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Biomedical subjects

T Moriuchi

Publications and source records attributed to T Moriuchi.

At least 73 records · Page 4Linked to original sources
PubMed1991

Structural analysis of the human HOX4A homeobox gene.

The HOX4A gene, one of a cluster of homeobox-containing genes on human chromosome 2, has been isolated by screening a genomic cosmid library with the HOX4B cDNA probe. The amino acid sequence was predicted according to the conceptual translation of 13 homology groups of human HOX genes (1). The HOX4A gene consists of at least two exons separated by a long intron of 1860 bp. The HOX4A protein predicted from the nucleotide sequence of the HOX4A gene is comprised of 416 amino acid residues. Comparison of the predicted HOX4A protein with the HOX2G protein revealed three regions of sequence similarity: an N-terminal octapeptide, a hexapeptide (pre-box) upstream of the homeodomain, and the homeodomain at the C-terminus.

Animals↗
PubMed1991

Molecular cloning of human plasma glutathione peroxidase gene and its expression in the kidney.

A genomic clone containing the human plasma glutathione peroxidase (GSH-Px) gene has been isolated using a rat plasma GSH-Px cDNA as a probe. The partial nucleotide sequence of the clone completely matched the sequence of the human plasma GSH-Px cDNA. The results of Southern blot hybridization indicate that the human plasma GSH-Px gene consists of at least 4 exons and 3 introns, and spans about 12 kb. RNA blot analysis demonstrated that the human plasma GSH-Px gene is expressed in the kidney.

Animals↗
PubMed1991

Autopsy study on the weight of the heart, liver, kidney and brain in Duchenne muscular dystrophy.

The weight of the major internal organs in 188 autopsy cases of Duchenne muscular dystrophy were studied in relation to the clinical course of disease and age of the patients at death. The hearts were classified into three groups (atrophic, normal weight and hypertrophic), each constituting about one-third of the total cases. Cardiac atrophy was most frequent in the older cases of more than 20 years of age, whereas hypertrophy was more frequent in younger cases with the highest incidence in cases under 15 years. The liver showed marked atrophy in most cases (144 of 178 cases). The incidence of liver atrophy increased with the age of the patients and all 55 patients of over 20 years of age had an abnormally small liver. These marked atrophies of the heart and liver appeared to have occurred in parallel with loss of body weight due to skeletal muscle wasting. Myocardial dystrophy might be another causative factor for cardiac atrophy. The myocardial hypertrophy seemed to be caused by an intercurrent congestive failure. Renal atrophy was observed in only a few of the extremely emaciated patients. The brain weight was within the normal range in most cases.

Adolescent↗
PubMed1990

Immunohistochemical study of fiber types in human extraocular muscles.

Fiber types in human extraocular muscle (h-EOM) were examined immunohistochemically with antibodies against slow tonic (anti-ALD) and slow twitch (anti-SOL) myosins. Four types of muscle fiber in h-EOM were distinguishable according to their reactivities with these antibodies. Groups 1 and 2 fibers reacted with both antibodies, group 1 fibers showing stronger reactivity than group 2 fibers with anti-ALD. Group 3 fibers reacted only with anti-SOL. Group 4 fibers did not react with either antibody. The latter were the most common, and were the main fibers in both the peripheral (outer orbital) and central zones of h-EOM. The next most common were group 1 fibers, which were located mainly in the peripheral layer. Group 2 fibers were less common, but were the second most common type in the central layer. Group 3 fibers were only minor constituents. Multiple innervations were observed in some fibers of groups 1 and 2, and group 1 fibers were suggested to be slow tonic myofibers in h-EOM. These specific immunohistochemical and physiological features of h-EOM seem to be the basis of the low morbidity seen in the usual types of muscular dystrophy.

Aged↗
PubMed1990

Drosophila proliferating cell nuclear antigen (cyclin) gene: structure, expression during development, and specific binding of homeodomain proteins to its 5'-flanking region.

The genomic and cDNA clones for a Drosophila melanogaster proliferating cell nuclear antigen (PCNA) (cyclin) were isolated and sequenced. The coding sequence for a 260-amino-acid residue polypeptide was interrupted by a single short intron of 60 base pairs (bp), and about 70% of the deduced amino acid sequence of the Drosophila PCNA was identical to the rat and human PCNA polypeptides, with conserved unique repeats of leucine in the C-terminal region. Genomic Southern blot hybridization analysis indicates the presence of a single gene for PCNA per genome. The PCNA mRNA was detected at a high level in adult ovaries, unfertilized eggs, and early embryos and at low levels in the other developmental stages. The major transcription initiation site (cap site) was localized at 89 bp upstream from the ATG codon. Neither a TATA box nor a CAAT box was found within the 600-bp region upstream of the cap site. Clusters of 10 bp of sequence similar to the binding sites for Drosophila proteins containing homeodomains were found in the region from -127 to -413. DNase I footprint analysis revealed that the Drosophila homeodomain proteins coded by even-skipped and zerknüllt genes can specifically bind to these sites. These results suggest that the expression of the PCNA gene is under the control of genes coding for homeodomain proteins.

