Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 217 records · Page 12Linked to original sources

PubMed1996

The basic amino acid transporter (rBAT)-like immunoreactivity in paraventricular and supraoptic magnocellular neurons of the rat hypothalamus.

In the rat hypothalamus, the basic amino acid transporter (rBAT)-like immunoreactivity was analyzed by immunohistochemistry using an antibody against the 15-amino acid sequence of the deduced rat rBAT protein. In the supraoptic and the paraventricular nuclei, magnocellular neurons exhibited the marked rBAT-like immunoreactivity in intracellular structures but not in the plasma membrane. The results suggest that the rBAT serves as an intracellular amino acid transport system in magnocellular neurons.

Amino Acid Sequence↗

PubMed1996

EGF or PDGF receptors activate atypical PKClambda through phosphatidylinositol 3-kinase.

Overexpression of a TPA-insensitive PKC member, an atypical protein kinase C (aPKClambda), results in an enhancement of the transcriptional activation of TPA response element (TRE) in cells stimulated with epidermal growth factor (EGF) or platelet-derived growth factor (PDGF). EGF or PDGF also caused a transient increase in the in vivo phosphorylation level and a change in the intracellular localization of aPKClambda from the nucleus to the cytosol, indicating the activation of aPKClambda in response to this growth factor stimulation. These immediate signal-dependent changes in aKPClambda were observed for a PDGF receptor add-back mutant (Y40/51) that possesses only two of the five major autophosphorylation sites and binds PI3-kinase, and were inhibited by wortmannin, an inhibitor of PI3-kinase. Furthermore, an N-terminal fragment of the catalytic subunit of PI3-kinase, p110alpha, inhibited aPKClambda-dependent activation of TRE in Y40/51 cells stimulated with PDGF. Overexpression of p110alpha resulted in an enhancement of TRE expression in response to PDGF and the regulatory domain of aPKClambda inhibited this TRE activation in Y40/51 cells. These results provide the first in vivo evidence supporting the presence of a novel signalling pathway from receptor tyrosine kinases to aPKClambda through PI3-kinase.

Amino Acid Sequence↗

PubMed1996

Platelet-derived growth factor activates protein kinase C epsilon through redundant and independent signaling pathways involving phospholipase C gamma or phosphatidylinositol 3-kinase.

Protein kinase C (PKC), a major cellular receptor for tumor-promoting phorbol esters and diacylglycerols (DGs), appears to be involved in a variety of cellular functions, although its activation mechanism in vivo is not yet fully understood. To evaluate the signaling pathways involved in the activation of PKC epsilon upon stimulation by platelet-derived growth factor (PDGF) receptor (PDGFR), we used a series of PDGFR "add-back" mutants. Activation of a PDGFR mutant (Y40/51) that binds and activates phosphatidylinositol 3-kinase (PI 3-kinase) caused translocation of PKC epsilon from the cytosol to the membrane in response to PDGF. A PDGFR mutant (Y1021) that binds and activates phospholipase C gamma (PLC gamma), but not PI 3-kinase, also caused the PDGF-dependent translocation of PKC epsilon. The translocation of PKC epsilon upon stimulation of PDGFR (Y40/51) was inhibited by wortmannin, an inhibitor of PI 3-kinase. Activation of PKC epsilon was further confirmed in terms of PKC epsilon-dependent expression of a phorbol 12-tetradecanoate 13-acetate response element (TRE)-luciferase reporter. Further, purified PKC epsilon was activated in vitro by either DG or synthetic phosphatidylinositol 3,4,5-trisphosphate. These results clearly demonstrate that PKC epsilon is activated through redundant and independent signaling pathways which most likely involve PLC gamma or PI 3-kinase in vivo and that PKC epsilon is one of the downstream mediators of PI 3-kinase whose downstream targets remain to be identified.

Cell Compartmentation↗

PubMed1996

In vivo dynamics of myosin II in Dictyostelium by fluorescent analogue cytochemistry.

