Biomedical subjects

Y Okamura

Publications and source records attributed to Y Okamura.

At least 37 records · Page 2Linked to original sources

PubMed2001

Changes in gelsolin expression during ascidian metamorphosis.

Gelsolin is an actin regulatory protein that is expressed in a wide variety of tissues and is especially abundant in muscle and blood cells. The role of gelsolin during structural reorganization of the body, such as during metamorphosis or regeneration, is poorly understood. We analyzed changes in gelsolin expression during ascidian embryogenesis and metamorphosis using nucleic acid probes and a monoclonal antibody (AS23) specific for ascidian gelsolin; our results indicated that gelsolin is maternally provided and that its de novo gene transcription is initiated during the neurula stage. In the larva, gelsolin was detectable in specific types of nerve cells, i.e. the adhesive papillae, motor neurons and epidermal sensory neurons. During metamorphosis, the expression of gelsolin changes markedly: the expression is suppressed in nerve tissues after tail resorption but is induced in mesodermal tissues. Gelsolin accumulated in mesenchyme cells until the onset of tail resorption, and following tail resorption, these cells migrated to the tunic and differentiated into tunic cells with many fine processes. Migration of the mesenchyme cells into the tunic was completely inhibited by treatment with cytochalasin B. Gelsolin was colocalized with actin in tunic cells, suggesting that it is involved in the rearrangement of actin filaments during cell locomotion or morphogenesis.

Animals↗

PubMed2001

The larval ascidian nervous system: the chordate brain from its small beginnings.

The body plan of the tadpole larva of ascidians, or sea-squirts, is widely presumed to be close to that of the hypothetical ancestor of all chordate animal groups, including vertebrates. This is nowhere more obvious than in the organization and development of the dorsal tubular nervous system. Several recent developments advocate this model neural system for studies on neurobiology and neurogenesis. These include advances in our understanding of development in ascidian embryos and of differentiation among the cellular progeny of its neural plate; the application of transgenic and mutant approaches to studies on ascidian larval neurones; and the prospect of advances in genomic analyses. In addition to providing ways to study a working chordate brain in miniature, all these offer insights into the ancestral condition of the developing vertebrate brain.

Animals↗

PubMed2001

Mössbauer study of the reaction of laser-evaporated iron atoms with nitrogen molecules in low-temperature argon matrices.

Reaction products of laser-evaporated iron atoms with nitrogen molecules were investigated using a low-temperature matrix isolation technique. The products observed in the Mössbauer spectra were FeNNN, Fe(NN), and Fe(NN)2, isolated in argon matrices. Yields of the species changed, depending on the concentration of nitrogen in the matrix. A product assigned to Fe(NN)6 was obtained in a pure nitrogen matrix. Infrared spectra and molecular orbital calculations of the products support these findings.

Journal Article↗

PubMed2001

Fibrinogen-conjugated albumin polymers and their interaction with platelets under flow conditions.

Albumin polymers, having an average diameter of 1020 +/- 250 nm, were prepared by the disulfide polymerization of recombinant human serum albumin (rHSA) by controlling of the pH and temperature. Fibrinogen could be conjugated on the surface of an albumin polymer using N-succinimidyl 3-(2-pyridyldithio)propionate (SPDP). Under flow conditions, the fibrinogen-conjugated albumin polymers (fibrinogen-albumin polymers) were irreversibly attached to the platelet-immobilized surface in the reconstituted blood at a low platelet concentration ([platelet] = 5.0 x 10(4)/microL, a 5-fold diluted platelet concentration), and the attachment was suppressed by the addition of anti-GPIIb/IIIa monoclonal antibodies. It was confirmed that fibrinogen-albumin polymers specifically interacted with GPIIb/IIIa expressed on the surface of the activated platelets. Although platelets with a low platelet concentration were hardly attached to the platelet-immobilized surface under the flow conditions, the addition of fibrinogen-albumin polymers enhanced the attachment of the remaining platelets to the surface, indicating that the fibrinogen-albumin polymers would help the hemostatic ability of platelets at the site of vascular injury of patients in thrombocytopenia.

Biocompatible Materials↗

PubMed2001

Effects of idoxifene and estradiol on NF-kappaB activation in cultured rat hepatocytes undergoing oxidative stress.

