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T Terao

Publications and source records attributed to T Terao.

At least 199 records · Page 11Linked to original sources
PubMed1996

Biochemical changes in the cervical mucus after application of laminaria tent.

BACKGROUND: This research was aimed to study the biochemical changes in cervical mucus after application of laminaria tent. MATERIAL AND METHODS: Cervical mucus was collected before and after insertion of laminaria tent from 20 normal pregnant women in the first and second trimesters. Interleukin-1 beta, IL-8, collagenase activity, elastase activity, PGE2 and PGF2 alpha were measured in the collected samples. Laminaria tent was also inserted in one vaginal cervix of five non-pregnant rabbits and the clinical and histological changes were compared with five non-treated rabbits. Collagen contents were assessed histologically and Leukocyte number was counted. RESULTS: IL-1 beta and IL-8 and elastase activity were significantly increased after insertion of laminaria (p<0.0001 and 0.001 respectively). Collagenase activity was not significantly changed. Whereas concentration of PGE2 and PGF2 alpha in the mucus were markedly increased (p<0.0001 and 0.001 respectively). Collagen contents were significantly decreased in laminaria and opposite side cervices (p<0.0001). No significant changes in leukocyte count could be observed in treated and non treated cervices. CONCLUSIONS: Insertion of laminaria leads to increase concentration of IL-1 beta, IL-8, PGE2, PGF2 alpha and elastase activity. Mechanical stretching of the cervix and biochemical changes in cervical mucus may explain the mechanism of cervical ripening induced by laminaria.

Animals↗
PubMed1996

Expression of interleukin-8 receptors in the gestational tissues before and after initiation of labor: immunohistochemical study.

BACKGROUND: Interleukin-8 (IL-8) has an essential role in the mechanism of parturition. IL-8 induces its effect through specific cell binding receptors. The main purpose of this research was to elucidate the presence of IL-8 receptors in the gestational tissues and their relation to parturition. MATERIALS AND METHODS: Fetal membranes, placenta, umbilical cord and myometrial samples were collected before the initiation of labor during elective Cesarean sections (n = 15). Similar biopsies (except myometrial samples) were also gathered after normal vaginal delivery (n = 15). Immunohistochemical staining for IL-8 receptors type I and II was carried out for the fresh frozen sections of all samples. Immunoreactive score (IRS) was calculated for the stained sections. RESULTS: IL-8 Receptors type I and II were recognized in the fetal membranes, placenta, umbilical cord and myometrial samples collected from patients before the initiation of labor. The staining of amnion and placentas were found to be more intensified in the samples collected after vaginal delivery compared to those from elective cesarean sections and before initiation of labor. CONCLUSIONS: Increased expression of IL-8 receptors after initiation of labor may indicate that IL-8 receptors might have a role during parturition.

Antigens, CD↗
PubMed1996

[Comparison of biochemical properties of human airway tryptase isolated from mucoid sputum with those of lung mast cell tryptase].

We found a novel trypsin-like enzyme (tryptase) in sputum from patients with chronic airway diseases, and named this enzyme human airway tryptase (HAT). To clarify its physiological significance in the airway, we compared biochemical properties of purified HAT with those of purified lung mast cell tryptase (MCT). Studies with model peptide substrates showed that both the HAT and MCT preferentially cleaved the COOH-terminal side of arginine residues of certain peptides, but substrate specificities to nine synthetic model substrates of HAT differed from those of MCT. Effects of protease inhibitors on the two enzymes were examined at a concentration of 10 microM. Both the HAT and MCT were strongly inhibited by the trypsin inhibitors leupeptin, antipain, and aprotinin. An alpha-1-protease inhibitor inhibited HAT by 50%, but it did not inhibit MCT. In contrast, a secretory leukocyte protease inhibitor strongly inhibited MCT, but not HAT. Mucoid sputum from patients with chronic bronchitis contained much more HAT than MCT. These differences in biochemical properties between HAT and MCT indicate that they play different physiological roles in the airways.

Asthma↗
PubMed1996

Induction of HELLP syndrome-like biochemical parameters by stimulation of the celiac ganglion in rats.

