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Biomedical subjects

Y Kikuchi

Publications and source records attributed to Y Kikuchi.

At least 415 records · Page 23Linked to original sources
PubMed1994

The MCS1/SSD1/SRK1/SSL1 gene is involved in stable maintenance of the chromosome in yeast.

A temperature-sensitive (ts) mutant of Saccharomyces cerevisiae was isolated in which mini-chromosomes were unstable at high temperature. The MCS1 gene (Mini-Chromosome Stability 1) was cloned by the ability of complementing the temperature sensitivity, and was found to be identical to SSD1/SRK1/SSL1. When MCS1/SSD1 was disrupted in a certain wild-type (wt) strain, mini-chromosomes were unstable, even at 30 degrees C, indicating that the gene is involved in chromosome stability. The Mcs1/Ssd1 protein was detected as a 170-kDa protein by immuno-blotting analysis and this 170-kDa protein could not be detected in the ts mutant and certain wt strains. Our results are consistent with the genetic data that there are two polymorphic forms of the gene, SSD1-v and ssd1-d [Sutton et al., Mol. Cell. Biol. 11 (1991) 2133-2148]. Furthermore, genetic backgrounds other than MCS1/SSD1 caused strain-specific phenotype. The protein, precipitated by specific antibodies, was phosphorylated.

Chromosome Deletion↗
PubMed1994

Chemosensitivity and perception of dyspnea in patients with a history of near-fatal asthma.

BACKGROUND: Many deaths from attacks of asthma may be preventable. However, the difficulty in preventing fatal attacks is that not all the pathophysiologic risk factors have been identified. METHODS: To examine whether dyspnea and chemosensitivity to hypoxia and hypercapnia are factors in fatal asthma attacks, we studied 11 patients with asthma who had had near-fatal attacks, 11 patients with asthma who had not had near-fatal attacks, and 16 normal subjects. Their respiratory responses to hypoxia and hypercapnia, determined by the standard rebreathing technique while the patients were in remission, were assessed in terms of the slopes of ventilation and airway occlusion pressure as a function of the percentage of arterial oxygen saturation and end-tidal carbon dioxide tension, respectively. The perception of dyspnea was scored on the Borg scale during breathing through inspiratory resistances ranging from 0 to 30.9 cm of water per liter per second. RESULTS: The mean (+/- SD) hypoxic ventilatory response (0.14 +/- 0.12 liter per minute per percent of arterial oxygen saturation) and airway occlusion pressure (0.05 +/- 0.05 cm of water per percent of arterial oxygen saturation) were significantly lower in the patients with near-fatal asthma than in the normal subjects (0.60 +/- 0.35, P < 0.001, and 0.16 +/- 0.08, P < 0.001, respectively) and the patients with asthma who had not had near-fatal attacks (0.46 +/- 0.29, P = 0.003, and 0.15 +/- 0.09, P = 0.004). The Borg score was also significantly lower in the patients with near-fatal asthma than in the normal subjects, and their lower hypoxic response was coupled with a blunted perception of dyspnea. CONCLUSIONS: Reduced chemosensitivity to hypoxia and blunted perception of dyspnea may predispose patients to fatal asthma attacks.

Adult↗
PubMed1994

Biochemical and functional characterization of soluble form of IL-5 receptor alpha (sIL-5R alpha). Development of ELISA system for detection of sIL-5R alpha.

Interleukin-5 (IL-5) mediates pleiotropic functions in various types of cells through its specific receptor (IL-5R) which is composed of two distinct subunits, alpha and beta. In mice, the alpha subunit (IL-5R alpha) specifically binds IL-5 with low affinity. The beta subunit (IL-5R beta) does not bind IL-5 by itself, but constructs the high affinity receptor with IL-5R alpha. We have isolated cDNA encoding the soluble form of IL-5R alpha (sIL-5R alpha). To elucidate the biochemical and functional properties of sIL-5R alpha, we developed an expression system for sIL-5R alpha cDNA in insect cell line Sf21 using baculovirus expression vector and obtained conditioned medium containing large quantities of mouse sIL-5R alpha. Mouse sIL-5R alpha was purified from the conditioned medium by using anti-IL-5R alpha mAb-coupled beads. Immunoaffinity-purified sIL-5R alpha with an approximate molecular mass of 42 kDa inhibited the binding of 125I-labeled IL-5 to IL-5R. By using purified sIL-5R alpha, we prepared rabbit anti-sIL-5R alpha antibody and developed a sandwich ELISA for detection of sIL-5R alpha. Significant amounts of sIL-5R alpha were detected in sera and ascitic fluids of mice bearing tumors (BCL1 and MOPC104E) that responded to IL-5 for DNA synthesis, but not in sera of normal mice. Interestingly, elevated levels of serum sIL-5R alpha were observed in NZB and NZW mice. The sIL-5R alpha may, therefore, have an immunoregulatory role in vivo.

