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S Ueda

Publications and source records attributed to S Ueda.

At least 163 records · Page 9Linked to original sources

The different mobility of complementary strands depends on the proportion AC/GT.

The electrophoretic mobility of DNA fragments on denaturing polyacrylamide gel depends on various factors. One of these is the base composition of a single-stranded DNA (ssDNA). We confirmed that one strand and its complementary strand of polymerase chain reaction (PCR) products migrated with different mobilities in all alleles detected at 12 out of the 13 short tandem repeat (STR) loci studied. The mobility differences between complementary strands (MD) were also observed regardless of end-polishing with Pfu DNA polymerase. MD was therefore not influenced by additional nucleotides to each strand of the PCR products. We then analyzed the relation between MD and the base composition using one representative allele at each of the 13 loci. The results indicated that MD was affected by the adenine plus cytosine (AC) content in the ssDNA and was proportional to the values of the AC content divided by the guanine plus thymine (GT) content in the AC-rich strand (the proportion AC/GT). When the proportion AC/GT was well-balanced, MD decreased. The same tendency was observed even in the end-polished strands. In this study, the electrophoretic mobility of an ssDNA on denaturing polyacrylamide gels was shown to depend on the proportion AC/GT. Unless the same side of the PCR products is labelled in the context of a PCR-based STR typing, distinct alleles may be mistaken for identical ones because of the different mobility of complementary strands. Accordingly, the labelled strand should be described if only one strand of the PCR products is detected. When using an allelic ladder marker as a size standard, the labelled side should be unified between STR alleles and the allelic ladder alleles.

Adenine↗

Two types of chicken 2',5'-oligoadenylate synthetase mRNA derived from alleles at a single locus.

We have isolated two types of chicken 2',5'-oligoadenylate synthetase cDNAs, A and B, which encode predicted proteins of 508 amino acids (58316 Da) and 476 amino acids (54336 Da), respectively. The region of A-protein comprising 33 amino acid residues from 385Ala to 417Cys is substituted by a single amino acid 385Tyr in B-protein. The homology between chicken and mammalian 2',5'-oligoadenylate synthetases is 49.5% over the amino-terminal 337 residues. Proteins expressed from A- and B-cDNAs in E. coli cells were both active in synthesizing 2',5'-oligoadenylate. However, the activity of B-protein was 10-15% of that of A-protein. Southern blotting hybridization indicated that the chicken synthetases are encoded by a single gene. RT-PCR and PCR analyses of RNA and DNA of chicken erythrocytes together with the sequence data of the PCR products showed that A- and B-mRNAs are derived from alleles at a single locus encoding chicken 2',5'-oligoadenylate synthetase, designated as OAS * A and OAS * B. Chickens carrying OAS * A/B produce two types of synthetase with molecular masses of 58 and 54 kDa, and those carrying OAS * A/A produce only a single type of 58 kDa.

2',5'-Oligoadenylate Synthetase↗

Natural killer cell-derived large granular lymphocyte lymphoma of lung developed in a patient with hypersensitivity to mosquito bites and reactivated Epstein-Barr virus infection.

A 17-year-old female developed natural killer (NK) cell-derived large granular lymphocyte (LGL) lymphoma of the lung. She had a past history of hypersensitivity to mosquito bites (HMB). After an eight-year chronic, active Epstein-Barr virus (EBV) infection, she developed multiple lung lesions and pleural effusion. In the effusion, 60% of the cells were LGL. They were CD2+, 3-, 16+, 56+, 57+, 45RO+/RA + weak, and possessed strong NK activity. No rearrangement of T-cell-receptor genes was detected. From all these results, a diagnosis of NK-LGL lymphoma of the lung was made. EB virus DNA was detected in cells infiltrating the pleural effusion. The clonality of the LGLs was determined by Southern blot hybridization with the terminal repeat sequence of EB virus as a probe, and by chromosomal abnormalities. The patient died from respiratory failure. Necropsy of the lung revealed diffuse lymphoma composed of polymorphic cells with typical angiocentric lesions. Reportedly, lymphomas of NK lineage show predominantly extranodal involvement, and primary lung lesions are rare. In the pleural effusion of the present case, abnormally high levels of soluble Fas ligand, interleukin-10 and interferon gamma were detected. This hypercytokinemia, reflecting the microenvironment of lymphoma cells, may play a role in the progression of the lymphoma and organ injury in the lung.

