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Biomedical subjects

H Yamaguchi

Publications and source records attributed to H Yamaguchi.

At least 721 records · Page 40Linked to original sources

Flow cytometric analysis using lipophilic dye PKH-2 for adhesion of Vibrio cholerae to Intestine 407 cells.

A comparative study of indirect and direct flow cytometric analysis for adherence of Vibrio cholerae to Intestine 407 cells was performed. The direct flow cytometric analysis employed the lipophilic dye PKH-2. It was concluded that direct flow cytometry using the lipophilic dye PKH-2 is useful and convenient for analyzing bacterium-host cell interactions, since it does not require any specific antibody as the first antibody.

Bacterial Adhesion↗

Basic fibroblast growth factor as a candidate tumor marker for renal cell carcinoma.

The present investigation was conducted to determine serum levels of basic fibroblast growth factor (FGF) by enzyme immunoassay in patients with various urogenital tumors. Renal cell carcinoma had a higher tendency (28 of 52, 53.8%) toward increased serum levels of basic FGF than any of the other urogenital tumors, and increased serum basic FGF was detected more frequently in patients with advanced renal cell carcinoma. Analysis of histological pattern indicated that renal cell carcinoma with a solid or tubular component is more likely to produce basic FGF. However, no significant difference was seen between the percentage of clear cell type tumor patients with increased serum basic FGF (50.0%) and the percentage of granular cell type tumor patients with increased serum basic FGF (66.7%). Five of 8 patients with renal cell carcinoma who underwent selective renal venous sampling before nephrectomy showed increased serum basic FGF in the renal vein from the affected kidney. After resection of the affected kidney to remove the cancer, serum basic FGF disappeared within 2 weeks. However, residual huge tumor or postoperative disease prolonged the increased levels of basic FGF in 2 patients, indicating that basic FGF is produced from and secreted by tumor tissue itself. These findings suggest that serum basic FGF can be useful in the diagnosis, and in evaluating the prognosis, of patients with renal cell carcinoma.

Biomarkers, Tumor↗

Simultaneous DNA typing of human platelet antigens 2, 3 and 4 by an allele-specific PCR method.

We developed an allele-specific polymerase chain reaction (ASPCR) method using originally designed primers to determine the genotype of the human platelet antigens (HPAs) 2, 3 and 4 in parallel. The results were compared with those obtained by PCR restriction fragment length polymorphism and the mixed passive hemagglutination test. Seventy-three individuals were tested and the ASPCR results were in good agreement with those determined by the other two methods. This method enables the genotyping of HPA-2, -3 and -4 in parallel without the use of platelets, platelet-specific alloantibodies or restriction enzymes.

Alleles↗

Changes in apolipoproteins during the acute phase of Kawasaki disease.

To determine disturbances of lipid metabolism in patients with Kawasaki disease, we investigated changes in the serum levels of apolipoproteins and serum lipids. Results were as follows. Total serum cholesterol (TC) and high-density lipoprotein cholesterol (HDL-C) decreased during the early stage of Kawasaki disease. The apo A-I and A-II levels were low until the 2nd week of illness. Apo B decreased during the 1st week of illness, but rose slightly during the 2nd and 3rd weeks. The apo B/apo A-I ratio and the apo B/TC ratio were high in the early stages of illness. The group of patients with coronary artery lesions showed low levels of TC and HDL-C, and low levels of apo A-I, A-II and B during the early stages, compared with the group without coronary artery lesions. The apo B/TC ratio was significantly higher in the patients with coronary artery lesions during the 2nd week of illness. Our findings suggest an association between changes in serum lipids and apolipoproteins and coronary artery involvement in Kawasaki disease. These abnormalities may indicate the presence of early coronary arteriosclerosis.

Acute Disease↗

Clinical and electrophysiological characteristics of atrial standstill.

