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Biomedical subjects

T Morinaga

Publications and source records attributed to T Morinaga.

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

A fibroblast-derived tumor cytotoxic factor/F-TCF (hepatocyte growth factor/HGF) has multiple functions in vitro.

We previously demonstrated that a tumor cytotoxic factor(F-TCF) purified from the culture broth of human embryonic lung diploid fibroblast, IMR-90 cells was one of the human hepatocyte growth factors (hHGFs). In the present report, we demonstrate its biological functions. F-TCF showed moderate cytotoxicity on human tumor cell lines KB, BG-1, MCF-7 and Hs913 T, and strong cytotoxicity on mouse tumor cell lines Sarcoma 180, Meth A sarcoma and P388. On the contrary, F-TCF was a potent mitogen not only for adult rat hepatocytes, but also for human endothelial cells, HUVEC and human melanocytes. Moreover, F-TCF induced the differentiation of premyelocyte leukemia, HL-60 cells into morphologically granulocyte-like cells. These biological functions suggest that F-TCF is an effector molecule responsible for inflammation and repair in injured tissues including tumor and liver.

Animals↗
PubMed1991

A human alpha-fetoprotein enhancer-binding protein, ATBF1, contains four homeodomains and seventeen zinc fingers.

We have isolated a full-length cDNA encoding a protein (ATBF1) that binds to an AT-rich motif in the human alpha-fetoprotein gene enhancer. The amino acid sequence deduced from the cDNA revealed that this is the largest DNA-binding protein (306 kDa) known to date, containing four homeodomains, 17 zinc finger motifs, and a number of segments potentially involved in transcriptional regulation. Although the exact function of this protein has not been determined, these structural features suggest that ATBF1 plays a transcriptional regulatory role.

Amino Acid Sequence↗
PubMed1991

Purification and characterization of tissue plasminogen activator secreted by human embryonic lung diploid fibroblasts, IMR-90 cells.

The anti-urokinase-IgG-resistant plasminogen activator secreted by human embryonic lung diploid fibroblasts, IMR-90 cells (ATCC, CCL186) was purified to homogeneity from serum-free conditioned medium by a four-step procedure. The fibroblast plasminogen activator was identified as tissue plasminogen activator (t-PA) by the N-terminal sequence of the purified material and the complete amino acid sequence deduced from its complementary DNA (cDNA). The apparent molecular weight was the range of 64,000 to 68,000 by SDS-PAGE and was in the range of 69,000 to 72,000 by gel filtration. The fibroblast t-PA showed a stricter substrate specificity than urokinase in enzymatic hydrolysis of various chromogenic substrates. Compared to urokinase, the fibrobrast t-PA was more stable by heating at 95 degrees C for five min and was stable from pH 5 to 10. The fibrorast t-PA had a higher affinity for fibrin than urokinase.

Amino Acid Sequence↗
PubMed1991

Effect of interferon administration on serum hepatitis C virus RNA in patients with chronic hepatitis C.

Hepatitis C virus RNA as detected by reverse transcription and nested polymerase chain reaction was monitored in 16 patients with chronic hepatitis C treated with interferon. Hepatitis C virus RNA became undetectable after 4 to 8 wk of interferon administration in 13 of the 16 patients. During 6 mo of follow-up, 5 of the 13 patients who became negative for hepatitis C virus RNA after interferon administration remained negative, and all five continued to have normal ALT levels. Repeat liver biopsy in these five patients revealed histological improvement. Antibody to hepatitis C virus, which was initially positive in all treated patients, fell to undetectable levels in three of the five patients. In contrast, aminotransferase levels rose again in all eight patients who had become hepatitis C virus RNA negative but had again exhibited hepatitis C virus RNA after completion of therapy. In 16 untreated patients, hepatitis C virus RNA remained detectable. These results indicate that detection of hepatitis C virus RNA may be useful as a marker of viral replication in chronic hepatitis C; they also suggest that interferon should again be administered to patients who become hepatitis C virus RNA negative on treatment but again exhibit this marker of viral replication when treatment is stopped.

Adult↗
PubMed1991

[Clinical features and viral markers in patients with chronic hepatitis positive for both HBs antigen and anti-C-100 antibody].

Among 612 patients with chronic hepatitis type B, 24 patients tested positive for anti-C-100 antibody. Clinical features and viral markers of these 24 patients were studied. Anti-C-100 antibody was detected more often in patients negative for HBe antigen than those positive for this antigen (22/385 vs. 2/227 [P less than 0.01]). HCV RNA, as detected by reverse transcription and nested polymerase chain reaction, was positive in 20 of 22 patients negative for HBe antigen whereas it was detected in only one of two patients positive for HBe antigen. In a patient who seroconverted from HBe antigen to anti-HBe HCV RNA was undetectable in the HBe antigen positive phase and had become detectable after seroconversion. These data indicate that both hepatitis virus (type B and type C) can co-infect in a patient and that type C becomes active after the seroconversion from HBe antigen to anti-HBe.