Adolescent↗

Age-related degeneration of the serotoninergic fibers in the zitter rat brain.

The serotonin neuron system was studied using immunohistochemical and neurochemical techniques in zitter mutant rats aged 1-14 months, which are characterized by abnormal metabolism of superoxides. The morphology of the serotoninergic neuron system and the serotonin level in the zitter rat were compared to those of age-matched Sprague-Dawley (SD) rats. Up to age 4 month, the density and distribution of serotoninergic fibers in the zitter rat brain were similar to those of control rats. However, several serotoninergic fibers with abnormal morphology, characterized by swollen varicosities, were observed in the cerebral cortex and caudate putamen of 6-month-old zitter rats. The density and distribution of these fibers in other regions of the brain were similar to those of control rats. The abnormal serotoninergic fibers increased in number and extended into other regions of the brain such as the thalamus, hippocampus, and vestibular nucleus. On the other hand, the density of normal serotoninergic fibers decreased throughout the brain of the 14-month-old zitter rat. Abnormal serotoninergic fibers have also been reported in the brain of normal older (24 months) SD rats. Neurochemical analysis revealed lower levels of serotonin, and 5-hydroxyindoleacetic acid, in all cortical areas (prefrontal, parietal, and occipital cortices), the caudate putamen, and the hippocampus of 12-month-old zitter rats. Levels differed significantly in the parietal cortex and hippocampus between the zitter and SD rats. Based on the morphological and neurochemical similarities, the present results suggest that age-related degeneration of serotoninergic fibers occurs in the zitter rat brain. Furthermore, degeneration of serotoninergic fibers appears to be induced by superoxide species. Thus, the zitter rat may provide a good model for studying the neurotoxic effects of superoxide species on the serotoninergic neuron system.

Aging↗

Frequent clonal expansion of peripheral T cells in patients with autoimmune diseases: a novel detecting system possibly applicable to laboratory examination.

To investigate T cell involvement in antigen-specific immune responses, it is important to detect accumulating T cells at a clonal level in vivo. However, thus far the clinical application of such analyses has been limited. Here we have established novel primers to anneal with T cell receptor (TCR) beta genes of multiple Vbeta families and applied them to reverse transcription-polymerase chain reaction-single strand conformation polymorphism (RT-PCR-SSCP) analysis to evaluate peripheral T cell clonality of autoimmune disease patients. As a result, the new Vbeta primers could detect accumulating T cell clones in the periphery of healthy individuals and patients. It was revealed that patients with autoimmune diseases such as systemic lupus erythematosus (SLE) had a larger number of clonal accumulations of peripheral T cells compared with normal individuals. Thus, the RT-PCR-SSCP system using the new multifamily Vbeta primers is the first such laboratory examination to detect T cell clonal expansion, and will provide a simple and sensitive tool to aid in the diagnosis and also in the investigation of the pathogenesis of autoimmune diseases.

Adult↗

Cough-challenge trial with a new angiotensin-converting enzyme inhibitor, imidapril.

