A bacterial strain, producing an excellent lipolytic enzyme, was isolated from the intestinal tracts of an earthworm (Eisenia fetida). The strain was identified as Aeromonas hydrophila by phenotypic, chemotaxonomic characteristics and 16S ribosomal DNA analysis, and was designated as Aeromona hydrophila PL43. The lipolytic enzyme from A. hydrophila PL43 was purified via 35−45% ammonium sulfate precipitation, DEAE-sepharose fast flow ion-exchange, and sephacryl S-300HR gel filtration chromatography. The yield of the purified enzyme was 3.7% and 2.5% of the total activity of crude extracts with p-nitrophenyl butyrate (pNPB) and p-nitrophenyl palmitate (pNPP) as substrates, respectively. The molecular weight of the purified enzyme was approximately 74 kDa using gel filtration, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and zymography. The optimal activity of purified enzyme was observed at 50℃ and pH 8.0 using pNPB, and 60℃ and pH 8.0 using pNPP. The purified enzyme was stable in the ranges 20− 60℃ and pH 7.0−10.0. The activity of purified enzyme was inhibited by PMSF, pepstatin A, Co2+, Cu2+, and Fe2+, but was recovered by metal chelating of EDTA. The Km and Vmax values of the purified enzyme were 1.07 mM and 7.27 mM/min using pNPB and 1.43 mM and 2.72 mM/min using pNPP, respectively.
Daejeon-sa basalt in the Mt. Juwang area composed of 12 basalt flows alternate with 9 peperites and each basalt and peperite has the variety of thickness. Peperites yielded in Daejeon-sa basalt are mixed of basalt with reddish shale, of which textural type is globular peperite. Basalts yielded in Daejeon-sa basalt are massive basalt without vesicule, although sometimes vesicules are founded in upper within a flow unit. The basalt has mainly pseudomorph of olivine as phenocryst, and also plagioclase and clinopyroxene phenocryst. Matrix is mainly subophitic texture. The plotting result on the TAS diagram shows these basalts belong to the sub-alkaline, and it can be subdivided into calc-alkaline series on the basis of the diagram of Si02 vs. K20 and of alkali index vs. A1203 diagram. According to plots of wt.% oxides vs. wt.% MgO, abundances of A1203 and CaO increase with decreasing MgO while F ~ dOecre~ase . With decreasing MgO compatible elements decrease while incompatible elements increase. In spider diagram of MORB-normalized trace element patterns, HFS elements are nearly similiar with MORB, but LIL elements are enriched. Especially, contents of Ce, F: and Sm are enriched but Nb is depleted. In the chondrite-normalized REE patterns light REEs are enriched than heavy REEs. Tectomagmatic discrimination diagrams shows basalts in the study area are formed in the tectonomagmatic environment of subduction zone under continental margin. This result accord with characters of chemical composition mentioned above. Cr vs. Y diagram and CeM, vs. Ce diagram show that the primary magma of the basalts may formed by the about 15% partial melting of garnet-peridotite in the mantle wedge. After then, Daejeon-sa basalts may formed from evolved magma undergone mainly olivine fractional crystallization and contarnination of crustal materials before eruption.
Typha orientalis, also known as bulrush or cattail, is a perennial herbaceous plant found in freshwater wetlands and has been widely used in constructed wetlands for wastewater treatment. Recent data has revealed that SH21B, a mixture composed of seven herbs including T. orientalis, exhibited an anti-adipogenic activity by the inhibition of the expression of adipogenic regulators. However, the anti-cancer effect of T. orientalis and its molecular mechanisms remain unclear. In this study, we evaluated the anti-cancer effect and its mechanism in the methanol extract of T. orientalis (METO) on human colon carcinoma HT29 cells. It was found that METO treatment showed cytotoxic activity in a dose-dependent manner, and induced G2/M cell cycle arrest and apoptosis in HT29 cells. The induction of G2/M arrest by METO was associated with the up-regulation of phospho-Cdc2 (Tyr15), an inactive form of Cdc2 and the down-regulation of Cdc25c phosphatase. METO also induced tumor suppressor p53 and cyclin-dependent kinase inhibitor p21 (WAF1/CIP1) expression. In addition, METO-induced apoptosis was characterized by the proteolytic activation of caspase-3, degradation of poly ADP ribose polymerase (PARP), and up-regulation of death receptor FAS and pro-apoptotic Bax expression. Collectively, these results indicate that the cell cycle inhibition and apoptosis induction of METO in HT29 cells allows for the possibility of its use in anti-cancer therapies.