OBJECTIVE: An animal model of HELLP (hemolysis, elevated liver enzymes, low platelets) syndrome was developed by means of stimulation of the celiac ganglion in rats. METHODS: The celiac ganglion in pregnant or non-pregnant rats was exposed to endotoxin (lipopolysaccharide, LPS) (500 micrograms/50 microliters), potassium chloride (0.2 mol/l/50 microliters), or saline solution (50 microliters). In another group of rats the bifurcation of the abdominal aorta was exposed to LPS (500 micrograms/50 microliters). Blood pressure, platelet count, hematocrit, serum aspartate transaminase (AST), alanine transaminase (ALT), lactate dehydrogenase (LDH), and plasma norepinephrine and epinephrine were measured for 6 h after treatment. Histopathologic studies were also performed in these rats. RESULTS: A significant increase in blood pressure, AST, ALT, LDH, norepinephrine, and epinephrine was found in the endotoxin-treated pregnant rats compared with control rats treated with the saline solution. A significant decrease in platelet count was found in endotoxin-treated pregnant rats compared with the control rats. A significant increase in blood pressure, AST, norepinephrine, and epinephrine was found in the potassium chloride-treated pregnant rats compared with control rats. Blood pressure and biochemical parameters remained unchanged in the pregnant rats treated with LPS at the bifurcation of the abdominal aorta, as in those treated with saline at the celiac ganglion. Histologic examination of liver tissues treated with LPS or potassium chloride showed varying degrees of ischemic necrosis of hepatocytes similar to that observed in the human HELLP syndrome. Blood pressure, biochemical parameters, and histologic findings in non-pregnant rats were almost the same as those in pregnant rats. CONCLUSION: This study suggests that exogenous stimulation of the celiac ganglion causes an increase in the blood pressure and liver ischemia, resulting in HELLP syndrome-like disease in pregnant and non-pregnant rats.

Animals↗
PubMed1996

Urinary trypsin inhibitor efficiently inhibits urokinase production in tumor necrosis factor-stimulated cells.

We demonstrated that urinary trypsin inhibitor (UTI) efficiently inhibits soluble and tumor cell-associated plasmin activity and subsequently inhibits tumor cell invasion and metastasis. The effect of UTI on tumor necrosis factor-alpha (TNF)-induced stimulation of urokinase-type plasminogen activator (uPA) in cultured human umbilical vein endothelial cells (HUVEC) and in the promyeloid leukemia U937 cells was studied. uPA antigen was evaluated in the cell lysate and in the conditioned media by enzyme-linked immunosorbent assay, sodium dodecyl sulfate polyacrylamide gel electrophoresis, and Western blot. TNF can promote the production of uPA in HUVEC and in U937 cells. The PKC inhibitors (H7, calphostin C, and staurosporine) inhibited TNF-induced uPA expression and secretion in a dose-dependent manner. Analysis of the expression of cell surface receptor-bound uPA by flow cytometry using uPA-specific MAb indicates that induction of uPA expression by TNF was inhibited when these cells were incubated with UTI. On the other hand, treatment of the cells with UTI alone failed to alter uPA production. UTI also reduced the secretion of uPA in TNF-treated cells. UTI was as effective as PKC inhibitors in inhibiting uPA expression by TNF. Incubation of the cells with UTI, however, had no effect on the ability of PMA to stimulate cell-associated uPA expression. These data suggest that UTI may influence the PKC-dependent protein kinase pathway in uPA expression. The study on intracellular pathways involved in UTI modulation of uPA will enhance our understanding of the role that UTI plays in uPA-mediated cellular invasion.

8-Bromo Cyclic Adenosine Monophosphate↗
PubMed1996

Biochemical analysis of airway aspirates of newborns.