Animals↗
PubMed1994

Modified cell-flow microchannels in a single-crystal silicon substrate and flow behavior of blood cells.

Previously reported cell-flow microchannels in a single-crystal silicon substrate (Microvasc. Res. 44, 226-240, 1992) have been modified, and flow behavior of blood cells is described using flow rate-time curves and video pictures. The principal structure (2600 identically sized channels in parallel) was retained to give the same simple quantitative measure of the total flow rate for blood cell suspensions under constant suction. Level areas (terraces) were placed at the entrance and exit sides of the parallel channels level with the channel depth (4.5 microns) so that blood cells just entering into and flowing out of the channels could be more clearly observed under reflecting illumination. Three lengths (10, 20, and 100 microns) of channel were used each with a terrace width of 30 microns. In agreement with calculated values, the resistance to flow at the terrace portion was shown to be nearly equal to that per 10 microns of the channel portion. Clearer pictures were obtained of channel blocking by activated leukocytes and platelet aggregates after addition of each stimulant. Erythrocyte aggregates showed easy transit even through the 100-microns-long channels and through narrow spaces, including gaps probably narrower than 2 microns, which were formed between plugging leukocytes at the terrace portion.

Blood Flow Velocity↗
PubMed1994

Variation in red blood cell deformability and possible consequences for oxygen transport to tissue.

Red blood cell (RBC)-plasma suspensions with a hematocrit of 10-12% were forced to flow through microchannels (equivalent diameter 6 microns, equivalent length 20 microns, 2600 in parallel) by applying a pressure difference of 20 cm H2O. Transit times for 100 microliters of fresh suspensions from 33 healthy male subjects aged between 24 and 63 ranged from 43.0 to 65.2 sec (mean +/- SD: 49.4 +/- 4.3 sec). Four measurements taken from two subjects at intervals over several weeks gave fluctuations of 45.1, 43.6, 47.6, and 45.9 sec and 52.3, 49.8, 60.1, and 50.0 sec, respectively. RBC deformability thus appeared to vary considerably between different subjects and also showed fluctuations between different days in any given subject. No correlation was obtained between the transit time and subject's age. However, higher whole blood hematocrit values were apparent in those suspensions that had longer transit times. A lower RBC deformability might be caused by dietary factors with resultant increased frequency of occasional stagnation of capillary blood flow, i.e., occasional tissue hypoxia. This would further result in an increased hematocrit.

Adult↗
PubMed1994

Polymerase chain reaction-single strand conformation polymorphism analysis of the p53 gene in paraffin-embedded surgical material from human renal cell carcinomas.

p53 tumour suppressor gene mutations were studied in 118 renal cell carcinomas using paraffin-embedded surgical material. Optimal results were obtained with analysis of exon lengths between 150 and 200 base pairs for polymerase chain reaction. Single strand conformation polymorphism and sequencing analysis revealed only two point mutations (2/118, 2%): one involving codon 135; TGC-->TTC (cysteine-->phenylalanine) and the other codon 175; CGC-->CAC (arginine-->histidine). Both of these cases were classified as granular cell subtype on microscopic observation. The data suggest that the p53 tumour suppressor gene is not related to tumour initiation, promotion, or progression of renal cell carcinomas. However, there is the possibility that granular cell type carcinomas may have a different genetic background from clear cell type renal neoplasms.

Base Sequence↗
PubMed1994

Immunocytochemical survey of the neuroepithelial endocrine system in the respiratory tract of the Tokyo salamander, Hynobius nebulosus tokyoensis TAgo.

