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Biomedical subjects

M Tamura

Publications and source records attributed to M Tamura.

At least 199 records · Page 11Linked to original sources
PubMed1999

Mutation of Pten/Mmac1 in mice causes neoplasia in multiple organ systems.

Pten/Mmac1+/- heterozygous mice exhibited neoplasms in multiple organs including the endometrium, liver, prostate, gastrointestinal tract, thyroid, and thymus. Loss of the wild-type allele was detected in neoplasms of the thymus and liver. Surprisingly, tumors of the gastrointestinal epithelium developed in association with gut lymphoid tissue. Tumors of the endometrium, thyroid, prostate, and liver were not associated with lymphoid tissue and appeared to be highly mitotic. In addition, these mice have nonneoplastic hyperplasia of lymph nodes that was caused by an inherited defect in apoptosis detected in B cells and macrophages. Examination of peripheral lymphoid tissue including lymphoid aggregates associated with polyps revealed that the normal organization of B and T cells was disrupted in heterozygous animals. Taken together, these data suggest that PTEN is a regulator of apoptosis and proliferation that behaves as a "landscaper" tumor suppressor in the gut and a "gatekeeper" tumor suppressor in other organs.

Animals↗
PubMed1999

Identification of DERMO-1 as a member of helix-loop-helix type transcription factors expressed in osteoblastic cells.

Several members of the basic helix-loop-helix (bHLH) type of transcription factors have now been reported, and almost every member of this class has been implicated in transcriptional regulation in cell type determination and differentiation. Previously, we reported that dominant negative HLH proteins are involved in osteoblastic phenotype expression, such as osteocalcin, and hence differentiation (Tamura and Noda [1994] J. Cell Biol. 126:773-782). In this work, we used degenerate PCR cloning in order to identify cDNA clones encoding bHLH proteins expressed in osteoblastic osteosarcoma ROS17/2.8 cells. Sequence analyses of the 47 clones revealed that 11 clones encoded products with a characteristic motif of the bHLH transcription factor family. Of these clones, sequences in the amplified region of seven clones were homologous to the mouse twist, and three clones were homologous to the mouse twist-related HLH protein, Dermo-1. To confirm Dermo-1 mRNA expression in osteoblastic cells, we performed reverse transcription polymerase chain reaction (RT-PCR) analysis using mRNA from ROS17/2.8 cells and MC3T3-E1 cells by Dermo-1 specific primers and Northern blot analysis. These analyses demonstrated that Dermo-1 mRNA was expressed in these osteoblast-like cell lines. Nucleotide sequence analysis of the partial rat Dermo-1 cDNA cloned from ROS17/2.8 library revealed that it has the highest degree of homology with the mouse Dermo-1 cDNA, and the partial amino acid sequence deduced from the obtained rat Dermo-1 was identical with the corresponding region of the mouse Dermo-1 amino acid sequence. To further examine the role of Dermo-1 in the regulation of osteoblastic differentiation, we examined mRNA levels of Dermo-1 and twist in C3H10T1/2 cells treated with recombinant human bone morphogenetic protein (rhBMP)-2. Using the RT-PCR method, the mRNA levels of Dermo-1 and twist were found to be decreased by the treatment with rhBMP-2 in C3H10T1/2 cells. We also observed that the mRNA level of Dermo-1 was decreased about fourfold by the treatment with rhBMP-2 in C3H10T1/2 cells by Northern blot analysis. Moreover, Dermo-1 mRNA was detected at lower levels in 21-day-old differentiated MC3T3-E1 cells compared with 3-day-old undifferentiated MC3T3-E1 cells. These results suggested that Dermo-1 could be involved in the osteoblastic differentiation in a negative manner.

Amino Acid Sequence↗
PubMed1999

Augmented expression of macrophage migration inhibitory factor (MIF) in the telencephalon of the developing rat brain.

