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Biomedical subjects

T Mizuno

Publications and source records attributed to T Mizuno.

At least 343 records · Page 19Linked to original sources
PubMed1994

Molecular cloning and expression of an isomalto-dextranase gene from Arthrobacter globiformis T6.

The gene encoding an extracellular isomalto-dextranase, designated imd, was isolated from the chromosomal DNA of Arthrobacter globiformis T6 and cloned and expressed in Escherichia coli. A single open reading frame consisting of 1,926 base pairs that encoded a polypeptide composed of a signal peptide of 39 amino acids and a mature protein of 602 amino acids (M(r), 65,900) was found. The primary structure had no significant homology with the structures of any other reported carbohydrases, including two other dextranases. Transformed E. coli cells carrying the 2.3-kb fragment overproduced isomalto-dextranase into the periplasmic space under control of the promoter of the imd gene itself.

Amino Acid Sequence↗
PubMed1994

The sphR product, a two-component system response regulator protein, regulates phosphate assimilation in Synechococcus sp. strain PCC 7942 by binding to two sites upstream from the phoA promoter.

In the photosynthetic cyanobacterium Synechococcus sp. strain PCC 7942, the sphS and sphR genes were previously suggested to encode a typical pair of two-component signal transduction proteins. A deletion mutant strain lacking these genes failed to exhibit induction of alkaline phosphatase, the phoA gene product, in response to phosphate limitation in the medium. The SphR protein was overexpressed in Escherichia coli and then purified to near homogeneity. A truncated form of the SphS polypeptide (named SphS*) was also isolated. Here, we demonstrate that purified SphR is phosphorylated by phosphotransfer from SphS and binds to two distinct sites upstream from the phoA promoter. From these results, we conclude that the SphS and SphR proteins are directly involved in the regulation of phoA transcription in response to phosphate limitation in Synechococcus species.

Alkaline Phosphatase↗
PubMed1994

Expression of mRNA for natriuretic peptide receptor subtypes in bovine kidney.

The localization of mRNA for atrial natriuretic peptide (ANP) receptor subtypes (A, B, C) in the kidney was examined. Quantitative analysis of the ribonuclease protection assay showed that the numbers of type A receptor (ANPRA) mRNA were 6.9 x 10(7) in the glomeruli and 10.4 x 10(7) molecules/micrograms of total RNA in the inner medulla, and that of type C receptor (ANPRC) mRNA was 21.7 x 10(7) molecules/micrograms of total RNA in the glomeruli. The type B receptor (ANPRB) mRNA was present in smaller numbers (4.5-4.9 x 10(6) molecules/micrograms of total RNA) evenly throughout the kidney fractions. In situ hybridization demonstrated both ANPRA and ANPRC mRNA selectively in the glomerular epithelial cells and ANPRA mRNA in the collecting duct cells of the inner medulla. ANPRC was also localized on the foot processes of glomerular epithelial cells by immunohistochemistry using a specific antibody against the receptor. These results indicate that ANPRA is the major biologically active receptor for the ANP family of hormones in the kidney and is present selectively on the glomerular epithelial cells and inner medullary collecting duct cells. These cells are presumed to play a role in the regulation of glomerular filtration rate and sodium excretion induced by the family of ANP.

Animals↗
PubMed1994

Angiotensin II receptor antagonist TCV-116 induces regression of hypertensive left ventricular hypertrophy in vivo and inhibits the intracellular signaling pathway of stretch-mediated cardiomyocyte hypertrophy in vitro.