We measured contents of tracheobronchial and alveolar components contained in the newborn airway aspirates (NAA) obtained from 54 healthy newborns, with bronchoalveolar lavage fluid (BALF) and bronchial lavage fluid (BLF) from healthy adults, and with mucoid sputum from adults with chronic bronchitis as controls. Fucose (a parameter of tracheobronchial mucus mucoprotein) and tryptase activity were used as tracheobronchial components, and pulmonary surfactant apoprotein A (SP-A) as an alveolar component. The ratio of the content of each component to that of total protein (TP) was compared among NAA, BLF, BALF, and the mucoid sputum. The SP-A/TP ratio in the NAA was similar to that in BLF but 1/24 of that in BALF. The fucose/TP ratio in NAA was about 1/2 of that in BLF and about 6 times higher than that in BALF. The tryptase/TP ratio in NAA was about twice that in BLF and about 80 times that in BALF. The ratios of the above components to TP in the mucoid sputum were closer to those in NAA and BLF than those in BALF. These results suggest that NAA contain both tracheobronchial and alveolar components, and their contents of the formers are larger than those of the latters, and that NAA may be useful for diagnosing pathologic conditions in the airway of newborns, and for comparing the biological defence mechanism between newborns and adults.

Adult↗
PubMed1995

Inhibition of metastasis of Lewis lung carcinoma by urinary trypsin inhibitor in experimental and spontaneous metastasis models.

A purified human urinary trypsin inhibitor (UTI) and its related synthetic peptides were examined to determine whether they could inhibit production of experimental and spontaneous lung metastases by murine Lewis lung carcinoma (3LL) cells. Three peptides, peptide I, peptide 2 and peptide 3, representing the amino acid sequences within the UTI molecule, were synthesized. UTI and peptide 2 inhibited human leukocyte elastase (HLE). UTI and peptide 3 specifically inhibited human and murine plasmin activity. Peptide I had essentially no inhibitory activity. In an in vivo spontaneous metastasis model, multiple s.c. injections of UTI or peptide 3 for 7 days immediately after s.c. tumor cell inoculation significantly inhibited the formation of lung metastasis in C57BL/6 mice in a dose-dependent manner. UTI reduced lung tumor colonization more effectively than peptide 3. Peptides 1 and 2, however, did not affect the formation of lung metastasis. Inhibition of lung metastasis was not due to direct anti-tumor effects of UTI and peptide 3. In an in vivo experimental metastasis assay, multiple s.c. injections of UTI for 7 days after i.v. tumor cell inoculation inhibited metastatic lung tumor colonization, while peptide 3 did not affect metastasis. Peptides 1 and 2 did not affect the formation of lung metastasis. When examined with an in vitro assay system using a modified Boyden chamber, UTI and peptide 3 suppressed the invasion of tumor cells through Matrigel. UTI and peptide 3 inhibited neither cell proliferation nor the binding of tumor cells to Matrigel and showed no significant suppression of chemotactic migration of tumor cells to fibronectin. Our results suggest that UTI efficiently regulates the mechanism involved in not only the entry into vascular circulation of tumor cells (intravasation, though, at least in part, inhibition of the proteolytic enzyme plasmin) but also the extravasation step of the metastatic process.

Amino Acid Sequence↗
PubMed1995

Endothelin-1 evoked an increase and oscillations in cytosolic calcium concentration in adherent single human platelets and increased GMP-140 (P-selectin) in platelet suspension.

The effect of endothelin-1 (ET-1) on cytosolic calcium ion concentration ([Ca2+]i) in adherent single human blood platelet was determined by fluorescence digital imaging microscopy using Fura-2 as calcium probe. A dose dependent increase and oscillatory changes in [Ca2+]i in single platelets were evoked by ET-1 as with thrombin. Half and 1 microM of ET-1 increased (p < 0.01) the [Ca2+]i in single platelets from a resting level of 83 +/-3.4 nM to 120 +/- 13 nM and 240 +/- 20 nM respectively. The ET-1 induced increase in [Ca2+]i was suppressed by 1 mM EGTA, a calcium chelating agent in the medium. ET-1 increased the production of IP3 (quantitified by IP3 3H-radioreceptor assay kit) in platelets significantly (p < 0.05) in a dose dependent way. We measured the GMP-140 (P-selectin) level in the supernatant of human platelet suspensions incubated with thrombin and ET-1. Both thrombin and ET-1 increased the secretion of soluble GMP-140 in the supernatant of platelet suspensions. Therefore, we suggested that ET-1 increased [Ca2+]i in platelets by both calcium influx and IP3 mediated Ca2+ release resulting in activation and release of GMP-140.

Adult↗
PubMed1995

Up-regulation of dystrophin mRNA by exposure to dibutyryl cAMP in the C2C12 muscle cell line.

