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Biomedical subjects

K Abe

Publications and source records attributed to K Abe.

At least 271 records · Page 15Linked to original sources

Absence of aflatoxin biosynthesis in koji mold (Aspergillus sojae).

Ten strains isolated from industrial soy sauce producing koji mold were identified as Aspergillus sojae and distinguished from Aspergillus parasiticus morphologically and physiologically. There was no detectable aflatoxin in any culture extracts of A. sojae strains. Strain 477 was chosen as a representative strain of industrial A. sojae for further molecular analysis. All enzymatic activities associated with the aflatoxin biosynthesis were not detected or negligible in strain 477 compared with that of the A. parasiticus strain. Southern analysis suggested that the genomic DNA of strain 477 contained aflatoxin biosynthetic pathway genes. In contrast, all industrial strains lacked detectable transcripts of aflR, the main regulatory gene for aflatoxin biosynthesis, under the aflatoxin-inducing condition. Our data suggest that defects in aflR expression cause the lack of expression of aflatoxin-related genes which results in the absence of aflatoxin biosynthesis in A. sojae strains.

Aflatoxins↗

Pre-termination in aflR of Aspergillus sojae inhibits aflatoxin biosynthesis.

The aflR gene product is the main transcriptional regulator of aflatoxin biosynthesis in Aspergillus parasiticus and Aspergillus flavus. Although A. sojae strains do not produce aflatoxins, they do have an aflR homologue. When compared with the aflR of A. parasiticus, the A. sojae gene contains two mutations: an HAHA motif and a premature stop codon. To investigate the functionality of the A. sojae aflR gene product, we used a GAL4 one-hybrid system in yeast. The transcription-activating activity of AflR from A. sojae was 15% of that from A. parasiticus. The introduction of an additional aflR from A. sojae into an A. parasiticus strain did not affect aflatoxin productivity. A hybrid aflR comprising the amino-terminal region of A. sojae aflR and the carboxy-terminal region of A. parasiticus aflR suppressed the effect associated with pre-termination of the A. sojae AflR. We conclude that the premature stop codon of the A. sojae aflR is the key to its functionality and leads to prevention of aflatoxin biosynthesis through loss of the transcription of aflatoxin biosynthesis-related genes.

Aflatoxins↗

Venous hemangioma of the mediastinum.

Mediastinal hemangiomas are rare tumors. We present a case of venous hemangioma of the mediastinum. Computed tomography shows anterior mediastinal solid mass without enhancement effects. We believe that it is necessary to include a mediastinal hemangioma for differential diagnosis, when little or no enhancement effects are noted at early and delayed CT examination.

Adult↗

Molecular cloning, genetic mapping, and expression of the mouse Sf3b1 (SAP155) gene for the U2 snRNP component of spliceosome.

SAP155, a subunit of the U2 snRNP, is essential for prespliceosome assembly and splicing catalysis of the major spliceosome. Moreover, the protein has been identified in the minor (U12-dependent) spliceosome. These facts strongly suggest that SAP155 is shared by two distinct complexes owing to its importance in the removal of any type of intron. Here we have isolated a cDNA encoding the 146-kDa mouse homolog, designated Sf3b1. The amino acid sequence of Sf3b1 is very highly conserved among homologs from Schizosaccharomyces pombe (52.4% identity) to human (99.6%), and the C-terminal 825 residues of these Sf3b1 homologs show even higher identities. This C-terminal region shows significant similarity to the PR65 subunit of protein phosphatase 2A, which is composed of 15 tandem repeats of a 39 amino acid sequence. Mouse genome analyses showed Sf3bh1 to be a single-copy gene mapping to the central part of Chromosome (Chr) 1. Northern blot analysis and whole mount in situ hybridization revealed Sf3b1 to be ubiquitously expressed in a variety of adult tissues and mid-gestation embryos.

Amino Acid Sequence↗

Thinning and small holes at an impending tear of a retinal pigment epithelial detachment.

BACKGROUND: A tear of a retinal pigment epithelial detachment (PED) suddenly exposes a large area of bare Bruch's membrane. We report here the case of a patient whom we observed during the gradual, spontaneous development of a PED tear. METHOD: A 5.25-year case study of a 67-year-old woman with bilateral serous PEDs. RESULTS: Retinal pigment epithelial (RPE) thinning or small holes were seen along the PED margin in both eyes. Fluorescein angiograms showed intense hyperfluorescence without leakage, and indocyanine green angiography showed choroidal vessels through regions of RPE thinning or small holes. Optical coherence tomographs showed an interruption of a hyperreflective band corresponding to retinal pigment epithelium. A typical tear of the PED ensued later. CONCLUSION: Multiple, small regions of RPE thinning or holes along the margin of PED can be a sign of an impending PED tear.

