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Biomedical subjects

Y Fukui

Publications and source records attributed to Y Fukui.

At least 307 records · Page 17Linked to original sources

Indocyanine green test is a reliable indicator of postoperative liver function in biliary atresia.

In order to evaluate liver function in postoperative biliary atresia (BA) patients, the indocyanine green (ICG) clearance test was performed. Nineteen patients, ranging in age from 2 to 9 years, were classified into three groups. Group I were those who had no jaundice throughout the postoperative period (n = 9), Group II were those who had recurrent or intermittent jaundice (n = 7), and Group III were those who had persistent jaundice after hepatic portojejunostomy (n = 3). Forty-four volunteers of comparative age and sex served as controls to establish a normal range of ICG-K values in infants and children. The normal minimal ICG-K value in those 2-9 years of age was defined as 0.19. ICG-K values in BA patients were 0.262 +/- 0.073 (Group I), 0.150 +/- 0.060 (Group II), and 0.054 +/- 0.011 (Group III). All nine patients with ICG-K below the normal level had esophageal varices, and seven of nine had a history of gastrointestinal hemorrhage. Two patients with ICG-K values within the normal range had esophageal varices, and none had a history of gastrointestinal hemorrhage. Five of nine patients with ICG-K values below normal had low serum albumin levels, and all had low prealbumin levels. Of those with ICG-K within the normal range, none had low serum albumin or prealbumin levels. On the other hand, serum bilirubin and other liver function tests did not accurately reflect clinical findings. In conclusion, the ICG test using the ICG-K value was confirmed to be a reliable indicator of postoperative liver function in BA patients.

Biliary Atresia↗

Structure of 85 kDa subunit of human phosphatidylinositol 3-kinase analyzed by using monoclonal antibodies.

An 85 kDa subunit (p85 alpha) of phosphatidylinositol 3-kinase (PI-3K) has one SH3 and two SH2 regions [SH2(N) and SH2(C)], which direct protein-protein interaction. We have established eighteen hybridomas producing monoclonal antibodies against p85 alpha to study the structure-function relationship of this protein. Epitope mapping using a series of deletion mutants expressed in E. coli showed that the monoclonal antibodies bound to at least 5 distinct epitope regions, which were well dispersed on p85 alpha except for its carboxyl-terminus. Monoclonal antibodies against amino-terminal regions and polyclonal antibodies against carboxyl-terminal regions immunoprecipitated p85 alpha expressed in human cells and in E. coli. On the other hand, monoclonal antibodies against the central part of p85 alpha failed to immunoprecipitate p85 alpha efficiently; however, they could immunoprecipitate p85 alpha mutants with deletion of either the amino- or the carboxyl-terminal region. Similar results were obtained by immunocytochemistry using confocal microscopy. These results suggested that steric hindrance prevents binding of monoclonal antibodies to the central part of p85 alpha where SH2(N) is located. The SH2(N) may have a distinct function from SH2(C), which is located at the carboxyl-terminal region and has been shown to mediate the binding of PI-3K to activated growth factor receptors.

Animals↗

Development of an all-in-one percutaneous cardiopulmonary support system.

This paper deals with development of an all-in-one percutaneous cardiopulmonary support (PCPS) system. In recent years, PCPS has been used for the treatment of acute myocardial infarction. A prototype of a compact all-in-one PCPS system was developed. The system contains a centrifugal pump and an extra-capillary flow-type membrane lung in one body. The system has a priming volume of 250 ml, which allows for PCPS with no additional blood. The in vitro tests and an ex vivo test were conducted. The system produces 1.6-5 L/min of flow in the experiments. The O2 transfer rate was 310 ml/min, and the CO2 transfer rate was 300 ml/min at a blood flow rate of 5 L/min. This device is compact, requires less priming volume than a standard system, and is easy-to-handle in the experiments. The system is considered applicable to percutaneous cardiopulmonary support.