The epithelial lining of the respiratory tract of urodeles has been shown to harbor an innervated system of neuroepithelial endocrine (NEE) cells. Even between phylogenetically closely related species, large differences have been reported in the appearance and chemical coding of the NEE system. Although urodeles are well suited for the purpose, none of the prior studies have provided an immunocytochemical survey of the NEE system in all parts of the respiratory tract. In the present study, many bioactive substances and a general marker were immunocytochemically demonstrated in serial sections of the entire respiratory tract of the Tokyo salamander, Hynobius nebulosus tokyoensis, a species in which neuroepithelial bodies (NEBs) were previously characterized at the electron microscopic level. In the current study, serotonin-immunoreactive solitary NEE cells were observed in variable numbers in the larynx, in all parts of the trachea, and in areas of the lungs covered with ciliomucous epithelium. Serotonin-containing NEBs, however, were detected in small cranial areas of the lung only. Solitary NEE cells were seen in the trachea and lungs of H. nebulosus tokyoensis by immunocytochemical staining for somatostatin, calcitonin, calcitonin gene-related peptide, and bombesin, but the number, localization, and appearance of the labeled NEE cells differed considerably. Only calcitonin-like immunoreactivity was also noted in some NEB-like cell clusters in the cranial parts of the lungs. Unlike many other vertebrates, neuron specific enolase was found to be a poor marker for the NEE system in the salamander species used in this investigation. It may be concluded that the NEE system of H. nebulosus tokyoensis contains at least five different bioactive substances.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1994

Preclinical studies of antitumor prostaglandins by using human ovarian cancer cells.

Pleiotropic actions of antitumor prostaglandins (PGs) on tumor cells are reviewed including our preclinical results focused on human ovarian cancer. Regarding inhibition of cell proliferation, antitumor PGs exerts its action as a G1 blocking agent. The cyclopentenone PGs inhibit myc oncogene expression while inhibiting the cell cycle progression and results in apoptotic cell death and growth inhibition. Cyclopentenone PGs inhibit growth of various tumors transplanted to mice or nude mice and show adjuvant effects to cis-diamminedichloroplatinum(II) (CDDP). In order to elucidate a role of antitumor PGs in immune systems, relevance of effects on tumor growth with those on the immune systems are also discussed with our results.

Animals↗
PubMed1994

Effects of continuous intravenous infusion of isosorbide dinitrate on development of tolerance to vasodilating action in human epicardial coronary arteries.

This study was performed to determine the effects of long-term intravenous infusion on the coronary vasodilating actions of continuous intravenous and bolus intracoronary administration of isosorbide dinitrate (ISDN). With quantitative coronary angiography, the coronary diameter and the vasodilating response to intracoronary ISDN (1 mg) at angiographically normal segments were studied before and after intravenous administration of ISDN, 10 to 60 micrograms/min for 1 hour, 2 days, or 5 days. The vasodilating effects of intravenous ISDN were 72% +/- 13%, 65% +/- 21%, and 6% +/- 11% of the response to intracoronary ISDN in the baseline study in each group. Irrespective of the duration of intravenous infusion, subsequent intracoronary ISDN dilated coronary arteries to extent similar to that observed in each baseline study. In conclusion, significant coronary vasodilating effects of intravenous ISDN were observed after a 2-day infusion, whereas tolerance to the vasodilating effects apparently developed within 5 days of infusion. The vasodilating response to bolus intracoronary ISDN was preserved even when the vasodilating effects of intravenous ISDN were no longer present.

Angina Pectoris↗
PubMed1994

Clinical experience of Hokkaido University-PACS and FCR-angiography.

Three years' experience with Hokkaido University-PACS (HU-PACS) is reported. In particular, this paper describes the suitability of FCR-angiography for HU-PACS, which has been in clinical use since March 1991. Image quality of FCR-arteriograms was evaluated in the head-and-face region and the abdominal region independently. The image quality in both regions was excellent. Quality of transferred images to image workstation for HU-PACS with 10:1 data compression was also evaluated, and no appreciable image degradation or loss of information was found in the transferred images. There was no significant difference in the examination time required for one patient in abdominal angiography between conventional angiography and FCR-angiography. In summary, FCR-angiography is suitable for HU-PACS as its image acquisition modality.

