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Biomedical subjects

T Nagata

Publications and source records attributed to T Nagata.

At least 343 records · Page 19Linked to original sources
PubMed1994

Light microscopic lectin histochemistry in aging mouse kidney: study of compositional changes in glycoconjugates.

We report compositional changes in glycoconjugates in mouse kidney cortex due to aging, as analyzed by lectin histochemistry and Western blot analysis. Mouse kidney tissues of prenatal and postnatal ages (prenatal, 19 days of gestation; postnatal 2 and 8 days, 4 and 13 weeks, and 10 months) were fixed in 4% paraformaldehyde and cryosections were made. They were stained with 16 kinds of biotinylated lectin, followed by ABC, for light microscopy. Tissue homogenate was also examined by Western blotting for WGA, ConA, and Lotus. Changes in glycoconjugates due to prenatal and postnatal aging were detected by both lectin histochemistry and Western blotting.

Aging↗
PubMed1994

Uric acid degrading enzymes, urate oxidase and allantoinase, are associated with different subcellular organelles in frog liver and kidney.

On the basis of differential and density gradient centrifugation studies, the site of the uric acid degrading enzymes, urate oxidase and allantoinase, in amphibia was previously assigned to the hepatic peroxisomes. Using specific antibodies against frog urate oxidase and allantoinase, we have undertaken an immunocytochemical study of the localization of these two proteins in frog liver and kidney, and demonstrate that whereas urate oxidase is present in peroxisomes, allantoinase is localized in mitochondria. Urate oxidase and allantoinase were detected by immunoblot analysis in both frog liver and kidney. The subcellular localization of these two enzymes was ascertained by Protein A-gold immunocytochemical staining of Lowicryl K4M-embedded tissue. Peroxisomes in frog liver parenchymal cells and kidney proximal tubular epithelium contained a semi-dense subcrystalloid core, which was found to be the exclusive site of urate oxidase localization. Allantoinase was detected within mitochondria, but not in peroxisomes of hepatocytes or proximal tubular epithelium. No allantoinase was detected in the mitochondria of nonhepatic parenchymal cells in liver and of the cells lining the distal convoluted tubules of the kidney. These results demonstrate that, unlike rat kidney peroxisomes which lack urate oxidase, peroxisomes of frog kidney contain this enzyme. Contrary to previous assumptions, these studies also clearly establish that urate oxidase and allantoinase, the first two enzymes involved in uric acid degradation, are localized in different subcellular organelles in frog liver and kidney.

Amidohydrolases↗
PubMed1994

Observation of sleep-related breathing disorders in patients with coronary artery disease by ambulatory electrocardiogram-respiration monitoring system.

Eighty-five coronary artery patients examined using an ambulatory electrocardiogram-respiration monitoring system (AERMS) in which a respiratory sensor was strapped to the right upper abdominal wall. Apnea was defined as a cessation of abdominal wall movement lasting at least 10 sec. Sleep-related breathing disorder (SRBD) was diagnosed if at least 30 apneic episodes were observed during sleep. The cardiac events evaluated during follow-up included occurrence of sudden death, myocardial infarction and ventricular tachycardia. SRBD was detected in 9 of 85 patients (11%). There were more patients with low EF (EF < 50%) in the SRBD group than in the non-SRBD group (p < 0.01). During follow-up for a mean period of 18.4 +/- 7.6 months after ambulatory recording, four of nine (44%) patients in the SRBD group had cardiac events, compared with only four of 79 (6%) patients in the non-SRBD group (p < 0.001). Thus, coronary artery patients who were complicated with SRBD showed poor cardiac function and had a high incidence of cardiac events.

Aged↗
PubMed1994

Tissue residues of sulfadimethoxine following dietary administration to broiler-chickens.

Broilers were divided into four groups and the first group served as the control. The second, third and fourth groups were given feed containing 25, 50 and 100 mg/kg of sulfadimethoxine (SDM), respectively, for 21 days, and thereafter each group received the SDM free feed. On certain days during the experiment period, three broilers in each group were sacrificed and tissues, including blood, heart, liver, spleen, gizzard, thigh muscle, breast muscle and fat, were collected and residual SDM were determined by HPLC. Two days after withdrawal, SDM in each tissue had decreased to below the detection limit of 0.01 microgram/g.

