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Biomedical subjects

S Ueda

Publications and source records attributed to S Ueda.

At least 541 records · Page 30Linked to original sources
PubMed1992

Characterization of rabies virus glycoprotein expressed by recombinant baculovirus.

A cDNA of the glycoprotein (G protein) gene of rabies virus Nishigahara strain was cloned and inserted into a baculovirus genome under the control of the polyhedrin promoter. Infection of Spodoptera frugiperda cells with this recombinant virus produced a large quantity of new protein instead of the parental polyhedrin protein. By immunofluorescent and immunoblotting analyses, the recombinant protein was antigenically similar to the authentic G protein. Its molecular mass estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis, however, was slightly smaller than that of the authentic one, and this observation was suggested to be due to the difference in glycosylation level between the two G proteins. The recombinant G protein expressed on the cell surface of the insect cells showed a fusion activity at low pH. The fusion activity was inhibited by antiserum against either whole virions or G protein of rabies virus.

Animals↗
PubMed1992

Concerted evolution of the primate immunoglobulin alpha-gene through gene conversion.

We determined four nucleotide sequences of the hominoid immunoglobulin alpha (C alpha) genes (chimpanzee C alpha 2, gorilla C alpha 2, and gibbon C alpha 1 and C alpha 2 genes), which made possible the examination of gene conversions in all hominoid C alpha genes. The following three methods were used to detect gene conversions: 1) phenetic tree construction; 2) detection of a DNA segment with extremely low variability between duplicated C alpha genes; and 3) a site by site search of shared nucleotide changes between duplicated C alpha genes. Results obtained from method 1 indicated a concerted evolution of the duplicated C alpha genes in the human, chimpanzee, gorilla, and gibbon lineages, while results obtained from method 2 suggested gene conversions in the human, gorilla, and gibbon C alpha genes. With method 3 we identified clusters of shared nucleotide changes between duplicated C alpha genes in human, chimpanzee, gorilla, and gibbon lineages, and in their hypothetical ancestors. In the present study converted regions were identified over the entire C alpha gene region excluding a few sites in the coding region which have escaped from gene conversion. This indicates that gene conversion is a general phenomenon in evolution, that can be clearly observed in non-functional regions.

Animals↗
PubMed1992

Adenomyomatosis of the gallbladder: a clinical survey of 30 surgically treated patients.

Thirty patients with adenomyomatosis of the gallbladder (AMG) were operated on between January 1983 and June 1990. They were made up 3.3% of patients who underwent cholecystectomy during the same interval. Of the 30 patients, ages ranged from 22 to 77 years (mean 52.3 years) and the male-to-female ratio was 8:7. Among the macroscopic types, 10 cases of generalized, 12 of segmental (S) and 8 of fundal (F) were noted, and the size of the affected portion in type S (0.8 +/- 0.2 cm, mean +/- SD) was significantly thinner than in other two types (p less than 0.05). Although the main symptom was abdominal pain, the majority of patients with type F had no complaints. Twenty patients (27%) were accompanied by gallstones including cholesterol stones in 60% of cases, and all six cases showing microbes in the bile had gallstones. Only six patients were diagnosed as AMG by preoperative imaging techniques. Other diagnoses comprised 15 of chronic cholecystitis and 3 of suspected gallbladder carcinoma. To identify the expanded Rokitansky-Aschoff sinuses, endoscopic retrograde cholangiography and/or ultrasonography of the abdomen were most useful. No preponderant coexistent lesion other than gallstones was noted. Levels of carcinoembryonic antigen in gallbladder bile in cases of AMG (2.5 +/- 1.5 ng/ml, mean +/- SD) were significantly lower than in gallbladder carcinoma (p less than 0.01). All the patients were easily treated with cholecystectomy, and 24 patients who have been followed up after surgery are doing well.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗
PubMed1992

Type I congenital plasminogen deficiency is not a risk factor for thrombosis.

