No abstract available.
Drug Interactions*

Using murine immortalized microglial cells (BV-2), we examined the regulation of RANTES production stimulated by lipopolysaccharide (LPS), focusing on the role of mitogen-activated protein kinase (MAPK) and nuclear factor (NF)-kappaB. The result showed that RANTES (regulated upon activation of normal T cell expressed and secreted) was induced at the mRNA and protein levels in a dose- and time-dependent manner in response to LPS. From investigations of second messenger pathways involved in regulating the secretion of RANTES, we found that LPS induced phosphorylation of extracellular signal-regulated kinase (Erk), p38 MAPK and c-Jun-N-terminal kinase (JNK), and activated NF-kappaB. To determine whether this MAPK phosphorylation is involved in LPS-stimulated RANTES production, we used specific inhibitors for p38 MAPK and Erk, SB 203580 and PD 98059, respectively. LPS-induced RANTES production was reduced approximately 80% at 25 micrometer of SB 203580 treatment. But PD 98059 did not affect RANTES production. Pyrrolidine-dithiocarbamate (PDTC), NF-kappaB inhibitor, reduced RANTES secretion. These results suggest that LPS-induced RANTES production in microglial cells (BV-2) is mainly mediated by the coordination of p38 MAPK and NF-kappaB cascade.
Chemokine CCL5* ; Microglia* ; NF-kappa B* ; p38 Mitogen-Activated Protein Kinases* ; Phosphorylation ; Phosphotransferases ; Protein Kinases ; RNA, Messenger ; Second Messenger Systems

BACKGROUND: Although nucleotides -like Adenosine Triphosphate (ATP) and its derivatives Adenosine, were known to induce growth inhibition and apoptosis in diverse cell lines, little is known about their effects on trophoblast. OBJECTIVE: To elucidate the effects of extracellular ATP and adenosine on trophoblast cell growth and to delineate if apoptosis is involved in this mechanism. MATERIALS AND METHODS: We used TL cell line, derived from human term placenta. The cells were cultured for 24, 48, and 72 hours after being treated with ATP and adenosine, each. Also, cell growth according to different concentrations of ATP and adenosine was evaluated. To test whether apoptosis was induced by each nucleotide, DNA fragmentation and nuclear condensation by Hoechst 33258 stain and P53 protein expression were evaluated. RESULTS: Cell growth was inhibited by ATP and adenosine in time and dose-dependent manner. Furthermore, the growth inhibitory effect of adenosine was stronger than ATP, whereas signs of DNA fragmentation and nuclear condensation were observed in ATP treated cells, but not in adenosine treated ones. CONCLUSION: Our results shows that ATP and adenosine exert inhibitory effect on growth in TL cell line. These findings suggest that pathological production of ATP or its metabolites, adenosine, may lead to a pathologic status such as preeclampsia or intrauterine growth restriction.
Adenosine Triphosphate* ; Adenosine* ; Apoptosis* ; Bisbenzimidazole ; Cell Line* ; DNA Fragmentation ; Humans* ; Nucleotides ; Placenta ; Pre-Eclampsia ; Trophoblasts

