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Biomedical subjects

S Kato

Publications and source records attributed to S Kato.

At least 901 records · Page 50Linked to original sources

In-tube cDNA cloning method using a biotinylated RNA probe.

We report a simple, practical method for isolating a particular cDNA from a single-stranded (ss) cDNA library in a tube without using a radioisotope. The method consists of three steps: (i) the capture of a target cDNA by a biotinylated RNA via intermolecular hybrid formation, (ii) the binding of the cDNA-RNA hybrid to an avidin-coated gel, (iii) the recovery of the target cDNA by degradation of the RNA under mild alkaline conditions. The effectiveness of the method was examined by isolating a clone carrying a metapyrocatechase gene from a model library. This model experiment achieved an enrichment of 4800-fold. This method was applied to cloning of a cDNA encoding interleukin 8 (IL-8) from a cDNA library prepared from phorbol myristate acetate-treated U937 cells. Using an RNA probe prepared from a truncated cDNA encoding IL-8, the corresponding full-length cDNA clones were successfully obtained.

Biotin↗

Positive and negative regulation of retinoid X receptor gene expression by thyroid hormone in the rat. Transcriptional and post-transcriptional controls by thyroid hormone.

The 9-cis-retinoic acid receptors (RXRs), belonging to the members of the steroid/thyroid hormone receptor superfamily, act as auxiliary proteins, heterodimerizing with other nuclear receptors such as retinoic acid receptors (RARs), vitamin D receptor, thyroid hormone receptors, and peroxisome-proliferator activated receptor, thereby transactivating target genes in a ligand-dependent manner. We have previously reported that in the rat, thyroid hormone (TH) positively and negatively regulates the hepatic mRNA levels of RXR beta and RXR gamma, respectively. In the present study, we have tried to elucidate the level at which TH regulates the gene expression of RXR beta and RXR gamma in the rat. A RNA synthesis inhibitor (actinomycin D), but not a protein synthesis inhibitor (cycloheximide), blocked the induction of RXR beta mRNA by TH. On the other hand, none of these drugs inhibited the decrease of RXR gamma mRNA levels caused by TH. Nuclear run-on assays showed that the transcription rate of the RXR beta gene was positively regulated by TH, whereas the transcription of RXR gamma gene was not controlled by TH. Taken together, these results indicate that the gene expression of RXR beta is positively regulated by TH at transcriptional level, while the negative regulation of the RXR gamma gene expression by TH may occur at a post-transcriptional level in intact rat. Thus, the RXR-mediated signal transductions may be modulated in part through TH control of the levels of RXR beta and RXR gamma.

Animals↗

Alteration of asparagine-linked glycosylation in serum transferrin of patients with hepatocellular carcinoma.

The asparagine-linked sugar chains in serum transferrin purified from patients with hepatocellular carcinoma (n = 13), healthy individuals (n = 5) and patients with liver cirrhosis (n = 6) were compared. Sugar chains released with N-glycanase from desialylated and pepsin-digested transferrin were derivatized by reductive pyridylamination. Analysis of the sugar chains by high performance liquid chromatography in combination with exoglycosidase digestion revealed an increase of a biantennary complex-type sugar chain with a fucosylated trimannosyl core; Gal beta 1-4GlcNAc beta 1-2Man alpha 1-6(Gal beta 1-4GlcNAc beta 1-2Man alpha 1-3) Man beta 1-4GlcNAc beta 1-4(Fuc alpha 1-6)GlcNAc in 7 of 13 cancer patients and an increase of a sugar chain with a fucosylated trimannosyl core and bisecting N-acetylglucosamine; Gal beta 1-4GlcNAc beta 1-2Man alpha 1-6(GlcNAc beta 1-4) (Gal beta 1-4GlcNAc beta 1-2Man alpha 1-3)Man beta 1-4GlcNAc beta 1-4(Fuc alpha 1-6)GlcNAc in one of the 13 cancer patients. Further, the fucosylated alteration of the sugar chain was detected also in alpha 1-antitrypsin, hemopexin, alpha 1-acid glycoprotein and alpha 2-HS glycoprotein from one of the patients with increased fucosylated transferrin.