Background : Heme oxygenase-1 (HO-1) is an inducible enzyme that catalyzes the oxidative degradation of heme to form biliverdin, carbon monoxide (CO), and free iron. The current evidence has indicated a critical role of HO-1 in cytoprotection and also in other, more diverse biological functions. It is known that the high expression of HO-1 occurs in various tumors, and that HO-1 has an important role in rapid tumor growth because of its antioxidative and antiapoptotic effects. Therefore, the role of HO-1 was analyzed in human lung cancer cell lines, and especially in the A549 cell line. Material and Methods : Human lung cancer cell lines, i.e., A549, NCI-H23, NCI-H157 and NCI-H460, were used for this study. The expression of HO-1 in the untreated state was defined by Western blotting. ZnPP, which is the specific HO inhibitor we used, and the viability of cells were tested for by conducting MTT assaysy. The HO enzymatic activity, as determined via the bilirubin level, was also indirectly measured. Moreover, the generation of intracellular hydrogen peroxide (H2O2) was monitored fluorimetrically with using a scopoletin-horse radish peroxidase (HRP) assay and 2',7'-dichlorofluorescein diacetate (DCFH-DA). We have also transfected small HO-1 interfering RNA (siRNA) into A549 cells, and the apoptotic effects were evaluated by flow cytometric analysis and Western blotting. Results : The A549 cells had a greater expression of HO-1 than the other cell lines, whereas ZnPP significantly decreased the viability of the A549 cells more than the viability of the other lung cancer cells in a dose-dependant fashion. Consistent with the viability, the HO enzymatic activity also was decreased. Moreover, intracellular H2O2 generation via ZnPP was induced in a dose-dependent manner. Apoptotic events were, then induced in the HO-1 siRNA transfected A549 cells. Conclusion : HO-1 provides new important insights into the possible molecular mechanism of the antitumor therapy in lung cancer.
Wetland plants have evolved specialized adaptations to survive in the low-oxygen conditions associated with prolonged flooding. The development of internal gas space by means of aerenchyma is crucial for wetland plants to transport $O_2$ from the atmosphere into the roots and rhizome. The formation of tissue with high porosity depends on the species and environmental condition, which can control the depth of root penetration and the duration of root tolerance in the flooded sediments. The oxygen in the internal gas space of plants can be delivered from the atmosphere to the root and rhizome by both passive molecular diffusion and convective throughflow. The release of $O_2$ from the roots supplies oxygen demand for root respiration, microbial respiration, and chemical oxidation processes and stimulates aerobic decomposition of organic matter. Another essential mechanism of wetland plants is downward water movement across the root zone induced by water uptake. Natural and constructed wetlands sediments have low hydraulic conductivity due to the relatively fine particle sizes in the litter layer and, therefore, negligible water movement. Under such condition, the water uptake by wetland plants creates a water potential difference in the rhizosphere which acts as a driving force to draw water and dissolved solutes into the sediments. A large number of anatomical, morphological and physiological studies have been conducted to investigate the specialized adaptations of wetland plants that enable them to tolerate water saturated environment and to support their biochemical activities. Despite this, there is little knowledge regarding how the combined effects of wetland plants influence the biogeochemistry of wetland sediments. A further investigation of how the Presence of plants and their growth cycle affects the biogeochemistry of sediments will be of particular importance to understand the role of wetland in the ecological environment.
Journal of the Korean Society of Marine Environment & Safety
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v.18
no.5
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pp.398-405
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2012
PAHs(Polycyclic Aromatic Hydrocarbons) in coastal area and estuary adjacent to urban area and industrial activities region are mainly introduced into marine environment via atmosphere and river. This study which is evaluated the distribution characteristics of PAHs discharge from rivers and sewage disposal water which flowing into Ulsan bay, and were carried out in Jun of dry season and in Auguest of wet season, 2008. The water samples from eight main rivers and Youngyeon WWTP(Wastewater Treatment Plant) flowed into Ulsan bay were taken for analysis of dissolved PAHs. The range and mean values of dissolved PAHs concentrations showed 10.30 to 87.88 ng/L, its mean 36.94 ng/L in dry season and 10.30 to 69.57 ng/L, its mean 24.37 ng/L in wet season. The distribution of dissolved PAHs showd the high concentrations in the Gungcheoncheon which is flowed from urban and industrial area. The ranges and means values of the loading fluxes were estimated with 0.04 to 8.27 g/day, its mean 2.05 g/day in dry season, and 0.03 to 4.77 g/day, its mean 1.61 g/day, in wet season. The loading flux showed the highest value in Taewha river due to the high flow rate and the urban activity. The composition patterns of dissolved PAHs compound showed most of the trend occupying low molecuar weight PAHs than high molecular weight PAHs. These results were due to physical and chemical characteristics of PAHs compound, and were similar to those of other studies. The concentrations of dissolved PAHs in this study are lower than those of other studies, and the level of PAHs pollution appeared to be not serious in reverine waters of Ulsan coast.
