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Biomedical subjects

O Saitoh

Publications and source records attributed to O Saitoh.

At least 73 records · Page 4Linked to original sources

Substrate specificity and distribution of UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase in the human stomach.

The detailed substrate specificity of the UDP-GalNAc:sialylparagloboside N-acetylgalactosaminyltransferase to form the Sd(a+) blood group active carbohydrate determinant GalNAc beta 1-4(NeuAc alpha 2-3)Gal was studied using a membrane fraction prepared from human gastric fundic mucosa. Various sialosylated oligosaccharides and gangliosides were examined as acceptor substrates. Oligosaccharide substrates were fluorescence-labelled with 2-aminopyridine, and the transferase activity was quantified by h.p.l.c. using a reversed-phase column. The structures of the products were determined by glycosidase degradation and proton n.m.r. 3'-Sialyl-lactose (II3NeuAcLac), 3'-sialyl-lactotetraose (IV3NeuAcLc4), and 3'-sialyl-lactoneotetraose (IV3NeuAcnLc4) were good substrates for the beta 1-4GalNAc transferase in gastric fundic mucosa, but 6'-sialyl-lactoneotetraose (IV6NeuAcnLc4) or 6'-sialyl-lactose (II6NeuAcLac) were not. Gangliosides with a terminal NeuAc alpha 2-3Gal residue such as GM3, sialylparagloboside, GM1b and GD1a were also studied. The activity of beta 1-4GalNAc transfer to sialylparagloboside was much higher than that to GM2, GM1b or GD1a in spite of them having the same terminal residue. Measurement of the activity of the beta 1-4GalNAc transferase in biopsy specimens demonstrated that the activity was localized in gastric fundic mucosa and was absent in pyloric mucosa, intestinal metaplasia and gastric cancer tissue. Thus the beta 1-4GalNAc transferase present specifically in fundic mucosa required a NeuAc alpha 2-3Gal residue connected to either type-1-chain or type-2-chain oligosaccharides. In glycolipids, the acceptor specificity was restricted to NeuAc alpha 2-3Gal beta 1-4GlcNAc because the NeuAc alpha 2-3Gal beta 1-3GalNAc structure in ganglio-series glycolipids was not a good acceptor substrate.

Aminopyridines↗

Differential glycosylation and cell surface expression of lysosomal membrane glycoproteins in sublines of a human colon cancer exhibiting distinct metastatic potentials.

Changes in the glycosylation of asparagine-linked oligosaccharides have been shown in various tumor cells, including human colon cancer. Attempts were made to elucidate the difference in Asn-linked oligo-saccharides attached to lysosomal membrane glycoproteins isolated from sublines of human colon carcinoma exhibiting high and low metastatic potentials in nude mice. Lysosomal membrane glycoproteins (lamp) 1 and 2 were immunoprecipitated from the cells after labeling with radioactive sugars, and the glycopeptides prepared were fractionated by serial lectin affinity chromatography employing immobilized concanavalin A, Datura stramonium agglutinin, and tomato lectin. Comparison of Asn-linked oligosaccharides from the different colonic carcinoma cells revealed the following features. First, the highly metastatic carcinoma cells express more poly-N-acetyllactosaminyl side chains with branched galactose residues than cells with low metastatic potential. Second, sialylation is more significant in the highly metastatic carcinoma cells than in the poorly metastatic ones. Conversely, N-acetyllactosamine units are less fucosylated in the highly metastatic cells than in poorly metastatic cells. These structural changes were apparently caused by the increase in sialyltransferase and the decrease in alpha 1----3 fucosyltransferase in the highly metastatic cells. The results also suggest that highly metastatic carcinoma cells express more sialyl Lex structures at the termini of poly-N-acetyllactosaminyl side chains than poorly metastatic carcinoma cells. Further, highly metastatic cells were found to express more lamp-1 and lamp-2 on the cell surface. These results were found to be correlated to the increased expression of sialyl Lex structures with high affinity binding of anti-sialyl Lex antibody on highly metastatic cells. Increased expression of sialyl Lex in the poly-N-acetyllactosamines of the cell surface may contribute to the metastatic behavior of the cells, assuming that this structure can serve as a better ligand for selectins present on endothelial cells and platelets.

Antibodies↗

Distributions of the Nd and P300 in a normal sample.