This study was conducted to examine whether imidaprilat, an active diacid of the angiotensin-converting enzyme (ACE) inhibitor imidapril, preferentially inhibits angiotensin I degradation rather than bradykinin degradation, and whether imidapril is less active than other ACE inhibitors in inducing cough in patients with hypertension. The effect of imidaprilat on the inhibition of pressor response to angiotensin I and augmentation of depressor response to bradykinin was compared with that of enalaprilat and captopril in anesthetized rats. To determine the incidence of cough associated with imidapril, patients with a history of ACE inhibitor-induced dry cough were enrolled in a randomized, open-labeled, crossover trial with two 6-week periods to be treated with imidapril or amlodipine, a calcium-channel blocker. The recurrence of cough was assessed during both treatments. In the animal study, there were no significant differences in the ratio of inhibition of pressor response to angiotensin I and the augmentation of depressor response to bradykinin among the ACE inhibitors. In the cough-challenge trial, a total of 60 patients with hypertension were enrolled in the study. Cough and cough related symptoms recurred in 98.3% of the patients (59/ 60) during imidapril therapy. In contrast, only two patients reported cough during treatment with amlodipine. These results indicate that imidapril has no selectivity in inhibiting angiotensin I- and bradykinin-degradation in rats, and that clinically it is not different from other ACE inhibitors in inducing cough in patients with hypertension.

Angiotensin-Converting Enzyme Inhibitors↗

Function of aberrant EGFR in malignant gliomas.

The most common alteration of the epidermal growth factor receptor (EGFR) gene in human malignant gliomas is an in-frame deletion of exon 2-7 from the extra-cellular domain. To study the relationship between the expression of this aberrant EGFR and cell proliferation, as well as apoptosis in malignant gliomas, we have developed U-87MG cell transfectants that express the aberrant (mutant-type) or normal (wild-type) EGFR. We analyzed cell number, tumor volume, and MIB-1 positive rate as proliferation markers, and found that in tissue culture, tumors derived from U-87 MG cells (mutant-type) have the same proliferative activity as those derived from U-87 MG cells (wild-type). However, when cells expressing mutant EGFR were implanted into nude mice subcutaneously, the tumorigenic capacity was much enhanced. We also found that the apototic index of tumors derived from U-87 MG cells (mutant-type) was less than 0.1%, whereas that of wild-type tumor was 1%. These results suggest that aberrant EGFR affects the malignancy of glioma by stimulating proliferation and inhibiting apoptosis.

Animals↗

Direct measurement of acetylcholine release in detrusor smooth muscles isolated from rabbits.

In the present study, we measured acetylcholine (ACh) released from rabbit detrusor smooth muscle strips induced by electrical field stimulation (EFS) using high-performance liquid chromatography coupled with microdialysis procedure. There were frequency- and duration-dependent increases in contractile response and ACh release. There was a significant, but not simple correlation between EFS-induced contraction and ACh release. Atropine caused a decrease and increase in the contractile response and ACh release, respectively. Pretreatment with propranolol increased ACh release, but pretreatment with phentolamine had no significant effect. These results demonstrate that this method is applicable to direct measurement of ACh release by EFS, and that neurotransmitters other than ACh may relate to EFS-induced contraction. In addition, it is suggested that there are prejunctional inhibitory muscarinic receptors and beta-adrenoceptors, which contribute to ACh release induced by EFS in the rabbit detrusor smooth muscles.

Acetylcholine↗

Transumbilical resection and umbilical plasty for patent omphalomesenteric duct.

This paper describes a transumbilical resection and umbilical plasty for treatment of a patent omphalomesenteric duct (POMD). In a newborn infant with a POMD, a skin incision was made circumscribing the mucocutaneous junction of the protruded duct. The duct was completely extirpated with a wedge resection of the connection to the intestine and an umbilical plasty was performed. The postoperative appearance was excellent. It appears that transumbilical resection and umbilical plasty may be a satisfactory operation for POMD.

Humans↗

Ileal atresia associated with an omphalomesenteric duct remnant.

This paper describes an unusual case of ileal atresia associated with an omphalomesenteric duct (OMD) remnant. A 1-day-old male developed bilious vomiting and abdominal distension. At laparotomy, the distal portion of the ileum was connected to the umbilicus with a fibrous band. Around this omphalomesenteric band, the ileum was strangulated and necrotic; the end of the ileum was atretic. Apart from these structures, the cecum terminated without mesentery. This experience shows evidence of an OMD remnant as a possible cause of jejunoileal atresia.