Macrophage migration inhibitory factor (MIF) has been characterized as a proinflammatory cytokine, hormone, and immunomodulator. We previously demonstrated the distribution of MIF in the adult rat brain and revealed its expression in neurons as well as glial cells. In this study, we investigated the localization of MIF in the developing rat brain from embryonic day (ED) 13 to ED19, and after birth from postnatal day (PD) 1 to PD28 using both immunohistochemistry and in situ hybridization. On ED16, the signals of MIF mRNA were high in the ventricular, subventricular, and intermediate zones of the telencephalon, in which 'neuropoietic' progenitor cells proliferate and migrate from the ventricular zone to the superficial layer in the cerebral cortex. The mRNA expression was detected throughout the postnatal period, and intense signals of the transcript were seen in the ventricle and in layers I, II and III of the cerebral cortex at PD5. Similarly, positive staining of MIF protein was seen in the ventricular zone by immunohistochemical analysis, although the positively stained area appeared to be smaller than the mRNA expression. Taken together, these results suggest that the increase in MIF mRNA in the developing brain reflects the growth and maturation of neurons and glial cells.

Animals↗
PubMed1999

Guanosine 5'-O-(3-thiotriphosphate)-induced O-2 generation in permeabilized neutrophils requires protein kinase C and phospholipase C but not tyrosine kinase or phospholipase D.

Guanosine 5'-O-(3-thiotriphosphate) (GTPgammaS) induces respiratory burst (O-2 generation) in permeabilized human neutrophils. The signal pathway from GTPgammaS to the enzyme responsible for O-2 generation (NADPH oxidase) is not well defined. To elucidate the signaling pathway activated by GTPgammaS, we used selective inhibitors to test for the involvement of several enzymes, comparing the effects of these inhibitors on fMet-Leu-Phe (fMLP) activation. GTPgammaS-induced respiratory burst was not influenced by genistein, a selective inhibitor of tyrosine kinase, while fMLP-induced response was completely abolished. The respiratory burst by GTPgammaS was efficiently inhibited by the protein kinase C inhibitor GF109203X even more than fMLP activation. The mitogen-activated protein kinase (MAPK) kinase inhibitor PD098059 showed a partial inhibition of both GTPgammaS and fMLP activation. Wortmannin, an inhibitor of phosphatidylinositol 3-kinase, completely blocked fMLP activation, but had no effect on the GTPgammaS-induced respiratory burst. Using U73122, phospholipase C is shown to be essential in GTPgammaS signaling as well as fMLP signaling. Butanol blocked fMLP signaling but not GTPgammaS signaling, indicating that only fMLP activation involves phospholipase D. These results suggest that there are several differences between GTPgammaS- and fMLP-induced activation, but both activators share a common pathway including phospholipase C, protein kinase C, and MAPK kinase.

Cell Membrane Permeability↗
PubMed1999

Tumor suppressor PTEN inhibition of cell invasion, migration, and growth: differential involvement of focal adhesion kinase and p130Cas.

PTEN/MMAC1 is a major new tumor suppressor gene that encodes a dual-specificity phosphatase with sequence similarity to the cytoskeletal protein tensin. Recently, we reported that PTEN dephosphorylates focal adhesion kinase (FAK) and inhibits cell migration, spreading, and focal adhesion formation. Here, the effects of PTEN on cell invasion, migration, and growth as well as the involvement of FAK and p130 Crk-associated substrate (p130Cas) were investigated in U87MG glioblastoma cells missing PTEN. Cell invasion, migration, and growth were down-regulated by expression of phosphatase-active forms of PTEN but not by PTEN with an inactive phosphatase domain; these effects were correlated with decreased tyrosine phosphorylation levels of FAK and p130Cas. Overexpression of FAK concomitant with PTEN resulted in increased total tyrosine phosphorylation levels of FAK and p130Cas and effectively antagonized the effects of PTEN on cell invasion and migration and partially on cell growth. Overexpression of p130Cas increased total tyrosine phosphorylation levels of p130Cas without affecting those of FAK; however, although p130Cas could reverse PTEN inhibition of cell invasion and migration, it did not rescue cell growth in U87MG cells. In contrast to FAK, p130Cas could not be shown to interact with PTEN in cells, and it was not dephosphorylated directly by PTEN in vitro. These results suggest important roles of PTEN in the phenotype of tumor progression, and that the effects of PTEN on cell invasion, migration, and growth are mediated by distinct downstream pathways that diverge at the level of FAK.