Asparagine↗

Identification of anti-agglutinin for spermatozoa in epididymal boar plasma.

The present report identifies epididymal boar anti-agglutinin and examines its effect on sperm motility. Boar spermatozoa from the cauda epididymidis were washed and incubated in modified Krebs-Ringer bicarbonate at 37 degrees C (5% CO2 in air). In the samples washed three or five times and then incubated for 3-5 h, higher rates (72-79%) of spermatozoa were associated with one another at the acrosomal region, mainly in groups of 2-5 cells (head-to-head agglutination), and many cells exhibited intensively flagellant and/or circular types of movement but rarely progressive motility. The addition of epididymal plasma or 25 kDa protein purified from it markedly inhibited the occurrence of head-to-head agglutination in washed spermatozoa, whereas heat treatment and subsequent removal of insoluble materials reduced the anti-agglutination activity of epididymal plasma. The percentages of progressively motile cells in the samples incubated with epididymal plasma or 25 kDa epididymal protein rose coincident with the reduction of sperm agglutination. These findings demonstrate that the 25 kDa epididymal protein is an anti-agglutinin for the cauda spermatozoa and that it effectively functions to maintain progressive motility of the cells in vitro.

Animals↗

Hepatocyte growth factor immobilized onto culture substrates through heparin and matrigel enhances DNA synthesis in primary rat hepatocytes.

Newly prepared phosphatidylethanolamine (PE) conjugates of glycosaminoglycans can be immobilized to solid phase through hydrophobic interaction. Primary rat hepatocytes were cultured on type I collagen or laminin substrates containing heparin-PE and assayed for DNA synthesis initiated by hepatocyte growth factor (HGF). Hepatocytes responded to HGF that had been added to culture media by active DNA synthesis and pronounced cell spreading, regardless of the presence of heparin-PE on the substrates. The preincubation with HGF, but not with epidermal growth factor, of the substrates containing heparin-PE significantly enhanced DNA synthesis and cell spreading in the absence of these mitogens in culture media. The enhancements were abolished by omitting heparin-PE or washing the HGF-treated substrates with 1 M NaCl, suggesting the immobilization of HGF to substrates through heparin. These phenomena were more evident with laminin substrates than with type I collagen substrates. Chondroitin sulfate-PE only partly substituted for heparin-PE, but Matrigel gave results similar to those of the substrates containing heparin-PE. Both type I collagen and laminin substrates containing heparin-PE bound higher amounts of HGF than the respective control substrates, whereas neither of the substrates containing chondroitin sulfate-PE did. However, the increase in HGF binding to the substrates was not as evident as the increase in DNA synthesis, suggesting that the latter is not simply due to the former but due to a concerted action of the immobilized HGF and heparin. Taken together, these results suggest that HGF can be trapped in extracellular matrix, probably through heparan sulfate in vivo, thereby acting as a mitogen for hepatocytes in cooperation with heparan sulfate.

Animals↗

Ambient pressure stimulates immortalized human aortic endothelial cells to increase DNA synthesis and matrix metalloproteinase 1 (tissue collagenase) production.

In the present study, we investigated the effect of ambient pressure on [3H]-thymidine incorporation and on the production of matrix metalloproteinase 1 (tissue collagenase/proMMP-1) using human aortic endothelial cells immortalized with simian virus 40 (SE-1). Incubation of cells at ambient pressures of 50 and 100 mmHg for 24 h slightly increased [3H]-thymidine incorporation when directly compared with normal culture conditions. The amount of [3H]-thymidine incorporated in SE-1 reached a maximum at 150 mmHg, while a further increase in pressure to 200 mmHg decreased incorporation. The same ambient pressure slightly stimulated human aortic intimal smooth muscle cells (SMC) to increase [3H]-thymidine incorporation but not medial SMC. Immunoblot analysis also showed that ambient pressure, ranging from 50 to 200 mmHg, like 12-O-tetradecanoyl-phorbol-13-acetate stimulated SE-1 to produce proMMP-1, an effect not seen with either intimal or medial SMC. The amount of proMMP-1 produced also reached a maximum level at 150 mmHg. We postulate that human endothelial cells are ambient pressure sensitive and that relatively lower ambient pressures play an important role in the growth of endothelial cells, while higher pressures injure endothelial cells, resulting in the initiation of atherosclerosis. This cell line may prove useful in the investigation of both the physiological and pathological roles of blood pressure on endothelial cell function.

