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Biomedical subjects

H Fujii

Publications and source records attributed to H Fujii.

At least 613 records · Page 34Linked to original sources

Cloning of a cDNA encoding rat intestinal 15 kDa protein and its tissue distribution.

A cDNA encoding rat intestinal 15 kDa protein was isolated and sequenced from a rat ileum-specific cDNA library. This cDNA was found to contain an open reading frame of 384 nucleotides as well as 5' (27 nucleotides) and 3' (46 nucleotides) non-coding regions. The deduced sequence of 127 amino acids was identical to that of rat I-15P which was purified from rat intestinal epithelium. The nucleotide sequence of the open reading frame exhibited 79% identity to that of the porcine gastrotropin. Northern blot analysis indicated that the same size of transcript as that of the ileum was detected in the ovary, suggesting that I-15P or a homologous protein might be involved in the metabolism of steroids in steroid hormone-producing tissues.

Amino Acid Sequence↗

Heterogeneity of carbonic anhydrase and 68 kDa neurofilament in nerve roots analyzed by two-dimensional electrophoresis.

The heterogeneity of carbonic anhydrase (CA) and 68 kDa neurofilament (NF) in nerve roots of Japanese monkeys (Macaca Macacus) was analyzed by two-dimensional electrophoresis combined with immunoblotting. CA was detected as heterogeneous spots of type I and II having the same molecular weight (M(r)) but different isoelectric point (pI), in the range of pH 4.5-5.5. CA had much higher concentrations in soluble fractions of dorsal roots than ventral roots but not in membrane-bound fractions of either root. NF was detected as a single spot at the position of 68 kDa and pI 5.2 in soluble fractions of both roots, however, it was detected in the membrane-bound fractions as heterogeneous spots having different molecular masses and pI in the range of 60-68 kDa and 5.0-5.5, respectively. The presence of heterogeneous CA type I and II at high concentrations in dorsal roots enables us to distinguish the sensory nerves from the motor nerves. Furthermore, the heterogeneity of NF in membrane-bound fractions gives us information for understanding the dynamic assembly and the degradation of NF in nerve roots.

Animals↗

Hybrid cell extinction and re-expression of Oct-3 function correlates with differentiation potential.

The Oct-3 gene is expressed in highly undifferentiated cells and is implicated in mammalian early embryogenesis. We have generated a series of hybrid cells between pluripotent embryonal carcinoma cells (Oct-3+) and fibroblasts (Oct-3-), and have studied the regulation and function of Oct-3. Upon fusion, the hybrid cells differentiated to nestin+/Brn-2+ cells resembling neuroepithelial stem cells. Expression of Oct-3 was extinguished at the transcriptional level in all the hybrid cells examined. The Oct-3 modulating activity required for the Oct-3-mediated enhancer activation was also extinguished. When the Oct-3 transactivating function was introduced into the hybrid cells, they transformed into morphologically distinct nestin-/Brn-2- cells ('revertants'). When the 'revertant' cells subsequently lost Oct-3 expression, they differentiated back to nestin+/Brn-2+ cells. The close correlation between the phenotypic changes and the gain/loss of Oct-3 function indicates that Oct-3 can induce dedifferentiation of the neural cells.

Animals↗

Multilineage response in aplastic anemia patients following long-term administration of filgrastim (recombinant human granulocyte colony stimulating factor).

The present multicenter study was undertaken to confirm whether filgrastim/recombinant human granulocyte colony stimulating factor (rhG-CSF) could mobilize residual multipotential stem cells by its G0-shortening effect in patients with aplastic anemia (AA) and induce a multilineage response. Twenty-seven patients with acquired severe or moderate AA received long-term administration (2 to 12+ months) of rhG-CSF in doses from 100 to 400 micrograms/body/day by s.c. injection or 250 to 1,500 micrograms/body/day by i.v. infusion. Twenty-six out of the 27 evaluable patients showed a substantial increase in neutrophils associated with a recovery of myeloid precursors in bone marrow within one month of therapy. Interestingly, 10 out of the 27 patients showed a dramatic improvement in severe anemia after two to ten months of therapy. Moreover, severe thrombocytopenia improved after two to four months of therapy in three out of these ten patients accompanied by a significant increase in megakaryocytes in bone marrow. Clonal cultures of bone marrow cells revealed a recovery in myeloid as well as erythroid precursors in most of these ten patients. In two patients who showed a trilineage response, mixed and megakaryocyte colony formations also recovered. These results suggest that long-term administration of rhG-CSF mobilizes myeloid, erythroid, megakaryocyte and multipotential progenitor cells and induces a multilineage response in some patients with AA.