To clarify the clinical and electrophysiological characteristics of atrial standstill (AS) we studied 11 patients (7 males and 4 females), whose average age was 62 years and who were followed over a period of 4-179 months. Underlying heart disease was present in nine patients and two cases were idiopathic. Major clinical symptoms in the 11 cases included Adams-Stokes attacks, and dyspnea on exertion. In the standard 12-lead ECGs obtained on admission, the P wave was absent in six cases. Atrial flutter (AF) was noted in 3, atrial fibrillation (Af) in 1, and multifocal atrial tachycardia in 1. In some cases, the ECG initially showed AF or Af, and was transformed after several years into ectopic atrial tachycardia or an ectopic atrial rhythm with a markedly decreased amplitude of the P wave. Finally, the P wave disappeared over a prolonged period. When intracardiac mapping was performed, the atrial electrograms tended to diminish at the site of high, mid-lateral right atrium (RA). Electrograms were remained present in the vicinity of the tricuspid valve (TV) annulus. A repeated mapping and pacing study conducted in two patients revealed that the "silent" area spread toward the lower site of RA. During the average follow-up period of 64 months, four patients died. The interval until death in one patient with myocarditis was 6 months, and in another with dilated cardiomyopathy (DCM) it was 8 months. It appears that the atrial muscular lesion starts in the high lateral RA and progresses toward the lower RA, then to the vicinity of the TV annulus. A diffuse and progressive disturbance may occur not only in the atrial muscle, but also in the atrioventricular conduction system in patients with AS who had progressive myocarditis or DCM.

Aged↗

Short-term topical therapy of experimental tinea pedis in guinea pigs with lanoconazole, a new imidazole antimycotic agent.

The therapeutic efficacy of short-term treatment with a 1% cream of lanoconazole, a new imidazole antimycotic agent, in comparison with that of a 1% cream of terbinafine was evaluated in the guinea pig model of tinea pedis. Each agent was topically applied once a day for 3 or 7 consecutive days, starting on day 10 postinfection, and a culture study was conducted on day 5 after the last treatment with each agent. The 1% cream of lanoconazole was as highly effective as the 1% cream of terbinafine in terms of eradicating the fungi from the infected feet.

Administration, Topical↗

Isoproterenol increases peripheral [Ca2+]i and decreases inner [Ca2+]i in single airway smooth muscle cells.

The effects of isoproterenol (Iso) or photolysis of caged adenosine 3',5'-cyclic monophosphate (cAMP) on intracellular Ca2+ concentration ([Ca2+]i) were studied in single airway smooth muscle cells. Changes in [Ca2+]i were measured ratiometrically. In cells loaded with 10 microM fura 2-acetoxymethyl ester (AM), superfusion of Iso (10 microM) increased [Ca2+]i in 20 of 22 cells from 153.2 +/- 21.3 to 252.8 +/- 38.3 nM, and this increase depended on extracellular Ca2+ and was blocked by ryanodine (50 microM). Photolysis of intracellular caged cAMP increased [Ca2+]i by 104.0 +/- 17.3 nM in 20 cells and decreased [Ca2+]i by 49.3 +/- 9.4 nM in 10 cells. With modified confocal microscopy, peripheral [Ca2+]i was increased and inner cytosolic [Ca2+]i was decreased during stimulation with Iso. Iso-induced decreases in [Ca2+]i were also observed with conventional optics in cells loaded with 0.5 microM fura 2-AM. However, in these same cells, Iso increased [Ca2+]i in the presence of low concentrations of ryanodine (1-20 microM). We concluded that Iso decreased [Ca2+]i in the inner cytosol but increased [Ca2+]i in the peripheral cytosol by mechanisms that depended on both extracellular and sarcoplasmic reticulum Ca2+ release channels. Our study suggests that fluorescence from a peripheral cytosol with high [Ca2+]i can sometimes confound the measurement of "cytosolic" [Ca2+]i with conventional optics.

Animals↗

Production and expression of granulocyte- and macrophage-colony-stimulating factors in newborns: their roles in leukocytosis at birth.