We used fluorescent analogue cytochemistry to study in vivo dynamics of myosin II in Dictyostelium discoideum. We labeled myosin with biotin or tetramethyl-rhodamine iodoacetamide (IATR). The labeled myosin shows normal activities as reversible filament assembly and Ca2+ and actin-activatable Mg(2+)-ATPase. We used the biotin-myosin as a probe examining the effects of microinjection on the amoebae and the ability to associate with endogenous actin cytoskeleton. The biotin-myosin incorporates into certain actin populations and localizes to the cortex with the highest accumulation in the posterior end of polarized amoebae. The dynamics in live amoebae were probed by TR-myosin. We monitored the dynamics for a long period to determined the dynamic reorganization corresponding specific cellular behaviors. The TR-myosin converges into a discrete actin- and myosin-rich structure located at the posterior end ("myosin-organizing center"). The rod-shaped TR-myosin exhibits linear orderly arrays emanating from the organizing center which extend about two-thirds of the cell length. The myosin arrays show a dynamic reorganization when the amoebae move. To examine if the observed myosin dynamics are related to filamentous (F-) actin, we disrupted the F-actin by cytochalasin D. The ratioed image of TR-myosin (vs. FITC-dextran) demonstrates that myosin in these cells accumulates in the cortex but does not form the organizing center. Overall, the results suggest that the filamentous myosin organizes into orderly arrays in the live cytoplasm and its translocation occurs by means of F-actin cables, converging into the organizing center.

Animals↗

PubMed1996

Morphological study of the human maxillofacial venous vasculature: examination of venous valves using the corrosion resin cast technique.

BACKGROUND: The venous drainage in the human head and neck, structures located superior to the heart, generally does not counter the force of gravity. Accordingly, venous valves in the human head have been reported to be few or nonexistent, and therefore the direction of venous drainage is variable and obscure. METHODS: We investigated whether venous valves were present in 13 heads of human adult cadavers using scanning electron microscopic observation of corrosive resin casts in the human maxillofacial region. This technique afforded the observation of the venous valves. RESULTS: Venous valves were found throughout the facial region. In particular, there were many valves distributed in the lingual, upper and lower labial, facial and pharyngeal veins. The venous valves were particularly well formed at motile parts of the maxillofacial region. CONCLUSIONS: It is suggested that these valves may aid the venous blood return and permit the blood drainage in one direction.

Aged↗

PubMed1996

Effects of cocaine on the rat cerebral commissure.

We investigated the effects of cocaine on the corpus callosum, the nerve fibre bundle that connects the bilateral cerebral hemispheres. Our experiments in rats confirmed that, in the control group, the mid-sagittal area of the corpus callosum in the adult male was significantly larger than this area in the female. Early postnatal exposure to cocaine abolished this sexual dimorphism, that is, cocaine-treated males had a significantly smaller callosal area than the control males. Cocaine induced no significant changes in the weight of the body or brain. There were no significant sex differences in the midline sagittal area of the anterior commissure, and no apparent effects of cocaine exposure were determined in this structure. These findings suggest that early postnatal exposure to cocaine abolishes the sexual differentiation of the corpus callosum in male rats.

Animals↗

PubMed1996

Prenatal diagnosis of abdominal wall defects and their prognosis.

To determine whether associated anatomic features may be useful in predicting fetal prognosis, 43 fetuses who had ultrasonographic evaluation for abdominal wall defects during a 13-year period were reviewed. Thirty-one fetuses had omphalocele and were classified into three groups according to fetal ultrasonography results: ruptured omphalocele with exposed liver, giant omphalocele with exposed liver, and small omphalocele without liver herniation. Twelve fetuses had gastroschisis. Ten of the 12 fetuses with gastroschisis survived; one died in utero. Nine of the 12 with a small omphalocele survived. Ten of 12 fetuses with giant omphalocele survived, but six of the seven with ruptured omphalocele died of pulmonary hypoplasia and respiratory insufficiency. Ruptured omphalocele recognized in utero was accompanied by intrauterine growth retardation and liver herniation and frequently was associated with deformity of the spine, diaphragmatic defects, vesicointestinal fissure, and meningocele. These results indicated that, together with the deformity of the spine, rupture and absence of the covering membrane with an exposed liver (noted through fetal ultrasonography) may suggest a poor prognosis because of pulmonary hypoplasia.

Abdominal Muscles↗

PubMed1996

Dominant negative effect of the truncated p110 subunit of phosphatidylinositol-3 kinase.

Effects of p110, the catalytic subunit of PI-3 kinase, on induction of TPA response element-driven promoter by EGF was examined. The induction was enhanced by co-expression of the wild type of p110. The truncated p110 mutants containing the binding site for p85 but missing the catalytic activity repressed the induction. A mutant with no binding activity to p85 did not show this effect. These results suggest that PI-3 kinase is involved in signal transduction of EGF and that the truncated p110s capable of binding to p85 serves as a dominant negative reagent for PI-3 kinase.

Animals↗

PubMed1996

Two phosphorylations specific to the tail region of the 204-kDa heavy chain isoform of porcine aorta smooth muscle myosin.

