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Biomedical subjects

I Sakuma

Publications and source records attributed to I Sakuma.

At least 91 records · Page 5Linked to original sources

High-performance liquid chromatographic determination of catecholamine metabolites and 5-hydroxyindoleacetic acid in human urine using a mixed-mode column and an eight-channel electrode electrochemical detector.

An HPLC system for the simultaneous determination of acidic catecholamine metabolites, related compounds and 5-hydroxyindoleacetic acid (5-HIAA) in human urine was developed. A mixed-mode (C18/anion-exchange) column with isocratic elution using citrate buffer and an eight-channel electrochemical detector were used. Vanilmandelic acid (VMA), 3,4-dihydroxyphenylacetic acid (DOPAC), 4-hydroxy-3-methoxyphenyllactic acid (vanillactic acid, VLA), homovanillic acid (HVA), vanillic acid (VA) and 5-HIAA in urine were determined simultaneously. Detection limits and inter (n = 5) and intra-assay (n = 5) coefficients of variation were satisfactory. The mean of analytical recoveries (n = 3, +/- C.V. (%)) were between 97 +/- 3.2 (VMA) and 105 +/- 4.8 (VA). Correlations between the analytical results for VMA, HVA and 5-HIAA obtained by an established method and the present method were satisfactory. The mean +/- 2 S.D. of the excretion rates of VMA, DOPAC, VLA, HVA, 5-HIAA and VA in urine from healthy adult volunteers were 0.61-4.36, 0.13-1.02, 0-0.35, 0.67-6.55, 0.50-5.14 and 0-0.55 mg/g creatinine, respectively.

3,4-Dihydroxyphenylacetic Acid↗

Laminar flow stimulates ATP- and shear stress-dependent nitric oxide production in cultured bovine endothelial cells.

Based on the fact that nitric oxide (NO) production is associated with changes in intracellular cGMP levels and is selectively inhibited by N omega-methyl L-arginine (L-NME), we investigated the shear stress dependency of NO production in endothelial cells (ECs) from its cGMP responses to various shear stress loads. Cultured fetal bovine aortic ECs treated with a phosphodiesterase inhibitor, isobutylmethylxanthine (IBMX; 1 mM), were exposed to a laminar flow of Krebs buffer solution for 5 minutes in a parallel-plate flow chamber and examined for changes in intracellular cGMP levels by radioimmunoassay using an [125I] cGMP kit. Application of flow increased the cGMP levels. The increase was significant in the presence of extracellular ATP (1 microM)(control, 286.1 +/- 43.6; flow, 506.5 +/- 44.9 fmol/10(7) cells; p < 0.001), but not in its absence (control, 256.6 +/- 60.6; flow, 301.5 +/- 91.4 fmol/10(7) cells; N.S.). The cGMP levels increased significantly as the magnitude of shear stress applied increased. Treatment of ECs with a specific inhibitor of NO production, L-NMA (200 microM), completely inhibited the flow-induced increase in cGMP, and L-arginine reversed the L-NMA-induced inhibition, indicating that the increase in cGMP was due to NO produced by the flow. The flow-induced increase in NO production was markedly suppressed when extracellular Ca++ was chelated by adding EGTA to the perfusate. These findings suggest that flow stimulates NO production to increase cGMP levels shear stress-dependently in ECs and that extracellular Ca++ and ATP modulate the effects of flow.

1-Methyl-3-isobutylxanthine↗

Depressed responsiveness to angiotensin II in ventricular myocytes of hypertrophic cardiomyopathic Syrian hamster.