BACKGROUND: Previous studies have demonstrated that angiotensin II (Ang II) acts as a growth-promoting factor directly on cardiac myocytes and that angiotensin-converting enzyme inhibitor induces regression of hypertrophied hearts both in experimental animals and in humans. These results suggest that the renin-angiotensin system (RAS) is involved in the formation of left ventricular hypertrophy (LVH). To elucidate the role of RAS in the progression of cardiac hypertrophy, we evaluated the effect of an Ang II receptor antagonist on LVH in spontaneously hypertensive rats (SHRs) and investigated the molecular mechanisms by which antagonizing Ang II receptors reduces cell hypertrophy of myocytes using the in vitro model of mechanical stretch. METHODS AND RESULTS: In the in vivo study, we treated SHRs with the nonpeptide Ang II receptor antagonist TCV-116 (0.1, 1, or 10 mg/kg per day) or hydralazine (10 mg/kg per day). Blood pressure was measured by the tail-cuff method, and wall thickness of left ventricle was serially monitored using M-mode echocardiography. Rats were killed at the age of 13, 17, 21, or 25 weeks, and left ventricular (LV) weight, transverse diameter of cardiomyocytes, relative amount of V3 myosin heavy chain (MHC), and degree of interstitial collagen accumulation were examined. Untreated SHRs progressively developed severe hypertension, but treatment with TCV-116 or hydralazine inhibited the increase in blood pressure. Treatment with TCV-116 reduced LV weight, LV wall thickness, transverse diameter of myocytes, relative amount of V3 MHC, and interstitial fibrosis, whereas treatment with hydralazine slightly prevented an increase in LV wall thickness but did not exert significant reduction in other parameters. In the in vitro study, neonatal rat cardiomyocytes were cultured on deformable silicone dishes and mechanically stretched with or without pretreatment of CV-11974 (an active metabolite of TCV-116), and [3H]phenylalanine incorporation, activity of mitogen-activated protein (MAP) kinase, and c-fos mRNA expression were analyzed. Pretreatment of cultured cardiomyocytes with 10(-7) mol/L CV-11974 inhibited an increase in [3H]phenylalanine incorporation, MAP kinase activity, and c-fos gene expression induced by stretch of cardiomyocytes. CONCLUSIONS: The Ang II receptor antagonist TCV-116 induced regression of cardiac hypertrophy and had cardioprotective effects on hypertrophied myocardium in vivo, and antagonizing Ang II receptors inhibited intracellular signaling of stretch-mediated cardiomyocyte hypertrophy in vitro. These results suggest a crucial role of the cardiac RAS in the development of LVH produced by pressure overload.

Angiotensin II↗
PubMed1994

Importance of stereospecific positioning of the upstream cis-acting DNA element containing a curved DNA structure for the functioning of the Escherichia coli proV promoter.

The mechanism by which the Escherichia coli proV promoter is activated more than 100-fold in response to the medium osmolarity, without the help of any known trans-acting activators, is not yet fully understood. In this context, it has recently begun to be realized that structural features, not the primary sequences, of cis-acting DNA elements may be important for transcriptional regulation in prokaryotes. From this point of view, in this study the proV promoter was characterized by constructing a series of spacer-insertion mutants in a proV-lacZ fusion on the chromosome. Here it was found that the upstream cis-acting sequence must be positioned stereospecifically with respect to the principal -35 and -10 regions for the proV promoter to be fully activated. In this regard, it was suggested that an overall DNA structure, particularly DNA curvature, is an important cis-acting parameter for activation of the proV promoter.

Bacterial Proteins↗
PubMed1994

Antitumor polysaccharides from a Chinese mushroom, "yuhuangmo," the fruiting body of Pleurotus citrinopileatus.

A water-soluble polysaccharide, FI, and water-insoluble polysaccharides, FII and FIII were extracted from Pleurotus citrinopileatus mushrooms. After fractionation, the antitumor activity of the fractions against Sarcoma 180 implanted in mice were examined, and the following active polysaccharides were obtained: Water-soluble polysaccharides FIo: A heteropolysaccharide containing 9.8% protein, and composed of glucose, mannose, arabinose, and galactose. FA-2-b-beta: A glycoprotein consisting of glycan:protein = 40:60 w/w, with the glyco-chain composed of glucose, xylose, mannose, galactose, and fucose. FA-3: A glycoprotein consisting of glycan:protein = 50:50 w/w, and glycan moiety consisting of glucose, galactose, xylose, mannose, and fucose. Water-insoluble polysaccharides. FIII-1: Protein-containing beta-D-glucans, FIII-1-a, and -b were obtained from FIII-1 by gel filtration, composed of glucan:protein = 80:20, and 68:32 w/w, respectively. The glucan moieties of both were almost all (1-->3)-beta-D-glucan, and their molecular weights were 68 x 10(4) and 40 x 10(4). FIII-2: Protein-containing beta-D-glucans, FIII-2-a, and -b, were obtained from FIII-2 by gel filtration, with molecular weights 190 x 10(4) and 120 x 10(4), respectively. Both were almost all composed of glucan:protein = 87:13 w/w. Both glucan moieties were mainly (1-->3)-beta-D-glucan.

