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Biomedical subjects

H Nagase

Publications and source records attributed to H Nagase.

At least 271 records · Page 15Linked to original sources
PubMed1995

Antagonistic effect of buprenorphine on the antitussive effect of morphine is mediated via the activation of mu 1-opioid receptors.

The effect of buprenorphine on the antitussive effect of morphine was examined in mice. Buprenorphine at doses of 0.1, 0.3 and 1 mg/kg given i.p. alone have no effects on the % inhibition in the number of capsaicin-induced coughs. However, pretreatment with the same doses of buprenorphine for 2 hr significantly attenuated the antitussive effect of morphine (3 mg/kg, i.p.). Naloxonazine, a selective mu 1-opioid receptor antagonist, had no effect of buprenorphine on antitussive effect of morphine. These results suggest that buprenorphine antagonizes the antitussive effect of morphine via the activation of mu 1-opioid receptors.

Animals↗
PubMed1995

Effect of naltrindole on the development of physical dependence on morphine in mice: a behavioral and biochemical study.

The effect of pretreatment with a delta opioid receptor antagonist, naltrindole (NTI), on the development of physical dependence on morphine was investigated in mice. Several withdrawal signs, an increase in cortical noradrenaline (NA) turnover and a decrease in dopamine (DA) turnover in the limbic forebrain were observed following naloxone challenge in morphine-dependent mice. Pretreatment with NTI (0.3-5 mg/kg, s.c.) during chronic morphine treatment dose-dependently suppressed the behavioral and biochemical changes after withdrawal. The blocking effects of NTI suggest that delta opioid receptors may play a significant role in modulating the development of physical dependence on morphine.

Animals↗
PubMed1995

Matrix metalloproteinases as insulin-like growth factor binding protein-degrading proteinases.

Insulin-like growth factor (IGF) bioavailability is modulated by specific IGFBPs, six of which are known (IGFBPs 1-6). Since IGFBPs have equal or higher affinity for IGFs than do IGF receptors, it is believed that degradation of IGFBPs by IGFBP-degrading proteinases is an important step in regulating IGF bioactivity. Recent studies from our laboratory have demonstrated that at least two IGFBPs, i.e. IGFBP-3 and -5, are degraded under physiologic conditions by matrix metalloproteinases (MMPs). In vitro, we have demonstrated that IGFBP-3 is degraded in human dermal fibroblast cultures by MMPs using a variety of techniques, including proteinase inhibition with a specific inhibitor of MMPs, i.e. tissue inhibitor of metalloproteinases (TIMP-1), immunoabsorption with specific antisera to human MMPs and a unique method developed in our laboratory, IGFBP-3 substrate zymography. Using similar methods, we have also demonstrated that MMPs, along with an unidentified 97-kDa proteinase, degrade IGFBP-5 in murine osteoblast cultures. In rat pregnancy serum, we have shown that degradation of IGFBP-3 is associated with MMP activity present in the serum, which likely arises from the placental compartment. Analysis of the cleavage products of IGFBP-3 produced by MMPs 1, 2 and 3 reveals that MMPs cleave exclusively within the non-homologous, mid-region of the molecule. Together, these studies suggest that MMPs, beyond their previously described roles as extracellular matrix degrading enzymes, may also exert effects on cellular growth and differentiation via degradation of IGFBPs.

Animals↗
PubMed1995

Co-culture of human breast adenocarcinoma MCF-7 cells and human dermal fibroblasts enhances the production of matrix metalloproteinases 1, 2 and 3 in fibroblasts.

No measurable amounts of matrix metalloproteinases (MMPs) were produced by human breast adenocarcinoma cell lines MCF-7 and BT-20 in culture. When MCF-7 cells were co-cultured with human dermal fibroblasts enhanced production of precursors of MMP-1 (interstitial collagenase), MMP-2 (gelatinase A), MMP-3 (stromelysin 1) and tissue inhibitor of metalloproteinase type 1 (TIMP-1) was observed. Immunohistochemical studies indicated that these pro-MMPs originated primarily from the fibroblasts, suggesting that MCF-7 cells have a stimulatory effect on stromal cells to produce at least three pro-MMPs and TIMP-1. BT-20 cells also enhanced the production of pro-MMP-2 and TIMP-1 in the dermal fibroblasts, but not of pro-MMP-1 and pro-MMP-3. Normal mammary epithelial cells promoted only TIMP-1 production. To investigate further the stimulatory factors from MCF-7 cells, the conditioned medium and the cell membrane were prepared and examined. The cell membrane fraction enhanced the production of pro-MMP-1 and -3 and TIMP-1, but not of pro-MMP-2. The conditioned medium, on the other hand, augmented the production of all four proteins in the fibroblasts. These observations suggest that breast adenocarcinoma MCF-7 cells in culture produce both soluble and membrane-bound factor(s) which stimulate the production of pro-MMPs and TIMP-1 in neighbouring stromal cells, but the factor(s) released into the medium and that associated with cell membranes are probably different. Such communication between the normal and malignant cell types may, in part, assist the cancer cells to invade and metastasise.