Adolescent↗

Sequencing analysis of Ha-, Ki-, and N-ras genes in rat urinary bladder tumors induced by N-[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) and sodium saccharin.

Male F344 rats were fed N[4-(5-nitro-2-furyl)-2-thiazolyl]formamide (FANFT) for up to 4 wk, then given the basal diet with or without 5% sodium saccharin for up to 100 wk. In a previous study, we demonstrated point mutations in codons 12 and 61 of Ha-ras gene among eleven transitional cell carcinomas (TCC), one undifferentiated carcinoma, and two sarcomas of the urinary bladder (Mol Carcinogen 3:210-215, 1990). In this study, Ha-ras, Ki-ras, and N-ras sequences were examined by polymerase chain reaction (PCR) and direct DNA sequencing. The results confirm the point mutation in codon 61 (CAA to CGA in 5 TCCs and to CTA in one TCC) of the Ha-ras gene. Mutation at codon 12 was not confirmed. No mutation was found in the Ki-ras gene. Sequences of the N-ras gene exons 1 and 2 were determined, and no mutations was detected. These results suggest the involvement of activated Ha-ras gene, but not Ki-N or N-ras gene, in rat urinary bladder carcinogenesis induced by FANFT. Subsequent sodium saccharin administration did not affect the changes in Ha-ras gene.

Amino Acid Sequence↗

Murine chromosomal location of four class III POU transcription factors.

A family of mouse genes encoding class III POU transcription factors consists of four members: Oct-6, Brn-2, Brn-1, and Brn-4. In accordance with mouse nomenclature, these loci have been designated Otf-6, Otf-7, Otf-8, and Otf-9, respectively. While the four genes share a highly conserved class III POU domain, the remaining regions are considerably diverged from each other. The POU domains of these transcription factors recognize the octamer-motif sequence and possess a similar DNA-binding specificity. Chromosomal mapping has demonstrated that the four genes are unlinked: Otf-6 is located on the distal region of chromosome 4, Otf-7 on the proximal region of chromosome 4, Otf-8 on chromosome 1, and Otf-9 on the X chromosome.

Animals↗

Hereditary triosephosphate isomerase (TPI) deficiency: two severely affected brothers one with and one without neurological symptoms.

A 13-year-old Hungarian boy (B.J.Jr.) with congenital haemolytic anaemia (CHA) and hyperkinetic torsion dyskinesia was found to have severe triose-phosphate isomerase (TPI) deficiency. One of his two brothers (A.J.), a 23-year-old amateur wrestler, has CHA as well, but no neurological symptoms. Both have less than 10% TPI activity and a highly increased dihydroxyacetone phosphate (DHAP) level in their red blood cells. Their TPI had a slow electrophoretic mobility and was heat unstable. Both parents and a third brother are healthy heterozygous carriers of the defect. A.J. represents a unique phenotype from the point of view that all published "homozygotes" had severe neurological alterations from infancy or early childhood except one infant who died at 11 months, probably too young for neurological symptoms to be noted. In contrast to the two affected Hungarian brothers all but one "homozygote" has died before the age of 6 years. The striking difference in the clinical course of the defect between the two brothers with the same severe red blood cell enzyme deficiency may originate from unusual differences between two double heterozygous brothers resulting inter alia in different levels of TPI expression in various tissues. Significantly lower TPI activities were found in both the T- and B-cells of the propositus as compared to the respective cells of the neurologically symptom-free brother.

Adult↗

Molecular abnormality of G6PD Konan and G6PD Ube, the most common glucose-6-phosphate dehydrogenase variants in Japan.

G6PD Konan and G6PD Ube are the most common glucose-6-phosphate dehydrogenase (G6PD) variants found in Japan. To clarify the molecular abnormality of these two variants, the entire coding region was amplified by polymerase chain reaction from genomic DNA (G6PD Konan) or cDNA (G6PD Ube). Direct sequencing revealed that both variants have the same nucleotide substitution (241 C to T) in exon 4, which predicts an Arg to Cys substitution at amino acid 81.

Arginine↗

Intrahepatic distant metastasis and metachronous multicentric occurrence in solitary hepatocellular carcinoma of less than five centimeters in diameter.

