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Biomedical subjects

Y Sakamoto

Publications and source records attributed to Y Sakamoto.

At least 271 records · Page 15Linked to original sources

Local delivery of antithrombotic drug prevents restenosis after balloon angioplasty in atherosclerotic rabbit artery.

We investigated the ability of various antithrombotic drugs, delivered locally, to prevent restenosis after angioplasty in hypercholesterolemic rabbits. After dilating atherosclerotic iliac stenoses by balloon angioplasty, a low dose of heparin or a new antithrombotic drug, such as low molecular weight heparin (fragmin), argatroban, or batroxobin, was delivered locally using the balloon double-occlusion technique. In 1 group, high-dose heparin was administered intravenously. Animals that received no drugs served as a control group. After angioplasty, the stenotic segment was dilated and the mean percentage luminal stenosis fell from 89% to 9% in the group that received locally delivered heparin, from 88% to 7% in the group that received locally delivered argatroban, from 87% to 11% in the group that received locally delivered fragmin, from 88% to 15% in the group that received locally delivered batroxobin, from 82% to 18% in the group that received i.v. heparin (p < 0.0001 compared with before angioplasty in each case), and from 84% to 17% in the control group (p < 0.005 compared with before angioplasty). Twenty-eight days after angioplasty, the percentage luminal stenosis remained at 14% in the group that received locally delivered argatroban, 15% in the group that received locally delivered fragmin, and 28% in the group that received locally delivered batroxobin, whereas it increased to 45% in the group that received i.v. heparin, 30% in the group that received locally delivered heparin and 72% in the control group (p < 0.05 compared with after angioplasty in each case). Thus, local delivery low doses of new antithrombotic drugs prevents restenosis after angioplasty without affecting systemic coagulability; heparin, whether administered locally or intravenously, was less effective than the new drugs in preventing restenosis.

Angiography↗

Effects of LHRH, FSH and activin A on follistatin secretion from cultured rat anterior pituitary cells.

We demonstrated the release of follistatin, an activin-binding protein, from cultured rat anterior pituitary cells by measuring immunoreactive (ir-) follistatin in a specific immunoradiometric assay. Ir-follistatin release gradually increased in cultures over 1-18 days and reached its maximal level at 12-15 days of incubation. The basal ir-follistatin levels in the culture media increased about 3- (P < 0.01) and 5-fold (P < 0.001) in 2 and 10% fetal calf serum for 6 days, respectively. LHRH and activin A caused an approximately 2.0- (P < 0.05) and 1.8-fold (P < 0.05) rise in ir-follistatin release, respectively, in contrast to the lack of significant FSH effects. The culture medium condensed on sulfate-cellulofine gel was resolved by polyacrylamide gel electrophoresis and blotted with anti-follistatin polyclonal antibody, resulting in at least three protein bands ranging from 35 to 50 kDa under non-reducing conditions. These results indicated that follistatin is produced in anterior pituitary cells and that its secretion is regulated at least in part by LHRH and activin, implying an autocrine/paracrine role of activin and follistatin in the pituitary.

Activins↗

Regulation of immunoreactive activin A secretion from cultured rat anterior pituitary cells.

Immunoreactive activin A (ir-activin A) release from cultured rat anterior pituitary cells was examined by measuring ir-activin A in culture medium by a specific radioimmunoassay. Ir-activin A release into the medium increased over 1-18 days, and reached a maximal level at 12-15 days. The basal levels of ir-activin A in the culture media were 0.70 +/- 0.10 (mean +/- SD), 1.30 +/- 0.36 and 1.83 +/- 0.44 ng/10(6) cells, when cultured for 6 days with 0, 2 and 10% fetal calf serum, respectively. LHRH induced an approximate 1.4-fold increase in ir-activin A release in contrast to a 40-60% inhibition with FSH, but LH did not affect the activin A release. In the presence of 12-o-tetradecanoylphorbol acetate (TPA), iractivin A release was enhanced, but no significant effect was induced by forskolin. Activin A was distinctly immunostained in cultured rat anterior pituitary cells. These results suggested that activin A release from the pituitary is modified by FSH and LHRH, and that the activation of protein kinase C may be involved in the action of LHRH.

Activins↗

Determination of free follistatin levels in sera of normal subjects and patients with various diseases.

