% accuracy with a threshold value of 1.8 (atherosclerotic greater than or equal to 1.8, normal less than 1.8). To characterize the change in fluorescence during plaque ablation, mechanical plaque ablation with a cold microtome was performed on 16 atherosclerotic aortic specimens. Fluorescence spectra were recorded serially after each 100 microns of plaque ablation; recordings revealed a change in fluorescence spectra from atherosclerotic to a normal pattern. With an LIF ratio of 1.8 to signal termination of plaque ablation, 15 of the atherosclerotic plaques had a residual plaque thickness less than 200 microns; one specimen had a residual plaque thickness of 300 microns. No specimen demonstrated ablation of the media. There was a statistically significant correlation between LIF ratio and plaque thickness (r = .73, P less than .001), but considerable variation in LIF ratio existed at each thickness. Therefore, laser-induced fluorescence spectroscopy is capable of discriminating atherosclerotic from normal aorta and of signaling completion of plaque ablation.

Angioplasty, Balloon↗
PubMed1989

Fluorescence spectroscopy guidance of laser ablation of atherosclerotic plague.

Laser-induced fluorescence (LIF) spectroscopy can only be used for laser angioplasty guidance if high-power laser ablation does not significantly alter the pattern of tissue fluorescence. Although the spectra of normal and atherosclerotic arteries differ, the change in fluorescence spectra following laser angioplasty has not been well studied. Therefore, the purpose of this study was to assess whether laser-induced fluorescence spectroscopy could guide selective laser ablation of atherosclerotic plaque and, if so, to develop a quantitative LIF score that could be used to control a "smart" laser angioplasty system. Baseline LIF spectroscopy of 50 normal and 50 atherosclerotic human aortic specimens was performed using an optical fiber coupled to a He-Cd laser and optical multichannel analyzer. LIF was then serially recorded during erbium:YAG laser ablation of 27 atherosclerotic specimens. Laser ablation was terminated when the arterial LIF spectrum visually appeared normal. Histologic analysis revealed a mean initial plaque thickness of 1,228 +/- 54 microns and mean residual plaque thickness of 198 +/- 27 microns. Ablation of the media occurred in only three specimens. A discriminant function was derived to discriminate atherosclerotic from normal tissue for computer guidance of laser angioplasty. The LIF score, derived from stepwise multivariate linear regression analysis of the LIF spectra, correctly classified 93% of aortic specimens. The spectra obtained from the atherosclerotic specimens subjected to fluorescence-guided laser revealed a change in score from "atherosclerotic" to "normal" following plaque ablation. Seven atherosclerotic specimens were subjected to laser angioplasty with on-line computer control using the LIF score. Mean initial plaque thickness was 1,014 +/- 86 microns, and mean residual plaque thickness was 78 +/- 29 microns. There was no evidence of ablation of the media. Therefore, LIF guidance of laser ablation resulted in minimal residual plaque without arterial perforation. These findings support the feasibility of an LIF-guided laser angioplasty system for selective atherosclerotic plaque ablation.

Arteriosclerosis↗