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Microscopic susceptibility variation and transverse relaxation: theory and experiment.

Microscopic susceptibility variations invariably increase apparent transverse relaxation rates. In this paper, we present comparisons between Monte Carlo simulations and experiments with polystyrene microspheres to demonstrate that this enhanced relaxation can be explained quantitatively for both spin echo and gradient echo imaging experiments. The spheres used (1 to 30 microms), and degree of susceptibility variation (caused by 0-12 mM Dy-DTPA) covered a wide range of biologically relevant compartment sizes and contrast agent concentrations. These results show that several regimes of behavior exist, and that contrast dependence is quite different in these regimes. For a given susceptibility, delta chi, a small range of particle sizes show peak transverse relaxation. For the range of susceptibilities found in the first pass of a clinical IV contrast agent bolus, this size range is 5 to 10 microns, or roughly capillary sized compartments. In both our simulations and experiments, smaller spheres showed quadratic relaxation versus concentration curves, and larger particles showed sublinear behavior. For particles corresponding to the peak relaxivity, the relaxation-concentration curves were linear. In addition, we demonstrated that increasing the diffusion coefficient can increase, decrease, or, paradoxically, leave unaffected the apparent relaxation rate. The regime for which the diffusion coefficient is relatively unimportant corresponds to the region of peak relaxivity. By using the Bloch-Torrey equation to produce scaling rules, the specific Monte Carlo simulations were extended to more general cases. We use these scaling rules to demonstrate why we often find that susceptibility-induced relaxation rates vary approximately linearly with concentration of injected agent.

Animals↗

Integrated analysis of diffusion and relaxation of water in blood.

Diffusion and T2 relaxation of water both inside and outside red blood cells (RBCs) in human blood were investigated using a hybrid NMR pulse sequence to obtain a more quantitative understanding of the diffusion and relaxation behavior of water in paramagnetic-doped blood samples. The data were analyzed by both examining the relaxation properties of the system after each diffusion weighting and looking at the diffusion properties at each echo time. The results illustrate how diffusion-sensitizing gradients affect the T2 spectra of blood and how relaxation weighting changes the curvature of the diffusion curves, thereby demonstrating the close coupling between diffusion and T2 relaxation. A three-pool model, consisting of RBCs, plasma, and macromolecular protons, was used to model the data from the diffusion-relaxation hybrid experiments. The model was found to describe all the characteristic features of the experimental data well and was used to evaluate the approximations involved in the conventional analysis methods and elucidate the nature of the relaxing and diffusing components. Compared with the separate diffusion and relaxation experiments, the diffusion-relaxation hybrid experiments are less time-consuming, result in better parameter determinations, and may be useful in analyzing diffusion-T2 coupling in tissues with more complicated multiexponential T2 behavior.

Biological Transport↗

Hydroxyl radical--a mediator of acetylcholine-induced vascular relaxation.

It is well known that acetylcholine (Ach)-induced vasodilation is mediated through the release of the endothelium-derived relaxing factor (EDRF), nitric oxide (NO.). It has been suggested that NO interacts with superoxide anion (O2-) to generate peroxynitrite which at physiological pH gives rise to peroxynitrous acid than rapidly decomposes to hydroxyl radical (.OH) and nitrogen dioxide, .OH relaxes isolated aorta. It was hypothesized that Ach-induced vascular relaxation is mediated by .OH derived from interaction of NO and O2-. To test this hypothesis we investigated the effect of Ach on norepinephrine (NE)-precontracted isolated rabbit aortic preparations in the absence or presence of scavengers of O2- [superoxide dismutase (SOD)] and of .OH [dimethylthiourea (DMTU) or mannitol]. .OH and Ach produced relaxation of NE-precontracted preparations in a concentration-dependent manner. Relaxation produced by .OH generating system was prevented by mannitol, a .OH scavenger suggesting that relaxation is due to .OH. SOD. DMTU, mannitol or combination of SOD and DMTU markedly reduced the Ach-induced relaxation. Glyburide (an ATP-sensitive K+ channel blocker) was ineffective in blocking the Ach-induced remaining relaxation in the presence of SOD and DMTU. These results suggest that .OH formed from interaction of O2- and NO. is the mediator of Ach-induced vascular relaxation. Thus while EDRF is NO. its mechanism of action involves .OH.