We developed an assay system for measuring free follistatin by using an anti-follistatin mouse monoclonal antibody and [125I]activin A. The sensitivity of this assay was 0.5 microgram/l and cross-reactivities with inhibin, luteinizing hormone, follicle-stimulating hormone and growth hormone were all less than 0.5%. The dose-response curves of human sera and follicular fluid were parallel to the standard curve, and the follicular fluid contained a large amount of follistatin (6.4 +/- 0.5 mg/l, mean +/- SEM; N = 13). The within- and between-assay coefficients of variation calculated from the analysis of serum samples of four different concentrations were 3.3-7.8% and 3.9-11.0%, respectively. The recovery rates of free follistatin at five different doses were 86.4 - 102.4%. When activin A was added to the same sample, free follistatin recovery rate declined dose-dependently. Gel filtration analyses of human serum and follicular fluid resulted in a single peak corresponding to authentic follistatin. Using this assay, free follistatin concentrations in sera were measured in normal, pregnant and diseased subjects. The free follistatin level in serum of normal adults was 3.5 +/- 0.2 micrograms/l (N = 60), which was significantly elevated in pregnant women (16.7 +/- 1.3 micrograms/l, N = 56), and in patients with chronic liver disease (8.1 +/- 1.1 micrograms/l, N = 20), chronic renal failure (6.7 +/- 0.9 micrograms/l, N = 42), advanced solid cancer (8.5 +/- 1.0 micrograms/l, N = 39) and hematological malignancies (6.8 +/- 1.0 micrograms/l, N = 18). These data indicated that the free follistatin concentration in serum is detectable and varies during pregnancy and in various diseased states.

Adult↗

Inhibition of chemiluminescence response of human polymorphonuclear leukocytes by three different stimulations in hyperosmotic condition comparable to the renal medulla.

Hyperosmolality in renal medulla inhibits functions of polymorphonuclear leukocyte (PMN) such as phagocytosis, intracellular killing, and superoxide generation. The main factors of hyperosmolality in the renal medulla are thought to be urea and NaCl. We studied the luminol-dependent chemiluminescence (CL) response of PMNs to three different stimulators: phorbol myristate acetate (PMA), formyl-methionyl-leucyl-phenylalanine (FMLP), and opsonized zymosan (OZ). When PMNs were incubated or preincubated with hyperosmotic urea or NaCl solutions, CL responses were significantly reduced following stimulation by each of PMA, FMLP, or OZ. Reduction of CL response was concentration and osmolality dependent in hyperosmotic urea and NaCl solution. These results suggest that respiratory burst and production of active oxygen species of PMNs through three different signal transduction pathways are inhibited in hyperosomotic conditions comparable to the renal medulla.

Cells, Cultured↗

Visual acuity disturbance in subjects over 50 years of age in a population-based cataract survey.

Visual acuity is still an essential examination item in cataract epidemiological studies, even though this parameter lacks objective reliability. A population-based epidemiological study was conducted in a rural area of Japan to find out the relationship between visual acuity levels and cataracts, the types of gradings of which were evaluated by an objectively reliable method through lens images, and to serve as a sample for researchers who perform epidemiological studies on cataract but lack the latest methodology. 863 participants above 50 years of age were examined and those previously diagnosed with ocular diseases which affect visual acuity were excluded from the analysis as much as possible. The mean visual acuity (LogMAR) in subjects in their 50s, 60s, 70s and over 80 years of age was 1.1, 1.0, 0.7 and 0.4, respectively. Both the mean and the distribution of visual acuity showed a statistically significant relationship to ageing (p < 0.01, p < 0.01). Both age and cataract grading showed a statistically significant relationship to visual acuity (p < 0.01), but there was no significant interaction effect between these two variables upon visual acuity. Visual acuity worsened remarkably with ageing in the eyes with grade III, whereas that of eyes with grades I and II remained fairly stable at a level of 0.7-1.0, except for those above 80 years of age. A fairly good visual acuity of 1.0, 0.8 and 0.7 remained in the eyes with pure cortical cataract of grades I, II and III, respectively, whereas those with mixed type cataract were 0.9, 0.6 and 0.3, respectively.

Aged↗

Assessment of mitochondrial gene in proliferative vitreoretinal tissues from patients with familial diabetes mellitus.

