[Physiological phenomena and excluded volume effect--macromolecule activity coefficient].
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Animals↗
Biomedical subjects
Yoshiteru Seo
Publications and source records attributed to Yoshiteru Seo.
Dual effects of n-alcohols on fluid secretion from guinea pig pancreatic ducts.
Ethanol strongly augments secretin-stimulated, but not acetylcholine (ACh)-stimulated, fluid secretion from pancreatic duct cells. To understand its mechanism of action, we examined the effect of short-chain n-alcohols on fluid secretion and intracellular Ca(2+) concentration ([Ca(2+)](i)) in guinea pig pancreatic ducts. Fluid secretion was measured by monitoring the luminal volume of isolated interlobular ducts. [Ca(2+)](i) was estimated using fura-2 microfluorometry. Methanol and ethanol at 0.3-10 mM concentrations significantly augmented fluid secretion and induced a transient elevation of [Ca(2+)](i) in secretin- or dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP)-stimulated ducts. However, they failed to affect fluid secretion and [Ca(2+)](i) in unstimulated and ACh-stimulated ducts. In contrast, propanol and butanol at 0.3-10 mM concentrations significantly reduced fluid secretion and decreased [Ca(2+)](i) in unstimulated ducts and in ducts stimulated with secretin, DBcAMP, or ACh. Both stimulatory and inhibitory effects of n-alcohols completely disappeared after their removal from the perfusate. Propanol and butanol inhibited the plateau phase, but not the initial peak, of [Ca(2+)](i) response to ACh as well as the [Ca(2+)](i) elevation induced by thapsigargin, suggesting that they inhibit Ca(2+) influx. Removal of extracellular Ca(2+) reduced [Ca(2+)](i) in duct cells and completely abolished secretin-stimulated fluid secretion. In conclusion, there is a distinct cutoff point between ethanol (C2) and propanol (C3) in their effects on fluid secretion and [Ca(2+)](i) in duct cells. Short-chain n-alcohols appear to affect pancreatic ductal fluid secretion by activating or inhibiting the plasma membrane Ca(2+) channel.
1-Propanol↗
Sequence of forebrain activation induced by intraventricular injection of hypertonic NaCl detected by Mn2+ contrasted T1-weighted MRI.
In order to define the sequence of forebrain activation involved in osmoregulation, central activation in response to intracerebroventricular injection of NaCl solution (10 microl of 0.15, 0.5, or 1.5 M) was detected using manganese-contrasted magnetic resonance imaging (MRI) in anesthetized rats. Changes in renal sympathetic nerve activity (RNA) were also measured, and the time courses of forebrain activation and RNA changes compared. NaCl injection resulted in rapid activation of the subfornical organ (SFO), organum vasculosum lamina terminalis (OVLT), and periventricular regions and the lateral hypothalamic area (LHA), then of the paraventricular hypothalamic nucleus (PVN) and supraoptic nucleus (SON). The delay in activation in the PVN and SON showed a wide variation from 0 to 5.78 min, and the average delay in the PVN (2.88+/-0.34 min) and SON (2.90+/-0.39 min) was significantly greater than that in the SFO (0.40+/-0.10 min) and OVLT (0.74+/-0.13 min). NaCl (1.5 M) injection elicited a rapid, large increase in RNA, which consisted of two components, an early rapid increase at 99 s after injection (160+/-27%) and a slower increase at 9 min after injection (209+/-34%). These results suggest that the PVN and SON are activated not only by the afferent input from the SFO and OVLT but also by diffusion of the hypertonic stimulus to these regions and probably by their intrinsic osmosensitivity. The PVN might be responsible for the second slower component of the RNA response, but cannot be responsible for the first component.
Animals↗
Size-selective loosening of the blood-brain barrier in claudin-5-deficient mice.
Tight junctions are well-developed between adjacent endothelial cells of blood vessels in the central nervous system, and play a central role in establishing the blood-brain barrier (BBB). Claudin-5 is a major cell adhesion molecule of tight junctions in brain endothelial cells. To examine its possible involvement in the BBB, claudin-5-deficient mice were generated. In the brains of these mice, the development and morphology of blood vessels were not altered, showing no bleeding or edema. However, tracer experiments and magnetic resonance imaging revealed that in these mice, the BBB against small molecules (<800 D), but not larger molecules, was selectively affected. This unexpected finding (i.e., the size-selective loosening of the BBB) not only provides new insight into the basic molecular physiology of BBB but also opens a new way to deliver potential drugs across the BBB into the central nervous system.
Animals↗
Evaluation of collagen fiber maturation and ordering in regenerating tendons employing H-1 double quantum filtered NMR spectroscopy.
