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Biomedical subjects

S Yoshida

Publications and source records attributed to S Yoshida.

At least 217 records · Page 12Linked to original sources
PubMed2001

Cell type-specific localization of sphingosine kinase 1a in human tissues.

Cell type-specific localization of sphingosine kinase 1a (SPHK1a) in tissues was analyzed with a rabbit polyclonal antibody against the 16 C-terminal amino acids derived from the recently reported mouse cDNA sequence of SPHK1a. This antibody (anti-SPHK1a antibody) can react specifically with SPHK1a of mouse, rat, and human tissues. Utilizing its crossreactivity to human SPHK1a, the cell-specific localization of SPHK1a in human tissues was histochemically examined. Strong positive staining for SPHK1a was observed in the white matter in the cerebrum and cerebellum, the red nucleus and cerebral peduncle in the midbrain, the uriniferous tubules in the kidney, the endothelial cells in vessels of various organs, and in megakaryocytes and platelets. The lining cells of sinusoids in the liver and splenic cords in the spleen showed moderate staining. Columnar epithelia in the intestine and Leydig's cells in the testis showed weak staining patterns. In addition, TPA-treated HEL cells, a human leukemia cell line, showed a megakaryocytic phenotype accompanied with increases in immunostaining of both SPHK1a and SPHK enzyme activity, suggesting that SPHK1a may be a novel marker of megakaryocytic differentiation and that this antibody is also useful for in vitro study of differentiation models.(J Histochem Cytochem 49:845-855, 2001)

Animals↗
PubMed2001

Nuclear localization of neutral sphingomyelinase 1: biochemical and immunocytochemical analyses.

To examine the intracellular localization of neutral sphingomyelinase 1 (nSMase 1), a rabbit polyclonal antibody was raised against a recombinant form of the enzyme expressed in E. coli. It has been reported that, in rat liver or in ascites hepatoma AH7974, high activity of neutral sphingomyelinase (SMase) is found at the plasma membrane, with a lesser but significant amount in nucleus and cytoplasm. The biochemical properties, dithiothreitol requirement and high salt concentration dependency, of cloned and expressed nSMase 1 resemble those of previously described nuclear neutral SMase of AH7974. The present study was therefore focused on the nuclear localization of this enzyme. Western blotting of subcellular fractions using anti-rat nSMase 1 antibody revealed most nSMase 1 to be associated with the nuclei and some with microsomes, but not with plasma membranes. Consistently, neutral SMase activity in nuclear extract was immunoprecipitated by the antibody, while that of plasma membranes was not. The results indicate that nSMase 1 mainly resides in the nucleus and may thus differ from neutral SMase in plasma membrane. On gel-filtration column chromatography of nuclear extract, the profile of neutral SMase activity corresponded well with immunoreactive protein bands on western blotting, suggesting that a large part of nuclear neutral SMase may be nSMase 1. Removal of the nuclear envelope by treatment with Triton X-100 did not significantly decrease the amount of nuclear nSMase 1, and western blotting of subnuclear fractions (i.e. nuclear envelope, chromatin, and nuclear matrix) revealed nSMase 1 signal exclusively in the nuclear matrix. Immunocytochemistry with AH7974, as well as rat fibroblast cell line 3Y1, demonstrated nSMase 1 to be localized mainly in the nucleus, with some in the cytoplasm. Moreover, immuno-electron microscopy clearly showed the signal of nSMase 1 to be more dense in the nucleus than in the cytoplasm of AH7974.

Animals↗
PubMed2001

Galactosyldiacylglycerol, a mammalian DNA polymerase alpha-specific inhibitor from a sea alga, Petalonia bingbamiae.

The glycolipid galactosyldiacylglycerol (GDG), containing C16:0 and C18:1 fatty acids, was isolated from the sea alga Petalonia bingbamiae as a potent inhibitor of the activities of mammalian DNA polymerase alpha (pol. alpha). GDG, however, had no effect on pol. alpha from a fish or a higher plant. The inhibition of pol. alpha by GDG was dose-dependent with an IC50 value of 54 microM. The compound did not influence the activities of other replicative DNA polymerases such as mammalian pol. delta, or repair-related enzymes such as mammalian pol. beta. GDG also did not influence the activities of prokaryotic DNA polymerases such as the Klenow fragment of DNA polymerase I, T4 DNA polymerase, Taq DNA polymerase, DNA polymerases from the higher plant, cauliflower, or DNA metabolic enzymes such as calf thymus terminal deoxynucleotidyl transferase, human immunodeficiency virus type 1 reverse transcriptase and deoxyribonuclease 1. Kinetic analysis of the compound showed that pol. alpha was non-competitively inhibited with respect to both the DNA template and the nucleotide substrate. In this study, we demonstrated the structure-function relationship in the selective inhibition of pol. alpha by the glycolipid group.