Animal Feed↗
PubMed1994

[13-week oral toxicity study of lactitol (NS-4) in dogs followed by 4-week recovery test].

UNLABELLED: 13-week repeated dose toxicity studies were conducted on lactitol, a hepatic encephalopathy drug. In the experiment I, male and female dogs were orally treated with lactitol at doses of 0, 1.0, 2.5 and 6.25 g/kg/day for 13 weeks, followed by 4 weeks recovery period. In the experiment II, male and female dogs were orally treated with lactitol at doses of 0, 0.25, 0.50 and 1.0 g/kg/day for 13 weeks in order to require the no-toxic dose. RESULTS: 1. Soft stool and diarrhea were observed at the 0.50 g/kg group and above, and vomiting was observed at the 1.0 g/kg group and above. Increased water consumption was observed at the 6.25 g/kg group. No deaths occurred at all groups. 2. In urinalysis, increased urine volume was observed at the 6.25 g/kg group. 3. Blood chemistry showed decreased BUN at the 6.25 g/kg group. 4. There were no drug-related changes in body weight, food consumption, ophthalmological examination, electrocardiography, hematology and pathology. 5. At the end of the recovery period, all these changes observed at the end of the administration period were disappeared. Based on these results, it was considered that the no-toxic dose of lactitol is 0.25 g/kg/day.

Administration, Oral↗
PubMed1994

Mechanistic analysis of renal protection by angiotensin converting enzyme inhibitor in Dahl salt-sensitive rats.

OBJECTIVE: To investigate whether and how renin-angiotensin inhibition attenuates renal injury seen in salt-induced hypertension in Dahl salt-sensitive (Dahl-S) rats. METHODS: Dahl-S rats fed a high-salt (4% sodium chloride) diet for 6 weeks were treated with the angiotensin converting enzyme (ACE) inhibitor alacepril or the angiotensin receptor antagonist losartan for 4 weeks. Functional and morphological alterations in the kidney were investigated. RESULTS: Alacepril decreased systolic blood pressure (SBP). This SBP reduction was associated with the attenuation of cardiac and aortic wall hypertrophy and that of proteinuria and urinary N-acetyl-beta-D-glucosaminidase excretion. Kidney injuries, e.g. glomerular, arterial and tubular damage, were improved with alacepril treatment. Losartan decreased SBP to the same extent as alacepril, but neither renal function nor morphological structure was improved as was the case with alacepril. The response of the renal eicosanoid system to alacepril was inadequate, but cyclic GMP excretion, an indicator of nitric oxide formation, was significantly enhanced and lipid peroxidation in the kidney was decreased. CONCLUSIONS: The beneficial effects of ACE inhibition on the renal injury in Dahl-S rats outrange those induced by the receptor antagonism. This might be due to multiple factors including an increased vasodepressor eicosanoid system, enhanced nitric oxide formation and possible inhibition of oxygen radical generation in the injured renal tissues.

Angiotensin-Converting Enzyme Inhibitors↗
PubMed1994

[A case of advanced gastric cancer with liver metastasis disappeared by oral administration of 5'-deoxy-5-fluorouridine after absolute non-curative operation].

A 64-year-old male was found to have a gastric cancer (Borrmann type 3) on the posterior wall of the stomach. Histological diagnosis was poorly differentiated adenocarcinoma. Ultrasonography revealed a hypoechoic lesion (18 x 14 mm) in the anteroinferior segment (S5) of the liver. The biopsy specimens from those lesions proved to be poorly differentiated adenocarcinoma. Total gastrectomy and distal pancreatosplenectomy were performed. During the operation, other liver metastases were found in the anterosuperior segment (S8) and inferolateral segment (S3). 5'-deoxy-5-fluorouridine (800 mg/day) was administered per os from the 33 rd postoperative day. The hypoechoic lesion of S5 in liver disappeared in the 17th postoperative month. The man is healthy with no evidence of recurrence 47 months after the operation. This case is very important because of the long-term CR.

Adenocarcinoma↗
PubMed1994

Light and electron microscopic radioautographic study on the incorporation of 3H-thymidine into the lung by means of a new nebulizer.