The risk of thrombosis in type I congenital plasminogen (PLG) deficiency has been suggested, but is still not confirmed. We studied 40 members of two unrelated families with this disease, and found that 21 were heterozygotes of type I congenital PLG deficiency. Three of them had thrombosis, but the other 18 had no thrombosis. The percentages of family members with no history of thrombosis up to a given age among subjects with type I congenital PLG deficiency and healthy controls were analyzed by the Kaplan-Meier method. No significant difference between the two groups was observed by the generalized Wilcoxon test (p = 0.23). These results suggest that there is no significant correlation between type I congenital PLG deficiency and thrombosis.

Adult↗
PubMed1992

Multicentre trial of ABO-incompatible kidney transplantation. Japanese Biosynsorb ABO-incompatible kidney transplant study group.

A multicentre study of ABO incompatible kidney transplantation using Biosynsorb was started in Japan in November 1989. A total of 51 cases were registered comprising 23 cases of A incompatibility, 26 cases of B incompatibility and two cases of AB incompatibility. The removal of antibodies (IgG and IgM) was carried out using Biosynsorb in 16 cases, plasmapheresis in four cases and use of both combined in 31 cases. The treatment using Biosynsorb was repeated 3.4 times on average. Serum titres of anti-A (IgG and IgM) antibodies decreased to 4.9 +/- 5.0 and 2.7 +/- 1.7 and for anti-B titres decreased to 2.8 +/- 3.5 and 2.4 +/- 3.2. Rejection was found in 33 cases: hyperacute one, accelerated acute five, and acute 27. In two cases rejection was developed concomitantly with a steep elevation in antibody titres. Three patients died, two with functioning grafts. Eight grafts were lost. Patient and graft survivals at 2 years were 94.1% and 84.3%, respectively. From these results it is concluded that: 1. Biosynsorb and plasmapheresis are effective in removing anti-A and anti-B antibodies; 2. graft and patient survivals are similar to those in ABO compatible cases; 3. anti-A and anti-B titres less than 16 are recommended at the time of transplantation; 4. anti-A and anti-B titres higher than 128 may be considered as a risk factor for rejection in the early stages after transplantation.

ABO Blood-Group System↗
PubMed1992

The DNA sequence of gamma-glutamyltranspeptidase gene of Bacillus subtilis (natto) plasmid pUH1.

The gamma-glutamyltranspeptidase (gamma-GTP) gene of Bacillus subtilis (natto) plasmid designated pUH1, which is responsible for polyglutamate production, has been cloned and the nucleotide sequence determined. The sequence contains a single open-reading frame stretching for 1260 bp with a relative molecular mass of 49,356. Putative -35 and -10 sequences, TTCAAA and TATTAT, were observed as the consensus sequence for the promoter recognized by the sigma 43 RNA polymerase of B. subtilis, and the ribosome binding site, the sequence of which was AACGAG, was complementary to the binding sequence of B. subtilis 16S rRNA except for one base. The amino acid sequence of the gene with the segment of putative protein C403 of staphylococcal plasmid pE194 indicates homology, whereas that with Escherichia coli and mammalian gamma-GTPs does not show any similarity at all.

Amino Acid Sequence↗
PubMed1992

Membrane capacitance increases induced by histamine and cyclic AMP in single gastric acid-secreting cells of the guinea pig.

During acid secretion, gastric parietal cells undergo profound morphological changes including formation of the apical secretory membrane. To examine the mechanism of histamine-induced increases in the apical membrane area at the single cell level, we monitored the membrane capacitance by applying a time-resolved phase-sensitive detection method to singly isolated parietal cells of guinea pig. A real-time increase in the membrane capacitance was detected within several min after stimulation with histamine. An H2-blocker (cimetidine), but not an H1-blocker (pyrilamine), inhibited the histamine response. Dibutyryl cyclic AMP mimicked the histamine effect. The capacitance response to histamine was sensitive to cytosolic Ca2+, temperature and N-ethylmaleimide. The histamine response was inhibited by intracellular application of a non-hydrolyzable ATP analog (AMP-PNP) and an isoquinolinesulfonamide derivative that works as an inhibitor of protein kinase A (H-8). These results indicate that in parietal cells, elevation of intracellular cyclic AMP induces exocytotic insertion of intracellular membranes into the plasma membrane, presumably by activating protein kinase A.

Animals↗
PubMed1992

Pathological study of diffuse axonal injury patients who died shortly after impact.