To obtain objective criteria for assessing the attentional and cognitive functioning of psychiatric populations, we attempted to standardize values of two components in Event-Related Potentials (ERPs), namely the attention-related negative potential (Nd) and the P300, in normal populations. The study consisted of 100 healthy volunteers (50 females, 50 males) who were given the task of making dichotic syllable discriminations requiring key-press responses. Their ages ranged between 18 and 59 years (mean +/- S.D., 32.3 +/- 11.3 years). Nd was found to be maximum in the Fz region, P300 being maximum in the Pz region. The means and standard deviations of Nd and P300 areas in their maximum regions were 554.1 +/- 307.8 microV ms and 2148.5 +/- 1248.5 microV ms, respectively. The transformation plot for symmetry indicated the suitable power of transformation to be 1/2 for both Nd and P300 distributions. After being transformed into square-root values, the distribution patterns of Nd and P300 areas were examined. When the lower limit of normal values was tentatively assigned to mean -2 S.D. using square-root transformed data for both Nd and P300, 97% of the subjects were found to display values above the lower normal limit for Nd, and 98% for the P300. Neither, Nd nor P300 areas correlated with age, while P300 latencies displayed a weak positive correlation with age. Females displayed relatively larger values than males for Nd and P300 areas and P300-peak amplitudes. However, the differences between females and males were not statistically significant. Females and males showed nearly equal P300-peak latencies.

Adolescent↗

cis-4-Hydroxy-L-proline and ethyl-3,4-dihydroxybenzoate prevent myogenesis of C2C12 muscle cells and block MyoD1 and myogenin expression.

cis-4-Hydroxy-L-proline (cis-OH-Pro) and ethyl-3,4-dihydroxybenzoate (EDHB), two distinct inhibitors of collagen synthesis, prevented myogenesis in C2C12 mouse skeletal muscle cells. Both inhibitors blocked myotube formation and the expression of sarcomeric myosin heavy chain. Northern blot analysis showed that cis-OH-Pro- and EDHB-treated C2C12 muscle cells did not express the myogenic regulatory genes, MyoD1 and myogenin, but continued to express non-muscle isoforms of actin (beta and gamma) and alpha-tropomyosin. 10TFL2-3B cells, a C3H10T1/2 cell line permanently transfected with myogenin cDNA, constitutively expressed exogenous myogenin in the presence of cis-OH-Pro but failed to activate endogenous myogenin and to undergo myogenesis. These results demonstrate that commitment to terminal differentiation and activation of myogenic regulatory genes requires active synthesis of the extracellular matrix component collagen.

Animals↗

Information dysregulation and event-related potentials in schizophrenia.

Clinical experiences indicate that schizophrenic cognitive disturbances may be partly due to the unsuccessful construction of their cognitive context. In this article, two experiments are introduced in which schizophrenic deficits in utilization of information for construction of effective cognitive context were examined through measurements of event-related potentials, particularly P300s. In a 3-tone discrimination task, schizophrenic subjects failed to elicit P300s to frequent nontarget tones that were as important as standard tones for performing the required task. In the second task, consisting of detecting two consecutive identical tones, unlike healthy controls, schizophrenic subjects failed to display P300s to nontarget tones at target position. These results are discussed in relation to ineffective context construction of schizophrenic subjects. Moreover, a single-trial analysis of relationships between P300s and reaction times in the 3-tone discrimination task disclosed a loose coupling between the stimulus set and response set in patients.

Adult↗

Histological study on bone response to resin-hydroxyapatite.

Resin-Hydroxyapatite has been developed to take the advantages of bioactivity of hydroxyapatite (HAP) and plasticity of resin (Bis-GMA). It was composed of 75% HAP, 15% Bis-GMA and 10% quartz reinforced filler. Tested pellets of Resin-HAP as well as its constituents were implanted into subcutaneous connective tissue of femurs of adult dogs. Histological observation at 12 weeks postoperatively showed that each pellet was surrounded by the fibrous connective tissue almost free of the inflammatory cells. The thickness of the fibrous layer around HAP was relatively thin, while those around Bis-GMA and Resin-HAP showed some variance. Taking our previous data into consideration, it was suggested that Resin-HAP had bio-compatibility both in soft tissue and bone, although its long-term stability as well as its stability under function should further be examined.

Animals↗

Expression of aberrant O-glycans attached to leukosialin in differentiation-deficient HL-60 cells.

