Nitric oxide (NO) induces apoptotic cell death in murine RAW 264.7 macrophages. To elucidate the roles of SEK1/MKK4, a upstream kinase for both c-Jun N-terminal kinase/stress-activated protein kinase (JNK/SAPK) and p38 kinase, on NO-induced apoptosis, we generated clones of RAW 264.7 cells which stably overexpressd kinase inactive SEK1 (RAW/SEK1-Kl) or wild type SEK1 (RAW/SEK1-WT). Treatment of kinase inactive SEK1 transfected RAW 264.7 cells (RAW/SEK1-Kl) with sodium nitroprusside (SNP), a NO generating agent, significantly decreased the cell viability up to 20% of RAW control cells which were treated with the same amount of SNP. However, RAW/SEK1-WT cells were less susceptible to NO induced apoptosis. For a while, caspase-3 like activity in NO treated RAW/SEK1-Kl cells was significantly increased with parallell to apoptotic death rate. However, caspase1 like activity was not affected by NO in any transfectants. The NO induced apoptosis in RAW/SEK1-Kl cells was significantly prevented by the addition of caspase-3 like inhibitor (N-Ac- DEVD-CHO). In addition, the phosphotransferase activity of JNK1 in NO-treated RAW/SEK1-WT is significantly increased, but not in RAW/SEK1-Kl cells. These results suggest that SEK1 may play anti-apoptotic role in RAW cells from NO-induced apoptosis.
Apoptosis ; Caspase 3 ; Cell Death ; Cell Survival ; Clone Cells ; Macrophages ; Mortality ; Nitric Oxide* ; Nitroprusside ; Phosphotransferases ; Protein Kinases

Actinic kratosis can develop dnring PUVA therapy especially at high accumulative doses(more than 1000Joules/cm) but it can also develop at low accumulative doses(lese than 200Joules/cm). PUVA therapy should be done carefully and the dermatologist has to observe the patients skin more closely to detect development of the keratosis. We present a case of multiple actinic keratoses that developed during PUVA therapy in a 53 year old vitiligo patient. They developed during relatively low accumulative dose PUVA therapy (827Joules/cm2).
Actins* ; Humans ; Keratosis ; Keratosis, Actinic* ; Middle Aged ; PUVA Therapy* ; Skin ; Vitiligo*

PURPOSE: Recent studies indicate that widely used chemotherapeutic agents induce apoptosis in susceptible cells. One of the effector arms in this cell death pathway is composed of cysteine proteases belonging to the caspase family. In cells, caspase-3 has been shown to play an important role as a downstream member of protease cascade, where various cell death pathways converge into the same effector pathway. JNK, a member of the mitogen-activated protein kinase pathway, is activated in response to many stressful stimuli including heat shock, UV irradiation, protein synthesis inhibitors, and inflammatory cytokines. In this study, we investigated whether JNK1 & caspase-3 play a role in the apoptosis induced by adriamycin (ADR). METHODS: U937 cells were cultured in RPMI 1640 and treated with different concentrations of ADR. Cellular DNA was extracted and analyzed by electrophoresis on a 1.5% agarose gel to detect DNA fragmentation. The activity of caspase-3 was measured by the proteolytic cleavage of the fluorogenic substrate DEVD-AMC. The activity of JNK1 was measured by in vitro immunocomplex kinase assay with 2 microgram of GST-c Jun as a substrate and quanititated using phosphoimager analyzer. RESULTS: ADR induced the apoptotic death of U937 myeloid cells in a dose-dependent manner, which was characterized by increasing ladder-pattern DNA fragmentation. Consistent with apoptotic death of U937 cells, ADR induced the catalytic activation of caspase-3 as well as JNK1 at 2.5 microgram/mL of concentrations. CONCLUSION: Adriamycin induces apoptosis of human myeloid leukemic U937 cells via activation of caspase-3 and cJun-N terminal kinase1 (JNK1)/Stress activated protein kinase (SAPK).
Apoptosis* ; Arm ; Caspase 3* ; Cell Death ; Cysteine Proteases ; Cytokines ; DNA ; DNA Fragmentation ; Doxorubicin* ; Electrophoresis ; Fluorescent Dyes ; Hot Temperature ; Humans* ; Myeloid Cells ; Phosphotransferases ; Protein Kinases ; Protein Synthesis Inhibitors ; Sepharose ; Shock ; U937 Cells*

