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Biomedical subjects

H Kawauchi

Publications and source records attributed to H Kawauchi.

At least 55 records · Page 3Linked to original sources
PubMed1997

Sturgeon proopiomelanocortin has a remnant of gamma-melanotropin.

For the investigation of the evolution of the proopiomelanocortin (POMC) gene in early ray-finned fishes, nucleotide sequence of POMC cDNA from a chondrostean fish, the sturgeon has been determined. POMC cDNA was amplified by PCR from double-strand cDNA prepared from sturgeon pituitary and ligated with lambdaZAP II. The POMC cDNA consists of 1079 bp without a poly-A. An open reading frame of the POMC cDNA encodes 263 amino acid residues. Sturgeon POMC contains ACTH, alpha-melanotropin (MSH), beta-MSH and beta-END at positions (115-153), (115-127), (186-202) and (205-238), respectively. Location of POMC(51-72) is homologous to gamma-MSH, whereas the third residue of MSH-core sequence, His-Phe-Arg-Trp, is changed to His. Moreover, there are no basic amino acids to serve as a processing signal on the N-terminal side of POMC(51-72). These structural characteristics suggest that an ancestor of the ray-finned fishes had gamma-MSH, whereas significant mutations occurred during the evolution of chondrostean fish.

Adrenocorticotropic Hormone↗
PubMed1997

Isolation and characterization of two distinct gonadotropins from the pituitary gland of Mediterranean yellowtail, Seriola dumerilii (Risso, 1810).

Two gonadotropins, GTH I and GTH II, were isolated and chemically characterized from the pituitary of Mediterranean yellowtail. They were extracted with 35% ethanol-10% ammonium acetate, separated by ion-exchange chromatography on a DE-52 column, and purified by reversed-phase high-performance liquid chromatography on Asahipak C4P-50 and subsequently by gel filtration chromatography on Superdex 75. The molecular weights were estimated at 47 kDa for GTH I and 29 kDa for GTH II by SDS-PAGE and at 49 kDa for GTH I and 42 kDa for GTH II by gel filtration. GTH II was completely dissociated, while GTH I was partially dissociated into alpha- and beta-subunits by treatment with 0.1% trifluoroacetic acid. The complete amino acid sequences of GTH alpha-, GTH I beta-, and GTH II beta-subunits were determined. The GTH alpha-subunit consisted of 91 amino acid residues. The GTH I beta and GTH II beta consisted of 105 and 115 amino acid residues, respectively, and had a 28% sequence identity to each other. They had the highest sequence identity with the respective gonadotropin subunits of bonito, tuna, and striped bass: 81-83% for GTH alpha, 67-71% for GTH I beta, and 91-93% for GTH II beta. The sequence identity of the GTH alpha-subunit with those of other teleosts and human and bovine LH and FSH was 57-67%. The GTH I beta-subunit showed a low sequence identity with other known fish GTH I beta s (36-51%) and was more similar to human and bovine FSH beta s (34% identity) than to human and bovine LH beta s (29% identity). The sequence identity of the GTH II beta-subunit with those of other teleosts was higher (60-73%), being more similar to LH beta s (43% identity) than FSH beta s (38% identity). Thus, two distinct gonadotropins, GTH I and GTH II, homologous to mammalian FSH and LH, respectively, are synthetized by M. yellowtail pituitary glands.

Amino Acid Sequence↗
PubMed1997

Salmon thyroid-stimulating hormone: isolation, characterization, and development of a radioimmunoassay.

