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Biomedical subjects

T Fukasawa

Publications and source records attributed to T Fukasawa.

At least 73 records · Page 4Linked to original sources
PubMed1988

[Postoperative sequential testing of cytotoxic activity against the autologous and allogeneic tumor cells in patients with primary lung resected cancer].

The objective of this study of 27 primary lung resected cancer patients was to evaluate sequentially the cytotoxic activity against autologous and allogeneic tumor cells during the postoperative period. In patients given a complete resection, the cytotoxic activity against the autologous tumor cells during the postoperative 1-2 weeks was significantly higher than during the postoperative 4-5 weeks (P less than 0.05, Student t test). In patients given an incomplete resection, however, no significant difference was demonstrated between the postoperative 1-2 weeks and during the postoperative 4-5 weeks. In contrast, the cytotoxic activity against the PC-3 cells or K562 cells during the postoperative 4-5 weeks tended to rise in comparison to those at the postoperative 1-2 weeks all patients in any cancer stage and whatever the amount of their lung resection. The phenotype of the effector cells which mediated the cytotoxic activity to the autologous tumor cells was the monoclonal antibody OKT 3(+), 8(+), 4(-) OKM 1(-) and the cells which were cytotoxic to the K562 cells had an OKT 3(-), 4(-), 8(-), OKM 1(+) phenotype. These results suggest that the effectors which mediate cytotoxic activity to the autologous tumor cells and to the K562 cells seems to be different and a concomitant immunity may exist in primary lung resected cancer patients.

Adenocarcinoma↗
PubMed1987

Sequence analysis of elastin cDNA from chick aorta and tissue-specific transcription of the elastin gene in developing chick embryo.

A portion of elastin cDNA with a size of 1.5 kilobase pairs (kb) was cloned from chick aorta. Sequence analysis revealed that the cDNA consists of 0.9 kb of coding region and 0.6 kb of 3'-untranslatable region. The primary structure of the peptide deduced from the coding sequence exhibited a strong homology with the published data from sheep and bovine elastin cDNA. The abundance of elastin mRNA in the aorta and skin was studied in developing chick embryo by Northern analysis using the cDNA as a probe. The elastin mRNA level in the aorta gradually decreased in the late half of development, while the elastin mRNA level in the skin was dramatically elevated between the 18th and 21st days. These results strongly suggest that the transcription of the elastin gene was controlled specifically in the respective organ during development.

Amino Acid Sequence↗
PubMed1987

Autogenous regulation of the Saccharomyces cerevisiae regulatory gene GAL80.

We have suggested previously from Northern blot analysis that transcription of the negative regulatory gene GAL80 was controlled positively by another regulatory gene GAL4, and negatively by GAL80 itself, in similar way to GAL1, GAL7 and GAL10 genes encoding galactose-metabolizing enzymes in Saccharomyces cerevisiae. To study further the controlled expression of GAL80, we have exploited the gene fusion technique. We constructed gene fusions consisting of 5' fragments of GAL80 and a 5' truncated lacZ of Escherichia coli, and introduced the GAL80'-'lacZ fusions into wild-type yeast or various GAL4 or GAL80 mutants using multiple-copy or single-copy plasmid vectors. We then studied beta-galactosidase activity in the resultant transformants under uninduced, induced or glucose-repressed conditions. Expression of the GAL80'-'lacZ fusions was clearly under the control of Gal4/Gal80. Next we constructed GAL7'-'lacZ fusions, whose upstream activating sequence (UAS) from GAL7 was replaced with a GAL80 fragment containing a UAS-like sequence located in the 5' flanking region of GAL80. Synthesis of beta-galactosidase directed by the hybrid genes was inducible by galactose exactly like the original GAL7'-'lacZ fusion with a UAS from GAL7. Finally we constructed a GAL7-GAL80 hybrid gene, in which the entire 5' flanking region was derived from GAL7. When the chromosomal GAL80 gene in wild-type yeast was replaced with the hybrid gene, the uninduced level, but not the induced level, of the GAL10-encoded enzyme (uridine diphosphoglucose-4-epimerase) was significantly increased.

