• Tuberculosis and Respiratory Diseases
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• v.56 no.2
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• pp.178-186
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• 2004

Background : Photodynamic therapy (PDT) is a new therapeutic method aimed at the selective destruction of cancer cells. The outcome is death of cancer cells through apoptosis or necrosis. The aim of this study was to investigate the characterization of PDT induced cell death in A549 lung cancer cells. Materials and methods : A549 cells were used as the lung cancer cell. 5 aminolevulinic acid (ALA) was used as the photosensitizer and a 632nm diode laser (Biolitec, Germany) as the light source. Cells were incubated with various concentrations of ALA. The 632nm diode laser was then administered for various laser irradiation times. The treated cells were incubated with 24, 48 and 72 hours. The cell viabilities were measured using the crystal violet assay and light microscopy. To observe the cell death mechanism after PDT, cells were observed under fluorescence microscopy after double staining with Hoechst 33342 and propium iodide after PDT. Results : In the crystal violet assay at 24 hours after PDT with a $3.2J/cm^2$ laser irradiation power, the cell viabilities were $89.56{\pm}4.11$, $87.67{\pm}5.48$, and $69.37{\pm}8.84$ with ALA concentrations of 10, 100, and $1mg/m{\ell}$, respectively. In crystal violet assay at 24 hours after PDT with $1mg/m{\ell}$ of ALA, the cell viabilities were $74{\pm}19.85$, $55{\pm}6.1$, and $49.06{\pm}16.64%$ with 1.6, 3.2 and $6.4J/cm^2$ laser irradiation powers, respectively. However, increasing the interval time after PDT did not change the cell viabilities. In the apoptosis assay, photodynamic therapy was inducing the apoptotic cell death. Conclusions : This study shows the apoptotic anticancer effect of photodynamic therapy in A549 lung cancer cells. However, further evaluations with other cancer cells and photosensitizers are necessary.

• Tuberculosis and Respiratory Diseases
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• v.64 no.4
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• pp.278-284
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• 2008

Background: TRAIL is a promising anticancer agent which induces selective tumor cell death due to a unique receptor system that includes death receptors and decoy receptors. DR5 TRAIL receptor is an originally identified p53-regulated death receptor gene that was induced, by doxorubicine, only in cells with a wild-type p53 status. We investigated that focused on the correlation between the DR5 and p53 expressions in non-small cell lung cancer (NSCLC). Methods: Immunohistochemical analysis, with using avidin-biotinylated horseradish peroxidase complex, was carried out in 89 surgically resected NSCLC formalin-fixed paraffin-embedded tissue sections. As primary antibodies, we used anti-DR5 polyclonal antibody and anti-p53 monoclonal antibody. A negative control was processed with each slide. The positive tumor cells were quantified twice and these values were expressed as percentage of the total number of tumor cells, and the intensity of immunostaining was expressed. The analysis of the DR5 expression was done separately in tumor area and in a nearby region of normal tissue. Results: The DR5 expression was high in the bronchial epithelium (89% of cases) but this was almost absent in type I & II pneumocytes, lymphocytes and smooth muscle cells. High DR5 expression rate in tumor was seen in 28% (15/53) of squamous cell carcinomas, in 47% (15/32) of adenocarcinomas and, in 50% (2/4) of large cell carcinomas. The DR5 expression did not show any statistical significance relationship with the T stage, N stage, or survival. However, the DR5 expression showed significant inverse correlation with the p53 expression. (p< 0.01). Conclusion: We demonstrated that the DR5 expression in NSCLC via immunohistochemical analysis is relatively tumor-specific except for that in the normal bronchial epithelium and it is significantly dependent on the p53 status. This might be in vivo evidence for the significance of the DR5 gene as a p53 downstream gene.