Coho salmon (Oncorhynchus kisutch) thyroid-stimulating hormone (TSH) was isolated by ethanol extraction of pituitary glands from mature coho salmon. Extraction was followed by gel-filtration chromatography on Sephadex G-100 superfine, anion-exchange chromatography on a Whatman DE-52 column, and finally by reverse-phase high-performance liquid chromatography. Fractions were monitored for TSH content by a homologous in vivo bioassay and by immunoblots using anti-human TSH beta-subunit antisera. In vivo treatment of coho salmon parr with coho salmon TSH caused a dose-dependent increase in plasma thyroxine level similar to that induced by bovine TSH. The N-terminal sequence (25 residues) of the salmon TSH beta subunit has 56% sequence identity to that of human TSH beta subunit and is identical to the deduced amino acid sequence of trout TSH beta subunit. The N-terminal sequence (25 residues) of the salmon TSH alpha subunit is identical to gonadotropin alpha-II subunit. Molecular sizes of the alpha and beta subunits are 18,000 and 24,000 daltons, respectively, as estimated by SDS-PAGE. Antiserum generated against salmon TSH, which was preadsorbed with alpha subunit using an alpha-subunit affinity column, detected only salmon TSH beta subunit by immunoblot and specifically stained thyrotropin-producing cells of the pituitary gland. A homologous radioimmunoassay (RIA) was developed using purified salmon TSH standard, iodinated TSH beta subunit, and antiserum generated against salmon TSH. Cross-reactivities of GTH I, GTH II, GTH I beta and GTH II beta subunits, alpha subunit, growth hormone, prolactin, and somatolactin were less than 1%. Displacement curves for serial dilutions of plasma and pituitary extracts of various salmonid species, as well as coho salmon pituitary cell culture medium, were parallel to the coho salmon TSH standards. In contrast, plasma of hypophysectomized juvenile coho salmon and pituitary extracts of Pacific tomcod (Microgadus proximus) did not displace bound radiolabeled salmon TSH. Finally, in vivo injection of juvenile coho salmon with triiodothyronine decreased plasma TSH levels, whereas the goitrogen, methimazole, increased plasma TSH levels. Injection of gonadotropin-releasing hormone agonist did not alter plasma TSH. These data suggest that the RIA is specific for TSH and confirm a negative-feedback relationship between the thyroid hormones and TSH.

Amino Acid Sequence↗
PubMed1997

Immunostimulating effect of growth hormone: in-vivo administration of growth hormone in rainbow trout enhances resistance to Vibrio anguillarum infection.

In-vivo administration of purified (nGH) and recombinant (rGH) chum salmon, Oncorhynchus keta, growth hormone enhanced the survival of rainbow trout, O. mykiss, against virulent Vibrio anguillarum. Macrophages obtained from rainbow trout that were treated in vivo with either GH preparation showed enhanced stimulation of the chemiluminescent responses induced by opsonized V. anguillarum cells and increased phagocytic activities. However, the serum bactericidal activity against V. anguillarum was not enhanced in fish injected with nGH or rGH. These findings show that in-vivo administration of either GH preparation can effectively prime macrophages and increase the resistance to V. anguillarum in rainbow trout.

Adjuvants, Immunologic↗
PubMed1997

[Investigation of the usefulness of CYFRA 21-1 as a tumor marker in squamous cell carcinomas of the head and neck].

It has been demonstrated that CYFRA 21-1 (ELISA kit), which recognizes the soluble cytokeratin 19 fragment, is useful for assessing circulating tumor antigens in sera of patients with lung cancer. In this study, we compared the clinical significance of this new marker with the established squamous cell carcinoma antigen (SCC Ag), using sera from patients with head and neck malignant disease, healthy controls, and supernatants of established cell lines derived from squamous cell carcinomas and adenocarcinomas. The subjects were: Group A, 39 patients with malignant disease of the head and neck. Group B, 11 patients considered to be tumor-free after treatment. Group C, 67 patients with benign disease or healthy volunteers. Culture supernatants: 11 cell lines established from squamous cell carcinomas and adenocarcinomas. Serum levels of CYFRA 21-1 and SCC Ag of group A were significantly higher than those of group C. This finding suggests that CYFRA 21-1 is useful as a tumor marker as well as SCC Ag. CYFRA 21-1 and SCC Ag levels of patients in group A at the early and progressive stages of disease were comparable to the levels in group C. Both tumor markers are therefore useful for diagnosis of in the early stage of cancer. We attempted to set a cut-off level of CYFRA 21-1. The sensitivity of CYFRA 21-1 is higher than that of SCC Ag, especially in patients in the early stage of the disease. This finding indicates that the CYFRA 21-1 is preferable to SCC Ag as a tumor marker for the diagnosis of patients even in the early stages of malignant disease. The levels of CYFRA 21-1 in culture supernatants derived from tumor cell lines are higher than those of SCC Ag in all cell lines. The levels of CYFRA 21-1 are measurable, with levels varying with the cell line. There appears to be no correlation between the level of CYFRA 21-1 and the character of the cell lines, but this issue remains to be further investigated.