Amino Acid Sequence↗
PubMed1990

Molecular cloning of cDNA coding for rat plasma glutathione peroxidase.

The plasma glutathione peroxidase (PGSH-PO), which is different from erythrocyte glutathione peroxidase (EGSH-PO) in immunochemical property and substrate specificity, was purified from male Wistar rat serum. The amino acid sequence of 5 independent peptides were determined and a cDNA clone for this enzyme was isolated from placental cDNA library. The nucleotide sequence of the cDNA revealed that, similar to EGSH-PO cDNA, the seleno-cysteine was genetically encoded by "TGA" codon. On comparing the nucleotide sequences of EGSH-PO and PGSH-PO, no significant homology was found in the vicinities of "TGA" codons of both enzymes.

Amino Acid Sequence↗
PubMed1989

Gene for proliferating-cell nuclear antigen (DNA polymerase delta auxiliary protein) is present in both mammalian and higher plant genomes.

Proliferating-cell nuclear antigen (PCNA; also called cyclin) was originally described in proliferating mammalian cells as a nuclear protein with an apparent Mr of 33,000-36,000 and recently was found to be a DNA polymerase delta auxiliary protein. To elucidate whether PCNA/cyclin is a universal protein necessary for proliferation of eukaryotes, a search was conducted for PCNA/cyclin homologues in higher plants. In Southern blot-hybridization analysis, a rat PCNA/cyclin cDNA probe hybridized with homologous sequences in genomic DNAs from rice, soybean, and tobacco. A PCNA/cyclin-related molecular clone (pCJ-1) was isolated from rice DNA and was partially sequenced. The pCJ-1 probe hybridized with a 1.2-kilobase transcript in RNA from rice root tips and shoots. Immunoblot analysis of the soluble extract of soybean root tips with monospecific anti-PCNA/cyclin identified an immunoreactive protein with an apparent Mr of 34,000. Immunohistochemical analysis revealed the presence of an immunoreactive PCNA/cyclin protein in the nuclei of cells in the meristem of soybean root tips. The highly homologous nature of the gene for PCNA/cyclin throughout the animal and plant kingdoms suggests that the product of the gene plays an essential role in DNA replication in eukaryotes.

Amino Acid Sequence↗
PubMed1989

A polymorphic DNA marker genetically linked to congenital malformations.

Unusual restriction fragments were detected by DNA blot hybridization with PCNA (DNA polymerase-delta auxiliary protein) probe in one of seven cases of congenital malformations. Chromosomal in situ hybridization localized PCNA gene to region q31-35 of human chromosome 2. To discover the locus more closely associated with congenital malformations, a cloned DNA segment which has been mapped to chromosomal region 2q33-36 was tested for restriction fragment length polymorphisms (RFLPs) in these patients. The 2q33-36 probe hybridized with 2.1-kb, 1.9-kb and 1.7-kb fragments in ten normal control samples. In seven cases of congenital malformations examined, however, the band of 2.1 kb is absent in six cases and the band of 1.7 kb in one case. These results indicate that the locus closely linked to congenital malformations is present in the proximity of PCNA locus.

Child↗
PubMed1988

Localization in situ of c-myc mRNA and c-myc protein in adult mouse testis.

It has been suggested that c-myc, one of the proto-oncogenes, plays a role in normal somatic cell proliferation and differentiation. To define whether c-myc is only expressed during somatic cell division or is also expressed during meiotic cell division, the production of c-myc mRNA and protein were investigated in the mouse testis by using in situ hybridization with non-radioactive DNA probes and enzyme immunohistochemistry respectively. For in situ hybridization, T-T dimerized DNA probes were used and DNAs hybridized in situ were detected immunohistochemically using specific antibody against T-T dimer. The results indicate that c-myc mRNA and protein are expressed in a cell-cycle-dependent manner only in spermatogonia and not in spermatocytes and spermatids.

Animals↗
PubMed1988

Use of non-radioactive DNA probes for the characterization of adult T-cell leukemia cells.

DNA fragments were labeled with dinitrophenyl (DNP) residues by the reaction with 2,4-dinitrobenzaldehyde in alkaline condition and the labeled DNA was used as a probe for non-radioactive in situ hybridization. DNP-labeled DNA probes for T cell receptor beta chain, c-myc and HTLV-1 were hybridized in situ to mRNA on cell specimens fixed with Carnoy's fixative. DNA-mRNA hybrids were detected immunohistochemically using anti-DNP antibodies. Cytoplasms of adult T cell leukemia cells were stained with varied intensity when these probes were used. More than 70% of cells were positively stained with T cell receptor probe. However, less than 30% of cells were stained with c-myc and HTLV-1 probes. The present study indicates that non-radioactive in situ hybridization can be used for the characterization and classification of leukemia.