Acetylcholine↗

Off-resonance rotating-frame spin-lattice relaxation of quadrupolar (spin-1) nuclei.

The validity of the formalism for the off-resonance rotating-frame spin-lattice relaxation experiment applicable to spin-1 quadrupolar nuclei was experimentally examined by considering two model systems, deuterated glycerol and deuterated benzene in castor oil, at different temperatures. When appropriately implemented, the deuterium off-resonance rotating-frame spin-lattice relaxation experiment provides spectral-intensity-ratio-dispersion data which agree remarkably well with those predicted by the theoretical formalism. The assumption of quadrupolar relaxation as the dominant relaxation mechanism, and rigid-rotor isotropic tumbling, permits the assessment of rotational diffusion behavior, i.e., the determination of a rotational correlation time, of a variety of molecular systems. With the inclusion of an additional relaxation measurement, T1 or T2, the 2H off-resonance rotating-frame spin-lattice relaxation experiment becomes a convenient method for the estimation of the 2H quadrupolar coupling constant, provided that a realistic reorientational motional model is assumed in the theoretical relaxation expressions used in the analysis. Because the motional window of the 2H off-resonance rotating-frame spin-lattice relaxation experiment includes intermediate molecular motion with correlation times as short as 1 or 2 ns, this experiment is appropriate for the investigation of the rotational diffusion behavior of deuterated molecules varying in size from moderately small to macromolecular. The 2H off-resonance rotating-frame spin-lattice relaxation experiment is applicable to in vitro and in vivo experimental situations.

Algorithms↗

Cross-correlated relaxation for the measurement of angles between tensorial interactions.

Theory,experimental aspects, and use in structure calculation of cross-correlated relaxation rates measured on zero- and double-quantum coherences in liquid state NMR are presented. The relative size of the interaction depends on the projection angle between the two tensorial interactions. The tensorial interaction can be either a dipolar interaction or a chemical shift anisotropy relaxation mechanism (CSA). Effects of additional sources of relaxation on the cross-correlated relaxation rates are analyzed. Also, an easy-to-use formalism is given to manipulate different cross-correlated relaxation interactions. The application addresses measurement of the backbone angle psi in a protein by measuring dipole((15)N-(1)H)-dipole((13)C(alpha)-(1)H(alpha)) and CSA((15)N)-dipole((13)C(alpha)-(1)H(alpha)) cross-correlated relaxation rates. It is shown that ambiguities due to the 3 cos(2)θ-1 dependence of one cross-correlated relaxation rate can be overcome by measuring additional cross-correlated relaxation rates. The use of cross-correlated relaxation rates is demonstrated in structure calculations.

Anisotropy↗

Characterization of dopamine-mediated relaxation in experimental vein bypass grafts.

