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Biomedical subjects

T Saitoh

Publications and source records attributed to T Saitoh.

At least 37 records · Page 2Linked to original sources
PubMed2001

Somatic mutations and single nucleotide polymorphisms of base excision repair genes involved in the repair of 8-hydroxyguanine in damaged DNA.

To elucidate the involvement of 8-hydroxyguanine (oh(8)G) repair genes in human lung carcinogenesis, 47 lung cancer cell lines and 55 primary lung cancers were examined for somatic mutations and genetic polymorphisms in all coding exons of the MYH and APEX genes, and exon 8 of the OGG1 gene by polymerase chain reaction-single strand conformation polymorphism analysis. In the MYH gene, one missense mutation was detected in a cell line, NCI-H157, whereas no mutations were detected in primary cancers. There were no mutations in the APEX and OGG1 genes in the cell lines or primary cancers. Ten single nucleotide polymorphisms (SNPs) were identified, and seven of them were accompanied by amino acid substitutions. Differences in the oh(8)G repair activities of MYH, APEX and OGG1 proteins due to somatic mutations and SNPs can be involved in human carcinogenesis.

Carbon-Oxygen Lyases↗
PubMed2001

Screening of a unique lectin from 16 cultivable mushrooms with hybrid glycoprotein and neoproteoglycan probes and purification of a novel N-acetylglucosamine-specific lectin from Oudemansiella platyphylla fruiting body.

Hybrid glycoprotein and neoproteoglycan probes were prepared by coupling various glycoproteins or polysaccharides to peroxidase or biotinyl bovine serum albumin, respectively. Lectins recognizable by the neoglycoconjugate probes were extracted from 16 cultivable mushrooms. Dot-blot assay revealed five extracts to be reactive with only hybrid glycoprotein probes, but others also reacted with neoproteoglycan probes. According to the reactivity pattern with probe screening, the one lectin from Oudemansiella platyphylla extract (OPL) bound best with asialotransferrin-- and asialoagalactotransferrin--peroxidase probes and was isolated using an asialotransferrin column, but it did not bind with other hybrid glycoprotein or neoproteoglycan probes. OPL, consisting of two polypeptides with high homology in the N-terminal amino acid sequences, exhibited weak hemagglutinating activity. Purified OPL specifically bound the beta-GlcNAc probe among various biotinylated polymeric sugar probes, while it exhibited essentially the same binding specificity toward neoglycoconjugate probes as that of the crude extract, showing a preference for the asialobiantennary complex type of N-linked glycans. These results indicate that the neoglycoconjugate probes are valuable in lectin screening.

Acetylglucosamine↗
PubMed2001

Molecular cloning and characterization of FRAT2, encoding a positive regulator of the WNT signaling pathway.

FRAT1 positively regulates the WNT signaling pathway by stabilizing beta-catenin through the association with glycogen synthase kinase-3beta. Here, we have cloned FRAT2 cDNAs, spanning the complete coding sequence, from a human fetal lung cDNA library. FRAT2 encoded 233 amino-acid protein, which showed 77.3% total amino-acid identity with FRAT1. FRAT2 and FRAT1 were more homologous in the acidic domain (96% identity), the proline-rich domain (92% identity), and the GSK-3beta binding domain (100% identity). The FRAT2 gene was mapped to human chromosome 10q24.1. The FRAT2 mRNA of 2.4-kb in size was relatively highly expressed in MKN45 (gastric cancer), HeLa S3 (cervical cancer), and K-562 (chronic myelogenous leukemia). Xenopus axis duplication assay revealed that the wild-type FRAT2 mRNA, but not the mutant FRAT2 mRNA lacking the acidic domain and the proline-rich domain, has the capacity to induce the secondary axis. These results indicate that FRAT2, just like FRAT1, functions as a positive regulator of the WNT signaling pathway. Thus, up-regulation of FRAT2 in human cancer might be implicated in carcinogenesis through activation of the WNT signaling pathway.

Adaptor Proteins, Signal Transducing↗
PubMed2001

Adrenomedullin receptors in rat choroid plexus.

