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Biomedical subjects

T Fujiwara

Publications and source records attributed to T Fujiwara.

At least 523 records · Page 29Linked to original sources

Cloning, expression pattern and mapping to 12p 13.2 --> p13.1 of CLAPS3, a gene encoding a novel clathrin-adaptor small chain.

From a human fetal-brain cDNA library we isolated a novel gene encoding a peptide homologous to clathrin-adaptor small chains in rat, mouse, and yeast. The cDNA, designated CLAPS3 (clathrin-associated/assembly/adaptor protein, small 3, 22 kDa), contained an open reading frame of 579 nucleotides encoding 193 amino acids. Northern-blot analysis revealed expression of a 1.35-kb transcript in all human tissues examined. This gene was mapped to chromosome bands 12p13.2 --> p13.1 by FISH.

Adaptor Protein Complex 3↗

Isolation and mapping of a human gene (DIFF6) homologous to yeast CDC3, CDC10, CDC11, and CDC12, and mouse Diff6.

We have isolated a novel human cDNA that encodes a protein homologous to murine H5 and Diff6, and to yeast CDC10, and mapped it to chromosome region 2q37 by fluorescent in situ hybridization. Its transcript has an open reading frame of 1,218 nucleotides encoding 406 amino acids. The deduced peptide sequence contained conserved domains rich in basic residues, GXXGXGKS--DXXG--TKXD, a motif of the GTPase superfamily. Different polyA sites accounted for generation of two transcripts. The major type, 3.5 kb long, was expressed ubiquitously in all human tissues examined, but a 2.0-kb alternative transcript lacking any long AU-rich element in the 3' non-coding region was expressed abundantly only in testis, heart and skeletal muscle.

Adult↗

Cloning, expression and chromosome mapping of adducin-like 70 (ADDL), a human cDNA highly homologous to human erythrocyte adducin.

From a human fetal-brain cDNA library we isolated a novel human cDNA, termed human adducin-like 70 (gene symbol ADDL), whose predicted amino acid sequence showed a high degree of homology to adducins. This cDNA clone (ADDL), which contained an open reading frame of 2,022 nucleotides encoding 674 amino acids, revealed 54%, 53%, and 59% identity in predicted amino acid sequence with alpha and beta components of human adducin and rat adducin 63, respectively. Human adducin-like 70 is likely to play an important role in the skeletal organization of the cell membrane. Northern blot analysis indicated ubiquitous expression of this gene in adult human tissues. We localized the gene to chromosome bands 10q24.2-->q24.3 by fluorescence in situ hybridization (FISH).

Adult↗

Isolation and mapping of the human glycoprotein M6 gene (GPM6A) to 4q33-->q34.

We have isolated a human cDNA that is highly homologous to the murine gene (Gpm6) encoding a membrane glycoprotein, M6. The human gene (GPM6A) contains an open reading frame of 834 nucleotides encoding a peptide of 278 amino acids. Northern-blot analysis revealed specific expression in human brain. We assigned the GPM6A locus to chromosome bands 4q33-->q34 by radiation hybrid mapping.

Amino Acid Sequence↗

Molecular cloning of UBE2G, encoding a human skeletal muscle-specific ubiquitin-conjugating enzyme homologous to UBC7 of C. elegans.

From a human fetal-brain cDNA library we isolated a novel ubiquitin-conjugating enzyme. The cDNA, designated UBE2G, contained an open reading frame of 510 nucleotides encoding 170 amino acids. The predicted peptide product showed 74% identity at the amino acid level with UBC7 of C. elegans, a high degree of homology with UBC7s of other species, and significant homologies with other subgroups of UBCs. Northern-blot analysis revealed strong expression of 4.4-kb, 2.4-kb, and 1.6-kb transcripts in skeletal muscle. Weak expression was observed in 15 other tissues examined. Radiation-hybrid mapping localized this cDNA to chromosome band lq42.

Animals↗

Isolation, characterization and chromosomal assignment of the human WNT7A gene.

The Wnt genes compose a large gene family encoding a group of secreted signaling molecules that have been implicated in oncogenesis and a number of developmental processes. We have isolated a full-length human cDNA clone that we consider to be a novel member of the Wnt gene family. The gene (WNT7A) encodes a deduced 349-amino-acid peptide with 98% identity in amino acid sequence to murine Wnt7a. Expression of this gene is restricted to certain tissues: placenta, kidney, testis, uterus, fetal lung, and fetal and adult brain. Furthermore, we have isolated a genomic clone of WNT7A and mapped it to chromosome 3p25 by fluorescent in situ hybridization.

Amino Acid Sequence↗

Isolation and mapping of RAB2L, a human cDNA that encodes a protein homologous to RalGDS.

We have isolated a novel human cDNA, designated RAB2L, that encodes a protein homologous to Ral guanine nucleotide dissociation stimulator (RalGDS). The predicted sequence of 122 amino acids revealed 39.5% identity with the C-terminal region of human RalGDS. As the C-terminal region of RalGDS is considered to interact with RAS, the product of this novel gene may also play some role in the ras-mediated signal transduction pathway. Northern analysis revealed that the gene RAB2L is expressed in all human tissues examined. We assigned this gene locus to chromosome band 6p21.3 by fluorescence in situ hybridization.