Amino Acids↗
PubMed1994

Antitumor active protein-containing glycans from the Chinese mushroom songshan lingzhi, Ganoderma tsugae mycelium.

A water-soluble polysaccharide, FI, extracted from the mycelium of Granoderma tsugae, was fractionated and purified by ion-exchange chromatography, gel filtration, and affinity chromatography. Sixteen polysaccharides obtained were examined for antitumor effects on Sarcoma 180 in mice. The three active polysaccharides obtained were as follows: FIo-a: A glycan-protein complex containing 9.3% protein, and having a hetero-glyco-chain of mannose and xylose. FIo-b-alpha: Molecular weight 10,000, glucan-protein complex containing 25.8% protein. The inhibition ratio was 61.8% against the solid cancer Sarcoma 180/mice; the survival ratio was more than 194% of the control group (100). FA-1-b-alpha: Molecular weight 16,000, a complex of glycan:protein = 42:58 w/w, consisting of glucose as a main component, and associated with arabinose, mannose, xylose, and galactose. This had a tumor inhibition ratio of 56% and a survival ratio of more than 182%. Comparison of active glycan with the fruiting body and mycelium: Among water-soluble polysaccharides of fruiting body, FIo-a and FA-1, with antitumor activity, were both glucogalactan-protein complexes of molecular weight 10,000, but that of mycelium was a homoglucan protein complex in FIo-b-alpha and heteroglucan protein in both FA-1-a and FA-1-b-alpha. The heteroglucan had a low tumor inhibition ratio, but caused a high survival ratio in mice.

Amino Acids↗
PubMed1994

Endoscopic endoprosthesis for large stones in the common bile duct.

Endoscopic biliary endoprosthesis was performed for 34 high-risk patients with common bile duct stones too large to be extracted by conventional endoscopic means. Bile duct drainage was established in all the patients without complications. Late complications developed in four patients and included cholangitis (three) and biliary pain (one). Twenty-five patients underwent a second endoscopic retrograde cholangiopancreatography between 4 and 30 months (mean, 15.1) during follow-up. Stone fragmentation was obtained in 76% (19/25) of the patients. Ten patients had complete stone clearance, and nine patients had disintegrated stones which could be readily removed endoscopically. The remaining nine patients were followed up with endoprostheses in situ for four to 60 months (mean, 24.8) without any symptoms. These results suggest that endoscopic endoprosthesis for difficult common bile duct stones is an effective method to clear the duct in selected cases, as well as an important definitive treatment in high-risk patients.

Aged↗
PubMed1994

[Induction of prostatic buds in the absence of androgens].

We have investigated the non-androgenic factors that induce the prostatic buds from the sinus epithelium. The buds were found to be induced in the explants cultured in the androgen-deficient medium containing 20 ng/ml rat keratinocyte growth factor (KGF) irrespective of the sex of the sinus.

Animals↗
PubMed1994

Aspects of morphogenesis and cytodifferentiation of endoderm offered by organ culture techniques.

In this article, I will summarize the results obtained in my laboratory from the study of organogenesis since 1970. The results have shown region-specific directive influence of mesenchymes and reactivity of endoderms including those of the allantois, yolk sac, digestive tract and urogenital sinus. It was also shown that morphogenesis can take place without coupling cytodifferentiation in heterologous recombination experiments.