Electromechanical responsiveness to angiotensin II (Ang II) receptor stimulation in ventricular myocardium and myocytes of hypertrophic cardiomyopathic Syrian hamsters (BIO 14.6) was examined and compared with that in preparations of normal hamsters (F1B) using conventional microelectrode and patch clamp techniques. Action potential duration (APD) and developed tension (DT) corrected for the cross-sectional area of the papillary muscles of 14-20 week-old BIO 14.6 hamsters were significantly smaller than those in preparations of age-matched normal hamsters. An Ang II (1 microM)-induced increase in DT in BIO 14.6 papillary muscles (24.7 +/- 11.0%) was significantly smaller than that in F1B papillary muscles (53.8 +/- 8.5%), which was associated with a smaller increase in APD in BIO 14.6 papillary muscles. In ventricular myocytes of both BIO 14.6 and F1B hamsters. Ang II increased the calcium current (ICa) following a transient decrease in ICa. However, the magnitude of the Ang II-induced increase in ICa in BIO 14.6 myocytes (35.5 +/- 7.5%) was significantly smaller than that in F1B myocytes (86.0 +/- 19.7%), suggesting a causal relationship between ICa and mechanical response to Ang II in these hamsters. The depressed responsiveness to Ang II receptor stimulation in hypertrophic cardiomyopathic hamster is in a marked contrast with the enhanced responsiveness to alpha 1-adrenergic stimulation, which was demonstrated by previous studies, and may be one of adaptational changes to the activated renin-angiotensin system in the cardiomyopathy.

Action Potentials↗

Notes on the acetylcholine-induced relaxation of porcine coronary arteries.

Intact and rubbed coronary arteries responded to acetylcholine (ACh) with contractions in a cumulative dose-dependent manner. The rubbed artery contracted at a lower concentration of ACh (3 x 10(-8) M) than the threshold level of the agonist in the intact artery (10(-7) M). Only in the intact artery was the second cumulative dose-response to ACh (10(-6) M) decreased by 60% (p < 0.05). Pretreatment of the intact artery with methylene blue or NG-monomethyl-L-arginine (L-NMMA) increased the contractile response to ACh. The ACh-induced increase of the cGMP level in the intact artery was eliminated by removal of the endothelial cells and by pretreatment of the artery with either methylene blue or L-NMMA. These findings indicate that the endothelial cells in the coronary artery are responsible for the ACh-induced relaxation, presumably mediated by an endothelium-derived relaxing factor.

Acetylcholine↗

Hemispheric representation of the central retina of commissurotomized subjects.

It is controversial whether a stimulus projected within 1 to 3 degrees from the boundary between the right and left hemiretina is transmitted to only one cerebral hemisphere or to both cerebral hemispheres. In order to resolve this issue, letter- and word-stimuli were presented for 200 msec with a new type of tachistoscope, called the fundus tachistoscope, in and about the central retina, (i.e. fovea, 1.2 degrees in horizontal diameter) of the right eyes of two commissurotomized subjects (N.G. and A.A.). During stimulus presentation the subjects were attempting to fixate a fixation target. The fundus tachistoscope combined with image analysis of the fundus enables us to measure the position of the stimulus on the retina, relative to the foveal center, as well as whether or not the eye moved during stimulus presentation. The results indicate that the region of the right (temporal) hemiretina represented by both hemispheres in letter processing, if it exists, was estimated as less than 0.6 degrees from the foveal center. The two subjects frequently (27% in N.G. and 46% in A.A.) fixated the fixation target eccentrically, i.e. with a retinal point other than the foveal center, during fixation, namely stimulus presentation. Their eccentric fixations were small with magnitude almost all falling between 1.35 degrees right and 1.25 degrees left of the foveal center. It is therefore recommended that letter-stimuli be presented at least 2.0 degrees from the foveal center in ordinary tachistoscopic studies of cerebral hemispheric differences. Eye movements, which varied in 0.11 degrees and 1.43 degrees horizontally, occurred in about 8% of all the trials during fixation. On the average of the two subjects, the eye movements caused or worsened eccentric fixation in only about one third of the trials, and corrected eccentric fixation in about two thirds of the trials.

Adult↗

Lysophosphatidylcholine, a component of stromal phospholipids, as a candidate vasoconstrictive factor in stroma-free hemoglobin.

Stroma-free hemoglobin (SFH) contains a trace amount of phospholipids, which has been implicated in the toxic reactions associated with SFH. We analyzed stromal phospholipids by high-performance liquid chromatography and found that SFH contained small quantities of lysophosphatidylcholine (LPC), which is known to be capable of producing a defect in endothelium-dependent arterial relaxation, in addition to major classes of constituent phospholipids in red cell membrane. LPC content was determined to be 1.65 nmol/ml (hemoglobin 8.1 g/dl). To evaluate the role of these stromal phospholipids in SFH-induced vasoconstriction, we next examined the effect of lipids on vascular tone in rabbit aortic strips. Preincubation with the crude lipid extract or the LPC purified from SFH significantly inhibited acetylcholine-induced relaxation in phenylephrine-precontracted tissues. The LPC-induced inhibition was reversed by incubation of the tissues in the absence of lipids, indicating the functional integrity of endothelium. From these results, we propose a possibility that LPC, a component of stromal phospholipids, is a candidate for vasoconstrictive factors present in SFH.

