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Biomedical subjects

D Manor

Publications and source records attributed to D Manor.

At least 55 records · Page 3Linked to original sources
PubMed1994

Discrete activation of transduction pathways associated with acetylcholine m1 receptor by several muscarinic ligands.

Activation of transfected muscarinic m1 acetylcholine receptors (m1AChR) has been linked to several signal transduction pathways which include phosphoinositide hydrolysis, arachidonic acid release and cAMP accumulation. In Chinese hamster ovary cells stably transfected with the rat m1AChR gene, carbachol elicited all three responses with EC50 values of 2.6, 3.8 and 76 microM, respectively. However, pilocarpine and the selective muscarinic agonist AF102B activated phosphoinositide hydrolysis (by 94 and 27% vs. carbachol, respectively), while antagonizing carbachol-mediated cAMP accumulation. Carbachol also activated (by 4-fold) adenylyl cyclase in membranes prepared from these cells, indicating independence of this signal from intracellular mediators. Moreover, carbachol and AF102B similarly elevated cytosolic Ca2+ in intact m1AChR-transfected cells. The ligand-selective cAMP accumulation, its independence from Ca2+ and the carbachol-activated adenylyl cyclase in membranes suggest that it represents an independent m1AChR-mediated signal, unrelated to phosphoinositide hydrolysis. Selective muscarinic ligands such as AF102B may independently activate distinct signalling pathways, which may be important for designing cholinergic replacement therapy for treating Alzheimer's disease.

Adenylyl Cyclases↗
PubMed1994

Hydrogen bond interactions of G proteins with the guanine ring moiety of guanine nucleotides.

We have utilized Raman difference spectroscopy to investigate hydrogen bonding interactions of the guanine moiety in guanine nucleotides with the binding site of two G proteins, EF-Tu (elongation factor Tu from Escherichia coli) and the c-Harvey ras protein, p21 (the gene product of the human c-H-ras proto-oncogene). Raman spectra of proteins complexed with GDP (guanosine 5' diphosphate), IDP (inosine 5' diphosphate), 6-thio-GDP, and 6-18O-GDP were measured, and the various difference spectra were determined. These were compared to the difference spectra obtained in solution, revealing vibrational features of the nucleotide that are altered upon binding. Specifically, we observed significant frequency shifts in the vibrational modes associated with the 6-keto and 2-amino positions of the guanine group of GDP and IDP that result from hydrogen bonding interactions between these groups and the two proteins. These shifts are interpreted as being proportional to the local energy of interaction (delta H) between the two groups and protein residues at the nucleotide binding site. Consistent with the tight binding between the nucleotides and the two proteins, the shifts indicate that the enthalpic interactions are stronger between these two polar groups and protein than with water. In general, the spectral shifts provide a rationale for the stronger binding of GDP and IDP with p21 compared to EF-Tu. Despite the structural similarity of the binding sites of EF-Tu and p21, the strengths of the observed hydrogen bonds at the 6-keto and 2-amino positions vary substantially, by up to a factor of 2.(ABSTRACT TRUNCATED AT 250 WORDS)

Binding Sites↗
PubMed1994

Modulation of small conductance calcium-activated potassium channels in C6 glioma cells.

Using the patch clamp technique, we have characterized a small conductance, calcium-activated potassium (SK) channel in the C6 glioma cell line. Elevation of cytosolic Ca2+ concentration ([Ca2+]i) by applications of serotonin or ionomycin induced bursts of channel opening recorded in the cell-attached configuration. These channels underlie the serotonin-induced, [Ca2+]i-activated whole-cell K+ conductance described previously. [Ca2+]i directly activated SK channels in inside-out patches with a biphasic concentration dependence. Submicromolar [Ca2+]i induced bursts of channel openings with a unitary conductance of about 25 pS, similar to that of the serotonin-induced channels. Supramicromolar [Ca2+]i caused prolonged openings with a unitary conductance of about 35 pS, resulting in a pronounced increase of the average current in patches exposed to [Ca2+]i above 100 microM. The two modes of opening reflect the activity of the same SK channel. The channel conductance depended on external K+ concentration with KD of 5 mM. The channel was slightly permeable to cations other than K+, with a permeability ratio for K+:Ca2+:Na+ of 1:0.040:0.030, respectively. ATP was required to maintain channel activity in outside-out patches but was not essential in inside-out patches. The modulations of SK channels in C6 cells by components in their microenvironment may be related to the role of glial cells in controlling the extracellular milieu in the CNS.

