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Biomedical subjects

J G Miller

Publications and source records attributed to J G Miller.

At least 127 records · Page 7Linked to original sources

Anisotropy of the ultrasonic backscatter of myocardial tissue: II. Measurements in vivo.

The purpose of this investigation was to determine the angular dependence of the backscatter from canine myocardial tissue in vivo and to compare it with the variation of backscatter over the cardiac cycle that has been recognized and reported previously. The backscatter was measured from regions of left ventricular wall in canine hearts in which the fibers of the muscle lay parallel to the surface of the heart and were oriented predominantly in a circumferential fashion. Because of technical considerations, the angle of insonification was varied systematically through two cycles in which the angle relative to the muscle fiber axes ranged from 60 degrees-120 degrees. Backscatter was maximum at angles of interrogation perpendicular to the myocardial fibers and minimum at those most acute (60 degrees) relative to the orientation of the fibers. The previously observed variation of integrated backscatter over the heart cycle was evident at each angle of interrogation. At end systole, the average maximum-to-minimum angular variation of integrated backscatter as 5.0 +/- 0.4 dB. At end diastole, the average maximum-to-minimum angular variation was 3.2 +/- 0.4 dB. Thus, even though angular dependence of the backscatter from tissues with directionally oriented structures is substantial, the anisotropy does not account for cardiac-cycle-dependent variation of backscatter. Accordingly, the angular dependence should be incorporated in approaches to quantitative tissue characterization with ultrasound.

Animals↗

Effects of cage beddings on microsomal oxidative enzymes in rat liver.

The purpose of the present studies was to evaluate the effects of some commercially available cage beddings on rat liver microsomal cytochrome P-450-dependent drug-metabolizing enzyme, ethylmorphine N-demethylase, and the carcinogen-metabolizing enzyme, benzo(a)pyrene hydroxylase. Sprague-Dawley rats were housed in cages containing cedar chip, corncob or heat-treated pinewood bedding for 3 weeks. Control rats were housed in cages on wire bottom floors containing no bedding material. Rats housed in cages containing cedar chip showed 18, 46 and 49% increases in liver cytochrome P-450 content, ethylmorphine N-demethylase and benzo(a)pyrene hydroxylase activities, respectively. The liver enzyme activities of rats housed in cages containing corncob bedding were similar to those obtained with control rats. In contrast, the pinewood-bedded rats showed a 21% decrease in ethylmorphine N-demethylase activity without affecting cytochrome P-450 content and benzo(a)pyrene hydroxylase activity. Hexobarbital-induced sleep times of the variously bedded rats were similar to those of control animals. These data suggest that the commercial bedding materials differ in their abilities to affect liver microsomal enzymes. Thus, interlaboratory variability in basal enzyme activities reported in the literature may be partly due to bedding materials used in the animal's cages.

Animals↗

Relative roles of gap junction channels and cytoplasm in cell-to-cell diffusion of fluorescent tracers.

Intercellular (tissue) diffusion of molecules requires cytoplasmic diffusion and diffusion through gap junctional (or cell-to-cell) channels. The rates of tissue and cytoplasmic diffusion of fluorescent tracers, expressed as an effective diffusion coefficient, De, and a cytoplasmic diffusion coefficient, Dcyt, have been measured among the developing epidermal cells of a larval beetle, Tenebrio molitor L., to determine the contribution of the junctional channels to intercellular diffusion. Tracer diffusion was measured by injecting fluorescent tracers into cells and quantitating the rate of subsequent spread into adjacent cells. Cytoplasmic diffusion was determined by fluorescence photobleaching. These experiments show that gap junctional channels constitute approximately 70-80% of the total cell-to-cell resistance to the diffusion of organic tracers at high concentrations in this tissue. At low concentrations, however, the binding of tracer to cytoplasm slows down the cytoplasmic diffusion, which may limit intercellular diffusion.

Animals↗

Amino acid content of preimplantation rabbit embryos and fluids of the reproductive tract.

As a companion to amino acid transport and protein synthetic studies, it was of interest to quantify the amino acid pools in embryos and reproductive tract fluids during preimplantation development. Primary amines in the acid-soluble extracts of embryo and fluid samples were separated by high-performance liquid chromatography, reacted with o-phthalaldehyde, and quantified by fluorescence emission. The amino acid compositions of embryos were like those of corresponding reproductive tract fluids. Taurine was high in eggs and fluids but declined with development, while glycine levels rose. Glycine was highest in concentration in all samples (except the egg), followed by glutamate and alanine, while most other amino acids were consistently of low abundance.