The aim of this study was to investigate whether the water particles produced by a nebulizer can reach the alveoli of the lung when they are inhaled with the air. For the purpose of demonstrating water particle incorporation into the lung, light and electron microscopic radioautography with tritiated thymidine was employed. Tritiated thymidine was dissolved in distilled water and nebulized to fine water particles with a new type nebulizer named Shinki, then inhaled by mice. The lungs were taken out and processed by either rapid-freezing and freeze-substitution for dry-mounting radioautography or conventional chemical fixation for wet-mounting radioautography. By wet-mounting radioautography, silver grains were observed in the nuclei of a few alveolar type 2 cells and interstitial cells, demonstrating DNA synthesis. By dry-mounting radioautography numerous silver grains were located diffusely in all the epithelial and interstitial cells, demonstrating diffuse localization of soluble radioisotope-labeled compounds. These results show that water particles produced by the Shinki nebulizer can directly reach the alveoli and thus, better therapeutic effect can be acquired with the Shinki nebulizer than with the usual nebulizer. Further, the nebulizer can be used for the radioautographic studies of the lung in demonstrating incorporation of some drugs into alveolar tissues.

Animals↗
PubMed1994

Genes involved in the dedifferentiation of plant cells.

Since the initial process of culturing tobacco mesophyll protoplasts can be considered as a model system of dedifferentiation of higher plants, the mode of expression of genes induced by auxin, a key factor in inducing dedifferentiation, has been analyzed during the regaining of meristematic activity of quiescent and differentiated tobacco mesophyll. By differential screening we have isolated three auxin-regulated genes, which we named parA, parB and parC. parA and parC, which belong to the same gene family, were supposed to play a role in transcriptional regulation upon induction by auxin, while parB encoded glutathione S-transferase. Although it was supposed that the expression of these par genes should play a pivotal role in regaining the meristematic activity of the differentiated tobacco mesophyll cells, a possibility that other less abundantly expressing genes would have been neglected in these studies has not been excluded. On the other hand, the search for genes which would be involved in maintaining cell division activity in the dedifferentiated plant cells allowed us to isolate a few genes. One of these genes, designated arcA, belonged to a beta subunit-like protein of heterotrimeric G proteins. The significance of the involvement of this gene product in maintaining the meristematic activity of plant cells cultured in vitro has been discussed.

Cell Differentiation↗
PubMed1994

Age-related alterations of proteoglycan in mouse tracheal cartilage matrix: an electron histochemical analysis with the cationic dye of polyethyleneimine.

Age-related alterations of the proteoglycans (PGs) contents and size were observed in the tracheal cartilage matrix of ddY mice at various ages with the cationic dye of polyethyleneimine (PEI). In young animals (aged 1 day to 1 month after birth), the PGs content and size increased, resulting in the formation of the territorial matrix (TM) and the decrease of the space between the PEI aggregates and the collagen fibers. In 1 month old mice, the TM could be distinguished from the interterritorial matrix (ITM). In adult mice (from 2 to 6 months), the PGs content and size kept stable, but the rearrangement of the PGs and collagen fibers was observed. From 6 months of age on, the decrease of PGs content and size occurred mainly in the ITM. The developing stopping sign was recognized in 1 month and the senile sign from 6 months in the ddY mice as for the complete formation of the TM in 1 month and the decrease of the PGs content and size from 6 months. These results demonstrated the age-related alteration of PGs in the ultrastructural level first and could contribute to the aging study of the tracheal cartilage.

Aging↗
PubMed1994

Radioautographic study of benidipine hydrochloride. Localization in the mesenteric artery of spontaneously hypertensive rat.

Microradioautographic study of [3H]-benidipine hydrochloride (KW-3049, CAS 91599-74-5) and nitrendipine (CAS 39562-70-4) localization was performed in the supramesenteric arteries of the spontaneously hypertensive rats (SHR) at 10 and 30 min after intravenous administration or in vitro incubation. The distribution of silver grains was observed by light and electron microscopy and the number of grains was counted in the tunicae intima, media and adventitia of the arteries. Areas of the correlated part of the artery were measured and the grain count per area was calculated. The grain count of [3H]-benidipine hydrochloride showed almost uniform distribution in the tunicae intima and media, which was compared at 10 and 30 min after the administration both in the in vivo and in the in vitro studies. The grain density of [3H]-nitrendipine to the tunicae intima and media increased time-dependently. Electron microscopic observation revealed that the fine distribution of [3H]-benidipine hydrochloride was mainly localized on the plasma membrane and on the cytoplasm of the cellular components of the mesenteric artery. The result suggested that the distribution of [3H]-benidipine hydrochloride was related to its pharmacologically active site, i.e. plasma membrane of the tunica intima and the smooth muscle cells of the tunica media.