• Tuberculosis and Respiratory Diseases
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• v.65 no.6
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• pp.476-486
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• 2008

Background: TRAIL (TNF-related apoptosis inducing ligand) is a newly identified member of the TNF gene family which appears to have tumor-selective cytotoxicity due to the distinct decoy receptor system. TRAIL has direct access to caspase machinery and induces apoptosis regardless of p53 phenotype. Therefore, TRAIL has a therapeutic potential in lung cancer which frequently harbors p53 mutation in more than 50% of cases. However, it was shown that TRAIL also could activates $NF-{\kappa}B$ in some cell lines which might inhibit TRAIL-induced apoptosis. This study was designed to investigate whether TRAIL can activate $NF-{\kappa}B$ in lung cancer cell lines relatively resistant to TRAIL-induced apoptosis and inhibition of $NF-{\kappa}B$ activation using proteasome inhibitor MG132 which blocks $I{\kappa}B{\alpha}$ degradation can sensitize lung cancer cells to TRAIL-induced apoptosis. Methods: A549 (wt p53) and NCI-H1299 (null p53) lung cancer cells were used and cell viability test was done by MTT assay. Apoptosis was confirmed with Annexin V assay followed by FACS analysis. To study $NF-{\kappa}B$-dependent transcriptional activation, a luciferase reporter gene assay was used after making A549 and NCI-H1299 cells stably transfected with IgG ${\kappa}-NF-{\kappa}B$ luciferase construct. To investigate DNA binding of $NF-{\kappa}B$ activated by TRAIL, electromobility shift assay was used and supershift assay was done using anti-p65 antibody. Western blot was done for the study of $I{\kappa}B{\alpha}$ degradation. Results: A549 and NCI-H1299 cells were relatively resistant to TRAIL-induced apoptosis showing only 20~30% cell death even at the concentration 100 ng/ml, but MG132 ($3{\mu}M$) pre-treatment 1 hour prior to TRAIL addition greatly increased cell death more than 80%. Luciferase assay showed TRAIL-induced $NF-{\kappa}B$ transcriptional activity in both cell lines. Electromobility shift assay demonstrated DNA binding complex of $NF-{\kappa}B$ activated by TRAIL and supershift with p65 antibody. $I{\kappa}B{\alpha}$ degradation was proven by western blot. MG132 completely blocked both TRAIL-induced $NF-{\kappa}B$ dependent luciferase activity and DNA binding of $NF-{\kappa}B$. Conclusion: This results suggest that inhibition of $NF-{\kappa}B$ can be a potentially useful strategy to enhance TRAIL-induced tumor cell killing in lung cancer.

• Journal of the Korean earth science society
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• v.36 no.7
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• pp.661-673
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• 2015

UBVI CCD photometry of the intermediate age open cluster NGC 7790 has been obtained using AZT-22 1.5 m telescope (f/7.74) at the Maidanak Astronomical Observatory in Uzbekistan. NGC 7790 contains three ${\delta}$ Cep variable stars including CEa Cas, CEb Cas, and CF Cas. PSF photometry was carried out using IRAF/DAOPHOT for all observations. The total number of stars observed both in V and I filter was 1008 and the limiting magnitude was $V{\approx}22$. To determine atmospheric extinction coefficients and photometric zero points, many blue and red standard stars as well as the standard stars in the celestial equator under various airmass were observed. Photometric data were transformed into the standard Johnson-Cousins' UBVI standard system. From the analysis of UBVI color-magnitude diagram and color-color diagram, the color excess in V and I filter [$E(B-V)=0.58{\pm}0.02$], the selective extinction ratio in V and I filter [$R_V{\equiv}A_V/E(B-V)=3.02{\pm}0.09$] and distance modulus ($V_0-M_V=12.65{\pm}0.10$) of the cluster were determined. The age of the cluster was estimated to be log $age=8.05{\pm}0.05$ [yr] based on the position of these three Cepheid variables in the color-magnitude diagram, the isochrone of the Geneva group ($Ekstr{\ddot{o}}m$ et al., 2012-Z=0.019), and the isochrone of the Padova group (Bressan et al., 2012-Z=0.014) were used to compare each other. Of them, the Geneva models that considered stellar rotation well described the position of ${\delta}$ Cepheid variables in the blue loop. Although they were well consistent with standard period-luminosity relation of ${\delta}$ Cepheid variables, three Cepheid variables in NGC 7790 were, on average, brighter by about 0.5 mag than the absolute magnitude estimated from the mean period-luminosity relation at a given period.

