Biomedical subjects

K Yamamura

Publications and source records attributed to K Yamamura.

At least 55 records · Page 3Linked to original sources

PubMed2000

Expression of the inducible form of the nitric oxide synthase gene in the livers of mice with chronic hepatitis.

The interferon-gamma (IFN-gamma) transgenic mouse expresses the exogenous IFN-gamma gene in the liver and develops chronic hepatitis. For the present experiment, four IFN-gamma transgene (+) mice of 48 weeks of age and 16 IFN-gamma transgene (+) mice of 8 weeks of age were used. The four IFN-gamma transgene (+) mice of 48 weeks of age showed significantly elevated plasma alanine aminotransferase (ALT) and expressed the inducible form of nitric oxide synthase (iNOS) mRNA in the liver. Of the 16 IFN-gamma transgene (+) mice of 8 weeks of age, iNOS mRNA was expressed in the livers of three. These three mice exhibited higher plasma ALT levels than the other mice of 8 weeks of age. The present results suggest that iNOS mRNA expression in the liver might be correlated with the progression of hepatitis.

Alanine Transaminase↗

PubMed2000

[Receptor and disease: genetic perspective].

Cells in higher animals communicate with each other using hundreds of kinds signaling molecules. Most of these signaling molecules are secreted from the signaling cell by exocytosis or diffusion. Regardless of the nature of the signal, the target cell responds by a specific protein called a receptor. It specifically binds the signaling molecule and then initiates a response in the target cell. Many genes for receptor molecules have been identified and mutations in many receptors are shown to be related to human diseases. In general, both genetic and environmental factors are involved in determining phenotypes including disease susceptibility. As the relative significance of genetic and environmental factors varies from disease to disease, we need animal models for human diseases to analyze these factors. It is now possible to manipulate mouse embryos to produce transgenic and knockout mice. These mice are quite useful for analyses of receptor functions and pathologic processes of disease development. As we only have about 6,000 mutant mice, we still need to produce at lease 94,000 other kinds of mice. To achieve this goal, we have to carry out random mutagenesis, instead of homologous recombination in ES cells.

Animals↗

PubMed2000

[Lung cancer risk and genetic polymorphism at the glutathione S-transferase P1 locus in male Japanese].

The glutathione S-transferases (GSTs) are widely expressed in mammalian tissues and involved in Phase II detoxification reactions. The GSTs form a supergene family consisting of four distinct families, named alpha (GSTA), mu (GSTM) theta (GSTT) and pi (GSTP). Several of the GST genes are polymorphic in humans and are currently being investigated as possible cancer-risk modifiers. Among the GST genes, we examined GSTP1 polymorphism in exon 5 among male lung cancer patients (n = 86, male Japanese) and male healthy controls (n = 80, male Japanese) by restriction fragment length polymorphism-polymerase chain reaction method. The cancer patients showed frequency of the GSTP1 mutated genotype (individuals having at least one mutant allele, 29.1%) very similar to that of the control subjects (28.8%). After adjusting for smoking status, no association was found between the GSTP1 mutated genotype and lung cancer risk (odds ratio: 0.95; 95% confidence interval: 0.48-1.90). When study subjects were divided into two subgroups based on smoking status, the GSTP1 mutated genotype was not associated with an increased risk of lung cancer among smokers and non-smokers. These results suggest that GSTP1 polymorphism in exon 5 alone may not increase the risk of lung cancer in male Japanese.

Adult↗

PubMed2000

[An analysis of characters and changes of the blood pressure in 29 cases on recurrence of cerebral infarction].

We investigated changes of the blood pressure in 29 stroke patients before stroke recurrence and after recurrence. Additional antihypertensive drugs were not administered to all patients after stroke recurrence. Twenty-five lacunar infarct patients and 4 atherothrombotic infarct patients were included in this study. A significant difference was observed between the systolic blood pressure(SBP) within 4 weeks before recurrence and that just after recurrence(132.8 +/- 17.2 mmHg vs. 157.4 +/- 21.3 mmHg, Wilcoxon rank sum test, p < 0.001), or between the SBP just after and 2 weeks after stroke recurrence(157.4 +/- 21.3 mmHg vs. 138.0 +/- 18.3 mmHg, Wilcoxon rank sum test, p < 0.001). The similar difference was found in the diastolic blood pressure(DBP) or in the mean arterial blood pressure(MABP). There was no significant difference in the SBP, the DBP or the MABP between the patients before and 2 weeks after the stroke recurrence. These results suggested that the elevation of the blood pressure at recurrence decreased spontaneously to the pre-recurrence level of the blood pressure in about 2 weeks.

