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Biomedical subjects

T Ichimura

Publications and source records attributed to T Ichimura.

At least 73 records · Page 4Linked to original sources

[Plasma interleukin-10 levels in Kawasaki disease].

Time course of interleukin-10 (IL-10) in plasma of patients with Kawasaki disease was measured. The results are as follow: (1) IL-10 concentration in plasma of patients in acute phase of Kawasaki disease (median 37.8 pg/ml) was high, compared with that in convalescent phase (5.77 pg/ml) or that of control children (10.2 pg/ml). (2) From the examination of IL-10 concentration in plasma collected by following its time course, it was revealed that most of cases with this disease showed transiently high figure in the acute phase, then the figure diminished promptly afterwards.

Acute-Phase Reaction↗

[Circulating interleukin-4 levels in acute phase Kawasaki disease].

We have measured the plasma levels of IL-4 in patients with Kawasaki disease, serially. The results were as follows: The plasma levels of IL-4 from patients with Kawasaki disease in acute phase (median 17 pg/ml) were significantly higher than those from the patients in convalescent phase (median 1.96 pg/ ml, p = 0.001) or from control children (median 7.64 pg/ml, p = 0.001). These findings imply that Th2 (or Tho) cells may be vigorously stimulated at an acute phase of this disease, and that IL-4 produced by these cells may possibly modulate the pathological conditions of this disease.

Acute Disease↗

Identification of the site of interaction of the 14-3-3 protein with phosphorylated tryptophan hydroxylase.

The 14-3-3 protein family plays a role in a wide variety of cell signaling processes including monoamine synthesis, exocytosis, and cell cycle regulation, but the structural requirements for the activity of this protein family are not known. We have previously shown that the 14-3-3 protein binds with and activates phosphorylated tryptophan hydroxylase (TPH, the rate-limiting enzyme in the biosynthesis of neurotransmitter serotonin) and proposed that this activity might be mediated through the COOH-terminal acidic region of the 14-3-3 molecules. In this report we demonstrate, using a series of truncation mutants of the 14-3-3 eta isoform expressed in Escherichia coli, that the COOH-terminal region, especially restricted in amino acids 171-213, binds indeed with the phosphorylated TPH. This restricted region, which we termed 14-3-3 box I, is one of the structural regions whose sequence is highly conserved beyond species, allowing that the plant 14-3-3 isoform (GF14) could also activate rat brain TPH. The 14-3-3 box I is the first functional region whose activity has directly been defined in the 14-3-3 sequence and may represent a common structural element whereby 14-3-3 interacts with other target proteins such as Raf-1 kinase. The result is consistent with the recently published crystal structure of this protein family, which suggests the importance of the negatively charged groove-like structure in the ligand binding.

14-3-3 Proteins↗

Separation of membrane proteins solubilized with a nondenaturing detergent and a high salt concentration by hydroxyapatite high-performance liquid chromatography.

Ceramic hydroxyapatite high-performance liquid chromatography with a solvent containing high concentrations of salts was used for the separation of membrane proteins solubilized under nondenaturing and high salt conditions. The chromatographic conditions were optimized using sodium cholate as the detergent. By this method, most membrane proteins, prepared from rat liver rough microsomes, were effectively resolved from each other with a protein recovery of more than 90%. The method also allowed the single-step purification of the ribosome-binding protein, p34, from a microsomal membrane protein fraction. The good resolution with this method should be applicable to the isolation and characterization of a variety of membrane proteins on the analytical and semipreparative scales. With only the substitution of sodium cholate with other nondenaturing detergents, this method may also be applicable to the purification of membrane proteins requiring such nondenaturing detergents with retention of their biological activities.

Cholic Acid↗

Extraction of fuzzy rules using neural networks with structure level adaptation--verification to the diagnosis of hepatobiliary disorders.