Aorta↗

Demonstration of neurofibrillary tangles and neuropil thread-like structures in spinal cord white matter in parkinsonism-dementia complex on Guam and in Guamanian amyotrophic lateral sclerosis.

This report concerns the demonstration and distribution of neurofibrillary tangles (NFTs), immunoreactive neuropil thread-like structures and dots in the spinal cord gray and white matter of six patients with parkinsonism-dementia complex on Guam and five patients with Guamanian amyotrophic lateral sclerosis (ALS). A monoclonal antibody to Alzheimer NFTs was used. NFTs were found in the spinal cord gray matter and white matter and all patients had immunoreactive neuropil thread-like structures and dots in the gray matter as well as in the white matter. They were particularly numerous in the lateral funiculus of patients with Guamanian ALS.

Amyotrophic Lateral Sclerosis↗

Reduced sensitivity of dihydroxyacetone on ATP-sensitive K+ channels of pancreatic beta cells in GK rats.

In the GK (Goto-Kakizaki) rat, a genetic model of non-insulin-dependent diabetes mellitus, glucose-induced insulin secretion is selectively impaired. In addition, it has been suggested by previous studies that impaired glucose metabolism in beta cells of the GK rat results in insufficient closure of ATP-sensitive K+ channels (KATP channels) and a consequent decrease in depolarization, leading to a decreased insulin release. We have recently reported that the site of disturbed glucose metabolism is probably located in the early stages of glycolysis or in the glycerol phosphate shuttle. In the present study, in order to identify the impaired metabolic step in diabetic beta cells, we have investigated insulin secretory capacity by stimulation with dihydroxyacetone (DHA), which is known to be directly converted to DHA-phosphate and to preferentially enter the glycerol phosphate shuttle. In addition, using the patch-clamp technique, we also have studied the sensitivity of DHA on the KATP channels of beta cells in GK rats. The insulin secretion in response to 5 mmol/l DHA with 2.8 mmol/l glucose was impaired, and DHA sensitivity of the KATP channels was reduced in beta cells of GK rats. From these results, we suggest that the intracellular site responsible for impaired glucose metabolism in pancreatic beta cells of GK rats is located in the glycerol phosphate shuttle.

Adenosine Triphosphate↗

Enhancement effects of BSA and linoleic acid on hybridoma cell growth and antibody production.

The effects of linoleic acid and bovine serum albumin on hybridoma cell growth and antibody production were investigated. In dish cultivation, linoleic acid on its own promoted cell growth when used at concentrations below 50 mg L-1, but strongly inhibited growth at a concentration of 100 mg L-1 on more. However, linoleic acid bound to bovine serum albumin did not inhibit cell growth, even at a concentration as high as 100 mg L-1. Also, linoleic acid did not affect the specific antibody production rate, with or without bovine serum albumin. In order to elucidate the enhancement of antibody production by bovine serum albumin, fractions were prepared by ultrafiltration (98% molecular weight cut-offs, 50,000 and 17,000) and the effects of the fractionation on antibody production were studied in batch cultivation. The high-molecular-weight fraction (> or = 50,000) promoted antibody production whereas the low-molecular-weight fraction (< or = 17,000) inhibited it. In continuous cultivation, the high-molecular-weight fraction was also found to enhance antibody production.

Animals↗

An approach to the hepatic hilus with marked fibrosis: a surgical technique for reconstruction of the bile duct with a benign stricture.

It is very difficult to approach the hepatic hilus safely in patients with common and/or hepatic bile duct strictures in which normal tissue has been replaced by scarred tissue with firm fibrous adhesions. In this report, we describe how eight patients with benign strictures of the bile duct underwent an operation which involved dividing the superior mesenteric and portal veins from the lower margin of the pancreas in a dorsal direction using a finger in a tunneling technique. The common bile duct, which was buried in scar tissue, was then explored, while the common and/or proper hepatic arteries in the hepatoduodenal ligament were confirmed, after transsection at the superior margin of the pancreas. Biliary reconstruction was successfully performed after resection of the constricted bile duct in all the patients, none of whom have experienced recurrence from 6 months to 5 years after the operation.