Aged↗

Ophthalmological findings in patients with spinocerebellar ataxia type 1 are not correlated with neurological anticipation.

BACKGROUND: Optic atrophy, attenuation of the oscillatory potentials (OPs) of the electroretinogram (ERG), and enlargement of corneal endothelial cells, have been reported in patients with spinocerebellar ataxia type 1 (SCA1). These patients have a trinucleotide repeat expansion in the SCA1 gene and show neurological anticipation. The purpose of this study was to determine whether the ophthalmological findings are correlated with the neurological disorders, and whether ophthalmological anticipation is present in patients with SCA1. METHODS: The visual acuity, ERGs, and corneal endothelial cell density were examined in 14 patients whose DNA analysis revealed an expanded trinucleotide repeat in an allele of the SCA1 gene. The results of the tests were compared with the trinucleotide repeat number and the duration of the neuronal disease. RESULTS: The neurological disorders in the patients showed anticipation. The negative correlation between the trinucleotide repeat number and the neurological disorder was statistically significant (P<0.0001). However, the correlations between trinucleotide repeat number and visual acuity, amplitude of OPs, and corneal endothelial cell density were not significant. Statistically significant correlations were found between the duration of the neuronal disease and the visual acuity, OPs, and corneal endothelial cell density (P<0.0001, P=0.0004, and P<0.0001, respectively). The ophthalmological disorders were prominent in patients who had neuronal disease for more than 10 years. CONCLUSION: Unlike the neurological findings, the ophthalmological disorders in patients with SCA1 were not correlated with the trinucleotide repeat number of the SCA1 gene. The ophthalmological findings were most highly correlated with the duration of the neuronal disease.

Adult↗

Sialogogic activity in the rat of peptides analogous to [Tyr8]-substance P in which substitutions have been made in the N-terminal amino acids.

In order to elucidate the regulatory roles for salivation of amino acids in positions 1-4 of the N-terminal region of [Tyr8]-substance P (SP), the structure-sialogogic activity correlations of various synthetic octa- to undecapeptides replaced in positions 1-4 of [Tyr8]-SP with each of 19 common amino acids, one by one, and with the same sequence of the C-terminal hepatapeptide as that of [Tyr8]-SP, were studied in the submandibular glands of rats after intraperitoneal injection. Each of 19 octa-, nona-, deca- and undecapeptides with replaced amino acids and a penta- to decapeptide with the progressive elimination of the N-terminal portion were newly synthesized by the multipin peptide method. All octa- to undecapeptides replaced with each of 19 common amino acids in positions 1-4 had sialogogic activities. In 19 octa- and decapeptides in which P4 and P2 had been replaced, four and three replacements, respectively, had significantly increased secretory activities. In contrast, in 19 nonapeptides in which K3 had been replaced, none had significantly increased secretory activities. Furthermore, in 19 undecapeptides in which R1 had been replaced, most replacements had significantly increased or equipotent activities for fluid secretion. It is concluded that amino acids in the N-terminal region of various tachykinins may not need to be strictly conserved and that amino acid residues in the N-terminal portion, R1 in particular and P2, may strongly inhibit secretory activity.

Amino Acid Substitution↗

Expression and postnatal changes of adrenergic receptor subtype mRNA in rat submandibular glands.