To characterize transmembrane signaling of adrenomedullin (AM) in the choroid plexus, we studied the effects of AM on cyclic AMP (cAMP) and cyclic GMP (cGMP) levels as well as expression of mRNA for AM receptor in the rat choroid plexus slices. AM or calcitonin gene-related peptide (CGRP) increased cAMP (but not cGMP) level in a concentration-dependent manner, with AM being much more potently than CGRP. AM mRNA as well as calcitonin-receptor-like receptor mRNA and receptor-activity-modifying protein 2 mRNA, were highly expressed in the choroid plexus. Our biochemical and pharmacological studies may raise the possibility that choroid plexus secretes AM into the cerebrospinal fluid, and AM regulates choroid plexus function in an autocrine/paracrine manner via acting on AM-specific receptors.

Adrenomedullin↗
PubMed2001

Risk factors for early death in patients undergoing treatment for multiple myeloma.

The survival time of myeloma patients improved from a few months to many years after treatment with melphalan. Perhaps chemotherapy more intensive than melphalan-prednisolone should be administered to patients at risk of early death. Therefore, early death must be accurately predicted. We analyzed 93 patients with recently diagnosed myeloma and found that 13 (14%) died within 6 months (early death). The most common cause of death was bacterial and fungal pneumonia when myeloma became uncontrollable. The response to conventional chemotherapy was poorer in patients at high risk of early death than the control group. Multivariate analysis showed that the serum level of beta-2 microglobulin was the only value that predicted early death.

Aged↗
PubMed2001

Effects of single and repeated administration of 1,2,3,4-tetrahydroisoquinoline analogs on the binding of [11C]raclopride to dopamine D2 receptors in the mouse brain.

We investigated the effects of intraperitoneal injection of 1,2,3,4-tetrahydroisoquinoline (TIQ) analogs and 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) on the binding of [11C]raclopride to striatal dopamine D2 receptors in mice. The binding of [11C]raclopride, but not of [11C]N-methylspiperone or [11C]nemonapride with higher affinity, to the receptors was significantly decreased immediately after TIQ injection. Neither a dopamine transporter blocker induced such effect nor TIQ affected the dopamine transporter-radioligand binding. Among the compounds investigated, including parkinsonism-inducing TIQ and (R/S)-1-benzyl-TIQ, parkinsonism-preventing (R)- and (S)-1-methyl-TIQ, and probable N-methylated metabolites of TIQ and 1-methyl-TIQ, TIQ and (S)-1-methyl-TIQ had the strongest effect on the binding of [11C]raclopride, and N-methylated derivatives showed less of an effect than the respective parent compounds. The decrease in the binding of [11C]raclopride continued for 7 hours and was followed by an increase until 10 days after the single and subchronic administration of TIQ. These findings suggest that TIQ analogs profoundly stimulated dopamine release which resulted in the competitive inhibition of the binding of [11C]raclopride to dopamine D2 receptors, but did not induce degeneration of the receptors.

Animals↗
PubMed2001

Selective expression of aquaporin 1, 4 and 5 in the rat middle ear.

The middle ear cavity is an air-filled space that must be maintained for effective sound transmission to the inner ear. To examine the mechanisms of water homeostasis in the middle ear, we investigated whether aquaporins (AQPs), a family of water-permeable channels, were expressed in the middle ear. Reverse transcription-polymerase chain reaction and immunoblot analyses revealed that mRNAs encoding AQP1, 4 and 5 (but not 2 or 3) subtypes were expressed in rat middle ear epithelium; AQP1, 4 and 5 were detected as 28-, 30- and 30-kDa proteins, respectively. Immunohistochemical analysis showed that AQP1 was localized at capillary endothelial cells and fibroblasts in lamina propria mucosae; AQP4 was present solely at the basolateral membrane of ciliated cells, whereas AQP5 was on the apical surface of ciliated cells as well as of flat and columnar epithelial cells. The characteristic different localizations of AQP1, 4 and 5 subtypes in the middle ear suggest that middle ear water homeostasis requires the coordinated operation of these AQPs.

Animals↗
PubMed2001

Intragastric acidity and circadian rhythm.

