Search PubMed⌕ Search

Biomedical subjects

Y Kudo

Publications and source records attributed to Y Kudo.

At least 37 records · Page 2Linked to original sources
PubMed2002

Changes in expression and function of syncytin and its receptor, amino acid transport system B(0) (ASCT2), in human placental choriocarcinoma BeWo cells during syncytialization.

Relative abundance of mRNAs encoding syncytin and its receptor, amino acid transport system B(0), and activity of amino acid transport thought to be through this system have been studied in parallel in a cell model of syncytialization (BeWo cell following forskolin treatment). Relative mRNA abundance (determined by reverse transcription-polymerase chain reaction) for syncytin showed stimulation by forskolin. In contrast, the level of amino acid transporter B(0) mRNA expression was lower in forskolin treated cells. Na(+)-dependent alpha-(methylamino)isobutyric acid insensitive L -alanine transport was similarly decreased significantly in cells treated with forskolin suggesting that there is modulation of cell surface expression of the syncytin receptor associated with syncytialization.

Alanine↗
PubMed2001

Oxidized low-density lipoprotein-binding specificity of Asp-hemolysin from Aspergillus fumigatus.

Oxidized low-density lipoprotein (Ox-LDL) is known to be involved in the generation and progression of atherosclerosis. Ox-LDL has a number of potentially atherogenic effects on vascular cells, including the uncontrolled uptake by scavenger receptors. We have previously shown that Asp-hemolysin binds to Ox-LDL in a concentration-dependent manner. The present study was undertaken to clarify the binding specificity of Asp-hemolysin to Ox-LDL. We examined the binding specificity of Asp-hemolysin to Ox-LDL using several modified lipoproteins and scavenger receptor ligands. Asp-hemolysin bound to Ox-LDL with shorter LDL oxidation times. However, Asp-hemolysin did not bind to the acetylated LDL. The native high-density lipoprotein (n-HDL) and modified HDL (e.g., acetylated HDL, oxidized HDL) also had no Asp-hemolysin binding. Furthermore, inhibitors of the scavenger receptor binding, including maleylated BSA, polyinosinic acid, dextran sulfate and fucoidin, had no effect on the binding of Ox-LDL to Asp-hemolysin. Surface plasmon resonance studies revealed that Ox-LDL binds with high affinity (K(D)=0.63 microg/ml) to Asp-hemolysin. We concluded that Asp-hemolysin is a specific binding protein with a high affinity for Ox-LDL, and its binding specificity is distinct from any receptor for Ox-LDL. The present studies suggest that Asp-hemolysin may bind to Ox-LDL using a mechanism different from the scavenger receptors.

Aspergillus fumigatus↗
PubMed2001

Transient occurrence of 27 kDa heat-shock protein in the terminal tubule cells during postnatal development of the rat submandibular gland.

It has been suggested that the 27 kDa heat-shock protein (Hsp27) plays a role at crucial cellular checkpoints for proliferation, apoptosis, and differentiation. We examined the immunolocalization of Hsp27 in the rat submandibular gland during postnatal development, wherein acinar cells proliferate and differentiate at earlier postnatal periods. At 2 weeks of age, weak Hsp27 immunoreactivity was distributed diffusely over all gland components. At 3 weeks, Hsp27 immunoreactivity disappeared in most parts of the acini and ducts, but was intensely accumulated in a small cell population located in the acinar center. This population was composed mostly of terminal tubule (TT) type I cells. At 4 weeks, the Hsp27-immunopositive cell population in the acinar center was composed primarily of immature (type II) acinar cells, partly of immature (granulated) intercalated duct (ID) cells, and occasionally of apoptotic cells. After 5 weeks, all acinar components became mature and were no longer immunoreactive for Hsp27. When acinar cell differentiation was accelerated by administration of isoproterenol to 3-week-old rats for 7 days, the number of Hsp27-positive cells was significantly lower than in the control gland at 4 weeks, confirming that Hsp27 expression is downregulated in mature acinar cells. These results suggest that at around 3-4 weeks in postnatal development, the centroacinar TT cells stop proliferating and begin to differentiate into acinar and ID cells, and occasionally undergo apoptosis. Hsp27 is transiently expressed in the centroacinar TT cells during this critical period, and thus may play a role in their differentiation into the immediate descendants.