Adenocarcinoma↗
PubMed1995

Distinct genetic loci control development of benign and malignant skin tumours in mice.

Genetic susceptibility to chemically induced skin cancer in mice is controlled by multiple unlinked genetic loci. Mus spretus mice have dominant resistance genes which confer resistance to interspecific F1 hybrids with susceptible Mus musculus strains. We have mapped three major resistance loci using a combination of Mapmaker/QTL analysis and multiple regression analysis to mouse chromosomes 5 and 7. At least two independent loci on chromosome 7 exert their effects primarily during benign tumour development and have very little influence on tumour progression. On the other hand, probably a single locus on chromosome 5 affects both early and late stages of malignancy. The results indicate that benign and malignant tumours are largely under independent genetic control.

Animals↗
PubMed1995

X-ray microprobe analysis of corpora amylacea.

Since corpora amylacea is concentrated in the high density fraction in the subcellular fractionation of autopsy brain. It is suspected that inorganic materials accumulate in corpora amylacea. Therefore, elemental analyses of partially purified corpora amylacea from autopsy brain from a patient with Alzheimer's disease and those from brain of a non-demented patient were performed by the X-ray microprobe method. Prominent peaks of sodium, phosphorus, sulphur and chloride were observed, and mapping analyses confirmed that these elements were actually contained within the corpora amylacea. A similar result was obtained using cryostat sections. Corpora amylacea are characteristically distributed along the margin of blood vessels, beneath the pial border of the hippocampus and in the subependymal zones of ventricles of aged brains, namely in the vicinity of blood and cerebrospinal fluid. From this distribution and from the results of the present paper, we suggest that corpora amylacea play a role in the absorption and accumulation of inorganic materials which have been extravasated from blood and cerebrospinal fluid (CSF) and taken up by astrocytes. This may reflect alteration of the blood-brain and blood-CSF barriers in the ageing brain.

Alzheimer Disease↗
PubMed1995

Molecular cloning, characterization, and chromosomal mapping of a novel human gene (GTF3A) that is highly homologous to Xenopus transcription factor IIIA.

We have isolated a novel human cDNA that is highly related to Xenopus transcription factor IIIA (TFIIIA). This clone contains an open reading frame of 1,269 nucleotides encoding 423 amino acids, including nine repeats of the Cys2His2-type of zinc-finger domain. A comparison of its sequence with Xenopus TFIIIA revealed 63% identity in nucleic acids and 58% identity in amino acids over a large portion of the gene and predicted peptide, indicating that the human homologue is likely to function as a transcription factor. The zinc-finger domains of the predicted protein also showed homology with those of human genes such as WT1, transcriptional repressor YY1, and MYC-associated zinc-finger protein (MAZ). Northern analysis showed expression in various tissues examined. The human TFIIIA gene (GTF3A) was localized to chromosome band 13q12.3-->q13.1 by fluorescent in situ hybridization (FISH).

Amino Acid Sequence↗
PubMed1995

Characterization of insulin-like growth factor-binding protein 5-degrading proteases produced throughout murine osteoblast differentiation.

Insulin-like growth factor (IGF)-binding protein-5 (IGFBP-5) is uniquely regulated throughout MC3T3-E1 osteoblast differentiation: IGFBP-5 is first detectable in conditioned medium (CM) of replicating preosteoblasts (day 5); IGFBP-5 levels peak between culture days 8-12, then decline to almost undetectable levels in mature osteoblast cultures (> day 18) despite the persistence of IGFBP-5 messenger RNA. These observations suggest that IGFBP-5 concentrations may be regulated by posttranslational mechanisms. To determine whether proteolysis contributes to the disappearance of IGFBP-5 in CM of mature osteoblasts, serial samples of MC3T3-E1 cell CM obtained during a 30-day culture period were analyzed for IGFBP-5-degrading protease activity. Using [125I]recombinant human IGFBP-5 substrate zymography, we demonstrated that proteases with M(r) of 52-72 and 97 kilodaltons (kDa) were present in CM, and protease activity increased in concentration as cultures matured. The 52- to 72-kDa proteases were cation dependent and were inhibited by tissue inhibitor of metalloproteinase 1, a specific inhibitor of matrix metalloproteinases (MMPs), identifying them as MMPs. Furthermore, antisera to human MMP-1 and -2 immunoprecipitated IGFBP-5-degrading proteases with M(r) of 52 and 69/72 kDa, respectively, suggesting that homologous murine MMPs degrade IGFBP-5. Finally, MC3T3-E1 cell CM contained immunoreactive MMP-1 and -2, and MMP-2, in particular, increased significantly throughout differentiation. In contrast, the 97-kDa protease was neither inhibited by tissue inhibitor of metalloproteinase 1 nor immunoprecipitated by antisera to MMPs, suggesting that the 97-kDa protease is not a MMP. Together, these data suggest that MMPs along with an unidentified 97-kDa protease degrade IGFBP-5 in MC3T3-E1 cell cultures. Because truncated forms of IGFBP-5 have been shown to enhance the action of IGF in bone cells, IGFBP-5 proteases may be instrumental in IGF-mediated bone morphogenesis.

