Search PubMed⌕ Search

Biomedical subjects

N Ueno

Publications and source records attributed to N Ueno.

At least 289 records · Page 16Linked to original sources
PubMed1986

Isolation of an amino terminal extended form of basic fibroblast growth factor.

Extraction of bovine pituitaries in the presence of enzyme inhibitors (2 mM PMSF, 2 mM sodium tetrathionate, 15 microM pepstatin A, and 1 mM EDTA) resulted in the isolation of two distinct forms of basic fibroblast growth factor. Partial characterization of both molecules showed one form to be identical to basic FGF(1-146) which has already been reported by our laboratory. The second form was estimated by SDS-PAGE to have a molecular weight of 17,000 Daltons which is slightly larger than that of basic FGF(1-146). Amino acid analysis shows the presence of 8 new residues more than basic FGF(1-146) which accounts for the difference in molecular weight. Gas-phase sequencing of this molecule indicated that it bears a blocked amino terminus. Furthermore, this higher molecular weight form of basic FGF did not show immunoreactivity with antibodies specific for the amino terminus of basic FGF(1-146) but cross reacted with antibodies generated against midportion fragments of basic FGF(1-146), indicating that the molecule is amino terminally extended. Like basic FGF(1-146), the molecule is a potent mitogenic factor for vascular endothelial cells. Taken together these results demonstrate the existence of a precursor form of basic FGF which is extended by 8 residues at the amino terminus with the first residue being blocked.

Amino Acids↗
PubMed1986

Inhibin and beta type transforming growth factor (TGF beta) have opposite modulating effects on the follicle stimulating hormone (FSH)-induced aromatase activity of cultured rat granulosa cells.

We have recently observed that attomolar concentration of exogenously added TGF beta, a molecule structurally related to inhibin, can stimulate the basal secretion of FSH in a pituitary cell culture. Inhibin purified from porcine follicular fluid antagonizes this activity of TGF beta. To understand further the homeostatic regulatory properties of inhibin and TGF beta we have investigated whether the aromatase activity of ovarian granulosa cells is also subject to intra-ovarian modulation by these peptides. Granulosa cells from immature hypophysectomized diethylstilbestrol-treated rats were cultured for 2 days with androstenedione (10(-7) M) as a substrate, oFSH (2 ng), and different amounts of TGF beta or inhibin. Basal estrogen secretion was negligible and remained unaffected by treatment with purified TGF beta or inhibin (10 ng/ml), whereas treatment with oFSH (2 ng/ml) produced a 100-fold increase in estrogen accumulation. The concurrent application of increasing concentrations (10 pg-10 ng/ml) of TGF beta produced dose-dependent increments in the FSH-stimulated accumulation of estrogen with a ED50 of 0.3 +/- 0.02 ng/ml. On the other hand, concurrent incubation of FSH with inhibin ranging from 10 pg to 10 ng/ml decreases the FSH-mediated estrogen secretion. TGF beta antagonizes the inhibition of inhibin on aromatase activity. These findings suggest that inhibin and TGF beta, two closely related molecules, play novel and opposite roles in modulating the follicular functions.

Androstenedione↗
PubMed1986

Type beta transforming growth factor (TGF-beta) is a potent stimulator of the basal secretion of follicle stimulating hormone (FSH) in a pituitary monolayer system.

In view of striking similarities between TGF-beta and inhibin, we investigated the possibility that TGF-beta might modulate pituitary hormone release in vitro. Long term incubations of beta transforming growth factor (TGF-beta) with rat anterior pituitary cells for 48 hr stimulates the basal secretion of FSH in a dose-dependent manner. The secretion of LH, TSH, GH, ACTH and PRL is not modified by TGF-beta. The minimal effective concentration of TGF-beta is 10 pg/ml (less than 500 attomolar) and is dose dependent over a range from 1 pg to 10 ng/ml. Treatment of cells with TGF-beta for short incubation times (4 hr) in assays similar to that used for hypophysial releasing factors is not effective, indicating that TGF-beta acts through a cellular mechanism distinct from that of LRF. Inhibin-A, recently characterized on the basis of its capacity to specifically inhibit the secretion of FSH in the 48 hr bioassay system inhibits the stimulatory effect of TGF-beta on FSH-release. Analyses of the dose response curves indicate that the interaction occurs in a typical non-competitive manner. The results suggest that a TGF-beta-like molecule, present in follicular fluid, may be responsible for the FSH-releasing activity ("anti-inhibin" activity) observed by us and others during the process of isolating inhibin from follicular fluids. They also suggest an important role for inhibin and the TGF-beta related molecules in modulating pituitary gonadotropin release.