Amino Acids↗

Quantitative ultrasonic tissue characterization with real-time integrated backscatter imaging in normal human subjects and in patients with dilated cardiomyopathy.

We have shown previously that the physical properties of myocardium in dogs can be characterized with quantitative ultrasonic integrated backscatter and that interrogation of the tissue with ultrasound can delineate cardiac cycle-dependent changes in ultrasonic backscatter in normal tissue that disappear with ischemia and reappear with reperfusion if functional integrity is restorable. To determine whether this approach can be applied to man, we implemented an automatic gain compensation and continuous data acquisition system to characterize myocardium with quantitative ultrasonic backscatter and to detect cardiac cycle-dependent changes in real time. We developed a two-dimensional echocardiographic system with quantitative integrated backscatter imaging capabilities for use in human subjects that can automatically differentiate ultrasonic signals from blood as opposed to those obtained from tissue and adjust the slope of the gain compensation appropriately. Real-time images were formed from a continuous signal proportional to the logarithm of the integrated backscatter along each A-line. In our initial investigation, 15 normal volunteers (ages 17 to 40 years, heart rates 44 to 88 beats/min) and five patients with dilated cardiomyopathy (ages 22 to 52, heart rates 82 to 120 beats/min) were studied with conventional parasternal long-axis echocardiographic views. Diastolic-to-systolic variation of integrated backscatter in the interventricular septum and left ventricular posterior wall was seen in each of the normal subjects averaging 4.6 +/- 1.4 dB (SD) and 5.3 +/- 1.5 dB (n = 127 sites), respectively.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Quantitative ultrasonic characterization of the nature of atherosclerotic plaques in human aorta.

To determine the feasibility of quantitative ultrasonic techniques to define the composition of atherosclerotic plaques, samples of freshly excised human aortas were sewn to a sample holder and immersed in a saline bath for ultrasonic interrogation. Integrated backscatter of a 2-microsecond portion of the backscattered radiofrequency signal was measured with a 10 MHz focused transducer. Integrated backscatter was calculated by normalizing the root-mean-square voltage of the gated signal to the root-mean-square voltage obtained by replacing the tissue sample with a nearly perfect reflector. Microscopic examination of the 124 interrogated sites allowed differentiation of normal, fibrous, fibrofatty, and calcified regions. A site was considered calcified if it contained any histochemically detectable calcium. Values of integrated backscatter were markedly elevated from calcified regions (-30.0 +/- 6.4 dB, n = 25; mean +/- SD; p less than 0.001) compared to normal (-43.2 +/- 2.4 dB, n = 20) and fibrofatty (-43.9 +/- 3.4 dB, n = 43) sites. Values from fibrous regions (-40.7 +/- 3.8 dB, n = 36) were also significantly different compared with the calcified and fibrofatty regions (p less than 0.001). Thus, we have demonstrated that quantitative ultrasonic backscatter identifies and differentiates calcification and fibrosis in atherosclerotic sites offering promise for the noninvasive assessment of the pathologic status of the arterial wall.

Aorta↗

Evidence for nystatin micelles in L-cell membranes from fluorescence photobleaching measurements of diffusion.

Diffusion of a nitrobenzoxadiazole derivative of the polyene antibiotic nystatin in the membranes of L cells is found to depend on the concentration of nystatin in the membrane. Its diffusion coefficient measured by fluorescence photobleaching decreases hyperbolically as the concentration of nystatin is increased. This behavior is reproduced when the concentration of the derivative is increased. In contrast, diffusion of a nitrobenzoxadiazole derivative of a phospholipid is insensitive to the nystatin concentration under these conditions. The nystatin-specific diffusion changes can be understood if nystatin exists in a monomer-micelle equilibrium within the membrane but cannot be accounted for by binding or phase partitioning.

Animals↗

Purines, prostaglandins and peptides--nature and cellular mechanisms of action of local assist and assassin agents in the ovary.