Animals↗
PubMed2001

Cardiac sympathetic dysfunction in an athlete's heart detected by 123I-metaiodobenzylguanidine scintigraphy.

The athlete's heart is commonly characterized by an increase in left ventricular mass because of an increase in the left ventricular diastolic cavity dimensions or wall thickness or both. Endurance exercise also induces numerous cardiovascular adaptations, including increased vagal tone. However, the sympathetic function has not yet been precisely elucidated, so the present study evaluated cardiac sympathetic nerve function from metaiodobenzylguanidine (MIBG) images obtained 15 and 180 min after the injection of 123I-MIBG at a dose of 111 MBq. The ratio of heart/mediastinum count (H/M) and the washout rates of 123I-MIBG (WR) were calculated in 25 consecutive patients who were athletes (aged 52+/-13 years) and 23 normal subjects. There was a significant difference in the H/M between the athletic and normal hearts (2.3+/-0.3 vs 2.6+/-0.3, p<0.01, Scheffe's test). An increased WR was observed in the athletes group when compared with the normal group (34+/-4 vs 28+/-3, p<0.01), and there was a significant correlation between WR and the left ventricular mass index (r=0.578, p<0.01). Prolonged exercise training may alter cardiac sympathetic nerve function, which can be detected by MIBG imaging.

3-Iodobenzylguanidine↗
PubMed2001

Inhibition of concanavalin A-induced mice hepatitis by coumarin derivatives.

The effects of coumarin derivatives, osthole, imperatorin, Pd-Ia, Pd-II and Pd-III, on mice concanavalin A (Con A) (0.2 mg/mouse, i.v.)-induced hepatitis were studied. At the dose of 200 mg/kg (i.p.), these coumarins inhibited more than 90% of the Con A-induced elevation of plasma alanine aminotransferase activity, but glycyrrhizin (200 mg/kg, i.p.) caused only 45% inhibition. At the dose of 100 mg/kg (i.p.), osthole produced the strongest inhibition among these coumarins. The inhibitory activity of osthole is lost when its 7-methoxy group is replaced by a 7-hydroxy group to form osthenol. The present results showed that coumarin derivatives inhibited Con A-induced hepatitis, with osthole being the most inhibitory.

Alanine Transaminase↗
PubMed2001

Activation of a potassium conductance by extracellular alkaline pH in oocytes of Xenopus laevis.

Electrophysiological properties of Xenopus oocytes exposed to alkaline extracellular pH (pHo) were investigated by measuring whole-cell currents using the two-electrode voltage-clamp method. Alkaline pHo (8.5-10.5) elicited an outward current in a pHo-dependent manner with a concomitant increase in the membrane conductance. This outward-current response was dependent on K+ because it was suppressed by a K+ channel blocker tetraethylammonium+ (20 mM), and the reversal potential of the response was in good agreement with the Nernst equation for K+. The response was not affected by pretreatment of oocytes with the acetoxymethyl ester of bis-(o-aminophenoxy)-ethane-N,N,N',N'-tetraacetic acid (10,uM), a membrane-permeant intracellular Ca2+ chelator, but it was augmented by forskolin (0.4 microM), a stimulant of adenylate cyclase. The outward-current response originates in the oocyte but not in the surrounding follicle cells because the current could still be evoked when follicle cells were removed by collagenase or when gap junctions connecting the oocyte membrane and follicle cells were blocked by 1-octanol (1 mM). It is concluded that the outward current elicited by alkaline pHo in Xenopus oocytes is dependent on the activation of K+ channels via the cAMP pathway and that the outward current originates in the oocyte rather than the surrounding follicle cells.

Animals↗
PubMed2001

Spiral CT venography of the lower extremities by injection via an arm vein in patients with leg swelling.