Abdomen↗
PubMed1994

Preproenkephalin gene expression in the rat cerebral cortex during chronic tracheal stenosis.

To evaluate whether the endogenous opioid system is activated in the higher brain centre while a chronic resistance to airflow, we examined changes of mRNAs for preproenkephalin (PPE)-A, which is a precursor of enkephalin, and for 70 kD heat shock protein (HSP70) in the cerebral cortex of rat brain during chronic tracheal stenosis. Northern blot revealed that PPE-A mRNA was induced at 3 days of airway stenosis. In situ hybridization revealed that PPE-A mRNA was gradually induced in frontal cortex. The significant induction of PPE-A mRNA was observed at 3 and 7 days. However, no significant induction of HSP70 gene was observed. These results suggest that the endogenous opioid system may be at work as an important compensatory mechanism to reduce the respiratory sensation during chronic respiratory stress.

Animals↗
PubMed1994

Effects of galanin on amylase secretion from dispersed rat pancreatic acini.

Dispersed rat pancreatic acini were used to determine the effect of galanin on the exocrine pancreas and on basal and secretagogue-stimulated amylase secretion. Basal amylase secretion and amylase release stimulated by cholecystokinin octapeptide, bombesin, 12-o-tetradecanoyl-phorbol-13-acetate (TPA), secretin, and vasoactive intestinal peptide were not affected by galanin in doses ranging from 10(-12) to 10(-6) M. Galanin, however, significantly inhibited the amylase release stimulated by sub- and supramaximal doses of carbachol. A time course study showed that the inhibition by galanin occurred during the sustained phase of carbachol-stimulated amylase secretion. The inhibitory action of galanin disappeared in acini obtained from animals pretreated with pertussis toxin (PTX). These results suggest that galanin inhibits carbachol-stimulated amylase secretion through a mechanism related to a PTX-sensitive G protein.

Amylases↗
PubMed1994

Role of nitric oxide in the pancreatic blood flow response to caerulein.

To clarify the role of nitric oxide (NO) in the pancreas, blood flow in the rat pancreas (pancreatic blood flow: PBF) was investigated by the hydrogen clearance technique using a specific NO synthase inhibitor, N omega-nitro-L-arginine (L-NNA). Continuous infusion of caerulein at doses of 5 and 20 micrograms/kg/h caused a significant increase in PBF in the early phase of caerulein infusion. The caerulein-induced increase in PBF was not affected significantly by atropine sulfate (100 micrograms/kg), nor by phenoxybenzamine (5 mg/kg) plus propranolol (50 micrograms/kg). Administration of L-NNA (0.5, 5, or 30 mg/kg) did not affect the basal PBF, but at 5 mg/kg it inhibited completely the caerulein-induced increase in PBF. The inhibitory action of L-NNA was reversed by a large dose of L-arginine (100 mg/kg bolus, i.v., followed by a continuous infusion at 400 mg/kg/h), but not by its enantiomer D-arginine. These results strongly suggest that NO has a mediator role in the early phase vascular response of the pancreas to superphysiologic doses of caerulein.

Amino Acid Oxidoreductases↗
PubMed1994

Isolation of genes differentially expressed between the Yoshida sarcoma and long-survival Yoshida sarcoma variants: origin of Yoshida sarcoma revisited.

The Yoshida sarcoma (YS) is characterized by growth as "free cells" in ascites. Long-survival Yoshida sarcoma (LY) variants, which develop after transplantation of YS into immunologically conditioned Donryu rats, in contrast, form "islands" in ascites. A representational difference analysis (RDA) approach was adopted to isolate genes differentially expressed between YS and LY variants to elucidate the molecular mechanism of their development. Fifteen clones presenting differences in expression were characterized. Nine genes (those encoding for the high-affinity IgE receptor gamma chain, pJG116 repetitive sequence, non neuronal enolase, proteasome subunit RC1, cytotoxic T lymphocyte-associated gene transcript CTLA-1, interleukin-2 receptor gamma chain, and three unknown sequences) expressed mRNA in YS, but showed lower or no expression of mRNA in LYs. The mRNAs of the other six genes (those encoding for cytokeratin 8, cytokeratin18 (Endo B), TIMP2 and three unknown sequences) were not found in YS, but were present in LYs. Interestingly, CTLA-1 is a non-epithelial (hematopoietic) cell-specific gene in terms of transcription, while cytokeratin 8 and cytokeratin 18 are both epithelium-specific genes. Immunohistochemically, YS expressed T-cell specific antigens CD2 and CD3, and T cell receptor beta and gamma chain genes were rearranged in YS, but not in LYs. Moreover, using restriction fragment length polymorphism probes, we found that LYs exhibited different cell lineage from YS. Thus, our present findings, unexpectedly, raise fundamental questions concerning the cellular origins of YS and LY variants rather than pointing to any specific mechanism to explain the LY phenomenon.