Aged↗

PubMed1999

Mutant mice lacking Crk-II caused by the gene trap insertional mutagenesis: Crk-II is not essential for embryonic development.

Crk family adapter proteins including Crk-II, Crk-I, and Crk-L consist mostly of SH2 and SH3 domains. Through the interactions between SH2 domain and phosphotyrosine residues and/or between SH3 domain and proline-rich motifs, they are involved in a variety of signaling cascades. Despite their essential roles in the signal transductions, knock-out mice of these molecules have not been reported yet. We performed the gene trap insertional mutagenesis with a trap vector, pU-Hachi, and generated a mutant mice line, Ayu 8104, in which the trap vector was inserted into the c-crk gene. Homozygous Ayu 8104 mice lacked Crk-II and Crk-I transcripts but expressed the truncated Crk proteins retaining one SH2 and one SH3 domain. Since the structure of the truncated proteins was similar to that of Crk-I, the insertion was considered to cause Crk-II-specific disruption. Homozygous mutant mice, however, did not exhibit any obvious abnormalities, suggesting that Crk-family adapters, Crk-II, Crk-I, and Crk-L would redundantly function in the signaling cascades and Crk-II was not apparently essential for embryonic development.

Animals↗

PubMed1999

The occurrence of polymorphism of mannose 6-phosphate/insulin-like growth factor 2 receptor gene in laboratory and wild rats.

Carcinogen-resistant inbred DRH rat strain was established from closed colony Donryu rats in the presence of 3'-methyl-4-dimethylaminoazobenzene (3'-Me-DAB). Despite using 3'-Me-DAB during the stage of selection, the DRH rats developed normally and did not show any spontaneous tumor at over 1 year of age. In the present study, we examined the polymorphism in mannose 6-phosphate/insulin-like growth factor 2 receptor (M6p/Igf2r) gene and found that the DRH rat showed CCC (Proline)-type polymorphism in exon 48 and the Donryu rat had GCC (Alanine) sequence. Since the DRH rat was developed from the Donryu rat, we examined whether this polymorphism in exon 48 of M6p/Igf2r gene was due to mutation that occurred at the stage of selection in the presence of 3'-Me-DAB, using several other laboratory and wild rats. We detected the presence of polymorphism at the same site of the M6p/Igf2r gene among these rats. It is likely that the polymorphism in exon 48 of the M6p/Igf2r gene is present broadly in rats since ancient times and not due to the mutation during the course of selection unless this site is a hot spot for chemical carcinogens.

Alanine↗

PubMed1999

Relationship between the clinical manifestations, computed tomographic findings and the outcome in 80 patients with primary pontine hemorrhage.

The relationship between the clinical manifestations, computed tomographic (CT) findings and the outcome in 80 patients with primary pontine hemorrhage (PPH) was analyzed to clarify factors predicting the outcome. Patients were 58 males and 22 females (60. 5+/-12.9 years old) with PPH. Correlations between the clinical manifestations, CT findings and the outcome was assessed with multivariate regression analysis. The initial level of consciousness and the transaxial size of the hematoma on CT were strongly related to the outcome. In patients with small hematoma (with the transaxial size </=20 mm), pupillary abnormalities of onset and acute hydrocephalus were related to the outcome of PPH.

Aged↗

PubMed1999

A novel transgenic technique that allows specific marking of the neural crest cell lineage in mice.

Neural crest cells are embryonic, multipotent stem cells that give rise to various cell/tissue types and thus serve as a good model system for the study of cell specification and mechanisms of cell differentiation. For analysis of neural crest cell lineage, an efficient method has been devised for manipulating the mouse genome through the Cre-loxP system. We generated transgenic mice harboring a Cre gene driven by a promoter of protein 0 (P0). To detect the Cre-mediated DNA recombination, we crossed P0-Cre transgenic mice with CAG-CAT-Z indicator transgenic mice. The CAG-CAT-Z Tg line carries a lacZ gene downstream of a chicken beta-actin promoter and a "stuffer" fragment flanked by two loxP sequences, so that lacZ is expressed only when the stuffer is removed by the action of Cre recombinase. In three different P0-Cre lines crossed with CAG-CAT-Z Tg, embryos carrying both transgenes showed lacZ expression in tissues derived from neural crest cells, such as spinal dorsal root ganglia, sympathetic nervous system, enteric nervous system, and ventral craniofacial mesenchyme at stages later than 9.0 dpc. These findings give some insights into neural crest cell differentiation in mammals. We believe that P0-Cre transgenic mice will facilitate many interesting experiments, including lineage analysis, purification, and genetic manipulation of the mammalian neural crest cells.