The effect of dibutyryl cAMP on dystrophin mRNA expression was investigated in C2C12 myogenic cells by a semi-quantitative RT-PCR method. The dystrophin mRNA level was enhanced by the addition of low concentrations of dibutyryl cAMP for 16h in the growth medium. The relative amounts of dystrophin/G3PDH mRNA at 10(-4) M dibutyryl cAMP were about 2.6-fold higher compared to control. We examined MLC1a mRNA expression in order to know whether the induction of dystrophin mRNA is due to the facilitation of myogenic differentiation by dibutyryl cAMP. MLC1a mRNA expression at 10(-4) M dibutyryl cAMP was similar to that of control. Thus we conclude that the dibutyryl cAMP enhancement of dystrophin mRNA expression is not due to a secondary event involving the formation of differentiated myotubes. The induction of CREB mRNA by dibutyryl cAMP suggests that the up-regulation of dystrophin mRNA expression might occur via transcriptional activation.

Animals↗
PubMed1995

Inhibition of tumor cell invasion through matrigel by a peptide derived from the domain II region in urinary trypsin inhibition.

Urinary trypsin inhibitor (UTI) has a multipotent inhibitory effect on proteases such as trypsin, chymotrypsin, plasmin, human leukocyte elastase, or hyaluronidase. UTI can bind easily to its receptors on various types of tumor cells (human ovarian cancer HOC-I cells, human choriocarcinoma SMT-cc1 cells, and murine Lewis lung carcinoma 3LL cells). Our results show that the UTI receptors of some tumor cells have a possible role in modulating plasmin activity on the cell surface and prevention of tumor cell invasion and metastasis (H. Kobayashi et al., J. Biol. Chem., 269; 20642-20647, 1994). UTI interacts with tumor cells as a negative modulator of the invasive cells. We investigated whether this effect may be mediated by UTI binding to the cell surface receptors. In addition, the role of peptide sequences from each UTI domain and their interaction with tumor cells were investigated. UTI derivatized with biotin or FITC was taken up by tumor cells in a dose-dependent manner. This cell association was inhibited with a monoclonal antibody D1, which specifically recognizes NH2 terminus (domain I) of UTI. The binding was inhibited by fluid phase UTI, but not HI-8, COOH terminus (domain II) of UTI, suggesting that UTI binds to cells through a site in the UTI domain I. Furthermore, we found that UTI, HI-8 and a number of peptides containing Arg-Gly-Pro-Cys-Arg-Ala-Phe-Ile promoted the inhibition of tumor cell invasion. This site corresponds to the plasmin-inhibiting domain within HI-8. The possibility that UTI binding to tumor cells might be involved in the prevention of tumor cell invasion in vitro was excluded since HI-8, lacking domain I, promotes the inhibition of tumor cell invasion with essentially the same affinity as UTI. All these data allow us to conclude that inhibition of tumor cell invasion is mediated by domain II, which possesses anti-plasmin activity.

Amino Acid Sequence↗
PubMed1995

Inhibitory effect of a conjugate between human urokinase and urinary trypsin inhibitor on tumor cell invasion in vitro.

Proteolytic enzymes such as urokinase-type plasminogen activator (uPA), plasmin, and collagenase mediate proteolysis by a variety of tumor cells. uPA secreted by tumor cells can be bound to a cell surface receptor via a growth factor-like domain within the amino-terminal fragment (ATF) of the uPA molecule with high affinity. Urinary trypsin inhibitor (UTI) efficiently inhibits the soluble and the tumor cell-surface receptor-bound plasmin and subsequently reduces tumor cell invasion and the formation of metastasis. The anti-invasive effect is dependent on the anti-plasmin activity of the UTI molecule, domain II in particular. We synthesized a conjugate between ATF of human uPA and a native UTI molecule or domain II of UTI (HI-8). The effect of the conjugates (ATF.UTI or ATF.HI-8) on tumor cell invasion in vitro was investigated. ATF.UTI and ATF.HI-8 bound to U937 cells in a rapid, saturable, dose-dependent, and reversible manner. A large part of receptor-bound ATF-UTI and ATF.HI-8 remains on the cell surface for at least 5 h at 37 degrees C. Inhibition of tumor cell-surface receptor-bound plasmin by ATF.UTI and ATF.HI-8 was markedly enhanced when compared with tumor cells treated either with ATF, UTI, or HI-8. Results of a cell invasion assay showed that ATF.UTI and ATF.HI-8 is very effective at targeting HI-8 specifically to uPA receptor-expressing tumor cells, whereas tumor cells devoid of uPA receptor may be less affected by the conjugates. Our results indicate that cell surface uPA and plasmin activity is essential to the invasive process and that the conjugates exhibit plasmin inhibition to the close environment of the cell surface and subsequently inhibit the tumor cell invasion through Matrigel in an in vitro invasion assay.