Aged↗

The strategy to treat disseminated neuroblastoma utilizing bone marrow transplantation: what is the surgeon's role?

The current role of surgery was evaluated in seven consecutive patients with high-risk neuroblastoma (six stage 4 patients and one stage 3, abdominal origin, and all over 12 months of age at diagnosis) treated with new modalities utilizing bone marrow transplantation (BMT). In six of these seven patients, a grossly complete excision of the primary tumor was achieved, and four have survived for 133, 69, 39, and 28 months with no further evidence of disease. The remaining patient with celiac neuroblastoma only underwent a biopsy during a second-look laparotomy after chemoradiotherapy, and thereafter developed local and distant recurrences and ultimately succumbed to the tumor. The timing of surgical intervention varied, either before or after chemotherapy, and did not appear to affect the ultimate survival. Although surgical excision of the primary tumor remains a very high priority in the overall treatment scheme, the most important factor remains the eradication of the tumor by well-planned courses of intensive chemotherapy (e.g., A1 Protocol of the Study Group of Japan). Thus, after having induced complete remission, for consolidation, it is necessary to kill all remaining tumor cells by giving supralethal doses of chemotherapy including total body irradiation (TBI) assisted by BMT.

Adrenal Gland Neoplasms↗

Septum formation of the common hepatic duct associated with an anomalous junction of the pancreaticobiliary ductal system and gallbladder cancer: report of a case.

We herein describe a 48-year-old woman who developed a septum formation of the bile duct combined with an anomalous arrangement of the pancreaticobiliary ductal system in conjunction with gallbladder cancer. A preoperative endoscopic retrograde cholangiogram demonstrated a filling defect in the common hepatic duct which was misdiagnosed to be an elevated lesion. Further exploration revealed a septum formation of the bile duct along with gallbladder cancer. A resection of the dilated bile duct and gallbladder along with a dissection of the regional lymph nodes was performed. A histological evaluation showed the septum to consist of a normal bile duct wall while the tumor in the gallbladder was poorly differentiated adenocarcinoma. The patient made an uneventful recovery. The septum formation was presumed to be congenital in origin.

Adenocarcinoma↗

Cushing's syndrome with cortisol hypersecretion from one of bilateral adrenocortical adenomas: report of a case.

We report herein the case of a 40-year-old man with Cushing's syndrome, diagnosed by clinical manifestations and endocrinological studies, who was found to have bilateral adrenocortical adenomas, one of which hypersecreted cortisol. The Cushing's syndrome was therefore attributed to primary adrenocortical disease, and the right adrenal tumor was resected and histologically diagnosed as a so-called black adenoma. After resection of the right tumor, the left adrenal tumor showed no signs of cortisol hypersecretion for the 23 months of follow-up until the patient died of peritonitis subsequent to the rupture of a duodenal ulcer. The left adrenal tumor was examined at autopsy and found to be a cortical adenoma. These data imply that the adrenal adenomas developed primarily from the adrenal gland itself, and that one of the tumors was well differentiated and secreted excess hormones, while the other remained in cell proliferation without hypersecretion.

Adrenal Gland Neoplasms↗

Endoscopic ethanol injection for treatment of bleeding peptic ulcer.

We treated three children aged 10, 11 and 13 years with actively bleeding ulcers using local endoscopic injection of pure ethanol. Ethanol was injected into several sites around a visible vessel with or without bleeding. Haemostasis following ethanol injection therapy was confirmed by endoscopy performed the day after treatment. No rebleeding was observed. There were no complications related to the procedure. Injection therapy is technically simple and inexpensive. Conclusion Our results suggest that endoscopic ethanol injection is safe and may be the treatment of choice for control of bleeding from peptic ulcers in children.

Adolescent↗

The effects of using a leukocyte removal filter during cold blood cardioplegia.