Animals↗
PubMed1994

[Peritoneal dissemination of cecal cancer disappearing in a patient taking 5'-DFUR for 3 years].

A 51-year-old woman presenting with a left ovarian tumor underwent left oophorectomy and hysterectomy with intraperitoneal administration of cisplatin, cyclophosphamide, and 5-fluorouracil on December 13, 1989. Microscopically the tumor was well-differentiated adenocarcinoma. Postoperatively cecal cancer was detected. Laparotomy revealed multiple peritoneal dissemination on March 23, 1990, when palliative ileocecal resection was performed. Microscopy revealed several of the nodules to be adenocarcinoma and the cecal lesion well differentiated adenocarcinoma. The patient was then placed on 5'-deoxy-5-fluorouridine (800 subsequently 600 mg/day). In November, 1991, bright red blood per rectum led to the discovery of rectal cancer. On January 26, 1993, laparotomy revealed rectal tumor invading the bladder but the metastatic nodules on the peritoneum had disappeared. Thus pelvic exenteration was performed. The tumor was well-differentiated adenocarcinoma. The ovarian lesion may have been a metastasis from the cecal cancer, a so-called Krukenberg tumor. The rectal cancer was most likely to be asynchronous cancer. 5'-DFUR seemed effective in remitting the peritoneal dissemination.

Adenocarcinoma↗
PubMed1994

Pharmacokinetics and postmortem changes of cimetidine in body tissues.

Cimetidine concentrations in body tissues were pharmacokinetically examined, together with their postmortem changes, to assess the toxicological effect of this drug from the aspect of forensic medicine. For the pharmacokinetic study, rats were pithed at 0.5, 1.0, 2.0, 4.0 or 8.0 hours after an intravenous injection of the drug (33.3 mg/kg), and their tissue samples were immediately collected to be analyzed by high-performance liquid chromatography (HPLC). The concentrations of cimetidine in the blood and muscle decreased below the lower detection limit at 8 hours after the injection, while the drug could still be detected in the brain, lung, liver, kidney, spleen and adipose tissue. The ratios of the cimetidine concentrations in the muscle, lung and spleen to that in the blood were nearly constant until 4 hours after the injection, while those in the liver, kidney and adipose tissue increased as time elapsed. Postmortem changes of cimetidine concentrations were examined using the tissue samples collected 0, 1 and 2 days after death. There was no statistical significance in the postmortem changes in cimetidine concentrations in the body tissues over the two-day period of examination. The experimental results indicated that the muscle, lung and spleen together can serve as samples for toxicological analysis to evaluate the effect of cimetidine, regardless of various postmortem factors, including the availability of blood.

Animals↗
PubMed1993

Isolation of cDNA of an auxin-regulated gene encoding a G protein beta subunit-like protein from tobacco BY-2 cells.

The addition of 2,4-dichlorophenoxyacetic acid to tobacco BY-2 cells that had been cultured in modified Linsmaier and Skoog medium deprived of auxin for 3 days induced cell division, whereas without 2,4-dichlorophenoxy-acetic acid application, no such induction of cell division was seen. When differential cDNA screening for auxin was done at 4 hr after the addition of 2,4-dichlorophenoxyacetic acid, the cDNA of an auxin-responsive gene designated arcA was isolated. The predicted gene product of arcA is a polypeptide with a M(r) of 35,825. arcA, thus, is a plant hormone-regulated gene that encodes a protein structurally related to the beta subunit of a guanine nucleotide-binding regulatory protein, which is composed of seven repetitive segments of Trp-Asp 40-aa repeats. The possibility that arcA gene products induce cell division is discussed.

2,4-Dichlorophenoxyacetic Acid↗
PubMed1993

Escherichia coli aromatic amino acid aminotransferase: characterization and comparison with aspartate aminotransferase.