Adenosine Triphosphate↗
PubMed1994

Analysis of coronary circulation under ischaemic conditions.

Coronary flow patterns and pressure/flow relationships in coronary vessels with arterial stenoses are examined by using a model that combines the flow in the epicardial arterial tree with the intramyocardial perfusion. By using appropriate resistive elements, the model allows for the autoregulation of the vascular bed and for the development of coronary collaterals. Arterial flow predictions are compared to canine data. Coronary stenosis is simulated by a local pressure drop caused by a combination of viscous and inertial forces; stenosis with a constant cross-sectional area is compared to a dynamic stenosis in which the cross-sectional area is a function of the instantaneous transmural pressure. Simulation results predict that the normal phasic flow patterns in the epicardial arteries are unaffected up to 73% reduction in cross-sectional area, while the average flow remains unchanged up to 90% area reduction. At the critical level of 90% rigid stenosis, the autoregulation is saturated and the phasic nature of the arterial flow is severely damped. Dynamic stenoses demonstrate hysteresis loops of the instantaneous pressure/flow relationship. Theoretical predictions of local and global values are in excellent agreement with experimental measurements, indicating that the proposed approach can be used to realistically describe the coronary flow in the ischemic heart.

Animals↗
PubMed1994

Analysis of flow in coronary epicardial arterial tree and intramyocardial circulation.

A mathematical model combining the coronary flow in the epicardial arterial tree and the intramyocardial circulation is presented. The epicardial arterial tree is represented by a resistive capacitive network based on its realistic anatomy. The intramyocardial flow is affected by the pump action of the contracting myocardium through the extravascular compressive pressure (ECP), which, in turn, affects the dynamic resistance and compliance changes based on the relationship between the transmural pressure and the cross-sectional area of a vessel. The model accounts for the autoregulatory mechanism of the intramyocardial compartments (arteriolar, microvascular and venular) and is structured according to the epicardial coronary anatomy. Realistic coronary epicardial arterial flow patterns are obtained, which compare well to experimentally measured data in six dogs under basal conditions and during reactive hyperemic response. Simulations of the average transmural flow in the three intramyocardial vascular compartments show that the flow in the arterial side is predominantly diastolic, with a systolic retrograde component, and is dominantly systolic antegrade flow in the venular side, consistent with experimental data. Interestingly, the transmurally average microcirculatory flow is continuous, with very small change throughout the cardiac cycle, and is practically insensitive to changes in the model parameters. The model presents a quantitative tool that describes the dynamic patterns of coronary flow in relationship to muscular and extravascular parameters.

Animals↗
PubMed1994

Quantitative sorting of normal and abnormal coronary flow wave form shapes.

The normal phasic flow wave form in an epicardial coronary artery has a distinct characteristic shape, which reflects the interaction between the coronary tree, myocardial function and hemodynamic conditions. Since clinical measurements of phasic coronary wave forms are becoming available, determination of abnormal coronary flow wave forms is important. We suggest here an objective and automatic method to discriminate between normal and abnormal flow wave forms based on the Karhunen-Loève Transform (KLT), and experimentally tested it. The normal flow domain was represented by the resting flow waves measured in the left anterior descending arteries in 31 anesthetized dogs. The abnormal flow conditions, imposed and tested experimentally, were varying stenosis severity and severely reduced left ventricular pressure. In addition, the effects of reactive hyperemia on the shape of the flow were examined. The sorting index was based on the mean-square error (MSE) calculated for each flow signal based on a truncated KLT expansion. The results show excellent discrimination between the normal and the abnormal groups. During reactive hyperemia, however, MSE did not change significantly. These results indicate that the shape of abnormal coronary flow wave forms can be identified and discriminated from normal wave forms.