• Journal of Aquaculture
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• v.20 no.4
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• pp.278-288
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• 2007

Shrimp culture in Korea had been rapidly developed during 1990's and the production of farmed shrimp reached 3,268 mt from 2,605 ha in 2001. However the shrimp production decreased to 2,368 mt in 2004 because of the mass mortality due to outbreak of white spot syndrome virus (WSSV). WSSV which is one of the most serious threats associated with cultured shrimp around the world has given the economic damages to shrimp culture industry every year since it was found from the shrimp ponds in the west coast of the South Korea in 1993. Various polyculture technologies of shrimp with shellfish, finfish or seaweeds have been implemented to reduce economic damages by mass mortalities of shrimp. Among them, the polyculture of shrimp with carnivorous fish can suppress or delay the viral outbreak of shrimp ponds because the fish may selectively eat the moribund shrimps infected by virus. To determine the selective predatory effect of river puffer Takifugu obscures on WSSV infected shrimp, postlarvae of Litopenaeus vannamei and Fenneropenaeus chinensis. One-year old river puffers were stocked to four earthen ponds of $1,616-1,848\;m^2$ in surface area as followings: polyculture LvP, L. vannamei ($43.4/m^2$)+puffer ($0.22/m^2$); control Lv, L. vannamei ($46.9/m^2$); polyculture FcP, F. chinensis ($30.3/m^2$)+puffer ($0.25/m^2$); control Fc, F. chinensis ($24.6/m^2$). Ponds of control Fc and polyculture FcP had mass mortalities by WSSV outbreak on the $51^{st}$ and $57^{th}$ days of culture respectively. The shrimps of polyculture LvP and control Lv were harvested on the $95^{th}\;day$. Shrimp survival rates of polyculture LvP and control Lv were 32.4% and 18.2% respectively and shrimp productivity of polyculture LvP was 69.2% higher than that of control Lv. Concentration of nutrients (TAN, $NO_2-N$, $NO_3-N$) was maintained within optimal ranges for shrimp growth although that of polyculture ponds showed at least two times higher than that of control ponds. The results suggest that polyculture of L. vannamei with river puffer is higher than monoculture in survival rate and productivity. In addition, F. chinensis should be carefully cultured because this species shows much higher susceptibility to WSSV than L. vannamei.

• Sleep Medicine and Psychophysiology
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• v.12 no.2
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• pp.122-132
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• 2005

Objectives: The objective of this study is to assess cognitive functions and their relationship with sleep symptoms in young narcoleptic patients. Methods: Eighteen young narcolepsy patients and 18 normal controls (age: 17-35 years old) were recruited. All narcolepsy patients had HLA $DQB_1$ *0602 allele and cataplexy. Several important areas of cognition were assessed by a battery of neuropsychological tests consisting of 13 tests: executive functions (e.g. cognitive set shifting, inhibition, and selective attention) through Wisconsin card sorting test, Trail Making A/B, Stroop test, Ruff test, Digit Symbol, Controlled Oral Word Association and Boston Naming Test; alertness and sustained attention through paced auditory serial addition test; verbal/nonverbal short-term memory and working memory through Digit Span and Spatial Span; visuospatial memory through Rey-Osterrieth complex figure test; verbal learning and memory through California verbal learning test; and fine motor activity through grooved pegboard test. Sleep symptoms in narcolepsy patients were assessed with Epworth sleepiness scale, Ullanlinna narcolepsy scale, multiple sleep latency test, and nocturnal polysomnography. Relationship between cognitive functions and sleep symptoms in narcolepsy patients was also explored. Results: Compared with normal controls, narcolepsy patients showed poor performance in paced auditory serial addition (2.0 s and 2.4 s), digit symbol tests, and spatial span (forward)(t=3.86, p<0.01; t=-2.47, p=0.02; t=-3.95, p<0.01; t=-2.22, p=0.03, respectively). There were no significant between-group differences in other neuropsychological tests. In addition, results of neuropsychological test in narcolepsy patients were not correlated with Epworth sleepiness scale score, Ullanlinna narcolepsy scale score and sleep variables in multiple sleep latency test or nocturnal polysomnography. Conclusion: The current findings suggest that young narcolepsy patients have impaired attention. In addition, impairment of attention in narcolepsy might not be solely due to sleep symptoms such as excessive daytime sleepiness.

