Hizikia fusiforme is a kind of brown edible seaweed that mainly grows in the temperate seaside areas of the northwest pacific, including Korea, Japan and China, and has been widely used as a health food for hundreds of years. Recently, H. fusiforme has been known to exert pharmacological activities including antioxidant, antimutagenic and anticoagulant activities. However, the molecular mechanisms of H. fusiforme in malignant cells have not been clearly elucidated yet. In this study, the effects of ethyl alcohol extract of H. fusiforme (EAHF) on the anti-proliferative effects of MDA-MB-231 and MCF-7 human breast cancer cells were investigated. EAHF treatment resulted in a concentration-dependent growth inhibition by including apoptosis in MDA-MB-231 cells and G1 phase arrest in MCF-7 cells, which could be proved by MTT assay, DAPI staining, agarose gel electrophoresis and flow cytometry analysis. In MDA-MB-231 cells, the increase in apoptosis induced by EAHF treatment correlated with up-regulation of pro-apoptotic Bax expression. EAHF treatment induced the proteolytic activation of caspase-3 and caspase-9, and a concomitant inhibition of poly (ADP-ribose) polymerase, $\beta$-catenin, phospholipase-${\gamma}1$ protein and DNA fragmentation factor 45/inhibitor of caspase-activated DNase. Taken together, these findings provide important new insights into the possible molecular mechanisms of the anti-cancer activity of H. fusiforme.
This study analyzed nonmetallic inclusions in iron swords with a ring pommel excavated in the Ipbuk-dong, Suwon. Scanning electron microscopy with energy dispersive spectroscopy(SEM-EDS) was used to estimate the iron-making temperature, and we compared the oxide with $SiO_2$ to investigate the heat-treatment technology in the production of iron swords with a ring pommel by investigating the artificial insertion of a slag former and the metallurgical structure. From the wustite observed in most of the specimens, it is judged that these swords were produced by heating and forging iron smelted at a low temperature using the solid reduction method. In addition, judging from the partial presence of $P_2O_5$, it is assumed that they were smelted directly with natural ore, not calcined. From the ratios of $CaO/SiO_2$ and $TiO_2/SiO_2$, it is judged that the raw material for iron-making was iron ore and that a calcareous slag former was not artificially inserted. The structure of the blade part on the front end was pure iron. From the high carbon content of the blade part on the ring pommel and the formation of a martensitic structure and pearlite colony, it is judged that they were tempered after carburizing and that the back, handle part, and ring pommel were unintentionally carburized. Judging from the structure of these specimens, it was noted that they were produced by applying artificial partial heat-treatment technology. This study attempted to present a more scientific analysis by using the method of interpretation through component analysis of nonmetallic inclusions appearing in one relic by the ratio of the oxide divided by $SiO_2$. It is judged that reinterpreting the arguments by the results of the existing analysis and research in this way can obtain different interpretations.
An endogenous phenoloxidase (EPO) from earthworm, Lumbricus rubellus, has been purified and characterized. The purified EPO using ammonium sulfate fractionation, Blue-2, Phenyl-, and Q-sepharose chromatography steps was revealed in SDS-PAGE as a single protein banri with Mr. of 59 kl)a. A native strudure of the enzyme was examined with an in situ staining of a nondenatudng-PAGE using DL-dopa as a substrate. The result showed that a single band due to the EPO activity was located siighdy above a standard polypeptide with Mr. of 210 kl)a. These fads indicate that the EPO is an oligomeric enzyme. The presence of a monophenolase activity of the purified EPO, which hydroxylates tyrosine to dopa, was confirmed by observing dopachrome accumulation at 475 nm at PH 8.0 with a typical lag phase during 60 mm. of meausrement. A series of inhibition study has been performed for the enzyme with several divalent cation chelators such as phenyithiourea (Flu), 1, lO-phenanthroline, EDTA, and EGTA. Among them, only V'flj inhibited the enzyme with 1C0.5 of 65 MM, which indicated that copper was critical for the catalysis of EPO. The enzyme was maximally active at 35'C and pH 8.0 when L-dopa to dopachrome conversion was spectrophotometricaily monitored at 475 nm. The apparent Km values of P0 for L-opa were obtained as 1.86 mM and 13.8 mM at pH 6.5 and 8.0, respectively. The catalytic efficiencies at both pH were almost identical [(kat/Km)pH8.0/(kcat/Km)pH6.5 = O.92] while the Vmax at p11 8.0 was 6.6-fold higher than that at pH 6.5. This fact may indicate that pH affeds the catalysis at substrate and/or enzyme-substrate complex level rather than the enzyme itself. Taken together, the EPO was an oligomeric enzyme which did not require proteolysis for its activation. These results also indicated that the enzyme can exist, at least, in part as a latent form In vivo, which might be distinct from the prophenoloxidase activating system. Therefore, it is pertinent to consider that there must be certain regulatory molecules or phenomena in L. rubellus which make the 1,0 in a latent form in vivo before the foreign invasions.