Animals↗
PubMed1987

Molecular cloning of cDNA coding for rat proliferating cell nuclear antigen (PCNA)/cyclin.

The 'proliferating cell nuclear antigen' (PCNA), also known as cyclin, appears at the G1/S boundary in the cell cycle. Because of its possible relationship with cell proliferation, PCNA/cyclin has been receiving attention. PCNA/cyclin is a non-histone acidic nuclear protein with an apparent mol. wt of 33000-36000. The amino acid composition and the sequence of the first 25 amino acids of rabbit PCNA/cyclin are known. Using an oligonucleotide probe corresponding to the sequence of the first five amino acids, a cDNA clone for PCNA/cyclin was isolated from rat thymocyte cDNA library. The cDNA (1195 bases) contains an open reading frame of 813 nucleotides coding for 261 amino acids. The 3'-non-coding region is 312 nucleotides long and contains three putative polyadenylation signals. The mol. wt of rat PCNA/cyclin was calculated to be 28 748. The deduced amino acid sequence and composition of rat PCNA/cyclin are in excellent agreement with the published data. Using the cDNA probe, two species of mRNA (1.1 and 0.98 kb) were detected in rat thymocyte RNA. Southern blot analysis of total human genomic DNA suggests that there is a single gene coding for PCNA/cyclin. The deduced amino acid sequence of rat PCNA/cyclin has a similarity with that of herpes simplex virus type-1 DNA binding protein.

Amino Acid Sequence↗
PubMed1986

Polymorphism of HLA-DR beta chains in DR4, -7, and -9 haplotypes: implications for the mechanisms of allelic variation.

We have isolated and sequenced cDNA clones corresponding to the DR beta 1 and DR beta 2 loci from two homozygous B-cell lines typed as DR7 (Burkhart) and DR9 (ISK). These nucleotide sequences were compared to beta 1 and beta 2 chains of other DR haplotypes. The first-domain sequences of beta 2 chains are identical in DR4 and DR7 haplotypes. In addition, there is strong sequence homology within the 3' untranslated regions of beta 1 genes from DR4, -7, and -9 haplotypes, thus confirming the close evolutionary relationship among these three haplotypes. In contrast, the first-domain sequences of beta 1 molecules from these haplotypes are very different from each other and do not reflect the DR4, -7, -9 family relationship. Two explanations for the differences in degree of diversity between beta 1 and beta 2 chains are suggested. The differences may be a consequence of selection pressures; this implies functional differences for products of the beta 1 and beta 2 loci. Alternatively, closely linked segments of the human class II region may differ in their underlying rates of variation, independent of selection pressures, and this may in part account for the extraordinary diversity found in the beta 1 first domain.

Alleles↗
PubMed1986

Molecular cloning and nucleotide sequence analysis of rat PCNA/cyclin cDNA.

The 'proliferating cell nuclear antigen' (PCNA), also known as cyclin, accumulates in the nuclei of dividing and transformed cells and reacts with autoantibodies from certain lupus patients. A full-length cDNA (1195 bp) clone encoding PCNA/cyclin was isolated from rat thymocyte cDNA library. The nucleotide sequence reveals an open reading frame of 783 nucleotides coding for 28.7 kD protein. The predicted amino acid sequence and composition are in excellent agreement with the published protein data of rabbit PCNA. We report the entire nucleotide sequence of the cDNA and complete amino acid sequence for rat PCNA/cyclin.

Amino Acid Sequence↗
PubMed1986

[Enzyme-immuno-histo in situ hybridization].

In order to understand the physiological functions of normal as well as neoplastic cells, it is best to investigate at the level of individual cell. We have been involved for the past several years with the development of a method for the localization of mRNA in individual cells using hapten labeled cDNA. Specifically, cDNA are labeled with dinitrophenyl (DNP) and the labeled cDNA are hybridized with mRNA in cells. Then the hybridized DNP-cDNA are then localized immunohistochemically using peroxidase-labeled antibodies. In this investigation, we concentrated on establishing the best condition for the removal of proteins in order to expose mRNA and for the removal of non-specific back ground staining. It was found that some proteins treatment was necessary for the exposure of mRNA for well fixed tissues and that high concentration of formamide and low concentration of salt removed effectively the nonspecific staining. Using the backscattered electron imaging for by scanning electron microscope, specific mRNA was localized in cells and tissues at the ultrastructural level.

DNA↗