• Nuclear Medicine and Molecular Imaging
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• v.40 no.6
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• pp.322-326
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• 2006

A low iodine diet (LID) for $1{\sim}2$ weeks is recommended for patients who undergoing radioiodine remnant ablation. However, the LID educations for patients are different among centers because there is no concrete recommendation for protocol of LID. In this investigation, we compared two representative types of LID protocols performed in several centers in Korea using urine iodine to creatinine ratio (urine I/Cr). Methods: From 2006, April to June, patients referred to our center for radioiodine remnant ablation of thyroid cancer from several local hospitals which had different LID protocols were included. We divided into two groups, stringent LID for 1week and less stringent LID for 2 weeks, then measured their urine I/Cr ratio with spot urine when patients were admitted to the hospital. Results: Total 27 patients were included in this investigation (M:F=1:26; 13 in one-week stringent LID; 14 in two-week less stringent LID. Average of urine I/Cr ratio was $127.87{\pm}78.52{\mu}g/g$ in stringent LID for 1 week, and $289.75{\pm}188.24{\mu}g/g$ in less stringent LID for 2 weeks. It was significantly lower in stringent LID for 1 week group (p=0.008). The number of patients whose urine I/Cr ratios were below $100{\mu}g/g$ was 6 of 13 in stringent LID for 1 week group, and 3 of 14 in less stringent LID for 2 weeks group. Conclusion: Stringent LID for 1 week resulted in better urinary I/Cr ratio in our investigation compared with the other protocol. However it still resulted in plenty of inadequate range of I/Cr ratio, so more stringent protocol such as stringent LID for 2 weeks is expected more desirable.

• Clinical and Experimental Pediatrics
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• v.51 no.8
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• pp.879-885
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• 2008

Purpose : The human lung fibroblast may act as an immunomodulatory cell by providing pro-inflammatory cytokines and chemokines, which are important in airway remodeling. Vascular endothelial growth factor (VEGF) induces mucosal edema and angiogenesis. Thymus and activation regulated chemokine (TARC) induces selective migration of T helper 2 cells. We investigated whether human lung fibroblasts produced VEGF and TARC, and the effects were augmented with the co-culture of fibroblasts and human bronchial smooth muscle cells (HBSMC), and whether dexamethasone can inhibit the proliferation and the release of VEGF in lung fibroblasts. Methods : Human lung fibroblasts were cultured with and without HBSMC, growth-arrested in serum-deprived medium, and pretreated with dexamethasone for 16 hours. After 24-hour stimulation with platelet derived growth factor-BB (PDGF-BB) and/or transforming growth factor-${\beta}$ (TGF-${\beta}$), culture supernatant was harvested for assays of VEGF and TARC. Cell proliferation was assayed using BrdU cell proliferation ELISA kit. Results : 1) The release of VEGF was significantly increased after stimulation with TGF-${\beta}$, and its release was augmented when co-stimulated with PDGF and TGF-${\beta}$. 2) VEGF release induced by PDGF or TGF-${\beta}$ was inhibited by dexamethasone. 3) There was no synergistic effect on the release of VEGF when human lung fibroblasts were co-cultured with HBSMC. 4) Dexamethasone did not suppress human lung fibroblasts proliferations. 5) Neither TGF-${\beta}$ nor PDGF induced TARC release from lung fibroblasts. Conclusion : Human lung fibroblasts may modulate airway remodeling by release of VEGF, but they have no synergistic effects when co-cultured with HBSMC. Dexamethasone suppresses VEGF release, not proliferation of lung fibroblast.