Objective: The present study employed 5-aza-2'-deoxycytidine (5-Aza-CdR) to treat non-small cell lung cancer (NSCLC) cell line A549 to investigate the effects on proliferation and expression of the TFPI-2 gene. Methods: Proliferation was assessed by MTT assay after A549 cells were treated with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR, a specific demethylating agent, for 24, 48 and 72h. At the last time point cells were also analyzed by flow cytometry (FCM) to identify any change in their cell cycle profiles. Methylation-specific polymerase chain reaction (MSPCR), real time polymerase chain reaction(real-time PCR) and western blotting were carried out to determine TFPI-2 gene methylation status, mRNA expression and protein expression. Results: MTT assay showed that the growth of A549 cells which were treated with 5-Aza-CdR was significantly suppressed as compared with the control group (0 ${\mu}mol/L$ 5-Aza-CdR). After treatment with 0, 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h, FCM showed their proportion in G0/G1 was $69.7{\pm}0.99%$, $76.1{\pm}0.83%$, $83.8{\pm}0.35%$, $95.5{\pm}0.55%$ respectively (P<0.05), and the proportion in S was $29.8{\pm}0.43%$, $23.7{\pm}0.96%$, $15.7{\pm}0.75%$, $1.73{\pm}0.45%$, respectively (P<0.05), suggesting 5-Aza-CdR treatment induced G0/G1 phase arrest. MSPCR showed that hypermethylation in the promoter region of TFPI-2 gene was detected in control group (0 ${\mu}mol/L$ 5-Aza-CdR), and demethylation appeared after treatment with 1, 5, 10 ${\mu}mol/L$ 5-Aza-CdR for 72h. Real-time PCR showed that the expression levels of TFPI-2 gene mRNA were $1{\pm}0$, $1.49{\pm}0.14$, $1.86{\pm}0.09$ and $5.80{\pm}0.15$ (P<0.05) respectively. Western blotting analysis showed the relative expression levels of TFPI-2 protein were $0.12{\pm}0.01$, $0.23{\pm}0.02$, $0.31{\pm}0.02$, $0.62{\pm}0.03$ (P<0.05). TFPI-2 protein expression in A549 cells was gradually increased significantly with increase in the 5-Aza-CdR concentration. Conclusions: TFPI-2 gene promoter methylation results in the loss of TFPI-2 mRNA and protein expression in the non-small cell lung cancer cell line A549, and 5-Aza-CdR treatment could induce the demethylation of TFPI-2 gene promoter and restore TFPI-2 gene expression. These findings provide theoretic evidence for clinical treatment of advanced non-small cell lung cancer with the demethylation agent 5-Aza-CdR. TFPI-2 may be one molecular marker for effective treatment of advanced non-small cell lung cancer with 5-Aza-CdR.
Kim, Jung-Hee;Song, Ho-Chun;Yang, Jong-Chul;Lee, Byeong-Il;Heo, Young-Jun;Bom, Hee-Seung;Park, Tae-Jin;Min, Jung-Joon
Nuclear Medicine and Molecular Imaging
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v.40
no.6
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pp.302-308
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2006
Purpose: Although several neuroanatomical models of panic disorder have been proposed, little is known regarding the neurological mechanisms underlying cognitive-behavioral therapy (CBT) in patients with panic disorder. This study was performed to identify the brain structures that show changes of regnioal cerebral blood flow (rCBF) after CBT in patients with panic disorder. Materials and Methods: Seven patients who were diagnosed as panic disorder by DSM-IV were treated with CBT for 8 weeks and twelve healthy volunteers joined in this study. Serial $^{99m}Tc-ECD$ brain perfusion SPECT images were acquisited and PDSS-SR (Self-Report version of Panic Disorder Severity Scale) and ACQ (Agoraphobic Cognitive Question) scores were measured just before and after CBT in all patients. Data were analyzed using SPM2. Results: Subjective symptoms were improved, and PDSS-SR and ACQ scores were significantly reduced ($14.9{\pm}3.9\;vs.\;7.0{\pm}1.8$, p<0.05; $30.3{\pm}8.5\;vs.\;21.6{\pm}3.4$, p<0.05, respectively) after CBT in panic patients. Before CBT, a significant increase of rCBF was found in the cingulate gylus, thalamus, midbrain, both medial frontal and temporal lobes of the panic patients compared to the normal volunteers. After CBT, we observed a significant rCBF decrease in the left parahippocamus, right insula and cingulate gyrus, both frontal and temporal lobes, and a significant rCBF increase in both the occipital lobes, left insula, both frontal and left parietal lobes. Conclusion: These data suggested that CBT is effective for panic disorder and diminish the activity of the brain areas associated with fear in panic disorder.