Animals↗
PubMed2001

Functional and molecular characterization of a peptide transporter in the rat PC12 neuroendocrine cell line.

We have studied functional properties of peptide transport in the pheochromocytoma neuroendocrine cell line from rat. The neutral peptide D-Phe-L-Ala (resistant to hydrolysis) is a good substrate for uptake into these cells. Transport is substantially inhibited by diethylpyrocarbonate pretreatment and is stimulated by external acidification. It is sodium-independent and, unexpectedly, insensitive to membrane potential. Peptide uptake is inhibited by a wide variety of other di- and tripeptides but not by amino acids. The neuropeptide kyotorphin (opioid dipeptide (L-Tyr-L-Arg)) inhibits uptake of labelled peptide and trans-stimulates efflux showing that it is a transported substrate. These findings are discussed in relation to the molecular basis and physiological role of this transport system.

Animals↗
PubMed2001

Establishment of CHO cell lines expressing four N-methyl-D-aspartate receptor subtypes and characterization of a novel antagonist PPDC.

To develop an assay system that allows the N-methyl-D-aspartate (NMDA) receptor subtype-selective antagonistic potency of drugs, we have established Chinese hamster ovary cell lines expressing the four NMDA receptor subtypes (GluRepsilon1/zeta1-GluRepsilon4/zeta1) heat-indelibly. Using these clonal cells, we found that a novel antagonist, (1S,2R)-1-phenyl-2[(S)-1-aminopropyl]-N,N-diethylcyclopropanecarboxamide, was less selective for the GluRepsilon1/zeta1: the IC(50) values for the GluRepsilon1/zeta1-GluRepsilon4/zeta1 were 41.7, 13.3, 12.6 and 11.5 microM, respectively, while two well-known antagonists, DL-2-amino-5-phosphonovaleric acid and ifenprodil, showed the known potency and selectivity for each subtype. Thus, the established clonal cells are of use in characterizing the pharmacological properties of drugs that act on NMDA receptors.

Animals↗
PubMed2001

Analysis of codon usage diversity of bacterial genes with a self-organizing map (SOM): characterization of horizontally transferred genes with emphasis on the E. coli O157 genome.

With increases in the amounts of available DNA sequence data, it has become increasingly important to develop tools for comprehensive systematic analysis and comparison of species-specific characteristics of protein-coding sequences for a wide variety of genomes. In the present study, we used a novel neural-network algorithm, a self-organizing map (SOM), to efficiently and comprehensively analyze codon usage in approximately 60,000 genes from 29 bacterial species simultaneously. This SOM makes it possible to cluster and visualize genes of individual species separately at a much higher resolution than can be obtained with principal component analysis. The organization of the SOM can be explained by the genome G+C% and tRNA compositions of the individual species. We used SOM to examine codon usage heterogeneity in the E. coli O157 genome, which contains 'O157-unique segments' (O-islands), and showed that SOM is a powerful tool for characterization of horizontally transferred genes.

Algorithms↗
PubMed2001

High expression of S-phase kinase-interacting protein 2, human F-box protein, correlates with poor prognosis in oral squamous cell carcinomas.

Reduced expression of p27(Kip1), a cyclin-dependent kinase (Cdk) inhibitor, is frequently found in various cancers, including oral squamous cell carcinoma (OSCC), and is attributable to an enhancement of its degradation. Skp2, an F-box protein necessary for DNA replication, is required for the ubiquitinylation and subsequent degradation of p27(Kip1). In the present study, we examined the expression of Skp2 and its correlation with the expression of p27(Kip1) protein or p27(Kip1) degradation in OSCC. Using immunohistochemistry, we found that high expression of Skp2 was present in 49% of OSCCs and only 20% of epithelial dysplasias. Significantly, high expression of Skp2 was correlated with poor prognosis of OSCC patients. We also found an inverse correlation between the expression of Skp2 and p27 by immunohistochemical analysis. A similar correlation was observed in OSCC cell lines and OSCC tissues by Western blot analysis. Interestingly, OSCC tissues with Skp2 expression had high p27(Kip1) degradation activity. These findings indicate that (a) Skp2 may play an important role for the development of OSCC, (b) Skp2 can be a novel target for OSCC treatment as well as a strong prognostic marker, and (c) the reduction in p27(Kip1) protein may be brought about by enhancement of its degradation mediated by increased levels of Skp2 protein.

