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Biomedical subjects

T Kudo

Publications and source records attributed to T Kudo.

At least 451 records · Page 25Linked to original sources
PubMed1991

A simple and improved method to generate human hybridomas.

By improving our previous method for production of human hybridomas, we developed a simple and remarkably efficient method for production of human hybridomas. We reformed our previous method in the following three points: (1) we added irradiated (30 Gy) myeloma cells as feeder cells to culture of fusion cells; (2) ouabain concentration in the selective medium was reduced from 2 x 10(-6) M to 1 x 10(-6) M; (3) selective medium (GIT-HAT-OL) was added to the culture after overnight cultivation of fusion cells. Consequently, we obtained higher fusion frequency (1/700 vs. 1/5500) compared with our previous method. By our present method, only so small number of human B cells (EBV-LCL) is required, so that the time necessary to establish human hybridomas is reduced.

Cell Fusion↗
PubMed1991

An immunohistochemical study of MAP2 and clathrin in gerbil hippocampus after cerebral ischemia.

Changes in MAP2 and clathrin immunoreactivity were studied in gerbil hippocampus after transient cerebral ischemia. MAP2 immunoreactivity decreased significantly by 1 h in the subiculum-CA1 and CA2 areas which correspond to reactive change, while no decrease was observed in CA1 until day 4. Before the initiation of delayed neuronal death, MAP2 immunoreactivity was not changed in CA1. On the other hand clathrin immunoreactivity increased in the pyramidal cell layer of CA1 by 3 h after ischemia and remained high for 2 days. Clathrin immunoreactivity in the pyramidal cell layer of CA1 diminished after delayed neuronal death. The transient change of clathrin was noted especially in CA1 in the period prior to delayed neuronal death. These results imply an abnormal change in clathrin turnover after ischemia, which may participate in the pathogenesis of delayed neuronal death.

Animals↗
PubMed1991

Buffy coat from families of Alzheimer's disease patients produces intracytoplasmic neurofilament accumulation in hamster brain.

Buffy coat of three members from a family with Alzheimer's disease was inoculated into hamster brains. Eighteen months after the inoculation, all experimental animals were sacrificed for the neuropathological study. Hematoxylin-eosin staining showed no gross vacuolar degeneration, or neuronal loss in the cortex. The spongiform degeneration was minimum. Immunostaining with antibodies against neurofilament 200 kDa subunit protein revealed massive immuno-positive intracytoplasmic inclusion bodies within neurons of the brainstem nuclei. By electron microscopy, the intracytoplasmic inclusion body was shown to be composed of proliferated 10 nm neurofilaments. The intra-cytoplasmic neurofilament proliferation was observed with the hamsters inoculated with the buffy coat from Alzheimer's disease patients as well as an apparently normal member of the family.

Alzheimer Disease↗
PubMed1991

Serum hepatitis C virus sequences in posttransfusion non-A, non-B hepatitis.

We investigated 17 patients (12 males and 5 females, ages 2 to 57 years old) with posttransfusion non-A, non-B hepatitis to determine relationships between clinical courses and hepatitis C virus (HCV) markers. The patients were grouped according to time course of abnormal serum alanine aminotransferase (ALT) levels into three categories (chronic biochemical disease, biochemically resolved chronic disease, and acute disease). Latest serum samples (1.3 to 10.8 years after blood transfusion) were used to detect antibodies against C100-3 antigen (anti-HCV) by enzyme-linked immunosorbent assay and HCV sequences by polymerase chain reaction (PCR) assay. Of the 17 patients, 13 patients (76.5%) were anti-HCV positive and 8 patients (47.1%), including one anti-HCV negative case, were positive for HCV RNA. In total, 14 patients (82.4%) were positive for either HCV markers. With respect to clinical course, HCV RNA was detected in six of eight patients (75%) with chronic biochemical disease, and in two of five patients (40%) with biochemically resolved chronic disease. HCV RNA was not detectable in convalescent sera from four patients with acute disease. These results show that there is a relationship between clinical status and HCV viremia, but that normal liver function tests do not always represent the clearance of the virus. Viremia in two patients with normal ALT level suggests that hepatitis is not only caused by viral cytopathic effects, but also by immunologic reactions against virus-infected cells. Thus, PCR is useful in determining the persistence of HCV infection as well as to diagnose anti-HCV negative HCV infection.

Adolescent↗
PubMed1991

Construction and characterization of the chimeric enzymes between the Bacillus subtilis cellulase and an alkalophilic Bacillus cellulase.