BACKGROUND: Dopamine is an endogenous inotropic agent commonly used during coronary artery surgery and in the medical therapy of a revascularized patient. In this study the responses of intimal hyperplastic vein grafts to dopamine are examined. METHODS: The in vitro isometric tension responses to dopamine of common carotid jugular vein bypass grafts in New Zealand White rabbits were determined. The responses were compared to those obtained in the jugular vein and in the common carotid artery. Both endothelialized and denuded vessels were precontracted with prostaglandin F(2alpha) and the responses to dopamine were assessed. The contributions of nitric oxide and prostanoids to the response were also determined. RESULTS: Each vessel showed a biphasic dose response to dopamine with relaxation at low concentrations followed by contraction at high concentrations. Dopamine relaxation in the jugular vein was endothelial independent while in the carotid artery it was endothelial dependent and decreased. The sensitivity of both vessels was significantly greater than the vein graft (6.62 +/- 0.12; P < 0. 05); however, after endothelial denudation, the sensitivity of dopamine-mediated relaxation of the vein graft (8.91 +/- 0.09) was significantly enhanced. Preincubation with L-NMMA (to block NO synthesis) inhibited vein graft relaxation to dopamine and preincubation with indomethacin (to block cyclooxygenase activity) inhibited carotid artery relaxation to dopamine. Addition of phenoxybenzamine, a broad alpha-adrenergic antagonist, enhanced dopamine relaxation in the jugular vein and depressed the relaxation in the carotid artery. There was no effect on the dopamine response in the vein graft. Jugular vein and carotid artery responded to dopamine with cholera toxin-sensitive (Galpha(s)) responses. In contrast, dopamine relaxation in the vein graft was enhanced by inhibition of Galpha(s). CONCLUSION: Dopamine relaxation in vein grafts is mediated in part by NO but not by either prostanoids or alpha-adrenergic receptor activation. It is diminished compared to native vessels due to an endothelium-dependent, Galpha(s)-mediated pathway.

Adjuvants, Immunologic↗

Levcromakalim- and isoprenaline-induced relaxation of human isolated airways--role of the epithelium and of K+ channel activation.

In this study we have investigated the mechanism of action of levcromakalim and isoprenaline in human isolated airways with respect to the K+ channels they activate and the possibility that these smooth muscle relaxants activate K+ channels on the airway epithelium. Mechanical removal of the epithelial layer (mean percentage of epithelium present 20 +/- 3%, n = 20 tissues) did not affect the relaxation responses to levcromakalim or isoprenaline, either in terms of maximal relaxation or sensitivity. Whilst having no effect on isoprenaline-induced relaxation, studied from basal tone, the ATP-sensitive K+ channel blocker BRL 31660 (10, 30 and 50 microM) reduced relaxation responses induced (from basal tone) by levcromakalim from 74 +/- 6% (of the maximal response to isoprenaline) to 48 +/- 12% (n = 7), 9 +/- 9% (n = 4) and 0 (n = 4), respectively. Charybdotoxin, a blocker of high conductance Ca(2+)-activated K+ channels, at concentrations of 30 and 100 nM, had no effect on either levcromakalim- or or isoprenaline-induced relaxation responses and yet charybdotoxin was active at KCa channels in outside-out patches of hippocampal granule cells. Moreover, tetraethylammonium (10 mM) inhibited neither isoprenaline- nor levcromakalim-induced relaxation. This study has demonstrated that the relaxation responses elicited in human bronchus to isoprenaline and levcromakalim are likely to be the result of direct effects on the smooth muscle with no contribution from epithelial receptors or K+ channels. The actions of levcromakalim appear to be mediated only via activation of KATP channels. Further, we have made the important observation that, under the experimental conditions of our study, isoprenaline does not activate the KCa channel to produce relaxation in human bronchus.

Adult↗

Analysis of cromakalim-, pinacidil-, and nicorandil-induced relaxation of the 5-hydroxytryptamine precontracted rat isolated basilar artery.

