Fibroblast growth factor-4 (FGF-4) has various functions, affecting many signaling pathways, and leading to cellular proliferation and differentiation and to the regulation of cell migration, invasion, and angiogenesis. However, there are few reports of the relationship between TS cells and FGF-4 even if FGF-4 is located in inner cell mass of embryo and Fibroblast growth factor receptor (FGFR) is located in TS cells. Therefore the physiologic effects of FGF-4 on TS cells were investigated for identifying the effects of FGF-4 on TS ell differentiation. FGF-4 was involved in early stage development of the trophoblast via upregulation of eomesodermin mRNA expression. In addition, FGF-4 suppressed the differentiation of TS cells through activation of extracellular-signal regulated kinase (Erk) and suppression of focal adhesion kinase (FAK) activation, which in TS cells is an important indicator of early trophoblast cell differentiation, migration and invasion. FGF-4 was involved in angiogenesis in the trophoblast through the activation of p38 and the induction of Dlx-3 mRNA expression in TS cells. In addition, TS cells cultured with FGF-4 for 4 days in a thrombinfibrinogen gel culture system, a specific culture system for endothelial cells, showed a healthy appearance, while TS cells cultured without FGF-4 were severely damaged. Taken together, these data suggest that FGF-4 is closely involved in differentiation of TS cells for development of placenta.
Cell Differentiation ; Cell Movement ; Cell Proliferation ; Embryonic Structures ; Endothelial Cells ; Fibroblasts ; Focal Adhesion Protein-Tyrosine Kinases ; Phosphotransferases ; Placenta ; Receptors, Fibroblast Growth Factor ; RNA, Messenger ; Trophoblasts* ; Up-Regulation

BACKGROUND/AIMS: Ischemic colitis is a vascular condition of inadequate blood flow in the colon which leads to colonic inflammation and can cause significant morbidity and mortality. Oxidative stress is an early initiating event in ischemia and reperfusion injury. Heme oxygenase (HO) is considered to be an antioxidant enzyme that catabolizes heme to carbon monoxide, free iron and biliverdin. The aim of this study was to evaluate the expression patterns of HO-1, inducible form of HO, in ischemic colitis. METHODS: We analyzed the twelve cases of clinically and pathologically diagnosed ischemic colitis without surgical intervention compared with normal colon (n=10) and psedomembranous colitis (n=5). Immunohistochemical stainings for HO-1 were performed in paraffin-embedded tissues. RESULTS: The age of the patients ranged from 56 to 84 years (mean: 67 years) in ischemic colitis. Eight patients (66.7%) were female. The most common presenting symptom was bloody stool (66.7%) and rectosigmoid area (91.7%) of the large intestine was the most common ischemic site. Expression of HO-1 in ischemic colitis was high in contrast to normal colonic mucosa or psedomembranous colitis. CONCLUSIONS: Ischemic colitis usually involves the rectosigmoid area in elderly female patients with a history of bloody stool. High expression of HO-1 in ischemic colitis may be responsible for a protective mechanism to ischemia or heme injury.
Aged ; Aged, 80 and over ; Colitis, Ischemic/*enzymology ; Colon/*enzymology ; Female ; Heme Oxygenase (Decyclizing)/*metabolism ; Humans ; Immunohistochemistry ; Intestinal Mucosa/*enzymology ; Male ; Middle Aged