The evidence for a paracrine, progonadotropic role of adenosine in ovarian cells is summarized along with a capsule review of the origin and mechanisms of release and action of adenosine in other tissues. Briefly, adenosine markedly amplified rat and human luteal cell cyclic AMP and progesterone accumulation in the presence, but not the absence, of LH. The site of action of adenosine was found to be intracellular, linked to its phosphorylation, which resulted in increased levels of ATP. In rat luteal cells, adenosine blocked the acute antigonadotropic (luteolytic) action of PGF2 alpha. In the follicle, adenosine release from granulosal cells appeared to be stimulated by FSH. Adenosine and a nonmetabolized adenosine analog, augmented FSH-dependent inhibition of oocyte maturation in the presence or absence of an adenosine transport inhibitor. Inhibition of oocyte maturation by adenosine thus appears to be mediated by extracellular purinergic receptors. Paracrine, antigonadotropic agents also appear to regulate ovarian function. For example, GnRH elicits antigonadotropic activity in rat granulosal and luteal cells. We describe a novel, GnRH-like, ovarian hormone (GLOH) which may be the physiological ligand whose action GnRH mimics in rat ovarian cells. This protein was shown to be distinctly different from GnRH and a variety of other cyclic and noncyclic peptides. PGF2 alpha is a well known leutolytic agent and a summary of the antigonadotropic mechanism of PGF2 alpha action in rat luteal cells is presented. In these cells, the action of GnRH (or possibly the GnRH-like protein) and PGF2 alpha are mediated by separate membrane receptors but they appeared to share the same intracellular second messenger. Evidence for a role of products of phosphoinositol as a mediator of these antigonadotropic agents is summarized. We suggest that the ultimate mediator of antigonadotropic agents is Ca2+ which is released in the luteal cell in response to the intracellular mediator of antigonadotropic agents. For example, pharmacological agents which increase intracellular levels of Ca2+, mimicked the antigonadotropic action of GnRH and PGF2 alpha in rat luteal cells. Also, Ca2+ directly inhibited LH-sensitive adenylate cyclase activity in isolated luteal membranes, a paradigm in which GnRH and PGF2 alpha were inactive. The mechanism of Ca2+ action appeared to be linked to interference with GTP activation of adenylate cyclase. However, removal of extracellular Ca2+ did not abrogate the action of either GnRH or PGF2 alpha.(ABSTRACT TRUNCATED AT 400 WORDS)

Adenosine↗

Oocyte maturation is inhibited by adenosine in the presence of follicle-stimulating hormone.

Considerable evidence implicates cyclic 3', 5' adenosine monophosphate (AMP) in the maintenance of meiotic arrest of mammalian oocytes. Since this laboratory previously found that adenosine augmented follicle-stimulating hormone (FSH)-stimulated accumulation of cyclic AMP in oocyte-cumulus-complexes (OCC), in the present studies we investigated the possibility that adenosine inhibits maturation of oocytes. In rat OCC cultured in the presence of FSH, adenosine markedly inhibited oocyte maturation in a dose-dependent and biphasic manner. Maximum inhibition of oocyte maturation was seen with 1-30 microM adenosine in the presence of FSH, and half-maximal inhibition occurred with less than 0.3 microM adenosine. High levels of adenosine (100 microM) did not inhibit oocyte maturation in the presence of FSH. In the absence of FSH, adenosine showed little effect on oocyte maturation in the present studies, but increased the maximum inhibition of oocyte maturation produced by FSH approximately twofold. Like adenosine, adenosine triphosphate (ATP), adenosine diphosphate (ADP), and adenosine 5'-monophosphate (AMP) also inhibited oocyte maturation; whereas adenine, guanosine, inosine, and hypoxanthine were inactive at equivalent levels. The metabolism-resistant adenosine analog (2-chloroadenosine) was as active an inhibitor as adenosine. Inhibition produced by the adenine nucleotides may have been direct or due to conversion to adenosine by extracellular nucleotidases. The concentration dependence and purine specificity for inhibition of oocyte maturation are characteristic of an adenosine receptor-mediated process, but direct evidence for such a mechanism was not shown. The effective concentration of adenosine for inhibition of oocyte maturation is within the range of reported levels of adenosine in biological tissues and fluids.(ABSTRACT TRUNCATED AT 250 WORDS)

Adenine Nucleotides↗

Sensitive detection of the effects of reperfusion on myocardium by ultrasonic tissue characterization with integrated backscatter.