Adult↗
PubMed1996

Complete amino acid sequences of follitropin and lutropin in the ostrich, Struthio camelus.

We determined the complete amino acid sequences of two pituitary gonadotropins, follitropin and lutropin in the ostrich, thereby providing the first information on the structure of avian follitropin. Ostrich follitropin and lutropin both consist of two subunits: a common alpha-subunit and a hormone-specific beta-subunit. The alpha-subunit is composed of 96 amino acid residues and has 70-80% sequence identity with the alpha-subunits of most vertebrates. The ostrich follitropin beta-subunit consists of 106 amino-acid residues, and shows 70-74% sequence identity with mammalian follitropins beta, 61% with amphibian follitropin beta, 39-46% with teleost gonadotropins II beta and 32-44% with teleost gonadotropins I beta. The ostrich lutropin beta-subunit consists of 128 amino-acid residues, and exhibits 76-78% sequence identity with other avian lutropins beta, 44-50% with teleost gonadotropins II beta, 45% with amphibian lutropin beta, 41-44% with mammalian lutropins beta, and 25-36% with teleost gonadotropins I beta. Sequence comparison revealed that lutropin beta-subunits are more class-specific and have diversified approximately twice as fast follitropin beta-subunits, although segments essential for maintaining higher-order structures have been conserved.

Amino Acid Sequence↗
PubMed1996

In vitro activation of fish phagocytic cells by GH, prolactin and somatolactin.

The activation of rainbow trout (Oncorhyndus mykiss) phagocytic cells by chum salmon GH, prolactin (PRL) and somatolactin (SL) was investigated in vitro. Rainbow trout kidney leucocytes were cultured in RPMI 1640 medium containing 1, 10, 50 or 100 ng of each hormone/ml and the production of superoxide anion was measured using reduction of nitroblue tetrazolium (NBT) and ferricytochrome C. Macrophages incubated with 10-100 ng GH or PRL/ml showed significantly enhanced production of superoxide anion in both the NBT and ferricytochrome C tests compared with control macrophages (without hormone). SL did not induce enhanced production of superoxide anion in macrophages. The phagocytic cells treated with GH or PRL also showed increased phagocytic activity and phagocytic index. However, the cells treated with SL showed no enhancement of phagocytic activity or index. These results indicate that GH and PRL stimulate macrophages in vitro.

Animals↗
PubMed1995

Melanotropin and corticotropin are encoded on two distinct genes in the lamprey, the earliest evolved extant vertebrate.

In the lamprey, which is a member of the oldest extant class of vertebrates, the agnathans, melanotropins (MSH) and corticotropin (ACTH) were found to be encoded on two distinct genes. In all other vertebrates, a single precursor gene, proopiomelanocortin (POMC), encodes MSH and ACTH, as well as beta-endorphin (END). Two different cDNAs were cloned from a lamprey pituitary lambda gt11 cDNA library using antisera against lamprey MSH-B and ACTH(1-16). One cDNA encoded MSH-B, MSH-A and beta-END, while the other cDNA encoded nasohypophysial factor (NHF), ACTH and a different beta-END, but not MSH-A and MSH-B. Northern blot analysis demonstrated that in the adult lamprey pituitary, genes for MSH and ACTH are expressed in the pars intermedia (PI) and pars distalis (PD), respectively.

Adrenocorticotropic Hormone↗
PubMed1995

The complete amino acid sequence of chum salmon stanniocalcin, a calcium-regulating hormone in teleosts.

Stanniocalcin (STC) is a hypocalcemic hormone secreted from the corpuscles of Stannius of bony fish. Chum salmon STC was isolated in pure form by ion-exchange chromatography and reversed phase high performance liquid chromatography following ethanol-ammonium extraction of the tissues. A cDNA was cloned from cDNAs of the tissue by the PCR method using two primers corresponding to the N- and C-terminal amino acid sequence of the hormone. Sequence analysis of the protein and the cDNA revealed that chum salmon STC is a homodimer, and that the monomer consists of 179 amino acids including 11 half-Cys residues and one N-linked glycosylation site, which is 44 residues smaller at the C-terminal region than the sequence deduced from coho salmon STC cDNA.