During the 7 years from 1984 to 1990, 36 patients underwent liver resection for solitary hepatocellular carcinoma (HCC) measuring less than 5 cm in diameter, with no intrahepatic vascular invasion on imaging diagnoses and no macroscopic infiltration into the tumor capsule or surrounding tissues. Although HCC is less likely to cause intrahepatic adjacent metastasis to the cut liver surface, an analysis revealed the possibility of intrahepatic distant metastasis and metachronous multicentric occurrences, even after complete removal of the primary tumor. The 5-year cumulative survival rate was 53%, while the 5-year cumulative recurrence-free survival rate was 19%. Of the 36 patients, 18 (50%) had suffered a recurrence by April, 1992, one with extrahepatic metastasis. Recurrence of intrahepatic metastasis was multifocal in 5 patients, single and adjacent in 1, and single (or a few) and distant in 11. Multifocal recurrence was observed within 1 year after liver resection. The sole single and adjacent metastatic case occurred in one of eight patients in the recurrent group in whom distance of the surgical margin was less than 1 cm [TW(+)]. Multicentric occurrence was found in 6 of 13 patients (46%) whose recurrent tumors were examined histologically, and all belonged to the "single (or a few) and distant" type of recurrence. In this report, we also present two typical cases of metastasis, one being multifocal metastasis occurring within 3 months after liver resection and the other being intrahepatic metastasis occurring after a 4-year-dormant state, to demonstrate the complicated nature of the intrahepatic metastatic pattern.

Adult↗

Resistance of mice vaccinated with rabies virus internal structural proteins to lethal infection.

Mice were vaccinated with recombinant vaccinia virus (rVac) expressing the glycoprotein (G), nucleoprotein (N), phosphoprotein (NS) or matrix protein (M) of rabies virus and their resistance to peripheral lethal infection with street rabies virus was examined. Mice vaccinated with rVac-G or rVac-N developed strong antibody responses to the corresponding proteins and essentially all mice survived challenge infection. Mice vaccinated with rVac-NS or rVac-M developed only a slight antibody response, however, a significant protection (59%) was observed in the rVac-NS-vaccinated mice, whereas rVac-M-vaccinated mice were not protected. No anti-G antibodies were detected in the sera of mice which has been vaccinated with rVac-N or rVac-NS and survived challenge infection. Passive transfer of anti-N monoclonal antibodies (MAbs) recognizing an epitope located on amino acids 1-224 of the protein prior to challenge resulted in significant protection, although the protection was not complete even with a high amount of antibodies. In contrast, none of the mice given MAbs recognizing an epitope of amino acids 247-415 or F(ab')2 fragments from a protective MAb IgG were protected. Administration of anti-CD 8 MAb to rVac-N-vaccinated mice showed no significant effect on protection. Our observations suggest that a considerable part of the protection achieved by the vaccination with rVac-N can be ascribed to the intact anti-N antibodies recognizing an epitope located on amino acids 1-224 of the protein.

Animals↗

Proportions of fiber types in the external urethral sphincter of young nulliparous and old multiparous rabbits.

The proportions of fast and slow myosin molecules in specimen's of the external urethral sphincter (EUS) from 6 young nulliparous (6-month-old) and 6 old multiparous (2-year-old) rabbits were studied using myosin heavy chain electrophoresis. The percentages of fast and slow myosin molecules were 80.4 +/- 4.5% and 19.6 +/- 4.5% in the EUS from nulliparas, and 68.7 +/- 6.3% and 31.3 +/- 6.3% in the EUS from multiparas. The difference between the two groups was significant (P < 0.01). We suggest that a selective decrease in the volume of type 2 (fast) muscle fibers and/or conversion of type 2 to type 1 (slow) muscle fibers had taken place in the EUS of old multiparas. The proportional change in the constituent muscle fibers of the EUS with aging may play a role in human genuine stress incontinence.

Aging↗

Immunoglobulin secreting cells in lymphocytes of patients with IgA deficiency or hyper-IgA.

To assess the ability of immunoglobulin production in vivo, we enumerated the immunoglobulin secreting cells in the peripheral blood of patients with an IgA deficiency and of those with hyper-IgA-emia. All seven patients with primary IgA deficiency and two of the three patients with secondary IgA deficiency had low numbers of IgA secreting cells. In all five patients with hyper-IgA the number of IgA secreting cells was increased. Our results suggest that measurement of immunoglobulin secreting cells in PBMCs is useful in the assessment of ability of immunoglobulin production in vivo.

Antibody-Producing Cells↗

Reassessment of quantitative thallium-201 brain SPECT for miscellaneous brain tumors.

In order to reassess the value of quantitative thallium-201 brain SPECT in the differentiation of miscellaneous brain tumors, we studied a total of 89 patients--35 pre-operative patients suspected of having a brain tumor and 54 post-operative patients with a brain tumor. We came to the conclusion that quantitative Tl-201 brain SPECT was very useful in discriminating cerebral radiation necrosis from recurrent tumor, estimating residual tumor burden, and detecting tumor regrowth earlier in postoperative patients. In preoperative patients, however, Tl-201 SPECT cannot be used effectively to differentiate glioma from other intracranial tumors, although intense uptake of Tl-201 may provide evidence of glioblastoma or a hypervascular lesion.