We have shown recently that tissue characterization of myocardium with ultrasound reflects changes associated with contractile function throughout the cardiac cycle. To determine whether ultrasonic tissue characterization can sensitively detect the impact of ischemic injury and reperfusion on contractile properties of the heart, we studied the time course of change of backscatter after 5, 20, and 60 min of coronary occlusion followed by reperfusion in 15 dogs. The time-averaged integrated backscatter (IB) and the amplitude and phase of cyclic variation of IB (phase relative to the left ventricular pressure waveform) were measured. A novel ultrasonic index of acute injury was identified, the phase-weighted amplitude of cyclic variation, and calculated by weighting the amplitude of cyclic variation of IB with respect to the phase. We hypothesized that backscatter variables would change dramatically after occlusion and that their restitution after reperfusion would sensitively reflect the extent and time course of reversibility of ischemic injury. After coronary occlusion, segmental wall thickening decreased from approximately 55% to 5% regardless of the duration of ischemia. Changes in backscatter associated with this decrease included an increase in time-averaged IB of approximately 5 dB, a 5 dB decrease in cyclic variation, an 80 degree phase shift, and a 7 dB decrease in phase-weighted amplitude. Wall thickening after reperfusion immediately after the 5, 20, or 60 min occlusions recovered to 45%, 27%, and 12% of baseline values, respectively. Within 3 hr it recovered to 53%, 44%, and 22%. Time-averaged IB recovered initially by 89%, 61%, and 44% (all p less than .05) and continued to recover subsequently although more slowly. Ultimate recovery was virtually complete. In contrast to the rapid recovery of time-averaged IB, phase-weighted amplitude recovered initially to only 72%, 41%, and -7% of baseline (all p less than .05) and manifested slower and incomplete recovery when ischemia had been present for 20 or 60 min. After reperfusion, the time course of both cyclic variation and phase were reflected by changes in the phase-weighted amplitude. The backscatter variables assessed appear to sensitively delineate the duration, time course of recovery, and reversibility of ischemic injury in response to reperfusion. The results suggest that early recovery of time-averaged IB corresponds in part to the restoration of tissue ultrastructural integrity.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Naturally occurring melioidosis in a colonized rhesus monkey (Macaca mulatta).

An aged wild-caught male rhesus monkey (Macaca mulatta), maintained in a research facility for 10 years, developed bilateral pelvic limb paralysis without other signs of disease. Unresponsive to therapy, the monkey was killed and necropsied. Chronic inflammation with osteolysis of thoracic vertebrae 10-13 was observed. Pseudomonas pseudomallei was cultured and identified from cerebrospinal fluid obtained at the site of the thoracic lesion. This Gram-negative bacterium can cause infection in animals and man and may remain latent for years before the appearance of clinical signs.

Animals↗

Hepatitis in an adult with rubella.

Rubella accompanied by serum aminotransferase elevations occurred in a 24-year-old woman. Although not generally recognized, hepatic involvement in adult rubella was the probable cause of her liver function test abnormalities. Sporadic hepatitis labeled as non-A, non-B may result from infection by common viruses such as rubella.

Adult↗

The dependence of myocardial ultrasonic integrated backscatter on contractile performance.

We have recently shown that the cardiac cycle-dependent variation in myocardial ultrasonic integrated backscatter is blunted with regional ischemia in dogs. To determine if global and intramural regional myocardial contractile performance can be quantified by integrated backscatter, we analyzed ultrasonic responses after induction of increased and decreased contractility in five dogs. A recently developed analog data-acquisition system for measuring integrated backscatter in real time was used to sample radiofrequency signals gated from subepicardial or subendocardial regions. Base-line recordings of integrated backscatter, left ventricular pressure, left ventricular dP/dt, and wall thickness were made at 12 left ventricular sites for both intramural regions. Contractility was modified subsequently by either paired pacing or propranolol to produce significantly elevated or depressed values for maximum left ventricular dP/dt compared with baseline (1083 +/- 289 to 3001 +/- 570 mm Hg/sec; p less than .01 for all). The amplitude of the cyclic variation of integrated backscatter was 50% greater (arithmetically) in subendocardial than in subepicardial regions for all treatments (7.6 +/- 0.3 vs 6.0 +/- 0.5 dB, p less than .001). The maximum rate of change in integrated backscatter waveforms during isovolumetric contraction was faster with paired pacing and slower with propranolol than at baseline for all regions (56 +/- 6 to 74 +/- 6 to 82 +/- 5 dB/sec, p less than .005). The maximum rate of change in integrated backscatter also was greater in subendocardial than subepicardial regions (p less than .001). Thus, both regional and global differences in myocardial contractile performance are manifest quantitatively in integrated backscatter waveforms. We propose that the physiologic determinants of these differences may depend on regional and global variations in myofibril elastic characteristics.