The effects of the K+ channel activators cromakalim, pinacidil, and nicorandil were investigated in endothelium intact, 5-hydroxytryptamine (5-HT) precontracted rat isolated basilar artery. Cromakalim, pinacidil, and nicorandil produced concentration-dependent relaxation of rat isolated basilar artery precontracted with 5-HT with a rank order of potency of cromakalim greater than pinacidil greater than nicorandil. All compounds produced full or nearly full relaxation. The calculated Hill coefficients for cromakalim-, pinacidil-, and nicorandil-induced relaxation of 5-HT-precontracted rat isolated basilar artery were 2.20 +/- 0.36, 1.30 +/- 0.07, and 1.00 +/- 0.01, respectively. Under conditions of increased tone produced by 50 mmol/l KCl (which inhibits cromakalim-induced relaxation) pinacidil and nicorandil produced marked reversal of spasm, with pinacidil being more potent than nicorandil. In arteries precontracted with 5-HT, preincubation with glibenclamide (0.1-1 mumol/l) produced concentration-related inhibition of relaxation with calculated mean pA2 values (and slopes of Schild regression) +/- SEM of 6.84 +/- 0.20 (1.1 +/- 0.20) against cromakalim. 6.60 +/- 0.14 (0.95 +/- 0.23) against nicorandil, and 6.57 +/- 0.26 (1.04 +/- 0.18) against pinacidil. For cromakalim, pinacidil, and nicorandil the slopes of Schild regression were not significantly different from unity. Tolbutamide 10 mumol/l was without effect against the cromakalim-, pinacidil-, or nicorandil-induced relaxation. Tetraethylammonium (TEA; 1-10 mmol/l) produced noncompetitive inhibition of the cromakalim-induced relaxation, but appeared to produce competitive inhibition of the pinacidil- and nicorandil-induced relaxations. We conclude that cromakalim, pinacidil, and nicorandil produce relaxation of the 5-HT precontracted rat basilar artery by similar mechanisms to those identified in other peripheral vascular and visceral smooth muscle.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Differential antagonism by glibenclamide of the relaxant effects of cromakalim, pinacidil and nicorandil on canine large coronary arteries.

The relaxant mechanisms of action of cromakalim, pinacidil and nicorandil, potassium channel openers, on large epicardial coronary arteries were investigated in isolated canine left circumflex arteries contracted by 10(-7) mol/l U46619, a thromboxane A2 analogue, or addition of 25 mmol/l KCl in comparison with nitroglycerin. Cromakalim (3 x 10(-8)-3 x 10(-5) mol/l), pinacidil (10(-6)-10(-4) mol/l), nicorandil (3 x 10(-6)-10(-3) mol/l) and nitroglycerin (3 x 10(-8)-10(-5) mol/l) all produced a concentration-dependent relaxation in both U46619- or KCl-contracted arteries. At their maximum effects pinacidil, nicorandil and nitroglycerin produced full relaxation in arteries contracted by either means. In contrast, cromakalim produced about a 73% relaxation in KCl-contracted arteries, although it caused full relaxation in U46619-contracted ones. In the presence of glibenclamide the concentration-relaxation curves for cromakalim in U46619- or KCl-contracted arteries underwent rightward parallel shifts. Schild regression had a slope of 1.00 and yielded a pA2 of 7.47 for glibenclamide in U46619-contracted arteries, and corresponding values obtained in KCl-contracted arteries were 0.86 (not significantly different from unity) and 7.28. The concentration-relaxation curves for pinacidil in U46619-contracted arteries also underwent rightward parallel shifts in the presence of glibenclamide, however, Schild regression had a slope of 0.60. The concentration-relaxation curves for pinacidil in KCl-contracted arteries underwent rightward parallel shifts only to a limited extent in the presence of glibenclamide. The concentration-relaxation curves for nicorandil and nitroglycerin in U46619- or KCl-contracted arteries were not affected by glibenclamide in concentrations which antagonized cromakalim.(ABSTRACT TRUNCATED AT 250 WORDS)

15-Hydroxy-11 alpha,9 alpha-(epoxymethano)prosta-5↗

19F NMR relaxation studies on 5-fluorotryptophan- and tetradeutero-5-fluorotryptophan-labeled E. coli glucose/galactose receptor.