Androgens↗
PubMed1994

The retinoblastoma binding factor 1 (RBF-1) site in RB gene promoter binds preferentially E4TF1, a member of the Ets transcription factors family.

The tumor suppressor retinoblastoma gene product, pRB, is a well known regulator of G1/S cell cycle progression. Moreover, mutational inactivations within the retinoblastoma gene (RB) are found in many human malignant tumors, and thus, believed to be an essential step in tumor formation. The human RB gene is considered as a housekeeping gene with no characteristic TATA or CAAT elements in its promoter region, but the sequence between 206 and 185 bases upstream of the initiation codon, essential for RB promoter activity, contains putative Sp1 and ATF recognition sites. We have previously reported that point mutations in this region, causing low penetrance retinoblastomas, completely reduced RB promoter activity, and that a nuclear factor, named RBF-1 (retinoblastoma binding factor 1), could specifically bind to this sequence, overlapping Sp1 recognition sequence. We show here, that RBF-1 can recognize a specific DNA sequence, 5'-GGCGGAAGT-3', overlapping the Sp1 and ATF sites and corresponding to the consensus DNA binding site for members of Ets transcription factors family. When RBF-1 site was used for sequence specific DNA affinity purification from erythroleukemia cells, reconstitution assays, immunoblotting analysis and peptide mapping show that the two major co-purified proteins are identical to human E4TF1-60 and -53 proteins. This reveals that E4TF1 can bind to the RBF-1 site of RB gene promoter, which, thus, constitutes a new target for this member of Ets transcription factors family.

Base Sequence↗
PubMed1993

Sequence of a fission yeast gene encoding a protein with extensive homology to eukaryotic elongation factor-1 gamma.

A polypeptide with an apparent molecular mass of 23 kDa was identified, that exhibited an affinity to a 491-bp DNA derived from one of the Schizosaccharomyces pombe centromeric DNAs (cen1). After determining its N-terminal amino acid (aa) sequence, a Sz. pombe genomic DNA encompassing the coding sequence of the isolated protein was cloned, and a 2.3-kb genomic DNA region sequenced. Further sequence analysis of cDNA clones, originating from this particular genomic region, confirmed the existence of an open reading frame with a short intron, which encodes a 409-aa protein with striking homology to eukaryotic elongation factor-1 gamma.

Amino Acid Sequence↗
PubMed1993

Distribution of bent DNA structures in the fission yeast centromere.

To gain a clue as to the functional significance of DNA curvature, we experimentally characterized the distribution of bent DNA structures throughout the 35-kb cen1 sequence, one of the isolated functional centromeric DNA of the fission yeast, Schizosaccharomyces pombe. It was revealed that a relatively large central portion of cen1, covering a 2.2-kb DNA sequence, displays a remarkable DNA curvature.

Centromere↗
PubMed1993

The cyanobacterium, Synechococcus sp. PCC7942, possesses two distinct genes encoding cation-transporting P-type ATPases.

P-type (or E1 E2-type) ATPases comprise a large family of prokaryotic and eukaryotic proteins capable of transporting a variety of cations, and function in a wide variety of cellular processes. The present study was carried out to search for genes encoding P-type ATPases in the phototrophic cyanobacterium, Synechococcus sp. PCC7942. We succeeded in cloning two genes each encoding P-type ATPases from this bacterium. It was found that Synechococcus at least, two distinct P-type ATPases; one belongs to the family of typical prokaryotic P-type ATPases and the other markedly resembles eukaryotic P-type ATPases. An insertion mutant lacking either of these two ATPase-genes was constructed. The results showed that the growth of these mutants is hypersensitive to osmotic stress upon addition of NaCl or sorbitol to the medium.

Adenosine Triphosphatases↗
PubMed1993

Cloning of a sensory-kinase-encoding gene that belongs to the two-component regulatory family from the cyanobacterium Synechococcus sp. PCC7942.