OBJECTIVES: To evaluate an association between depression and altered immunity, we examined peripheral T lymphocyte or natural killer (NK) cell measures plasma ACTH and cortisol using the flow cytometry in acute and unmedicated patients with major depressive disorder (MDD). METHODS: Forty-two patients with MDD from the outpatient clinic and forty normal controls from the hospital staff were recruited. We applied Hamilton Rating Scale for Depression (HAM-D) and Hamilton Rating Scale for Anxiety (HAM-A) for depressed subjects. Peripheral T lymphocyte or NK cell measures (CD3, CD4, CD8, or CD56) and plasma hormones (ACTH and cortisol) were obtained from all subjects. RESULTS: There were no statistical differences in CD3, CD4, CD8, or CD56 between the two subjects. The number of CD56 cells negatively correlated with HAM-D scores (r=-0.42, p<0.01), but did not correlate with HAM-A scores in patients with MDD. The number of CD56 cells showed strong negative correlation with CD4/CD8 (r=-0.47, p<0.01) in the control group, but not in the depressed group. Patients with MDD had higher cortisol level than controls within the normal range. CONCLUSION: The trait of immunological imbalance and HPA axis abnormality were shown in patients with MDD. Especially, the severity of depression, but not the anxiety, could be reflected as decreased number of CD56 (NK T) cells in acute and unmedicated state.
Adrenocorticotropic Hormone ; Ambulatory Care Facilities ; Anxiety ; Axis, Cervical Vertebra ; Depression ; Depressive Disorder, Major* ; Flow Cytometry ; Humans ; Hydrocortisone ; Killer Cells, Natural ; Lymphocytes ; Natural Killer T-Cells* ; Plasma ; Reference Values

A hippocalcin cDNA from rat brain cDNA library was amplified by polymerase chain reation(PCR) and cloned using TA Cloning technique. For this PCR cloning, 29mer and 28mer oligonucleotide primers containing BamHl and EcoRl sites at the 5' end and 3' end, respectively were used. The nucleotide sequence of hippocalcin cDNA c1one was determined, and the complete amino acid sequence was deduced. Recombinant clone contained a cDNA insert of 610 base pairs with 582 nucleotides of open reading frame including the temination codon, 23 nucleotide of 5'-untranslated region, and 5nucleotides of 3'-nutran,slated region. The open reading frame encoded a polypepetid comprising 193 amino acids with molecular weight of 22kDa. The cDNA insert was subcloned into pVLI393 Baculovirus transfer vector. The recombinant hippocalcin was expressed in insect cell(Sf9 cell) using expression vector pVL1393. The hippocalcin expressed was purified as a single band on polyacrylamide gel electrophoresis following hydrophobic phenyl HPLC and TSKgel G3000SW gel filtration HPLC. Molecular size of rat brain hippocalcin protein expressed in this system was estimated to be 22kDa. Myristoylated hippocalcin migrated faster than nonmyristoyated form on SDS-polyacrylamide gel. Less than 10% of total hippocalcin expressed was myristoylated in this baculovirus expression system.
Amino Acid Sequence ; Amino Acids ; Animals ; Baculoviridae ; Base Pairing ; Base Sequence ; Brain* ; Chromatography, Gel ; Chromatography, High Pressure Liquid ; Clone Cells ; Cloning, Organism ; Codon ; DNA Primers ; DNA, Complementary* ; Electrophoresis, Polyacrylamide Gel ; Gene Library ; Hippocalcin* ; Insects ; Molecular Weight ; Nucleotides ; Open Reading Frames ; Polymerase Chain Reaction ; Rats*

BACKGROUND: Atopic dermatitis (AD) is a common inflammatory skin disease. Atopic dermatitis is associated with increased IL-4, IL-5, IL-10 and IL-13 but decreased INF-gamma and TNF production. IL-10 production has been implicated in autoimmunity because of its effect on B-cell proliferation and antibody production. The study of IL-10 gene polymorphism is of interest because of the pivotal role of IL-10 in the regulation of inflammatory and immune responses. TNF-beta significantly upregulates INF-gamma but downregulates IL-5, IL-13 and IgE, which suggests a potential role of TNF-beta in the pathogenesis of atopic dermatitis. OBJECTIVES: We have investigated polymorphism of IL-10 promoter gene and TNF-beta gene. METHODS: Seventy one patients with atopic dermatitis and one hundred and sixty six normal subjects participated in this study with analysis of polymorphism of IL-10 promoter (-1082), (-819) gene and seventy one patients with atopic dermatitis and one hundred and forty one normal subjects participated in the analysis of polymorphism of TNF-beta gene. The patients in this study were recently diagnosed with atopic dermatitis. RESULTS: The frequency of IL-10 promoter (-1082) genotypes (A/A, A/G, G/G), genes (A, G), IL-10 promoter (-819) genotypes (T/T, T/C, C/C) and genes (T, C) did not show any significant difference between atopic dermatitis patients and normal controls. There was no significant difference in the frequency of TNFB genotypes (TNFB*1/TNFB*1, TNFB*1/TNFB*2, TNFB*2/TNFB*2) and genes (TNFB*1, TNFB*2) in patients of atopic dermatitis and normal controls. CONCLUSION: The results of this study suggest that the other regions of the IL-10 promoter gene and TNFB gene should be investigated for polymorphism of atopic dermatitis. And the difference of IL-10 promoter and TNFB gene polymorphism between caucasian and Korean needs to be evaluated.
Antibody Formation ; Autoimmunity ; B-Lymphocytes ; Dermatitis, Atopic* ; Genotype ; Humans ; Immunoglobulin E ; Interleukin-10* ; Interleukin-13 ; Interleukin-4 ; Interleukin-5 ; Lymphotoxin-alpha* ; Polymorphism, Genetic* ; Skin Diseases

