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Biomedical subjects

C H Barlow

Publications and source records attributed to C H Barlow.

At least 19 recordsLinked to original sources
PubMed1990

Time resolved 3-dimensional recording of redox ratio during spreading depression in gerbil brain.

Optical fluorescence and reflectance measurements have been used to map the distribution of metabolic states in three dimensions in the gerbil brain with a spatial resolution of 200 microns an a time resolution of 4-6 s. In Mongolian gerbils anesthetized with pentobarbital, the redox states of the nicotinamide adenine dinucleotide (NADH) and flavoprotein components of the electron transport chain exhibit two distinct phases during the wave of spreading depression: (1) a transient period of oxidation and (2) a prolonged period of reduction, during which the cytochromes are reduced, and the hemoglobin is predominantly in the deoxy form. These data are interpreted as indicating that the energy demand placed on the gerbil brain during such spreading depression wave is sufficient to drive the brain temporarily hypoxic.

Animals↗
PubMed1984

Delineation of myocardial ischemia in an isolated blood-perfused rabbit heart preparation.

Isolated perfused heart preparations provide controlled conditions for the study of myocardial respiration and metabolism. Most studies utilize a hemoglobin-free perfusate which requires high arterial oxygen tension and high coronary perfusion rate. A blood-perfused rabbit heart preparation using an "assist" rabbit to maintain blood homeostasis has been developed which is suitable for respiratory, metabolic, and spectroscopic studies of myocardial function under controlled conditions. Fluorescence emission of reduced nicotinamide adenine dinucleotide (NADH) from the surface of blood-perfused rabbit heart was photographed to delineate areas of myocardial ischemia detectable as NADH fluorescence from reduced mitochondria. Blood reperfusion of ischemic myocardium resulted in disappearance of NADH fluorescence. Reocclusion of the coronary artery resulted in the same pattern and amplitude of NADH fluorescence.

Animals↗
PubMed1982

Mechanism of action of hyaluronidase in decreasing myocardial ischemia post coronary occlusion in the isolated perfused rabbit heart.

The influence of hyaluronidase (H) on subacute experimental myocardial ischemia was studied in isolated perfused rabbit hearts. Changes in ischemic area were assessed by epicardial nicotinamide adenine dinucleotide (NADH) fluorescence photography, an intrinsic high-resolution display of myocardial ischemia. Computerized determination of ischemic area was made from standardized photographs. Hyaluronidase was begun 20 minutes after coronary artery occlusion at 4 units/ml perfusate. NADH fluorophotographs were taken at 10-minute intervals up to 60 minutes of ischemia. Coronary sinus oxygen tension (PcsO2), myocardial oxygen consumption (MVO2), and coronary flow were determined. After 70 minutes, the hearts were perfused with rhodamine solution to identify areas of myocardial perfusion. In 13 H-treated hearts 54.3% +/- 3.7% (mean +/- SEM) of the nonperfused area (rhodamine stained) was ischemic (NADH fluorescent). In 14 untreated hearts 79.8% +/- 3.2% of the nonperfused area was ischemic (p less than 0.0001) and the ischemic areas were uniform. The distance between perfused and ischemic tissue was 952 +/- 78 micrometers in the H hearts and 504 +/- 35 micrometers in the untreated heart (p less than 0.0001). In the H hearts PcsO2 increased to 155% of the post-ligation control while it decreased to 79% in the untreated hearts (p less than 0.0001). MVO2 decreased in the H-treated hearts to 62%; the untreated hearts had no further change. In the H-treated hearts, coronary flow increased to 146% of the post-ligation control while it fell to 91% in the untreated group (p less than 0.0001). We conclude that H increases coronary flow while decreasing MVO2 during subacute ischemia. In H-treated hearts, significant amounts of myocardium remain normoxic within the nonperfused areas, and may potentially be salvaged after prolonged myocardial ischemia.

Animals↗
PubMed1982

Optical measurements of oxygen delivery and consumption in gerbil cerebral cortex.