BACKGROUND: The serotonin type 3 receptors are diffusely distributed in both the central and the peripheral nervous system. Physiological and pathophysiological processes thought to be mediated by this receptor include nausea and vomiting, peripheral nociception and central antinociception, conditioned aversion response to drugs, anxiety, and cognition. Because of the structural similarity between the nicotinic acetylcholine receptor and the 5HT3 receptor, we investigated the effects of clinically used neuromuscular blockers on the 5HT3 receptor function related with PONV. METHODS: A cDNA clone encoding the full length murine 5HT3a receptor was subcloned into an oocyte expression vector and 50 ng of cRNA transcribed in vitro injected per oocyte. After 24 72 h incubation, oocytes were placed into a recording chamber continuously perfused with frog Ringer's solution and electrophysiological recordings were obtained by the two electrode voltage clamp technique. Serotonin with or without the various drugs were bath applied by a computer controlled solenoid valve. Peak currents induced by the drug applications were measured and dose responses were obtained. RESULTS: The 5HT3 receptor expression in Xenopus oocyte was identified by the pharmacologic tools. Serotonin induced rapid inward currents, and thus was showed dose-dependent: KD = 2.5 micrometer, Hill coefficiency = 2.09. Inhibition by the neuromuscular blockers showed dose-dependence and their inhibitory potency on 5HT3 receptor (IC50) was in order of d-tubocurarine (0.046 micrometer) > vecuronium (16.32 micrometer) > gallamine (1,169 micrometer). CONCLUSIONS: There was a different inhibitory effect of nicotinic cholinergic antagonists, clinically used neuromuscular blockers, on the 5HT3 receptor and a judicious selection of them might contribute to reducing the incidence of PONV clinically.
Anxiety ; Baths ; Cholinergic Antagonists ; Clone Cells ; Cognition ; DNA, Complementary ; Electrodes ; Gallamine Triethiodide ; Incidence ; Nausea ; Neuromuscular Blockade* ; Neuromuscular Blocking Agents* ; Nociception ; Oocytes* ; Peripheral Nervous System ; Postoperative Nausea and Vomiting ; Receptors, Nicotinic ; RNA, Complementary ; Serotonin* ; Tubocurarine ; Vecuronium Bromide ; Vomiting ; Xenopus*

BACKGROUND: The serotonin type 3 receptors are diffusely distributed in both the central and the peripheral nervous system. Physiological and pathophysiological processes thought to be mediated by this receptor include nausea and vomiting, peripheral nociception and central antinociception, conditioned aversion response to drugs, anxiety, and cognition. Because of the structural similarity between the nicotinic acetylcholine receptor and the 5HT3 receptor, we investigated the effects of clinically used neuromuscular blockers on the 5HT3 receptor function related with PONV. METHODS: A cDNA clone encoding the full length murine 5HT3a receptor was subcloned into an oocyte expression vector and 50 ng of cRNA transcribed in vitro injected per oocyte. After 24 72 h incubation, oocytes were placed into a recording chamber continuously perfused with frog Ringer's solution and electrophysiological recordings were obtained by the two electrode voltage clamp technique. Serotonin with or without the various drugs were bath applied by a computer controlled solenoid valve. Peak currents induced by the drug applications were measured and dose responses were obtained. RESULTS: The 5HT3 receptor expression in Xenopus oocyte was identified by the pharmacologic tools. Serotonin induced rapid inward currents, and thus was showed dose-dependent: KD = 2.5 micrometer, Hill coefficiency = 2.09. Inhibition by the neuromuscular blockers showed dose-dependence and their inhibitory potency on 5HT3 receptor (IC50) was in order of d-tubocurarine (0.046 micrometer) > vecuronium (16.32 micrometer) > gallamine (1,169 micrometer). CONCLUSIONS: There was a different inhibitory effect of nicotinic cholinergic antagonists, clinically used neuromuscular blockers, on the 5HT3 receptor and a judicious selection of them might contribute to reducing the incidence of PONV clinically.
Anxiety ; Baths ; Cholinergic Antagonists ; Clone Cells ; Cognition ; DNA, Complementary ; Electrodes ; Gallamine Triethiodide ; Incidence ; Nausea ; Neuromuscular Blockade* ; Neuromuscular Blocking Agents* ; Nociception ; Oocytes* ; Peripheral Nervous System ; Postoperative Nausea and Vomiting ; Receptors, Nicotinic ; RNA, Complementary ; Serotonin* ; Tubocurarine ; Vecuronium Bromide ; Vomiting ; Xenopus*