Adult↗
PubMed2001

Functional expression of alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid receptors precedes the development of cholinergic phenotype in embryonic rat septal cells in culture.

We examined the development of cholinergic neuronal functions and alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA) responses in cultured embryonic rat septal cells. Choline acetyltransferase activity was increased from 4 to 6 days in culture and reached a plateau at day 8. Acetylcholine release was increased from 6 to 8 days in culture. AMPA-induced increase in intracellular Ca(2+) level was observed at 3 days in culture and most of the AMPA-responsive cells coincided with high-K(+) responsive cells. These results suggest that cholinergic neurons develop their neuronal functions about 8 days under cultured conditions, and functional expression of AMPA receptors precedes the cholinergic functional development.

Acetylcholine↗
PubMed2001

Tryptophan degradation by human placental indoleamine 2,3-dioxygenase regulates lymphocyte proliferation.

1. The physiological importance of human placental indoleamine 2,3-dioxygenase (EC 1.13.11.42), the first and rate-limiting enzyme in tryptophan metabolism, in regulating feto-maternal immunology has been studied. 2. Concentrations were measured in placental villous explant conditioned media of 14 amino acids that are known to be required for lymphocyte proliferation. In the absence of interferon-gamma only tryptophan and threonine were significantly lowered; in the presence of interferon-gamma (known to stimulate indoleamine 2,3-dioxygenase) tryptophan but not threonine depletion was much greater. 3. Peripheral blood mononuclear cell proliferation determined by measuring thymidine incorporation into DNA following culture in the medium previously conditioned by culture of villous explants was markedly reduced when placental indoleamine 2,3-dioxygenase was stimulated with interferon-gamma. Inhibition of placental indoleamine 2,3-dioxygenase by 1-methyl-tryptophan prevented inhibition of thymidine incorporation. Supplementation of the conditioned medium with tryptophan but no other amino acid completely reversed the inhibition of thymidine incorporation. 4. Flow cytometric analysis showed that CD4-positive T lymphocyte division was specifically suppressed by indoleamine 2,3-dioxygenase-mediated tryptophan depletion. This inhibition of T cell proliferation was due to arrest of cell cycle progression. 5. To study the mechanism of tryptophan sensing we examined the ability of 11 L-tryptophan analogues to support lymphocyte proliferation. Only L-tryptophan methyl and ethyl esters were able to stimulate proliferation in tryptophan-free media. Since both of these molecules are readily degraded to tryptophan by intracellular esterases this suggests that the tryptophan sensor is intracellular. 6. Our results show that mechanisms are present in the human placenta which are able to regulate cellular proliferation of the maternal immune system. This mechanism is dependent both on placental indoleamine 2,3-dioxygenase-mediated tryptophan degradation and on tryptophan sensing systems within lymphocytes.

Amino Acids↗
PubMed2001

Characterisation of L-tryptophan transporters in human placenta: a comparison of brush border and basal membrane vesicles.