Most patients with peptic ulcers or gastroesophageal reflex disease develop subjective symptoms of epigastralgia and retrosternal pain during the period of time from the middle of the night to the early dawn (nocturnal pain). Such pain often disappears before breakfast. Disturbed circadian rhythm of gastric acid secretion may have a close relationship with the onset and aggravation of acid-related diseases. On the other hand, Helicobacter pylori has been considered to be an etiological agent of duodenal ulcer, and H. pylori eradication has been conducted in patients with gastritis and peptic ulcers. However, such eradication therapy sometimes results in the onset or deterioration of gastroesophageal reflux diseases. In this context, the question of whether the circadian rhythm of gastric acid secretion varies in accordance with the presence or absence of H. pylori infection is of interest. In the present study, we examined the fluctuation in intragastric acidity via a portable pH meter in 10 H. pylori-positive and 10 H. pylori-negative subjects. As a result, a significant difference in the circadian rhythmicity was observed between the H. pylori-negative and the H. pylori-positive group, with mean values for each parameter of 28.1 and 13.3 for amplitude, 22.7 and 12.4 for the midline-estimating statistic of rhythm (MESOR), and 324.0 and 321.0 for acrophase, respectively (P < 0.001). In both H. pylori-positive and negative groups, a tendency was observed toward an increase in intragastric acidity during the time period from the middle of the night to the early dawn, and toward a decrease in intragastric acidity during the early morning. In the H. pylori-positive group, the values for intragastric acidity over time were lower, and the degree of amplitude was smaller as compared to the H. pylori-negative group. Further, H. pylori-positive individuals were at a more advanced stage of the disease.

Adult↗
PubMed2001

Synthesis and chain length-anti-HIV activity relationship of fully N- and O-sulfated homooligomers of tyrosine.

Fully N- and O-sulfated homooligomers from octamer to nonadecamer of tyrosine were obtained as their sodium salts, aO3S-[Tyr(SO3Na)]n-ONa (n = 8-19), from reaction mixtures of tyrosine with sulfur trioxide trimethylamine and pyridine comlexes, respectively, in pyridine. Their anti-HIV activity increased along with the increase of the chain length up to the dodecamer, maintained the same level to the length of the heptadecamer and then decreased. The maximal activity level was the same as or higher than that of dextran and curdlan sulfates.

Anti-HIV Agents↗
PubMed2001

Synthesis and anti-HIV activity of nonatyrosine N- and O1-9-decasulfate.

To develop a potent and effective anti-HIV compound with a definite polyanionic structure, synthesis of oligotyrosine sulfates by oligomerization with simultaneous sulfation of tyrosine was tried. One component was successfully isolated from the mixture containing many products as its sodium salt (Y-ART-4) and was identified as the salt of nonatyrosine N- and O1-9-decasulfate, NaO3S-[Tyr(SO3Na)]9-ONa. Anti-HIV activity of Y-ART-4, determined from the protection it provided against HIV-induced cytopathic effects, was almost the same with that of dextran sulfate and curdlan sulfate.

Anti-HIV Agents↗
PubMed2001

G649, an allelic variant of the human H2 receptor with low basal activity, is resistant to upregulation upon antagonist exposure.

Orange et al reported an allelic variant of the human histamine H2 receptor, in which adenine 649 was replaced with guanine, to be more frequent in the schizophrenic population than controls in British Caucasians. The A649 to G change causes an Asn to Asp transition at amino acid position 217 in the third intracellular region, which is postulated to be important for receptor function. Herein, we analyzed the functional significance of this variant using wild-type and variant receptors expressed in Chinese hamster ovary cells. The variant receptor was associated with markedly lower basal cAMP productions than the wild-type receptor. Histamine-dependent cAMP productions via the variant receptor were lower as well. Treatment of cells expressing variant receptors with 10(-5) M ranitidine for 24 h resulted in a reduced degree of receptor upregulation as compared with the wild-type receptor. Thus, this is the first report of an allelic variant of the human H2 receptor which confers altered receptor function. To analyze gastric acid secretion in individuals with this variant, we examined 100 Japanese control subjects. However, neither heterozygotes nor homozygotes were found, suggesting that this variant, if present, is uncommon in the Japanese population.

Alleles↗
PubMed2001

Effect of cytokines on growth and differentiation of leukaemic cells with translocation t(6;9)(p23;q34).