Acetylcholine↗

Internal hydraulic loss in a seal-less centrifugal Gyro pump.

A new index "loss factor Z" defined by Eq. 1 was introduced as the absolute expression of the mock loop resistance for testing a nonpulsatile pump. [formula: see text] where gamma is specific gravity of the fluid, g is the acceleration of gravity, delta P is total pressure head, and Q is flow. Z is expected to be constant, regardless of the pumping parameters. Z values obtained in the same mock loop but with different rotary blood pumps were almost identical and were defined as Z0. New methods of analysis of the flow-restrictive conditions of various rotary blood pumps are proposed in this paper: namely, differential loss factor delta Z, and loss factor sensitivity delta Z/delta A. The proposed Z-Q curves demonstrated better performance mapping than the conventional delta P-Q curves. Delta Z is the difference between the Z-Q curves of two different pumps. A is a design parameter of the pump; therefore delta Z/delta A is a quantitative expression of the effect of the design change on the hydraulic performance. These various indices were used to analyze the internal hydraulic loss of a centrifugal pump (Gyro pump). The relationship between its gap size (rotor casing) and hydraulic performance was assessed quantitatively by these indices. In this paper, the derivation processes and above-mentioned indices are described.

Assisted Circulation↗

Aftereffects of high-intensity DC stimulation on the electromechanical performance of ventricular muscle.

To clarify the mechanisms underlying cardiac dysfunction after electrical defibrillation, we investigated the effects of direct current field stimulation (10 ms, 1-80 V/cm) on isolated guinea pig papillary muscles. Shocks (S2) > 15 V/cm lowered the plateau height of the S2-induced action potential and inhibited its terminal repolarization. Subsequent responses to basic stimuli (S1, 1.0 Hz) for 1-3 min were characterized by a decrease in the maximum diastolic potential, a shortening of action potential duration, and an increase of the developed tension. With S2 > 30 V/cm, a marked delay in repolarization of the S2-induced action potential was followed by oscillation of membrane potential, resulting in repetitive spontaneous activity and often refractoriness to S1 stimulation. The aftereffects were independent of the phase of S2 application. Most of the aftereffects were preserved in the presence of nifedipine (1 microM) or ryanodine (1 microM). Only sodium channel blockade by tetrodotoxin (10 microM) modified the aftereffects by depressing the generation of spontaneous activity. These findings suggest that strong shocks (> 15 V/cm) will produce abnormal arrhythmogenic responses probably through a transient rupture of sarcolemmal membrane (electroporation) leading to a disturbance of the ionic equilibrium of the myocyte.

Animals↗

The quality control of stroma-free hemoglobin: lysophosphatidylcholine, a component of stromal phospholipids, as candidate vasoconstrictive factor.

We characterized stromal phospholipids in stroma-free hemoglobin (SFH) by normal-phase and cation-exchange HPLCs, and found that SFH contained not only four phospholipids which were the major constituent classes in membrane, but also several peaks which were not yet identified. The residual amounts of these lipids in SFH were changed with storage of red cell concentrates. The four major phospholipids decreased concomitantly with storage, whereas the unidentified peaks increased after 21 days and then decreased after 48 days. We also found that SFH contained lysophosphatidylcholine (LPC) at 5.71 micrograms/ml, which was the deacylated metabolite of phosphatidylcholine (PC). These results suggest that stromal phospholipids are degradable. Since LPC is known to be capable of producing a defect in endothelium-dependent arterial relaxation, we next examined the effect of stromal lipids on vascular tone in rabbit thoracic aortic strips. Preincubation with the crude lipid extract or the LPC purified from SFH by TLC significantly inhibited acetylcholine (ACh)-induced relaxation in phenylephrine (PhE)-precontracted tissues. These observations have led to the proposal that LPC, a component of stromal phospholipids, induces vasoconstriction as a result of inhibition of endothelium-dependent vasorelaxation.