This paper presents the reasoning and learning method for fuzzy rules using structure level adaptation of neural networks. In a usual neural network mechanism, we can observe some behaviors during the learning process. Based on such behaviors of neuron activity, we can generate or annihilate the specified neurons respectively in hidden layer to achieve an overall good system. In the method that we have proposed, we have used a procedure to derive the neuron generation/annihilation automatically, and applied such a procedure to the learning system where the experimental data related to hepatobiliary disorders were used. After learning by using randomly chosen data, the proposed system correctly diagnosed over 70% of cases. According to these results, we can find that fuzzy rules have some relationship with the degree of the input weight vector. As a result, we can assume that fuzzy rules for hepatobiliary disorders are extracted from this learned network.

Algorithms↗

FGF-1 in normal and regenerating kidney: expression in mononuclear, interstitial, and regenerating epithelial cells.

The proximal tubule epithelium regenerates following nephrotoxic damage. To determine the role of fibroblast growth factors (FGFs) in the regeneration of rat proximal tubule epithelial (RPTE) cells, we investigated proliferation, differentiation, and FGF-1 expression in vivo in rat kidney before and after nephrotoxic damage to the proximal tubule epithelium caused by S-(1,1,2,2-tetrafluoroethyl)-L-cysteine administration. In undamaged kidneys, FGF-1 was expressed in distal tubule elements, including cortical and medullary collecting ducts, as well as in blood vessels and glomeruli, but was absent in RPTE. One day after damage, there was an increase in proliferation of surviving proximal tubule epithelial cells and a coincident increase in FGF-1 expression in invading mononuclear cells. After this initial burst of proliferation, FGF-1 expression increased in poorly differentiated vimentin-positive regenerative epithelial cells, indicating that autocrine FGF-1 expression in the regenerative epithelium is a later event in the regeneration process. FGF-1 staining persisted in foci of macrophages, interstitial cells, and nephropathic tubules within areas of interstitial expansion 2 wk after damage. We concluded that transient paracrine and autocrine expression of FGF-1 could play mitogenic and/or morphogenic roles during tubular regeneration. Persistent expression in macrophages, fibroblasts, and nephropathic tubules may be associated with tubular degeneration. FGF-1 expression may be an important contributor to both tubular regeneration and degenerative disease following toxicant exposure.

Animals↗

Extraction of fuzzy rules using neural networks with structure level adaptation and its application to diagnosis for hepatobiliary disorders.

First, this paper presents the reasoning and the learning method for fuzzy rules using structure level adaptation of neural networks. In a usual neural network's mechanism, during learning process of rules, we can observe the following two behaviors: Case 1: If a neural network does not have enough neurons to be satisfied to infer, then the input weight vector will have a tendency to fluctuate greatly, even after a certain long period the learning process. In this case, the network needs to generate a new neuron as its parent's attribute is inherited. Case 2: If a neural network has enough neurons to infer, and even if the input weight vector of each neuron will converge to a certain value, then we shall be able to turn out unnecessary neurons from the network in the calculation. In this case, because it is necessary to delete a redundant neuron to the calculation, the neuron is annihilated without affecting the performance of the network. By observing such behaviors, we can generate or annihilate the specified neuron respectively to achieve an overall good system. In the proposed method, we described a procedure to derive the neuron generation/annihilation automatically and applied the procedure to learning system. Next, we apply such procedure to the learning system in which the experimental data related to hepatobiliary disorders is used. We use a real medical database containing the results of ten biochemical terms test for four hepatobiliary disorders. We have 536 case data, including some errors. After the learning, by using 179 data chosen randomly from database, the proposed system converged to a certain small value and this constructed network has the optimal structure for these teaching data. In addition, we get that the fuzzy rules have some meanings related to the degree of the input weight vector, and the fuzzy rules for hepatobiliary disorders are extracted from the learned network with respect to the degree of input weight vector. Moreover, to verify the validity of the diagnosis of the proposed method, the feed-forward calculation was implemented using extracted fuzzy rules for all databases. As a result, the proposed system correctly diagnosed more than 70%.

Biliary Tract Diseases↗

A nuclear factor containing the leucine-rich repeats expressed in murine cerebellar neurons.