Animals↗

Both the SH2 and SH3 domains of human CRK protein are required for neuronal differentiation of PC12 cells.

Human CRK protein is a homolog of the chicken v-crk oncogene product and consists mostly of src homology region 2 (SH2) and SH3, which are shared by many proteins, in particular those involved in signal transduction. SH2 has been shown to bind specifically to phosphotyrosine-containing peptides. We report here that both SH2 and SH3 are required for signaling from CRK protein. Microinjection of the CRK protein induced neurite formation of rat pheochromocytoma cell line PC12. This activity was abolished by mutation of the CRK protein in either SH2 or SH3. The neuronal differentiation induced by the CRK protein was blocked by an excess amount of peptides containing CRK SH3. Moreover, we identified three proteins, of 118, 125, and 136 kDa, which bound specifically to CRK SH3. The CRK-induced neuronal differentiation was also suppressed by monoclonal antibodies against either CRK SH2 or p21ras. These results suggest that both SH2 and SH3 of the CRK protein mediate specific protein-protein binding and that the resulting multimolecular complex generates a signal for neurite differentiation through activation of p21ras.

Amino Acid Sequence↗

Protothecosis in a patient with systemic lupus erythematosus.

A rare case of protothecosis in a 40-year-old Japanese woman with systemic lupus erythematosus, under long-term prednisolone treatment is presented. The patient developed extensive ulcerative skin lesions after injury. The characteristic histopathological findings and biochemical sugar assimilation tests indicated infection by Prototheca wicherhamii. The lesions improved following treatment with amphotericin B. The immunodeficient state of this patient seemed to contribute to the infection with Prototheca.

Abscess↗

Effects of methylmercury on the brain of infant rats reared artificially.

Fetal Minamata Disease leads to cerebral palsy, blindness, and deafness accompanying microcephaly. Fetal Minamata Disease has been considered to be exposed to methylmercury during mid and late gestation periods. Brain development in the late gestation in human seems to be equivalent to that in the postnatal stage in rats and mice. Artificial rearing systems for infant rats exclude the maternal and nutrient factors. We administered methylmercury to the infant rats in the artificial rearing system attempting to produce an animal model for Fetal Minamata Disease. Brain weight and brain weight/body weight ratio decreased, and pyknotic degeneration of the nerve cells in the inner granular layer of the cerebellum increased in the groups treated with methylmercury. But, we could not observe drastic degeneration in their brains which mimicked Fetal Minamata Disease in human.

Animals↗

[An experimental and clinical study of peripheral nerve potentials evoked by stimulating the spinal cord].

The purpose of this study was to evaluate a possible use of TNP (tibial nerve potentials evoked by stimulating the upper thoracic spinal cord) as waveforms for spinal cord monitoring. The results of an experimental study in cats indicate that TNP mainly conducts antidromically in the dorsal column of the spinal cord with a velocity of approximately 70 m/sec. TNP was not a motor-evoked potential, but it involved some potential conducted from the ventral root, which was very small in amplitude. In men, TNP consisted of several negative deflections of 20 msec in onset latency and 1 microV in amplitude. The peak latency and amplitude of TNP were used as parameters for monitoring. Using this method 40 cases with spinal disorders were monitored. During surgery, the peak amplitude of TNP fluctuated within a range of 20% of the control wave in 24 cases, increased in three cases, and decreased in six cases. There was no false negative monitoring. TNP could not be recorded in four cases, in which SC-DESCP was used for monitoring instead of TNP. TNP disappeared during operation in three cases and in one of the cases, the postoperative neurological status had deteriorated. The disappearance of TNP was considered to be a positive sign of postoperative neurological complications. This study has concluded that TNP can be used for spinal cord monitoring, especially in patients with thoracolumbar disorders.

Adolescent↗

Insulin-dependent formation of a complex containing an 85-kDa subunit of phosphatidylinositol 3-kinase and tyrosine-phosphorylated insulin receptor substrate 1.