The amino acid sequences of cellulase from Bacillus subtilis (BSC) and that from an alkalophilic Bacillus sp. N-4 (NK1) show significant homology in most parts except for the C-terminal portions. Despite the high homology, the pH activity profiles of the two enzymes are quite different; BSC has its optimum pH at 6-6.5, whereas NK1 is active over a broad pH range from 6 to 10.5. In order to identify the structural features which determine such pH activity profiles, chimeric cellulases between BSC and NK1 were constructed using four restriction sites commonly present within the homologous coding sequences, and were produced in Escherichia coli. The chimeric cellulases showed various chromatographic behaviors, reflecting the origins of their C-terminal regions. The pH activity profiles of the chimeric enzymes in the alkaline range could be classified into either the BSC or NK1 type mainly depending on the origins of the fifth C-terminal regions. In the acidic range, the profile was determined only by the origin of the fourth enzyme region from the N terminus. Comparison of the kinetic parameters between pH 5 and 6 using p-nitrophenyl cellobioside as a substrate indicated that the fourth region is responsible for the pH-dependent change of the kcat value. Only a limited number of amino acids in the fourth region may affect on deprotonation of catalytic residues of the cellulases and modulate the catalytic activity in the acidic pH values.

Amino Acid Sequence↗
PubMed1991

Network system: the integrated picture archiving and communication system with the hospital information system.

We describe the outline of Hokkaido University picture archiving and communication system (HU-PACS) and its network functions. HU-PACS processes 0.5 Gbytes of images daily through 10 acquisition devices. Functional integration of hospital information system (HIS) and PACS has been implemented. The HU-PACS can access common data base with HIS. Multi-image data-base systems provide fast throughput of image retrieval. The system is composed of 4 modules: (1) 2 image data-base systems (IDS), (2) 1 image management system (IMS), (3) image display terminals (IDT), and (4) 8 image acquisition nodes interfaced to 10 modalities and 1 film scanner. These 4 modules are connected by branch and loop type local area networks (LAN). HU-PACS has functions other than basic PACS functions. Images in optical disk library (ODL) are loaded onto magnetic disks (MD) in advance according to patient booking information, the images expected to be viewed are transferred to the local storage automatically, and the desired images can be pre-downloaded into local storage by instruction through HIS terminals.

Computer Communication Networks↗
PubMed1991

Plasma vasopressin response to contrast medium during cardiac catheterization in infants and children.

Fifteen infants and children (M = 7, F = 8), aged from 0 to 13 years, who underwent cardiac catheterization and cardioangiography under ketamine-diazepam anesthesia were the subjects of this study. The effect of a contrast medium, isolamate sodium (66.8%) on the plasma somolality and vasopressin concentration was studied. The plasma osmolality was significantly elevated after contrast medium administration (289 +/- 3 vs. 303 +/- 8 mosmol.kg(-1)) as well as plasma vasopressin (from 2.1 +/- 0.9 vs. 4.7 +/- 2.0 micro-unit.ml(-1)). It is concluded that the administration of contrast medium for cardioangiography causes elevation of plasma osmolality, which leads to the elevation of plasma vasopressin concentration.

Journal Article↗
PubMed1991

Subcellular distribution of enkephalin precursor proteins in rat dental pulp and the usefulness as a substrate for enkephalin-producing enzymes.

The subcellular distribution of enkephalin (EK) precursor proteins was investigated to clarify the intracellular site of biosynthesis of EK in rat dental pulp tissue. The contents of met-EK-like peptides in nuclear, microsomal, and supernatant fractions of the pulp tissue were markedly increased after sequential digestion with trypsin and carboxypeptidase B, indicating the enrichment of the precursors in these fractions. Sephadex G-100 gel filtration showed a common peak of the precursor proteins in the homogenate and its microsomal and supernatant fractions, and the molecular weight was determined to be about 58,000 by SDS polyacrylamide gel electrophoresis. Both the partially purified precursor protein from the supernatant fraction and N alpha-benzoyl-DL-arginine-beta-naphthylamide (BANA) were used as substrates for a lysosomal enzyme separated by Sephadex G-75 gel filtration. The major peak of EK-producing activity of the enzyme was identical with that of BANA-degrading activity of the enzyme. These results demonstrate the EK-producing activity of lysosomal proteinase, and also indicate the usefulness of the two substances as substrates for the enzyme.

Animals↗
PubMed1991

Inactivation of kanamycin A by phosphorylation in pathogenic Nocardia.

Among the five species of pathogenic Nocardia, i.e., N. asteroides, N. brasiliensis, N. farcinica, N. nova and N. otitidiscaviarum, all strains of N. brasiliensis and N. farcinica showed resistance to an aminoglycoside antibiotic, kanamycin A, showing the MIC (minimum inhibitory concentration) values of more than 100 micrograms/ml. This species-specific difference in sensitivity was found to be explained by the production of an inactivation enzyme, aminoglycoside 3'-phosphotransferase APH(3'). Structural studies by mass and NMR spectroscopy on the inactivated substance produced by a cell-free extract of the Nocardia confirmed the conversion of kanamycin A to an inactive substance, kanamycin A 3'-phosphate. The MIC values of N. otitidiscaviarum and N. nova for kanamycin A, on the other hand, ranged from 0.78 micrograms/ml to 100 micrograms/ml, and both species were non-producers of APH(3'). Sensitivity to the antibiotic and APH(3') productivity of N. asteroides varied depending on the strain.