A nuclear protein, termed leucine-rich acidic nuclear protein (LANP), has been isolated from among rat cerebellar proteins whose expression was transiently increased during an early stage of postnatal development. The amino acid sequence, deduced from its cDNA, showed that LANP contains 247 amino acids consisting of two distinct structural domains: the N-terminal domain characterized by "leucine-rich repeat," which is found in many eukaryotic proteins and which potentially functions in mediating protein-protein interactions, and the C-terminal domain characterized by a cluster of acidic amino acids with a putative nuclear localization signal. Immunohistochemical study using an antibody against LANP revealed that the protein is localized mainly in nuclei of Purkinje cells. In the rat cerebellum on postnatal day 7, LANP mRNA was expressed moderately in the external granule and Purkinje cells and weakly in the internal granule cells. The expression in these cells, especially in Purkinje cells, increased in the second postnatal week and thereafter decreased to an adult level. The structural characteristics, localization, and the stage- and cell type-specific expression suggest a potential role of LANP in a signal transduction pathway that directs differentiation of cerebellar neurons.

Aging↗

Differential effects of methylmercury on the phosphorylation of protein species in the brain of acutely intoxicated rats.

The in vivo effect of methylmercury (MeHg) on the phosphorylation in vitro of the brain cytosol fraction was examined in acutely poisoned rats (10 mg/kg/day, for 7 days). The total phosphorylation activity, determined in the presence or absence of protein kinase effectors (Ca2+ and cAMP) and substrates (casein, histone and protein kinase C substrate), did not markedly change with the progress of intoxication. Two-dimensional electrophoretic analysis of the phosphorylated cytosol fractions from control and MeHg-treated rats revealed that (1) the extents of phosphorylation of the 24 major protein species in the control rats differed greatly from each other, (2) the effect of MeHg on the phosphorylation was not uniform regarding the individual 24 proteins or the period of intoxication, and (3) in the symptomatic period, many protein species including tubulin subunits showed elevated phosphorylation, while a few protein species showed decreased phosphorylation. These results suggest that the neurotoxic action of MeHg could be mediated through, at least in part, the modification of functional protein species due to excess phosphorylation that leads to impairment of the normal cellular processes.

Amino Acid Sequence↗

A single Arabidopsis GF14 isoform possesses biochemical characteristics of diverse 14-3-3 homologues.

Arabidopsis cDNA clones of GF14 proteins originally were isolated on the basis of their association with the G-box DNA/protein complex by a monoclonal antibody screening approach. GF14 proteins are homologous to the 14-3-3 family of mammalian proteins. Here we demonstrate that recombinant GF14 omega, one member of the Arabidopsis GF14 protein family, is a dimeric protein that possesses many of the attributes of diverse mammalian 14-3-3 homologues. GF14 omega activates rat brain tryptophan hydroxylase and protein kinase C in a manner similar to the bovine 14-3-3 protein. It also activates exoenzyme S of Pseudomonas aeruginosa as does bovine brain factor activating exoenzyme S (FAS), which is itself a member of 14-3-3 proteins. In addition, GF14 omega binds calcium, as does the human 14-3-3 homologue reported to be a phospholipase A2. These results indicate that a single isoform of this plant protein family can have multiple functions and that individual GF14 isoforms may have multiple roles in mediating signal transductions in plants. However, GF14 omega does not regulate growth in an in vivo test for functional similarity to the yeast 14-3-3 homologue, BMH1. Thus, while a single plant GF14 isoform can exhibit many of the biochemical attributes of diverse mammalian 14-3-3 homologues, open questions remain regarding the physiological functions of GF14/14-3-3 proteins.

14-3-3 Proteins↗

Three-dimensional architecture of the tubular endocytic apparatus and paramembranous networks of the endoplasmic reticulum in the rat visceral yolk-sac endoderm.