In a porcine aorta extract, we observed two protein kinase activities which specifically phosphorylate the 204-kDa heavy chain isoform of aorta myosin in the absence of conventional kinase activators. We referred to these two protein kinases, eluted at 0.15 and 0.2 M KCl from a DEAE-column, as myosin kinases I (MKI) and II (MKII), respectively. The phosphorylation site for MKI was determined using a purified phosphopeptide derived from porcine aorta myosin phosphorylated with MKI. By comparison with the deduced amino acid sequence for smooth muscle myosins, the site corresponded to a Ser located at 3 amino acids upstream from a Pro, the putative end of the alpha-helical segment of the 204-kDa heavy chain tail. A homologous Ser is only present in smooth muscle myosins, i.e. not in nonmuscle myosins. MKI was purified 130-fold, but not separated from a kinase activity phosphorylating Ser1 or Ser2 in the 20-kDa regulatory light chain of aorta myosin. In contrast, MKII was purified to near homogeneity. MKII phosphorylated the porcine aorta myosin heavy chain at a Ser 19 amino acids downstream from the MKI site. The amino acid sequence around the Ser shared a consensus sequence of the phosphorylation site. The amino acid sequence around the Ser shared a consensus sequence of the phosphorylation site for casein kinase II and was homologous to that reported for bovine aorta myosin [Kelley, C.A. and Adelstein, R.S. (1990) J Biol. Chem. 265, 17876-17882]. MKII was identified as a multifunctional protein kinase, casein kinase II.

Acid Phosphatase↗

PubMed1996

Identification of interleukin-6 as a factor that induces neurite outgrowth by PC12 cells primed with NGF.

We have screened for a factor that induces neurite outgrowth by PC12 cells only after NGF pretreatment in the supernatants of 38 transformed cell lines, finding three positive clones. The factor showing the strongest activity was purified and identified as IL6. In this NGF-IL6 system, the process of neurite outgrowth by PC12 cells can be separated into NGF-dependent and IL6-dependent steps. The IL6-dependent step requires RNA synthesis, suggesting that IL6 induces new gene expression depending on NGF-priming. These findings suggest that the gene expression during the differentiation process is regulated by at least two signals.

Animals↗

PubMed1996

Molecular and biochemical characterization of three anthocyanin synthetic enzymes from Gentiana triflora.

Full length cDNA clones of flavonoid 3',5'-hydroxylase, dihydroflavonol 4-reductase and flavonoid 3-glucosyltransferase were cloned from petals of Gentiana triflora. Their sequences were homologous to counterparts from other plants. Flavonoid 3',5'-hydroxylase and flavonoid 3-glucosyltransferase were enzymatically characterized by expressing cDNAs in heterologous expression systems.

Alcohol Oxidoreductases↗

PubMed1996

Synergistic effect of alanine and glycine on bovine embryos cultured in a chemically defined medium and amino acid uptake by vitro-produced bovine morulae and blastocysts.

The effect of glycine and alanine on the development of 2- to 4-cell bovine embryos, and amino acid uptake by bovine morulae and blastocysts, were examined through the use of a chemically defined medium. Bovine embryos at 2- to 4-cell stages were prepared by in vitro maturation and fertilization and cultured in a synthetic oviduct fluid medium (SOFM) containing polyvinyl alcohol (PVA) instead of BSA in order to examine the effect of amino acids. Morulae or blastocysts obtained from culture in SOFM containing BSA were cultured for 10 h in SOFM containing PVA to determine amino acid uptake. The combination of essential and nonessential amino acids with or without glutamine improved development to the blastocyst stage over that observed in the control (34% or 32% vs. 19%, respectively; p < 0.05). The optimal supplemental concentrations were 5 mM for alanine and 10 mM for glycine. At these concentrations, development to blastocysts was enhanced by the addition of alanine or glycine independently (35% or 36% vs. 26%, respectively; p < 0.05); the combined addition of alanine and glycine greatly (p < 0.01) improved proportions of blastocysts (45% vs. 26%) and hatched blastocysts (10% vs. 2%) compared to those obtained in the control. Addition of glycine with or without alanine to culture medium significantly increased cell number per blastocyst over that in the control (137 +/- 5 or 131 +/- 5 vs. 106 +/- 4, respectively; p < 0.01). Bovine morulae and blastocysts depleted aspartate, serine, and glutamate at a highly significant rate (p < 0.001) and arginine at a significant rate (p < 0.05), and produced alanine at a highly significant rate (p < 0.001) when cultured in medium containing 20 essential and nonessential amino acids. Serine, asparagine, glycine, alanine, and glutamine were highly (p < 0.001) produced by bovine morulae and blastocysts cultured in a medium containing essential amino acids without glutamine. These results indicate that alanine and glycine in a defined medium synergistically improve development of in vitro-produced bovine embryos, and also that bovine morulae and blastocysts prefer aspartate, glutamate, serine, and arginine and produce glutamine and several nonessential amino acids (serine, asparagine, alanine, and glycine).