During myocardial ischemia, neutrophils and platelets exert negative effects on the myocardium. In this study, we used a leukocyte removal filter during cardioplegia, and investigated its effect on myocardial damage during reperfusion by measuring the plasma levels of granulocyte components, platelet components, and cardiac enzymes [creatinine phosphokinase (CK) and creatinine phosphokinase myocardial band (CK-MB)] in 24 patients who underwent cardiopulmonary bypass. The patients were divided into two groups of 12 according to whether or not a filter was placed in the cardioplegic route. Blood samples were drawn directly from the coronary sinus before aortic cross clamping, and at 1, 5, and 15 min after declamping. Group F, which had the filter, showed better cardiac enzyme and lipid peroxidation results than group N, which did not. The results of this study suggest that the application of a filter during cold blood cardioplegia may reduce myocardial damage.

6-Ketoprostaglandin F1 alpha↗

Fluoride profiles in different surfaces of human permanent molar enamels from a naturally fluoridated and a non-fluoridated area.

The surfaces, sites and depths of fluoride profiles of enamel from a naturally fluoridated area (West Hartlepool, WHP, 1.9 parts/10(6) F in drinking water) and a non-fluoridated area (Leeds, LDS, < 0.1 parts/10(6) F in drinking water) were examined in detail. Using a regression curve, y = ax-b, comparisons of fluoride concentrations were made at depths of 1, 3, 5, 10, 20 and 30 microns at occlusal, middle and cervical sites of buccal, lingual, mesial and distal molar enamel surfaces. In the outermost enamels (< 10 mu in depth), fluoride concentrations tended to be lower in the occlusal sites than in the cervical sites. In interior enamels (> 20 microns in depth), the fluoride concentrations of molars from WHP were statistically higher than those from LDS, while few significant differences were found in the outermost layers. In the mesio- and disto-occlusal sites of approximal surfaces, there were some significant differences in the exponential regression coefficients of fluoride profiles between the two groups of molars.

Dental Enamel↗

Circulatory collapse and sudden death in respirator-dependent amyotrophic lateral sclerosis.

Circulatory collapse and sudden death was defined retrospectively as one of the major critical problems among 23 respirator-dependent patients with amyotrophic lateral sclerosis (ALS). Six cases died from sudden cardiac arrest or anoxic encephalopathy following the circulatory collapse. In five among the six cases, sudden death or cardiac arrest occurred during sleep at night. Eight cases had had episodes of marked fluctuation of blood pressure before death, including paroxysmal elevation of blood pressure and heart rate, and successive sudden pressure fall without compensatory tachycardia. The spells of hypotension often occurred during sleep. In addition, the prospective study of diurnal variation of blood pressure, heart rate, plasma norepinephrine and plasma renin activity in nine respirator-dependent ALS patients showed continuous tachycardia and more remarkable nocturnal decrease of blood pressure compared with the control subjects. Plasma norepinephrine levels were constantly higher in the ALS patients particularly in a daytime. These indicate the continuous sympathetic hyperactivity in ALS. We discuss the cause of the circulatory collapse and sudden death in the respirator-dependent ALS patients in terms of the autonomic dysregulatory mechanism or the sympathetic hyperactivity.

Adult↗

Participation of the limbic system and its associated areas in the dementia of amyotrophic lateral sclerosis.

The topographic distribution of degenerative changes in large brain sections from five sporadic amyotrophic lateral sclerosis (ALS) patients with dementia and three without dementia was examined. The dementia characteristics were impaired shifting from one line of thinking to another, perseveration, and emotional disinhibition as well as impairment of cognition, and judgment. Neuropathological examinations showed definite ALS changes in all the patients studied. In addition, the five patients with dementia showed neuronal loss, gliosis, and sponginess of the superficial layers throughout the cerebral cortices, predominantly in the dorsomedial cortex of the temporal tip and the parahippocampal, ambiens, anterior cingulate, rectal, orbital, and insular gyri as well as neuronal loss in the basolateral nucleus of the amygdala, nucleus accumbens, and subiculum of the hippocampus. Ubiquitin-immunoreactive inclusions were present in some neurons in the granular cell layers of the hippocampus. Fibrous gliosis was extensive in the subcortical and deep white matter of the frontotemporal lobes. The affected regions take in the limbic system and its associated areas which are the sources of the psychological problems, including emotional disturbance, experienced by these ALS patients. The psychological problems of ALS need to be investigated in relation to the involvement of the limbic system.

Adult↗