Carbohydrate Sequence↗
PubMed1991

Catecholamines in experimentally growth-retarded rat fetus.

This study was designed to evaluate the sympathoadrenal system in rat IUGR fetuses prepared by the Wigglesworth method. Samples were obtained on day 20 of gestation. In the amniotic fluid, concentrations of norepinephrine (NE) and particularly epinephrine (E) were significantly higher in IUGR fetuses than in normally developed fetuses (NE: p less than 0.05, E: p less than 0.01). L-Dopa and three catecholamines in the adrenal glands were decreased in IUGR fetuses. The mol ratios (L-dopa: dopamine: NE: E) were 1: 4: 35: 63 in IUGR fetuses and 1: 5: 39: 87 in normal fetuses. The ratio for epinephrine to L-dopa was significantly lower in IUGR fetuses (p less than 0.05). Adrenal phenylethanolamine-N-methyltransferase activities showed no difference between the two groups. From these results, adrenal epinephrine synthesis in IUGR fetuses was thought to be normal. Because of the depletion of adrenal epinephrine in IUGR fetuses, the plasma epinephrine concentrations were significantly lower after acute hemorrhagic shock (p less than 0.01). However, the plasma concentrations of dopamine and norepinephrine were significantly higher (dopamine: p less than 0.01, NE: p less than 0.001) in IUGR fetuses. These findings suggest that the hypersecretion of adrenal epinephrine is a protective response to chronic stress and the decrease of adrenal epinephrine may be a causative factor for the reduced reserves in the defense mechanism of IUGR fetuses.

Adrenal Glands↗
PubMed1991

The production mechanism of amniotic fluid dopamine in rats.

The production of dopamine in amniotic fluid was studied using Wistar rats. The dopamine concentrations in the amniotic fluid increased significantly between each day of gestation from day 18 to day 21, and a marked increase was observed during the last two days. The concentrations of dopamine in the fetal plasma, however, did not change with the gestational age. There was a significant increase in the fetal plasma L-dopa, which is the precursor of dopamine, without any changes in maternal plasma L-dopa levels during late gestation. Dopa decarboxylase (DDC) activity was also measured in the fetal rat kidney from gestation day 17 to day 21. DDC activity, which was low on day 17, increased with advanced gestation. DDC activity on day 21 was 0.68 nmol/mg tissue/h, about 13-fold higher than on day 17. These results suggest that fetal plasma L-dopa may be converted to dopamine by fetal kidney DDC, and that the dopamine is voided into the amniotic cavity in fetal urine.

Amniotic Fluid↗
PubMed1991

Characterization of intracytoplasmic neurofilament accumulation in hamster brain caused by Alzheimer buffy coat inoculation: comparison with experimental neurofibrillary changes produced by aluminum intoxication.

Neurofilament accumulation was produced in neuronal perikarya of the lower brain stem nuclei in the hamster brain by inoculating buffy coat from a patient with Alzheimer's disease. The abnormally proliferated neurofilament was characterized by immunostaining and electron microscopy. The intracytoplasmic neurofilament accumulation was positively stained with anti-tau and anti-ubiquitin, but not with antibody against paired helical filaments. Experimental neurofibrillary changes were produced in the rabbit brain by aluminum injection, and the proliferated neurofilaments in the rabbit brain were compared with those in the hamster brain inoculated with Alzheimer buffy coat. The results indicate that the long pathological process in the hamster brain is triggered by Alzheimer's buffy coat inoculation which results in intracytoplasmic accumulation of modified neurofilaments.

Aluminum↗
PubMed1991

Phospholipid metabolism in cardiomyopathic hamster heart cells.

We demonstrated that the activities of phosphatidylinositide-specific phospholipase C, inositol 1,4,5-trisphosphate (IP3) kinase, and IP3 phosphatase were enhanced in cardiomyopathic hamster hearts (BIO 14.6 and BIO 53.58) in comparison to control hamsters (F1b). Release of both arachidonic acid and prostacyclin was markedly enhanced by norepinephrine in the cardiomyopathic hamsters. Phospholipase C in heart has high substrate specificity to phosphatidylinositol. IP3 production was markedly enhanced in the cardiomyopathic hamsters. We also determined the intracellular calcium concentration, which was higher in BIO 53.58 hamsters than in BIO 14.6 hamsters at 5-20 weeks of age. There was no significant difference in the intracellular calcium level between F1b and BIO 14.6 hamsters at 5 weeks of age. These results suggest that phosphatidylinositol turnover stimulated by norepinephrine may produce high intracellular calcium levels in both BIO 14.6 and BIO 53.58 myocytes. In addition, in BIO 53.58 hamsters, some mechanism such as the sarcoplasmic reticulum, which controls the intracellular calcium level, may deteriorate in function. We concluded from these results that a prolonged high intracellular calcium level may lead to the death of BIO 53.58 myocytes.