BACKGROUND/AIMS: Idoxifene is a tissue-specific selective estrogen receptor modulator. Estradiol is a potent endogenous antioxidant, and nuclear factor kappaB (NF-kappaB) is a key transcription factor that induces multiple genes in response to inflammation or oxidative stress. The aim of this study was to explore the inhibitory effects of idoxifene and estradiol on NF-kappaB activation in hepatocytes in a state of oxidative stress. METHODS: Lipid peroxidation was induced in cultured rat hepatocytes by incubation with ferric nitrilotriacetate solution. NF-kappaB activity was evaluated by electrophoretic mobility shift assay. RESULTS: The oxidative stress-induced activation of NF-kappaB and degradation of IkappaB-alpha were maximal at 3-5 h, with an increase in lactate dehydrogenase (LDH) and malondialdehyde (MDA) secretion into the culture medium. Treatment with idoxifene and estradiol inhibited IkappaB-alpha degradation and NF-kappaB activation through the attenuation of hepatocyte oxidative bursts and decreased extracellular levels of LDH and MDA. In addition, idoxifene and estradiol inhibited lipid peroxidation in rat liver mitochondria. A potent NF-kappaB inhibitor, pyrrolidine dithiocarbamate, prevented NF-kappaB activation by inhibition of IkappaB-alpha degradation and decreased LDH and MDA levels, suggesting that NF-kappaB might be a regulator in a genetic response to increase oxidative stress-induced hepatic injury. CONCLUSIONS: These findings suggest that idoxifene and estradiol function as antioxidants and protect hepatocytes from inflammatory cell injury.

Animals↗

PubMed2001

Protein tyrosine kinase expression in the porcine ovary.

Various growth factor receptors contain intrinsic tyrosine kinase activity, indicating that protein tyrosine kinases (PTK) play an important role in signal transduction pathways for cell proliferation and differentiation. To identify oocyte-derived factors which control follicle cells as well as oocyte-controlling factors produced by follicle cells, we examined the expression of genes which contain the PTK domain in the porcine ovary, using a polymerase chain reaction-based amplification technique with degenerate oligonucleotide primers that are specific to the PTK domain. Clones for the porcine homologues of platelet-derived growth factor receptor alpha (PDGFRalpha) and of insulin-like growth factor-I receptor (IGF-IR) were found during follicle growth both in oocytes and follicle cells. Clones for the porcine homologues of focal adhesion kinase (FAK), of c-kit and of fms-like tyrosine kinase (FLT)-3 were found only in oocytes. Moreover, after 24 h of in-vitro maturation of the cumulus-oocyte complexes, clones for the porcine homologues of FLT-1, of FLT-4, of Tie2 and of RYK in oocytes were observed. Immunohistochemical studies revealed the existence of PDGFRalpha, platelet-derived growth factor A (PDGFA), FAK and FLT3 in oocytes at various stages of folliculogenesis. These results suggest that fluctuations in the expression of these PTK genes may be involved in follicle growth and maturation.

Amino Acid Sequence↗

PubMed2001

How is the liver primed or sensitized for alcoholic liver disease?

This article represents the proceedings of a symposium at the 2000 ISBRA Meeting in Yokohama, Japan. The chairs were Hidekazu Tsukamoto and Yoshiyuki Takei. The presentations were (1) Tribute to Professor Rajendar K. Chawla, by Craig J. McClain; (2) Dysregulated TNF signaling in alcoholic liver disease, by Craig J. McClain, S. Joshi-Barve, D. Hill, J Schmidt, I. Deaciuc, and S. Barve; (3) The role of mitochondria in ethanol-mediated sensitization of the liver, by Anna Colell, Carmen Garcia-Ruiz, Neil Kaplowitz, and Jose C. Fernandez-Checa; (4) A peroxisome proliferator (bezafibrate) can prevent superoxide anion release into hepatic sinusoid after acute ethanol administration, by Hirokazu Yokoyama, Yukishige Okamura, Yuji Nakamura, and Hiromasa Ishii; (5) S-adenosylmethionine affects tumor necrosis factor-alpha gene expression in macrophages, by Rajendar K. Chawla, S. Barve, S. Joshi-Barve, W. Watson, W. Nelson, and C. McClain; (6) Iron, retinoic acid and hepatic macrophage TNFalpha gene expression in ALD, by Hidekazu Tsukamoto, Min Lin, Mitsuru Ohata, and Kenta Motomura; and (7) Role of Kupffer cells and gut-derived endotoxin in alcoholic liver injury, by N. Enomoto, K. Ikejima, T. Kitamura, H. Oide, Y. Takei, M. Hirose, B. U. Bradford, C. A. Rivera, H. Kono, S. Peter, S. Yamashina, A. Konno, M. Ishikawa, H. Shimizu, N. Sato, and R. Thurman.

Animals↗

PubMed2001

Neuronal form in the central nervous system of the tadpole larva of the ascidian Ciona intestinalis.