Animals↗
PubMed1995

A 25.7 x 10(3) M(r) hydra metalloproteinase (HMP1), a member of the astacin family, localizes to the extracellular matrix of Hydra vulgaris in a head-specific manner and has a developmental function.

Hydra extracellular matrix (ECM) is composed of a number of components seen in vertebrate ECM such as laminin, type IV collagen, fibronectin, and heparan sulfate proteoglycan. A number of functional studies have shown that hydra ECM plays an important role in pattern formation and morphogenesis of this simple metazoan. The present study was designed to identify matrix degrading proteinases in hydra and determine their potential function in hydra morphogenesis. Using SDS-PAGE gelatin-zymography, five gelatinolytic bands were identified with relative molecular masses of 67 x 10(3), 51-58 x 10(3) (a triplet) and 25-29 x 10(3), respectively. Inhibition studies indicated that all of these gelatinases were metalloproteinases. Gelatin-zymography indicated that there was a differential distribution of these gelatinases along the longitudinal axis of hydra, with the 67 x 10(3) M(r) gelatinase being concentrated in the body column, while the 51-58 x 10(3) M(r) gelatinase triplet and the 25-29 x 10(3) M(r) gelatinase concentrated in the head region. Purification procedures were successfully developed for the 25-29 x 10(3) M(r) metalloproteinase which has been termed hydra metalloproteinase 1 (HMP1) and which appeared as a single band with a SDS-PAGE mobility of 25.7 x 10(3) M(r). The N-terminal sequence of purified HMP1 indicated that it has structural homology with metalloproteinases that belong to the astacin family. Subsequent cloning and sequencing of cDNA clones confirmed the identification of HMP1 as an astacin-like metalloproteinase. Immunocytochemical studies with antibodies generated against the purified enzyme and to a synthetic peptide indicated that HMP1 was localized to the ECM of tentacles. Functional studies were performed in which purified HMP1, anti-HMP1 IgG, or suspected substrates of HMP1 (e.g. growth factors such as TGF-beta 1) were introduced into the interepithelial compartment of hydra using a 'DMSO loading' procedure. These studies indicated that HMP1 has a functional role during a number of developmental processes such as head regeneration and cell differentiation/transdifferentiation of tentacle battery cells.

Amino Acid Sequence↗
PubMed1995

Matrix metalloproteinases and their tissue inhibitors at the ovine trophoblast-uterine interface.

Trophoblast invasiveness in ruminants is limited to fusion of migrating binucleate cells with uterine epithelium, but considerable tissue remodelling and angiogenesis occurs within the endometrium at implantation. Such processes are elsewhere associated with an altered balance of matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). ProMMPs-1, -2, -3 are secreted by cultured ovine endometrial stromal, but not epithelial, cells and expression of proMMP-1 and proMMP-3 is inhibited by interferon tau, the dominant preimplantation trophoblast product, independently of its effects on prostaglandin production. Messenger RNA for TIMP-1 and TIMP-2 is expressed in endometrium throughout the oestrous cycle and early pregnancy. Both TIMP-1 mRNA (0.9 kb transcript) and TIMP-2 mRNA (1.0 kb transcript) expression increase at day 12 through to day 20, suggesting a role in preventing trophoblast invasion. Expression of the 3.5 kb transcript for TIMP-2 decreases from day 16 and is undetectable on day 20. Uterine flushings from both nonpregnant and pregnant ewes on day 16 after oestrus contain proMMP-2, possibly transudated from plasma, and proMMP-9, but concentrations are higher in pregnancy. These enzymes are also detected in conditioned medium following culture of trophoblasts at days 16-20. Whether the trophoblast MMPs have a role in implantation in sheep remains to be established. The production of MMPs and TIMPs in the endometrium and the changes associated with implantation events suggest that they contribute to the marked edometrial remodelling associated with early placentation.