The three-dimensional architecture of the tubular endocytic apparatus and the endoplasmic reticulum in the rat yolk-sac endoderm was investigated after loading with horseradish peroxidase-conjugated concanavalin A by intrauterine administration. After 30 min, small vesicles (50-150 nm in diameter), small tubules (80-100 nm in diameter) and large vacuoles (0.2-1.0 microns in diameter) in the apical cytoplasm were labeled with the tracer, but lysosomes (1.0-3.5 microns in diameter) in the supranuclear cytoplasm were not labeled until 60 min after loading. Stereo-viewing of the labeled small tubules in thick sections revealed that they were not isolated structures but formed three-dimensional anastomosing networks, which were also confirmed by scanning electron microscopy after maceration with diluted osmium tetroxide. Their earlier labeling with the endocytic tracer, localization in the apical cytoplasm and three-dimensional network formation indicated that the labeled small tubules represented tubular endosomes (tubular endocytic apparatus). These well-developed membranous networks provided by the tubular endosomes are suggested to facilitate the receptor-mediated endocytosis and transcytosis of the maternal immunoglobulin in the rat yolk-sac endoderm. Scanning electron microscopy further revealed lace-like networks of the smooth endoplasmic reticulum near the lateral plasma membrane. Their possible involvement in transport of small molecules or electrolytes is discussed.

Animals↗

Thyroid function in children with nephrotic syndrome.

The thyroid function of seven children with untreated nephrotic syndrome who had a normal serum creatinine concentration was compared with that of the same patients in remission and age-matched controls. There was a significant decrease in serum thyroxine (T4), tri-iodothyronine (T3) and thyroid-binding globulin (TBG) concentrations in untreated nephrotic children compared with the same patients in remission and age-matched controls. Most values for serum free T4, free T3 and thyroid-stimulating hormone (TSH) in the patients with nephrosis were within the normal range. However, the mean serum free T4 and free T3 concentrations were significantly (P < 0.05) lower in the untreated patients than in the same patients in remission, and the mean serum TSH concentrations were significantly (P < 0.05) higher in the untreated patients than in the same patients in remission. There were massive urinary losses of T4, T3, TBG, free T4 and free T3 in the untreated nephrotic children compared with the same patients in remission and age-matched controls. The daily urinary protein excretion showed a positive correlation with the urinary T4, T3, free T4, free T3 and TBG excretion. Furthermore, the urinary protein excretion showed a negative correlation with the serum T4, T3, free T4, free T3 and TBG levels. There was a negative correlation between serum albumin and serum TSH. These findings provide evidence of mild hypothyroidism in children with untreated nephrotic syndrome, partly because of losses of T4, T3, free T4, free T3 and TBG into the urine.

Child↗

Molecular cloning of rat cDNAs for the zeta and theta subtypes of 14-3-3 protein and differential distributions of their mRNAs in the brain.

We isolated from the rat brain two cDNA clones encoding the zeta and theta subtypes of the 14-3-3 protein. Both clones encoded 245 amino acid sequences, which share a high sequence homology with each other and also with other subtypes of the 14-3-3 protein. The distribution of their mRNAs was determined in the developing brain, by in situ hybridization with subtype-specific oligonucleotide probes. At embryonic day 18, the zeta and theta subtype mRNAs were expressed at high levels throughout the brain and the spinal cord. Distribution patterns of the two mRNAs were distinct in the brain at postnatal day 21. The zeta subtype mRNA was distributed widely in the brain gray matter, and high levels of the transcripts were detected in various brain regions, including the neocortex, hippocampus, caudate-putamen, thalamus, cerebellar cortex, and several brainstem nuclei. On the other hand, high signal levels of the theta subtype mRNA in the gray matter were restricted to the cerebellar cortex and the hippocampus. In addition, significant signals for the theta subtype mRNA were found over the white matter, where cell bodies of glial cells are populated. The wide gene expression of the zeta and theta subtypes suggests their fundamental and essential role in the brain function, but the degrees of functional involvement by the respective subtypes would be heterogeneous between neuron and glia, and also among neuron types.

14-3-3 Proteins↗

Stable and temperature-sensitive transformation of rat kidney epithelial cells suppresses expression of acidic fibroblast growth factor 1 but activates secretion of fibroblast growth factor 3 (int-2) and vascular endothelial growth factor.