The purpose of this prospective study was to assess the role of spiral CT venography (CTV) via an arm vein injection in the detection of causes of leg swelling. 42 consecutive patients with leg swelling were studied with indirect spiral CTV and ultrasound (US). CT parameters were as follows: 5 mm beam collimation; 7-10 mm s(-1) table speed; and 2-3 mm reconstruction. Two consecutive spiral scans with a 40 s exposure time were performed from the pelvis to the knee. One bolus of 150 ml non-ionic contrast medium was injected at a rate of 3 ml s(-1) by a power injector via an arm vein. The delay times to the first and second scans were 120 s and 180 s, respectively. Spiral CTV demonstrated not only deep vein thrombosis (DVT) (n=12) but also other abnormalities (n=25). US showed DVT (n=10) and some other abnormalities (n=5). The sensitivity and specificity of spiral CTV for femoropopliteal DVT, as compared with US, were both 100%. Two cases of DVT in the left common-external iliac vein (iliac vein compression syndrome) detected by spiral CTV were not confirmed by US. We were able to evaluate DVT above the knee with this method. Indirect spiral CTV showed promise for the diagnosis of DVT and other soft tissue diseases in patients with leg swelling.

Adult↗
PubMed2001

Regulation of the localization of Dbf2 and mob1 during cell division of saccharomyces cerevisiae.

The mitotic exit network (MEN) governs Cdk inactivation. In budding yeast, MEN consists of the protein phosphatase Cdc14, the ras-like GTPase Tem1, protein kinases Cdc15, Cdc5, Dbf2 and Dbf2-binding protein Mob1. Tem1, Dbf2, Cdc5 and Cdc15 have been reported to be localized at the spindle pole body (SPB). Here we report changes of the localization of Dbf2 and Mob1 during cell division. Dbf2 and Mob1 localize to the SPBs in anaphase and then moves to the bud neck, just prior to actin ring assembly, consistent with their role in cytokinesis. The neck localization, but not SPB localization, of Dbf2 was inhibited by the Bub2 spindle checkpoint. Cdc14 is the downstream target of Dbf2 in Cdk inactivation, but we found that the neck localization of DbP2 and Mob1 was dependent on the Cdc14 activity, suggesting that Dbf2 and Mob1 function in cytokinesis at the end of the mitotic signaling cascade.

Cell Cycle Proteins↗
PubMed2001

Determination of radionuclides produced by neutrons in heavily exposed workers of the JCO criticality accident in Tokai-mura for estimating an individual's neutron fluence.

In the Tokai-mura criticality accident, three workers were heavily exposed. Biological materials, such as blood, urine, vomit and hair, were collected from the workers and analyzed for radioactivities, produced by the neutron irradiation. Activation products. such as 24Na, -K and 82Br, were found in these materials by gamma-ray spectrometry. The radionuclide of the highest activity observed in biological materials was 24Na, e.g. the concentrations of this nuclide in the blood samples from the three patients at the accident time were 169, 92 and 23 Bq/ml, respectively. The concentrations of stable sodium in the same samples were determined by ICP-AES to obtain specific activities of 24Na (concentration ratio between the produced 24Na and stable 23Na), which are essential for estimating the neutron fluences and radiation doses. The specific activities of 24Na obtained for the three patients through the blood analysis were 8.2 x 10(4),4.3 x 10(4) and 1.2 x 10(4) Bq24Na/g23Na. Based on these values, individual's neutron fluences were estimated to be 5.7 x 10(11), 3.0 x 10(-1) and 0.85 x 10(11) cm(-2), respectively.

Adult↗
PubMed2001

Cytokinin-induced gene expression in cultured green cells of Nicotiana tabacum identified by fluorescent differential display.

The cell growth and plastid development of cultured green tobacco cells were maintained by the phytohormone cytokinin. After subculture into cytokinin-free medium, when cytokinin treatment was resumed, physiological changes induced by cytokinin were analyzed. Changes in chlorophyll biosynthesis and photosynthetic gene expression were observed 1 week after cytokinin induction, and changes in cell growth were observed 2 weeks after cytokinin induction. Two cytokinin-induced genes (cig) were isolated from these cells using the fluorescent differential display technique. Northern analysis confirmed that expression of these cig was induced by both natural and synthetic cytokinins. The expression of cig1 was also induced by abscisic acid, and its cDNA sequence was similar to the proline dehydrogenase gene. The expression of cig2 is specific to cytokinin and is not induced by other phytohormones. The amino acid sequence encoded by cig2 is similar to the GDP/GTP exchange factor eIF2B, which regulates translation initiation. The expression of these cig suggests a complex induction system involving cytokinin and other phytohormones.

