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Biomedical subjects

K Isono

Publications and source records attributed to K Isono.

At least 181 records · Page 10Linked to original sources

Flies in the group Cyclorrhapha use (3S)-3-hydroxyretinal as a unique visual pigment chromophore.

In the class Insecta, retinal and 3-hydroxyretinal are used as chromophores of visual pigments, but the absolute structure of the 3-hydroxyretinal chromophore has yet to be clarified. This study investigates the chirality of 3-hydroxyretinal in the compound eyes of five representative orders of insects. In the orders Odonata, Hemiptera, Neuroptera and Lepidoptera, and suborders Nematocera and Brachycera of the Diptera, only (3R)-3-hydroxyretinal isomers were detected, but dipterans of the suborder Cyclorrhapha (higher flies) had the (3S)-11-cis enantiomer and a mixture of (3R)-all-trans and (3S)-all-trans 3-hydroxyretinal enantiomers; the ratio of the (3R) enantiomer to the sum of both enantiomers of the all-trans isomer was in the range 9-32%. Despite differences in feeding habits, including one species that is a butterfly parasite, all higher flies analysed to date share the same pattern of 3-hydroxyretinal enantiomers, making them a unique group with regard to the nature of the visual pigment chromophore.

Animals↗

Accelerated bone marrow recovery from radiation damage in etoposide-pretreated mice.

PURPOSE: Etoposide inhibits the activity of Topoisomerase II, and possesses radiosensitizing effects on cultured cells. We showed that pretreatment of mice with etoposide 1 day before whole-body irradiation (WBI) had a protective effect against radiation-induced bone marrow death. The number of endogenous colony-forming units (CFU-S) surviving in WBI mice was significantly increased by pretreatment with etoposide. The purpose of the present study is to clarify mechanisms underlying the protective activity of etoposide against radiation-induced bone marrow damage. METHODS AND MATERIALS: The spleen colony-forming cells (CFU-S) of femoral marrow were assayed by the method of Till and McCulloch. The proliferative state of CFU-S was assessed using the technique of Hodgson. The reduction in exogenous CFU-S after hydroxyurea injection was used to determine the fraction size of S-phase in CFU-S. RESULTS: Etoposide pretreatment did not modify the cell survival parameters of exogeneous CFU-S when bone marrow cells were assayed immediately after WBI. Recovery of CFU-S in femoral marrow started earlier in mice receiving etoposide pretreatment than in the radiation alone group. Administration of hydroxyurea revealed that the accelerated recovery of CFU-S was due to an increased population of S phase cells, suggesting etoposide pretreatment shortened generation time of CFU-S. CONCLUSIONS: The mechanism for the radioprotection appears to be accelerated proliferation of survived bone marrow cells rather than a change in cellular sensitivity to radiation.

Animals↗

Detection of minus-strand hepatitis C virus RNA in tumor tissues of hepatocellular carcinoma.

BACKGROUND: Recently, the detection of hepatitis C virus (HCV) antibody has been widely performed for clinical serum testing of HCV infection and can be identified in most hepatocellular carcinoma (HCC) cases in Japan. In the current study, the authors detected not only plus-strand but also minus-strand HCV RNA as a template for RNA replication in hepatocytes in the resected tumors of HCC, and they investigated those significant for HCC. METHODS: The plus-strand and minus-strand HCV RNA were detected using reverse transcription-polymerase chain reaction after the extraction of RNA from the tumor and nontumor tissues of hepatectomized liver, respectively. RESULTS: The detection of HCV RNA in liver tissues from hepatectomized specimens of 20 cases was as follows: in nontumor regions, both of the plus- and minus-strand HCV RNA were detected in 14 cases. Among those cases, plus- and minus-strand HCV RNA were detected in 11 cases, and in another 2 cases, only plus-strand HCV RNA was found in the tumor regions. CONCLUSIONS: These results suggest that HCV RNA is able to replicate in tumor tissues of HCC and may be involved in the development of HCC and also that HCV in remnant hepatocytes may cause the recurrence of HCC as secondary carcinogenesis after hepatectomy.

Adult↗

The yeast nuclear gene MRP-L13 codes for a protein of the large subunit of the mitochondrial ribosome.