Amino Acid Sequence↗

Cloning, expression pattern and mapping to Xq of NAP1L3, a gene encoding a peptide homologous to human and yeast nucleosome assembly proteins.

From a human fetal brain cDNA library, we isolated a novel gene sharing significant homology with the genes of nucleosome assembly proteins (NAPs). This cDNA clone, designated NAP1L3 (nucleosome assembly protein 1-like 3), contained an open reading frame of 1518 nucleotides encoding 506 amino acids. Its predicted amino acid sequence showed 46% identity and 65% similarity with NAP1L. In its C-terminal half NAP1L3 contained several characteristic motifs strictly conserved with NAP1L and yeast NAP1, but the N-terminal half showed little conservation. Northern-blot analysis revealed strong expression of a 3.0-kb transcript in human adult brain and weak expression in heart. NAP1L3 was closely linked to a marker (DXS990) mapped to chromosome bands Xq21.3-->q22, where genes responsible for several X-linked mental retardation syndromes have been localized.

Amino Acid Sequence↗

Endothelin-1 and its receptor in hypertrophic cardiomyopathy.

Endothelin-1, a potent vasoconstrictor produced by vascular endothelial cells, activates the hypertrophic program in cultured heart muscle cells. However, the role of endothelin-1 in cardiac hypertrophy in humans is unknown. Therefore, we studied hypertrophic cardiomyopathy patients with normal pulmonary arterial pressure, in whom cardiac hypertrophy is a specific feature of the disease. Radioimmunoassay with a monoclonal antibody to human endothelin-1 showed that the plasma level of immunoreactive endothelin was more than twofold higher in hypertrophic cardiomyopathy patients than in control subjects (P < .005). In situ hybridization analysis of endomyocardial biopsy specimens showed positive signals of endothelin-1 type A receptor mRNA in ventricular myocytes of all specimens. The receptor expression in ventricular myocytes was similar between hypertrophic cardiomyopathy patients and control subjects. We propose that endothelin-1 might represent an important factor involved in hypertrophic cardiomyopathy. Whether endothelin-1 plays a causal role in cardiac hypertrophy or is a marker of its occurrence needs to be clarified.

Analysis of Variance↗

Factors that affect calorie-sensitive and calorie-insensitive reduction in blood pressure during short-term calorie restriction in overweight hypertensive women.

This study was performed to elucidate the factors that affect the reduction in blood pressure produced by calorie restriction in overweight women with essential hypertension. Fifty-one subjects were admitted to the metabolic ward of the hospital. After being fed a standard diet (6.3 to 8.4 MJ/d) for 2 weeks, the calorie-restricted group (n=34) was fed a low-calorie diet (1.9 MJ/d) for an additional 2 weeks. The calorie-nonrestricted group (n=17) was fed the standard diet for 4 weeks. Sodium and potassium intake was kept constant, as was the level of exercise activity. The calorie-restricted group was subdivided into "calorie-sensitive" and "calorie-insensitive" groups based on an average 5 mm Hg reduction in mean blood pressure during the low-calorie diet. The mean age was 51+/-6 years (mean+/-SD) in the calorie-sensitive group (n=16), which was significantly lower than the mean age of 61+/-6 years in the calorie-insensitive group (n=18). Multiple regression analysis indicated that age and change in body weight exhibited significant correlations with blood pressure reduction produced by calorie restriction among 17 parameters. Findings suggest that age can predict the extent of blood pressure reduction that would be obtained during 2 weeks of calorie restriction in overweight hypertensive women. The reduction in blood pressure may be related in part to the amount of weight loss.

Adult↗

Identification and characterization of a 230-kDa Golgi-associated protein recognized by autoantibodies from a patient with HBV hepatitis.

A serum from a patient with HBV hepatitis was found to contain autoantibodies reacting with various mammalian cells. Immunofluorescence staining of cultured cells with the autoantibodies revealed that the antigen was localized at perinuclear regions, where the Golgi markers alpha-mannosidase II and beta-COP were colocalized. The autoantigen disappeared from the perinuclear regions upon incubation with the fungal metabolite brefeldin A, and the immunostainable structures were fragmented into vesicles by treatment with nocodazole. These results strongly indicate that the antigen is localized at the Golgi complex. Immunoblots of cell lysates showed that the autoantibodies recognized a single protein with a molecular mass of 230 kDa in a variety of cell lines, indicating that the 230-kDa antigen is a conserved protein among mammalian species. We designated this protein GCP230 (Golgi complex-associated protein with a molecular mass of 230 kDa). when a postnuclear fraction was prepared and centrifuged, GCP230 was recovered in both cytosol and membrane fractions. Peripheral interaction of GCP230 with membranes was confirmed by phase separation in Triton X-114 solution and by extraction with sodium carbonate. Taken together, these results indicate that GCP230 is a peripheral membrane protein of the Golgi derived from the cytosol, although its function is not known at present.