BACKGROUND: Chronic myelogenous leukemia is a chronic myeloproliferative disorder characterized by leukocytosis with myeloid elements at all stages of differentiation, t(9;22)(q34;q11) and bcr/abl rearrangement. We studied hydroxyurea induced apoptotic changes such as externalization of phosphatidylserine, caspase activities on human chronic myelogenous leukemic cell line, K562 cells. METHODS: K562 cells were grown in RPMI 1640 supplemented with 10% fetal bovine serum and treated hydroxyurea. Viability was examined by MTT assay. Apoptosis were examined by annexin V stain, caspase (such as caspase-, caspase-, caspase-, caspase-, and caspase-) activities, and DNA fragmentation. RESULTS: The viability of K562 cells were markedly decreased in a dose dependent manner of hydroxyurea. Phosphatidylserine externalization was detected by annexin V stain after 3 hours in hydroxyurea treated K562 cells and the value of lactate dehydrogenase was not significantly changed in their culture media. The upstream effector of caspase- was slightly increased and had influenced on caspase-. And downstream acting caspase protease of caspase- was markedly increased in a time dependent manner at hydroxyurea treated K562 cells. In addition, however the activities of caspase- and caspase- were not increased. We also found DNA fragmentation at hydroxyurea treated K562 cells between 48 hours and 72 hours on agarose gel electrophoresis. CONCLUSIONS: Hydroxyurea induces apoptotic change in K562 cells via externalization of phosphatidylserine, activations of caspase-, caspase-, caspase- proteases, and DNA fragmentation.
Annexin A5 ; Apoptosis ; Cell Line* ; Culture Media ; DNA Fragmentation ; Electrophoresis, Agar Gel ; Humans ; Hydroxyurea* ; K562 Cells* ; L-Lactate Dehydrogenase ; Leukemia, Myelogenous, Chronic, BCR-ABL Positive ; Leukocytosis ; Myeloproliferative Disorders ; Peptide Hydrolases

BACKGROUND/AIMS: The hepatitis B virus (HBV) genome contains binding sites for hepatocyte nuclear factors (HNF) 3 and 4 in the core domain of enhancer 1 (Enh1), and mutations in this domain have a strong impact on virus replication. We aimed to identify frequent base-mutation sites in the core domain of Enh1 and to examine the impact of these mutations on viral replication. METHODS: We studied virological characteristics and genetic sequences in 387 patients with chronic hepatitis B. We evaluated functional differences associated with specific mutations within the core domain of Enh1. RESULTS: Mutations in the core domain were found with significant frequency in C1126 (122/387 [31.5%], the binding site for HNF3) and in C1134 (106/387 [27.4%], the binding site for HNF4). A single mutation at nt 1126 (C1126) was identified in 17/123 (13.8%), and 105/123 (85.4%) had double mutations (C1126/1134). The level of HBV DNA (log10 copies/mL) was lower in single mutants (C1126, 5.81+/-1.25) than in wild (6.80+/-1.65) and double mutants (C1126/1134, 6.81+/-1.54). Similarly, the relative luciferase activity of C1126 and C1126/C1134 was 0.18 and 1.12 times that of the wild-type virus, respectively. CONCLUSIONS: Mutations in the HNF3 binding site inhibit viral replication, whereas mutations at the HNF4 binding site restore viral replication.
Binding Sites ; DNA ; Genome ; Hepatitis B virus ; Hepatitis B, Chronic ; Hepatitis, Chronic ; Hepatocyte Nuclear Factors ; Humans ; Luciferases ; Virus Replication ; Viruses