PURPOSE: Dioctahedral smectite is an alumina silicate of phyllitic structure and absorbs bile acid in the intestine, forming a non-absorbable complex preventing enterohepatic circulation. The purpose of this study is to clarify the value of dioctahedral smectite and the adequate dosage, in combination with phototherapy, as well as to confirm whether it shortens the duration of hospitalization and to compare dioctahedral smectite with cholestyramine. METHODS: Total 45 full-term neonate with a total bilirubin level greater than 12 mg/dl were studied. The neonate were randomly divided into three groups : 1) Only phototherapy group (A) 2) 3.0 g/day dioctahedral smectite with phototherapy group (B) 3) 1.0 g/kg/day cholestyramine with phototherapy group (C). RESULTS: The mean serum bilirubin level of group B and C decreased significantly compared to group A at 48, 72 and 96 hours after the beginning of the study. The duration of phototherapy and hospitalization significantly decreased in group B and C. CONCLUSION: The data revealed that oral administration of dioctahedral smectite not only increased the efficacy of phototherapy, but also shortened the duration of phototherapy and can substitute for cholestyramine.
Administration, Oral* ; Aluminum Oxide ; Bile ; Bilirubin ; Cholestyramine Resin* ; Enterohepatic Circulation ; Hospitalization ; Humans ; Hyperbilirubinemia, Neonatal* ; Infant, Newborn ; Intestines ; Jaundice, Neonatal ; Phototherapy* ; Silicates

Aspergillus funigatus and other pathogenic fungi synthesize a toxic epidithi-odiopiperzine (ETP) metabolite, namely gliotoxin. Gliotoxin commonly react with sulfhydryl groups, and then, forms hydrogen peroxide. These fungal toxins induce apoptotic cell death in various cells. Apoptosis induced by gliotoxin need calcium. Effect of calcium preconditioning was not reported in gliotoxin-induced apoptosis. To examine the effect of protein kinase C (PKC) and calcium which was regulate caspase-3, PKC and calcium preconditioning before gliotoxin treatment, apoptotic agents such as bcl-2 family, caspase-3 and DNA fragmentation in A7r5 cell line from rat smooth muscle cell were studied. These results showed that gliotoxin induces the expression of bad of bcl-2 family, caspase-3 activation and DNA fragmentation in A7r5 cells. Gliotoxin treatment followed by calcium and PKC preconditioning suppress the Bad of bcl-2 family, and inhibited caspase-3 activation, respectively. These results suggest that PKC and calcium preconditioning protect the gliotoxin-induced apoptosis, through the protection of pro-apoptotic bcl-2 family in A7r5 cells.
Animals ; Apoptosis ; Aspergillus ; Calcium ; Caspase 3 ; Cell Death ; Cell Line ; DNA Fragmentation ; Fungi ; Gliotoxin ; Humans ; Hydrogen Peroxide ; Muscle, Smooth* ; Mycotoxins ; Myocytes, Smooth Muscle* ; Protein Kinase C ; Rats*

Nitric oxide (NO) elevates intracellular calcium. But the actions of calcium in NO-induced cell death are not well understood. This study was carried out to investigate the signal transduction pathways of calcium and NO-induced cytotoxicity in H9c2 cardiac myoblasts by using NO donor compounds such as sodium nitroprusside (SNP) and S-nitroso-N-acetylpenicillamine (SNAP). Pretreatment of intracellular calcium chelating agent (BAPTA/AM) or L-type calcium channel blockers (nicardipine, nifedipine, diltiazem and veraparmil) or T-type calcium channel blocker (flunarizine) blocked SNP-induced cytotoxicity respectively only in a three hours. However, thapsigargin (TG), which inhibits endoplasmic reticulum dependent Ca(2+)-ATPase and thereby increases cytosolic Ca(2+), augmented SNP-induced cytotoxicity. The protective effect of BAPTA/AM was inhibited by treatment of protein synthesis inhibitor, cyclohexamide. In addition, pyrrolidine dithiocarbamate (PDTC), NF-kB inhibitor, attenuates the protective effect of BAPTA/AM against SNP-induced cytotoxicity. It is indicated that the protective effect of BAPTA/AM against NO-induced cytotoxicity might be due to the expression of protein related to activation of NFkB. From these results, it is concluded that SNP-induced cytotoxicity is mediated by calcium in a 3 hours via down regulation of protein expression rleated to activation of NFkB.
Calcium Channels, L-Type ; Calcium Channels, T-Type ; Calcium* ; Cell Death ; Cytosol ; Diltiazem ; Down-Regulation ; Endoplasmic Reticulum ; Humans ; Myoblasts, Cardiac* ; NF-kappa B ; Nifedipine ; Nitric Oxide ; Nitroprusside ; S-Nitroso-N-Acetylpenicillamine ; Signal Transduction* ; Thapsigargin ; Tissue Donors