Adolescent↗

The presence of small q waves and decreased precordial r waves indicates a small amount of fibrosis of the anterior myocardial wall.

Biopsy specimens were obtained from the anterior wall of the left ventricle during aortocoronary bypass surgery in 79 patients with critical narrowing or occlusion of the left anterior descending artery. The percent of fibrous replacement on histological analysis was calculated using the point-count method and compared with electrocardiographic findings in the precordial leads. In specimens from 19 patients with abnormal Q waves, the percent of fibrosis ranged between 38% and 100% (mean, 61 +/- 17%). Fifteen patients had small q waves or decreased r waves, and the percent of fibrosis in these patients ranged between 20% and 45% (mean, 38 +/- 10%). Specimens from 45 patients with normal QRS complexes had between 3% and 27% (mean, 11 +/- 5%) fibrosis of the entire thickness of the anterior wall. Thus, small q waves or decreased r wave amplitude in the precordial leads indicates a lesser degree of myocardial loss than the presence of abnormal Q waves.

Coronary Angiography↗

A CNS-specific POU transcription factor, Brn-2, is required for establishing mammalian neural cell lineages.

The pluripotent embryonal carcinoma cell line P19 can differentiate in vitro into neurons and astrocytes. By employing this neural cell differentiation system, we have studied the expression and function of a POU transcription factor implicated in mammalian neurogenesis. When P19 cells differentiated to the neural cells, one of the CNS-specific POU genes (Brn-2) was selectively induced. The induction of Brn-2 was specific to the neural cell lineages and took place at the early stage of differentiation. When the Brn-2 induction was blocked/delayed by antisense RNA, such cells were unable to differentiate to neurons and astrocytes. Instead they differentiated to nonneural cells, including smooth and skeletal muscle cells. Furthermore, with reduced levels of antisense RNA, differentiation to neural cells occurred. These results indicate that Brn-2 is essential for the neural cell differentiation of P19 cells and suggest that Brn-2 is one of the genes required for establishing neural cell lineages in mammals.

Animals↗

Endocrine aspects of exocrine cancer of the pancreas. Their patterns and suggested biologic significance.

Nine human exocrine pancreatic adenocarcinomas were examined by serial sectioning and double- and triple-labeled immunohistochemical techniques with antibodies against chromogranin A, insulin, islet amyloid polypeptide, glucagon, somatostatin, pancreatic polypeptide, serotonin, pancreastatin, and neuron-specific enolase. The results were correlated with the stage of the disease, histologic characteristics of the tumors, and survival of the patients. Cells immunoreactive with most or all of the antibodies were found in all nine cases. Abnormal co-location of some hormones in the same cell and the lack of normal co-location of other hormones were found. Endocrine cells also were identified in the invasive regions of the cancer, including perineural spaces. Abnormality in the production and release of the peptide was indicated not only in the endocrine cells of exocrine cancer, but also in the islets near the cancer. Patients whose cancer contained many endocrine cells seemed to survive longer than those with tumors containing fewer endocrine cells. The overall data suggested that the observed abnormalities may contribute to the impaired glucose tolerance found in six of these patients.

Adenocarcinoma↗

Production of scatter factor-like activity by a nitrosamine-induced pancreatic cancer cell line.

Two hamster pancreatic cancer cell lines, PC-1 and PC1.0, established from N-nitrosobis(2-oxopropyl)amine-induced pancreatic ductal/ductular adenocarcinomas exhibit different growth patterns. PC-1 cells, which produce well differentiated adenocarcinomas in vitro after allogeneic inoculation, form cell aggregates and characteristic island-like structures in vitro. PC1.0 cells, which produce poorly differentiated tumors in vivo, form dispersed colonies in vitro. Conditioned medium prepared from PC1.0 cells inhibits PC-1 cells from forming island-like colonies. The conditioned medium also prevents several human pancreatic carcinoma cell lines, HPAF, CD11 and CD18, from forming compact colonies. These properties are similar to those described previously as scatter factors. The scatter factor-like activity is heat-labile, acid-stable, non-dialyzable, trypsin sensitive and unaffected by reducing agents. The activity is not suppressed by addition of heparin, and it does not bind to heparin. In addition, the scatter phenomenon is not reproduced by acidic or basic fibroblast growth factor, epidermal growth factor or transforming growth factor-beta 1. Based on these findings, it appears that the scattering activity produced by PC1.0 cells differs from the scatter factors that have been identified in other systems.

Adenocarcinoma↗