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Biomedical subjects

I Ishida

Publications and source records attributed to I Ishida.

At least 73 records · Page 4Linked to original sources
PubMed1987

Molecular cloning and nucleotide sequence of cDNA encoding hydroxyindole O-methyltransferase of bovine pineal glands.

Messenger RNA for hydroxyindole O-methyltransferase (EC 2.1.1.4) was partially purified from poly(A)+ RNA isolated from bovine pineal glands by sucrose density gradient centrifugation. The enriched mRNA was used to prepare a cDNA library by use of expression vector lambda gt11. The library was screened with monoclonal antibodies to the enzyme, and three cDNA clones were isolated. These cloned cDNAs cross-hybridized with one another, and their fusion proteins reacted to the monoclonal antibodies with different binding properties. Hydroxyindole O-methyltransferase enzymatic activity was demonstrated in the bacteria lysate infected with lambda HIOMT-A16, the clone that contained the longest insert. An almost full-length cDNA clone was isolated from lambda gt10 cDNA library by use of the lambda HIOMT-A16 cDNA as a probe. The primary structure of hydroxyindole O-methyltransferase was determined by analyzing the nucleotide sequence of the cDNAs. It consisted of 1939 nucleotides including a 1050-nucleotide region coding for 350 amino acids. RNA transfer blot analysis indicated that mRNA encoding hydroxyindole O-methyltransferase was present only in the pineal gland and not in the brain, retina, and liver of cow.

Acetylserotonin O-Methyltransferase↗
PubMed1987

Expression and characterization of hydroxyindole O-methyltransferase from a cloned cDNA in Chinese hamster ovary cells.

We have used two kinds of expression systems to test whether the cloned cDNA encoding hydroxyindole O-methyltransferase (HIOMT) of the bovine pineal gland was functional or not. First, when mRNA was synthesized in vitro by the SP6 system and injected into Xenopus oocytes, the enzymatic activity was expressed in the oocytes. Second, the cloned cDNA was recombined to a vector under the control of the simian virus 40 early promoter and transfected to Chinese hamster ovary (CHO) cells. The enzymatic activity of the crude supernatant of transfected cells (CHO-HT2) reached to 400 pmol melatonin formed per min per mg of protein, which value was approximately 9% of that of bovine pineal supernatant. The amounts of enzyme protein estimated by immunoblotting were proportional to the enzymatic activity in both CHO and pineal gland. The content of HIOMT protein was 8- to 30-fold larger in pineal gland compared to CHO cells. On the other hand, the content of mRNA encoding the enzyme measured by dot hybridization with [32P]cDNA, was in the same range in both CHO cells and pineal glands. These data suggest that the 11-fold higher enzymatic activity in pineal gland is due to an accumulation of the enzyme protein, not to a high level of the mRNA and also indicate that the cloned cDNA can express an intact hydroxyindole O-methyltransferase enzyme in CHO cells.

Acetylserotonin O-Methyltransferase↗
PubMed1987

Cloning and sequence analysis of cDNAs for neurohypophysial hormones vasotocin and mesotocin for the hypothalamus of toad, Bufo japonicus.

The primary structures of the precursors of neurohypophysial hormones vasotocin (VT) and mesotocin (MT) in the hypothalamus of the toad Bufo japonicus were determined by analyzing the nucleotide sequences of the cloned cDNAs encoding them. The MT precursor consists of 125 amino acid residues containing a signal peptide followed directly by MT, which in turn is connected to the MT neurophysin by Gly-Lys-Arg, a processing and carboxyl-terminal amidation signal. In contrast, the VT precursor includes a glycoprotein of 36 amino acids following the VT neurophysin. Except for glycoprotein, the structures of MT and VT precursors are quite similar. RNA transfer blotting analysis showed that both MT and VT mRNAs are present in the brain but not in the liver, ovaries, and testes of the toad. The sequences and the structural organizations of the MT and VT precursors are highly homologous to those of their mammalian counterparts, oxytocin and arginine vasopressin precursors, respectively. This fact suggests that, in the evolutionary pathway of neurohypophysial hormones, VT is the ancestor molecule of vasopressin, while MT is that of oxytocin.