Humans↗

Portal hypertensive enteropathy in biliary atresia.

This paper describes a rare case of biliary atresia (BA) in which massive postoperative bleeding developed due to portal hypertensive enteropathy. A 15-year-old boy had been doing well after Kasai's operation for BA. At around the age of 6 years he developed recurrent episodes of esophageal variceal bleeding and underwent esophageal transection, splenectomy, sclerotherapy, and embolization of the left gastric vein. At the age of 15 years he suddenly developed abdominal pain, hematemesis, and massive tarry stools. His hemoglobin level was 6.3 g/dl. Endoscopy showed several small, polypoid lesions in the jejunum beyond Treitz' ligament. The source of the bleeding was one of these lesions located in the proximal jejunojejunostomy of the Roux-en-Y loop. Because he had repeated episodes of melena, he underwent partial resection of the jejunum under endoscopic guidance. He has since been free of gastrointestinal (GI) bleeding. From this experience, we conclude that the polypoid lesions of an enteropathy may be a cause of massive GI bleeding in the postoperative portal hypertension of BA.

Adolescent↗

Vulnerability to aging in the rat serotonergic system.

Distributional changes of serotonergic fibers associated with aging were demonstrated immunohistochemically. Old rat brains were morphologically characterized by the presence of peculiar features of serotonergic fibers not found in the young adult brain. In 24-month-old rats, these aberrant serotonergic fibers were subdivided into two groups according to morphological alterations: type 1 fibers consisting of thin fibers with moderately enlarged varicosities, and type 2 fibers consisting of much thicker fibers that have even larger varicosities and a tortuous course. These two types of fibers were distributed differentially in the forebrain. Type 1 fibers were found mainly in the striatum and frontoparietal cortex, whereas type 2 fibers were found in the posterior cingulate cortex and dentate gyrus of the hippocampal formation. Both types of aberrant fibers were seen in amygdala, frontoparietal cortex, hypothalamus, and thalamus. In 36-month-old rats, more highly degenerating arborizations were detected, and these aberrant ramifications were classified as follows based on shape as: type 3 fibers consisting of highly arborized thin fibers with a larger number of larger varicosities, and type 4 fibers consisting of thick fibers with abundant larger varicosities. Distributional difference indicated that type 1 fibers progress into type 3 fibers, whereas type 2 develop into type 4 fibers. These findings suggest the possibility that one set of pathological fibers emanate from the dorsal raphe nucleus and the other from the median raphe. Moreover, both two sets of serotonergic fibers show age-related aberrations in their morphology over same time course.

Aging↗

Colonoscopic diagnosis of lymphoid hyperplasia causing recurrent intussusception: report of a case.

This paper describes a 6-year-old boy with recurrent ileocecal intussusception due to lymphoid hyperplasia in the terminal ileum, which was diagnosed preoperatively by colonoscopy. At the age of 3 years, he developed diarrhea and a tender abdominal mass. He was diagnosed as having intussusception by ultrasound and was treated by hydrostatic barium enema. After resolution, he had three recurrent episodes of intussusception. A contrast barium enema revealed a small mass in the ileocecal region. Colonoscopy showed several exaggerated folds of the terminal ileum and a biopsy showed lymphoid hyperplasia. Because the repeated intussusception seemed to have been caused by the lymphoid hyperplasia in the terminal ileum, he underwent an ileocecal resection without any subsequent recurrence. Based on the above findings, we conclude that a colonoscopy may thus be useful both for diagnosing lymphoid hyperplasia in the terminal ileum as a cause of recurrent intussusception and for deciding how to manage it.

Child↗

Slight difference in primary amino acid sequence of p17 matrix protein of HIV-1 exerts profound influence on its antigenicity.