Animals↗
PubMed1995

Modification of mu-opioid agonist-induced locomotor activity and development of morphine dependence by diabetes.

We examined the locomotor-enhancing action of mu-opioid receptor agonists, such as morphine and [D-Ala2, N-MePhe4, Gly-ol5]enkephalin (DAMGO), and physical dependence on morphine in diabetic and nondiabetic mice. Morphine (5-20 mg/kg, s.c.) and DAMGO (1-4 nmol, i.c.v.) had a dose-dependent locomotor-enhancing effect in both nondiabetic and diabetic mice. The locomotor-enhancing effects of morphine and DAMGO were significantly less in diabetic mice than in nondiabetic mice, and were significantly reduced after pretreatment with either beta-funaltrexamine (20 mg/kg, s.c.), a selective mu-opioid receptor antagonist, or naloxonazine (35 mg/kg, s.c.), a selective mu1-opioid receptor antagonist. Both diabetic and nondiabetic mice were chronically treated with morphine (8-45 mg/kg, s.c.) for 5 days. During this treatment, neither diabetic nor nondiabetic mice showed any signs of toxicity. After morphine treatment, withdrawal was precipitated by injection of naloxone (0.3-10 mg/kg, s.c.). Several withdrawal signs, such as weight loss, diarrhea, ptosis, jumping and body shakes, were observed after naloxone challenge in morphine-dependent nondiabetic mice. Although morphine-dependent diabetic mice showed greater weight loss than nondiabetic mice, the incidence of jumping and body shakes after naloxone challenge in diabetic mice were lower than that in nondiabetic mice. These results suggest that diabetic mice are selectively hyporesponsive to mu1-opioid receptor-mediated locomotor enhancement. Furthermore, diabetes may affect mu1-opioid receptor-mediated naloxone-precipitated signs of withdrawal from physical dependence on morphine.

Animals↗
PubMed1995

[Endocrine mechanism of placental circulatory disturbances induced by microwave in pregnant rats].

Effects of microwaves on fetus and female genital organs remain to be elucidated. To demonstrate the placental circulatory disturbances induced by microwaves and to clarify the endocrine pathogenesis, placental blood flow and five endocrine indicators, i.e., corticosterone (CS), estradiol (E2), progesterone (P), prostaglandin E2 (PGE2) and prostaglandin F2 alpha (PGF2 alpha) were measured in rats exposed to whole-body microwaves with an intensity of 10 mW/cm2 at a frequency of 2,450 MHz. The placental blood flow at 45-90 min after exposure was significantly decreased in the rats exposed to the microwaves. Placental blood flow at 15 and 30 min was increased by pretreatment with intraperitoneal administration of angiotensin II (AII). In contrast, no significant change in placental blood flow was recognized in the AII pretreated rats exposed to the microwaves. An increase in CS and a decrease in E2 were induced by the microwave exposure independent of pretreatment with AII. P was increased by microwave exposure in the rats without pretreatment with AII. PGE2 was not changed by the microwave exposure in the case of either nonpretreatment or pretreatment with AII. PGF2 alpha was increased by the microwave exposure in the rats without pretreatment with AII. The present results indicate that excessive exposure to whole-body microwave disorders pregnancy in terms of placental circulatory dysfunction. The data suggest the involvement of endocrine mechanisms in the decrease in placental blood flow which is induced via a detrimental effect of microwaves on PGF2 alpha and on pituitary functions such as general emotional stress.

Angiotensin II↗
PubMed1995

Effect of diabetes on the morphine-induced inhibition of gastrointestinal transit.

The effect of diabetes on the morphine-induced inhibition of gastrointestinal transit was examined in mice. Morphine dose-dependently inhibited gastrointestinal transit after s.c. administration in both non-diabetic mice and diabetic mice. There was no significant difference between the ED50 values for this antitransit effect of morphine in non-diabetic and diabetic mice. The gastrointestinal antitransit effect of morphine was significantly antagonized by pretreatment with beta-funaltrexamine (40 mg/kg, s.c.), a selective mu-opioid receptor antagonist, in both non-diabetic and diabetic mice. However, pretreatment with naloxonazine (35 mg/kg, s.c.), a selective mu 1-opioid receptor antagonist, had no effect on the antitransit properties of morphine. These results suggest that diabetes failed to alter the mu 2-opioid receptor-mediated antitransit effect of morphine.

Animals↗
PubMed1995

Involvement of endothelin in peripheral circulatory change induced by hand-arm vibration.