Animals↗
PubMed1994

Direct imaging in a water layer of human chromosome fibres composed of nucleosomes and their higher-order structures by laser-plasma X-ray contact microscopy.

X-ray contact microscopy with a 300-ps-duration laser-plasma X-ray source has been used to image hydrated human chromosomes. Clearly imaged are individual nucleosomes and their higher-order particles (superbeads), elementary chromatin fibrils. c. 30 nm in diameter and their higher-order fibres of various sizes up to c. 120 nm in diameter. The results demonstrate that X-ray microscopy is now capable of opening a new path of investigation into the detailed structures of hydrated chromosome fibres in their natural state.

Cells, Cultured↗
PubMed1994

Fluctuations of the sympathetic nerve discharges in animals without natural heartbeat.

To assess the effect of artificial circulation on the circulatory regulatory system, rhythmical fluctuations in the cardiovascular system and autonomic nerve discharges were analyzed in animal experiments using adult mongrel dogs. Two pneumatically actuated ventricular assist devices were implanted as total biventricular bypass (BVB) under general anesthesia; then, the natural heart was electrically fibrillated to constitute the BVB part of the complete prosthetic circulation model. All hemodynamic data and sympathetic nerve discharges were recorded and calculated in the computer system by the use of spectral analyzing methods. In the power spectrum of the arterial blood pressure waveform during prosthetic circulation, the Mayer wave peak was significantly decreased though the respiratory wave peak was not significantly changed. However, the Mayer wave peak in sympathetic nerve discharges was significantly increased though the respiratory wave peak was not significantly changed. These results suggest that rhythmical fluctuations are a necessary response of the circulatory regulatory system including the autonomic nervous system. However, sympathetic nerve discharges respond to the artificial heart without rhythmical fluctuations.

Animals↗
PubMed1994

Nucleotide sequence and functional analysis of the meta-cleavage pathway involved in biphenyl and polychlorinated biphenyl degradation in Pseudomonas sp. strain KKS102.

Pseudomonas sp. strain KKS102 is able to degrade biphenyl and polychlorinated biphenyls via the meta-cleavage pathway. We sequenced the upstream region of the bphA1A2A3BCD (open reading frame 1 [ORF1]) A4 and found four ORFs in this region. As the deduced amino acid sequences of the first, second, and third ORFs are homologous to the meta-cleavage enzymes from Pseudomonas sp. strain CF600 (V. Shingler, J. Powlowski, and U. Marklund, J. Bacteriol. 174:711-724, 1992), these ORFs have been named bphE, bphG, and bphF, respectively. The fourth ORF (ORF4) showed homology with ORF3 from Pseudomonas pseudoalcaligenes KF707 (K. Taira, J. Hirose, S. Hayashida, and K. Furukawa, J. Biol. Chem. 267:4844-4853, 1992), whose function is unknown. The functions of meta-cleavage enzymes (BphE, BphG, and BphF) were analyzed by using crude extracts of Escherichia coli which expressed the encoding genes. The results showed that bphE, bphG, and bphF encode 2-hydroxypenta-2,4-dienoate hydratase, acetaldehyde dehydrogenase (acylating), and 4-hydroxy-2-oxovalerate aldolase, respectively. The biphenyl and polychlorinated biphenyl degradation pathway of KKS102 is encoded by 12 genes in the order bphEGF (ORF4)A1A2A3BCD (ORF1)A4. The functions of ORF1 and ORF4 are unknown. The features of this bph gene cluster are discussed.

Aldehyde Oxidoreductases↗