The mechanisms responsible for L-tryptophan transport at both the maternal- and fetal-facing surfaces of the term placenta have been determined in isolated membrane vesicles as part of a study on placental indoleamine 2,3-dioxygenase, the L-tryptophan-catabolising enzyme recently shown to regulate feto-maternal immunology. Brush border vesicle uptake of L-tryptophan is substantially into an osmotically active space. It is sodium independent and N-ethylmaleimide sensitive. Uptake of L-tryptophan, which is markedly stereospecific, has a Km of 26.3 microM and Vmax of 1.72 pmol (mg protein)(-1) s(-1) and is completely abolished by the L-system-specific substrate 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH). These findings are in keeping with L-tryptophan transport being exclusively via system L (induced by the heterodimeric heavy chain of CD98 and system L-amino acid transporter-1 (LAT-1)). 1-Methyl-tryptophan (which is a known competitive inhibitor of indoleamine 2,3-dioxygenase) is a competitive inhibitor of L-tryptophan flux through this transport system (Ki = 113 microM). Basal membrane transport of L-tryptophan is more complex. Uptake is slower than at the brush border and although, as in the brush border, uptake is sodium independent, it is less sensitive to N-ethylmaleimide. There is clear evidence that two systems contribute to basal membrane transport since BCH is (in sodium-free media) only a partial inhibitor whereas L-histidine and L-cysteine are fully effective. The simplest explanation of these and other findings is that the basal membrane possesses two systems, one of which is similar to that induced by the heavy chain of CD98 and system L-amino acid transporter-2 (LAT-2). The other appears to be system y+L since in the presence of BCH inhibition by L-leucine but not by L-lysine is sodium dependent. These findings suggest the existence of non-identical carrier-mediated transport systems for L-tryptophan in brush border and basal membranes. This asymmetry may explain net transplacental transfer of this amino acid.

Amino Acids↗
PubMed2001

The role of L-tryptophan transport in L-tryptophan degradation by indoleamine 2,3-dioxygenase in human placental explants.

The physiological importance of L-tryptophan transport for placental indoleamine 2,3-dioxygenase-mediated degradation of L-tryptophan has been studied using human placental chorionic villous explants. L-Tryptophan influx into villous explants is supported exclusively by transport system L and is substantially inhibited by the L-system-specific substrate 2-aminobicyclo-(2,2,1)-heptane-2-carboxylic acid (BCH) and also by 1-methyl-tryptophan which is also an inhibitor of indoleamine 2,3-dioxygenase. L-Tryptophan influx is enhanced 2.3-fold following in vitro culture of the villous explant. Interferon-gamma, which increases villous explant indoleamine 2,3-dioxygenase expression, has no effect on L-tryptophan influx. In explants both BCH and 1-methyl-tryptophan inhibit indoleamine 2,3-dioxygenase-mediated L-tryptophan degradation. This also applies when L-tryptophan degradation has been stimulated by interferon-gamma. These findings show transport of L-tryptophan into the trophoblast to be a rate-limiting step for indoleamine 2,3-dioxygenase-mediated L-tryptophan degradation and therefore for the normal physiology of mammalian pregnancy.

Amino Acids, Cyclic↗
PubMed2001

Low-threshold potassium channels and a low-threshold calcium channel regulate Ca2+ spike firing in the dendrites of cerebellar Purkinje neurons: a modeling study.

Various types of voltage-gated ion channels are distributed along the dendrites of neurons in the central nervous system. We have recently shown experimentally that the dendrites of cerebellar Purkinje neurons contain low-threshold voltage-gated Ca(2+) channels and low-threshold voltage-gated K+ channels. Although we found that these channels are involved in regulating the onset of Ca(2+)-dependent action potentials in the dendrites, we were unable to identify which of the known types of low-threshold Ca2+ channels and K+ channels were responsible, since there was no reliable method of discriminating between them. Here, we have built a detailed compartmental model of a Purkinje neuron by incorporating two types of low-threshold Ca2+ channel (T-type and class-E, or R-type) and two types of low-threshold K+ channel (A-type and D-type), in addition to another eight voltage-gated channel types, using a compartmental model neuron simulator. The model reproduces the basic features of the depolarization-induced responses of Purkinje neurons, such as fast Na+ spikes in the soma, Ca2+ spikes in the dendrites, the slow onset of Ca2+ spikes, repetitive Ca2+ spikes in the presence of TTX, the marked shortening of Ca2+ spike onset in the presence of 4-aminopydridine, and the longer Ca2+ spike onset in the presence of Ni2+. Our model shows that the D-type K+ channel and the class-E Ca2+ channel regulate the onset of depolarization-induced Ca2+ spikes in Purkinje neurons. These channels might be involved in integrating synaptic inputs in Purkinje neurons.