Cell Adhesion Molecules↗
PubMed1999

Expression of major urinary protein genes in the nasal glands associated with general olfaction.

Gene expression of major urinary protein (MUP) mRNAs was examined in the mouse nasal tissue. By polymerase chain reaction, we identified two cDNA segments encoding MUP 4 and MUP 5 genes in the nose. The expression level of both MUP 4 and 5 mRNAs in the nasal tissue was very high and exceeded that of the liver. Liver MUPs are excreted into the urine and are known to play an important role in pheromonal communication. We showed that nose and liver MUPs were composed of different subtypes of MUPs and that nose MUP mRNAs was detected in prepubescent periods when liver MUP mRNAs had not yet been transcripted. In situ hybridization revealed that nose MUP mRNAs are localized in the lateral wall and nasal septum and their expression pattern is identical to that of rat odorant-binding protein (OBP)-I. We also identified cDNA of mouse OBP-II gene from the nasal tissue and showed that the expression pattern of MUP gene was identical to that of OBP-II gene in the nose. These histological data indicate that nose MUPs are favorable for catching odorant molecules rather than pheromones, and may share their function with OBPs.

Amino Acid Sequence↗
PubMed1999

Differential production of MCP-1 and cytokine-induced neutrophil chemoattractant in the ischemic brain after transient focal ischemia in rats.

Chemokines have been shown to play an important role in leukocyte infiltration into ischemic lesions. Recently, the increased expression of monocyte chemoattractant protein-1 (MCP-1) and cytokine-induced neutrophil chemoattractant (CINC) was observed in experimental stroke models where infiltrated leukocytes were supposed to induce tissue injury, however, the protein level and time course of these chemokines have not been fully elucidated. Therefore, we analyzed the time-dependent production of MCP-1 and CINC in the rat brain after transient middle cerebral artery occlusion (MCAO) by means of specific enzyme-linked immunosorbent assay systems. The MCP-1 levels in the ipsilateral hemispheres increased from 6 h, peaked at 2 days, and thereafter gradually decreased. The peak MCP-1 concentration was 89.2+/-28.2 ng/g tissue wet weight (mean +/- SEM, n = 5, 49.3-fold greater than the contralateral value at the same time, P < 0.05), which is supposed to be high enough to exert its biological effects. In contrast, the maximum CINC concentration that corresponded to 2.9+/-0.7 ng/g tissue wet weight (mean +/- SEM, n = 5, 55.0-fold greater than the contralateral value at the same time, P < 0.05), was observed at 6 h. In addition, we confirmed the temporal profile of leukocyte subtypes that infiltrated into the ischemic brain, thus, neutrophil infiltration occurred at early stages (1-3 days), followed by massive infiltration of macrophages at later stages (2-7 days). These studies suggest that MCP-1 in cerebral ischemia actually plays a significant role in the migration of macrophages into the lesion and that the differential temporal production of these chemokines contributes to the regulation of infiltrated leukocyte subtypes.

Animals↗
PubMed1999

Suppressive effect of anticardiolipin antibody on the proliferation of human umbilical vein endothelial cells.

OBJECTIVE: To determine whether sera that show a positive anticardiolipin test result have a suppressive effect on the proliferation of endothelial cells collected from human umbilical veins. DESIGN: Retrospective in vitro study. SETTING: University hospital outpatient clinic for the treatment of infertility. PATIENT(S): Thirteen patients with recurrent fetal miscarriages who were positive for anticardiolipin antibody and 14 patients with recurrent miscarriages who were negative for anticardiolipin antibody. INTERVENTION(S): Serum was obtained from each patient. MAIN OUTCOME MEASURE(S): Suppressive effect of the sera on the culture of human umbilical vein endothelial cells. RESULT(S): The proliferation of human umbilical vein endothelial cells decreased significantly in cultures that contained sera showing positive anticardiolipin antibody activity that had been collected from patients with recurrent fetal miscarriages. CONCLUSION(S): The results strongly suggest that anticardiolipin antibody has a suppressive effect on the proliferation of human umbilical vein endothelial cells.