Animals↗
PubMed1994

In-vivo study of the mechanical properties of epicardial coronary arteries.

This study proposes a method to examine the in-vivo pressure-volume (P-V) relationship of an epicardial coronary artery. With the proximal left anterior descending (LAD) artery in a dog occluded, the distal LAD flow oscillates around zero. Integration of the flow with respect to time yields the intraluminal arterial segment volume changes in the region between the site of occlusion and the flow probe. The distal LAD pressure is measured via a diagonal branch. The segmental P-V relationship exhibits a hysteresis loop. The area bound by the hysteresis loop corresponds to the energy loss on the arterial wall during one cycle (12.63 +/- 8.25 [erg.cm-1], n = 7), while the dynamic arterial compliance is calculated based on the ratio of the volume to pressure excursions (1.13 +/- 0.73 [ml.mmHg-1.cm-1.10(-4)]). These results represent first estimates of the in-vivo mechanical properties of the coronary arterial wall based on P-V measurements.

Animals↗
PubMed1994

Pressure-flow characteristics of the coronary collaterals: a model study.

The pressure-flow relationship of the coronary collaterals is investigated by using an electrical analog model that combines the coronary epicardial arteries with the nonlinear characteristics of the intramyocardial circulation. The study aims to examine some controversial issues concerning the collateral circulation, including the transmural distribution of the collaterals, the distensibility of the collateral vessels (whether rigid or complaint), the effects of microcirculatory embolization, the collateral zero-flow pressure, and the nonlinearity of the collateral pressure-flow relationship. The study is carried out by simulating and comparing two basic experimental set-ups in which a coronary artery is ligated and the retrograde flow serves as an index of collateral flow. In the first "free-flow" setup, flow is allowed to bleed retrogradely against atmospheric pressure while perfusion pressure to the rest of the coronary arteries is varied over a wide range. In the second "back-pressure" setup, the coronary perfusion pressure is maintained at the control levels while the back pressure to the retrograde flow in the excised artery is varied. According to the analysis, the nonlinear pressure-flow relationships depend heavily on the experimental setup and are a function of the distensibility of the collaterals, which are distributed mainly on the epicardial surface, and the nonlinear contraction characteristics of the myocardium. The measured retrograde flow tends to underestimate the total collateral flow for the back-pressure setup because of antegrade flow escape.

Animals↗
PubMed1994

Reduced collateral perfusion is a direct consequence of elevated right atrial pressure.

In the present study we determined quantitatively the effects of increased right atrial pressure (RAP) on coronary and collateral flows. In an isolated, blood-perfused, maximally vasodilated dog heart preparation in which the left ventricle was vented, we used the retrograde flow method to assess collateral flow. When RAP was elevated from 5 +/- 1 (control) to 13 +/- 1 and 23 +/- 1 mmHg, retrograde flow from the left circumflex coronary artery (which was open to atmospheric pressure) increased 29 +/- 8 and 97 +/- 21% relative to control while left anterior descending flow decreased 5 +/- 1 and 14 +/- 2%, respectively (P < 0.01; n = 7). The increase in retrograde flow could be due to 1) an increase in collateral flow due to increased pressure at the origin of the collaterals or 2) the elevated RAP (venous outflow pressure), which forces the antegrade collateral flow component in the retrograde direction. To distinguish between these possibilities we embolized the circumflex with 30-microns spheres to eliminate the antegrade flow component. After embolization there was no significant change in retrograde flow with elevated RAP, indicating that the second supposition was correct. We conclude that increased RAP 1) results in a reduction of flow to the collateral-dependent myocardium and 2) reduces perfusion of the unoccluded coronary vessel. Furthermore, we found that under conditions of varying venous outflow pressure, retrograde flow may not serve as a reliable index of collateral flow.