Amino Acid Sequence↗
PubMed1987

Monoclonal antibodies to and immunoaffinity purification of choline acetyltransferase from bovine brain.

Three hybridomas producing monoclonal antibodies to bovine brain choline acetyltransferase (ChAT) have been established by fusion of the spleen cells from a mouse immunized with purified enzyme with myeloma NS-1 cells. All three clones produced IgGl antibodies that reacted with enzyme protein denatured with sodium dodecyl sulfate. By using one of the monoclonal antibodies, a rapid and efficient immunoaffinity purification procedure of bovine ChAT has been established. Immunoblot analysis and immunoaffinity purification indicated that bovine ChAT is a single 68-kilodalton protein. The monoclonal antibodies will offer us a good tool to isolate the cDNA clones encoding ChAT.

Animals↗
PubMed1986

Cloning and sequence analysis of cDNA encoding urotensin I precursor.

The primary structure of the precursor of urotensin I, a neuropeptide hormone from the caudal neurosecretory system of the carp Cyprinus carpio, has been determined by analyzing the nucleotide sequence of cloned DNA complementary to the mRNA encoding it. The precursor consists of 145 amino acid residues and the carboxyl terminus represents the 41-amino acid sequence of urotensin I, preceded by Lys-Arg and followed by Gly-Lys. Sequence homology as well as similar organization of the precursors of urotensin I and mammalian corticotropin-releasing factors suggest that they are evolutionarily related. RNA transfer blot analysis indicates that mRNA encoding the precursor of urotensin I is present only in the spinal cord and not in the brain, intestine, liver, or kidney of the carp.

Amino Acid Sequence↗
PubMed1986

Cloning and sequence analysis of cDNAs encoding precursors of urotensin II-alpha and -gamma.

The primary structures of the precursors of urotensin II (UII)-alpha and -gamma, neuropeptide hormones of the caudal neurosecretory system of the carp, Cyprinus carpio, have been determined by analyzing the nucleotide sequences of cloned DNAs complementary to the mRNAs encoding them. A cDNA library, constructed with poly(A)+RNA in the preterminal spinal cord, was screened using 32P-labeled synthetic oligonucleotides representing all possible cDNA sequences corresponding to the pentapeptide common to all forms of carp UII (UII-alpha, -beta, and -gamma). Twenty out of 39 positive clones were analyzed with the restriction endonucleases, HindIII and PvuII, and classified into 4 groups. Nucleotide sequence analysis of 4 clones representing each group revealed that 2 clones encode the precursor of UII-alpha and the other 2 that of UII-gamma. Both precursors consist of 125 amino acid residues, and UII-alpha and -gamma exist at the carboxyl-termini preceded by Arg-Lys-Arg. The homology in both nucleotide and amino acid sequences between the precursors of UII-alpha and -gamma is more than 90%, suggesting that the genes were generated from a common ancestral gene by duplication. There is no sequence homology between the precursors of UII and urotensin I, another peptide hormone of the caudal neurosecretory system, nor between the precursors of UII and somatostatin-14. RNA transfer blot analysis indicated that mRNAs encoding the precursors of UII-alpha and -gamma are present in the spinal cord but not in the brain, intestine, liver, or kidney of the carp.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1985

[The clinicopathological effects of infusion of cis-platinum into the bronchial artery on lung cancer].

Ten patients with histologically and/or cytologically confirmed lung cancer were treated with cis-platinum (CDDP) infused into the bronchial artery (BAI). Radiological findings on chest X-rays taken two weeks after BAI were evaluated according to Takeuchi's criteria. The radiological results were judged to be quite effective in one case of squamous cell carcinoma, with more than 50% shrinkage of the tumor shadow, and to be effective in nine others, with more than 20% shrinkage. As for the histological effects, grade III in one case of squamous cell carcinoma and grade IIa in two cases of adenocarcinoma and one of large cell carcinoma were obtained by Shimosato's criteria. In conclusion, BAI with CDDP was sufficiently useful as a chemotherapeutic modality for lung cancer, especially squamous cell carcinoma, because higher local dosage of CDDP can be delivered without systemic toxicity.