Alanine↗

PubMed1996

Hepatic saturation mechanism of ethanol: application of mathematical models to ethanol outflow profiles in the perfused rat liver.

The saturation mechanism of hepatic ethanol (EtOH) elimination was studied in the perfused rat liver. EtOH outflow profiles after the instantaneous administration of 3 (mg/ml) x 0.4(ml), 12 x 0.1, 24 x 0.1, and 3 x 0.1 mg (as a dose concentration x a volume) through the portal vein were analyzed by the statistical moment analysis and mathematical models (i.e., dispersion models). Results for 3 x 0.1 and 12 x 0.1 mg doses by moment analysis were similar. This demonstrated that the elimination exhibits linear kinetics. Recovery ratio and hepatic volume of distribution for 3 x 0.4 and 24 x 0.1 mg were larger than those for 3 x 0.1 and 12 x 0.1 mg doses and were similar. Kinetics after administration of 3 x 0.4 and 24 x 0.1 mg may be nonlinear. A difference in the relative dispersion (CV2) obtained by moment analysis between 3 x 0.4 and 24 x 0.1 mg doses indicated different properties of the nonlinear elimination kinetics. There were no differences in all the parameters in the one-compartment dispersion model between 3 x 0.4 and 24 x 0.1 mg doses. In the two-compartment dispersion model, there were differences in the blood volume (VB) and the forward partition rate constant (k12) between 3 x 0.4 and 24 x 0.1 mg (p < 0.05), whereas the elimination rate constant (k sigma) and the dispersion number values for these doses were similar. These findings demonstrated that there is difference in the no-equilibrium process between 3 x 0.4 and 24 x 0.1 mg doses. Therefore, we suggest that the continuous EtOH input into the liver causes the saturation of enzyme pathways and the change of the nonequilibrium process.

Animals↗

PubMed1996

Evidence that cytochrome P-4502E1 contributes to ethanol elimination at low doses: effects of diallyl sulfide and 4-methyl pyrazole on ethanol elimination in the perfused rat liver.

The roles of cytochrome P-4502E1 and alcohol dehydrogenase (ADH) on ethanol (EtOH) hepatic elimination was examined in the perfused rat liver. EtOH concentration-time curves of outflow after instantaneous administration (0.46 mg) through the portal vein with or without perfusion of diallyl sulfide (DAS), a selective cytochrome P-450E1 inhibitor, and/or 4-methyl pyrazole (4-MP), a classical ADH inhibitor, were analyzed by the statistical moment analysis and the compartment dispersion model. Recovery ratios obtained by moment analysis significantly changed with perfusion of inhibitors (p < 0.01). Values of the hepatic volume of distribution and the relative dispersion were significantly higher by the perfusion of DAS and 4-MP (p < 0.01). In the two-compartment dispersion model, the partition ratio (K') and the first-order elimination constant (K0) were decreased significantly by DAS (p < 0.05). By the addition of 4-MP, the blood volume of distribution (VB) and the backward partition rate constant (k21) were increased significantly (p < 0.05). K sigma values were decreased significantly to 0 (p < 0.001). The decrease of elimination rates by DAS and/or 4-MP shows the inhibition of metabolic pathways. The change of V beta and k21 caused by DAS and 4-MP indicates that EtOH taken into hepatic tissues was not metabolized and flowed out into the perfusates. Inhibition rates calculated from the efficiency number with addition of DAS and DAS + 4-MP were 40.7 and 99.3%. Therefore, cytochrome P-4502E1 and ADH accounted for 40 and 60% of the hepatic EtOH elimination at low doses.

Alcohol Dehydrogenase↗

PubMed1996

Role of chaos in trial-and-error problem solving by an artificial neural network.