Rat kidney proximal tubule epithelial cells (RPTE) in primary culture express acidic fibroblast growth factor 1 (FGF-1). Transformation of RPTE by SV40 (SV-RPTE) suppressed FGF-1 expression but activated secretion of FGF-like factor(s). SV-RPTE conditioned medium contained growth-promoting activity for SV-RPTE and human umbilical vein endothelial cells, indicating that both autocrine and angiogenic factors were secreted. Reverse transcriptase-polymerase chain reaction and Northern analysis for various FGFs showed that only FGF-3, also known as int-2, mRNA was expressed in SV-RPTE. In addition, expression of mRNA for the heparin-binding angiogenic factor vascular endothelial growth factor (VEGF) increased dramatically in SV-RPTE. Physical characterization of the activity in the SV-RPTE conditioned medium suggested that FGF-3 and VEGF contributed the autocrine and angiogenic activities, respectively. We also investigated FGF-3 and VEGF secretion in temperature-sensitive (ts) SV40-transformed RPTE. tsSV-RPTE had transformed properties resembling those of SV-RPTE only at the permissive temperature (33 degrees C), e.g., increased growth potential and anchorage-independent growth. FGF-1 was expressed only at the nonpermissive temperature. VEGF mRNA levels and secretion of the human umbilical vein endothelial cell growth-promoting activity were reduced by switching tsSV-RPTE cells from 33 degrees to 39 degrees C. However, FGF-3 mRNA levels were not affected significantly by the temperature switch suggesting that activation of VEGF and FGF-3 occurs through different mechanisms. These results indicate that FGF-1 expression in RPTE is suppressed by SV40 transformation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

[Chromogranin A like immunoreactivity in respiratory tissues of ovalbumin-sensitized guinea pigs].

Chromogranin A like immunoreactivity (CGA-IR) was measured by a specific radioimmunoassay using N-terminal specific antiserum R-0763 in respiratory tissues of guinea pigs. All guinea pig were actively immunized by recurrent Ovalbumin (OA)-inhalation and divided into two groups; one was challenged by OA, and the other inhaled saline as a control. These groups were studied for respiratory resistance and sacrificed for measurement of CGA-IR concentrations in the trachea, major bronchus and lower lung. In the control group, CGA-IR level was 0.4-2 pmol/g wet weight of tissue, and its distribution order was lower lung > trachea > major bronchus. In the OA-challenged group, provocation clearly induced significant elevation of CGA-IR in the trachea and main bronchus coinciding with elevation of respiratory resistance. In the lower lung, on the other hand, the increase in CGA-IR was not significant. These results suggest that experimental immediate asthmatic response (IAR) possibly acts as a stressor to the sympathetic nervous systems in guinea pig air-ways.

Airway Resistance↗

Ribosome-binding protein p34 is a member of the leucine-rich-repeat-protein superfamily.

Protein p34 is a non-glycosylated membrane protein characteristic of rough microsomes and is believed to play a role in the ribosome-membrane association. In the present study we isolated cDNA encoding p34 from a rat liver cDNA library and determined its complete amino acid sequence. p34 mRNA is 3.2 kb long and encodes a polypeptide of 307 amino acids with a molecular mass of about 34.9 kDa. Primary sequence analysis, coupled with biochemical studies on the topology, suggested that p34 is a type II signal-anchor protein; it is composed of a large cytoplasmic domain, a membrane-spanning segment and a 38-amino-acid-long luminally disposed C-terminus. The cytoplasmic domain of p34 has several noteworthy structural features, including a region of 4.5 tandem repeats of 23-24 amino acids. The repeated motif shows structural similarity to the leucine-rich repeat which is found in a variety of proteins widely distributed among eukaryotic cells and which potentially functions in mediating protein-protein interactions. The cytoplasmic domain also contains a characteristic hydrophilic region with abundant charged amino acids. These structural regions may be important for the observed ribosome-binding activity of the p34 protein.

Amino Acid Sequence↗