Animals↗
PubMed1993

The interactions of retinoids with retinol-binding protein. A resonance Raman spectroscopic study.

We have measured the pre-resonance Raman spectrum of retinal, retinoic acid and retinol in dilute CCl4 solutions and when bound to the bovine-serum retinol-binding protein. The comparison reveals that the binding interaction does not involve any specific interactions of the head group and/or the polyene chain with a particular protein residue. The data indicate hydrogen bonding of bound retinal's head-group oxygen to water, as well as some torsional angle change of its polyene chain upon binding.

Ethylenes↗
PubMed1993

Myocardial mechanics and coronary flow dynamics.

The interaction between cardiac mechanics and coronary flow is highlighted here. Left ventricular (LV) structure and geometry are related to coronary flow dynamics and used in the analysis of experimental coronary flow data. The important role of the collagen mesh in the generation of the intramyocardial pressure (IMP), the pressure in the interstitial fluid, at a wide range of loading conditions is emphasized. The calculated IMP, based on a structural model of the LV myocardium, can explain most of the observed coronary compression characteristics under a variety of loading and contractility conditions. A more general compression function, the extravascular compressive pressure (ECP), is suggested to define coronary compression and is presented here based on the dynamics of the coronary inflow under constant perfusion conditions. Coronary compression is shown to be affected by fluid transport and the bi-directional coupling of coronary hemodynamics and IMP dynamics.

Animals↗
PubMed1992

[Diagnosis of acute appendicitis by transvaginal sonography].

Acute appendicitis is the most common indication for laparotomy. Nevertheless, diagnostic specificity is low and rates of false positive and false negative diagnoses are high, since there is no direct technique for visualizing the appendix. The ultrasonic appearance of appendicitis has recently been described. We present the first case of acute appendicitis identified via high frequency transvaginal sonography, in a 26-year-old woman. Because of the proximity to pelvic organs, transvaginal sonography may improve the diagnosis rate of acute appendicitis, especially in young women in whom the correct diagnosis is more difficult.

Acute Disease↗
PubMed1992

Calcium dependence of serotonin-evoked conductance in C6 glioma cells.

Whole-cell membrane currents and imaging of intracellular calcium concentrations ([Ca2+]i) were used to investigate the role of calcium in a response to serotonin of C6 glioma cells. Activation of a high-affinity serotonin receptor induced a transient rise in calcium concentration in these cells and activated a predominantly potassium conductance, with a small chloride component. Perfusion of the cytoplasm with an internal solution containing high calcium concentration induced similar but prolonged increase of membrane conductance. The responsiveness of C6 cells to serotonin was negatively correlated with the concentration of the unbound calcium chelator BAPTA when BAPTA-buffered calcium-containing intracellular solutions were used. Responses to serotonin persisted in the absence of external calcium, decreased gradually, and then recovered partially after replenishment of extracellular calcium. These findings substantiate the direct role of intracellular calcium in mediating the serotonin response, and indicate that serotonin-induced release of calcium from intracellular stores is sufficient for the activation of conductance in the C6 glioma cell line.

Brain Neoplasms↗
PubMed1992

Confocal microscopic imaging of [Ca2+]i in cultured rat hippocampal neurons following exposure to N-methyl-D-aspartate.