6-Ketoprostaglandin F1 alpha↗
PubMed1991

Isolation of bovine coronavirus from feces and nasal swabs of calves with diarrhea.

Fecal and nasal samples were collected from 180 calves with diarrhea and 36 clinically normal co-habitants, and tested for virus using HRT-18 cell cultures derived from human rectal adenocarcinoma. A cytopathic virus was isolated from 5 fecal and 56 nasal samples obtained from diarrheic calves. All calves in which the virus was isolated from diarrheic feces were positive for virus isolation from nasal swabs. The virus was also isolated from the nasal swabs of 10 clinically normal calves that were co-habitants with diarrheic calves. Because they were morphologically similar to coronavirus, agglutinated mouse erythrocytes and serologically identical with the Nebraska calf diarrhea coronavirus, new isolates were identified as bovine coronavirus. The demonstration of viral antigens in nasal epithelial cells by a direct immunofluorescence was in close agreement with the virus isolation in HRT-18 cell cultures. This is the first report on the isolation of bovine coronavirus from newborn calves with diarrhea in Japan. The evidence that the virus was frequently isolated from nasal swabs is of great interest for understanding the pathogenesis of bovine coronavirus infection.

Adenocarcinoma↗
PubMed1991

Effect of elastase on aortic smooth muscle cell proliferation.

We studied the effects of elastase on [3H]thymidine incorporation into aortic smooth muscle cells. When elastase was added to cultured aortic smooth muscle cells, [3H]thymidine incorporation was inhibited in a dose-dependent manner in the presence of fetal bovine serum. Elastase also inhibited this incorporation in cells treated with epidermal growth factor. Epidermal growth factor (50 ng/ml) stimulated thymidine incorporation, without elastase, but with the addition of 20 units/ml of elastase the incorporation was inhibited 70%. The incorporation of thymidine into cells treated with 50 ng/ml epidermal growth factor was also inhibited 50% by a low concentration of elastase (5 units/ml). These inhibitory effects on thymidine incorporation were also observed in cells stimulated with platelet-derived growth factor. Platelet-derived growth factor (20 units/ml) markedly stimulated thymidine incorporation into cells, and elastase inhibited its activity in a dose-dependent manner. These results suggest that elastase has the potential to prevent the development of atherosclerosis by inhibiting smooth muscle proliferation.

Animals↗
PubMed1991

Effects of L-dopa or dopamine on human decidual prostaglandin synthesis.

L-Dopa and three catecholamines in the amniotic fluid before and after labor were measured to confirm the amniotic fluid catecholamine levels at the end of gestation. L-Dopa values were higher than those of three catecholamines, and dopamine which was the predominant catecholamine, rose significantly after the onset of labor. Then, to evaluate the effects of L-dopa or dopamine on prostaglandin synthesis, strips of human decidua vera obtained from fetal membranes at the time of elective cesarean sections before the onset of labor were incubated in Krebs-Ringer buffer in the presence of L-dopa or dopamine. When L-dopa was added, the net production of prostaglandin(PG)F was significantly greater than that of the control at each incubation time. On the other hand, the significant rise was observed only after 10 min of incubation for PGE2 production. Dopamine had a stimulatory effect on PGF synthesis only after 15 and 30 min of incubation, and it also stimulated the release of PGE2 at each incubation time. These results suggest that dopamine and L-dopa in amniotic fluid stimulate the production of prostaglandin by the decidua in humans.

Amniotic Fluid↗
PubMed1991

Purification and properties of a novel surface-active agent- and alkaline-resistant protease from Bacillus sp. Y.

In the course of a search for an alkaline stable protease for industrial use, an alkaline protease (protease BYA) was isolated from an alkalophilic Bacillus sp. Y, and its properties were characterized. Its optimum pH was pH 10.0-12.5, when casein was used as a substrate. In addition to the stability of protease BYA at pH 6.5-13.0, it was also very stable towards various surface-active agents, such as sodium dodecyl sulfate and sodium linear alkylbenzene sulfonate. Protease BYA was most active at 70 degrees C. The isoelectric point (pI) of protease BYA was about 10.1. Protease BYA was characterized as a serine protease because of its sensitivity to phenylmethanesulfonyl fluoride and diisopropyl fluorophosphate. The protease seems to be related to proteases of the subtilisin family, such as subtilisin BPN', subtilisin Carlsberg, and No. 221 protease.

Amino Acid Sequence↗