Animals↗
PubMed1986

Impaired monocyte function in glycogen storage disease type Ib.

Phagocyte (neutrophil and monocyte) function was evaluated in a boy with glycogen storage disease type Ib. Neutrophils were found to be defective in motility and respiratory burst and monocytes showed a defect in respiratory burst but not in motility. These results suggested that the glucose-6-phosphate transport system plays a role in the function of neutrophils and monocytes and that these two phagocytes are different from each other in their energy metabolism for motility.

Adolescent↗
PubMed1986

Fibroblast growth factor promotes survival of dissociated hippocampal neurons and enhances neurite extension.

Basic fibroblast growth factor (FGF) has been found to increase neuronal survival and neurite extension in a highly purified population of fetal rat hippocampal neurons under well-defined serum-free cell culture conditions. In the presence of FGF, neuronal survival after 7 days in culture on a simple plastic substrate is increased 4-fold, to 54% of the initial population. Survival is increased 2-fold to 40% on polyornithine-laminin. When FGF was bound to plastic or heparin substrates, neurite outgrowth was significantly increased to lengths comparable to those seen with laminin; however, FGF produced no further increase in neurite outgrowth on laminin. Half-maximal survival was observed at FGF concentrations of about 15 pg/ml (1 pM); half-maximal process outgrowth occurred at about 375 pg/ml (20 pM). The responsive cells were identified as neurons by their labeling with tetanus toxin and by antibodies to neurofilaments and to the neuron-specific enolase. Astrocytes, identified by the presence of glial fibrillary acidic protein, constituted about 10% of cells present at 1 week both in the presence and in the absence of FGF. These results strongly suggest that, in addition to its known mitogenic effects on nonneuronal cells, FGF possesses neurotrophic activity for hippocampal neurons.

Animals↗
PubMed1986

[Clinical evaluation of kallikrein in male infertility].

Kallikrein was orally administered to 44 patients with male infertility at a dose of 90 or 120 KU/day for three months. The sperm density, sperm motility and fertility index (F.I.) were improved in 51.1%, 37.2% and 38.9% of the treated patients, respectively. Statistical analysis showed that significant improvement was noted in sperm density and F.I. between before and after the treatment, but not in sperm motility, sperm volume nor sperm morphology. During the period of treatment, in these treated 44 patients, 4 patients (9.1%) succeeded in pregnancy.

Administration, Oral↗
PubMed1986

[Chemiluminescence of whole blood--analysis of the kinetics and application of clinical examination of phagocytic functions of whole blood from various type of diseases].

Luminol-dependent chemiluminescence of whole blood (whole blood CL) was developed to estimate the phagocytic function of granulocytes and the serum opsonin activity simultaneously. Whole blood (0.1 ml) was examined directly and results were obtained within 20 minutes. Phagocytic function of granulocytes can be estimated from the peak CL of whole blood and the number of granulocytes in a specimen, and the opsonin activity from the amount of time the peak CL is shown after the addition of nonopsonized CL inducer (nonopsonized zymosan). Subsequently, whole blood CL was measured to evaluate phagocytic functions in children with disease. Patients with chronic granulomatous disease showed no CL, and one of their mothers (1/3) showed low CL, suggesting she is a carrier. Two patients with hypocomplementemia (SLE and chronic nephritis) showed low serum opsonin activity, and phagocytic function of their granulocytes was enhanced. In cord blood and newborn infants serum opsonin activity was low, and phagocytic function of granulocytes was slightly decreased in cord blood but not in newborn infants. Patients with systemic bacterial infections showed an increased phagocytic function of granulocytes. Anti-cancer drugs decreased serum opsonin activity in children with leukemia or lymphoma. The children treated with L-asparaginase had very low opsonin activity, suggesting the drug inhibits complement synthesis. The measurement of whole blood CL was useful for monitering the phagocytic functions of blood after granulocyte transfusion.

Bacterial Infections↗
PubMed1986

Effect of adriamycin entrapped by sulfatide-containing liposomes on ovarian tumor-bearing nude mice.

Sulfatide-containing liposomes showed the highest degree of adriamycin entrapment of all the liposomes tested. Adriamycin was bound to the sulfatide anions on the liposomal membrane, inserted into the membrane, and incorporated into the aqueous compartment of the vesicle. Liposome-entrapped adriamycin was maintained at a much higher blood level than free adriamycin, and reached a lower concentration in the heart than did the free drug, which might lead to lower cardiotoxicity of the drug. Incorporation of adriamycin into ovarian tumor transplanted into nude mice was increased when entrapped by the sulfatide-containing liposomes. Liposome-entrapped adriamycin did not induce the drastic loss of body weight which occurred with the free drug. The growth of ovarian tumor was inhibited by liposome-entrapped adriamycin to the same degree as free adriamycin. Having these advantages, sulfatide-containing liposomes could be useful carriers of adriamycin for cancer chemotherapy.