OBJECTIVES: It is known that the high fibrogenecity of particles is connected with their cytotoxicity for macrophages. Although the molecular mechanism leading to fiber-induced fiber-induced cytotoxicity is still not clear, several mechanism have been suggested. The release of reactive oxygen species (ROS) from activated alveolar macrophages (AM) by dust have been suggested as a possible mechanism of particle-induced cell damage. But the mechanism which man-made vitreous fiber (MMVF) induces the production of ROS in AM is still not clear. In this study, we evaluated the relationship between ROS production and lactate dehydrogenase (LDH) release from alveolar treated with refractory ceramic fiber (RF2) or rock wool (RW1) and signal transduction path-way of ROS production in RF2 or RW1 exposed AM. METHODS: We investigated LDH release from MMVF-stimulated AM for index of cytotoxicity. To determine what kind of signal transduction pathways are involved in MMVF-stimulated ROS generation, we used some drugs which have an effect on the signal transduction pathway. RESULTS: RF2 and RW1 induced increase of LDH release with dose-dependent manner with RF2 having greater effect than RW1. There was a dose-dependent increase in the production of ROS by RF2 or RW1. At all level of concentration,. RF2 induced more ROS production than RW1. Inhibitors of PKC (bisindolylmaleimide), PLC (U73122 and neomycine) and PTK (genistein and erbstatin) suppressed RF2 or RW1-induced ROS production. CONCLUSION: There was significant correlation between LDH release and ROS production from AM treated with RF2 or RW1. RF2 and RW1 induced ROS generation through protein kinase C (PKC), phospholipase C (PLC) and protein tyrosin kinase (PTK) pathways.
Ceramics* ; Dust ; L-Lactate Dehydrogenase ; Macrophages ; Macrophages, Alveolar* ; Phosphotransferases ; Protein Kinase C ; Reactive Oxygen Species* ; Signal Transduction* ; Type C Phospholipases ; Wool

Abstract Phospholipase D (PLD) plays an important role as an effector in a variety of physiological processes that reveal it to be a member of the signal transducing phospholipases. Recently, PLD2 was reported as a necessary intermediate in preventing apoptosis induced by hydrogen peroxide or hypoxia in rat pheochromocytoma (PC12) cells. The data presented here show that both PLD isozymes, PLD1 and PLD2 are also required in attenuating glutamate-induced cell death in PC12 cells. Treatment of PC12 cells with glutamate resulted in induction of apoptosis in these cells, which is accompanied by decreased PLD activity and increased ceramide concentration. Incubation of PC12 cells with exogenous C6-ceramide showed a time-dependent decrease of PLD activity. When cDNAs of PLD1 and PLD2 were transfected into PC12 cells respectively, overexpression of PLD1 or PLD2 resulted in inhibition of glutamate-induced apoptotic cell death. These data indicate that both PLD1 and PLD2 play a protective role against glutamate-induced cell death in PC12 cells.
Animals ; Apoptosis/drug effects/*physiology ; Cell Survival/drug effects ; Ceramides/pharmacology ; Dose-Response Relationship, Drug ; Enzyme Activation ; Gene Expression Regulation, Enzymologic/drug effects ; Glutamic Acid/*toxicity ; Isoenzymes/drug effects/genetics/*metabolism ; Kinetics ; PC12 Cells ; Phospholipase D/chemistry/drug effects/genetics/*metabolism ; Rats ; Sphingolipids/metabolism