Promyelocytic leukemia HL-60 cells can be induced to differentiate into granulocytic cells by various agents including retinoic acid (RA), dimethyl sulfoxide, and 6-thioguanine (6-TG). Although the induced cells are no longer capable of proliferation, a few cells continue to divide in the presence of inducers, and these cells are resistant to terminal differentiation by these inducers (R. E. Gallagher, D. A. Giangiulio, C-S. Chang, C. J. Glover, and R. L. Felsted, Blood, 68: 1402-1406, 1986). The present study examined the structures of O-glycans attached to leukosialin, a major sialoglycoprotein in HL-60 cells, and the activities of glycosyltransferases involved in O-glycan synthesis. Leukosialin from RA-resistant and 6-TG-resistant HL-60 sublines migrated much more slowly than those from wild-type HL-60 cells when applied to sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The dimethyl sulfoxide-resistant HL-60 subline, on the other hand, expressed leukosialin with a molecular weight similar to wild-type HL-60 cells. RA-resistant and 6-TG-resistant HL-60 cells were found to express a significant amount of tetrasaccharides that contain no sialic acid residue, while wild-type HL-60 cells expressed mainly disialosyl hexasaccharides and contained no detectable amount of asialo-oligosaccharides. Furthermore, wild-type HL-60 cells treated with the inducers for 4 days were found to express the same saccharides present in untreated wild-type HL-60 cells, indicating that the altered O-glycans present in RA and 6-TG sublines were not caused by a direct effect of these agents but rather are intrinsically unique to these sublines. To better understand the mechanisms underlying the differences in O-glycans, the activities of four sialyltransferases were measured: Gal beta 1----3GalNAc alpha 2----3sialyltransferase, Gal beta 1----4(3) GlcNAc alpha 2----3sialyltransferase, Gal beta 1----4GlcNAc alpha 2----6sialyltransferase, and GalNAc alpha 2----6sialyltransferase. Among them, Gal beta 1----3GalNAc alpha 2----3sialyltransferase and Gal beta 1----4(3)GlcNAc alpha 2----3sialyltransferase were much lower in the RA- or 6-TG-resistant HL-60 subline than in wild-type HL-60 cells. These findings indicate that the differences in O-glycans are due to the differences in alpha 2----3sialyltransferase activities. These results strongly suggest that O-glycans associated with leukosialin may play some role in HL-60 cell differentiation.

Antigens, CD↗

T-lymphocytic leukemia expresses complex, branched O-linked oligosaccharides on a major sialoglycoprotein, leukosialin.

Leukocytes express a major sialoglycoprotein, leukosialin, of which the apparent molecular weight (mol wt) can be variable according to the differences in O-glycans attached to this molecule. In the present study, we analyzed the structures of O-glycans attached to leukosialin present in various T-lymphocytic leukemia cells. T-lymphoid cells from patients with acute T-lymphocytic leukemia express a large amount of the branched hexasaccharides, NeuNAc alpha 2----3Gal beta 1----3(NeuNAc alpha 2----3Gal beta 1----4GlcNAc beta 1----6)GalNAc, which are also expressed in activated normal T lymphocytes, but that are almost absent in resting normal T lymphocytes. T-lymphoid cells from patients with chronic T-lymphocytic leukemia, on the other hand, mainly express the tetrasaccharides NeuNAc alpha 2----3Gal beta 1----3(NeuNAc alpha 2----6)GalNAc on leukosialin, but they also express a small significant amount of the hexasaccharides. The same hexasaccharides can be detected in thymocytes. The increased amount of the hexasaccharides in acute leukemia is associated with increased activity of beta 1----6 GlcNAc-transferase, a key enzyme in forming the hexasaccharides. Immunoblot analysis of cell lysates showed that monoclonal antibody (MoAb) T-305 reacts preferentially with leukosialin of high mol wt containing the hexasaccharides. These findings indicate that T-lymphocytic leukemia cells reexpress the oligosaccharides present in immature cells.

Antibodies, Monoclonal↗

Muscle-specific RNA processing continues in the absence of myogenin expression.

During muscle development, muscle-specific gene expression is achieved by a complex set of transcriptional and post-transcriptional mechanisms. We examined the requirement of new protein synthesis and myogenin expression for muscle-specific tropomyosin expression during myogenesis of the rat muscle cell line L6E9. The results show that new protein synthesis is required for both muscle gene transcription and muscle-specific alternate RNA processing. However, once initiated, the synthesis of muscle tropomyosin mRNA continued in the presence of cycloheximide, while the expression of muscle actin, myosin heavy chain, and myogenin mRNA was abolished. These results suggest that muscle-specific processing of tropomyosin transcripts can continue to occur in the absence of myogenin expression unlike the expression of muscle actin and myosin heavy chain mRNAs. However, the transfection of myogenin cDNA into nonmuscle cells induced muscle-specific RNA processing of tropomyosin, suggesting indirect involvement of myogenic factors in the initiation of muscle-specific RNA processing pathways.

Actins↗

Psychiatric patients showing irregular beta activities in EEGs and treatment with antiepileptic drugs: a report of 15 cases.

Fifteen psychiatric cases are reported who were clinically diagnosed as schizophrenic, affective disorders, or neurotic, but resisted standard medication regimens, all showing irregular beta activities on EEGs. The cases tended to display symptoms in common, such as dysphoria, emotional instability or frequent physical complaints. These characteristic symptoms share something mutually with the symptoms shown in some epileptic patients or psychiatric patients with epileptic EEG abnormalities without clinical seizures. Antiepileptic drugs seemed more specifically effective to the above symptoms. More than half of these cases showed improvement on EEG findings such as a decrease in irregular beta activities and an increase in rhythmicity or regularity of alpha activities along with clinical improvement with the administration of adjunctive antiepileptic drugs. These results suggest that the adjunctive administration of antiepileptic drugs to patients with irregular beta activities on EEGs is clinically useful and an EEG examination has much value in psychiatric practice to find the criteria of drug therapy.