Animals↗

PubMed1999

Mice homozygous for a truncated form of CREB-binding protein exhibit defects in hematopoiesis and vasculo-angiogenesis.

CREB-binding protein (CBP) and the closely related adenovirus E1A-associated 300-kD protein (p300) function as coactivators of transcription factors such as CREB, c-Fos, c-Jun, c-Myb, and several nuclear receptors. To study the roles of CBP in embryonic development, we generated CBP homozygous mutant mouse embryos that expressed a truncated form of CBP protein (1-1084 out of 2441 residues). The embryos died between embryonic days 9.5 (E9.5) and E10.5 and exhibited a defect in neural tube closure. They appeared pale and showed decreases in erythroid cells and colony-forming cells (CFCs) in the yolk sac, suggesting defects in primitive hematopoiesis. Immunohistochemistry with an anti-PECAM antibody showed a lack of vascular network formation. Organ culture of para-aortic splanchnopleural mesoderm (P-Sp) with stromal cells (OP9) showed an autonomous abnormality of putative endothelial precursors, which may induce the microenvironmental defect in hematopoiesis. In addition, these defects were partially rescued by the addition of VEGF to this culture. Our analyses demonstrate that CBP plays an essential role in hematopoiesis and vasculo-angiogenesis.

Animals↗

PubMed1999

Effects of functional disruption of lateral pericentral cerebral cortex on primate swallowing.

Bilateral cold block of the intracortical microstimulation (ICMS)-defined swallow cortex markedly affected the ability of monkeys to carry out swallowing. Significant changes also occurred in swallow-related electromyographic (EMG) activity patterns. These findings provide further evidence that the lateral pericentral cortex plays a critical role in the initiation and regulation of swallowing in the primate.

Animals↗

PubMed1999

[Effect of benzene exposure on hematology and hepatic drug metabolic enzymes in ethanol administrated rats].

There are few reports regarding the effects of benzene and ethanol being administered simultaneously. In our experiments with 4 groups (controls, ethanol, benzene and ethanol plus benzene) Wistar male rats were treated with ethanol (20%) for 5 weeks, and then exposed to benzene (0.26 g/kg) for 5 days per week for 3 weeks. We also investigated the effects of benzene on the body weight, organ weight, peripheral hematology and hepatic drug metabolizing enzymes in the ethanol administrated rats. The liver weight increased significantly, but spleen weight decreased significantly in the benzene exposed group. Hematological examination showed a decrease of leukocyte in the two groups of benzene and ethanol plus benzene in comparison with the controls, but an effect promoted by ethanol was not found. Serum glutamate oxaloacetate transaminase (SGOT) and serum glutamate pyruvate transaminase (SGPT) values were not significantly different in the exposure groups when compared with the controls. The contents of microsomal cytochrome P450 in all the exposed groups showed a tendency to increase, but they were not significantly different in comparison with the controls. On the other hand, hepatic glutathione S-transferase activity in the exposed groups increased significantly.

Alanine Transaminase↗

PubMed1999

Comparison of ES cell fate in sandwiched aggregates and co-cultured aggregates during blastocyst formation by monitored GFP expression.

Markers and the means to detect them are required to monitor the fate of living cells. However, few suitable markers for living cells were known until a green fluorescent protein (GFP) was discovered. We have established mouse embryonic stem (ES) cell lines that express mutant GFP under the chicken beta-actin (CAG) promoter. Using these cell lines, we were able to follow the migration of ES cells during blastocyst formation both in sandwiching and coculture methods, even if only a single ES cell was used. Furthermore, the contribution of ES cells to the inner cell mass (ICM) was easily estimated at the blastocyst stage. We compared sandwiching with coculture aggregation relative to the contribution of the ES cell in the ICM, and the results indicated that there was no difference in the ratios of chimeric embryos having ICM contributed from cultured ES cells. Furthermore, an aggregated single ES cell was able to contribute three or four cells to the ICM at the blastocyst stage. Thus we conclude that one, instead of two, embryos is enough to make aggregation with ES cells, and a single ES cell attached to an embryo is enough to produce germline chimeras. Moreover, we could clearly observe single cell fate during blastocyst formation. This suggests that our established cell line can be used for monitoring single cell fate in vivo. In addition, we have shown that up to five doses of 30 sec of UV irradiation using GFP filters have no effect on the embryonic development.