Adult↗
PubMed1990

Identity of a tumor cytotoxic factor from human fibroblasts and hepatocyte growth factor.

Human embryonic lung diploid fibroblast, IMR-90 cells secreted a tumor cytotoxic factor. The fibroblast-derived tumor cytotoxic factor (F-TCF) has a cytotoxic activity to Sarcoma 180 and a cytostatic and degenerative activities to KB cells. F-TCF has been purified about 540,000-fold with 23.3% recovery from 75 liters of the conditioned medium containing 5% newborn calf serum. The purified F-TCF is a basic glycoprotein with isoelectric point values of 7.4 to 8.6. It was stable in the pH range from 6.0 to 9.0 and was stable at the heating temperature of 60 degrees C for 10 min, but completely inactivated by reducing it with 2-mercaptoethanol. F-TCF has molecular weight of 76 to 80 kD on SDS-PAGE under non-reducing conditions and is a heterodimer consisting of a large alpha subunit with 52 to 56 kD and a small beta subunit with 30 to 34 kD. F-TCF was identified as one of human hepatocyte growth factors by the physicochemical properties including N terminal and a few internal amino acid sequences. We have confirmed that F-TCF has an ability to dramatically stimulate DNA synthesis in adult rat hepatocytes in the low dose range of 1 to 10 ng/ml.

Amino Acid Sequence↗
PubMed1990

Expression of human alpha-fetoprotein in yeast.

Human alpha-fetoprotein (AFP) was expressed in Saccharomyces cerevisiae, with a plasmid containing the cDNA sequence for human AFP fused with the rat AFP signal peptide. The recombinant AFP was purified from the yeast lysate by DEAE-cellulose and immunoaffinity chromatography. The amino acid composition and the molecular weight of the recombinant AFP were similar to those of hepatoma AFP. N-terminal amino acids sequence analysis indicated that the signal peptide had been processed. The recombinant and hepatoma AFP reacted identically in Ouchterlony immunodiffusion and radioimmunoassay tests. These observations indicated that the yeast recombinant protein had the properties of native AFP.

Amino Acid Sequence↗
PubMed1990

Physical mapping and molecular cloning of mung bean yellow mosaic virus DNA.

Viral single-stranded DNA of mung bean yellow mosaic virus (MYMV) was converted to the double-stranded state in vitro, and physical mapping was carried out. The genome of MYMV was found to consist of two major components (designated as DNA 1 and DNA 2). In addition, some minor components were detected. Molecular cloning of the major components was carried out, using in vitro double-stranded DNA and replicative intermediate DNAs. DNA 1 is about 2.72 and DNA 2 about 2.67 kilobase pairs. No similarities were observed when the two restriction maps of DNA 1 and 2 were compared.

Cloning, Molecular↗
PubMed1990

[Reproductive and developmental toxicity study of mofezolac (N-22) (2)--Study by oral administration of N-22 during the period of fetal organogenesis in rats].

The teratogenicity of mofezolac (N-22), a new developed analgesic and anti-inflammatory agent, was investigated in rats. N-22 was given orally to pregnant rats of the Jcl: Wistar strain (30 rats per group) at dose levels of 10, 50, 100 and 150 mg/kg/day from days 7 to 17 of gestation. Caesarean sections were performed on 20 dams per group on day 20 of gestation and their fetuses were examined for external, visceral and skeletal abnormalities. The remaining 10 dams per group were allowed to deliver and their offspring were examined for growth and reproductive performance. Results were as follows. 1. Effects on F0 generation At 150 mg/kg, eleven out of the 30 dams exhibited decreased motor activity, pale eyes, unkempt fur, urine-smeared lower abdomen, weakness and emaciation. At autopsy, twelve dams revealed gastrointestinal ulcers, peritonitic lesions, hypertrophy of the spleen, adrenal and mesenteric lymph node, atrophy of the submaxillary gland, thymus and liver and discoloration of the liver and kidney. Death, sacrificing in extremis, premature or delayed delivery and poor nursing occurred in one to two dams each. Food consumption was significantly decreased and body weight gain was significantly retarded in this dose level group. At 100 mg/kg, urine-smeared lower abdomen, hypertrophy of the spleen and poor nursing were observed in one dam each. 2. Effects on F1 generation At 150 mg/kg, significantly decreased fetal weight, increased number of immature fetuses and significantly retarded ossification of the 5th and 6th sternebrae and coccygeal vertebrae as well as significantly depressed body weight gain of female offspring were observed. No abnormalities were observed in each treated group in terms of development, behavior, learning ability and reproductive performance of offspring. 3. Effects on F2 generation No abnormalities were observed in fetuses and newborn young in each treated group. Based on these results, the maximum non-effective doses of N-22 in this study were considered to be 50 mg/kg/day for dams and offspring and 100 mg/kg/day for fetuses.