A screening method employing Escherichia coli was adopted to clone a sensory-kinase (SK)-encoding gene directly from a phylogenetically distant species, the phototrophic cyanobacterium Synechococcus sp. PCC7942. From the Synechococcus chromosomal DNA, we searched for DNA clones which are able to complement phenotypically not only an E. coli envZ mutant for the expression of ompC, but also an E. coli phoR/creC mutant for the expression of alkaline phosphatase. These E. coli genes are known to encode SK. A 0.75-kb DNA fragment was thus cloned under the control of the E. coli lac promoter carried on an E. coli plasmid vector. A larger DNA fragment encompassing an entire open reading frame was then cloned and its complete nucleotide (nt) sequence determined. The nt sequence corresponds to a gene that encodes a 43,280-Da protein of 387 amino acids with a high degree of homology to the bacterial SK. Thus, we succeeded in cloning a SK-encoding gene, which most likely functions in signal transduction in Synechococcus sp. PCC7942. Hence, the gene was designated sasA (Synechococcus adaptive-response SK A). The purified SasA protein was demonstrated in vitro to undergo autophosphorylation.

Adaptation, Physiological↗
PubMed1993

In vitro irradiation is able to cause RET oncogene rearrangement.

Elevated risk of thyroid cancers among the atomic bomb survivors as compared to the nonexposed population suggests that some genetic events related to thyroid cancer must be caused by ionizing radiation. Accordingly, inducibility of RET oncogene rearrangements, i.e., the generation of the RET-PTC oncogene, specific for thyroid cancer, was investigated among human undifferentiated thyroid carcinoma cells (8505C), which do not have RET oncogene rearrangement, after 0, 10, 50, and 100 Gy of in vitro X-irradiation by means of reverse transcription polymerase chain reaction. After testing 10(8) cells at each dose point, 3 independent samples obtained with 50 Gy of X-irradiation and 6 independent samples obtained with 100 Gy of X-irradiation showed a rearranged RET oncogene amplified band. No rearranged transcripts were obtained from cells irradiated with 0 or 10 Gy. All of the transcripts were sequenced and found to contain the D10S170 and RET sequence. Interestingly, two types of rearrangements were included in these transcripts: one is specific for thyroid cancer and the other, which contains a 150-base pair insert, is atypical, not usually seen in vivo. This insert was found to be the exon of D10S170. Furthermore, in fibrosarcoma cells (HT1080), X-irradiation also induced RET oncogene rearrangements, which included the same two types of rearrangements observed in the X-irradiated thyroid cells (8505C). These results are in favor of the hypothesis that some radiation-induced thyroid cancers, including those among atomic bomb survivors, might have developed when a growth advantage was obtained through a specific form of RET oncogene rearrangement induced by radiation exposure.

Aged↗
PubMed1993

A variant form of the type C atrial natriuretic peptide receptor generated by alternative RNA splicing.

A novel form of type C atrial natriuretic peptide receptor (ANP-C) was identified, characterized, and shown to be a product of alternative RNA splicing. Sequencing of several clones of ANP-C cDNA isolated from a bovine lung cDNA library revealed the presence of a new clone encoding a 536-amino acid ANP-C. The nucleotide sequence of this novel clone is very similar to that of the previously cloned ANP-C which consists of 537 amino acids. The sole difference between the two clones is the presence or absence of a very short segment of only 3 base pairs (bp), by which the sequence Ser472-Gly473 of the longer form is changed to Cys472 in the variant form; the other regions are identical. Alignment of the cDNA sequences with that of the ANP-C gene indicated that the two forms of ANP-C receptor arise from a single gene by alternative splicing using one donor and two closely located acceptor sites. Biochemical and pharmacological characterization using a mammalian cDNA expression system indicated that the two variants are similar in their subunit structures, in ligand-binding properties, and even in internalization kinetics. Polymerase chain reaction amplification of the transcripts demonstrated that 1) although the novel form is a minor species, it is present in various tissues such as the lung, kidney, adrenal, and vascular smooth muscle and endothelial cells and 2) the ratio of the novel variant form to the original longer form remains almost constant in these tissues. The alternative splicing reported here contributes to the diversity of ANP receptors; however, its physiological significance remains to be clarified.

Affinity Labels↗