It is difficult to monitor the chronic stage of the healing process of ruptured tendons employing the present diagnostic modes. However, the results of this study have shown that 1H double quantum filtered (DQF) NMR spectroscopy is sensitive to the later stages of the healing process. Regenerated tendons of rabbits were dissected and measured at the end of the acute phase (three weeks), the subacute phase (nine weeks), and the chronic phase (13 and 18 weeks after tenotomy). Four parameters were determined by 1H DQF NMR spectroscopy: (a) the maximum signal intensity (h(max)) relative to the single quantum spectrum, (b) the creation time of the maximum signal intensity (tau(max)), (c) the decay time from the maximum signal intensity to a value half of that intensity (tau(1/2)) and (d) the residual dipolar splitting of water (delta), representing the order of the collagen fibers. The values of h(max), tau(max), tau(1/2), and delta of the intact Achilles tendons were 11.3+/-1.0%, 0.48+/-0.03 ms, 0.67+/-0.04 ms and 732+/-62 Hz (mean +/- SEM, n=6), respectively. In the regenerating tendon, h(max) increased from 0.41+/-0.12% at three weeks to 7.07+/-0.77% at 18 weeks, tau(max) decreased from 1.88+/-0.31 ms at three weeks to 0.72+/-0.04 ms at 18 weeks, tau(1/2) decreased from 11.6+/-1.8 ms at 3 weeks to 1.48+/-0.16 ms at 18 weeks, and delta increased from 129+/-8 Hz at three weeks to 414+/-29 Hz at 18 weeks. We have concluded that reordering of collagen fibers proceeds continuously even in the chronic stage of healing. Thus, the 1H DQF NMR spectroscopy is a useful non-invasive technique to evaluate the reconstruction and the order of collagen fibers in regenerating tendon. It is also suggested that tau(1/2) and h(max) are most useful for in vivo DQF NMR spectroscopy and imaging, respectively, in combination with tau(max).
Achilles Tendon↗
Water permeability of capillaries in the subfornical organ of rats determined by Gd-DTPA(2-) enhanced 1H magnetic resonance imaging.
The water permeability of capillaries in the subfornical organ (SFO) of rat was measured by a (1)H nuclear magnetic resonance method in combination with a venous injection of a relaxation reagent, gadolinium-diethylene triamine-N,N,N',N",N"-pentaacetic acid (Gd-DTPA(2-)), which could not pass through the blood-brain barrier (BBB). Judging from results of Gd-DTPA(2-) dose dependency in the intact brain and the BBB-permeabilized brain, Gd-DTPA(2-) could not have leaked out from the capillaries in the cortex, thalamus or SFO, but it could have been extravasated in the posterior lobe of the pituitary gland. The longitudinal (T(1)) relaxation time of water in the SFO region was measured by inversion-recovery magnetic resonance imaging at 4.7 T. The T(1) relaxation rates (1/T(1)) before and after Gd-DTPA(2-) infusion were 0.70 +/- 0.02 s(-1) (mean +/- S.E.M., n = 9) and 1.53 +/- 0.11 s(-1) (n = 9), respectively. The rate constant for water influx to the capillaries was estimated to be 0.84 +/- 0.11 s(-1) (n = 9) which corresponds with a diffusive membrane permeability (P(d)) of 3.7 x 10(-3) cm s(-1). Compared with values found in the literature available on this subject, this P(d) value for the capillaries in the SFO was the same order of magnitude as that for transmembrane permeability of water for the vasa recta, and it may be 10-100 times larger than that of the blood-brain barrier in the cortex. Areas of the cortex and thalamus showed minimal changes in the T(1) relaxation rate (ca 0.09 s(-1)), but these values were not statistically significant and they corresponded to P(d) values much smaller than those found in the SFO. From these results, we conclude that the capillaries in the SFO have one of the highest water permeability values among all of the capillaries in the brain. It is also suggested that this magnetic resonance imaging, based on T(1) relaxation rate, is a useful method to detect local water permeability in situ.
Animals↗
Detection of hypothalamic activation by manganese ion contrasted T(1)-weighted magnetic resonance imaging in rats.
To examine the usefulness of Mn(2+) contrasted magnetic resonance imaging (MRI) in hypothalamic imaging, images obtained using T(1)-weighted MRI were compared with Fos expression, which is known to increase after activation of voltage-dependent Ca(2+) channels. Intravenous infusion of MnCl(2) elicited a rapid increase in the T(1)-weighted MRI signal intensity in the vessels and ventricles, but not in the brain parenchyma, suggesting that Mn(2+) did not diffuse freely across the blood-brain barrier. When the blood-brain barrier was broken by right intracarotid arterial injection of 25% D-mannitol, an increased signal intensity was seen in the right brain. Intracarotid arterial injection of hypertonic NaCl elicited rapid and striking increases in signal intensity in the paraventricular hypothalamic nucleus, supraoptic nucleus, and preoptic area, which are thought to be involved in central osmotic regulation. These observations were consistent with the Fos expression results. These results indicate that Mn(2+) contrasted MRI is a useful technique for investigating the autonomic center in the hypothalamus.
Animals↗
Mapping the fiber orientation in articular cartilage at rest and under pressure studied by 2H double quantum filtered MRI.
The one-dimensional (2)H double quantum filtered (DQF) spectroscopic imaging technique was used to study the orientation of collagen fibers in articular cartilage. The method detects only water molecules in anisotropic environments, which in cartilage is caused by their interaction with the collagen fibers. A large quadrupolar splitting was observed in the calcified zone and a smaller splitting in the radial zone. In the transitional zone the splitting was not resolved and a small splitting was again detected in the superficial zone. From measurements performed at two orientations of the plug relative to the magnetic field it was deduced that in the calcified and radial zones the fibers are oriented perpendicular to the bone, bending at the transitional zone and flattening at the superficial zone. The effect of load applied to the cartilage-bone plug was monitored by the same technique. At low loads there is a small decrease in the quadrupolar splitting in the calcified zone, a marked decrease in the radial zone, and an increase of the splitting accompanied by a thickening of the superficial zone. Under high loads, while the thickening and the splitting of the superficial zone further increase, the splitting in the radial and calcified zones completely collapse. Pressure-induced changes in the thickness of the surface zone indicate flattening of the collagen fibers near the surface. The marked collapse of the splitting near the bone at high pressures may result from crimping of the collagen fibers.
Animals↗