The nuclear gene MRP-L13 of Saccharomyces cerevisiae, which codes for the mitochondrial ribosomal protein YmL13, has been cloned and characterized. It is a single-copy gene residing on chromosome XI. Its nucleotide sequence was found to be identical to that of the previously reported ORF YK105. A comparison of the predicted protein sequence of the MRP-L13 gene product and the actual N-terminal amino-acid sequence of the isolated YmL13 protein indicated that the mature protein is preceded by a mitochondrial signal peptide of 86 amino-acid residues, which is the longest among all known mitochondrial ribosomal proteins of S. cerevisiae. No sequence similarity was found to any other ribosomal protein in the current databases. The transcription of MRP-L13 was found to be repressed in the presence of glucose. Its protein product is not strictly essential for mitochondrial functions, but disruption of the gene by insertion of LEU2 noticeably affected cellular growth on non-fermentable carbon sources.

Amino Acid Sequence↗

Phosrestin I undergoes the earliest light-induced phosphorylation by a calcium/calmodulin-dependent protein kinase in Drosophila photoreceptors.

Activation of PI-PLC initiates two independent branches of protein phosphorylation cascades catalyzed by either PKC or Ca2+/calmodulin-dependent protein kinase (CaMK). We find that phosrestin I (PRI), a Drosophila homolog of vertebrate photoreceptor arrestin, undergoes light-induced phosphorylation on a subsecond time scale which is faster than that of any other protein in vivo. We determine that a CaMK activity is responsible for in vitro PRI phosphorylation at Ser366 in the C-terminal tryptic segment, MetLysSer(P)IleGluGlnHisArg, in which Ser(P) represents phosphoserine366. We also demonstrate that Ser366 is the phosphorylation site of PRI in vivo by identifying the molecular species resulting from in-gel tryptic digestion of purified phospho-PRI using HPLC-electrospray ionization tandem quadrupole mass spectroscopy. From these data, we conclude that the CaMK pathway, not the PKC pathway, is responsible for the earliest protein phosphorylation event following activation of PI-PLC in living Drosophila photoreceptors.

Amino Acid Sequence↗

Cloning and characterization of the ribosomal protein genes in the spc operon of a prokaryotic endosymbiont of the pea aphid, Acyrthosiphon kondoi.

To correlate a prokaryotic endosymbiont in the pea aphid, Acyrthosiphon kondoi, with the endosymbionts in related aphid species as well as with free-living bacteria and subcellular organelles, and to study the mode of its gene expression within aphid cells, we have cloned and characterized the genes encoding ribosomal proteins S3, L16, L29, S17, L14, L24, L5, S14, S8, L6, L18, S5, L30, L15 and secretion protein Y (Sec Y) from the S10 and spc ribosomal protein gene operons of this endosymbiont. The organization of these genes is identical to that in Escherichia coli, and their nucleotide sequences are highly similar (87% identity) to the corresponding E. coli genes. They are much less similar to the corresponding chloroplast and mitochondrial genes. The guanine plus cytosine G+C content of the genes of the A. kondoi endosymbiont is much higher than those of the endosymbionts in related aphid species reported so far. It appears either that the A. kondoi endosymbiont is derived from an ancestral bacterium different from those in other aphids or that its G+C content increased in a relatively short time after the evolutionary divergence of its host.

Amino Acid Sequence↗

Characteristic features of the nucleotide sequences of yeast mitochondrial ribosomal protein genes as analyzed by computer program GeneMark.

The nucleotide sequence data for yeast mitochondrial ribosomal protein (MRP) genes were analyzed by the computer program GeneMark which predicts the presence of likely genes in sequence data by calculating statistical biases in the appearance of consecutive nucleotides. The program uses a set of standard sequence data for this calculation. We used this program for the analysis of yeast nucleotide sequence data containing MRP genes, hoping to obtain information as to whether they share features in common that are different from other yeast genes. Sequence data sets for ordinary yeast genes and for 27 known MRP genes were used. The MRP genes were nicely predicted as likely genes regardless of the data sets used, whereas other yeast genes were predicted to be likely genes only when the data set for ordinary yeast genes was used. The assembled sequence data for chromosomes II, III, VIII and XI as well as the segmented data for chromosome V were analyzed in a similar manner. In addition to the known MRP genes, eleven ORF's were predicted to be likely MRP genes. Thus, the method seems very powerful in analyzing genes of heterologous origins.

Base Sequence↗

High frequency of cancer patients with abnormal assembly of the T cell receptor-CD3 complex in peripheral blood T lymphocytes.