Journal of the Korean Academy of Child and Adolescent Psychiatry
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v.14
no.1
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pp.12-25
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2003
Objectives:As increasing number of new antidepressants have been being introduced in clinical practice, pharmacological understanding has been broadened. These changes mandate new information and theories to be incorporated into the treatment process of children with depressive disorders. In light of newly coming knowledge, this review intended to recapitulate the characteristics of new antidepressants and to consider the pivotal issues to develope guidelines for the treatment of depression in childhood and adolescence. Methods:Searching the Pub-Med online database for the articles with the key words of 'new', 'antidepressants' and 'children' ninety-seven headings of review articles were obtained. The author selected the articles of pertinent subjects in terms of either treatment guideline or psychopharmacology of new antidepressants. When required, articles about the clinical effectiveness of individual antidepressants were separatedly searched. In addition, the safety information of new antidepressants was acquired by browsing the official sites of the United States Food and Drugs Administration and Department of Health and Human Services. Results:1) For the clinical course, treatment phase, and treatment outcome, the reviews or treatment guidelines adopted the information from adult treatment guidelines. 2) Systematic and critical reviews unambiguously concluded that selective serotonin reuptake inhibitors(SSRIs) excelled tricyclic antidepressants( TCAs) for both efficacy and side effect profiles, and were recommend for the first-line choice for the treatment of children with depressive disorders. 3) New antidepressants generally lacked treatment experiences and randomized controlled clinical trials. 4) SSRIs and other new antidepressants, when used together, might result in pharmacokinetic and/or pharmacodynamic drug-to-drug interaction. 5) The difference of the clinical effectiveness of antidepressants between children and adults should be addressed from developmental aspects, which required further evidence. Conclusion:Treatment guidelines for the pharmacological treatment of childhood and adolescence depression could be constructed on the basis of clinical trial findings and practical experiences. Treatment guidelines are to best serve as the frame of reference for a clinician to make reasonable decisions for a particular therapeutic situation. In order to fulfill this role, guidelines should be updated as soon as new research data become available.
Journal of the korean academy of Pediatric Dentistry
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v.31
no.4
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pp.587-597
/
2004
Over the past 20 years, great strides have been made in research regarding the mechanisms involved in the progression of carious lesions, but new equipment and research tools need to be developed to continue these advancements in caries research. Various methods have been applied to reduce the incidence of carious lesions, which have led to a significant decrease in the number of occlusal caries, but a concurrent increase in the proportion of proximal carious lesions. New diagnostic equipment has been developed to detect early stage carious lesions, and these have demonstrated excellent laboratory results and show promise in clinical applications. The research presented here examines the efficacy of the newly developed $DIFOTI^{TM}$ system in detecting proximal carious lesions compared to traditional intraoral exam and bitewing radiography, possible problems or deficiencies of using the system in clinic, possible improvements that can be made to the system, and the efficacy of detecting early, reversible carious lesions that can be remineralized by preventative fluoride applications. The subject pool consisted of 23 grammer school age patients just prior to entering the mixed dentition phase. Each patient was given a thorough oral examination, radiographic examination consisting of bitewing radiographs of the posterior teeth, and $DIFOTI^{TM}$ examination of the anterior and posterior teeth. Each examination was carried out two times by two examiners, and the data were statistically analyzed. The results are as follows: 1. The mean alpha value of reliability test of the visual oral examination was as follows; occlusal surface was 0.8470. mesial surface was 0.6430, distal surface was 0.5727. lingual surface was 0.2807 and distal surface was 0.2339. When the examination was limited to posterior teeth, the mean alpha value was as follows; occlusal surface was 0.8577, distal surface was 0.8211, lingual surface was 0.7728, buccal surface was 0.7152 and mesial surface was 0.6782. 2. The alpha value of reliability test of the radiographic analysis of carious lesions of the occlusal, mesial, and distal surfaces was 0.8500. 3. The alpha value of reliability test of the $DIFOTI^{TM}$ diagnostic analysis of carious lesions of the occlusal, buccal, lingual, mesial, and distal surfaces was determined to be 0.7917. 4. The $DIFOTI^{TM}$ diagnostic system was found to be the most accurate means of detecting occlusal, buccal, and lingual surface carious lesions (p<0.05), while mesial and distal proximal carious lesions were most accurately assessed using bitewing radiography (p<0.05).