Animals↗

PubMed1999

Difference in the burst patterns of digastric and mylohyoid activities during feeding in the freely behaving rabbit.

Burst patterns in the digastric, mylohyoid, and masseter muscles and the resultant jaw movement orbits during chewing and swallowing were investigated in the freely behaving rabbit. Activities in the posterior mylohyoid fibers consisted of two continuous bursts. Peaks in the first burst of the posterior fibers occurred in the middle part of opening and preceded the digastric burst. Peaks in the second burst occurred in the final part of opening and coincided with those in the working side of the digastric burst. After removal of the bilateral digastric muscles, the gape size during chewing was largely reduced in the final part of opening and in the early part of closing. The results suggest that (a) the digastric may have a role in opening the mandible widely beyond the rest position but may not have a major role in the control of the horizontal (mediolateral) jaw movement, (b) the posterior mylohyoid fibers may have a function as an elevator of the tongue in the early part of opening, and (c) the posterior mylohyoid fibers may have a function as a depressor of the jaw in the late part of opening. Electromyographic burst in the mylohyoid muscle began with marked activity in the mid-closing phase. The results support a role for the mylohyoid muscle as a leading muscle of swallowing. Swallowing events in the rabbit are easily distinguished from the activities of the mylohyoid muscle and the thyrohyoid muscle.

Animals↗

PubMed1999

Genomic organization and expression analysis of the mouse qkI locus.

qkI, encoding a KH domain-containing RNA binding protein, has been isolated as a candidate gene for the mouse neurological mutation quaking. Here, we describe detailed studies on its genomic structure and expression pattern. We isolated approximately 1 Mb of genomic region containing the quaking locus and determined its genomic organization. The qkI locus contains at least 9 exons spanning approximately 65 kb of DNA. It gives rise to six distinct transcripts encoding, theoretically, five different protein isoforms. Exons 1 through 4 are shared by all the transcripts, whereas coding exons and two distinct 3'-UTRs downstream to the exon 4 are differentially utilized. One isoform has a truncated KH domain and may act as an antagonist to the others. These findings and identification of a single transcription initiation site suggest that differential expression of each transcript is regulated by alternative splicing. Expression of each alternative transcript and protein product was also examined. Two types of transcripts, 5 kb-A and B, are most abundant in the brain of newborn mice and are gradually downregulated thereafter. In contrast, the other three messages, 6 kb, 7 kb-A and B, increase as myelination proceeds and peak at 2 weeks of age, corresponding to the most active stage of myelination. Although the qkI messages and their products are abundant in brain and heart, a lower level of expression was found in various other tissues tested. Alternative transcripts that share the same 3'-UTR showed very similar expression patterns, suggesting a regulatory role of the 3'-UTRs in qkI gene expression.

3' Untranslated Regions↗

PubMed1999

Granulocyte/macrophage colony-stimulating factor and interleukin-3 correct osteopetrosis in mice with osteopetrosis mutation.

Although young mice homozygous for the osteopetrosis (op) mutation usually developed prominent osteopetrosis, its severity was markedly reduced in aged op/op mice. This age-associated reversal of osteopetrosis was accompanied by the expansion of bone marrow cavities and increased numbers of tartrate-resistant acid phosphatase (TRAP)-positive cells and of macrophages in the bone marrow. The TRAP-positive cells were mononuclear and developed ruffled borders and numerous vesicles, vacuoles, and granules. Enzyme-linked immunosorbent assay demonstrated a significant elevation of serum granulocyte/ macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-3 levels in the aged op/op mice. To examine whether GM-CSF and/or IL-3 could correct osteopetrosis in young op/op mice, 5 ng of recombinant murine (rm)GM-CSF and/or 100 ng of rmIL-3 were injected daily into young op/op mice. In these treated young op/op mice, the bone marrow cavities were expanded significantly at 2 weeks after administration, associated with significantly increased numbers of TRAP-positive cells and bone marrow macrophages. TRAP-positive cells increased in number with days after injection. These results suggest that GM-CSF and IL-3 induce the development of osteoclasts to correct osteopetrosis in the op/op mice with aging.