Monoclonal antibodies raised against the 85-kDa subunit (p85) of bovine phosphatidylinositol (PI) 3-kinase were found to recognize uncomplexed p85 or p85 in the active PI 3-kinase. Immunoprecipitation studies of Chinese hamster ovary cells, which overexpress the human insulin receptor when treated with insulin, showed increased amounts of p85 and PI 3-kinase activity immunoprecipitable with monoclonal anti-p85 antibody and no increase in the tyrosine phosphorylation of p85. Insulin also induced an association of p85 with the tyrosine-phosphorylated insulin receptor substrate 1 (IRS-1) and other phosphorylated proteins ranging in size from 100 to 170 kDa but not with the activated insulin receptor. In vitro reconstitution studies were used to show p85 in the active PI 3-kinase associated with the tyrosine-phosphorylated IRS-1 but not with the activated insulin receptor. Competition studies using synthetic phosphopeptides corresponding to potential tyrosine phosphorylation sites of IRS-1 revealed that phosphopeptides containing YMXM motifs inhibited this association with different potencies, whereas nonphosphorylated analogues and a phosphopeptide containing the EYYE motif had no effect. Src homology region 2 domains of p85 expressed as glutathione S-transferase fusion proteins also bound to tyrosine-phosphorylated IRS-1. These results suggest that insulin causes the association of PI 3-kinase with IRS-1 via phosphorylated YMXM motifs of IRS-1 and Src homology region 2 domains of p85.

Amino Acid Sequence↗

Potent and specific inhibition of p60v-src protein kinase both in vivo and in vitro by radicicol.

A fungal metabolite, radicicol, with a macrocyclic ring induced the reversal of transformed phenotypes of v-src-transformed fibroblasts (Rous sarcoma virus-transformed 3Y1 rat fibroblast) at a quite low concentration of 0.1 microgram/ml. Actin stress fibers reappeared in the transformed cells after treatment with radicicol. Radicicol reduced the intracellular level of autophosphorylation of p60v-src as well as the level of other tyrosine-phosphorylated proteins in a dose-dependent manner. In vitro kinase assay revealed that radicicol effectively inhibited not only autophosphorylation but also transphosphorylation activities of purified p60v-src with a concentration producing 50% inhibition of 0.1 microgram/ml. However, radicicol showed no inhibitory effect on protein kinase C or protein kinase A. These results suggest that radicicol is a novel and specific protein-tyrosine kinase inhibitor and that the decreased level of tyrosine kinase activity of p60v-src causes reversion of transformed phenotypes of Rous sarcoma virus-transformed 3Y1 rat fibroblast. Furthermore, differentiation of Friend leukemia cells, which is one of the known characteristic phenomena associated with the inhibition of tyrosine kinase, was also induced in the concentration range of 0.05-0.5 microgram/ml, suggesting that the agent is useful for the analysis of differentiation as well as the kinase-mediated signal transduction.

Animals↗

Osteoclasts express high levels of p60c-src, preferentially on ruffled border membranes.

Expression of p60c-src, the normal cellular counterpart of the transforming protein of Rous sarcoma virus (RSV), p60v-src, was examined in mouse and rat authentic osteoclasts and mouse osteoclast-like multinucleated cells (MNCs) formed in vitro. In co-cultures of mouse osteoblastic cells and spleen cells, the expression of p60c-src strikingly increased on day 5 in parallel with the appearance of MNCs in the presence of 1 alpha,25-dihydroxyvitamin D3 (1 alpha,25(OH)2D3). Immunohistochemical examination confirmed the high level expression of p60c-src in both mouse authentic osteoclasts and MNCs. Electron microscopic examination revealed that p60c-src was primarily localized on ruffled border membranes and vacuoles, but not on the clear zone in rat authentic osteoclasts. These results suggest that p60c-src is important in osteoclastic bone resorption.