The dorsal tubular central nervous system (CNS) of the ascidian tadpole larva is a diagnostic feature by which the chordate affinities of this group, as a whole, are recognized. We have used two methods to identify larval neurons of Ciona intestinalis. The first is serial electron microscopy (EM), as part of a dedicated study of the visceral ganglion (1), and the second is the transient transfection of neural plate progeny with green fluorescent protein (GFP) (2), to visualize the soma and its neurites of individual neurons in whole-mounted larvae of C. intestinalis. Our observations reveal that ascidian larval neurons are simple inform, with a single axonal neurite arising from a soma that is either monopolar or has only very few, relatively simple neurites arising from it, as part of a presumed dendritic arbor. Somata in the visceral ganglion giving rise to axons descending in the caudal nerve cord are presumed to be those of motor neurons.

Animals↗

PubMed2001

[Usefulness of OPCAB from the viewpoint of fluctuations in the level of blood natriuretic peptides].

Fluctuations in the level of blood natriuretic peptides (ANP and BNP) were compared between 41 patients who underwent conventional coronary artery bypass (CCAB) and 19 patients who underwent off-pump coronary artery bypass (OPCAB). A blood sample was collected before surgery, and 6, 12 and 24 hours; 2, 3, 5 and 7 days; and 1 month after the end of extracorporeal circulation. There were no significant differences in left ventricular ejection fraction (LVEF) before and after surgery in either group or between the two groups. On average, 3.3 +/- 1.0 bypass grafts were used for the CCAB group, and 2.2 +/- 0.8 grafts for the OPCAB group. Furthermore, the maximum postoperative creatine phosphokinase-MB (CK-MB) level for the CCAB group was 49.1 +/- 17.5 IU/l, whereas that for the OPCAB group was significantly lower at 23.2 +/- 24.8 IU/l. The preoperative level of blood ANP for the CCAB group was 24.6 +/- 19.9 pg/ml while that for the OPCAB group was 39.3 +/- 29.5 pg/ml, but there was no significant difference between the two groups. In both groups, the level of blood ANP reached a peak three days after the end of extracorporeal circulation and then decreased after that point. Although the level of blood ANP for the OPCAB group decreased to 51.4 +/- 26.4 pg/ml one month after the end of extracorporeal circulation, that for the CCAB group one month after the end of extracorporeal circulation remained significantly high at 61.3 +/- 30.6 pg/ml, when compared to that before surgery. Furthermore, the preoperative level of blood BNP for the CCAB group was 40.0 +/- 35.2 pg/ml and that for the OPCAB group was 75.5 +/- 59.7 pg/ml, but there was no significant difference between the two groups. Then, in both groups, the level of blood BNP reached a peak 2-5 days after the end of extracorporeal circulation and then decreased after that. Whereas the level of blood BNP for the OPCAB group decreased to 96.4 +/- 56.0 pg/ml one month after the end of extracorporeal circulation, that for the CCAB group one month after the end of extracorporeal circulation remained significantly high at 160.3 +/- 106.2 pg/ml when compared to that before surgery. The levels of ANP and BNP increased postoperatively for both OPCAB and CCAB groups since the following events caused a great degree of stress on the heart: general anesthesia, cardiac herniation, stabilizer compression, regional blood flow blockage and reperfusion injury. Although the level of natriuretic peptides for the CCAB group remained high one month after the end of surgical stress, that for the OPCAB group returned to near the preoperative level one month later, thus supporting the notion that OPCAB is less invasive.

Aged↗

PubMed2001

Coronary artery bypass in dialysis patients.

This study evaluated the operative outcome of dialysis patients undergoing coronary artery bypass (CAB). In the past 6 years, 38 dialysis patients with a mean age of 57.5 years underwent CAB. Thirty-one operations were elective, and 7 were nonelective operations. Thirty-two operations were performed under cardiac arrest, 3 operations were performed under fibrillatory arrest, and 3 operations were performed without cardiopulmonary bypass. The average number of bypass grafts was 2.8. In all patients, the internal thoracic artery was used. Hospital mortality was 5.3%. Actuarial survival rates at 1, 3, and 5 years including all causes of death were 88%, 80%, and 72%, respectively. With improvements in perioperative management, coronary artery bypass can be performed with acceptable mortality and morbidity in dialysis patients. Complete revascularization without cardiopulmonary bypass is the ideal method and will increase in usage. However, the conventional CAB also provides acceptable results.

Adult↗

PubMed2001

Primary structure, developmental expression and functional properties of an inward rectifier K+ channel of the tunicate.