Park, Kyung-Won;Kang, Do-Young;Park, Min-Jeong;Cheon, Sang-Myung;Cha, Jae-Kwan;Kim, Sang-Ho;Kim, Jae-Woo
Nuclear Medicine and Molecular Imaging
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v.41
no.6
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pp.530-537
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2007
Purpose: The aim of this study is to assess the specific patterns of regional cerebral blood flow (rCBF) in patients with the early stage of subcortical vascular dementia (SVaD) and Alzheimer's disease (AD) using Tc-99m HMPAO SPECT, and to compare the differences between the two conditions. Materials and Methods: Sixteen SVaD, 46 AD and 12 control subjects participated in this study. We included the patients with SVaD and AD according to NINCDS-ADRDA and NINDS-AIREN criteria. They were all matched for age, education and clinical dementia rating scores. Three groups were evaluated by Tc-99m HMPAO SPECT using statistical parametric mapping (SPM) for measuring rCBF. The SPECT data of patients with SVaD and AD were compared with those of normal control subjects and then compared with each other. Results: SPM analysis of the SPECT image showed significant perfusion deficits on the right temporal region and thalamus, left insula and superior temporal gyrus, both cingulate gyri and frontal subgyri in patients with SVaD and on the left supramarginal gyrus, superior temporal gyrus, postcentral gyrus and inferior parietal lobule, right fugiform gyrus and both cingulate gyri in AD compared with control subjects (uncorrected p<0.01). SVaD patients revealed significant hypoperfusion in the right parahippocampal gyrus with cingulated gyrus, left insula and both frontal subgyral regions compared with AD (uncorrected p<0.01). Conclusion: Our study shows characteristic and different pattern of perfusion deficits in patients with SVaD and AD, and these results may be helpful to discriminate the two conditions in the early stage of illness.
Kim, Min-Jeong;Kim, Jihee;Lee, Jin-Ho;Kim, Minah;Woo, Keunjung;Kim, Han Sung;Kim, Tack-Joong
Journal of Life Science
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v.31
no.9
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pp.787-795
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2021
Eupatorium chinensis var. simplicifolium (EUC) has anti-inflammatory and antioxidant effects. Young sprouts of EUC have been used as food for a long time, and the whole EUC plant has been used as an herbal remedy in oriental medicine. Arteriosclerosis, or chronic inflammation in arterial vessels, is a cardiovascular disease and is involved in various disorders. Cardiovascular diseases such as restenosis and neuropathic hyperplasia are mainly caused by abnormal growth and movement due to multiple growth factors in vascular smooth muscle cells (VSMCs). Platelet-derived growth factor (PDGF) is a mitogen released from damaged vessel walls and is involved in the proliferation and migration of VSMCs. To determine the effects of EUC on the abnormal proliferation and migration of VSMCs, the present study investigated intracellular signaling pathways in PDGF-BB-induced VSMCs treated with and without EUC. Pretreating PDGF-BB-induced VSMCs with EUC tended to effectively decrease cell proliferation and migration. Subsequently, the intracellular growth-related signaling pathways of AKT, phospholipase C gamma (PLC-γ), and mitogen-activated protein kinase (MAPK) were investigated using western blotting to confirm inhibited phosphorylation. Furthermore, flow cytometry data showed that EUC blocked the cell cycle of VSMCs. These results suggest that EUC can inhibit the proliferation and migration of VSMCs by controlling the cell cycle and growth factor receptors. Furthermore, this indicates that EUC can be used as a preventative against cardiovascular disease resulting from abnormal proliferation and migration of VSMCs.
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