We analyzed the production and expression of three colony-stimulating factors (CSFs) in neonates to clarify the mechanism of leukocytosis at birth. Serial blood samples (n = 23) were collected from mothers, cord blood, and from newborn infants on days 1, 5, and 30 after birth. The serum levels of granulocyte-CSF (G-CSF), granulocyte/macrophage/CSF (GM-CSF) and macrophage-CSF (M-CSF) were measured by ELISA. The G-CSF levels on day 1 after birth were significantly higher than those thereafter, and they were also higher in the mothers than those on days 5 and 30 after birth. The GM-CSF levels did not change significantly during the neonatal period. The serum M-CSF levels were higher on postnatal day 1 than at other times, and gradually decreased thereafter. To confirm the production sites of G-CSF and M-CSF, the mRNA for these CSFs in peripheral mononuclear cells (MNCs) from healthy adults, mothers, and cord blood were analyzed by PCR. The expression of G-CSF and GM-CSF mRNA was undetectable in MNCs from adults, mothers, and cord blood, while these cells expressed low levels of M-CSF mRNA. After stimulation with lipopolysaccharide or phorbol myristate acetate, the MNCs expressed high levels of G-CSF and GM-CSF mRNA. The levels of G-CSF PCR products in cord MNCs were lower than those in adult and maternal MNCs. The expression of M-CSF mRNA was virtually unchanged by stimulation. To detect the localization of G-CSF and M-CSF in the placenta and umbilical cord, these tissues were immunocytochemically stained with anti-G-CSF and anti-M-CSF antibodies, G-CSF and M-CSF were expressed in trophoblasts and decidual stromal cells, whereas the umbilical cord did not express these CSFs. Moreover, large amounts of G-CSF and M-CSF were detected in the supernatant of cultured trophoblasts and decidual stromal cells. The expression of G-CSF and M-CSF in these cells was confirmed by PCR. These findings suggested that G-CSF and M-CSF produced in the placenta (trophoblasts and decidual stromal cells) are the major factors that induce leukocytosis in newborn infants at birth.

Base Sequence↗

Properties of Ca(2+)-dependent K+ channels of human gingival fibroblasts.

Cells in the oral cavity are normally exposed to different temperatures. Ion transport systems are influenced by temperature in other tissues: In particular, changes in intracellular K+ ion can affect cell growth and synthesis of macromolecules. The purpose of this investigation was to identify K+ channels in human gingival fibroblast cells and analyze the effect of temperature on their K+ conduction properties. Ca(2+)-dependent K+ channels with a large conductance (125 pS in symmetrical K(+)-rich solutions) were identified in human gingival fibroblasts and studied by the patch-clamp technique. The open probability of the channels varied with membrane potential between +40 and -100 mV. When the bath temperature was decreased from 40 to 4 degrees C, channel conductance was reduced, but the mean open time of the channels was increased. The activation energies for the conductance and the reciprocal of the mean open time were estimated to be 9.1 and 22.9 kJ/mol, respectively. These values are lower than those reported for these and other types of channels in cells from other tissues. The open probability of the channels was nearly constant in the temperature range studied. These results suggest that the properties of Ca(2+)-dependent K+ channels of gingival fibroblasts remain relatively unchanged when the cells are exposed to a wide range of temperatures.

Body Temperature↗

Inhibitory effect of sodium 5,6-benzylidene ascorbate (SBA) on the elevation of melanin biosynthesis induced by ultraviolet-A (UV-A) light in cultured B-16 melanoma cells.

Sodium 5,6-benzylidene ascorbate (SBA) is a conjugate of ascorbic acid (Asc) with benzyaldehyde. It has been found that the antioxidant activity of SBA is more stable and has a longer lifetime in living cells and organs than Asc. In this study, we investigated the effect of SBA on the induction of melanin in cultured melanoma (B-16) cells irradiated by UV-A. Melanin content of B-16 cells was significantly increased by UV-A irradiation. The induction was abolished by mannitol and particularly by superoxide dismutase, suggesting the involvement of O2- in the biosynthesis of melanin in cultured melanoma cells. This was theorized by the fact that the induction was also observed in B-16 cells treated with superoxide anion radicals chemically generated in the hypoxanthine/xanthine oxidase-reaction system, instead of UV-A irradiation. The induction of melanin caused by UV-A irradiation was suppressed by SBA in a dose-dependent manner. To elucidate the mechanism of this suppressive effect, the scavenging activity against O2-, and the inhibitory effect of SBA on tyrosinase activity were examined. ESR spectrometric analysis showed that SBA strongly scavenged O2-, and the presence of SBA in the medium remarkably inhibited the tyrosinase activity in cultured B-16 melanoma cells. It can be concluded that SBA effectively inhibits the melanin biosynthesis in B-16 melanoma cells induced by reactive oxygen species (ROS) generated by UV-A irradiation via tyrosinase.

Antioxidants↗

Changes of conductivity in patients with second- or third-degree atrioventricular block after pacemaker implantation.