It is generally considered that axonal injury is apparent only on electron microscopy in the very early stage after a closed head injury. To clarify the pathological findings in head injury patients dying very shortly after the impact, we analyzed 8 fatal cases of diffuse axonal injury (DAI) who underwent medicolegal autopsy at the Department of Forensic Medicine of Kyoto Prefectural University of Medicine. Seven cases died within one hour after injury and another one case died 3 days after injury. We studied these cases macroscopically, microscopically, and electron microscopically. Macroscopically all cases showed the typical findings of diffuse axonal injury. Microscopical study of the cases who died within one hour revealed no characteristic findings of DAI such as appearance of retraction balls or microglia. On the other hand, in the case who died only 3 days after injury it showed the typical retraction balls. Electron microscopic study showed the remarkable destruction of cytoskeletal structure of axons in all cases. From our results, it is reasonable to speculate that DAI may be common among head injury patients who die very soon after the impact.

Adolescent↗
PubMed1992

Effects of a platelet-activating factor antagonist, CV-6209, on gastric mucosal lesions induced by ischemia-reperfusion.

Recent research was shown that oxygen-derived free radicals are involved in the pathogenesis of various diseases, including ischemia-reperfusion injury. We have also reported that oxygen-derived free radicals and lipid peroxidation may play an important role in gastric mucosal injury induced by ischemia-reperfusion. The hypoxanthine-xanthine oxidase system and neutrophils are considered important sources of oxygen-derived free radicals in this process. In recent years, it also has been shown that serum platelet-activating factor (PAF) levels increased during ischemia-reperfusion, and that induction of superoxide generation by neutrophils is one of the important biological effects of PAF. In the present study, we examined the effect of CV-6209, a specific PAF receptor antagonist, on gastric mucosal injury induced by ischemia-reperfusion, to shed some light on the possible involvement of PAF in such lesions. CV-6209 significantly attenuated the gastric mucosal injury induced by ischemia-reperfusion, and inhibited both an increase of thiobarbituric acid reactive substances and a decrease of alpha-tocopherol in gastric mucosa after ischemia-reperfusion. However, CV-6209 had no effect on gastric mucosal blood flow during ischemia-reperfusion. These results suggest that endogenous PAF may play an important role in gastric mucosal injury induced by ischemia-reperfusion, and that CV-6209 exerts its beneficial effect mainly by inhibiting neutrophil superoxide production induced by PAF.

Animals↗
PubMed1992

Pre- and postnatal stimulation of pulmonary surfactant protein D by in vivo dexamethasone treatment of rats.

Fetal (days 18 and 20 of gestation), neonatal (days 0, 2 and 4 of neonate) and adult rats were injected with dexamethasone (1 mg/kg) in vivo and 24 hours later the effect on the contents of surfactant protein D (SP-D) in the rat lungs were examined in comparison with surfactant protein A, disaturated phosphatidylcholine and phosphatidylglycerol. In vivo dexamethasone treatment resulted in significant increases of SP-D content as the other 3 components of surfactant in both fetuses and neonates, but not in adults. Responsiveness to glucocorticoid treatment on SP-D content was maximum on day 1 of neonate (2.7 times control value). The contents of surfactant components examined tend to respond better to steroid in postnatal rats. These data demonstrated that glucocorticoid treatment in vivo for short durations exhibits the stimulatory effect on the contents of SP-D in the fetal and neonatal rat lungs.

Aging↗
PubMed1992

Sequence determination of cDNA clones of transcripts from the tumor-associated region of the Marek's disease virus genome.

The number of 132-bp tandem direct repeats within the long inverted repeat region of the Marek's disease virus type 1 (MDV1) genome increases concomitantly with the loss of oncogenicity during serial passages in cultured cells. Twelve clones carrying the 132-bp sequence were isolated from a cDNA library constructed from chicken embryo fibroblasts infected with the MDV1 Md5 strain. Through sequence analysis of a cDNA clone and primer extension analysis, the corresponding mRNA was found to be a linear transcript which included the two 132-bp tandem direct repeats. Two open reading frames were found in this transcript. One had a week homology with v-fms. The other should increase its size concomitantly with expansion of the 132-bp tandem direct repeat. PCR analysis of both cDNA clones and RNA gave amplified products which were as large as that produced from the genomic clone, indicating that a majority of mRNA from this region is composed of unspliced transcripts.