Galactose↗
PubMed1987

Expression of the human adenovirus E1a product in yeast.

We synthesized the 13S mRNA-encoded protein of the early region 1a (E1a) of human adenovirus in Saccharomyces cerevisiae under the control of the yeast GAL7 gene promoter. Similar to the case in HeLa cells, the E1a protein in yeast was phosphorylated and formed multiple bands on sodium dodecylsulfate-polyacrylamide gel electrophoresis. These bands migrated more slowly than expected from the Mr calculated on the basis of the nucleotide sequence of the gene. Synthesis of the E1a protein caused induction of a specific family of heat-shock proteins (Hsp70), which, however, did not confer heat resistance to the yeast. In addition, the E1a production resulted in an elongation of the generation time of yeast from 2.4 h to 3.9 h, which was attributed specifically to elongation of the G1 interval in the cell cycle. In the light of these findings, we suggest that the E1a protein synthesized in yeast exerts a specific function.

Adenovirus Early Proteins↗
PubMed1986

Intratumoral ethanol injection for malignant tracheobronchial lesions: a new bronchofiberscopic procedure.

We performed intratumoral ethanol injection via a flexible bronchofiberscope in 13 patients with malignant tracheobronchial lesions in order to evaluate its effects on airway dilatation and hemostasis. The results obtained are described below. Immediately after intratumoral injection of ethanol, bronchofiberscopic findings revealed that the tumor turned faintly white, there was a little regression of tumor, and a promising effect was demonstrated on patients with bleeding from tumors. The injected tumor turned necrotic within several days, and histological examination revealed no viable tumor cells in necrotic tissues. The histological anti-tumor effect of ethanol was also demonstrated in experiments with nude mice. This endoscopic treatment was very effective in polypoid tumor protruding into the tracheobronchial lumen, but ineffective in the case of compressed stenosis or obstruction. In conclusion, intratumoral injection of ethanol is considered to be a promising endoscopic treatment for malignant tracheobronchial lesions.

Adult↗
PubMed1986

Duplicate upstream activating sequences in the promoter region of the Saccharomyces cerevisiae GAL7 gene.

We constructed a series of deletions in the 5' noncoding region of the Saccharomyces cerevisiae GAL7 gene, fused them to the Escherichia coli gene lacZ, and introduced them into yeasts by using a multicopy vector. We then studied the effect of the deletions on beta-galactosidase synthesis directed by the gene fusions in media with various carbon sources. This analysis identified a TATA box and two upstream activating sequences as necessary elements for galactose-controlled GAL7 transcription. Two upstream activating sequences exhibiting 71% homology with each other were located 255 and 168 base pairs, respectively, upstream of the GAL7 transcription start point. Each sequence consists of 21 base pairs, displaying an approximate rotational symmetry with a core consensus sequence of GAA--AGCTGCTTC--CGCG. At least one of the two sequences is required for galactose induction and also for glucose repression of the GAL7'-lac'Z gene. Analysis with host regulatory mutants delta gal14 and delta gal180 suggests that these sequences are the site at which the GAL4 product exerts its action to activate the GAL7 gene. We also observed that a deletion lacking both upstream activation sequences allowed the gene fusion to be expressed in the absence of galactose at about 10% of the fully induced level of the intact fusion. This constitutive expression depended on the presence of the TATA box of GAL7 in cis but not on a functional GAL4 gene. The level of the uncontrolled expression was decreased by increasing the distance between the TATA box and the pBR322 sequence in the vector plasmid.

Base Sequence↗
PubMed1986

[Clinical evaluation of serum NSE and CEA in primary lung cancer patients].