Fibrovascular tissues obtained at therapeutic vitrectomy from 22 patients with proliferative diabetic retinopathy were studied for a mutation of tRNALEU(UUR) at nucleotide position (np) 3243 of mitochondrial DNA (mtDNA). We found a mutation of mtDNA in the vitreous sample of one patient, a 44-year-old woman who had maternally inherited, familial, non-insulin-dependent diabetes mellitus of 25-year duration with recurrent vitreous hemorrhages due to proliferative retinopathy. Heteroduplex analysis also detected the mutant-type mtDNA in the vitreous sample of this patient. The abnormality was, however, not observed in the peripheral blood sample from either this patient orr her family members with diabetes mellitus. The other 21 patients were negative for the mutation in vitreous specimens as well as in peripheral blood. Although a firm conclusion cannot be drawn from a single case report, our observations suggest that this case was an example of tissue-specific expression of mtDNA mutation. Further studies of a larger number of patients with familial diabetes mellitus seem justified.

Adult↗

[Saccular aneurysm associated with fenestration of the distal segment of basilar artery].

A 32-year-old man who presented with subarachnoid hemorrhage was found to have a saccular aneurysm arising from the proximal end of the fenestration formed at the distal third of basilar artery. The aneurysm dome was directed anteriorly and measured 3mm in diameter. It was successfully clipped via subtemporal transtentorial approach and the patient returned to his previous job. Basilar artery fenestration is angiographically found in 0.23-0.6%. The fenestration is most frequently located in the proximal basilar trunk close to the vertebro-basilar junction. In this report a rare case with an aneurysm associated with fenestration in the distal part of the basilar artery will be presented.

Adult↗

Taurine in the liver. The function of taurine conjugated with bile acids.

1. Taurine had cytoprotective effects on hepatocytes under hyperoxic or hypoxic condition in the presence of Ca2+. 2. Bile acids had toxic effects on hepatocytes; there was no significant difference in the toxicity between free bile acids and taurine-conjugated bile acids. 3. The mobility of the membranes, examined by 31P-NMR, decreased after treatment with taurine or free bile acids, but increased after treatment with taurine-conjugated bile acids. 4. The fluidity of the membranes, measured by ESR, did not change after treatment with taurine, but decreased after treatment with free bile acids and increased after treatment with taurine-conjugated bile acids. In conclusion, because most bile acids are conjugated with taurine in the liver after administration of taurine, the influence of taurine-conjugated bile acids has to be taken into consideration in estimating the functions of taurine in the liver.

Animals↗

[Clinical evaluation of urinary leukotriene E4 levels in asymptomatic bronchial asthma].

To evaluate the significance of peptide leukotrienes (LTC4, D4, E4) in asymptomatic asthmatic patients, we measured urinary LTE4 levels which is thought to reflect in vivo production of peptide LTs. Urinary LTE4, was extracted using C18 solid phase column and measured by radioimmunoassay. There was no significant difference in urinary LTE4 levels among asthmatics with different severity or between atopic and non-atopic asthmatics. Urinary LTE4 levels were significantly elevated in asthmatics compared with normal controls (p < 0.05). When compared with normal controls, urinary LTE4 levels were significantly elevated in moderate to severe asthma (p < 0.05), and non-atopic asthmatics (p < 0.001). Urinary LTE4 levels were significantly elevated in aspirin-sensitive asthmatics compared with aspirin-tolerant asthmatics (p < 0.05). There was no significant difference in urinary LTE4 levels among aspirin-sensitive asthmatics with different severity. These results suggest that increased production of peptide LTs is a characteristic in aspirin-sensitive asthma, and that the severity and type of asthma and the presence of aspirin-sensitive asthma should be taken into consideration in the analysis of urinary LTE4 levels.

Adult↗

Synergistic growth stimulation of mouse fibroblasts by tumor-derived adhesion factor with insulin-like growth factors and insulin.

Tumor-derived adhesion factor (TAF) has been purified as a major secretory protein with cell-adhesion activity from culture medium conditioned by human bladder carcinoma cells (K. Akaogi et al., Biochem. Biophys. Res. Commun., 198: 1046-1053, 1994). TAF is closely related or identical to a putative protein encoded by the mac25 gene (M. Murphy et al., Cell Growth & Differ., 4: 715-722, 1993) and prostacyclin-stimulating factor (T. Yamauchi et al., Biochem. J., 303: 591-598, 1994). TAF has a characteristic cysteine cluster in its NH2-terminal sequence, which is conserved in the NH2 termini of insulin-like growth factor-binding proteins. In the present study, we examined the interaction of TAF with insulin-like growth factors (IGFs) and insulin. Although in a ligand blotting assay TAF bound neither IGFs nor insulin, immunoprecipitation assay showed weak interaction of TAF with IGF-I, IGF-II, and insulin. TAF by itself stimulated the growth of BALB/c3T3 cells on the usual plastic substrates and significantly enhanced IGF- or insulin-mediated cell growth. Moreover, marked potentiation of the mitogenic activities of IGFs and insulin by TAF was found in the culture on type IV collagen substrates. The synergistic growth stimulation was completely blocked by heparin. These results suggest that TAF may regulate the growth of fibroblastic cells in collaboration with IGFs and insulin.