19F NMR relaxation studies have been carried out on a fluorotryptophan-labeled E. coli periplasmic glucose/galactose receptor (GGR). The protein was derived from E. coli grown on a medium containing a 50:50 mixture of 5-fluorotryptophan and [2,4,6,7-2H4]-5-fluorotryptophan. As a result of the large gamma-isotope shift, the two labels give rise to separate resonances, allowing relaxation contributions of the substituted indole protons to be selectively monitored. Spin-lattice relaxation rates were determined at field strengths of 11.75 T and 8.5 T, and the results were analyzed using a model-free formalism. In order to evaluate the contributions of chemical shift anisotropy to the observed relaxation parameters, solid-state NMR studies were performed on [2,4,6,7-2H4]-5-fluorotryptophan. Analysis of the observed 19F powder pattern lineshape resulted in anisotropy and asymmetry parameters of delta sigma = -93.5 ppm and eta = 0.24. Theoretical analyses of the relaxation parameters are consistent with internal motion of the fluorotryptophan residues characterized by order parameters S2 of approximately 1, and by correlation times for internal motion approximately 10(-11)s. Simultaneous least squares fitting of the spin-lattice relaxation and line-width data with tau i set at 10 ps yielded a molecular correlation time of 20 ns for the glucose-complexed GGR, and a mean order parameter S2 = 0.89 for fluorotryptophan residues 183, 127, 133, and 195. By contrast, the calculated order parameter for FTrp284, located on the surface of the protein, was 0.77. Significant differences among the spin-lattice relaxation rates of the five fluorotryptophan residues of glucose-complexed GGR were also observed, with the order of relaxation rates given by: R1F183 > R1F127 approximately R1F133 approximately R1F195 > R1F284. Although such differences may reflect motional variations among these residues, the effects are largely predicted by differences in the distribution of nearby hydrogen nuclei, derived from crystal structure data. In the absence of glucose, spin-lattice relaxation rates for fluorotryptophan residues 183, 127, 133, and 195 were found to decrease by a mean of 13%, while the value for residue 284 exhibits an increase of similar magnitude relative to the liganded molecule. These changes are interpreted in terms of a slower overall correlation time for molecular motion, as well as a change in the internal mobility of FTrp284, located in the hinge region of the receptor.

Anisotropy↗

[Biomechanics of human tendons: connection between stress relaxation and stress recovery (author's transl)].

108 tendons of the m. extensor hallucis longus were examined with a tensile testing machine within 36 h after death. The specimen were kept at a resting length of 20 mm. After the "steady state" was reached by cyclic loading, the tendons were stretched up to a maximum load of 18 kp, then deloaded to a certain level and after that the elongation was kept constant. At high loading level the tension of the tendon decreases with time (relaxation). At medium and low loading level the tension increases slightly (mechanical recovery). Between that two regions there is a certain load, where the tension will not change with time (isorheological point). The position of the isorheological point depends on the velocity of the elongation. At low velocity (2 mm/min) the isorheological point is situated at 70%, at high velocity (12 mm/min) at 60% of the maximum load. One will find the maximum relaxation, when no deloading occurs. The mechanical recovery, however, has its maximum at 5--25% of the maximum load. But when the tendon is totally deloaded, there seems to occur no recovery. The maximum relaxation is 5 to 6 times larger than the maximum recovery. Supposingly the relaxation- and recovery-processses will happen at the same time but with different intensity depending on the loading level. At least the relaxation-process consists of different relaxation components with different relaxation times. This will explain the phenomenon of a "secondary relaxation": After a long time of registration the recovery will turn into a slight relaxation.

Adolescent↗

Gradient of pressure and time between proximal anal canal and high-pressure zone during internal anal sphincter relaxation. Its role in the fecal continence mechanism.

