Cytosine arabinoside(AraC) inhibits DNA synthesis and beta-DNA polymerase, an enzyme involved in DNA repair. This a potent antimitotic agent, is clinically used as an anticancer drug with side effect of severe neurotoxicity. Earlier reports suggested that inhibition of neuronal survival by AraC in sympathetic neuron may be due to the inhibition of a 2'-deoxycytidine-dependent process that is independent of DNA synthesis or repair and AraC induced a signal that is triggers a cascade of new mRNA and protein synthesis, leading to apoptotic cell death in cultured cerebellar granule cells. The present study would suggest whether caspase family(ICE/CED-3-like protease) involved in AraC-induced apoptosis pathway of PC12 cells. It was observed that treatment of PC12 cells with AraC led to decrease of viability by MTT assay and morphology changes, which did not suggest that AraC induced apoptosis in PC12 cells. The mRNA of caspase-1/caspase-3 were expressed in PC12 cells constitutively, and AraC did not activate caspase family. These results suggest that caspase-1/caspase-3 may not be required for AraC-induced cell death pathway in PC12 cells.
Animals ; Apoptosis ; Cell Death ; Cytosine* ; DNA ; DNA Repair ; Humans ; Neurons ; PC12 Cells* ; RNA, Messenger

The cAMP-dependent protein kinase(PKA) is an intracellular enzyme with serine-threonine kinase activity that plays a key role in cell growth, differentiation, and apoptosis in eukaryotes. In order to understand the PKA signal transduction pathway regulating cell life cycle and identify its role, we focused on the characterization of up-/down-regulated genes by PKA using the differential display polymerase chain reaction. Seven differentially expressed sequence tags(DEST) have been obtained. Among these DESTs, 2DESTs were homologous to the sequence of genes from BLAST search result. KC1-5 DEST that was up-regulated in A123.7 cells was highly corresponded to mouse apoptosis-related gene(MA-3) or mouse mRNA for topoisomerase inhibitor suppressed(TIS). MA-3 was induced in various types of apoptosis, specially in NGF-deprived apoptotic PC12 cells, TIS was down-regulated in the RVC lymphoma cells incubated with topoisomerase inhibitor that induces DNA strand breakages. PG1-1DEST that was highly expressed in PC12 cells was corresponded to transposon Tn103'-end. Tnansposon Tn10 was up-regulated in differentiated myeloblastic ML-1 cells by 12-O-tetradecanoylphorbol-13-acetate. This study illuminates that MA-3/TIS was down-regulated by PKA activity, and transposon Tn10 was up-regulated by it.
Animals ; Apoptosis ; DNA ; Eukaryota ; Granulocyte Precursor Cells ; Life Cycle Stages ; Lymphoma ; Mass Screening* ; Mice ; PC12 Cells* ; Polymerase Chain Reaction ; Protein-Serine-Threonine Kinases ; RNA, Messenger ; Signal Transduction

The Bcl-2 protein has been shown to block apoptosis induced by a variety of stimuli. We have performed the experiments which cell death can be blocked by the bcl-2 proto-oncogene under moderate(50-100mM) or high ethanol treatment(400-600mM). As a result of morphological changes, and MTT assay, cell death was blocked by Bcl-2 under 100mM ethanol. However, the results of DNA fragmentation and RT-PCR(ICE, and CPP32), immunoblotting(CPP32, and PARP) for SK-pcDNA3 cells(vector only) and SK-Bcl-2 cells(stably expressed bcl-2 gene) were showen to be no significant differences between two cell lines. These result suggested that cell death induced by ethanol was not followed by apoptosis mechanism, and was blocked by the bcl-2 proto-oncogene with moderate ethanol.
Apoptosis ; Cell Death* ; Cell Line ; DNA Fragmentation ; Ethanol* ; Ice ; Proto-Oncogenes*

An association study with Korean alcoholic patients(n=50) and normal controls(n=53) was performed to find the relationship between catechol-O-methyltransferase(COMT) gene polymorphism and alcoholism using polymerase chain reaction-restriction fragment length polymorphism. When we compared the allele and genotype frequencies of Nla III COMT gene polymorphism in alcoholism and normal controls, there was no significant difference between two groups. Our results do not support an association between the Nla III polymorphism of COMT gene and alcoholism.
Alcoholics ; Alcoholism* ; Alleles ; Genotype ; Humans