Adrenergic receptors (ARs) are involved in regulating saliva secretion and composition in salivary glands. Nine AR subtypes, including three alpha1-ARs (alpha1a-, alpha1b- and alpha1d-ARs), three alpha2-ARs (alpha2A-, alpha2B- and alpha2C-ARs) and three beta-ARs (beta1,beta2- and beta3-ARs), have been identified through molecular cloning. The five subtype genes, alpha1a-, alpha1b-, alpha2A-, beta1-, and beta2-ARs, were expressed in rat submandibular glands. In contrast, the other four subtype mRNAs, alpha1d-, alpha2B-, alpha2C- and beta3-ARs, were not detected by reverse transcription-polymerase chain reaction (RT-PCR). The steady-state mRNA expression for the five AR subtypes in rat submandibular glands was measured by quantitative competitive RT-PCR using synthetic DNA as internal standard at different stages of postnatal development. The relative rank order of AR subtype mRNA expression was alpha1a>beta2>beta1>alpha2A>alpha1b at all stages except that beta1- and alpha2A-subtypes were reversed at 2 weeks of age. The gene expression of alpha1a-AR subtype relative to total AR was low at 2 weeks of age and increased and reached a maximum at 6 weeks of age, whereas those patterns of alpha2A-, beta1- and beta2-AR subtypes were similar to each other and their gene expressions were high at 2 weeks of age and then decreased. On the other hand, the gene expression of alpha1b-AR subtype did not change over the different stages in relation to that of a housekeeping gene, glyceraldehyde 3-phosphate dehydrogenase, and to total AR. Although rat submandibular glands contain the five AR subtype mRNAs, distinct subtype-specific expression is evident.

Age Factors↗

Genotyping system of GBV-C/HGV type 1 to type 4 by the polymerase chain reaction using type-specific primers and geographical distribution of viral genotypes.

Based on variation in nucleotide sequence of 5'-untranslated region, GB virus type C (GBV-C) and hepatitis G virus (HGV) can be classified into three major genotypes. In addition to this classification, a novel genotype of GBV-C/HGV was identified and designated type 4. However, genotyping of GBV-C/HGV has been hampered by the lack of suitable assays. In this study, a simple and precise genotyping system based on PCR using the type-specific primers was developed for the determination of genotypes 1, 2, 3, and 4 of GBV-C/HGV. A total of 235 serum samples obtained from 11 different countries were tested by our PCR genotyping system of GBV-C/HGV. The results revealed that type 1 was prevalent mainly in Ghana, type 2 was prevalent in the USA, Spain, Egypt, Nepal and Myanmar, type 3 was prevalent in Japan and Bolivia, and type 4 was prevalent in Vietnam and Myanmar among the countries investigated in the present study. To confirm the specificity of the results of genotyping by PCR, phylogenetic analysis in the 5'-untranslated region of GBV-C/HGV was undertaken in 99 serum samples. By this analysis, the specificity of the genotyping system by PCR was confirmed. This assay system may be useful for rapid typing of GBV-C/HGV RNA when either epidemiological or transmission studies of this agent are carried out.

DNA Primers↗

Sustained phosphorylation of mitogen-activated protein kinase is required for basic fibroblast growth factor-mediated axonal branch formation in cultured rat hippocampal neurons.

Basic fibroblast growth factor (bFGF) has been reported to promote the formation of axonal branches in cultured brain neurons. In the present study, we investigated whether the mitogen-activated protein kinase (MAPK) cascade was involved in this action of bFGF in cultured rat hippocampal neurons. Recombinant human bFGF (0.1-10 ng/ml) induced phosphorylation of p44/42 MAPK in a concentration and time-dependent manner. The phosphorylation of p44/42 MAPK occurred rapidly within 5 min after addition of bFGF, and lasted for 48 h. The bFGF-induced phosphorylation of p44/42 MAPK and axonal branch formation were both blocked by simultaneous addition of U0126 and PD98059, specific inhibitors of MAPK kinases. Furthermore, when U0126 and PD98059 were added 24 h after bFGF, phosphorylation of p44/42 m MAPK was decreased, and axonal branch formation was stopped. These results suggest that sustained activation of the MAPK cascade is required for bFGF-mediated axonal branch formation.

Animals↗

Primary culture of rat taste bud cells that retain molecular markers for taste buds and permit functional expression of foreign genes.

Taste buds are constituted of several kinds of cells which have distinct characteristics and play different roles. In this study, we have established an in vitro culture system by optimizing the method for isolating the cells and by selecting culture media and reagents effective for cell viability and adhesion. As a result, the taste bud cells were adhesive and viable for over 3 days when cultured onto Matrigel-coated dishes in medium based on keratinocyte growth medium. The cells retained molecular markers for both the cytoskeleton and intracellular signaling such as cytokeratin 8 and phospholipase Cbeta2. In addition, three intracellular signaling molecules, gustducin, phospholipase Cbeta2, and inositol 1,4,5-trisphosphate receptor type 3, are expressed in the same correlation as those in vivo, although the ratio of signaling molecule-positive cells vs. total cells was somewhat lower in the culture than in vivo. Next, we tried several methods to introduce foreign genes into the cells, and obtained a greater than 90% efficiency of introduction using an adenovirus vector. Finally, we show that an exogenously expressed myc-tagged alpha1A-adrenoceptor sorts into the plasma membrane, and transduces a ligand-dependent signal resulting in intracellular [Ca(2+)] increase in about half of the infected cells. These results suggest that taste bud cells after 3 days of culture retain characteristic molecular markers, and may prove useful for describing the molecular and physiological features of taste bud cells, and that these cells can be further manipulated by adenovirus-mediated gene introduction.