PURPOSE: The normal response to rectal distention is a relaxation of the proximal anal canal (PAC). We hypothesized that this mechanism would require a gradient of pressure and time to preserve continence. METHODS: Sixteen volunteers (10 male), mean age, 41.5 (range, 24-60) years, were studied using an eight port axial catheter with a compliant balloon at its tip. Relaxation was induced by a small volume of rectal distention (15-30 ml of air) and was recorded until recovery of resting anal pressure (RAP). Duration of relaxation was measured until recovery of RAP. Amplitude of relaxation was determined between RAP before rectal distention (RAP-BR) and pressure at the point of maximum relaxation (RAP-PMR). Gradient of pressure was determined by comparing RAP-PMR in the high-pressure zone (HPZ) and PAC. Contraction in the distal anal canal was interpreted as external anal sphincter contraction (EASC) and was compared with RAP-PMR in the HPZ. RESULTS: Relaxation was significantly greater in PAC than in HPZ (50 vs. 36 percent; P = 0.001). RAP-PMR was significantly higher in HPZ than in PAC (30.7 vs. 12.6 mmHg; P = 0.001). EASC was observed in six patients and did not show significant difference with RAP-PMR in HPZ (39.7 vs. 36.3 mmHg; not significant). Relaxation began at the same time in all levels but lasted significantly longer in PAC compared with HPZ (13.5 vs. 9.4 sec; P = 0.003). CONCLUSION: Anal relaxation induced by small volume rectal distention involves a gradient in the pressure and time of relaxation between PAC and the HPZ.

Adult↗

Vascular effects of Siberian ginseng (Eleutherococcus senticosus): endothelium-dependent NO- and EDHF-mediated relaxation depending on vessel size.

Siberian ginseng (SG) has been widely and historically consumed as a health food product for the improvement of self well-being, but whether vascular relaxation may contribute to such a therapeutic health effect has not been studied. We therefore investigated the vasorelaxant effect of the aqueous extract of the roots of SG (Eleutherococcus senticosus Maxim) using several in vitro vascular rings prepared from dog carotid artery, rat aorta and rat mesenteric artery. SG extract (0.04-0.8 mg/ml) caused concentration-dependent relaxation in dog carotid arterial rings pre-contracted with 100 microM phenylephrine (PE), and the relaxation was primarily endothelium-dependent. Treatment with 100 microM L-NOARG (a nitric oxide synthase inhibitor) either prevented or totally reverted SG-induced relaxation, suggesting that the endothelium-dependent relaxation was mediated by NO. Similar endothelium-dependent vascular relaxant responses were also obtained with rat aortic and mesenteric arterial rings, except that it occurred over a relatively higher concentration range of SG (0.5-2.0 mg/ml). When tested in the presence of 300 microM L-NAME, the vasorelaxant effect of SG was inhibited totally in rat aorta but only partially in rat mesenteric artery. The relaxation to SG that was insensitive to L-NAME in rat mesenteric arterial rings was eliminated when the rings (both proximal and distal ends) were pre-treated with a combination of 300 microM L-NAME and 15 mM KCl indicating the involvement of endothelium-derived hyperpolarizing factor (EDHF). This vasorelaxant response of the SG extract was inhibited partially by atropine (1 microM), completely by TEA (5 mM), but not by indomethacin (1 microM) or propranolol (10 microM). SG up to 2 mg/ml had no effect on KCl-induced contraction in any of the vascular rings studied. When compared with carbachol-induced (CCh) relaxation, SG resembles CCh in that the sensitivity to L-NAME inhibition is dependent on vascular size, i.e. aorta >proximal end of mesenteric artery >distal end of mesenteric artery. However, SG exhibited different potencies to relaxation while CCh showed similar potency (EC(50) of about 0.2 microM) in all three vascular segments. In conclusion, we have demonstrated that the vascular effect of SG is endothelium-dependent and mediated by NO and/or EDHF depending on the vessel size. Other vasorelaxation pathways, such as inhibition of K(+)-channels and activation of muscarinic receptors, may also be involved.

Animals↗

From metmyoglobin to deoxy myoglobin: relaxations of an intermediate state.