Oxygen metabolism of the cerebral cortex of anesthetized gerbils was monitored by surface fluorescence and reflectance spectrophotometry both in vivo and in specimens freeze-trapped by the surface-freezing technique. Fiber optic light guides were used for optical coupling for determining mitochondrial flavoprotein and pyridine nucleotide fluorescence and for recording dual-wavelength reflectance spectra that indicated the levels of ferrocytochromes aa3 and c + c1 in the cortex and the oxygen saturation of cortical hemoglobin. During anoxic episodes increases in ferrocytochromes aa3 and c + c1 and in reduced pyridine nucleotides and flavoproteins were observed only when the cortical hemoglobin was more than 85% disoxygenated. Indeed the steady-state levels of oxidation-reduction of mitochondrial respiratory-chain components remained constant over a wide range of oxygen delivery, increased reduction being observed only when the fraction of inspired oxygen fell below 6%; these properties of mitochondria in vivo resemble those found in vitro. However, in normoxic animals the absorbance at 605 nm that may arise from ferri- or ferrocytochrome aa3 is larger than would be expected of purified mitochondria and may represent a pool of mitochondria that remains reduced at all levels of tissue oxygenation or a pigment not involved in oxygen metabolism.

Animals↗
PubMed1981

Early ischemia after complete coronary ligation in the rabbit, dog, pig, and monkey.

The character and extent of the myocardial ischemic "borderzone" was assessed in the rabbit, dog, pig, and monkey. A fluorophotographic technique permitting high resolution (+/- 50 micrometers) display of myocardial ischemia has been developed. Reduced intracellular NADH (ischemia) fluoresces and may be photographed while oxidized NAD (perfused tissue) does not. A coronary artery was ligated for 5 min in open-chest rabbits, dogs, pigs, and monkeys. A fluorescent dye was injected into the left atrium as a coronary vascular marker, and the tissue was quick-frozen. The ischemic margin was well seen and was jagged in all species. The distance from anoxic to perfused tissue (borderzone) was less than 50 micrometers in all species. A narrow "oxygen-diffusion zone" of nonperfused non-anoxic tissue is visible in isolated heart perfused with blood-free solution. The width of this zone is inversely related to myocardial oxygen consumption and is less than 50 micrometers in a working blood-perfused heart. We have not yet correlated the oxygen diffusion zone with the clinically defined salvageable borderzone. In dogs, collateral vessels provide a heterogeneous border to the ischemic region so that the canine ischemic pattern differs from that of pigs, rabbits, and monkeys.

Animals↗
PubMed1980

Quantification of tissue fluorescein delivery and prediction of flap viability with the fiberoptic dermofluorometer.

Quantitative fluorescence assessment with the fiberoptic dermofluorometer has been introduced to overcome the inadequacies of ultraviolet inspection following intravenous administration of fluorescein. Initial applications in the pedicle flap of the rat have indicated that this method can predict flap viability with reproducible precision and accuracy. This minimally invasive technique should be of considerable assistance to the evaluation of hypoperfused states in laboratory and clinical situations.

Animals↗
PubMed1980

Surface fluorescence of reduced pyridine nucleotide of the perfused rat kidney: interrelation between metabolic and functional states.

Surface fluorescence of reduced pyridine nucleotide (PN) was recorded continously with a DC fluorormeter and correlated with changes in experimental conditions. As a light source for fluorescence excitation, an Hg arc lamp with a 340-375 nm filter in front was used; the fluorescence response of reduced PN was measured at 450-510 nm. The DC fluormeter and the Hg arc lamp were connected to the kidney by a trifurcated fiber optics light guide. Reduced PN fluorescence emission was corrected for changes in tissue opacity by a 1:1 subtraction of reflectance changes at 340-375 nm from the fluorescence (PN-deltaR). To obtain further information about the PN redox state of the total surface area of kidney cortex and to evaluate whether certain areas were insufficiently perfused, fluorescence photographs of the total surface area were taken. The results demonstrate that the described method is simple and provides specific information about the mitochondrial oxidation reduction state.

Animals↗
PubMed1980

Heterogeneity of the coronary microcirculation during low-flow ischemia: a model for the heart in shock.

The etiology of myocardial damage in shock is multifactorial. Poor perfusion of the coronary bed during shock may result in areas of cellular anoxia. This study examines the response of isolated perfused rabbit hearts to low-flow ischemia utilizing a high resolution assay of ischemia, NADH fluorescence photography. The hearts were made ischemic by reducing coronary perfusion pressure to 50% for 60 seconds. Islands of ischemic anoxia appear over the surface of the ventricle. The intensity of the ischemic response increased with successive ischemic insults (P less than 0.01). The number of anoxic islands can be reduced by pretreatment with papaverine (P less than 0.01). Anoxic islands are stable up to 20 minutes of ischemic time and are not stained by fluorescein angiography. The data suggest that an intrinsic inequality exists in the perfusion of the coronary microcirculation during low-flow ischemia.

Animals↗
PubMed1979

The nature of the copper atoms of cytochrome c oxidase as studied by optical and x-ray absorption edge spectroscopy.