Adult↗

Psychological intervention can partly alter P300-amplitude abnormalities in schizophrenics.

In the present study, we investigated whether psychological interventions can alter P300-abnormalities, specifically enhancing reduced P300-amplitudes, in schizophrenics. A three-tone discrimination task was employed for recording P300s, in which psychological intervention to facilitate target detection was performed through delivering a buzzer sound one second after each designated target tone that informed its occurrence. This procedure was done exclusively during the third and fourth sessions among the six sessions in total. When the data for all the patients were analyzed as a whole, no significant change was observed. However, when the patients were broken down into two groups based on the P300-amplitudes in the first and second sessions, significant effects of the intervention emerged. The group with smaller P300-amplitudes showed a significant increase in P300-amplitudes as well as improved performance levels during and after the intervention sessions. On the contrary, the group with larger P300-amplitudes displayed a significant decrease in P300-amplitudes in these sessions. Interestingly, the above results were consistent with the subjective difficulty changes experienced by the patients through the sessions. Overall, the above results indicate that psychological interventions can partly enhance reduced P300-amplitudes in schizophrenics.

Adult↗

A study on gastric ulcers induced by long-term fasting in rats.

The present study was performed to investigate the roles of intragastric pH and PAS positive mucus as to the incidence of gastric ulcers after long-term fasting in rats. Gastric ulcers in the forestomach and corpus were found at 4 and 6 days after fasting, respectively. The intragastric pH was significantly reduced at 1, 2, 3, 4, 6 and 7 days after fasting, but tended to have returned to control levels at 8 days. PAS positive mucus was significantly increased at 2 to 7 days, but there was no significant change at 8 days. From these results, it seems that the aggressive factor is not involved, but attenuation of the defensive factor is important as to the incidence of this type of ulcer. As one of the causes of this attenuation, a decrease in the gastric mucus volume was suggested.

Animals↗

Distribution of microtubules and other cytoskeletal filaments during myotube elongation as revealed by fluorescence microscopy.

Distribution of microtubules and other cytoskeletal filaments in growing skeletal muscle cells (myotubes) was studied in vitro by fluorescence microscopy using fluorescin-labeled antibodies and phalloidin, a specific antiactin drug. In the distal elongating tips of myotubes, microtubules were the major cytoskeletal elements; actin and intermediate filaments were much less abundant. On the other hand, colcemid- and nocodozole-treatments caused disruption of microtubles and also prompt retraction of growth tips to form myosacs, a type of deformed myotube. Actin filaments remained unaffected during the retraction. The difference in the distribution of the 3 cytoskeletal filaments in the region of growth tips was most remarkable in the case of those myotubes in the process of recovery from myosacs. In an early phase of recovery, the cellular processes extending from myosacs were enriched with both microtubules and intermediate filaments, but not with actin filaments. Later, when the processes became further developed, intermediate filaments were scarce at the extreme ends. Fluorescein-labeled actin introduced by a micro-injection method was minimally incorporated into filaments in the cellular processes. We conclude that microtubules make up the cytoskeletal element which is most responsible for elongation or spreading of growth tips of myotubes in vitro.

Actins↗

Airway hyperresponsiveness after endotoxin inhalation depends on leukocyte infiltration.

We have investigated the relationship between acute airway hyperresponsiveness and polymorphonuclear cell (PMN) migration in airways following saline or endotoxin (ET) aerosol inhalation in 63 guinea pigs. In 20 of these animals, granulocytopenia was induced by prior treatment with hydroxyurea (HU). Airway responsiveness (AR) to inhaled methacholine, together with leukocyte counts in bronchoalveolar lavage fluid (BALF) and in tracheal epithelium, was also examined before and at 30 min, 1, 3 and 6 hr after saline or ET inhalation. In saline inhalation groups, AR and PMN counts in BALF and in tracheal epithelium did not differ from control animals at any time points. However, in ET inhalation groups AR increased significantly at 1 and 3 hr and returned to the pre-exposure level at 6 hr. This period of hyperresponsiveness was associated with an increment of PMN migration into tracheal epithelium. However, the increment of PMN counts in BALF was delayed to 3 and 6 hr after ET. In HU treated animals, AR and PMN counts in BALF and in tracheal epithelium did not change during 6 hr after ET inhalation. These results suggest that the hyperresponsiveness induced by ET inhalation either depends upon PMN migration into the tracheal epithelium.

Administration, Inhalation↗