The clinical value of serum neuron specific enolase (NSE) as a tumor marker was evaluated in comparison with simultaneously measured serum CEA, using 74 cases of lung cancers, 13 cases of non-tumorous and non-neurological disease and 28 normal volunteers. The results obtained were as follows: Thirty-five out of the 74 lung cancer cases showed positive serum NSE levels. However, none of the cases of non-tumorous, non-neurological disease and normal volunteers demonstrated more than 10ng/ml NSE which was significantly different from the results of lung cancer cases. We found a higher degree of NSE positiveness in cases of small cell carcinoma than in cases of other histological types. However, no obvious difference of CEA positiveness between any histological types was revealed. Both serum NSE and CEA showed an increase in positive rate in parallel with the progress of stages, including 80.4% of cases of stages III and IV disease which had positive serum levels of both or either of the two markers. However, we could not find any obvious correlation between serum NSE and CEA. The serum levels of both markers clearly decreased after surgery, serum CEA level being considered to be a better reflection of host tumor burden than NSE. In conclusion, the simultaneous testing of serum NSE and CEA is considered to be very useful for evaluating the clinical course or the effect of cancer therapy in lung cancer patients, with higher specificity and sensitivity.

Adenocarcinoma↗
PubMed1985

Primary structure of the Saccharomyces cerevisiae GAL7 gene.

We present the nucleotide sequence of a 1599-base pair (bp) DNA fragment containing the entire GAL7 gene that encodes galactose-1-phosphate uridyltransferase of Saccharomyces cerevisiae. The deduced peptide was composed of 364 amino acid residues. The expected molecular weight was 42,005 daltons, which agreed with the observed value for the purified enzyme. The 3'-end of the GAL7 transcript mapped at a position 82 bp downstream from the UAA termination codon by the S1 nuclease protection experiment. We constructed a GAL7'-lac'Z fusion on various types of yeast plasmid vectors. The fused gene on any type of vector was induced by galactose and repressed by glucose as for the GAL7 gene on the chromosome. The response of GAL7'-lac'Z fusion to gal4 delta and gal80 delta regulatory mutations was also similar to the response of the chromosomal GAL7 gene. By using various deletions in the 5'-flanking region of the gene fusion, we delimited the sequence essential for galactose controlled expression with a 180 bp-fragment of DNA lying 92 bp upstream of the transcription initiation site.

Amino Acid Sequence↗
PubMed1985

Transcription of the human adenovirus E1a gene in Saccharomyces cerevisiae.

The early region 1a (E1a) and its flanking sequences of human adenovirus type 5 (Ad5) have been cloned in the yeast-Escherichia coli shuttle vector YEp13 and transferred into the yeast Saccharomyces cerevisiae. The E1a-specific RNAs were produced in the transformed yeast cells. The 5' ends of these transcripts were capped but were lacking 10 to 45 nucleotides from the 5' end of the proper E1a mRNA. These transcripts terminated approx. 1000 nucleotides downstream from the proper 3' end. No splicing of the E1a-specific RNA could be detected in the yeast cells.

Adenoviruses, Human↗
PubMed1985

Controlled transcription of the yeast regulatory gene GAL80.

Transcription of the tightly linked genes GAL7-GAL10-GAL1 encoding three galactose-metabolizing enzymes in Saccharomyces cerevisiae is regulated by an interplay of the positive regulatory gene GAL4 and the negative regulatory gene GAL80. The concentration of GAL80 mRNA (determined by a quantitative blot hybridization) was nearly comparable to that of URA3 mRNA, in the wild-type yeast grown in medium with glucose or glycerol. The intracellular concentration of GAL80 mRNA increased by changing the carbon source to galactose by a factor of more than 5, in contrast to a GAL4 mRNA concentration which is essentially unaffected by galactose [Laughon and Gesteland, Proc. Natl. Acad. Sci. USA 79 (1982) 6827-6831]. The inducible expression of the GAL80 gene was suggested to involve its own product and also the product of GAL4 by using various mutations in those genes.

DNA, Fungal↗
PubMed1984

Nucleotide sequence of the yeast regulatory gene GAL80.