X-ray absorption edge spectroscopy has been used to study the copper of 1--2 mM cytochrome c oxidase in the resting oxidized, mixed-valence, and fully reduced states. A comparison was made of this protein with copper complexes and with natural and artificial copper proteins. Spectra were obtained with synchrotron radiation from the SPEAR storage ring using highly sensitive fluorescence detectors. Temperatures of -80 to -120 degrees C were employed further to improve the stability of the samples and to avoid the possibility of either auto- or photon-induced reduction of the materials, which might have occurred in previous studies. In order to characterize the valence states of the Cu and Fe components, the samples were monitored by infrared and visible spectroscopy before and after irradiation by the X-ray beam. The combination of the optical and X-ray absorption techniques has afforded a deconvolution of the four species of copper in the various states of cytochrome c oxidase and the tentative assignment of Cu alpha, the copper redox coupled to the heme alpha of cytochrome alpha, as a highly covalent type of copper and Cu alpha 3, the copper of cytochrome alpha 3, as a more ionic 'blue' type I copper. The implications of these findings upon the mechanism of action of cytochrome oxidase are briefly outlined.

Absorptiometry, Photon↗
PubMed1979

Temporal relation between onset of cell anoxia and ischemic contractile failure. Myocardial ischemia and left ventricular failure in the isolated, perfused rabbit heart.

Contractile dysfunction is characteristic of the acutely ischemic myocardium. This study was undertaken to assess the temporal relations between the onset of cell anoxia and ischemic contractile failure in isolated, isovolumetric contracting rabbit hearts. High speed epicardial fluorescence photography using reduced nicotinamide adenosine nucleotide (NADH) was used to identify areas of cell anoxia. The onset of ischemia was correlated with deterioration of pressure generation over the course of sequential 60 second coronary arterial occlusions. In the isovolumetric contracting rabbit heart, areas of ischemia were detected 2 seconds after coronary occlusion. Significant reduction in peak systolic pressure occurred at 6 seconds of ischemic time and pressure continued to decrease throughout the 60 second period of coronary occlusion. NADH accumulation indicates imbalance of myocardial oxygen supply and demand and the cessation of oxygen utilization by the mitochondria. The results of this study indicate that ischemia is detectable within 1 to 2 seconds after coronary occlusion and that ischemic ventricular dysfunction occurs several seconds thereafter. Myocardial oxygen reserve is negligible.

Animals↗
PubMed1979

Epicardial ischemia as delineated with epicardial S-T segment mapping andnicotinamide adenine dinucleotide (NADH) fluorescence photography.

In isolated rabbit hearts with an experimental coronary arterial occlusion, epicardial ischemia was identified by reduced nicotinamide adenine dinucleotide (NADH) fluorescence photography, a technique that detects areas of myocardial anoxia. Epicardial S-T segment mapping was performed to evaluate the S-T segment changes across an ischemic border defined by NADH fluorescence. After S-T segment mapping and perfusion with a fluorescein dye, serial selections of the hearts revealed that the ischemic area was transmural and and the border was nearly perpendicular to the epicardial surface. As the epicardial ischemic border was approached, S-T segment elevation was first detected 3.3 mm outside the ischemic border, and increased over a transition zone 7 mm wide. S-T segment negativity was not detected immediately outside the ischemic border. It is concluded from these studies that S-T segment changes give relatively imprecise definition of an ischemic border, and that S-T segment changes across an ischemic border are not consistent with those predicted by solid angle analysis.

Animals↗
PubMed1979

Fluorescence mapping of mitochondrial redox changes in heart and brain.

Fluorescence techniques may be utilized to map changes in the distribution of mitochondrial redox states in heart and brain during ischemic or hypoxic stress. The basis of these techniques is the intrinsic fluorescence of reduced NADH and oxidized flavoprotein in mitochondria which respond to changes in critical oxygen supply. Ischemic areas in rabbit hearts induced by coronary ligation were detected and mapped based on the increase in NADH fluorescence in the ischemic zone. The width of the jeopardized normoxic tissue surrounding the ischemic area (less than 50--350 mu) was measured by combination of fluorescein angiography and NADH fluorescence. Areas of increased NADH fluorescence in gerbil brains after carotid artery ligation or induction of spreading depression were mapped in a similar manner. Intraoperative monitoring of flavoprotein fluorescence from human cerebral cortex after superficial temporal artery middle cerebral artery (STA-MCA) anastomoses demonstrated increased rates of cortical oxidative metabolism after the surgical procedures.

Animals↗