Physiologic mechanisms involving local vasoregulating factor, endothelium-derived relaxing factor (EDRF) and endothelium-derived constricting factors (EDCF) have been postulated to play a role in VWF. Recent evidence that endothelin is a potent vasoconstrictor peptide indicates that it may play a role in vasoregulation during vibration exposure through the local actions of EDRF or EDCF. Therefore, we examined the effects of grasping (50 N) and hand-arm vibration with an unweighted acceleration of 50 m/s2 rms at a frequency of 120 Hz in the direction of X-axis on digital blood flow (DBF) and on the level of plasma endothelin in 7 healthy male office workers. DBF was measured by an apparatus based on the thermal diffusion method where the thermal diffusion flow probe was incorporated in the Peltier's stack. In the grasping test, DBF decreased by about 40% from resting level within 1 minute after the grasping was begun, and was maintained at almost the same level during grasping, but increased immediately after cessation of grasping. In the vibration exposure test, initial decrease in DBF due to grasping was the same as that observed in the grasping test, but DBF gradually increased when exposure to vibration was performed simultaneously. No significant change in plasma endothelin level was induced by grasping. Endothelin was significantly lower after exposure to vibration than at rest and after grasping. The grasping-induced decrease in DBF seemed to be due to mechanical compression of the vessels. The negative correlation between DBF and endothelin during vibration exposure suggests that a reduction in release of endothelin from smooth muscle into the vessel cavity during vibration leads to vasodilatation, possibly attributable to the local axon reflex.

Blood Flow Velocity↗
PubMed1995

Importance of matrix metalloproteinases in human trophoblast invasion.

Human cytotrophoblast cells are invasive by virtue of their ability to secrete metalloproteinases (MMP) capable of digesting the extracellular matrix of the endometrium. It is the aim of the present study to determine which of the known MMP is responsible for this invasive behavior and to see to what extent endometrial secretions can modulate this enzymatic activity. Under our experimental conditions, first-trimester cytotrophoblast cells invade matrigel; this invasive behavior is inhibited by phenanthroline (an inhibitor of MMP) and by a polyclonal antibody to the 92-kDa gelatinase but not to other MMP. Since cytotrophoblast cells cultured in vitro secrete the 92-kDa gelatinase, and since adhesion to a substrate increases their gelatinolytic activity, it is believed that cytotrophoblast cells invade their surrounding matrix by binding to it and by increasing their secretion of 92-kDa gelatinase which then digests the collagen type IV of their micro-environment. This process of invasion is controlled by secretions from decidual cells (but not from non-decidualized stromal cells) since conditioned medium from decidual cells inhibits the activity of the 92-kDa gelatinase released from cytotrophoblast cells.

Animals↗
PubMed1994

Matrix metalloproteinases degrade insulin-like growth factor-binding protein-3 in dermal fibroblast cultures.

Insulin-like growth factor binding protein-3 (IG-FBP-3) is degraded by a Zn(2+)-dependent protease(s) produced by human dermal fibroblasts in vitro (Fowlkes, J. (1994) Endocrine J. 2, 63-68). Initial studies using IG-FBP-3-substrate zymography identified several IGFBP-3-degrading proteases with M(r) 52,000-72,000, which were inhibitable by EDTA and were shifted to lower M(r) species after treatment of conditioned medium with an organomercurial, suggesting that they might represent one or more of the matrix metalloproteinases (MMPs). Immunoblotting of conditioned medium demonstrated the presence of proMMP-1 (52 and 55 kDa), proMMP-3 (58 and 60 kDa), and proMMP-2 (72 kDa) whose molecular masses corresponded identically to those of the IGFBP-3-degrading proteases. Degradation of recombinant human (rh) IGFBP-3 by conditioned media was blocked (> 80% inhibition) by tissue inhibitor of metallo-proteinases-1, a specific inhibitor of all MMPs, while removal of MMPs -1, -2, and -3 from conditioned medium by sequential immunoaffinity and gelatin-Sepharose chromatography resulted in the complete loss of IGFBP-3-degrading proteinase activity. Furthermore, human MMP-1, MMP-3, and to a lesser extent MMP-2 degraded rhIGFBP-3 in vitro. Sequence analysis of rhIGFBP-3 cleavage sites produced by MMP-1, -2, or -3 demonstrated that each cleaved within the mid-region of the binding protein, a domain with little or no homology with the other five cloned IGFBPs. These studies suggest that MMPs, beyond their previously described functions as extracellular degrading enzymes, may also exert effects on cellular growth and proliferation via degradation of IGFBP-3, thus enhancing IGF bioavailability.

Amino Acid Sequence↗