Abnormalities, Drug-Induced↗
PubMed1989

Nucleotide sequence of SN-07 chromophore binding site.

DNase I footprinting was used to investigate binding sites for SN-07 chromophore on DNA fragments prepared from pBR322. Six sites were protected on about 150 base pair DNA fragments by SN-07 chromophore, but not by related anthracycline antibiotics from DNase I digestion. All the protected sites contained the dinucleotide sequence 5'-GC-3', but no other regularities could be discerned. A drug-induced conformational change of DNA was suggested by enhancement of DNase I sensitivity between the protected sites. These results support covalent interaction of the carbinolamine function of SN-07 chromophore to 2-amino group of guanine residues.

Anthracyclines↗
PubMed1989

[Clinical study of large doses of vecuronium; duration of initial and additional doses].

Neuromuscular blockade by large doses of vecuronium was investigated clinically and the duration of action of initial doses (0.2 mg.kg-1 and 0.3 mg.kg-1) and additional doses (0.01 mg.kg-1 and 0.02 mg.kg-1) were measured under enflurane-nitrous oxide anesthesia using a neuromuscular transmission monitoring system (Accelograph). In group A (initial dose = 0.2 mg.kg-1, additional dose = 0.02 mg.kg-1), the time of spontaneous recovery to 25% of control twitch height (T25) and 50% of control (T50) were 63.1 and 82.0 minutes respectively. The T50 interval of the two adjacent added doses (given at the time point of 50% recovery from the previous dose) was 31.1 minutes. Reversal time from TOF ratio = 25% to 75% after administration of edrophonium was 6.1 minutes. In group B (initial = 0.3 mg.kg-1, add. = 0.01 mg.kg-1), T25, T50, T50 interval and reversal time were 122.6, 159.4, 39.2 and 4.6 minutes respectively. In group C (initial = 0.3 mg.kg-1, add. = 0.02 mg.kg-1), above values were 119.2, 145.8, 48.4 and 6.7 minutes respectively. In this study there was no obvious side effect associated with administration of large doses of vecuronium. These methods will be very useful for long surgical procedures.

Female↗
PubMed1988

Potential gene products of bean golden mosaic virus have higher sequence homologies to those of tomato golden mosaic virus than to those of cassava latent virus.

Comparison of the nucleotide sequences of the DNAs of bean golden mosaic virus (BGMV), tomato golden mosaic virus (TGMV) and cassava latent virus (CLV) revealed a fairly close relationship between BGMV DNA1, TGMV DNA1, and CLV DNA1 and a comparatively distant relationship between BGMV DNA2, TGMV DNA2, and CLV DNA2. The 200-base region common to the two DNAs of each virus had little sequence homology, except for a highly conserved 33-36 base sequence potentially capable of forming a stable hairpin structure. All the potential coding regions in the BGMV DNAs had counterparts in the TGMV and CLV DNAs suggesting an overall similarity in genome organization but two potential coding regions in the BGMV DNAs had no counterparts in the TGMV DNAs. The most highly conserved ORFs, BGMV 1R1, TGMV 1R1, and CLV 1R1, are the putative genes for the coat proteins of BGMV, TGMV, and CLV. BGMV 1R1 has 91.9% and 71.6% homology with respect to TGMV and CLV. The ORFs (BGMV 1L1; CLV 1L1; TGMV 1L1) and the two smaller overlapping ORFs (BGMV 1L2, 1L3; TGMV 1L2, 1L3; CLV 1L5, 1L3) are conserved in the three viruses. BGMV 2R1 and BGMV 2L1 have higher homology with respect to TGMV but not with respect to 2R1 and 2L1 in CLV. From this study we conclude that BGMV is more closely related to TGMV than CLV.

Amino Acid Sequence↗
PubMed1988

Interaction of a hepatoma-specific nuclear factor with transcription-regulatory sequences of the human alpha-fetoprotein and albumin genes.