UNLABELLED: The purpose of this study was to examine the long-term course of conductivity in patients with second- or third-degree atrioventricular (AV) block after pacemaker implantation. Fifty-four patients (30 males and 24 females, mean age 59.5 years) had a 12-lead electrocardiogram (ECG) recorded prior to pacemaker implantation and again after a follow-up of more than 5 years (mean 108 months). The degree of AV block and the morphology of the QRS complex in the two ECGs were compared. Twenty-eight (85%) of the 33 patients with complete AV block and 15 (71%) of the 21 patients with second-degree AV block before implantation showed complete AV block after the follow-up period. The progression of AV block was recognized only in patients with intra-His (BH) or infra-His (H-V) block. In 29 patients, escape QRS complexes were recorded in both ECGs. A change in the QRS complex was noted in 6 patients (21%), including 2 with new bundle branch block (BBB), 2 with wider BBB, 1 with new left axis deviation, and 1 with a change to another BBB. This change was particularly notable in the H-V block group (44%). IN CONCLUSION: (1) Most patients with complete AV block did not recover AV conductivity after the long-term follow-up. (2) Second-degree BH and H-V block tended to progress to complete AV block.

Adolescent↗

Vasopressin V1-receptor stimulation produces a positive inotropic response without affecting pHi in guinea pig papillary muscles.

Effects of arginine-vasopressin (AVP) on the contractile force, action potential (AP) and intracellular pH (pHi) were studied in isolated guinea pig papillary muscles using conventional and ion-selective microelectrode techniques. AVP increased the developed tension and the resting tension, and these responses were attenuated by the V1-receptor antagonist OPC-21268 (1-(1-[4-(3-acetylaminopropoxy)benzoyl]-4-piperidyl)-3,4-dihydro-2 (1H)- quinolinone). However, AVP failed to affect AP configuration or pHi. These results suggest that AVP produces a positive inotropy by mechanism(s) other than intracellular alkalinization.

Animals↗

Synthesis of novel oligosaccharides from leucrose by an alpha-glucosidase.

An alpha-glucosidase was purified in an electrophoretically pure state from an extract of koji culture of Aspergillus sp. KT-11. This enzyme was found to have a transferring activity when the reaction was done in a high concentration of leucrose at pH 4.5. Two kinds of transfer products, fractions I and II, were obtained from leucrose by the enzyme and they were identified as [(alpha-D-glucopyranosyl-(1-->6)-alpha-D-glucopyranosyl-(1-->6)-alpha-D- glucopyranosyl-(1-->5)-D-fructopyranose] and [alpha-D-glucopyranosyl-(1-->6)-alpha-D-glucopyranosyl-(1-->5)-D- fructopyranose], respectively. These are considered to be novel oligosaccharides.

Aspergillus↗

Purification and characterization of angiotensin II degradation factor from porcine endothelial cells.

We investigated the degradation of angiotensin II by vascular endothelial cells and smooth muscle cells in vitro. When angiotensin II was incubated with confluent culture of endothelial cells or with serum free conditioned medium of the endothelial cells, angiotensin II was destroyed rapidly. When angiotensin II was incubated with cultured vascular smooth muscle cells or their serum free conditioned medium, degradation was not observed. To identify the angiotensin II degradation factor (ADF), we have purified ADF from the conditioned medium of endothelial cells, by column chromatographies, i.e., hydroxyapatite, ion exchange and gel filtration chromatography. The partially purified ADF had apparent molecular masses of 154 kDa on gel filtration chromatography. Its pH optimum was about 7.0. ADF was inhibited by p-chloromercuribenzoic acid, iodoacetamide and high concentration of EDTA, but not by diisopropyl fluorophosphate, bestatin, amastatin or pepstatin A. Of the synthetic substrates examined, ADF degrades human angiotensin II, [Val5]-angiotensin II and [Asn1, Val5]-angiotensin II. It did not degrade angiotensin I.

Ammonium Sulfate↗

Ectodermal dysplasia syndrome in siblings with true keloids, stenosis of the esophagus after operations for congenital achalasia and renovascular hypertension due to stenosis of renal artery.

Ectodermal dysplasia syndrome (EDS) is a rare hereditary disease, with symptoms brought about by dysplasia of ectodermal tissue (such as skin, teeth, nails, and hair). This report details the cases of two siblings (41 and 43 year old sisters) with autosomal recessive and hydrotic EDS complicated by esophageal achalasia, postoperative stenosis of esophagus, true keloids, renovascular hypertension, incomplete malrotation of the bowel, and demyelination of the brain.

Adult↗

Depression of ATP-induced Ca2+ signalling by high K+ and low Cl- media in human aortic endothelial cells.