HIV-1 p17 antigen has been studied for its biological significance in vitro as well as its immunological roles in vivo. By immunological approach of antibody-binding to HIV-1 p17 antigens of several subtypes in combination with computerized analysis of those tertial structures, it became evident that, irrelevant of similarity of linear amino acid sequence of different HIV-1 subtypes, a few amino acid substitutions close to or distant from specified epitope(s) affected their tertial structure resulting in change in ability of its binding to selected antibody. ELISA employing two monoclonal antibodies, A144 and C415, could detect p17 of subtypes A and B, but not of subtypes C, D, and E. Since the epitope site corresponding to A144 has been reported to be important for biological activity of p17 of HIV-1, change in tertial structure around this epitope may explain some difference in biology of HIV-1, such as infectivity of subtypes B and E.

Amino Acid Sequence↗

MRI signal characteristics of progressive massive fibrosis in silicosis.

AIM: To determine the magnetic resonance imaging (MRI) signal characteristics of progressive massive fibrosis (PMF) in silicosis. SUBJECTS AND METHODS: We evaluated prospectively the MR appearances in 17 patients with 34 PMF lesions on the basis of pre-contrast signal intensity (SI) and SI pattern and post-contrast enhancement pattern, using a 0.5-T unit. There were 13 PMF lesions in six patients who had silicosis and 21 PMF lesions in 11 patients who had silicotuberculosis. The SI pattern on T2-weighted image (WI) was classified into four types and the pattern of contrast enhancement on T1-WI was classified into three types. MR appearances of PMF lesions were correlated with the findings of computed tomography (CT). RESULTS: The commonest signal intensity characteristic was isointensity (70%) on T1-WI and hypointensity (68%) on T2-WI when compared with skeletal muscle. For signal pattern on T2-WI, a type with only internal high SI areas (46% in silicosis group, 38% in silicotuberculosis group) was most frequent. All of these areas corresponded to the low density areas at CT, suggestive of necrosis. After intravenous contrast medium enhancement, rim enhancement (54% in silicosis group, 52% in silicotuberculosis group) was most frequent, followed by no enhancement. CONCLUSION: The most common MRI appearance of PMF was isointensity on T1-WI and hypointensity on T2-WI when compared with skeletal muscle, with internal high SI areas on T2-WI and either rim enhancement or no enhancement.

Aged↗

Effect of the NO scavenger carboxy-ptio on endothelium-dependent vasorelaxation of various blood vessels from rabbits.

In the present study, we investigated the effect of a nitric oxide (NO) scavenger, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl 3-oxide [carboxy-PTIO], on endothelium-dependent relaxation of a series of blood vessels from rabbits, such as thoracic aorta and femoral, renal, mesenteric, and pulmonary arteries, using a functional muscle bath technique. Carboxy-PTIO produced concentration-dependent contractions in various vessels. The contractile responses in renal, mesenteric, and pulmonary arteries were significantly greater than those in the aorta and femoral artery. Similarly, phenylephrine-induced contractions in renal, mesenteric, and pulmonary arteries were markedly enhanced after pretreatment with carboxy-PTIO. Also, carboxy-PTIO inhibited acetylcholine-induced relaxation in various blood vessels. The maximum inhibitions in aorta and femoral artery were significantly greater than those in renal, mesenteric, and pulmonary arteries. The present data demonstrate that carboxy-PTIO reduces basal, phenylephrine-, and acetylcholine-induced release of NO in rabbit blood vessels. However, different degrees of inhibition of endothelium-dependent vasorelaxation were observed in various vessels. Specifically, the thoracic aorta and femoral artery are less susceptible to the action of carboxy-PTIO without acetylcholine than renal, mesenteric, and pulmonary arteries. Conversely, the most potent carboxy-PTIO-induced inhibition of acetylcholine-induced vasorelaxation was observed with aorta and femoral arteries. Thus, it is suggested that the contribution of endogenous NO to vascular tone and regional blood flow may vary among different rabbit blood vessels.

Acetylcholine↗