Animals↗

[Fluctuation of HCV quasi-species population during interferon therapy; analysis by single strand conformation polymorphism].

We investigated the fluctuation of hepatitis C virus (HCV) quasispecies during interferon therapy by single strand conformation polymorphism (SSCP) analysis. In 13 of 16 interferon ineffective patients, the predominant HCV population was replaced with other quasispecies during the treatment. Especially, in 9 patients, a part of the HCV quasispecies, pre-existing before interferon therapy, became predominant after the therapy. These results indicate that sensitivity to interferon differs among HCV quasispecies and that interferon selects resistant HCV strains. Existence of such HCV quasispecies seems to be associated with interferon treatment failure.

Base Sequence↗

[Image diagnosis of vasculitis--ultrasound].

Ultrasound is widely used for the diagnosis and follow-up of patients with many kinds of vasculitis. Transcutaneous or transesophageal two-dimensional ultrasound images can depict vascular wall lesions associated with vasculitis such as stenosis, aneurysm or wall thickening. Pulsed Doppler and color Doppler are useful for the assessment of stenoses and changes in blood flow. Recent progress in intravascular ultrasound apparatus made it possible to more precisely assess morphological changes in vascular wall. Intravascular ultrasound of a 27-year old male patient suffering from inferior myocardial infarction with right coronary aneurysms revealed calcification and fibrosis of the intima and early development of atherosclerosis in both right and left coronary arteries suggestive of the sequelae of Kawasaki disease.

Adult↗

Pertussis toxin pretreatment alters agonist binding to beta-adrenoceptors in rabbit ventricular myocardium.

The influence of pretreatment with pertussis toxin (PTX) on characteristics of beta-adrenoceptor bindings was examined in myocardial ventricular membranes prepared from rabbits which received an intravenous injection of PTX (10 micrograms/kg) or its vehicle. The density and affinity of beta-adrenoceptors assessed by (-)-[125I]iodocyanopindolol were not significantly altered by PTX pretreatment. However, a fraction of agonist binding sites with high affinity significantly increased in PTX-pretreated myocardial membranes. In the presence of 100 microM 5'-guanylyl imidodiphosphate, all agonist binding sites were converted to low-affinity sites in both control and PTX-pretreated membranes. The results suggest that Gi might regulate negatively coupling of beta-adrenoceptors to Gs.

Animals↗

The problem of macular sparing after unilateral occipital lesions.

Whether or not unilateral occipital damage produces sparing of central vision, namely macular sparing, is controversial. We tested two subjects with left occipital lesions by means of fundus perimetry combined with fundus image analysis. This method made it possible to measure the distance of the stimulus projected on the retina from the foveal centre defined as the centre of the foveal reflex. The results indicated that macular sparing, if it exists, must be less than 0.4 degree wide. Two of the four eyes during the stimulus presentation often but not always showed eccentric fixation of a small magnitude, whose mean was less than 0.6 degree from the foveal centre in the right hemiretina.

Cerebral Infarction↗

Possible contribution of potassium channels to the endothelin-induced dilatation of rat coronary vascular beds.

The mechanism for the endothelin (ET)-induced vasodilatation of the endothelium-intact rat coronary vascular bed was investigated. Continuous infusion (0.1-1 nM) or bolus injection (1-100 pmol) of ET-1 or ET-3 elicited a dose-related transient decrease, followed by a slight sustained increase, in the coronary perfusion pressure (CPP). The decrease in CPP induced by an injection (10 pmol) of ET-1 or ET-3 was not modified by indomethacin (5 microM). However, oxyhemoglobin (5 mM) shortened the duration of the ET-induced decrease in CPP, although it did not affect the magnitude. The ET-induced decrease in CPP was abolished by raising K+ in the perfusing solution from 5.9 to 16.9 mM. These findings suggest that the ET-induced dilatation of the rat coronary vascular beds may not be mediated by cyclooxygenase products. The ET-induced vasorelaxation may be mediated, at least in part, by endothelium-derived relaxing factor and may be related to opening of K+ channels.

Animals↗