Autoantibodies↗

The surface properties of chemically synthesized peptides analogous to human pulmonary surfactant protein SP-C.

A number of peptides with different lengths corresponding to various regions of human pulmonary surfactant protein SP-C were synthesized and their activity evaluated to improve in vitro surface activities and in situ lung pressure-volume characteristics of a ternary lipid mixture composed of dipalmitoylphosphatidylcholine, phosphatidylglycerol and palmitic acid (75:25:10, w/w). SP-C (1-35), a synthetic peptide with the entire length of human SP-C, and some other peptides with various lengths of its partial sequences were remarkably active. All of these peptides shared a common core sequence of (C)CPVHLKRLLIVVVVVVLIVVVIVGAL(L). Any deletion in this core sequence resulted in reduction of activity of the peptide. SP-C (5-31) and SP-C (6-32), the minimum peptides containing the core sequence, were combined with the ternary lipid mixture at the final peptide concentration of 2% (w/w) into synthetic surfactants which showed excellent properties comparable with those of Surfacten, a commercially available modified bovine lung surfactant. In a Langmuir-Wilhelmy surface balance, the synthetic surfactant containing SP-C (6-32) spread and adsorbed quickly to reach a surface tension of 30.8 mN/m at 30-s spreading time and 41.2 mN/m at 1-min adsorption time, respectively; the presence of SP-C (6-32) significantly prevented the decrease of surface activity of the ternary lipid mixture during dynamic compression-expansion cycles. Furthermore, tracheal instillation of the synthetic surfactant containing SP-C (6-32) at the dose of 50 mg of phospholipids/kg improved lung pressure-volume characteristics of immature rabbit neonates to a level similar to that of mature neonates at term.

Amino Acid Sequence↗

Noradrenergic hyperinnervation in the heart of stroke-prone spontaneously hypertensive rats (SHRSP).

Noradrenergic (NA) nerve fiber density was investigated in the subepicardium and myocardium of ventricles in stroke-prone spontaneously hypertensive rats (SHRSP) and was compared with that of normotensive Wistar Kyoto (WKY) rats. Fluorescent NA nerve fibers in the subepicardium of the right and left ventricles of both strains at the ages of 10, 30, 60, 90, and 180 days were examined by the glyoxylic acid method. NA nerve fibers in the myocardium of the right and left ventricles and the ventricular septum of both strains at the ages of 30, 90, and 180 days were also examined in a similar manner. The density of NA nerve fibers was measured by quantitative image analysis. The distribution pattern of NA nerve fibers in the entire subepicardium of ventricles of both strains showed a meshwork pattern throughout the examination period. In sections of the myocardium, NA nerve fibers were distributed between heart muscle cells and around blood vessels in both strains at all ages examined. The densities of NA nerve fibers in the subepicardium of the ventricles of SHRSP were significantly higher than those of WKY rats at all ages examined except for the subepicardium of the left ventricle at 90 days of age. The densities in the myocardium of the right ventricle in 30- and 90-day-old SHRSP were significantly higher than those in WKY rats. The ratios of NA nerve fiber density of SHRSP to that of WKY rats were greater in the subepicardium of the right and left ventricles, except at 90 days of age, and in the myocardium of the right ventricle of younger animals as compared with older ones. NA hyperinnervation in the subepicardium and myocardium of the ventricles of SHRSP may be a primary change of the heart before the onset of hypertension and may be caused by hyperfunction of the stellate ganglia.

Adrenergic Fibers↗

Long-term results of emergency coronary artery bypass grafting.

During a 20-year period, 364 patients underwent coronary artery bypass grafting (CABG) for the treatment of ischemic heart disease. Among these patients, 28 underwent emergency surgery. The reasons for performing emergency CABG were unstable angina in 15 patients, impending myocardial infarction in 12 patients and congestive heart failure in 1 patient. Eleven patients died postoperatively. Eight variables were examined by univariate analysis for their influence on the occurrence of a hospital death. Lack of a history of myocardial infarction, intraaortic balloon pumping (IABP) and acute coronary occlusion were all found to be predictors of hospital death. Seventeen patients were followed up for 12 years. There was no cardiac death and actuarial survival at 12 years was 63%. The 14 survivors are now in NYHA functional class I or II. Although the operative mortality rate is high after emergency CABG, a fair prognosis can be expected if the patients survive surgery.

Adult↗

Effect of low dose cyclophosphamide on the synthesis of acute phase protein and its significance for cancer chemotherapy.

Patients with far advanced colorectal cancers received chemotherapy consisting of low-dose cyclophosphamide (LDCY) 333 mg/m2 every four weeks intravenously and by oral administration of 5'-DFUR (a masked compound of 5-Fluorouracil). Serum levels of immunosuppressive acidic protein (IAP), an acute phase protein, were measured every four weeks for a total of thirty-one LDCY trials of ten patients. LDCY chemotherapy significantly decreased the IAP levels in cancer patients with high IAP levels. These results suggested that LDCY chemotherapy could counteract host responses against tumors and could have decreased immunosuppressive responses in cancer patients.

Acute-Phase Proteins↗