Paclitaxel (taxol) is known as effective drug inhibition of cell cycle encouraging activity in human ovarian and metastatic breast cancers and malignant melanoma. It is an antimicrotubule agent that is believed to mediate its antineoplastic effects by inducing mitotic arrest followed by apoptosis. The relation between phorbol 12 myristate 13 acetate (PMA), protein kinase C (PKC) activator, and taxol-induced apoptosis is not well understood until now. This study was performed to investigate the effects of PMA on the signal transduction pathways of taxol-induced apoptosis in MCF-7 human breast carcinoma cells. Taxol-induced apoptosis is attenuated by curcumine, JNK inhibitor, and pyrrolidine dithiocarbamate (PDTC), inhibitor of NFkB. Pretreatment with PKC activator (PMA) or protein kinase A (PKA) activators (forskolin and dibutyryl cAMP) inhibited taxol-induced apoptosis in MCF-7 cells. In addition, thapsigargin, a specific inhibitor of endoplasmic reticulum(ER) Ca(2+)-ATPase and CaCl2, also blocked the activation of caspases by taxol. From these results suggest that taxol-induced apoptosis may be mediated via JNK or NFkB pathway and PKC activation.
Apoptosis ; Breast ; Breast Neoplasms ; Caspases ; Cell Cycle ; Curcumin ; Cyclic AMP-Dependent Protein Kinases ; Humans ; MCF-7 Cells* ; Melanoma ; Myristic Acid ; Paclitaxel ; Protein Kinase C ; Signal Transduction ; Thapsigargin

Aspergillus funigatus and other pathogenic fungi synthesize a toxic epidithi- odiopiperzine (ETP) metabolite called gliotoxin. Gliotoxin is an epidithiodiopiperzine compound which can both react with sulfhydryl groups and form hydrogen peroxide. The fungal toxin gliotoxin induces apoptotic cell death in a variety of cells. Apoptosis induced by gliotoxin need calcium but effect of calcium preconditioning is unknown by gliotoxin. We studied the effect of protein kinase C and calcium preconditioning on gliotoxin-induced apoptosis in H9c2 cell. PKC and calcium preconditiong inhibited DNA fragmentation by gliotoxin. From this above results suggest that gliotoxin induce apoptosis via caspase-3 activation, because caspase-3 inhibitor (DEVD-CHO) didn't induce apoptosis in gliotoxin treated H9c2 clls. Calcium and PKC preconditioning inhibit caspase-3 activation by gliotoxin. These data means that PKC preconditioning is related with caspase-3 regulate in gliotoxin-induced apoptosis.
Apoptosis* ; Aspergillus ; Calcium ; Caspase 3 ; Cell Death ; DNA Fragmentation ; Fungi ; Gliotoxin* ; Hydrogen Peroxide ; Protein Kinase C* ; Protein Kinases*

Nitric oxide (NO) is mainly involved in brain ischemic damage to elucidate the protective mechanism of NO pretreatment on ischemic-induced cytotoxicity. This study was investigated whether NO pretreatment inhibits the increase of iNOS expression by lipopolysaccharide (LPS) combined phorbol 12-myristate 13-acetate (PMA) via regulating NF-kB activation in C6 glial cells. C6 glial cells with LPS and PMA for 72 hours markedly induced NO, but sodium nitroprusside (SNP) (100 nM) pretreatment before exposure of LPS and PMA significantly supressed NO production, iNOS expression and NF-kB activation by LPS and PMA. In addition, LPS and PMA treatment for 72 hours induced severely cell death and LDH release from cell into media in C6 glial cells. However SNP pretreatment before treatment of LPS and PMA significantly protected LPS and PMA induced cytotoxicity. Treatment with LPS and PMA induced caspase 3 activation follewed by chromosomal condensation, and fragmentation of nuclei in C6 glial cells. SNP pretreatment before exposure to LPS and PMA supressed caspase 3 activation and inhibited chromosomal condensation and fragmentation of nuclei. From these above results, it is suggest that the protective effects of SNP pretreatment against LPS and PMA induced cytotoxicity may be mediated by inhibiting the expression of iNOS via regulating NF-kB activation.
Brain ; Caspase 3 ; Cell Death ; Neuroglia ; NF-kappa B ; Nitric Oxide* ; Nitroprusside