The GAL80 gene in Saccharomyces cerevisiae encodes a negative regulatory protein for the set of inducible genes involving metabolism of galactose and melibiose. We have determined the nucleotide sequence of GAL80 and its flanking regions and assigned the 5' end of its mRNA to the sequence. The deduced coding sequence for GAL80 protein contains 1305 nucleotides and the calculated molecular weight of the peptide chain is 48309. The 5' end of the GAL80 mRNA maps about 67 nucleotides upstream from the translation initiating ATG. We have also determined the nucleotide sequence of uninducible alleles GAL80S-0, GAL80S-1 and GAL80S-2, and found single base substitution in each of these mutant genes which would lead to alteration of amino acid in GAL80 protein.

Alleles↗
PubMed1984

Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. II. The isolation and dosage effect of the regulatory gene GAL80.

The galactose analogue 2-deoxygalactose was found to inhibit the growth of a mutant strain of Saccharomyces cerevisiae constitutively producing the set of galactose utilization enzymes. Based on this fact, the yeast GAL80 gene negatively regulating the expression of the genes encoding those enzymes was isolated for its ability to confer 2-deoxygalactose resistance on a strain carrying a recessive mutation in that gene. The GAL80 gene was located within a 3.0 kb fragment in the cloned DNA. When the isolated gene was incorporated into a multi-copy plasmid, the induced level of three enzymes encoded by the gene cluster GAL7-GAL10-GAL1 in the host chromosome was lowered. Such a gene dosage effect of GAL80 was further pronounced if sucrose, a sugar causing catabolite repression, was added to the growth medium. The ratio of the enzyme activity of the yeast bearing multiple copies of GAL80 to that of the yeast bearing its single copy significantly varied with the enzyme. From these results we suggest that the intracellular inducer interacts with the GAL80 product and that GAL80 molecules directly bind the GAL cluster genes with an affinity different from one gene to another.

Base Sequence↗
PubMed1983

Nucleotide sequence of the transcriptional initiation region of the yeast GAL7 gene.

The GAL7 gene of Saccharomyces cerevisiae encodes Gal-1-P uridylyl transferase, the second enzyme of Leloir pathway for the galactose catabolism. We have determined the sequence of 1003 base pairs surrounding and upstream of the transcriptional initiation site of the GAL7 gene. The region sequenced also encompasses the 3' end of GAL10 gene. The 5' end of GAL7 mRNA was determined on the DNA sequence by the S1 nuclease- and exonuclease VII mapping, which is located 21 to 22 base pairs upstream from the translation initiating ATG codon. The primary structure of the GAL7 5' flanking region has many features common to those of multicellular eukaryotic genes. The 3' end of GAL10 mRNA was also determined by the mapping technique with the single-strand specific nucleases to be about 600 base pairs upstream from the 5' end of GAL7 mRNA.

Base Composition↗
PubMed1983

Regulation of expression of the galactose gene cluster in Saccharomyces cerevisiae. Isolation and characterization of the regulatory gene GAL4.

The GAL4 gene positively regulating the expression of the gene cluster GAL7-GAL10-GAL1 in the yeast Saccharomyces cerevisiae was isolated for its ability to suppress a recessive mutation in that gene. When the isolated gene was incorporated into a multi-copy plasmid, the GAL cluster genes in the host chromosome partially escaped the normal control; a yeast that harbors the plasmid bearing the GAL4 gene synthesized the galactose-metabolizing enzymes encoded by the GAL cluster genes at a low but significant level in the absence of galactose. If the GAL7 gene was amplified along with GAL4 on the multi-copy plasmid, the constitutive synthesis of Gal-1-P uridylyl transferase encoded by GAL7 was further pronounced and the enzyme activity reached the level of the fully induced wild-type yeast. Such an escape synthesis of the GAL enzymes was not detected if GAL4 or both GAL4 and GAL7 were carried by a single-copy plasmid. The results suggest that the escape synthesis of GAL enzymes observed in the GAL4-amplified yeast was a consequence of overproduction of the GAL4 protein. The GAL80 gene negatively regulating the GAL cluster genes was also isolated, and when amplified together with GAL4, no escape synthesis of the GAL enzymes was observed, suggesting that the balanced synthesis of two regulatory proteins was essential to maintain the repressed state of the GAL cluster genes.

DNA, Fungal↗