Aromatic amino acid aminotransferase (ArAT) from Escherichia coli was overexpressed in E. coli cells, purified, and characterized. The enzyme was similar to aspartate aminotransferase (AspAT) of E. coli in many aspects, such as gross protein structure and spectroscopic properties. The reactions of pyridoxal 5'-phosphate-form ArAT with amino acids and pyridoxamine 5'-phosphate-form ArAT with oxo acids were investigated using stopped-flow spectrophotometric techniques. The kinetic parameters for these "half" reactions could excellently explain the ArAT-catalyzed overall transamination reactions at pH 8.0. Reactions of ArAT with aspartate and tryptophan which had been deuterated at position 2 showed isotope effects of 2.5 and 6.0 in the kcat values of the half-reactions, showing that the proton-transfer step is at least partially rate-limiting for these reactions. ArAT and AspAT showed overlapping substrate specificity. Both ArAT and AspAT were active toward dicarboxylic substrates. ArAT showed, however, 10(3)-fold higher activity toward aromatic substrates than AspAT. This high activity toward aromatic substrates was in part ascribed to the active site hydrophobicity of ArAT, which was suggested to be about 1.4 times as large as that of AspAT. In addition to dicarboxylic substrate analogs, aromatic substrate analogs such as carboxylic acids, 2-methyl amino acids, and 3-hydroxy amino acids caused characteristic changes in the absorption spectra of ArAT, while these aromatic analogs did not significantly change the spectra of AspAT. In particular, the erythro-3-hydroxy analogs of phenylalanine and aspartate caused a prominent absorption of ArAT at around 500 nm, which is generally ascribed to the accumulation of quinonoid intermediates. The threo forms of these 3-hydroxy analogs acted as substrates for ArAT. The erythro and threo forms of 3-hydroxyaspartate reacted with AspAT similarly as they reacted with ArAT; however, both forms of 3-phenylserine were poor substrates for AspAT, although phenylalanine was a fairly good substrate for AspAT. The observations on the two erythro-3-hydroxy amino acids show the similar orientation of these analogs in the active site of ArAT, probably through a hydrogen-bonding network involving the hydroxy groups of the analogs and Tyr70, and suggest that the aromatic binding pocket is near or even overlaps the side-chain-carboxylate-binding site for dicarboxylic substrates.

Amino Acid Sequence↗
PubMed1993

Molecular cloning of a POU domain-containing factor involved in the regulation of the Bombyx sericin-1 gene.

The POU domain is a highly conserved region found in a number of transcription factors and products of developmental control genes. We report here the isolation and characterization of a POU domain-containing cDNA (POU-M1) from the middle silk gland of Bombyx mori. It encodes a protein with a POU domain identical to that of the Drosophila Cf1-a protein. By mobility shift and nuclease protection assays, the POU-M1 protein and the putative silk gland factor 3 (SGF-3) were found to interact in an indistinguishable manner with the SC region (positions -204 to -183) of the sericin-1 gene, a key cis-acting element involved in the regulation of the gene through the interaction with SGF-3. Antibodies raised against the synthetic oligopeptide corresponding to the COOH-terminal region of the putative POU-M1 sequence reacted specifically to both the POU-M1 protein and SGF-3. Northern blot hybridization and Western blotting revealed that POU-M1 expression is regulated both temporally and spatially during silk gland development. We conclude that the POU-M1 protein is SGF-3 and propose that the differential expression of the POU-M1 gene is probably involved in the transcriptional regulation of the silk protein genes.

Amino Acid Sequence↗
PubMed1993

Simultaneous determination of paraquat and diquat in human tissues by high-performance liquid chromatography.

A simple, sensitive, reliable, and economical method for simultaneous determination of paraquat dichloride and diquat dibromide in human biological materials has been developed, using high-performance liquid chromatography. The drugs were extracted from the sample with a Sep-Pak C18 cartridge and applied to a chromatograph with the internal standard, L-tyrosine. Paraquat and diquat were clearly separated on the octadecylsilica column with a mobile phase of 0.5% potassium bromide in 5% methanol solution, containing triethylamine (1 ml/l). The pH of the mobile phase was adjusted to 3-4 with 1.3 M phosphoric acid. Two ultraviolet wavelengths were selected, 256 nm for paraquat as well as the internal standard, and 310 nm for diquat. The calibration curves were linear in the concentration range 0.1-10 micrograms/g, and the lower limit of detection was 0.05 microgram/g. We used this method to examine the concentrations of paraquat and diquat in tissues of an individual at autopsy.

Adult↗