Structural abnormality of T cell receptor (TCR)-CD3 complex on the cell surface was investigated in peripheral blood lymphocytes (PBL) from 55 cancer patients. In 24 of the 68 tests done on these patients, the CD3 zeta chain was not detected by immunoprecipitation with anti-CD3 epsilon monoclonal antibody (mAb), but was observed with anti-CD3 zeta mAb, suggesting that a high frequency of cancer patients possesses abnormal T cell receptor (TCR) complex in PBL. On the other hand, the total zeta chain was missing in several advanced cases. During follow-up of several patients, the zeta chain became undetectable after two or three months of cancer progression. It appears that immunosuppressive status can be monitored by analyzing the TCR-CD3 complex on the cell surface of PBL.

Disease Progression↗

Stimulation by menthol of Cl secretion via a Ca(2+)-dependent mechanism in canine airway epithelium.

1. To investigate the effect of menthol on airway epithelial ion transport function, we studied the bioelectrical properties of canine cultured tracheal epithelium by Ussing's short-circuit technique in vitro. 2. Addition of menthol (10(-3) M) to the mucosal but not the submucosal solution increased the short-circuit current (Isc) from 6.2 +/- 0.9 to 14.0 +/- 2.2 microA cm-2 (P < 0.001), and this effect was accompanied by increases in transepithelial potential difference and conductance. The response was dose-dependent, with the maximal increase from the baseline value and the concentration required to produce a half-maximal effect (EC50) being 6.4 +/- 0.9 microA cm-2 (P < 0.001) and 40 microM, respectively. 3. Other cyclic alcohols, including menthone and cyclohexanol, had no effect on the electrical properties. 4. The menthol-induced increase in Isc was not altered by pretreatment of the cells with amiloride, indomethacin, or propranolol but was abolished by diphenylamine-2-carboxylate, furosemide or substitution of Cl with iodide in the medium. 5. Menthol (10(-3) M) increased cytosolic levels of free calcium ([Ca2+]i) from 98 +/- 12 to 340 +/- 49 nM (P < 0.01) in fura-2-loaded tracheal epithelium but did not affect the intracellular adenosine 3',5'-cyclic monophosphate content. 6. These results suggest that menthol stimulates Cl secretion across airway epithelium, probably through a Ca(2+)-dependent mechanism, and might thus influence mucociliary transport in the respiratory tract.

Animals↗

Angiotensin II 1 receptor-mediated contraction of pulmonary artery and its modulation by prolylcarboxypeptidase.

To determine the subtype of angiotensin II (ANG II) receptor involved in the contraction of pulmonary artery and to elucidate its possible modulation by endogenous peptidases, we studied canine isolated pulmonary arterial rings under isometric conditions in vitro. Addition of ANG II caused a concentration-dependent contraction, an effect that was not altered by the ANG II 2 receptor antagonist EXP655 but was depressed by the ANG II 1 receptor antagonist DuP 753 so that the ANG II response curves were displaced to higher concentration by 1.5-2.0 log U (P < 0.001). Pretreatment of tissues with the prolylcarboxypeptidase (PCP) inhibitor p-methylphenyl sulfonyl-fluoride potentiated the ANG II-induced contraction, with the concentration required to produce a half-maximal effect of ANG II being decreased from 4.1 +/- 0.9 x 10(-9) to 3.8 +/- 0.5 x 10(-10) M (P < 0.001), whereas other peptidase inhibitors such as p-chloromercuriphenyl sulfonic acid, amastatin, and phosphoramidon had no effect. The p-methylphenyl sulfonylfluoride-induced potentiation was abolished by the removal of endothelium, but it was still observed in the presence of NG-nitro-L-arginine methyl ester in the endothelium-intact tissues. The PCP activity in the tissues was reduced by the removal of endothelium from 645 +/- 88 to 91 +/- 29 nmol.mg protein-1.h-1 (P < 0.001), and cultured endothelium had the activity of 404 +/- 39 nmol.mg protein-1.h-1. These results suggest that ANG II contracts pulmonary artery via ANG II 1 receptor and that PCP localized to the endothelium may have a modulatory role in the ANG II-induced pulmonary vasoconstriction.

Angiotensin II↗

Effect of platelet-activating factor on intracellular free calcium in cow tracheal epithelium.