Animals↗

An experimental study on the course of trans-synaptic propagation of neural activity and plasticity in the hippocampus in kainate-induced epilepsy.

To investigate the course of trans-synaptic propagation of neural activity and plasticity in temporal lobe epilepsy, time-dependent changes in the level of synapsin I, a synaptic vesicle protein that is a marker of enhanced synaptic activity and synaptogenesis, were examined following kainate-induced epileptic status in rats. Compared with the control, the level of synapsin I protein increased in the bilateral stratum oriens of CA3 (28.8-40.2%) and CA1 (28.0-34.6%), and the stratum radiatum of CA1 (34.0%) ipsilateral to the injection site at 8 h after intra-amygdala administration of kainate. At 24 h, and 2 and 4 weeks after the kainate treatment, however, synapsin I levels returned to normal levels in most of the regions studied in spite of the extended neural loss in the hippocampus, which suggested the axonal sprouting on the remaining cells. The synapsin I mRNA levels time-dependently decreased bilaterally in CA1-CA3 and the hilus, while no significant changes were observed in the dentate gyrus. These results suggest that the synaptic input to CA3 and CA1 through the stratum oriens was enhanced in this model. A different mode of hippocampal neural activity and plasticity between kainate and kindling models of epilepsy may be stressed.

Animals↗

The effect of positive end-expiratory pressure and continuous positive airway pressure on the oxygenation and shunt fraction during one-lung ventilation with propofol anesthesia.

STUDY OBJECTIVE: To evaluate the effect of positive end-expiratory pressure (PEEP) and continuous positive airway pressure (CPAP) on the oxygenation and shunt fraction during one-lung ventilation (OLV). DESIGN: Prospective clinical study. SETTING: Inpatient thoracic surgery and anesthesia clinic at an University hospital. PATIENTS: 15 patients with esophageal cancer who were scheduled for radical surgery. INTERVENTIONS: Arterial oxygenation, shunt fraction, and hemodynamics were evaluated at 20 min after the start of operation, at 20 minutes after the initiation of OLV under zero end-expiratory pressure (ZEEP), 20 minutes after the application of 4 cm PEEP to the dependent lung, at 20 minutes after OLV under ZEEP, 20 minutes after the application of 4 cm CPAP to the nondependent lung, and again under ZEEP, and after the combined application of PEEP and CPAP to the dependent and nondependent lungs. MEASUREMENTS AND MAIN RESULTS: There were no significant changes in mean pulmonary artery pressure, mean arterial blood pressure, heart rate, mixed venous partial pressure of oxygen, or arterial and mixed venous saturation of oxygen (SVO(2)) during this study. Arterial partial pressure of oxygen (pO(2)) increased and shunt fraction values decreased significantly after the application of PEEP (pO(2); 197.8 +/- 32.9 mmHg, Qs/Qt; 22.9 +/- 5.6%), CPAP (pO(2); 212.6 +/- 15.9 mmHg, Qs/Qt; 22.8 +/- 5.9%), and combination of PEEP and CPAP (pO(2); 222.0 +/- 42.8 mmHg, Qs/Qt; 24.1 +/- 6.4%) compared with ZEEP (pO(2); 128.1 +/- 37.5 mmHg, Qs/Qt; 33.2 +/- 6.8% ). But there were no significant differences regarding oxygenation and shunt fraction during PEEP, CPAP, or the combination of PEEP and CPAP. CONCLUSIONS: The application of PEEP to the dependent lung, CPAP to the nondependent lung, and the combination of PEEP and CPAP, are useful for improving oxygenation and decreasing Qs/Qt.

Adult↗

Development of a transportable incubator for autoradiographic experiments with positron emitter-labeled tracers in living brain tissues.