We studied the contribution of the Cl- channel as well as K+ channel in the regulation of Ca2+ signalling in fura-2-loaded cultured human aortic endothelial cells. Low Cl- (20 mM) superfusion did not affect the ATP (10 microM)-induced [Ca2+]i increase at the initial peak (control 309 +/- 30 nM (mean +/- SD, n = 17) versus 20 mM [Cl-]o 308 +/- 40 nM (n = 8)) but depressed it at the sustained phase (at 5 min, 170 +/- 29 nM versus 85 +/- 10 nM). Similar selective depression of the sustained phase occurred also in Ca(2+)-free and 140 mM K+ solutions and in the presence of niflumic acid (300 microM), a blocker of the Cl- channel and Ca2+ permeable cation channel. After application of ATP, the increase of [Cl-]o from 20 to 146 mM resulted in a Ca2+ overshoot. Both Cl- and K+ channels play an important role in the regulation of Ca2+ influx presumably by controlling the membrane potential in vascular endothelial cells.

Adenosine Triphosphate↗

Effect of endothelin-1 on the development of hemorrhagic pancreatitis in rats.

BACKGROUND: It has been hypothesized that microcirculatory disturbance plays an important role in the development of severe pancreatitis. In this study we investigated the effects of exogenous endothelin-1 on the development of severe pancreatitis in rats. METHODS: Acute pancreatitis was induced by two intraperitoneal injections of cerulein (10 micrograms/kg body weight). Endothelin-1 was administered via an abdominal aortic catheter as a bolus of 250-750 pmol/kg BW every hour for 4 h. RESULTS: Remarkable morphologic changes in the pancreas, including hemorrhage, and increases in serum amylase level and active elastase content in pancreatic tissue were observed in rats treated with cerulein plus endothelin-1 in a dose-dependent manner 5 h after the first cerulein injection. Local pancreatic blood flow decreased significantly, and microcirculatory disturbances in the pancreas were demonstrated. CONCLUSIONS: These results suggest that endothelin-1 causes pancreatic microcirculatory disturbance and might be a contributing factor in the aggravation of acute pancreatitis.

Acute Disease↗

Smooth muscle phenotypes in developing and atherosclerotic human arteries demonstrated by myosin expression.

Smooth muscle myosin heavy chains (MHC) exist in multiple isoforms. Rabbit smooth muscle contain at least three types of MHC isoforms; SM1 (204 kDa), SM2 (200 kDa) and SMemb (200 kDa). SM1 and SM2 are specific to smooth muscle, but SMemb is a nonmuscle-type MHC abundantly expressed in the embryonic aorta and in activated mesenchymal cells. We previously reported that these three MHC isoforms are differentially expressed in rabbit during normal vascular development and in experimental arteriosclerosis and demonstrated that MHC isoforms are excellent markers for smooth muscle phenotype. In order to clarify the clinical significance of MHC isoforms, this article will focus on the expression of smooth muscle MHC isoforms in normally developing and atherosclerotic human arteries, especially in coronary arteries. We recently isolated and characterized three cDNA clones encoding human SM1, SM2, and SMemb. The expression of SM2 mRNA in the human fetal aorta was significantly lower as compared to SM1 mRNA but the ratio of SM2- to SM1-mRNA was increased after birth. SMemb mRNA in the aorta was decreased after birth. Immunohistologically, SM1 was constitutively positive from the fetal stage to adulthood in the apparently normal media of the aorta and coronary arteries, whereas SM2 was not detected in fetal arteries of early gestational stage. SM2 was recognized in well-differentiated smooth muscle after perinatal stage. In the human aorta or coronary arteries, unlike in rabbit, SMemb was detected even in the adult. Mild diffuse intimal thickening in the major coronary arteries of the young was found to be composed of smooth muscle cells, reacting equally to three antibodies for MHC isoforms. In thickened but non-atheromatous intima, the expression of well-differentiated smooth muscle-specific MHC (SM2) was reduced, especially in the deeper layer. With progression of atherosclerosis, intimal smooth muscle diminished the expression of not only SM2 but also SM1, whereas alpha-smooth muscle actin was well preserved. We conclude from these results that smooth muscle MHC isoforms are important molecular markers for studying human vascular smooth muscle cell differentiation as well as the cellular mechanisms of atherosclerosis.

Amino Acid Sequence↗