The translocation t(6;9)(p23;q34) is detected infrequently in subtypes of haematological malignancies including acute myelogenous leukaemia (AML) and myelodysplastic syndrome (MDS). Although the t(6;9) leukaemia is commonly associated with bone marrow basophilia, the cytological characteristics of leukaemic cells are unclear. In the current study, we examined the in vitro effects of several cytokines on growth and differentiation of t(6;9) leukaemic cells. Isolated bone marrow mononuclear cells from four patients with t(6;9) (two MDS and two AML) were cultured for 14 d in the presence or absence of each cytokine. At the end of culture, viable cells were counted, and their histology was examined. Bone marrow cells obtained from 22 patients (10 AML, six AML from MDS, six MDS) lacking t(6;9) were used as controls. Compared with control cultures, significantly higher numbers of blasts appeared in the culture of bone marrow cells from t(6;9)-positive patients in response to stimulation with granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage CSF (GM-CSF) or interleukin 3 (IL-3). Stem cell factor (SCF) had little effect. Neutrophil counts were also significantly increased in the presence of G-CSF or IL-3. SCF and IL-3 were potent in increasing basophil counts from t(6;9)-positive cultures. These findings suggest that bone marrow cells obtained from t(6;9) patients are highly sensitive to growth- and/or differentiation-promoting cytokines. Special attention should be paid to the use of "therapeutic" cytokines in these patients.

Adolescent↗
PubMed2001

In vitro cytochrome P450-mediated hepatic activities for five substrates in specific pathogen free chickens.

Total hepatic microsomal cytochrome P450 (CYP) content as well as in vitro CYP mediated activities for five substrates [bufuralol 1-hydroxylation, ethoxyresorufin O-deethylation, S-mephenytoin 4-hydroxylation, testosterone 6beta-hydroxylation, and tolbutamide hydroxylation] were measured in specific pathogen free male Japanese leghorn chickens and male beagle dogs. The Vmax, Km and intrinsic clearance (Vmax/Km) for these substrates were calculated and compared between animal species in order to evaluate the drug catalytic activity in chicken liver. The total CYP content in chicken (0.296 +/- 0.04 nmol/mg microsomal protein) was close to levels reported for other species including humans, cats, pigs and some nonmammalian vertebrates (e.g. snakes, frogs and trout fish), but was lower than levels measured in dogs (1.11 +/- 0.22) or recorded in guinea-pigs, hamsters, monkeys, mice, rabbits, rats, horse and ruminants. Bufuralol 1-hydroxylation, ethoxyresorufin O-deethylation, S-mephenytoin 4-hydroxylation, and testosterone 6beta-hydroxylation were lower in chickens than in dogs based on intrinsic clearance. On the other hand, tolbutamide hydroxylation was markedly higher in chickens than in dogs.

Animals↗
PubMed2001

Rapid induction and Ca(2+) influx-mediated suppression of vitamin D3 up-regulated protein 1 (VDUP1) mRNA in cerebellar granule neurons undergoing apoptosis.

Cerebellar granule neurons (CGNs) grown under depolarizing conditions with high K(+) (HK; 30 mM) undergo apoptosis following replacement of HK by physiological K(+) (5.4 mM). Differential display analysis identified eight genes up-regulated in this paradigm of apoptosis. Vitamin D3 up-regulated protein 1 (VDUP1) mRNA was markedly up-regulated as early as 2 h following HK withdrawal. VDUP1 mRNA was up-regulated in other paradigms of neuronal apoptosis as well both in vitro and in vivo. HK effectively suppressed the up-regulation of VDUP1 mRNA in CGNs undergoing apoptosis via Ca(2+) influx through voltage-dependent L-type Ca(2+) channels, which did not require de novo protein synthesis. The up-regulation occurred in parallel with that of the c-jun transcript and c-jun protein phosphorylation. Moreover, SB203580, p38 mitogen-activated protein kinase inhibitor, suppressed up-regulation of both c-jun and VDUP1 mRNAs, and c-jun phosphorylation in CGNs undergoing apoptosis. IGF-1, one of the neuroprotective agents for CGNs, also inhibited VDUP1 mRNA up-regulation through a phosphoinositide 3 kinase-dependent pathway. These results suggest that the VDUP1 gene is a novel member of early response genes in neuronal apoptosis whose expression is directly regulated by Ca(2+) influx and coordinately regulated with the transcription factor c-jun in CGNs.