Animals↗
PubMed1985

Primary structure of bovine brain acidic fibroblast growth factor (FGF).

The major anionic mitogenic polypeptide for endothelial cells, acidic fibroblast growth factor (FGF), has been purified to homogeneity from bovine brain and its complete primary structure established by gas-phase sequence analysis. The 140 amino acid (Mr 16,000) protein has been previously shown to be a potent growth factor for many diverse cell types of mesodermal origin, in vitro, and an angiogenic agent, in vivo. The amino acid sequence of bovine brain acidic FGF has a 53% absolute homology with that of bovine pituitary basic FGF suggesting that these endothelial cell mitogens are derived from a single ancestral gene.

Amino Acid Sequence↗
PubMed1985

Identification of plasma inactive renin as prorenin with a site-directed antibody.

A sequence-specific antibody that recognizes a portion of the prosegment of human renin precursor was raised and used to provide direct evidence that plasma inactive renin contains the prosequence of renal renin and is therefore probably prorenin rather than an inactivated form of previously active renin. The information may help not only to resolve a major controversy concerning the nature of inactive renin in human plasma but also to elucidate its exact physiological role.

Antibody Formation↗
PubMed1985

A nonmitogenic pituitary function of fibroblast growth factor: regulation of thyrotropin and prolactin secretion.

The addition of fibroblast growth factor (FGF) to primary cultures of rat anterior pituitary cells modifies their response to thyrotropin-releasing factor in a dose-dependent manner. While the pituitary response to the other releasing factors (corticotropin-releasing factor, growth hormone-releasing factor, and gonadotropin-releasing factor) is not altered, FGF increases both the sensitivity of the cells to thyrotropin-releasing factor and the amounts of prolactin and thyrotropin released. A minimum of 24 hr of preincubation with FGF is required to modify the pituitary response, and maximal effects were observed with 48 and 72 hr of preincubation. The effective doses of FGF are similar to those described for its mitogenic activity (i.e., 1-100 pM), but inhibition of cell growth with 5-fluorodeoxyuridine does not modify the effect of FGF on thyrotropin and prolactin release. These results suggest a novel paracrine, if not autocrine, role of pituitary FGF in the homeostatic mechanisms that regulate the secretion of prolactin and thyrotropin. They also suggest that the biological significance of the presence of FGF in various tissues may not be directly related to its in vitro mitogenic activity.

Animals↗
PubMed1985

Primary structure of bovine pituitary basic fibroblast growth factor (FGF) and comparison with the amino-terminal sequence of bovine brain acidic FGF.

The two major mitogenic polypeptides for endothelial cells have been purified to homogeneity. The complete primary structure of bovine pituitary basic fibroblast growth factor (FGF) and the amino-terminal amino acid sequence of bovine brain acidic FGF have been established by gas-phase sequence analyses. Homogeneous preparations of these polypeptides are potent mitogens (basic FGF, ED50 approximately equal to 60 pg/ml; acidic FGF ED50 approximately equal to 6000 pg/ml) for many diverse cell types including capillary endothelial cells, vascular smooth muscle cells, and adrenocortical and granulosa cells; in vivo, basic FGF is a powerful angiogenic agent in the chick chorioallantoic membrane assay. The available protein sequence data demonstrate the existence of significant structural homology between the two polypeptides.

Amino Acid Sequence↗
PubMed1985

Isolation and partial characterization of a Mr 32,000 protein with inhibin activity from porcine follicular fluid.

A Mr 32,000 protein with inhibin activity was isolated from porcine follicular fluid by heparin-Sepharose affinity chromatography, gel filtration on Sephacryl S-200, and four reversed-phase HPLC steps. The isolated molecule is composed of two chains having molecular weights of 18,000 and 14,000, respectively, and bound together by disulfide bonds. Amino acid sequence analysis revealed the 10 NH2-terminal residues of the Mr 18,000 chain to be Ser-Thr-Ala-Pro-Leu-Pro-Trp-Pro-Trp-Ser- and those of the Mr 14,000 chain to be Gly-Leu-Glu-Xaa-Asp-Gly-Arg-Thr-Asn-Leu-. This Mr 32,000 protein specifically inhibits the basal secretion of FSH, but not that of LH, in the rat anterior pituitary monolayer culture system, with a half-maximal effective dose of 450 pg/ml.

Amino Acid Sequence↗