The effect of platelet activating factor (PAF) on the intracellular cytosolic levels of free calcium ([Ca2+]i) was studied in cultured epithelium from cow trachea. In fura-2-loaded cells, PAF (10(-9) to 10(-5) M), but not lyso-PAF, increased [Ca2+]i in a concentration-dependent manner, from 106 +/- 15 to 270 +/- 40 nM (P < 0.05). This [Ca2+]i response consisted of a transient increase that peaked within 15 s after addition and a subsequent sustained elevation that reached a plateau after 1 min. The potency for the sustained response was greater by approximately 1 log U than that for the transient response. Preincubation of the cells with the PAF receptor antagonist CV6209 (10(-6) M) inhibited the increase in [Ca2+]i. Ca(2+)-free medium (2 mM EGTA) totally abolished the sustained response to PAF, but it only partially inhibited the transient response. Verapamil (10(-5) M) also largely inhibited the sustained response. Moreover, PAF transiently increased inositol triphosphate (IP3) levels, peaking 10 s after addition. These data suggest that (1) a PAF-induced increase in [Ca2+]i is mediated via PAF receptors, (2) PAF causes both transient [Ca2+]i release from intracellular stores through IP3 production and sustained [Ca2+]i influx from extracellular solution, and (3) Ca2+ influx may be a major pathway of PAF-induced increase in [Ca2+]i in cow tracheal epithelium.

Animals↗

Inhibitory action of epiderstatin on EGF-stimulated growth of mouse epidermal. BALB/MK cells without direct effect on protein kinase activities.

Epiderstatin, a distinctive glutarimide antibiotic isolated from Streptomyces pulveraceus subsp. epiderstagenes, has been revealed to be a potent inhibitor of the signal transduction of epidermal growth factor (EGF). Epiderstatin inhibited the DNA synthesis induced by various peptide growth factors in a mouse epidermal cell line, BALB/MK, without inhibiting protein tyrosine kinase activity of EGF-receptor or serine/threonine kinase activity of protein kinase C. The 50% inhibitory concentration (IC50) value of epiderstatin for the EGF-stimulated incorporation of [3H]thymidine into BALB/MK cells was about 10 nM. When epiderstatin was added to the quiescent cells simultaneously with EGF-stimulation, the cells did not reenter into the growing cell cycle. The action of epiderstatin proceeded from the overexpression of c-fos and the suppression of c-myc transcription when EGF was added to quiescent BALB/MK cells.

Actins↗

Signaling pathway other than phosphatidylinositol turnover is responsible for constant expression of c-myc gene in primary cultures of rat hepatocytes.

We investigated signal transduction pathways involved in c-myc activation, using rat hepatocytes in primary culture. c-Myc mRNA was constantly expressed in the cultured hepatocytes regardless of the conditions present. When the expression was examined in the presence of various agents modulating intracellular signals, isoflavonoids (genistein, psi-tectorigenin, and orobol) significantly decreased c-myc mRNA levels, in a dose dependent manner. However, genistein did not decrease Li+ induced inositol phosphate accumulation using [3H]inositol-labeled cultured hepatocytes. In addition, we have shown that these isoflavonoids increase cytoplasmic free Ca2+, when measured using aequorin-loaded hepatocytes. In light of these observations, the persistent basal level of c-myc expression seems to be maintained by mechanisms other than phosphatidylinositol turnover.

Animals↗

Alcohol consumption and risk of esophageal cancer in Japan: a case-control study in seven hospitals.

In a multi-center case-control study, we evaluated the risk of esophageal cancer in the Japanese population. All patients and controls were inpatients in the surgical departments of seven hospitals nationwide. Patients eligible for the study were those newly diagnosed as having primary esophageal cancer. One control per case was selected from among patients admitted to the same hospital, and 141 male pairs were analyzed using logistic regression analysis. The results showed dose-response relation between the risk of esophageal cancer and both the quantity (g/week) and frequency (times/week) of alcohol drinking (P value for trend = 0.0001). Although a statistically significant risk increase was shown among moderate to heavy smokers (15 < or = cigarette/day < 25) (odds ratio, 4.35:95% confidence interval, 1.81-10.49), the dose-response for cigarette smoking was unclear (P value for trend = 0.07). No combined effect of alcohol drinking and cigarette smoking was found. A frequent intake of fruit was associated with a decreased risk (P value for trend = 0.02). After adjustment for alcohol consumption, cigarette smoking and fruit intake were found not to be associated with the risk, whereas a preference for high-temperature food and drink showed a statistically significant positive association (P value for trend = 0.02). Drinkers who consumed shochu most frequently showed a three-fold increased risk over that for beer consumers, although the association disappeared after adjusting for the amount of alcohol consumed. The present results confirm alcohol intake and a preference for high-temperature food to be associated with an increased risk of esophageal cancer and show the amount of alcohol consumed, rather than the type of alcoholic beverage, to be the main risk determinant.