Adenocarcinoma, Papillary↗
PubMed1985

[Results of surgical treatment for large cell carcinoma of the lung].

Twenty-six patients with histologically confirmed large cell carcinoma of the lung were evaluated. The overall five-year survival of these cases was 19.9%. Fifteen patients who had undergone surgical resection, excluding one operative death, had a five-year survival rate of 33.8%. None of the nonresected cases survived more than six months after the beginning of treatment. Among prognostic factors in surgical treatment of large cell carcinoma, survival correlated well with clinical stage, extent of lymph-node metastasis, curative resection and histological subtypes. As for histological subtypes, stratified subtype by division of nongiant cell carcinoma revealed poorer prognosis than that of nonstratified subtype and appeared to have some correlation with the frequency of vascular invasion by the tumor. From the clinical viewpoint, giant cell carcinoma should be regarded as a distinct clinical entity. Two cases, however, had benefit of long-term survival due to surgical modality. The clinicopathological analysis of those cases remains to be resolved in the future.

Adult↗
PubMed1984

Increase of choline acetyltransferase by colchicine in primary cell cultures of spinal cord.

Colchicine (5-10 microM) increased choline acetyltransferase (ChAT) activity 5-10-fold and suppressed acetylcholinesterase (AChE) and glutamate decarboxylase (GAD) activities to 30% and 50%, respectively, of the levels of control cells in mouse spinal cord cells cultured for several days. The synthesis of radiolabeled acetylcholine (ACh) from [14C]choline was also enhanced 4.6-fold, although the uptake of [14C]choline into cells was decreased to 80% of control level. Neither the incorporation of [3H]leucine into protein nor the total amount of protein was increased by colchicine. Vinblastine also increased ChAT activity while cytochalasin B was not effective. Immunochemical titration study revealed that the increase of ChAT activity by colchicine was due to the accumulation of ChAT molecules. Co-culture of spinal cord cells with skeletal muscle markedly stimulated ChAT activity, and the addition of colchicine to the cocultures showed greater than additive effect. These observations indicate that colchicine increases ChAT molecules in a specific manner, that the stimulatory effect of colchicine on ChAT activity is possibly mediated via the interaction with microtubules, and that the increase of ChAT activity is based on a mechanism different from that of co-cultures with skeletal muscle cells.

Acetylcholinesterase↗
PubMed1984

[Effects of infusion of cis-platinum into the bronchial artery in lung cancer].

5 patients with histologically and/or cytologically confirmed lung cancer were treated with Cis-platinum (CDDP) infused into the bronchial artery (BAI). 2 patients with squamous cell carcinoma disclosed hypervascular type and 3 patients with adenocarcinoma demonstrated avascular or hypovascular type on bronchial arterial angiography. All patients showed more than 20% shrinkage of the tumor shadow on chest X-ray taken 2 weeks after BAI and were evaluated to be radiologically effective by Takeuchi's criteria. One patient with squamous cell carcinoma who underwent lobectomy after BAI with 100 mg of CDDP showed grade III histological effects by Shimosato's criteria. The other patient had superior vena cava syndrome and showed regression of a tumor which occluded the orifice of the right upper and showed lobe after BAI with 100 mg of CDDP. By of contrast, only 1 out of the 3 patients with adenocarcinoma who under went resection showed a grade IIa histological effect after BAI with 50 mg of CDDP. Side effects included mild gastrointestinal toxicity in 1 out of the 5 patients. In conclusion, BAI with CDDP was shown to be sufficiently useful as a chemotherapeutic modality for lung cancer, especially for patients with squamous cell carcinoma, because higher local doses can be delivered without systemic toxicity.

Adenocarcinoma↗