Action Potentials↗
PubMed2001

Flow cytometric analysis of hemetopoietic progenitor cells in peripheral blood stem cell harvest from patients with CD34 positive acute leukemia.

We analyzed CD34 positive cells in peripheral blood stem cell harvest (PBSCH) using flow cytometry. PBSCH from CD34 positive acute myelogeous leukemia (AML-M2) patient contained 1.87% CD34 positive cells, of which 1.21% was represented by MRD.PBSCH from CD34 positive acute lymphoblast leukemia (ALL) patient contained 3.14% CD34 positive cells, of which 0.11% was accounted for by minimal residual disease (MRD). If PBSCH from CD34 positive acute leukemia patient is analyzed for CD34 monoclonal antibody alone, the presence of CD34 positive MRD may escape attention so that CD34 positive hematopoietic progenitor cells may be overestimated. To avoid this risk, it is necessary to analyze PBSCH using both CD34 monoclonal antibody and characteristic markers of leukemia cells that were found pre-treatment.

Antigens, CD34↗
PubMed2001

GABAergic control of synaptic summation in hippocampal CA1 pyramidal neurons.

The primary function of neurons is to integrate synaptic inputs and to transmit the results to other cells. It was shown previously that separate excitatory inputs to hippocampal pyramidal neurons are summated nonlinearly. In the hippocampus, responses of pyramidal neurons are influenced by GABAergic inputs in feed-forward or feedback manner, and also by oscillatory network activities. It is likely that these GABAergic inputs regulate the way synaptic inputs are summated. To examine the roles of GABAergic inputs on synaptic summation, we made whole-cell recordings from the cell bodies of CA1 pyramidal neurons in rat hippocampal slices while stimulating two independent input pathways with short interstimulus intervals, and examined the manner by which postsynaptic potentials were summated. We found that: 1) the summation of the perforant pathway and the Schaffer collateral pathway inputs was sublinear when the interval between two inputs was shorter than 30 ms, 2) the blockade of GABA(A) receptors partially suppressed the sublinearity, and 3) further blockade of GABA(B) receptors removed the sublinearity totally. We also found that 4) the summation was superlinear under the concomitant blockade of GABA(A) and GABA(B) receptors when the two inputs arrived with no delay. Thus our study demonstrates that GABAergic inputs are responsible for keeping the summation of two separate inputs on CA1 pyramidal neurons sublinear.

Animals↗
PubMed2001

Cytokine expression in rat molar gingival periodontal tissues after topical application of lipopolysaccharide.

It is well known that proinflammatory cytokines produced by host cells play an important role in periodontal tissue destruction. However, the localization of the cytokines in in vivo periodontal tissues during development of periodontal disease has not been determined. Immunohistochemical expression of proinflammatory cytokines including IL-1alpha, IL-1beta, and TNF-alpha was examined at 1 and 3 h, and 1, 2, 3, and 7 days after topical application of lipopolysaccharide (LPS; 5 mg/ml in physiological saline) from E. coli into the rat molar gingival sulcus. In the normal periodontal tissues, a small number of cytokine-positive epithelial cells were seen in the junctional epithelium (JE), oral sulcular and oral gingival epithelium, in addition to macrophages infiltrating in the subjunctional epithelial area and osteoblasts lining the alveolar bone surface. Epithelial remnants of Malassez existing throughout periodontal ligament were intensely positive for IL-1beta but negative for the other two cytokines. At 3 h after the LPS treatment, almost all cells in the JE were strongly positive for the cytokines examined. In addition, several cytokine-positive cells, including neutrophils, macrophages, and fibroblasts, were seen in the subjunctional epithelial connective tissue. At day 2, expression of the cytokines in the JE gradually decreased, while cytokine-positive cells in the connective tissue increased in number. Positive staining of the cytokines was seen in osteoclasts and preosteoclasts which appeared along the alveolar bone margin in this period. The number of cytokine-positive cells decreased by day 7. These findings indicate that, in addition to macrophages, neutrophils, and fibroblasts, the JE cells are a potent source of TNF-alpha, IL-1alpha, and IL-1beta reacting to LPS application, and suggest that JE cells may play an important role in the first line of defense against LPS challenge, and the proinflammatory cytokines transiently produced by various host cells may be involved in the initiation of inflammation and subsequent periodontal tissue destruction.