Alcohol Drinking↗

[Analysis of glucose metabolism in patients with esophageal cancer by PET: estimation of hexokinase activity in the tumor and usefulness for clinical assessment using 18F-fluorodeoxyglucose].

We evaluated glucose metabolism of esophageal cancer by PET using 18F-fluorodeoxyglucose (FDG), in order to investigate its clinical usefulness. In 11 advanced cases k3 value reflecting hexokinase activity and Ci/Cp ratio expressing FDG uptake were calculated from radioactivity in the tumor (Ci) and the plasma (Cp). Both k3 and Ci/Cp were well correlated with hexokinase activity from the resected specimen, so Ci/Cp was considered to be a convenient index for clinical assessment of esophageal cancer. Forty-two cases before treatment revealed high accumulation of FDG, and 41 showed more than 2.0 of Ci/Cp. But, 10 normal controls and one esophageal benign tumor showed less than 2.0. As for 13 post-operative cases, 6 cases out of 7 with recurrence showed more than 2.0 for Ci/Cp, but all 6 cases of non-recurrent cases showed less than 2.0. FDG PET is an useful tool for differential diagnosis of recurrence. The clinicopathological findings were investigated in 26 resected cases. Age, location, vertical extension, histologic feature, lymph node metastasis and histologic stage were not correlative with Ci/Cp. As for DNA ploidy pattern, aneuploidy group showed significantly high Ci/Cp rather than diploidy group. Eight cases which showed more than 5.0 of Ci/Cp resulted in poor prognoses compared with 14 cases which showed less than 5.0.

Aged↗

[Modulation of cholinergic neurotransmission by Ca(2+)-activated K channel and Na(+)-K(+)-ATPase in canine airway smooth muscle].

Stimulation of Ca(2+)-activated K+ channel and Na(+)-K(+)-ATPase may play an important role in the relaxant responses of airway smooth muscle to certain bronchodilators. To elucidate whether cholinergic neuroeffector transmission can be modulated by Ca(2+)-activated K+ channel and Na(+)-K(+)-ATPase, we studied canine airway smooth muscle under isometric conditions in vitro. Addition of charybdotoxin (ChTx, 10(-7) M) did not alter the contractile responses to acetylcholine (ACh) but augmented electrical field stimulation (FES)-induced contractions at 1-10 Hz (p < 0.01), whereas apamin and glibenclamide were without effect. This effect was dose-dependent, with the maximal increase being 36.8 +/- 5.3% (p < 0.001). Ouabain at a concentration insufficient to alter the resting tension (10(-7) M) increased contractions induced by both EFS and ACh. The magnitude of the increase in contractile responses to EFS was similar to those to ACh at ouabain concentration of up to 3 x 10(-7) M, but the former was significantly greater at 10(-6) M ouabain (p < 0.05). These results suggest that each Ca(2+)-activated K+ channel and Na(+)-K(+)-ATPase may a modulatory role in the cholinergic neurotransmission by inhibiting the exocytotic ACh release from the vagal nerve terminals.

Acetylcholine↗

[Evaluation of clinical usefulness of 11C-methionine positron emission tomography (11C-MET-PET) as a tool for liver functional imaging].

We studied 11C-MET-PET in 17 clinical cases, 10 patients with obstructive jaundice and 7 normal volunteers, and analyzed its efficacy for the evaluation of hepatic functional reserve in major hepatectomy candidates. Differential absorption ratio (DAR) of 11C was compared to the hepatic protein synthesis rate (HPS), which is measured as the incorporation rate of 3H-labeled leucine in protein fraction, using needle biopsied liver specimen obtained from each hepatic segment. In the cases of normal liver function, DAR was well correlated with HPS. Also in jaundice cases with two exceptions, low HPS segment was demonstrated as low DAR segment. Consequently, MET-PET images could clearly provide functional liver imaging. After injection to 11C-MET, the increase in rate of radioactivity of 11C in plasma protein fraction was higher in jaundice cases than in normal volunteers, which is in accord with the results of our former study that cholestatic liver has accelerated protein synthesis rate. In summary, since 11C-MET-PET could demonstrate liver functional imaging, it might be a possible tool for liver function assessment in major hepatectomy candidates.

Carbon Radioisotopes↗