• The korean journal of orthodontics
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• v.33 no.3 s.98
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• pp.169-183
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• 2003

Tooth movement by orthodontic force effects great tissue changes within the periodontium, especially by shifting the blood flow in the pressure side and resulting in a hypoxic state of low oxygen tension. The aim of this study is to elucidate the possible mechanism of apoptosis in response to hypoxia in MC3T3El osteoblasts, the main cells in bone remodeling during orthodontic tooth movement. MC3T3El osteoblasts under hypoxic conditions ($2\%$ orygen) resulted in apoptosis in a time-dependent manner as estimated by DNA fragmentation assay and nuclear morphology stained with fluorescent dye, Hoechst 33258. Pretreatment with Z-VAD-FMK, a pancaspase inhibitor, or Z-DEVD-CHO, a specific caspase-3 inhibitor, completely suppressed the DNA ladder in response to hypoxia. An increase in caspase-3-like protease (DEVDase) activity was observed during apoptosis, but no caspase-1 activity (YVADase) was detected. To confirm what caspases are involved in apoptosis, Western blot analysis was performed using anti-caspase-3 or -6 antibodies. The 10-kDa protein, corresponding to the active products of caspase-3, and the 10-kDa protein of the active protein of caspase-6 were generated in hypoxia-challenged cells in which the processing of the full length form of caspase-3 and -6 was evident. While a time course similar to this caspase-3 and -6 activation was evident, hypoxic stress caused the cleavage of lamin A, which was typical of caspase-6 activity. In addition, the stress elicited the release of cytochrome c into the cytosol during apoptosis. Furthermore, we observed that pre-treatment with SB203580, a selective p38 mitogen activated protein kinase inhibitor, attenuated the hypoxia-induced apoptosis. The addition of SB203S80 suppressed caspase-3 and -6-like protease activity by hypoxia up to $50\%$. In contrast, PD98059 had no effect on the hypoxia-induced apoptosis. To confirm the involvement of MAP kinase, JNK/SAPK, ERK, or p38 kinase assay was performed. Although p38 MAPK was activated in response to hypoxic treatment, the other MAPK -JNK/SAPK or ERK- was either only modestly activated or not at all. These results suggest that p38 MAPK is involved in hypoxia-induced apoptosis in MC3T3El osteoblasts.

• Journal of Oral Medicine and Pain
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• v.32 no.4
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• pp.375-381
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• 2007

Occlusal stabilization appliance is one of the most common treatment option for management of temporomandibular disorders. It acts in oral cavity for several hours per day, and usually it will take at least 6 months to 2 years of total wearing periods to take a treatment goal. In the oral cavity, occlusal stabilization appliance, unintentional manner, is able to acts as a reservoir of bacteria and protect bacteria from saliva and oxygen. This condition is so favorable to many bacteria such as S. mutans and other anaerobes, usually have been reported as causative factors of dental caries, periodontal disease and oral malodor. In this study, we investigated anaerobic bacteria and S. mutans count before and after occlusal stabilization appliance use to evaluate the possible role of occlusal stabilization appliance as protector of these bacteria. Four men(average 27.5 years) wore maxillary occlusal stabilization appliance at each night(average 9 hours) for 5 days. we swabbed saliva-plaque mixed sample at 3 different site(maxillary left 2nd molar, maxillary left central incisor, mandibular left 2nd molar) before and after occlusal stabilization appliance use. Each samples were plated in (1) anaerobic blood agar medium, (2) selective S. mutans medium(MS-MUTV) and incubated in anaerobic chamber($CO^2$ 10%, $37^{\circ}C$) for 72 hours. Each bacterial colony forming unit(CFU) were counted with naked eyes. From obtained data, we can conclude as follows: 1. There was some changes about anaerobic bacteria and S. mutans count in oral cavity after occlusal stabilization appliance use. 2. The number of anaerobic bacteria was significantly increased at maxillary 2nd molar(P=0.003), maxillary central incisor(P=0.020) after occlusal stabilization appliance use compared with before. 3. Occlusal stabilization appliance use itself had indirect effect to increase the number of anaerobic bacteria at other uncovered opponent tooth site. 4. The number of S. mutans was significantly increased at maxillary 2nd molar(P=0.043), maxillary central incisor (P=0.049) after occlusal stabilization appliance use compared with before. 5. Occlusal stabilization appliance use itself had not any effect on the number of S. mutans at other uncovered opponent tooth site.