1. The confocal laser scanning microscope (CLSM) was used in conjunction with the calcium indicator dye Fluo-3 to record changes in free intracellular calcium concentration ([Ca2+]i) in cultured hippocampal neurons in response to superfusion of N-methyl-D-aspartate (NMDA). 2. NMDA caused a rapid rise in [Ca2+]i in all parts of the neuron. The rise in [Ca2+]i was dependent on activation of an NMDA receptor, was enhanced by the removal of Mg2+ and addition of glycine to the superfusion medium, and was dependent on normal [Ca2+]o. 3. The rise of [Ca2+]i was seen first near the membrane. A wave of elevated [Ca2+]i moved centripetally at a rate of 117 microns/s. 4. Dantrolene pre-incubation caused a significant reduction in the efficacy of the NMDA-induced rise in [Ca2+]i, indicating that at least part of the rise is caused by intracellular release of calcium. 5. The replacement of calcium by barium caused a reduction in the response to NMDA, but a significant response was still present in these cells, supporting the assumption that NMDA causes release of calcium from intracellular stores. 6. The removal of sodium from the superfusion medium prolonged the [Ca2+]i rise in response to NMDA indicating that the Na-Ca antiporter is instrumental in reducing [Ca2+]i. 7. These studies demonstrate the multiplicity of regulating mechanisms of [Ca2+]i following activation of NMDA receptors.

Animals↗
PubMed1992

Is cord blood erythropoietin a marker of intrapartum hypoxia?

A sensitive assay was used to compare the biological activity of cord serum erythropoietin in two groups of infants born with or without labor-induced hypoxia. The mean cord serum erythropoietin activity in 161 infants delivered after vaginal labor was 116 +/- 36 mU/mL, and was indistinguishable from that observed in 23 infants delivered by preplanned, elective cesarean section, 114 +/- 12 mU/mL (P = .75). The bioassay measured effective erythropoietin activity, including the contribution of potentiators in serum. These results indicate that duration and intensity of labor are insufficient to cause a significant increase in effective erythropoietin activity.

Cesarean Section↗
PubMed1991

An isotope edited classical Raman difference spectroscopic study of the interactions of guanine nucleotides with elongation factor Tu and H-ras p21.

We have measured the Raman spectrum of GDP bound to the elongation factor protein, EF-Tu, and the c-Harvey-ras protein, p21, two proteins of the guanine nucleotide binding family. In order to separate the Raman spectrum of the nucleotide from the much more intense protein spectrum, we investigate the feasibility of "tagging" the normal modes of the nucleotide by isotopic substitution, here by incoporating deuterium-labeled guanine at the C8 position into the active site. A difference spectrum between the labeled and unlabeled protein-nucleotide complex shows the changes in the Raman spectrum of the bound nucleotide that arise from the isotopic exchange. We find that surprisingly good Raman spectra of bound ligands can be obtained with this method and that the method can be easily generalized to other systems. The data show that the guanine amino group of the nucleotide interacts differently with both EF-Tu and p21 than it does with water, showing a change in hydrogen-bonding properties upon binding. On the other hand, no change in hydrogen bonding is observed at guanine's N7. The data strongly suggest that the conformation of the nucleotide when bound to EF-Tu and that p21 is the C2' endo pucker of the ribose ring and anti about the glycosidic bond. These results are compared to previous structural and chemical studies.

Amino Acid Sequence↗
PubMed1991

Ultrafast spectroscopy of the visual pigment rhodopsin.

We report on time-resolved absorption studies of the bovine visual pigment rhodopsin with subpicosecond resolution at room temperature. Our data show that bathorhodopsin, rhodopsin's early photoproduct, is photochemically formed in 3.0 +/- 0.7 ps. The data suggest that bathorhodopsin formation is kinetically preceded by two species along the rhodopsin-to-bathorhodopsin reaction coordinate. The first is identified with the vertically excited Franck-Condon state. This decays with an approximately 200-fs lifetime to an intermediate, which then decays to bathorhodopsin in 3.0 ps. We assign this intermediate to be an excited state transient near 90 degrees along the 11-12 torsional coordinate of rhodopsin's chromophore. Exchange of rhodopsin's exchangeable protons for deuterons does not affect the observed dynamics. These observations are both qualitatively and quantitatively consistent with molecular dynamics calculations, which model the rhodopsin to bathorhodopsin phototransition as a cis-trans isomerization along the 11-12 torsional coordinate of rhodopsin's chromophore.

Animals↗