Animals↗
PubMed2001

Evaluation of neurotoxicity of TIQ and MPTP and of parkinsonism-preventing effect of 1-MeTIQ by in vivo measurement of pre-synaptic dopamine transporters and post-synaptic dopamine D(2) receptors in the mouse striatum.

Parkinsonism-inducing neurotoxicity of 1,2,3,4-tetrahydroisoquinoline (TIQ), as contrasted to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and parkinsonism-preventing effect of 1-methyl-1,2,3,4-tetrahydroisoquinoline (1-MeTIQ) have been investigated in mice by measuring their effects on the in vivo binding of radioligand to pre-synaptic dopamine transporters (DATs) or to dopamine D(2) receptors (D2R) in the striatum. A significant reduction of the ligand-DATs binding was found in the mice treated with MPTP, but not with TIQ, under the dosage inducing behavioral abnormality and loss of tyrosine hydroxylase-positive cells in the substantia nigra. A slight decrease in the ligand-DATs binding was observed in the mice given a larger dose of TIQ. Compensatory up-regulation in the post-synaptic D2Rs was found in the MPTP-treated mice. Pre-treatment with (S)-enantiomer, but not (R)-enantiomer, of 1-MeTIQ prevented the degeneration of DATs to some extent. We concluded that the TIQ-induced parkinsonism model is different from the MPTP-induced model as evaluated by the radioligand-DATs binding and that (S)-1-MeTIQ has a preventing effect for the degeneration of the DATs to a certain extent.

1-Methyl-4-phenyl-1,2,3,6-tetrahydropyridine↗
PubMed2001

OGG1 protein suppresses G:C-->T:A mutation in a shuttle vector containing 8-hydroxyguanine in human cells.

8-Hydroxyguanine (8-OHG) is an oxidatively damaged mutagenic base which causes G:C-->T:A transversions in DNA. OGG1 was cloned as a human gene encoding a DNA glycosylase that specifically excises 8-OHG from DNA in vitro. However, it was not clear whether OGG1 protein suppresses G:C-->T:A transversions caused by 8-OHG in human cells in vivo. In the present study we have examined the ability of OGG1 protein to suppress G:C-->T:A transversions caused by 8-OHG in human cells by bacterial suppressor tRNA (supF) forward mutation assay using a shuttle vector DNA, pMY189. Introduction of a single 8-OHG residue at position 159 of the supF gene in plasmid pMY189 resulted in a 130-fold increase in mutation frequency compared with untreated plasmid pMY189 after replication in the NCI-H1299 human lung cancer cell line. G:C-->T:A transversions at position 159 were detected in >90% of the supF mutants from the 8-OHG-containing plasmid. The mutation frequency of the 8-OHG-containing plasmid was significantly reduced by overexpression of OGG1 protein in NCI-H1299 cells and, in particular, the occurrence of G:C-->T:A transversion at position 159 in the supF gene was suppressed. Furthermore, frequencies and spectra of mutations of the untreated pMY189 plasmid did not differ significantly with overexpression of OGG1 protein. These results indicate that OGG1 protein has the ability to suppress G:C-->T:A transversions caused by 8-OHG in human cells in vivo.

Base Sequence↗
PubMed2001

Effects of endurance training on resting and post-exercise cardiac autonomic control.

PURPOSE: Endurance training induces reductions in both resting and postexercise heart rate (HR). If adaptation in cardiac autonomic regulation is a contributing factor in these reductions, changes in cardiac autonomic nervous system (ANS) should correspond to those in HR during an endurance-training program. We investigated the changes in resting and postexercise HR variabilities (both in the time and frequency domain) over a 6-wk training program. METHODS: HR variability was measured five times in an endurance-training group (N = 7) and four times in a control group (N = 5) during the course of study. RESULTS: Endurance training decreased HR and increased indices of parasympathetic modulation measured both at rest and during postexercise recovery periods. Noteworthy is that no changes in either HR or indices of ANS modulation measured during postexercise recovery periods were detectable after the first 7 d of the study despite continued changes in resting HR and indices in ANS modulation measured between the 7th and 42nd days of the endurance-training program. CONCLUSIONS: The study demonstrates that with endurance-training changes in cardiac ANS modulation partly contribute to a decrease in HR at rest and during postexercise recovery period, and that adaptation of the cardiac autonomic control occurs sooner in immediate postexercise periods than at rest.

Adaptation, Physiological↗