In the freshly isolated rat nephron, the effect of endothelin-1, -2 and -3 (ET-1, -2 and -3) on cytosolic free calcium concentration ((Ca2+)i) was determined using the fluorescent indicator Fura-2/AM. (Ca2+)i increase was investigated in 9 parts of the single nephron including glomerulus (Glm), S1, S2, S3, Cortical and medullary thick ascending limb and cortical (CCT) and outer medullary collecting tubule (OMCT). Endothelins increased (Ca2+)i in Glm (ET-1; 127+/-17%, ET-2; 93+/-5%, ET-3; 169+/-17%), CCT (ET-1; 30+/-6%, ET-2; 38+/- 19%, ET-3; 158+/-18%) and OMCT (ET-1; 197+/- 11%, ET-2; 195+/- 11%, ET-3; 215+ 37%) at 10(-7) M. In OMCT, ET-1 and ET-2 increased (Ca2+)i in a dose-dependent manner (10(-10) ~ 10(-6) M). To the contrary, ET-3-induced (Ca2+)i rise was begun from 10(-12) M. BQ-123Na, an antagonist of ETA receptor, at 10(-4) M inhibited about 30% of (Ca2+)i rise induced by ET-1 and -3. Binding experiments using (125I)ET-3 showed the existence of ETB receptor in OMCT. This binding was replaced by ET-1, ET-2 or ET-3 by the almost same degree but not by angiotensin II or vasopressin.
Angiotensin II ; Animals ; Calcium* ; Cytosol ; Endothelin-1 ; Endothelin-2 ; Endothelins* ; Extremities ; Nephrons* ; Rats* ; Vasopressins

PURPOSE: Febrile seizures are characterized by a heterogenous phenotype segregating as an autosomal dominant trait with incomplete penetrance. Mutations in GABRG2 gene were identified in two families with generalized epilepsy and febrile seizures plus (GEFS+) and with absence epilepsy and febrile seizures(FSs). The present study assessed the role of GABRG2 gene in FSs and GEFS+ of the Korean population. METHODS: 66 FSs, 20 GEFS+ and 94 healthy control subjects were selected throughout a collaborative study of Catholic Child Neurology Research Group. The SNP211037 of GABRG2 was screened by DHPLC. DNA fragments showing variant chromatograms were subsequently sequenced. Genotypes and allelic frequencies for GABRG2 gene polymorphism in three groups were compared. RESULTS: The number of individuals with the GABRG2(SNP211037)-C/C genotype in patients with FSs was significantly greater compared with that in healthy control subjects and the GABRG2(SNP211037)-C allele frequency in patients with FSs was significantly higher than that in healthy control subjects. The odds ratio for developing FSs in individuals with the GABRG2(SNP211037)-CC genotype was 5.96 compard with individuals with the GABRG2(SNP211037)-T/T genotype. In contrast, the GABRG2 (SNP211037) gene in GEFS+ and control groups was not significantly different. CONCLUSION: Theses data suggest that genomic variations of GABRG2 might be one of the susceptibility factors for FSs in the Korean population.
Child ; DNA ; Epilepsy, Absence ; Epilepsy, Generalized ; Gene Frequency ; Genotype ; Humans ; Neurology ; Odds Ratio ; Penetrance ; Phenotype ; Polymorphism, Single Nucleotide* ; Seizures, Febrile*