Adriamycin (ADR) is a potent anticancer drug that causes often severe cardiomyopathy. Previous reports have demonstrated that zinc accumulation is shown in rat myocardial cells following ADR treatment. However, the mechanism and role of zinc accumulation in ADR-induced cardiomyopathy are not yet elucidated. Zinc may be one of the key executors in ADR-induced cardiomyopathy. To test this hypothesis, we examined the cytotoxic effects of zinc on various cell lines including H9c2 cardiomyoblast cells, HL-60, U937, and C(6)-glial cells. Zinc induced significant the death of H9c2 cells at 0.125 mM in a dose-dependent manner. However, zinc did not induce any cytotoxic effect on both promyelocytic leukemic HL-60 cells and monoblastoid U937 cells. The nuclear morphology of Zn(2+)-treated H9c2 cells displayed apparent chromatin condensation, but no formation of chromatin fragmentation. In addition, phosphatidylserine (PS) externalization was observed by annexin-V staining. Zinc markedly decreased the intracellular GSH level in a time-dependent manner. Exposure to 0.2 mM ZnCl(2) for 6 hr decreased the intracellular GSH content to 13% of control value. Zinc-induced death of H9c2 cells and the intracellular GSH depletion were completely prevented by the addition of exogenous GSH and NAC. These result suggests that intracellular GSH depletion is directly involved in zinc-induced cardiomyopathy.
Animals ; Cardiomyopathies ; Cell Death* ; Cell Line ; Chromatin ; Doxorubicin ; Free Radicals ; HL-60 Cells ; Humans ; Rats ; U937 Cells ; Zinc

Purpose: The purpose of this pilot study is to evaluate the reliability and validity of the Korean version of the oral health impact profile (OHIP-EDENT K) for edentulous patients. Materials and Methods: The study was conducted on 12 patients who fabricated overdenture in the Department of Prosthodontics, Korea University, Guro Hospital. All subjects completed the Korean version of Oral Health Impact Profile (OHIP K) questionnaire. Shorten version of the OHIP called OHIP-14 K and OHIP-EDENT K were derived from the datasets. Cronbach's alpha was used to measure internal consistency of the summary scores for OHIP-EDENT K. The Spearman's correlation coefficient between the summary scores for OHIP-EDENT K and OHIP K was calculated to evaluate concurrent validity. Results: The reliability of the summary scores for OHIP-EDENT K was acceptable (α=.736). The Spearman's correlation coefficient of the summary scores for OHIP-EDENT K and OHIP K was 0.966, which was statistically significant (P<.001). OHIP-EDENT K exhibited less susceptibility to floor effects than OHIP-14 K and appeared to measure change as effectively as OHIP K. In order to prove the reliability, responsiveness and validity of OHIP-EDENT K, further studies with more samples are needed. Conclusion The OHIP-EDENT K, a questionnaire on oral health-related QOL comprising 19 items, has measurement properties comparable with the full 49-item version. This modified shortened version can be an alternative questionnaire to full version of OHIP K and OHIP-14 K in edentulous patients.

Purpose: The purpose of this pilot study is to evaluate the reliability and validity of the Korean version of the oral health impact profile (OHIP-EDENT K) for edentulous patients. Materials and Methods: The study was conducted on 12 patients who fabricated overdenture in the Department of Prosthodontics, Korea University, Guro Hospital. All subjects completed the Korean version of Oral Health Impact Profile (OHIP K) questionnaire. Shorten version of the OHIP called OHIP-14 K and OHIP-EDENT K were derived from the datasets. Cronbach's alpha was used to measure internal consistency of the summary scores for OHIP-EDENT K. The Spearman's correlation coefficient between the summary scores for OHIP-EDENT K and OHIP K was calculated to evaluate concurrent validity. Results: The reliability of the summary scores for OHIP-EDENT K was acceptable (α=.736). The Spearman's correlation coefficient of the summary scores for OHIP-EDENT K and OHIP K was 0.966, which was statistically significant (P<.001). OHIP-EDENT K exhibited less susceptibility to floor effects than OHIP-14 K and appeared to measure change as effectively as OHIP K. In order to prove the reliability, responsiveness and validity of OHIP-EDENT K, further studies with more samples are needed. Conclusion The OHIP-EDENT K, a questionnaire on oral health-related QOL comprising 19 items, has measurement properties comparable with the full 49-item version. This modified shortened version can be an alternative questionnaire to full version of OHIP K and OHIP-14 K in edentulous patients.