Amino Acid Sequence↗
PubMed1995

Activated neutrophil by endothelin-1 caused tissue damage in human umbilical cord.

Immunostaining of human neutrophils incubated with endothelin-1 (ET-1) showed intense and spreading pattern of anti human granulocyte elastase within the cytosol. That reflected neutrophil activation followed by the release of granule contents by ET-1. In contrast, PBS (phosphate buffered saline) treated neutrophils showed localized and faintly stained granules. Intracellular calcium in fura-2 loaded neutrophils was measured at 340/380 nm. A dose and time dependent increase in intracellular calcium by ET-1 occurred in human single neutrophil. Elastase activity assay was done with chromogenic substrate S2484. ET-1 induces dose and time dependent increase in elastase activity in neutrophil suspensions like ionophore A23187. A similar time dependent increase in elastase activity was retained even after repeated wash and ET-1 treatment. That confirmed the viability of most of the neutrophils after each treatment. In umbilical cord preparations, ET-1 treated neutrophils could migrate from the venous lumen into the tissue matrix of the umbilical cords. Hematoxylin and eosin staining revealed a massive tissue destruction in ET-1 activated neutrophil treated cords when compared to sham control and untreated neutrophil injected cords. Immunostaining with monoclonal anti human elastase revealed an intense staining in former sections when compared to the others. We suggest that ET-1 activated neutrophil might play a major role in endothelial injury and tissue damage in conditions with high blood level of endothelin.

Calcium↗
PubMed1995

Kunitz-type trypsin inhibitor prevents LPS-induced increase of cytosolic free Ca2+ in human neutrophils and HUVEC cells.

The protease inhibitor part of inter-alpha trypsin inhibitor is identical to urinary trypsin inhibitor (UTI). Preincubation of neutrophils and HUVEC cells with UTI inhibited increase of cytosolic free Ca2+ induced by LPS. Increase of cytosolic free Ca2+ induced by LPS in the presence of EGTA was also inhibited by UTI. In contrast, UTI did not inhibit increase of cytosolic free Ca2+ in cells stimulated by Ca2+ ionophore with or without EGTA. The effects of nine synthetic peptides of UTI on the concentration of cytosolic free Ca2+ in the neutrophils induced by LPS were examined. Preincubation with a peptide of UTI domain 2, NLPIVRGPCRAFIQL (83-97), was completely inhibited by the increase of cytosolic free Ca2+ in neutrophils. This region is identical to the trypsin inhibitor site of UTI. We propose that a function of UTI other than as a protease inhibitor is in regulation of intracellular Ca2+ and that this is due to its trypsin inhibitor region.

Amino Acid Sequence↗
PubMed1995

Down-regulation of interleukin-8 gene expression in HL60 cell line by human Kunitz-type trypsin inhibitor.

Urinary trypsin inhibitor (UTI) is one of the Kunitz-type protease inhibitors in human. Little is known about its anti-inflammatory functions other than protease inhibition. We studied the effect of UTI on gene expression of interleukin-8 (IL-8), an inflammatory cytokine. UTI inhibited IL-8 gene expression induced by lipopolysaccharide (LPS) in HL60 cells. The IL-8 concentrations in the cells and medium after LPS stimulation increased time-dependently in the absence of UTI, but did not increase in the presence of UTI. On the other hand, UTI did not inhibit either the synthesis or the release of IL-8 induced by the calcium ionophore A23187. UTI inhibited increase of cytosolic Ca2+ stimulated by LPS but not by A23187. Our results suggest that the inhibition by UTI is due to its effect on the cell membrane involved in regulating Ca2+ influx.

Calcimycin↗