This study was designed to examine the effects of two antidepressant drugs on the expression of c-fos mRNA in cultured embryonic rat hippocampal neurons. The drugs used were imipramine and amitriptyline. On the fourth day of culture, hippocampal neurons were treated with variable concentrations of each drug. Competitive RT-PCR(Reverse Transcriptase-PCR) analysis was used to quantify the c-fos mRNA expression induced by each drug. Experimental results showed that acute and direct treatment with imipramine and amitriptyline with relatively low concentrations(imipramine < or =10micrometer, amitriptyline < or =10micrometer) had no inductive effect on the expression of c-fos mRNA in the rat hippocampal neurons. However, after treatment with relatively high concentrations(imipramine > or =100micrometer, amitriptyline > or =100micrometer) c-fos mRNA was not detected. These findings suggest the followings. Firstly, the action mechanisms of these drugs on the hippocampal neurons might not be mediated by c-fos but by other immediate-early genes(IEGs). Secondly, their actions may be mediated indirectly via other areas of the brain. Thirdly, the expression of c-fos might be inhibited by high concentrations of these drugs, or the high concentrations could induce cell death. Finally, though cell death remains to be confirmed, the inhibition of c-fos induction or cell death could play a role in the cognitive impairments known to be adverse effects of some antidepressants. This study is believed to be a first step toward understanding the mechanisms of learning and memory. Further studies are needed to investigate the expression of various IEGs and changes in the hippocampal neurons of rat resulting from chronic treatment with antidepressant drugs.
Amitriptyline ; Animals ; Antidepressive Agents* ; Brain ; Cell Death ; Imipramine ; Learning ; Memory ; Neurons* ; Rats* ; RNA, Messenger*

Lymphedema is not uncommon, but it can often be undiagnosed until discomfort or complications occur. It tends to develop slowly, but is progressive without proper treatment. Lymphedema occurs when the lymphatic fluid load is greater than the ability of transport, resulting not only in excessive accumulation of tissue fluid but also in deformity of appearance, immobility, and more serious consequences. Stage I lymphedema can be improved by simply promoting drainage with elevation and compression garments. Stage II or III lymphedema should be managed intensively with complete decongestive therapy using a combination of skin care, exercise, elevation, manual lymph drainage, intermittent pneumatic compression, multilayer lymphedema bandaging and weight reduction. The safety and effectiveness of other treatment modalities for lymphedema such as liposuction, microsurgical lymphatic reconstruction, needle aspiration, stem cells, laser therapy, and iliac vein stenting should further be investigated. Since lymphedema is progressive, the diagnosis and treatment of lymphedema at the earliest possible stage is very important. Complete decongestive therapy is principal, and psychosocial support is an important element of the treatment of lymphedema.
Congenital Abnormalities ; Diagnosis* ; Drainage ; Iliac Vein ; Laser Therapy ; Lipectomy ; Lymphedema* ; Needles ; Skin Care ; Stem Cells ; Stents ; Weight Loss

Alcohol influences the neuroadaptation of brain cells where receptors and enzymes like protein kinase C (PKC) exist. Naltrexone acts on opioid receptors. However, other mechanisms of action remain unknown. We prepared SH-SY5Y neuroblastoma cells, and fed them with 150 mM ethanol for 72 h followed by treatment with naltrexone for 24 h. We performed microarray analysis and reverse transcriptase-polymerase chain reaction. Our results showed that PKC epsilon increased 1.90 times and showed an overall decreasing pattern as time increased. Phosphorylated ERK also increased 2.0 times according to the change of PKC epsilon. Integrin alpha7 increased 2.32 times and showed an increasing pattern as time increased. In conclusion, naltrexone influences PKC epsilon neuronal signaling system and endothelial adhesion molecule integrin alpha7 in addition to the well-known opioid system.
Antigens, CD/*metabolism ; Cell Line, Tumor ; Comparative Study ; DNA, Complementary/genetics ; Humans ; Integrin alpha Chains/*metabolism ; Naltrexone/*pharmacology ; *Neuroblastoma/enzymology/metabolism/pathology ; Oligonucleotide Array Sequence Analysis ; Protein Kinase C-epsilon/*metabolism ; Research Support, Non-U.S. Gov't ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors

OBJECTIVE: The aim of this study was to examine the effect of dexamethasone and fluoxetine on the expression of 70 kDa heat shock protein (HSP70) in C6 glioma cells. METHODS: The C6 glioma cells belong to control group were incubated with DMEM culture solution, the cells belong to dexamethasone group were incubated with dexamethasone for 6 hours, and the cells belong to fluoxetine group were incubated with fluoxetine for 1, 6, 24, and 72 hours, separately, and then exposed to dexamethasone for an additional 6 hours. Crude extracts from control, dexamethasone and fluoxetine-treated C6 glioma cells were separated on a 10% SDS-PAGE and probed with anti-HSP70 mAb. RESULTS: 1) Dexamethasone (10 uM, 6 hours) reduced the level of HSP70 expression relative to control, but this reduction was not statistically significant. 2) Pretreatment with fluoxetine (10 uM, 1, 6, 24, and 72 hours) and exposure to dexamethasone (10 uM, 6 hours) decreased the level of HSP70 expression according to the duration of fluoxetine treatment. 3) Fluoxetine significantly reduced the level of HSP70 at 24 and 72 hours compared to control. However, compare to the level of HSP70 expression at 24 hours, the level of HSP70 expression at 72 hours was elevated. CONCLUSION: These findings suggest that dexamethasone and fluoxetine may affect HSP70 expression through effects on GR.
Animals ; Complex Mixtures ; Dexamethasone ; Electrophoresis, Polyacrylamide Gel ; Fluoxetine* ; Glioma* ; Heat-Shock Proteins* ; Hot Temperature* ; HSP70 Heat-Shock Proteins* ; Rats*

OBJECTIVE: The aim of this study was to examine the effect of dexamethasone and fluoxetine on the expression of 70 kDa heat shock protein (HSP70) in C6 glioma cells. METHODS: The C6 glioma cells belong to control group were incubated with DMEM culture solution, the cells belong to dexamethasone group were incubated with dexamethasone for 6 hours, and the cells belong to fluoxetine group were incubated with fluoxetine for 1, 6, 24, and 72 hours, separately, and then exposed to dexamethasone for an additional 6 hours. Crude extracts from control, dexamethasone and fluoxetine-treated C6 glioma cells were separated on a 10% SDS-PAGE and probed with anti-HSP70 mAb. RESULTS: 1) Dexamethasone (10 uM, 6 hours) reduced the level of HSP70 expression relative to control, but this reduction was not statistically significant. 2) Pretreatment with fluoxetine (10 uM, 1, 6, 24, and 72 hours) and exposure to dexamethasone (10 uM, 6 hours) decreased the level of HSP70 expression according to the duration of fluoxetine treatment. 3) Fluoxetine significantly reduced the level of HSP70 at 24 and 72 hours compared to control. However, compare to the level of HSP70 expression at 24 hours, the level of HSP70 expression at 72 hours was elevated. CONCLUSION: These findings suggest that dexamethasone and fluoxetine may affect HSP70 expression through effects on GR.
Animals ; Complex Mixtures ; Dexamethasone ; Electrophoresis, Polyacrylamide Gel ; Fluoxetine* ; Glioma* ; Heat-Shock Proteins* ; Hot Temperature* ; HSP70 Heat-Shock Proteins* ; Rats*

Nephrogenic adenoma is rare, especially in the ureter. It is thought to represent a metaplastic phenomenon forming the tubules which resemble renal tubules. The cause is unknown but may be associated with chronic irritations such as surgical trauma, incarcerated calculi and infection. We report a case of nephrogenic adenoma of the ureter in a 44-year-old woman who bad been suffered from the left flank pain due to left ureteral stone with giant hydronephrosis.
Adenoma* ; Adult ; Calculi ; Female ; Flank Pain ; Humans ; Hydronephrosis ; Ureter*