A cDNA encoding for a tunicate inward rectifier K+ channel (TuIRK) was isolated. TuIRK exhibited the highest similarity (approximately 50%) with mammalian Kir2 (IRK) subfamily. Maternal RNA of TuIRK was detected by RT-PCR in unfertilized eggs. By in situ hybridization, the transcript was observed at the 32-cell stage, restricted at the 64-cell stage in anterior epidermal cells of a4-2 blastomere lineage, and disappeared at the late gastrula stage. Therefore, TuIRK was identified to be the inward rectifier whose expression was previously reported to change dramatically upon the neural/epidermal cell fate selection. In Xenopus oocytes, TuIRK expressed a strongly inward rectifying K+ current. The basic electrophysiological properties of TuIRK were similar to those of the mouse IRK1 (mIRK1), except that the sensitivity to the block by extracellular Mg2+ was much lower than that of mIRK1. To identify the structural determinant, we made mutants of the pore region, and then of the extracellular loop (N226 of TuIRK, and E125 of mIRK1). In E125N mutant of mIRK1, the sensitivity to the Mg2+ block was decreased significantly, whereas N226E of TuIRK1 did not acquire the sensitivity. These results demonstrate the contribution of this site to the Mg2+ block and the presence of additional determinant(s).

Amino Acid Sequence↗

PubMed2000

Serine residue in the IIIS5-S6 linker of the L-type Ca2+ channel alpha 1C subunit is the critical determinant of the action of dihydropyridine Ca2+ channel agonists.

The dihydropyridine (DHP)-binding site has been identified within L-type Ca(2+) channel alpha(1C) subunit. However, the molecular mechanism underlying modulation of Ca(2+) channel gating by DHPs has not been clarified. To search for novel determinants of high affinity DHP binding, we introduced point mutations in the rat brain Ca(2+) channel alpha(1C) subunit (rbCII or Ca(v)1.2c) based on the comparison of amino acid sequences between rbCII and the ascidian L-type Ca(2+) channel alpha(1) subunit, which is insensitive to DHPs. The alpha(1C) mutants (S1115A, S1146A, and A1420S) and rbCII were transiently expressed in BHK6 cells with beta(1a) and alpha(2)/delta subunits. The mutation did not affect the electrophysiological properties of the Ca(2+) channel, or the voltage- and concentration-dependent block of Ca(2+) channel currents produced by diltiazem and verapamil. However, the S1115A channel was significantly less sensitive to DHP antagonists. Interestingly, in the S1115A channel, DHP agonists failed to enhance whole-cell Ca(2+) channel currents and the prolongation of mean open time, as well as the increment of NP(o). Responsiveness to the non-DHP agonist FPL-64176 was also markedly reduced in the S1115A channel. When S1115 was replaced by other amino acids (S1115D, S1115T, or S1115V), only S1115T was slightly sensitive to S-(-)-Bay K 8644. These results indicate that the hydroxyl group of Ser(1115) in IIIS5-S6 linker of the L-type Ca(2+) channel alpha(1C) subunit plays a critical role in DHP binding and in the action of DHP Ca(2+) channel agonists.

Amino Acid Sequence↗

PubMed2000

Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization.

A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method.

DNA Probes↗

PubMed2000

Diversity of voltage-gated sodium channels in the ascidian larval nervous system.

To gain insight into the origin of the molecular diversity of voltage-gated sodium channels (NaVs), a putative sodium channel gene (TuNa2) was cloned from the protochordate ascidian. TuNa2 showed two unusual features in its primary structure; (1) lysine in the P-region of the third repeat, a critical site determining ion selectivity, was changed to glutamic acid, predicting that the ionic permeability would not be rigidly sodium-selective (2) the III-IV linker, determinant of fast inactivation, was only weakly conserved. In contrast with a pan-neuronally expressed NaV (TuNa1), expression of TuNa2 was confined to subsets of neurons including motor neurons, suggesting that TuNa2 plays specialized roles in electrical activities unique to these neurons. Basic FGF, a neural inducer in the ascidian embryo, induces TuNa2 RNA expression in the ectodermal cells at lower doses than that required for TuNa1 gene expression. Thus, two types of NaV may play distinct roles and their gene expressions are controlled by distinct mechanisms.

Amino Acid Sequence↗

PubMed2000

Cloning and characterization of a gene, mpsA, encoding a protein associated with intracellular magnetic particles from Magnetospirillum sp. strain AMB-1.