Amino Acid Sequence↗
PubMed1995

Distribution of lamprey adrenocorticotropin and melanotropins in the pituitary of the adult sea lamprey, Petromyzon marinus.

Three peptides containing the melanotropin (MSH)-core sequence, YXMXHFRWG, have been isolated recently from the pituitary glands of adult sea lampreys (Petromyzon marinus) and were tentatively assigned as lamprey adrenocorticotropin (ACTH), MSH-A, and MSH-B, respectively. Both MSHs differed significantly from gnathostome MSHs and cannot be assigned as alpha-MSH, beta-MSH, or gamma-MSH. The aim of the present study was to localize these peptides in the lamprey pituitary using antisera generated against synthetic lamprey ACTH1-16, MSH-A, and MSH-B. ACTH-like immunoreactivity was found in most cells of the rostal pars distalis (RPD) and in a few scattered cells of the proximal pars distalis (PPD). MSH-A-like immunoreactivity was found in most cells of the RPD, a few scattered cells of the PPD, and almost all cells of the pars intermedia (PI). MSH-B-like immunoreactivity was found only in the PI, where almost all cells were stained. Thus, the topographic distributions of ACTH and MSHs in the lamprey pituitary were similar to those in gnathostome vertebrates.

Adrenocorticotropic Hormone↗
PubMed1995

Complete amino acid sequence of crocodile growth hormone.

This paper describes the isolation and complete amino acid sequence of growth hormone (GH) from the pituitary gland of the crocodile, Crocodylus novaeguineae. GH was detected in an alkaline extract of the pituitary by specific immunoblot reactivity with rabbit antisera against cattle and salmon GH and isolated in pure form by DEAE-cellulose chromatography and by reversed-phase high-performance liquid chromatography. The yield of the GH was 2.2 mg from 3.5 g of wet tissue. Crocodile GH consists of 190 residues and is highly similar to chicken, duck, and sea turtle GHs with 90-92% structural similarity, although it shows only 27% identity with crocodile prolactin. A molecular phylogenetic tree of GH from 39 vertebrate species is proposed on the basis of sequence comparison.

Alligators and Crocodiles↗
PubMed1995

Isolation and characterization of [Pro2]somatostatin-14 and melanotropins from Russian sturgeon, Acipenser gueldenstaedti Brandt.

A new form of somatostatin (SRIH), along with melanotropins (MSHs), was isolated from pituitaries of the Russian sturgeon Acipenser gueldenstaedti Brandt by gel filtration, ion exchange, and reversed-phase HPLC following acid-acetone extraction. The sturgeon SRIH consists of 14 amino acid residues and differs from mammalian SRIH-14 by the substitution Pro for Gly at position 2. Synthetic [Pro2]SRIH-14 was as potent as mammalian SRIH-14 in inhibiting release of growth hormone into medium from the organ-cultured pituitary of rainbow trout. Sturgeon alpha-MSH has the same amino acid sequence as those found in mammals. Sturgeon beta-MSH is composed of 17 amino acid residues, and its amino acid sequence is identical to the N-terminal 15 residues of salmon beta-MSH I and to the C-terminal 2 residues of mammalian beta-MSH.

Adrenocorticotropic Hormone↗
PubMed1995

Immunocytochemical localization of IGF-I in the ovary of the red seabream, Pagrus major.

There are many biochemical and molecular biological data indicating that IGF-I is present in teleost fish. The present study examines red seabream for the presence and cellular distribution of IGF-I in the ovarian follicles during oocyte growth and final maturation of oocytes by immunocytochemistry. Immunoreactivity appeared in the granulosa cells at the lipid stage. Immunoreactivity became strong at the primary yolk globule stage and diminished in intensity during development of maturational competence (responsiveness to maturation-inducing steroid) and final maturation of oocytes. Intense immunoreactivity was also localized in the outer layer of the zona radiata and the peripheral region of the ooplasm at the primary yolk globule stage. Postovulatory follicles had low immunoreactivity in the early phase of degeneration. These immunocytochemical observations show that the granulosa cell layer is the main site of IGF-I production. IGF-I may be involved in granulosa cell proliferation and differentiation.