Amino Acid Sequence↗
PubMed2001

Biological evaluation of 5-substituted pyrimidine derivatives as inhibitors of brassinosteroid biosynthesis.

A series of 5-substituted pyrimidine derivatives was synthesized, and their ability to inhibit brassinosteroid biosynthesis was tested. The biological activity of these compounds was evaluated by the cress stem elongation method. Among the synthesized compounds, alpha-(4-chlorophenyl)-alpha-phenyl-5-pyrimidinemethanol (DPPM 4) exhibited potent inhibitory activity for retarding cress stem elongation in the light. This inhibition was reversed by the application of 10 nM brassinolide, but not by 1 microM GA3. DPPM 4 also affected Arabidopsis growth in the dark. DPPM 4-treated Arabidopsis had phenotypes like those of brassinosteroid-deficient mutants, with short hypocotyls and open cotyledons, in the dark. These biological changes were restored by the co-application of 10 nM brassinolide, but not by 1 microM GA3, suggesting that the primary site of action of DPPM 4 was the brassinosteroid biosynthetic pathway.

Phenotype↗
PubMed2001

The polymorphism in the caudal-related homeodomain protein Cdx-2 binding element in the human vitamin D receptor gene.

The major physiological activity of 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] is the regulation of calcium absorption in the small intestine, and the level of vitamin D receptor (VDR) is an important factor in this regulation. In a previous study, we indicated-that the caudal-related homeodomain Cdx-2 played an important role in the intestine-specific transcription of the human VDR gene. In this study, the polymorphism was identified in the core sequence 5'-ATAAAAACTTAT-3' in the Cdx-2 binding site in the VDR gene promoter. In 261 Japanese women with genotyped VDR polymorphisms, 48 were genotype Cdx-A (adenine at -3731 nucleotides [nt] relative to the transcription start site of human VDR gene 5-ATAAAAACTTAT), 82 were genotype Cdx-G (guanine at -3731 nt, 5'-GTAAAAACTTAT-3'), and 131 were genotype Cdx-A/G (heterozygote). In postmenopausal Japanese women, the bone mineral density (BMD) in the lumbar spine (L2-L4) with the Cdx-G homozygote was 12% lower than that with the Cdx-A homozygote (p < 0.05). In electrophoretic gel mobility shift assay (EMSA), the oligonucleotide with Cdx-G allele markedly decreased the binding to Cdx-2 compared with that in the Cdx-A allele. The transcriptional activity of the VDR promoter with Cdx-G allele was decreased to 70% of the Cdx-A allele. In addition, in the herpes simplex virus thymidine kinase promoter, the Cdx-2 binding element with the G allele showed significantly lower transcriptional activity than that of the A allele. Thus, the polymorphism in the Cdx-2 binding site of the VDR gene (Cdx-polymorphism) would affect the expression of VDR in the small intestine. In addition, this polymorphism may modulate BMD in postmenopausal Japanese women.

Adult↗
PubMed2001

Increases of calcium and magnesium and decreases of phosphorus and iron with aging in human uterine tubes.

To elucidate compositional changes of the uterine tube by aging, the authors studied age-related changes of elements in human uterine tubes by inductively coupled plasma-atomic emission spectrometry. The uterine tubes were resected postmortem or surgically removed from patients with uterine myoma. It was found that the contents of calcium and magnesium increased progressively with aging in uterine tubes, whereas the contents of phosphorus and iron decreased gradually with aging. The sulfur content of uterine tubes remained constant and independent of aging. Regarding relationships between elements, significant relationships were found between calcium and magnesium contents, between phosphorus and iron contents, between phosphorus and sulfur contents, and between phosphorus and sodium contents in human uterine tubes.

Aged↗
PubMed2001

Age-related changes in the concentrations of major and trace elements in the brain of rats and mice.