3T3 Cells↗

[FLAIR images of subarachnoid hemorrhage].

We studied MR fluid attenuated inversion recovery (FLAIR) pulse sequences in 37 cases with subarachnoid hemorrhage caused by aneurysmal rupture. FLAIR sequence suppressed the CSF signal and produced very heavy T2 weighted images. This has been of particular benefit in identifying lesions at the margins of the brain, around the basal cisterns, in the brain stem and at the gray-white matter junction. Based on these findings, we were able to detect the subarachnoid hemorrhage. Subarachnoid hemorrhage was able to be demonstrated as high signal intensity on FLAIR sequences in all patients. Clear visualization of acute subarachnoid hemorrhage was able to be obtained by MR FLAIR sequences in not only Fisher's group 3 or 4, but also Fisher's group 2. Moreover it was suited for the detection of intraaxial hematoma, Sylvian hematoma, subdural hematoma and subarachnoid hemorrhage in the posterior fossa and interhemispheric fissure. Especially, it was useful for detecting intraventricular hemorrhage. Therefore, if patients suffering from subarachnoid hemorrhage present slight headache or atypical symptoms, sometimes it may be more suitable to perform MRI FLAIR pulse sequences first. Aneurysms were found in 21 cases (56.8%). When the aneurysmal size is more than 7 mm, the rate of detection becomes 100%. Aneurysms present various MR appearances because of flow characteristics. Aneurysms were demonstrated as low signal intensity except in 3 cases. In one out of 3 cases, aneurysms were revealed as high signal intensity and in the other two cases, it was revealed as mixed signal intensity. According to the previous studies, rapid flow was demonstrated as low signal intensity by vascular flow void, and delayed flow was demonstrated as high or mixed signal intensity by flow related enhancement and even echo rephasing. MR clearly delineates the size, the lumen, the flow, and the extraaxial location of aneurysms. In conclusion, MR FLAIR pulse sequences could be reliable and useful for diagnosis of subarachnoid hemorrhage, especially as screening tests for outpatients.

Adult↗

Structural study of the glycosylated and unglycosylated forms of a genetic variant of human serum albumin (63 Asp-->Asn).

A genetic variant of human serum albumin (alloalbumin) exhibited atypical electrophoretic mobility and chromatographic behavior apparently because of the effect of a point substitution on the molecular conformation. Three forms of albumin were isolated by DEAE HPLC chromatography: normal albumin, and two variant forms V1 and V2. The point substitution (Asp-63-->Asn) generated a canonical tripeptide acceptor sequence for glycosylation with an N-linked oligosaccharide (Asn-Lys-Ser). Neuraminidase digestion followed by electrophoresis showed that the V2 variant form was glycosylated and the V1 form was not. Time-of-flight mass spectrometry yielded a molecular weight of about 2000 for the carbohydrate. Structural analysis of the carbohydrate was done by chromatographic comparison of the pyridylaminated derivatives with standards and was confirmed by proton NMR of the three pronase glycopeptides and of the pyridylaminated oligosaccharide. The oligosaccharide had a complex biantennary structure with two sialic acid residues. In normal albumin Asp-63 is exposed and is adjacent to the first disulfide bond, Cys-62-->Cys-53. The apparent effect on molecular conformation resulting in incomplete glycosylation and atypical electrophoretic mobility suggests that glycosylation may interfere with disulfide bond formation at this site.

Amino Acid Sequence↗

HYS2, an essential gene required for DNA replication in Saccharomyces cerevisiae.