Animals↗

Insulin treatment stimulates the tyrosine phosphorylation of the alpha-type 85-kDa subunit of phosphatidylinositol 3-kinase in vivo.

After adding insulin to cells overexpressing the insulin receptor, the activity of phosphatidylinositol (PI) 3-kinase in the anti-phosphotyrosine immunoprecipitates was rapidly and greatly increased. This enzyme may therefore be a substrate for the insulin receptor tyrosine kinase and may be one of the mediators of insulin signal transduction. However, it is unclear whether or not activated tyrosine kinase of the insulin receptor directly phosphorylates PI 3-kinase at tyrosine residue(s) and whether insulin stimulates the specific activity of PI 3-kinase. We reported previously that the 85-kDa subunit of purified PI 3-kinase was phosphorylated at tyrosine residue(s) by the insulin receptor in vitro. To examine the tyrosine phosphorylation of PI 3-kinase and change of its activity by insulin treatment in vivo, we used a specific antibody to the 85-kDa subunit of PI 3-kinase. The activity of PI 3-kinase in immunoprecipitates with the antibody against the p85 subunit of PI 3-kinase was increased about 3-fold by insulin treatment of cells overexpressing insulin receptors. Insulin treatment also stimulated the tyrosine, serine, and threonine phosphorylation of the alpha-type 85-kDa subunit of PI 3-kinase in vivo. Phosphatase treatment of the immunoprecipitates abolished the increase in PI 3-kinase activity. The phosphorylation(s) of the kinase itself, tyrosine phosphorylation(s) of associated protein(s), or the complex formation of the phosphorylated PI 3-kinase with associated proteins may increase the activity of PI 3-kinase.

Animals↗

Activation of Src-like protein-tyrosine kinase Lyn and its association with phosphatidylinositol 3-kinase upon B-cell antigen receptor-mediated signaling.

Crosslinking of membrane-bound immunoglobulins, which are B-cell antigen receptors, causes proliferation and differentiation of B cells or the inhibition of their growth. The receptor-mediated signaling involves tyrosine phosphorylation of cellular proteins. The Src-like protein-tyrosine kinase Lyn is expressed preferentially in B cells and is an intracytoplasmic constituent of the B-cell antigen receptor complex. Crosslinking of membrane-bound immunoglobulin M with antibody induced rapid increases in the kinase activities of Lyn and Lyn-associated phosphatidylinositol 3-kinase. Crosslinking of B-cell antigen receptor also induced association of Lyn with an 85-kDa noncatalytic subunit of phosphatidylinositol 3-kinase. Thus, Lyn is functionally associated with membrane-bound immunoglobulin M and seems likely to participate in B-cell antigen receptor-mediated signaling.

B-Lymphocytes↗

Parthenogenetic development of bovine oocytes treated with ethanol and cytochalasin B after in vitro maturation.

The present study was conducted to investigate the effects of different culture durations (24-36 hr) on bovine oocyte maturation in vitro and the effect of the presence or absence of cumulus cells at the time of treatment to induce parthenogenetic activation (exposure to ethanol and cytochalasin B; CB) (experiment I). The effects of dosage (2.5 or 5.0 micrograms/ml) and incubation time (2.5, 5, or 10 hr) in CB (experiment II) on the subsequent development to the blastocyst stage in vitro was also investigated. In experiment I, cleavage and development to the blastocyst stage were not affected by the presence or absence of cumulus cells at the time of parthenogenetic activation. However, the 24-hr culture duration for in vitro maturation had a significantly lower rate of development to the blastocyst stage than the longer culture durations (27-36 hr). In experiment II, treatment with 5 micrograms/ml CB for 5 hr showed the highest percentage of development to blastocyst in the oocytes matured for both 27 and 30 hr. To determine the viability of the parthenogenetic embryos (morulae and blastocysts), four recipient heifers received two embryos each, and one heifer was found to be pregnant on day 35 following transfer. Although fetal heartbeat was not observed, the subsequent estrus was prolonged in all heifers. The present results demonstrate development of in vitro-matured, parthenogenetically activated bovine embryos up to the preimplantation stage.