Amino Acid Sequence↗
PubMed1992

Immunohistochemical and enzymehistochemical studies of peptidergic, aminergic and cholinergic innervation of the lacrimal gland of the monkey (Macaca fuscata).

The distribution of nerve fibers containing peptides which include calcitonin gene-related peptide (CGRP), substance P (SP), neuropeptide Y (NPY) and vasoactive intestinal polypeptide (VIP) and enzymes of tyrosine hydroxylase (TH) and acetylcholinesterase (AchE) in the lacrimal gland of the monkey (Macaca fuscata) was studied using immunohistochemical and enzymehistochemical methods. We also examined the trigeminal ganglion (TG) and superior cervical ganglion (SCG) using the same methods. All peptide- and enzyme-containing nerve fibers examined in this study were present in the lacrimal gland and a consistent distribution pattern for each substance was found. CGRP-immunoreactive (IR) nerve fibers were mainly distributed around the blood vessels in the interlobular connective tissue. The distribution pattern of SP-IR nerve fibers was similar to that of CGRP-IR nerve fibers, but they were much less in number. NPY-IR nerve fibers were observed mostly around the blood vessels and occasionally in the interstitial stroma between the acini. Numerous VIP-IR nerve fibers were found surrounding the acini, ducts and blood vessels. TH-IR nerve fibers were also been around the blood vessels and in the interstitial stroma between the acini, as were NPY-IR fibers. The highest concentration of acetylcholinesterase (AchE)-positive nerve fibers was present in the acini, ducts and blood vessels, showing a similar distribution to VIP-IR fibers. In the TG, 50% of medium and 30% of small ganglion cells were CGRP-IR cells, while 20% of medium and 25% of small ganglion cells were of the SP-IR types.(ABSTRACT TRUNCATED AT 250 WORDS)

Acetylcholinesterase↗
PubMed1992

Augmentative effects of tumor necrosis factor-alpha (human, natural type) on polymorphonuclear leukocyte-derived superoxide generation induced by various stimulants.

We investigated the effects of tumor necrosis factor-alpha (human, natural type: n-TNF) on polymorphonuclear leukocyte (PMN)-derived superoxide generation by the new method of Cypridina luciferin analog-dependent chemiluminescence, which had high sensitivity and specificity to superoxide. Preincubation of PMNs with n-TNF for 3 min increased PMN-derived superoxide generation induced by phorbol myristate acetate, A23187, opsonized zymosan and N-formyl-methionyl-leucyl-phenylalanine in a concentration dependent manner (0.5-50 Japan reference units/ml of n-TNF). In addition, the enhanced PMN-derived superoxide generation by n-TNF showed a positive correlation to the preincubation time of PMNs with n-TNF (3-15 min). However, a direct incubation of PMNs with n-TNF for 1 h did not induce superoxide from PMNs without the above stimulants. The augmentative effects of n-TNF on PMN-derived superoxide generation should be useful in the PMN-mediated host defense mechanism, such as bactericidal and antitumor activity. The local concentration of n-TNF and the n-TNF-PMN contact time are considered very important in obtaining these effects more efficiently in addition to the presence of PMN-stimulants including complements, chemotactic peptides and phorbol esters.

Dose-Response Relationship, Drug↗
PubMed1992

Nasal antibody response to mumps virus after vaccination and natural infection.

Nasopharyngeal secretions and sera were collected during an epidemic of mumps in a semi-closed institution. None of the children who received live attenuated mumps vaccine 5 months previously developed any symptoms of mumps. Neutralizing and IgA antibodies against mumps virus were measured by the peroxidase-antiperoxidase method and the fluorescent antibody to membrane antigen method, respectively. The immune responses in nasopharyngeal secretions and serum were studied in four groups (apparently infected, non-apparently infected, non-infected and vaccinated). We found that IgA, which has neutralizing activity, was produced after vaccination as well as after natural infection. It is suggested that local antibody induced by vaccination may contribute to protection against the mumps virus.