Abortion, Habitual↗
PubMed1999

Iron depletion prevents adenine nucleotide decomposition and an increase of xanthine oxidase activity in the liver of the Long Evans Cinnamon (LEC) rat, an animal model of Wilson's disease.

The Long Evans Cinnamon (LEC) rat, which accumulates excess Cu in the liver as in patients with Wilson's disease, is a mutant strain displaying spontaneous hepatitis. It was reported that Fe, like Cu, increases in the liver and that the severity of hepatitis is modified by Fe in the diet. In this experiment, oxidative stress increased by Fe was investigated before the onset of hepatitis. To examine the effect of Fe on the progress into hepatitis, LEC female rats were fed an Fe-regular (Fe 214 microg/g; Fe(+) group) or an Fe-restricted (Fe 14 microg/g; Fe(-) group) diet from 53 days of age for 35 days. Fischer rats were also fed as control animals. Adenine nucleotide decomposition was determined as an index of oxidative stress based on xanthine oxidase activity. The size of the hepatic pool of adenine nucleotides (ATP+ADP+AMP) was significantly smaller in LEC rats than Fischer rats. The energy charge (ATP+0.5ADP)/(ATP+ADP+AMP) was smaller in Fe(+) groups than in Fe(-) groups. In the LEC rat liver, the Fe concentration in the Fe(+) group was 160% of that in Fe(-) group and the correlation coefficient between the hepatic Fe concentration and the energy charge was significant. In this strain, an increase of xanthine oxidase activity resulted in an increase of xanthine, an oxidized metabolite of hypoxanthine in the liver. The results suggest the involvement of the Fe in the progression into hepatitis in the LEC rat, even if the dietary Fe concentration is similar to that of commercial diet.

Adenine Nucleotides↗
PubMed1999

Involvement of cholinergic and GABAergic systems in the reversal of memory disruption by NS-105, a cognition enhancer.

The effects of (+)-5-oxo-D-prolinepiperidinamide monohydrate (NS-105) on the scopolamine-, electrolytic lesion of the nucleus basalis magnocellularis (NBM)-, AF64A-, baclofen-, cerebral ischemia- and electroconvulsive shock (ECS)-induced memory disruption in the passive avoidance response or radial arm maze tasks were investigated in rats. The effects of NS-105 were compared with those of aniracetam, bifemelane, idebenone, and indeloxazine in two tasks of the passive avoidance response. Furthermore, effects of NS-105 on in vivo release of acetylcholine (ACh) in the cerebral cortex, high-affinity choline uptake (HACU) of the cerebral cortex in rats with lesion of NBM, HACU of the hippocampus in rats treated with pentobarbital and activity of choline acetyltransferase (ChAT) of the cerebral cortex in rats with lesion of NBM were examined. NS-105 showed antiamnestic actions in a variety of animal models of cholinergic dysfunction employed in this study. Aniracetam improved memory disruption caused by scopolamine, but bifemelane, idebenone, and indeloxazine did not. NS-105 (10 mg/kg) showed the increase of ACh release from the cerebral cortex and the enhancement of HACU both in the cerebral cortex and hippocampus, but showed no change in activity of ChAT. NS-105 also reversed memory disruption induced by baclofen, a potent GABA(B) receptor agonist, but all of reference drugs did not. These results suggest that antiamnestic action of NS-105 is due to the facilitation of cholinergic neuronal activity and the suppression of GABA(B) receptor-mediated responses.

Acetylcholine↗
PubMed1999

Serial echocardiographic and Doppler evaluation of left ventricular systolic performance and diastolic filling in premature infants.