For autoradiography using positron emitter-labeled tracers in living human brain tissues, samples have to be transported to a positron emission tomography (PET) facility. We have developed a transportable apparatus in which slices are incubated in Krebs-Ringer medium with 95% O(2)/5% CO(2) at 34 degrees C. The incubator comprises a polystyrene foam container, inner chamber, heater, thermostat and battery. The container is sealed and oxygen gas is supplied through a filter (pore size 0.2 microm). The gas is evacuated from the container through the filter, to avoid bacterial contamination. Slices (330 microm) of rat brain were arranged on the nylon net of the inner chamber, and lightly fixed in place by covering them with a fine nylon net stretched over a stainless steel ring. The incubation was carried out at a temperature of 34 degrees C maintained by a heater and thermostat, and the medium was bubbled with 95% O(2)/5% CO(2) from a portable gas cylinder. The temperature of the medium in the container was well controlled (34.0+/-0.5 degrees C) for up to 200 min with no positional differences. In the dynamic autoradiographic experiment, the rate of uptake of [(18)F]2-fluoro-2-deoxy-D-glucose (FDG) in sections preincubated for 45 min in the transportable apparatus was no different from that in the sections kept in the usual incubating apparatus. This apparatus may enable autoradiography using positron emitter-labeled tracers in living human brain tissues.

Animals↗

Japanese experience of telemedicine in oncology.

We started telemedicine projects from 1990 with a telepathology system within Tsukiji Campus of National Cancer Center. In 1994, we connected Tsukiji Campus and Kashiwa Campus by 6 Mbps optical fiber leased line using IP protocol for data transmission, for teleconference, telepathology, and teleradiology projects. We also started connection of regional cancer centers and are now forming a cancer center network of 14 cancer centers. We are at present organizing 130 teleconferences per year with an attendance of more than 16000 people as summary. We have also used a high-resolution image transferring system, such as SHD (2000 pixelsx2000 pixels resolution) system on one side, and an economical telemedicine system using JAVA and a WWW browser (NCC_image) on the other side. We think that providing information is another field of telemedicine. We began the experimental gopher and WWW service in 1993. We are now providing official up-to-date cancer information for patients and healthcare professionals. We are getting more than 400000 hits per month. We are also providing a teleconference video session which is held every week on the Internet using a Real Video system with synchronized slide presentation on the WWW browser. We are also organizing a Cancer Image Reference Database System including DICOM images with viewer software. This paper is a summary of the telemedicine projects performed at the National Cancer Center.

Community Networks↗

Early decrease of survival signal-related proteins in spinal motor neurons of presymptomatic transgenic mice with a mutant SOD1 gene.

The mechanisms of motor neuronal death in amyotrophic lateral sclerosis (ALS) remain to be unclear. Phosphatidy-linositol 3-kinase (PI3-K) and its main downstream effector, Akt/protein kinase B (PKB) have been shown to play a central role in neuronal survival against apoptosis supported by neurotrophic factors. In order to investigate a possible impairment of survival signaling, we examined expressions of PI3-K and Akt in the spinal cord of the transgenic mice overexpressing a mutant Cu/Zn superoxide dismutase (SOD1) gene, a valuable model for human ALS. Immunoblotting and immunohistochemical analyses showed that the majority of spinal motor neurons lost the immunoreactivities for both PI3-K and Akt in the early and presymptomatic stage that preceded significant loss of the neurons. The present results suggest that an early decrease of survival signal proteins in the spinal motor neurons may account for the subsequent motor neuronal loss in this animal model of ALS.

Amyotrophic Lateral Sclerosis↗

Epithelial secretion of C3 promotes colonization of the upper urinary tract by Escherichia coli.

To assess the role of complement in renal infection, we studied a model of Escherichia coli-induced pyelonephritis in mice deficient in complement components C3 and C4. Renal infection occurred less frequently in C3- and C4-deficient mice compared with wild-type mice. In vitro, renal epithelial cells internalized fewer bacteria in the absence of C3 or in the presence of blockade of C3 bound to the bacteria. Moreover, upregulation of epithelial C3 production by stimulation with lipopolysaccharide enhanced bacterial internalization. Here we provide evidence that uropathogenic E. coli might use host C3 to invade the renal epithelium and that local complement production is sufficient for the bacteria to achieve this effect.

Animals↗