Age-related changes in the concentrations of constituent elements in the brains of rats and mice 1 wk to 24 mo old were determined with inductively coupled plasma-mass spectrometry (ICP-MS) and inductively coupled plasma-atomic emission spectrometry (ICP-AES). Seventeen elements could be determined with reasonable accuracy and reproducibility. They were P, K, Na, Mg, Ca, Fe, Zn, Cu, Rb, Al, Mn, Sr, Mo, Co, Pb, Cs, and Cd in order of concentrations in the adult rat brains. In these elements, six major elements (P, K, Na, Fe, Mg, Ca) were determined with ICP-AES and the others with ICP-MS. The concentrations of each element and the pattern of age-related changes were similar between the rat and mouse brains. The elements of which concentrations decreased with aging were K and Rb. On the other hand, the concentrations of some metal elements, including Fe, Cu, Sr, and Co, appeared to increase with growth and aging. The concentrations of other elements were relatively constant throughout the age examined.

Aging↗
PubMed2001

Effects of ionic valency of interacting metal elements in ion uptake by carrot (Daucas carota cv. U.S. harumakigosun).

Interaction of elements in the course of element uptake by carrot (Daucas carota cv. U.S. harumakigosun) exerted by the addition of elements, such as Rb, Zn, and Al, was investigated. For the purpose of precise evaluation of uptake behavior, the simultaneous determination of absorption of Na, Be, Sr, Mn, Co, Zn, Ce, Pm, and Gd was conducted by the multitracer technique. For root uptakes, Al exhibited its influence on the uptake of essential elements and on the uptake of toxic or unbeneficial ones, presumably as a result of the large electric valency that caused cell membrane disintegrity. On the other hand, Zn as a divalent cation only affected the uptake of essential and beneficial elements. Rubidium, which is a monovalent cation, did not exhibit any effect on the uptake of other ions. Concerning shoot uptakes, inhibition by Zn and Al, but not by Rb, was observed for the uptake of Sr, Mn, Co, and Zn. From the present investigation, it is suggested that there exists an interaction between added ions and the elements taken into plants and that the degree of interaction increases in the increasing order of ionic valency: M+ (Rb), M2+ (Zn), and M3+ (Al).

Aluminum↗
PubMed2001

Intussusception during enteral nutrition: a case report.

We report a case of intussusception during enteral nutrition after esophagectomy. The case was a 60-year-old patient who underwent subtotal esophagectomy via the right thoraco-abdominal approach, reconstructed with esophagogastrostomy. He underwent tube feeding through gastrostomy after surgery. The tip of the feeding tube was emplaced in the jejunum. He complained of intermittent abdominal pain and the drainage volume through the gastric decompression tube was increased on the 2nd week from surgery. There was no abnormal finding on abdominal CT or or radiography of the digestive tract, and so a diagnosis of intussusception was made and laparotomy was performed. There was descending intussusception with the jejunum where the feeding tube was emplaced. Enterectomy was not necessary in this case. Adult intussusception is rare. The surgery is sometimes the cause for intussusception. Another cause is use of a long intestinal tube. Since enteral nutrition has priority for nutritional support after esophagectomy, it is important to make an early decision whether surgical intervention is required or not, using abdominal examination by CT, ultrasound and contrast radiography, when bowel obstruction occurs during enteral nutrition through a feeding tube.

Enteral Nutrition↗
PubMed2001

Characterization of glycans in a lactoferrin isoform, lactoferrin-a.

The presence of glycan at Asn-281 in bovine lactoferrin-a, which has a higher molecular weight than regular lactoferrin-b, was found in our previous study. The present work was performed to clarify the structures of the glycans linked to the five N-glycosylation sites in lactoferrin-a and to compare them with those of glycans linked to lactoferrin-b. In lactoferrin-a, the glycans linked to Asn-233 and Asn-545 were of the high-mannose type, whereas those present at Asn-368 and Asn-476 were complex-type ones. These glycans possessed heterogeneous structures. A comparative study of the glycans on bovine lactoferrin-a and bovine lactoferrin-b by HPLC showed that the structures of the glycans linked to Asn-368, Asn-476, and Asn-545 were very similar, the exception being the glycan linked to Asn-233. In addition, analysis of the structure of the glycan bound to Asn-281 present only in lactoferrin-a showed it possessed the heterogeneous structure of a complex-type glycan in which the structures Man3GlcNAc2, Man3GlcNAc4, Man3GlcNAc4Fuc are suggested to be present based on HPLC retention times only.

Animals↗