To investigate cell cycle regulation at the S or G2 phase in Saccharomyces cerevisiae, we have isolated mutants displaying supersensitivity to hydroxyurea (HU), a chemical that inhibits DNA replication. Such mutants, which we have named hydroxyurea sensitive (hys), defined four linkage groups and we characterized the hys2 mutation in this study. The hys2-1 mutant displays temperature sensitive growth and a constellation of phenotypes indicating defective DNA metabolism. At the restrictive temperature, hys2-1 cells arrest as large budded cells with a single nucleus at the neck of the bud and a short spindle. The hys2-1 mutant exhibits increased rates of chromosome loss and recombination. Additionally, hys2-1 appears to accumulate incompletely replicated DNA that can be detected by a pulse field electrophoresis assay. Finally, deletion of RAD9 in a hys2-1 strain decreases the percentage of arrested cells, suggesting that an intact RAD9-checkpoint is required for the cell cycle arrest in hys2-1 cells. HYS2 encodes a 55 kDa protein that is essential for viability at all temperatures. Taken together, these data suggest that Hys2 plays a role in DNA replication.

Base Sequence↗

A novel 6Fe (2 x [3Fe-4S]) ferredoxin from Mycobacterium smegmatis.

A novel ferredoxin was purified from Mycobacterium smegmatis by a series of hydrophobic chromatographies in the presence of high concentrations of ammonium sulfate and sodium chloride. The ferredoxin exhibited the same peptide map and N-terminal amino acid sequence as the known 7Fe ferredoxin from the same bacterium. On the other hand, this ferredoxin was found to contain approximately 6 Fe/mol ferredoxin and was also shown to contain only [3Fe-4S] clusters by resonance Raman spectroscopy, indicating that it is a novel 6Fe ferredoxin which contains two [3Fe-4S] clusters.

Electron Spin Resonance Spectroscopy↗

Effects of Ringer's acetate solution during transient hepatic inflow occlusion in rabbits.

This study aimed to clarify the difference in the effects of sodium acetate and sodium lactate administration on hepatic energy metabolism during hepatic warm ischemia and reperfusion. In the first experiment, Ringer's acetate (AR) or Ringer's lactate (LR) solutions were administered intravenously during 20 min of hepatic inflow occlusion in rabbits. Blood gas analyses and measurements of blood pressure, pyruvate, lactate, and ketone body concentrations in arterial blood were performed until 30 min of reperfusion. Hepatic tissue adenine nucleotide concentrations were determined at the end of the experiment. With AR administration, the plasma pyruvate level and the ratio of pyruvate to lactate were significantly elevated during reperfusion. Hepatic energy charge at 30 min of reperfusion improved significantly (P < 0.001) with AR compared with LR administration. Plasma ketone body concentrations decreased markedly with LR administration, but were maintained with AR administration. In the second experiment, intravenous administration of AR or LR, hepatic ischemia, and reperfusion were similarly performed in rabbits. Ketone body concentrations were determined in arterial and vena cava blood at 0 min, 20 min of inflow occlusion, and 30 min of reperfusion. With AR administration, a large arteriovenous difference in ketone body concentrations was observed, indicating utilization in the peripheral tissues. With LR administration, no arteriovenous difference was observed. Availability of energy substrate in peripheral tissues by administration of AR is thought to decrease the metabolic load to the liver and to improve hepatic energy status during reperfusion.

Adenine Nucleotides↗

Phosphatidylinositol hydrolysis is involved in production of Ca(2+)-dependent currents, but not non-selective cation currents, by muscarine in chromaffin cells.

Whether phosphatidylinositol hydrolysis and a subsequent Ca2+ mobilization are responsible for muscarine-induced transient outward currents (IO) and non-selective cation currents (INS) in the guinea-pig chromaffin cell was investigated using the perforated patch method. IO, but not INS, failed to be reproduced in Ca(2+)-free solution and was markedly reduced by prior exposure to caffeine under Ca(2+)-free conditions or by addition to normal solution of cyclopiazonic acid (CPA), a Ca2+ ATPase inhibitor. Application of CPA in Ca(2+)-free solution, however, suppressed INS by about 50% in 73% of the cells tested. Bath application of 1.5 mM neomycin, a phospholipase C inhibitor, induced the time-dependent decline of IO with near abolition at 20 min or less, whereas it produced a time-independent decrease of INS and an inwardly rectifying K+ current. INS in the presence or absence of neomycin was well fitted to rectangular hyperbolas with the same ED50 of 2.17 microM, but with a 33% smaller maximum amplitude in the former, indicating a non-competitive inhibition by neomycin. We conclude that, while phosphatidylinositol hydrolysis mediates the production of IO, it does not mediate that of INS by muscarine.

Animals↗