One role of chaotic neural activity is illustrated by means of computer simulations of an imaginary agent's goal-oriented behavior. The agent has a simplified neural network with seven neurons and three legs. The neuronal network consists of one photosensory neuron and three pairs of inter- and motor neurons. The three legs whose movements are governed by the three motor neurons allow the agent to walk in six concentric radial directions on a plane. It is intended that the neural network causes the agent to walk in a direction of greater brightness, to reach finally the most brightly lit place on the plane. The presence of only one sensory neuron has an important meaning. That is, no immediate information on directions of greater brightness is sensed by the agent. In other words, random walking in the manner of trial- and error problem solving must be involved in the agent's walking. Chaotic firing of the motor neurons is intended to play a crucial role in generating the random walking. Brief random walking and rapid straight walking in a direction of greater brightness were observed to occur alternately in the computer simulation. Controlled chaos in naturally occurring neural networks may play a similar role.

Animals↗

PubMed1996

A novel differentiation factor for PC12 cells from culture supernatant of mouse hepatocyte cell line MLE-15A2.

We have found a factor that induces neurite outgrowth of rat PC12 cells in the culture supernatant of the cell line MLE-15A2. This factor was designated as MDDF. The factor was sensitive to protease, dithiothreitol, and high-temperature treatments. The apparent molecular mass was 80 kDa on Superdex 200 gel filtration. No significant tyrosine phosphorylation was detected after MDDF stimulation in Western blotting analysis with anti-phosphotyrosine antibody, suggesting that the signal transduction may not be mediated by a tyrosine kinase cascade that is involved in signaling of most of the known factors. Activation of MAP kinase was very weak and was seen only 5 min after stimulation, suggesting that prolonged activation of MAP kinase was not required for neurite outgrowth induced by MDDF. Because the biochemical characteristics of MDDF are different from those of any known peptide factors that induce neurite outgrowth of PC12 cells, MDDF may be a novel differentiation factor for PC12 cells.

Animals↗

PubMed1996

Effect of medium renewal during culture in two different culture systems on development to blastocysts from in vitro produced early bovine embryos.

Three experiments investigated the effects of culture systems and renewal of medium (synthetic oviduct fluid medium (SOFM) supplemented with glutamine and amino acids) during in vitro development of in vitro-produced 2- to 4-cell bovine embryos. In Exp. 1, the early cleaved embryos were cultured under three different conditions for 186 h using 24-well dishes. The proportions of blastocysts (P < .05) and hatched blastocysts (P < .001) were lower for culture in SOFM + .8% BSA with renewal of medium at 48-h intervals (26.7 and 7.9%) than for cultures in SOFM + 10% human serum without renewal of medium (32.6 and 20.6%) and in SOFM + BSA without renewal of medium (32.2 and 23.0%). Two culture systems (well and microdrop) with or without renewal of medium (SOFM + BSA) were compared in Exp. 2 (186-h culture) and 3 (138-h culture). In Exp. 2, the microdrop system was superior (P < .05) to the well system for development of hatched blastocysts (19.1 and 11.7%), and the highest development to blastocysts (39.6%) and hatched blastocysts (27.8%) was observed in the microdrop system without renewal of medium. Experiment 3 also showed that the microdrop system was superior (P < .05) to the well system for development to blastocysts (32.7 and 27.2%), and the highest development rate to blastocysts (37.6%) occurred without renewal of medium. When the medium was not renewed in Exp. 2 and 3, the ammonium concentrations increased in both the well and microdrop systems. The results indicate that despite the reduction in ammonium concentrations resulting from medium renewal, renewal resulted in reduced development of embryos to hatched blastocysts in both the well and microdrop systems.

Amino Acids↗

PubMed1996

Death caused by undiagnosed acute pancreatitis.

A fatal case of acute pancreatitis is reported. On account of difficulty in eating caused by persistent dysphagia and heartburn, the clinical condition of a 39-year-old man who had been a heavy drinker deteriorated rapidly. He was taken to a hospital in an ambulance in an unconscious state. Based on the endoscopic examination and blood chemistry data, the diagnosis of hemorrhagic esophagitis and hepatic failure was made. Treatment including fluid infusion was unsuccessful and he died on the second hospital day. Based on a strong suspicion that the pathologic change in the esophagus may have been chemical esophagitis caused by corrosives of some type, the police ordered an administrative autopsy. The postmortem examination revealed marked necrosis in the pancreas and in the abdominal fatty tissue including the omentum and the mesentery. The necrotic areas in the pancreas were accompanied by only a slight degree of hemorrhage. The cause of death was diagnosed as acute pancreatitis. The pathologic change in the esophagus was identified as Candida esophagitis. Alcohol abuse and malnutrition caused by esophagitis were both considered to be factors which lead to the acute fatal pancreatitis.

Acute Disease↗