Animals↗
PubMed2001

Erythrocyte creatine as a marker of excessive erythrocyte destruction due to hypersplenism in patients with liver cirrhosis.

OBJECTIVE: Erythrocyte creatine is a sensitive marker of erythrocyte age, and can be used to detect slight and continuous hemolysis. Excessive blood cell destruction caused by increased spleen function is important evidence of hypersplenism. This study evaluates the usefulness of erythrocyte creatine as a sensitive marker of excessive erythrocyte destruction due to hypersplenism in patients with liver cirrhosis. DESIGN AND METHOD: Erythrocyte creatine was determined by an enzymatic method in 50 patients with postnecrotic liver cirrhosis and 50 healthy controls. The spleen size was measured by ultrasonography and expressed as a spleen index. RESULTS: The patients with splenomegaly showed significantly higher erythrocyte creatine than those without splenomegaly (p < 0.005) and healthy controls (p < 0.001), but there was no significant difference in erythrocyte creatine between healthy controls and those without splenomegaly. Fourteen (93%) of the 15 patients with abnormally high erythrocyte creatine (> 1.8 micromol/g hemoglobin) had splenomegaly. There were no significant differences in reticulocyte count between healthy controls and the patients with and without splenomegaly. Erythrocyte creatine showed good correlations with spleen index (r = 0.67; p < 0.001) and reticulocytes (r = 0.63; p < 0.001). CONCLUSIONS: Erythrocyte creatine can be used for predicting erythropoietic status and estimating hypersplenism in patients with liver cirrhosis.

Aged↗
PubMed2001

Peripheral ameloblastoma: biological profile based on 160 cases from the literature.

The present profile of the peripheral ameloblastoma (PA) is based on a literature survey of 160 published tumour cases. The PA is an exophytic growth localized to the soft tissues overlying the tooth-bearing areas of the jaws, the initial diagnosis often being fibrous epulis. In most cases there is no radiological evidence of bone involvement, but a superficial bone erosion--known as cupping or saucerization--may be detected at operation. The PA accounts for 2-10% of all ameloblastomas. The overall average age is 52.1 years, slightly higher for males (52.9 years) than for females (50.6 years). Thus, the PA occurs at a significantly higher age than the intraosseous ameloblastoma (IA; 37.4 years). The male/female ratio amounts to 1.9:1, as opposed to 1.2:1 for the IA. The male/female ratio for the Japanese cases included in this survey is 2.5:1 as opposed to that of non-Japanese cases 1.4:1. As to the location of PA, the maxilla/mandible ratio is 1:2.6. The mandibular premolar region accounts for 32.6% of all sites. Five extra-gingival lesions have been reported under the term PA. As these cases most likely represent salivary gland tumours, they are not accepted under the diagnosis of PA. The odontogenic gingival epithelial hamartoma shows clinical, histological and behavioural features almost identical to the PA, and it is discussed whether this lesion and the PA should be considered one and the same entity. Pathogenetically, two major sources are discussed: remnants of the dental lamina and the oral surface epithelium. Histologically, the PA consists of proliferating odontogenic epithelium exhibiting the same histomorphological cell types and patterns as seen in the IA. The stroma is that of a mature, fibrous connective tissue. The indolent biological behaviour dictates a conservative therapeutical approach. It is discussed whether PA is a true neoplastic counterpart of the IA or rather an odontogenic hamartomatous lesion. Six cases of malignant PA have been reported.

Adult↗