Proteins located within the lipid bilayer, surrounding the intracellular bacterial magnetic particles (BMP) from Magnetospirillum sp. AMB-1, were separated using SDS-PAGE. Several major proteins of approximate molecular weight 66.2, 35.6, and 24.8 kDa were identified. The N-terminal amino acid sequence of one of these proteins, designated MpsA, was determined and used to design a pair of PCR primers which amplified a 105 bp DNA fragment from AMB-1 genomic DNA. Gene-walking, using anchored PCR, was used to determine the complete nucleotide sequence (954 bp) of the mpsA gene. The mpsA encodes a 317 amino acid protein which does not have an N-terminal cytoplasmic transport signal sequence. Intracellular localization studies were carried out using an mpsA-luc gene fusion expressed in AMB-1 following gene transfer by conjugation. The gene fusion was constructed by cloning a 1.6 kb mpsA fragment upstream of luc in the conjugal plasmid pKLC. The MpsA-Luc fusion protein was preferentially located on the magnetic particle membrane. Although the function of MpsA remains unknown, homology searches suggest similarity with the alpha subunit of acetyl-CoA carboxylase and the CoA-binding motif.

Amino Acid Sequence↗

PubMed2000

Histopathological and physiological characteristics of cultured human ACTH-secreting cells derived from a rapidly growing pituitary adenoma.

We observed the histopathological and physiological characteristics of adrenocorticotropic hormone (ACTH)-secreting adenoma cells derived from a rapidly growing pituitary adenoma, which have firm cell attachment and well-preserved hormonal function in a relatively longterm culture. Corticotrophs, obtained from a 43-year-old woman with Cushing's disease in whom plasma ACTH levels increased in response to 1-deamino-8-D-arginine vasopressin (DDAVP) stimulation and the proliferative potential was very high, were grown in tissue culture for up to 6 months. The morphological features were observed by phase contrast and electron microscopy. The cultured cells were incubated with corticotroph-releasing hormone (CRH), arginine vasopressin (AVP), or DDAVP, and ACTH in the medium was measured by radioimmunoassay (RIA). The morphology of the ACTH-secreting adenoma cells in culture revealed a mixed population of formed clusters and spindle-shaped fibroblast-like cells. The adenoma cells were immunohistochemically positive only for ACTH. On electron microscopic observation, pituitary tumor cells obtained 6 days after seeding demonstrated many secretory granules, well-developed rough endoplasmic reticulum, and mitochondria; fewer secretory granules were observed after cultivation for 24 days. ACTH levels in the incubation media were elevated with stimulation by DDAVP, AVP, or CRH. In this study, the establishment of relatively longterm culture of human pituitary adenoma cells seemed to be due to the high proliferative potential of this adenoma. This in vitro study may imply that DDAVP as well as AVP directly stimulates ACTH release from corticotropic adenoma cells.

Adenoma↗

PubMed2000

Mössbauer studies on laser evaporated iron atoms and their reactions with oxygen in argon matrices.

Laser-evaporated iron atoms were isolated in low-temperature Ar matrices and their chemical reactions with oxygen were investigated by means of Mössbauer spectroscopy. Reactions of iron atoms with oxygen produce FeO, Fe(O2), FeO3, (O2)FeO2 and OFeO isolated in the Ar matrices and their yields vary depending on the concentration of oxygen. Similarly, FeO and Fe(O2) were obtained by the reaction of iron atoms with N2O. Infrared spectroscopy and molecular orbital calculations were applied to support their assignments.

Calorimetry↗

PubMed2000

Pharmacokinetics of the anticoagulant 14C-DX-9065a in the healthy male volunteer after a single intravenous dose.

1. The plasma pharmacokinetics, excretion and metabolism of DX-9065a were studied in the healthy male Caucasian volunteer after a single intravenous dose of 10 mg 14C-labelled DX-9065a. 2. At the end of a 1 h infusion, the mean plasma concentration of total radioactivity was 380 ng ml(-1) (equivalent to unchanged DX-9065). Thereafter, it decreased in a bi-exponential manner and was below the detection limit by 48 h after dosing. The half-life for the distribution phase was 6.93 h. 3. The total radioactivity recovered in urine and faeces by 336 h post-dose was 83.8% of the administered dose, with excretion ongoing at the end of the 14-day collection. The major route of excretion was via urine, accounting for a mean of 77.6% of the administered radioactivity. The urinary excretion profile was biphasic, consisting of rapid (0-24 h) and slow (24-336 h) phases. A large renal clearance suggested that renal tubular secretion might contribute to the excretion of DX-9065 via urine. 4. No metabolite peaks in the radio-HPLC chromatograms of urine samples were detected, indicating that biotransformation of DX-9065 does not play a significant role in the elimination of DX-9065 in man.

Adult↗