Antibodies, Viral↗
PubMed1992

Immunoglobulin CH gene family in hominoids and its evolutionary history.

The organization of the human immunoglobulin CH gene suggests that a gene duplication involving the C gamma-C gamma-C epsilon-C alpha region has occurred during evolution. We previously showed that both chimpanzee and gorilla have two 5'-C epsilon-C alpha-3', as in human, and that orangutan, gibbon, and Old World monkeys have one C epsilon gene and one, two, and one C alpha gene(s), respectively. In addition to these clustered CH genes, there is one processed C epsilon pseudogene in each species. The present study revealed that orangutan and crab-eating macaque (an Old World monkey) both have one 5'-C epsilon-C alpha-3' and that gibbon has two 5'-C epsilon-C alpha-3', one C epsilon gene of which is completely deleted. By Southern analysis, the number of C gamma genes in all the nonhuman hominoids was estimated to be four to five, as in human, in comparison with two for crab-eating macaque. The C mu and C delta genes were estimated to be present as single copies in both hominoids and crab-eating macaque. Furthermore, it was proved that there are two copies of the C epsilon 5'-flanking region in both the orangutan and the gibbon genomes. These results show that gene duplication including the C gamma-C gamma-C epsilon-C alpha genes occurred in the common ancestor of hominoids and that subsequent deletion of the C epsilon gene (in orangutan, including one of the C alpha genes) occurred independently in each hominoid species.

Animals↗
PubMed1992

Selective amplification of cDNA sequence from total RNA by cassette-ligation mediated polymerase chain reaction (PCR): application to sequencing 6.5 kb genome segment of hantavirus strain B-1.

A method, referred to as cassette-ligation mediated polymerase chain reaction (PCR), has been developed to permit selective and specific amplification of cDNA sequence from total cellular RNA. This technique comprises (i) digestion of cDNA with multiple restriction enzymes, (ii) ligation of cleavage products to double-stranded DNA cassettes possessing a corresponding restriction site and (iii) amplification of cassette-ligated restriction fragments containing a short, known sequence (but not all the other ligation products) by PCR using the specific and cassette primers; the specific primer is designed to prime synthesis from the known sequence of the cDNA whereas the cassette primer anneals to one strand of the cassette. Sequencing from the cassette primer provides information to design a new primer for the next walking step. The amplified cDNA fragments are often larger than the maximum DNA fragments (500-600 bp) that can be sequenced without the need of synthesizing internal sequencing primer. Each of such large cDNA fragments is dissected into smaller DNA fragments by repeating cassette-ligation mediated PCR exploiting different restriction sites and different sets of cassette primers. This dissection process reduces the number of specific primers to a minimum, thereby increasing the speed of sequencing and minimizing the overall cost. We have successfully applied this cDNA walking and sequencing by the cassette-ligation mediated PCR to the sequencing of an entire 6.5 kb genome segment of hantavirus strain B-1.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1992

Estrogen as a growth factor to central nervous cells. Estrogen treatment promotes development of acetylcholinesterase-positive basal forebrain neurons transplanted in the anterior eye chamber.

In a previous report, we demonstrated in vivo ameliorating effects of conjugated estrogen in women suffering from senile dementia-Alzheimer's type. To investigate the effects of estrogen on the growth of cholinergic neurons, the present study was performed using rat cholinergic tissue implanted into the anterior chamber of the eye. Fetal diagonal band tissue containing cholinergic neurons was grafted into the anterior eye chamber of adult female rats that had either been treated or not with 2 mg estradiol valerate injected every 3 days after oophorectomy. Two and four weeks after transplantation, the axonal and/or dendritic growth of cholinergic neurons in the graft was studied using acetylcholinesterase histochemistry. At both times, acetylcholinesterase positive processes were densely distributed in the grafts of estradiol valerate treated rats, while in rats without estradiol valerate treatment acetylcholinesterase positive reaction was essentially localized only on the cell bodies. These findings were more obvious at 2 weeks after transplantation than at 4 weeks. These results suggest that estrogen acts on cholinergic neurons as a growth factor.

Acetylcholinesterase↗