PURPOSE: To examine the feasibility of endoscopic thyroidectomy (ET) via an axillo-breast approach without gas insufflation for large thyroid tumors and micropapillary carcinomas. MATERIALS AND METHODS: The patients in the benign group were separated into groups 1 (n=95, <4 cm in tumor diameter) and 2 (n=37, > or =4 cm in tumor diameter). Also, 57 patients in the micropapillary carcinoma group underwent an endoscopic hemithyroidectomy (HT) (group 3) and were compared with 60 patients who received conventional open HT (group 4). Postoperative functional outcome, local complications, surgical outcomes, and pathological outcomes were compared between the groups. RESULTS: In the benign group, there was no significant difference in mean operating time, hospital stay, or overall perioperative complications between the two groups. In the micropapillary carcinoma group, mean operating time and hospital stay in group 3 were significantly longer than in group 4 (p=0.015 and p< or =0.001). The overall perioperative complications did not differ significantly between the groups. The postoperative cosmetic result was better in groups 1-3 (endo group) than in group 4 (open group). CONCLUSION: ET via a gasless axillo-breast approach seems to be a safe procedure even for benign thyroid lesions > or =4 cm and micropapillary carcinomas. Although it has the advantage of better cosmetic results over open thyroidectomy, there is room for improvement in terms of lessening its invasiveness and shortening the operative time.
Adult ; Carcinoma, Papillary/pathology/*surgery ; Endoscopy/adverse effects/methods ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Thyroid Nodule/pathology/*surgery ; Thyroidectomy/adverse effects/*methods ; Treatment Outcome

BACKGROUND: Idiopathic pulmonary fibrosis is a common interstitial lung disease; it is a chronic, progressive, and fatal lung disease of unknown etiology. Over the last two decades, knowledge about the underlying mechanisms of pulmonary fibrosis has improved markedly and facilitated the identification of potential targets for novel therapies. However, despite the large number of antifibrotic drugs being described in experimental pre-clinical studies, the translation of these findings into clinical practices has not been accomplished yet. NADH:quinone oxidoreductase 1 (NQO1) is a homodimeric enzyme that catalyzes the oxidation of NADH to NAD+ by various quinones and thereby elevates the intracellular NAD⁺ levels. In this study, we examined the effect of increase in cellular NAD⁺ levels on bleomycin-induced lung fibrosis in mice. METHODS: C57BL/6 mice were treated with intratracheal instillation of bleomycin. The mice were orally administered with β-lapachone from 3 days before exposure to bleomycin to 1-3 weeks after exposure to bleomycin. Bronchoalveolar lavage fluid (BALF) was collected for analyzing the infiltration of immune cells. In vitro, A549 cells were treated with transforming growth factor β1 (TGF-β1) and β-lapachone to analyze the extracellular matrix (ECM) and epithelial-mesenchymal transition (EMT). RESULTS: β-Lapachone strongly attenuated bleomycin-induced lung inflammation and fibrosis, characterized by histological staining, infiltrated immune cells in BALF, inflammatory cytokines, fibrotic score, and TGF-β1, α-smooth muscle actin accumulation. In addition, β-lapachone showed a protective role in TGF-β1–induced ECM expression and EMT in A549 cells. CONCLUSION: Our results suggest that β-lapachone can protect against bleomycin-induced lung inflammation and fibrosis in mice and TGF-β1–induced EMT in vitro, by elevating the NAD+/NADH ratio through NQO1 activation.
Actins ; Animals ; Bleomycin ; Bronchoalveolar Lavage Fluid ; Cytokines ; Epithelial-Mesenchymal Transition ; Extracellular Matrix ; Fibrosis* ; Idiopathic Pulmonary Fibrosis ; In Vitro Techniques ; Inflammation ; Lung Diseases ; Lung Diseases, Interstitial ; Lung* ; Mice* ; NAD ; Pneumonia ; Pulmonary Fibrosis ; Quinones ; Transforming Growth Factor beta1 ; Transforming Growth Factors