Animals↗

Effects of coronary artery occlusion and reperfusion on cardiac cycle-dependent variation of myocardial ultrasonic backscatter.

We have recently reported a systematic variation in integrated ultrasonic backscatter throughout the cardiac cycle in canine hearts. This study was performed to determine whether the pattern of such variation is modified systematically by ischemia. Measurements of integrated ultrasonic backscatter in selected regions of normal, ischemic, and reperfused hearts were compared in view of known differences in systolic function of myocardium in each of these regions. Integrated ultrasonic backscatter (3-7 MHz) gated to the first derivative of left ventricular pressure was measured at the apex, midwall, and base in 10 dogs and at the apex before and during transient ischemia and reperfusion in four dogs. Quantitative integrated ultrasonic backscatter was referenced to a steel reflector. Cyclic variation of integrated ultrasonic backscatter was greatest at the apex [peak to trough variation 5.5 +/- 0.9 dB (mean +/- SE)] with the maximum near end diastole (-52.9 +/- 0.9 dB) and minimum near end systole (-58.4 +/- 1.0 dB). Variation at the apex (5.5 +/- 0.9 dB) and the midwall (4.3 +/- 0.8 dB) was greater than at the base (0.5 +/- 1.0 dB) (P less than 0.01 for either region compared with base). Left anterior descending coronary occlusion for 10 minutes in four of 10 dogs reduced variation at the apex to 0.4 +/- 1.5 dB (P less than 0.02 compared with preocclusion). Reperfusion for 2 hours restored apical cyclic variation to 3.9 +/- 1.7 dB, i.e., to values not significantly different from those before occlusion.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

A relationship between ultrasonic integrated backscatter and myocardial contractile function.

We have shown previously that the physiologic, mechanical cardiac cycle is associated with a parallel, cardiac cycle-dependent variation of integrated backscatter (IB). However, the mechanisms responsible are not known. The mathematical and physiological considerations explored in the present study suggest that the relationship between backscatter and myocardial contractile function reflects cyclic alterations in myofibrillar elastic parameters, with the juxtaposition of intracellular and extracellular elastic elements that have different intrinsic acoustic impedances providing an appropriately sized scattering interface at the cellular level. Cardiac cycle-dependent changes in the degree of local acoustic impedance mismatch therefore may elicit concomitant changes in backscatter. Because acoustic impedance is determined partly by elastic modulus, changes in local elastic moduli resulting from the non-Hookian behavior of myocardial elastic elements exposed to stretch may alter the extent of impedance mismatch. When cardiac cell mechanical behavior is represented by a three-component Maxwell-type model of muscle mechanics, the systolic decrease in IB that we have observed experimentally is predicted. Our prior observations of regional intramural differences in IB and the dependence of IB on global contractile function are accounted for as well. When the model is tested experimentally by assessing its ability to predict the regional and global behavior of backscatter in response to passive left ventricular distention, good concordance is observed.

Animals↗

Amino acid transport in mouse blastocyst compartments.

Properties of uptake and exchange of L-methionine associated with the compartments of the blastocyst were examined in the mouse embryo. The inner cell mass (ICM) was derived from the 102 h blastocyst by immunosurgical procedures and studied in isolation. The ICM possessed strong exchange and efflux transport character. The Vmax of transport was about one-fifth that of the intact blastocyst, but the Km (660 microM) was about 30 times as high as that of the intact blastocyst. Consequently, at a concentration of about 100 microM-methionine, the the velocity of uptake into the ICM is about one-thirtieth of that into the intact blastocyst. Uptake of amino acid into the intact blastocyst was resolved into cellular and cavity components by mechanically collapsing the blastocyst following uptake of radiolabelled methionine. By this method, it was found that about 70% of the label accumulated by the blastocyst was in the cavity.

Amino Acids↗