DNase I footprinting and gel mobility shift analysis showed that an HuH-7 hepatoma nuclear protein, termed AFP1, binds specifically to an AT-rich sequence, TGATTAATAATTACA, in domain B of the human alpha-fetoprotein enhancer. No such binding activity was found in HeLa cell nuclei. Transient transfection studies showed that a 54-base-pair region corresponding to the AFP1-binding site could stimulate the simian virus 40 early promoter to express a linked chloramphenicol acetyltransferase gene in an orientation-independent and cell-specific manner. The correlation between the binding of AFP1 and the stimulation of chloramphenicol acetyltransferase gene expression strongly suggests that specific interaction of AFP1 with the AT motif is important for cell-specific transcriptional enhancement. Competition gel mobility shift analysis revealed that similar AT-rich sequences with high affinities to AFP1 were also present in the promoters of the alpha-fetoprotein and albumin genes. These results suggest that AFP1 may function as a common regulatory factor in the transcription of the alpha-fetoprotein and albumin genes.

Animals↗
PubMed1987

Total nucleotide sequences of the infectious cloned DNAs of bean golden mosaic virus.

Complete nucleotide sequences of the infectious cloned DNA components (DNA A and B) of bean bolden mosaic virus were determined. The DNA A (2585 nucleotides) and DNA B (2647 nucleotides) have little sequence homology with each other, but both A and B contain a common region of 205 nucleotides. A possible large hairpin structure is detected in the common region. Nucleotide sequences of DNAs A and B revealed the presence of 8 potential coding regions for proteins (m.w. greater than 10,000). Among them, four open reading frames (ORFs 1-4) encode proteins of m.w. 30,000 or greater, and are individually coded in virion DNA A and B senses (+) and their complementary senses (-), respectively. The other four ORFs 5-8 are in virion DNA B(+) and its complementary sense (-). All of the ORFs 1-4 have regulatory signals for RNA synthesis (TATAA/T) in the region 5' upstream from a potential start codon ATG.

Base Sequence↗
PubMed1986

Stimulation of macrophage by polyanions and its conjugated proteins and effect on cell membrane.

Lipophilic anionic copolymer (styrene-maleic acid; SMA) conjugates of albumin and antitumor protein neocarzinostatin (NCS) (smancs) were found to stimulate the release of H2O2 and O-2 from the peritoneal macrophages obtained from mice which had been pretreated with the heat-killed preparation of Streptococcus pyogenes (OK-432) in vivo. Some alkyl esters of SMA exhibited effects similar to protein-polymer conjugates. Among them, butyl-SMA was the most effective followed by ethyl-SMA, whereas hydrolyzed SMA showed no effect. This activity was dose-dependent but exhibited a bell-shape profile. These results suggest that the aliphatic ester residue in SMA as well as the main chain of the copolymer may be important for the activation of macrophages. A strong antitumor effect of smancs reported elsewhere may be attributed partly to the activation of macrophages in addition to the direct damage to the cellular DNA by the NCS component. A preliminary investigation of the subcellular mechanism of macrophage activation was carried out in view of membrane fluidity by the fluorescence polarization method. The results showed that the apparent decrease in the cell membrane fluidity and the degree of macrophage activation paralleled the same dose range and at similar time courses. This indicated the interaction of SMA component and macrophage cell membrane.

Animals↗
PubMed1985

The human alpha-fetoprotein gene. Sequence organization and the 5' flanking region.

The human alpha-fetoprotein (AFP) gene was isolated into three overlapping clones in bacteriophage lambda vectors and its sequence organization analyzed by restriction endonuclease mapping and nucleotide sequencing. The human AFP gene is about 20 kilobase pairs long and contains 15 exons and 14 introns. The overall organization of the human AFP gene is similar to that of the mouse AFP gene, with all but two exons showing identical sizes. Nucleotide sequences at all exon/intron junctions display similarity to the consensus boundary sequence (Breathnach, R., and Chambon, P. (1981) Annu. Rev. Biochem. 50, 349-383), with the GT-AG rule applied to the splicing point. The cap site maps 44 nucleotides upstream from the translation initiation site. The "TATA box" is located 27 nucleotides upstream from the putative cap site and is flanked by sequences with dyad symmetry. The TATA box can thus be placed in the loop portion of a possible stem-loop structure formed by intrastrand base-pairing. Other characteristic nucleotide sequences in the 5' flanking region include a CCAAC pentamer, a 14-base pair (bp) enhancer-like sequence, and a 9-bp sequence homologous to the glucocorticoid responsive element. A long (90 bp) direct repeat and several alternating purine/pyrimidine sequences are also present in the 5' flanking region. A 736-bp sequence of the 5' flanking region adjacent to the cap site of the human AFP gene shows a 61% similarity with the corresponding region of the mouse AFP gene. There are two Alu family sequences and two poly(dT-dG) repeats in the human AFP gene that show different distribution patterns from those in the mouse AFP gene.

Amino Acid Sequence↗