Animals↗
PubMed1995

A novel glycoprotein in the olfactory and pituitary systems of larval and adult lampreys.

A novel homodimeric glycoprotein was isolated and characterized from the pituitaries of adult sea lampreys, Petromyzon marinus, modern representatives of the earliest vertebrates. The monomer consists of 121 amino-acid residues in a sequence that has no resemblance to any known pituitary hormone. Whereas this protein is localized in most cells of the rostral pars distalis of adult lampreys, we have chosen to name it, nasohypophysial factor (NHF), because it first appears in the olfactory system of developing larval lampreys. Not only may NHF be a new pituitary hormone but a useful probe for examining the ontogenetic and phylogenetic relationships of the pituitary and olfactory systems in vertebrates.

Amino Acid Sequence↗
PubMed1995

Effect of oxatomide on otitis media with effusion--an experimental study.

The therapeutic effect of oxatomide on experimental otitis media with effusion (OME) induced by secondary immune response was investigated in chinchillas. Oxatomide was administered daily in three different doses (5, 10, 30 mg/kg b.w.t.) for a period of 2 weeks, starting 2 days after the instillation of keyhole limpet hemocyanin (KLH) into the tympanic cavity. The severity of OME was assessed by observing the eardrum under a surgical microscope, by tympanography, and by histological findings. The concentrations of leukotriene C4 and LTD4 in middle ear effusion (MEE) were also measured, 7 days after the KLH instillation. It was found that oral administration of oxatomide in dosages of 10 and 30 mg/kg alleviated the experimental OME. The mean LTD4 concentration measured in MEE from chinchillas given 30 mg oxatomide was significantly lower than that measured in the control group. The radioactivity of 14C-labelled oxatomide in MEE and the autoradiographic findings revealed that orally administered oxatomide was rapidly absorbed from the digestive tract and transported to the middle ear via blood circulation. The findings suggest that the effect of oxatomide can be anticipated in patients with OME.

Acoustic Impedance Tests↗
PubMed1995

Somatolactin, a novel pituitary protein: isolation and characterization from Sparus aurata.

The complete amino acid sequence of somatolactin, a new pituitary protein belonging to the growth hormone/prolactin family, from the gilthead sea bream Sparus aurata has been determined. Somatolactin was isolated from the pituitary by alkaline extraction, gel filtration on a Sephadex G-100 column, and reversed-phase high-performance liquid chromatography (rpHPLC) on a TSK gel ODS-120T column. The purified protein was confirmed to be somatolactin by immunoblotting using chum salmon somatolactin antisera. It was found that Sparus aurata somatolactin consists of two forms; one form (28 kD) is probably a glycosylated form, while the other (25 kD) is a simple protein form, as was found also in Atlantic cod. The somatolactin consists of 207 amino acids that show remarkable conservation among fish somatolactins.

Amino Acid Sequence↗
PubMed1994

Cloning and nucleotide sequence of a bacterial cytochrome P-450VD25 gene encoding vitamin D-3 25-hydroxylase.

The gene encoding an enzyme that catalyzes the hydroxylation at position 25 of vitamin D-3 was cloned from an actinomycete strain, Amycolata autotrophica, by use of a host-vector system of Streptomyces lividans. The amino acid sequence deduced from the nucleotide sequence revealed that this enzyme, tentatively named P-450VD25, contains several regions of strong similarity with amino acid sequences of cytochromes P-450 from a variety of organisms, primarily in the regions of an oxygen-binding site and a heme ligand pocket. Especially, P-450VD25 shows end-to-end similarity in amino acid sequence to P-450dNIR of Fusarium oxysporum and P-450SU2 of Streptomyces griseolus. The recombinant S. lividans strain containing the P-450VD25 gene on a multicopy plasmid converted vitamin D-3 in the medium into 25-hydroxyvitamin D-3 at a maximum yield of 10%.

Actinomycetales↗