Metmyoglobin has been reduced at low temperature (below 100 K) using x-rays or by excitation of tris(2,2,bipyridine)ruthenium(II) chloride with visible light. Upon reduction, an intermediate state is formed where the structure of the protein is very similar to that of metmyoglobin with the water molecule still bound to the heme iron, but the iron is II low spin. The nature of the intermediate state has been investigated with optical spectroscopy. The Q(O) and Q(V) bands of the intermediate state are split, suggesting that the protoporphyrin is distorted. The intermediate state undergoes a relaxation observed by a shifting of the Soret band at temperatures above 80 K. Above 140 K, the protein begins to relax to the deoxy conformation. The relaxation kinetics of the protein have been monitored optically as a function of time and temperature from minutes to several hours and from 150 K to 190 K. By measuring the entire visible spectrum, we are able to distinguish between electron transfer processes and the protein relaxation from the intermediate state to deoxy myoglobin. The relaxation has been measured in both horse myoglobin and sperm whale myoglobin with the relaxation occurring on faster time scales in horse myoglobin. Both the reduction kinetics and the relaxation show non-exponential behavior. The reduction kinetics can be fit well to a stretched exponential. The structural relaxation from the intermediate state to the deoxy conformation shows a more complex, dynamical behavior and the reaction is most likely affected by the relaxation of the protein within the intermediate state.

2,2'-Dipyridyl↗

Microscopic photometric quantification of stiffness and relaxation time of red blood cells in a flow chamber.

The Microscopic Photometric Monolayer Technique provides a tool to measure red blood cell (RBC) stiffness (resistance to elongation) and relaxation time. It combines many of the advantages of flow channel studies of point-attached RBCs with the simplicity, sensitivity and accuracy of photometric light transmission measurement. This technique allows the study of the effects of physicochemical factors on the elongation and relaxation time of the same cells within an average of four to five thousand cells adhered as a monolayer to glass. Further, the time course of physicochemical effects on cell membrane and wash-in/wash-out kinetics of interactions can be followed. An automated version of this technique was developed. A dense monolayer of point-attached RBCs was prepared at the bottom of a flow-chamber. A steady-state flow, with stepwise increases of flow rate, induced the RBC elongation. The light transmission perpendicular through the monolayer plane was measured photometrically. Photomicrographs compared with photometric results showed that the flow-induced bending and curvature change of RBC membrane was associated with the increase of light transmission. There was a linear correlation between the photometric index of elongation and the elongation taken from photomicrographs for shear stresses up to 0.75 Pa. A stiffness parameter, S (in Pa), was defined as the ratio of shear stress and elongation at a shear stress of 0.25 Pa. Following a sudden flow stoppage, the RBCs returned to their resting shape and the RBC relaxation time was measured. The stiffness-relaxation time product, V (in mPas), was calculated to provide an estimate of viscosity. Diamide treatment, known to stiffen RBCs, did result in dose-dependent decreases of elongation and relaxation time. With increasing temperature, the relaxation time decreased at a rate of -2.96 ms/K; the stiffness increased significantly at a rate of 0.0038 Pa/K, and the stiffness-relaxation time product decreased with -2.95 mPas/K, reflecting an inverse relationship between RBC viscosity and temperature. Using the automated version of this technique (Elias-c-) to test RBCs of 36 healthy subjects, we found the inter-individual coefficients of variation to be 8.6% for stiffness, 7.9% for relaxation time and 12.4% for stiffness-relaxation time product.

Adult↗

The effect of diastolic vibration on the relaxation of rat papillary muscle.