Animals↗

Cardiovascular simulation using a multiple modeling method on a digital computer--simulation of interaction between the cardiovascular system and angiotensin II.

A cardiovascular system model that simulates interactive responses to drugs has been developed on a small digital computer. The overall model basically consists of three models. The first is a momentum transport model that represents relations between blood pressure and flow in the cardiovascular system. In this model, the cardiovascular system is divided into 14 components and modeled by using equivalent electrical circuits. The second is a mass transport model comprising 14 compartments corresponding to the respective components of the cardiovascular system. This model represents the distribution of the administered drug in the various cardiovascular components. The third is an interaction model that represents the relationships between the momentum and mass transport models. This model causes variations in the resistance and capacitance parameters of the momentum transport model as a function of the current drug concentrations in the appropriate compartments of the mass transport model. The capacitances representing the ventricles are varied in a time-dependent fashion to simulate the beat of the heart. Simulation is performed by using the Euler method to solve a system of 28 ordinary differential equations governing the momentum and mass transport models on a 32-bit microcomputer, a Macintosh II. The model was assessed by performing two demonstrations of the cardiovascular response to the vasopressor angiotensin II (AT II). They first examined the interaction between the cardiovascular system and AT II. The effect of AT II on the cardiovascular system was incorporated into the interaction model. Administration of AT II as a constant infusion (200 micrograms/hr) resulted in an elevation of mean arterial pressure from approximately 100 to 150 mm Hg.(ABSTRACT TRUNCATED AT 250 WORDS)

Angiotensin II↗

Heart rate and body temperature sensitive diaphragm pacing.

Two ways of rate control for diaphragm pacing are proposed. One is rate control using only the patients' body temperature (method I). The other is rate control by both the patients' heart rate and body temperature (method II). To test the effectiveness of these methods, a diaphragm pacemaker which can be controlled by both heart rate and body temperature has been developed. It was applied to nine mongrel dogs. The pacing rate is controlled by atrial blood temperature (method I) or by both heart rate and temperature (method II). The animal's metabolism was elevated by the administration of a pyrogenic drug. It was found that method I is not suited to rapid changes in metabolism; however, it is useful in extreme metabolic elevation. An animal's metabolism was supported by using method II in all ranges of metabolism. This method proved more effective than method I for rate-responsive diaphragm pacing.

Animals↗

Intrapancreatic duodenal duplication associated with pancreatic pseudocysts.

A rare case of intrapancreatic duodenal duplication causing pancreatitis is reported. At 2 years of age, the patient presented with a recurrent pancreatic pseudocyst. Intraoperative pancreatogram showed the presence of cystic duodenal duplication in the aberrant lobe of the pancreas communication with the pancreatic duct. Since the resection of the duplication, she has been free from recurrence of pancreatitis. In this case, intraoperative pancreatography was of great value.

Duodenum↗

The superior cervical ganglion: origin of sympathetic fibers in the facial and hypoglossal nerves in the cat.

Location of superior cervical ganglion (SCG) neurons, sending axons into the facial and hypoglossal nerves, was investigated in the cat by means of retrograde axonal transport of horseradish peroxidase (HRP). After wheat germ agglutinin conjugated HRP (WGA-HRP) was injected into these nerves, many retrogradely labeled neurons were found widely in the ipsilateral SCG, particularly around the caudal half of the SCG. These neurons were round or oval in shape and 70-80% of these were medium in size. In fluorescent experiments, fast blue (FB) was used in combination with diamidino yellow (DY). After injections of FB into the facial nerve and DY into the hypoglossal nerve, a few FB-DY double-labeled neurons occurred in the SCG ipsilaterally.

Adrenergic Fibers↗