Aging↗

PubMed1999

Changes in reflex responses of the masseter and digastric muscles during sleep in freely behaving rabbits.

The aim of this study was to clarify the modulation of motoneuron excitability in masticatory muscles during sleep. For this purpose, changes in the reflex responses of the masseter and digastric muscles between sleep and wakefulness were studied in freely behaving rabbits. Stimulation of the jaw closing muscle spindle afferents induced the masseteric monosynaptic reflex (MMR). During quiet sleep (QS), which gradually replaced quiet wakefulness, the mean MMR amplitude showed no change. During active sleep (AS). MMR amplitudes were markedly reduced, but they were confounded by the occasional occurrence of facilitation in the amplitude. The facilitatory reflexes were often related to rapid eye movements (REMs). However, the excitatory input probably did not originate from the same region as the REM generator, since the REM and the large MMR did not always occur simultaneously. On the other hand, jaw opening reflexes remained inhibited. The results indicated that there is mainly a difference in the excitability between the two groups of motoneurons during AS; masseter motoneuron activity was inhibited but occasionally facilitated by excitatory inputs occurring in association with REMs, however, the digastric motoneuron activity was remained inhibited. The excitatory inputs may induce dysfunctional muscle contraction of the jaw closing muscles as seen in bruxism.

Animals↗

PubMed1999

Characterization of ocular pharmacokinetics of beta-blockers using a diffusion model after instillation.

PURPOSE: To characterize the ocular pharmacokinetics of beta-blockers (timolol and tilisolol) after instillation in the albino rabbit using a mathematical model that includes a diffusion process. METHODS: The disposition of fluorescein isothiocyanate-dextran (FITC-dextran, molecular weight 4400), timolol, and tilisolol was determined in tear fluid and aqueous humor after instillation or ocular injection in rabbits. The in vivo penetration parameters were estimated by fitting the concentration-time profiles to the Laplace equations based on a diffusion model using MULTI(FILT) program. The in vitro permeability of drugs was measured across cornea using a two-chamber diffusion cell. RESULTS: Concentration-time profiles of drugs in the tear fluid after instillation showed a monoexponential curve. Although a monoexponential curve was observed in the aqueous humor concentration of FITC-dextran after injection into the aqueous chamber, timolol and tilisolol showed a biexponential curve. On the basis of these results, an in vivo pharmacokinetic model was developed for estimation of penetration parameters. The in vitro partition parameters were higher than those of the in vivo parameters. CONCLUSIONS: The ocular absorption of timolol and tilisolol was characterized using an in vivo pharmacokinetic model and in vivo penetration parameters.

Absorption↗

PubMed1999

Increased insulin sensitivity and hypoglycaemia in mice lacking the p85 alpha subunit of phosphoinositide 3-kinase.

The hallmark of type 2 diabetes, the most common metabolic disorder, is a defect in insulin-stimulated glucose transport in peripheral tissues. Although a role for phosphoinositide-3-kinase (PI3K) activity in insulin-stimulated glucose transport and glucose transporter isoform 4 (Glut4) translocation has been suggested in vitro, its role in vivo and the molecular link between activation of PI3K and translocation has not yet been elucidated. To determine the role of PI3K in glucose homeostasis, we generated mice with a targeted disruption of the gene encoding the p85alpha regulatory subunit of PI3K (Pik3r1; refs 3-5). Pik3r1-/- mice showed increased insulin sensitivity and hypoglycaemia due to increased glucose transport in skeletal muscle and adipocytes. Insulin-stimulated PI3K activity associated with insulin receptor substrates (IRSs) was mediated via full-length p85 alpha in wild-type mice, but via the p50 alpha alternative splicing isoform of the same gene in Pik3r1-/- mice. This isoform switch was associated with an increase in insulin-induced generation of phosphatidylinositol(3,4,5)triphosphate (PtdIns(3,4,5)P3) in Pik3r1-/- adipocytes and facilitation of Glut4 translocation from the low-density microsome (LDM) fraction to the plasma membrane (PM). This mechanism seems to be responsible for the phenotype of Pik3r1-/- mice, namely increased glucose transport and hypoglycaemia. Our work provides the first direct evidence that PI3K and its regulatory subunit have a role in glucose homeostasis in vivo.

Animals↗