To examine the effects of somatic growth on left ventricular systolic and diastolic functions in premature infants, we performed serial two-dimensional, M-mode and Doppler echocardiograms of 18 premature infants in the early neonatal period (mean of 14 days) and on the day when they were discharged from the hospital (mean of 94 days). The relation between rate-corrected mean velocity of fiber shortening and end-systolic wall stress relation was used as a load-independent estimate of contractility. Diastolic function was assessed by transmitral Doppler filling velocity patterns. The end-systolic wall stress was significantly lower in the first examination than in the second examination (38+/-8 vs. 46+/-10 g/cm2, P < 0.005). However, there was no significant difference in the rate-corrected velocity of fiber shortening between the two examinations. An inverse linear relation between the end-systolic wall stress and the rate-corrected mean velocity of fiber shortening was found in each examination. The slopes of these two regression lines showed no significant difference, suggesting that the left ventricular contractility is on the same level during this study period. The transmitral Doppler flow velocity patterns markedly altered between the two observations. The peak E wave, peak A wave, peak E/A ratio, flow velocity time integral of E/A wave, the first third filling fraction, and the normalized peak filling rate to stroke volume increased significantly during the study period (78+/-14 vs. 31+/-9 cm/s, 61+/-12 vs. 33+/-7 cm/s, 1.29+/-0.18 vs. 0.93+/-0.19, 1.75+/-0.58 vs. 1.17+/-0.24, 0.42+/-0.07 vs. 0.37+/-0.05, and 9.48+/-1.71 vs. 7.30+/-0.96/s, respectively, P < 0.001), suggesting a relative shift of Doppler filling into the early diastole. We demonstrated that the age- and growth-related alterations in the transmitral Doppler filling patterns occurred dramatically without changes in contractility during the first 3 months after birth. This diastolic filling change may be related to the age-related maturation in the left ventricular diastolic properties.

Analysis of Variance↗
PubMed1999

A method for detection of a cytokine and its mRNA in the central nervous system of the developing rat.

We report here an effective and concise method to determine the localization of macrophage migration inhibitory factor (MIF), a proinflammatory cytokine, and its mRNA in the central nervous system of pre- and postnatal rats. This method allows for double staining to demonstrate localization of different molecules on the same tissue specimen at the levels of mRNA and proteins by in situ hybridization and immunohistochemistry, respectively. Additionally, the present method gives results more quickly than the conventional isotopic techniques. By use of this method, we carried out immunohistochemistry with an anti-rat MIF polyclonal antibody and demonstrated positive staining using the avidin-biotin complex method (ABC method). To detect its mRNA, we performed nonradioactive in situ hybridization using a digoxigenin (DIG)-labeled RNA probe prepared from a full length fragment of rat MIF cDNA. MIF was strongly expressed in the telencephalon on embryonic day 16. Non-radioactive in situ hybridization with a DIG-labeled RNA probe as well as the immunohistochemistry described here could be applicable to characterize localization of mRNA and proteins of different molecules on the same tissue specimen.

Animals↗
PubMed1999

Recurrent bladder adenocarcinoma in an ileal conduit stoma: a case report.

BACKGROUND: The first case of a patient with recurrent adenocarcinoma in an ileal conduit stoma 7 months after radical cystectomy is reported. RESULTS/DISCUSSION: The most likely explanation of this case is hematogenous metastasis based on the clinical diagnosis and the pathological immunostaining examination of cytokeratin.

Aged↗
PubMed1999

7-ethoxycoumarin deethylation activity in perfused isolated rat brain.

7-ethoxycoumarin (7-EC) deethylation activity was measured in the perfused rat brain in situ. Infusion of 7-EC into a brain through an internal carotid artery resulted in the formation of 7-hydroxycoumarin (7-HC) and its conjugates in the effluent perfusate collected from the superior vena cava. The rate of formation of products was 200 nmol/h/g when 130 microM 7-EC was infused. This value was much higher (more than 100 times) than that determined from the brain microsomal activity ( approximately 1 nmol/h/g), indicating that the activity determined with microsomes was an underestimate. This value was comparable to the activity in the perfused liver (30-50%), suggesting that drug metabolizing enzymes can play important roles within the brain. Pretreatment of rats with P-450 inducers such as phenobarbital and beta-naphthoflavone increased the deethylation activity in the perfused brain, as in the perfused liver. We conclude that the perfused brain is suitable for evaluating drug metabolizing activities under physiological conditions.

Animals↗