OBJECTIVE: It has been demonstrated that the application of an external mechanical vibration during the ventricular relaxation phase of the heart (diastolic vibration) decreases the time needed for myocardial relaxation. The objective of this study is to see whether this vibration-induced decrease in relaxation time is an intrinsic property of the contractile proteins. We hypothesize that this decrease in duration of the late systolic phase by diastolic vibration is likely to be due to a forced detachment of crossbridges from the actin filament. This would then result in a decrease in the relaxation time of myocardial tissue. METHODS: A controlled vibration of variable amplitude and frequency was applied to isolated twitching rat papillary muscles. Vibration was initiated from directly after peak tension development and ended when tension had returned to baseline. Effects of applied vibration were expressed as changes in the time interval from 90% rising- to 50% falling-force level, which was termed the 'late systolic phase'. RESULTS: The data showed that in general diastolic vibration decreased the duration of this late systolic phase. A vibration of 1.0% of Lmax at 50 Hz in the late systolic phase shortened this period by 27 ms (20-30%) on average. At increasing amplitude of vibration the increase in the rate of relaxation was even more pronounced. Shortening of this duration was slightly less at frequencies of vibration below 40 Hz than at higher frequencies of vibration. Changes in the resting muscle length did not result in significant changes in shortening of the studied relaxation period. CONCLUSION: The results show that relaxation is accelerated during application of an external vibration during the period of tension fall of an isolated cardiac muscle preparation. Therefore, we can conclude that the acceleration of relaxation due to vibration is primarily due to properties of the myocytes themselves rather than to the complex geometric structure of the heart. Tension in the relaxation phase was reduced during diastolic vibration, which suggests that the number of crossbridges bound to the actin filament was reduced by the length perturbation.

Animals↗

Correlation of lysophosphatidylcholine-induced vs spontaneous relaxation to cyclic GMP levels in rabbit thoracic aorta.

We demonstrated previously that 10(-5)M micellar solution of lysophosphatidylcholine (LPC) produced relaxation of rabbit thoracic aorta in a dose-dependent manner. It was also noticed that histamine (HA)-precontracted strips relaxed spontaneously in the absence of LPC. We now measured the cyclic GMP changes during these two types of relaxation. Results indicate that while LPC-induced relaxation was mediated through cyclic GMP, spontaneous relaxation was not. The vasodilating effect of LPC was not due to its interference with cellular viability by solubilizing the membrane, because repeated additions of LPC produced identical degrees of relaxation and after three consecutive additions of LPC, the strips continued to relax to the same degree with acetylcholine (Ach) demonstrating the functional integrity of the endothelium. LPC/Cholesterol dispersions (1:0.5 mole percent) relaxed the strips to the same extent as the micellar solution of LPC, while 1:1 mole percent did not. The results stress the role of cyclic GMP in LPC-induced relaxation. They further suggest that LPC/Cholesterol ratio may determine the availability of LPC and regulate LPC-mediated processes.

Animals↗

Investigation of the interrelationship between coagulation and thrombin-induced EDRF-dependent relaxation in dog coronary artery.

Little is known about the possible interrelationships between thrombin-induced EDRF-dependent vascular relaxation and coagulant activity. We have now studied the effects of the anticoagulant zymogen protein C, on EDRF-dependent relaxation in isolated canine coronary arteries. Low concentrations of activated protein C (0.1-30 ng/ml) had no significant effect, but higher concentrations caused relaxation (Emax -39.2 +/- 7.2%; 100-1000 ng/ml). To determine whether relaxation was dependent on coagulation complexes associated with endothelial cell membranes, the coumarin, brodifacoum was given three days before in vitro experiments were carried out in order to inhibit production of active vitamin K1-dependent clotting factors. Brodifacoum (10 mg/kg i.p.) increased prothrombin time from 8.5 +/- 0.24 sec (control), to 46.2 +/- 2.4 sec (p less than 0.05), but had no effect on thrombin-induced relaxation (Emax greater than 90%; ED50 0.026 +/- 0.004 units/ml control; 0.025 +/- 0.004 unit/ml brodifacoum). In the final group of studies, we investigated the effects of the concomitant administration of protein C (1000 ng/ml) and thrombin in vitro. Protein C (1000 ng/ml) increased relaxant sensitivity to thrombin after partial desensitization of the relaxant response by previous thrombin administration, (-60.2 +/- 7.1% thrombin alone; -77.9 +/- 7.2% thrombin + protein C), but had no effect after complete desensitization of the relaxant response. In conclusion, the data appear best explained by protein C and thrombin-induced EDRF dependent relaxation being due to proteolytic actions.

4-Hydroxycoumarins↗