Purpose: There has been a renewal of interest in Macrophage migration inhibitory factor (MIF), especially correlation in pathogenesis of sepsis by many infectious diseases and in regulation of host inflammatory and immune response. We developed immunoradiometric assay (IRMA) to determine serum human MIF concentration. Materials and Methods: The IRMA system utilizes solid phase bound monoclonal anti-recombinant human MIF (rhMIF) antibody as a capture antibody, biotinylated polyclonal anti-rhMIF antibody as a detector antibody. We applied with rhMIF that concentration of standard solutions increased from 0 ng/ml to 100 ng/ml. We used $^{125}I$-streptavidin (SA) as radiotracer to determination of rhMIF concentration. Streptavidin was labeled with $^{125}I$ by Chloramine-T method and $^{125}I$-SA was purified by ultracentrifugation. $^{125}I$-SA stability was evaluated by ITLC analysis at $4^{\circ}C$ and room temperatures until 60days. To validate IRMA system for MIF, we experimented intra-assay and inter-assay coefficients of variation, recovery test and dilution test. Results: Radiolabeling yield of $^{125}I$-SA was 87% and purified $^{125}I$-SA retained above 99% radiochemical purity. $^{125}I$-SA showed above 93% stability in $4^{\circ}C$ until 60days that it is good for immunoradiometric assay as radiotracer. Plotted standard dose response curve showed that increased concentration of rhMIF linearly correlated (R2=0.99) with bound radioactivity of $^{125}I$-SA. The highest intra- and inter-assay coefficients of variation were 5.5% and 7.6%, respectively. The average of recovery of MIF in samples was 102%. In dilution test, linear response curves were obtained (R2=0.97). Conclusion: Radioimmunoassay using $^{125}I$-SA as radiotracer thought to be useful for the determination of serum MIF concentration, and further, its data will be used to evaluate the correlation between clinical significance and serum MIF concentration in patients with various inflammatory diseases.
Ahn Sun Hee;Lee Eun Mi;Kim Dong Gyun;Hong Gyoung Eun;Park Eun Mi;Kong In Soo
Journal of Life Science
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v.16
no.1
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pp.131-135
/
2006
Oligopeptide is known to be an essential nitrogen nutrient for bacterial growth. Oligopeptide can be transported into cytoplasm by a specific transport system, Opp system. Opp system is composed of five proteins, which are transcribed by an operon. These are responsible for oligopeptide binding protein (OppA), permease (OppB and OppC) and energy generation system (OppD and OppF), respectively. Previously, we isolated the opp operon from Vibrio fluvialis and constructed the oppA mutant by allelic exchange method. In this study, we investigated the growth pattern and biofilm production under the different growth condition. When the cells were cultivated using brain heart infusion(BHI) medium, the wild type was faster than the mutant in growth during the exponential phase. However, it showed that the growth pattern of two strains in M9 medium is very similar. The growth of wild type showed better than that of the mutant grown at pH 8. At pH 7, there was no an obvious difference in growth. After 5 mM $H_2O_2$ was treated to the cells $(OD_{600}=1.2)$, the cell survival was examined. The oppA mutation did not affect in survivability. In the presence of $10{\mu}g/ml$ polymyxin B, the biofilm production of the oppA mutant was higher than that of the wild type.
Kim, Sung-Min;Kwon, Jeong-Wook;Ahn, Ki-Chang;Cho, Il-Kyu;Kyung, Kee-Sung;Lee, Jae-Koo
The Korean Journal of Pesticide Science
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v.7
no.3
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pp.176-181
/
2003
The leaching behaviour of [aniline-$^{14}C$]mefenacet in soil was investigated using glass columns (5 cm I.D. $\times$ 30 cm. H) packed with two types of soils with different physicochemical properties. $^{14}C$-Mefenacet (8.33 kBq) and mefenacet (in total, 1.05 mg/kg) were treated onto soil columns and rice plants (Oryza sativa L.) were grown for 17 weeks on these columns. Leachates from the columns were collected at the rate of 122.5 mL per week. $^{14}C$-Activities leached from soil A (OM, 3.1%; CEC, 86 mmol(+)/kg; texture, loam) columns with and without rice plants were 1.95 and 4.19% of the originally applied, whereas those from soil B (OM, 1.3%; CEC, 71 mmol(+)/kg; texture, loam) were 2.69 and 7.05%, respectively. These results indicated that larger amounts of $^{14}C$ were percolated from soil B with less organic matter and from the columns without vegetation. $^{14}C$-Activities absorbed by rice plants from soil A and B were 8.95 and 8.47%, respectively, most of which remained in the root and shoot excluding unhulled grains and ears without grains. $62\sim73%$ of the originally applied $^{14}C$ remained in the depth of $0\sim5cm$ in soil. The mass balance indicated that the losses by volatilization and/or mineralization amounted to $3.4\sim9.2%$ of the originally applied. $^{14}C$-Radioactivities in the aqueous phase of the leachates ranged from 59.4 to 97.7% of the radioactivities in leachates, showing the fast transformation of mefenacet to the polar metabolites.
Purpose : To evaluate changes in expression of cell cycle related genes during apoptosis induced in HL60 cells by X-irradiation to understand molecular biologic aspects in mechanism of radiation therapy. Material and Methods : HL-60 cell line (promyelocytic leukemia cell line) was grown in culture media and irradiated with 8 Gr by linear accelerator (6 MV X-ray). At various times after irradiation, ranging from 3 to 48 hours were analyzed apoptotic DNA fragmentation assay for apoptosis and by western blot analysis and semi-quantitative RT-PCR for expression of cell cycle related genes (cyclin A, cyclin B, cyclin C, cyclin Dl, cyclin E, cdc2, CDK2, CDK4, $p16^{INK4a}$, $p21^{WAF1}$, $p27^{KIP1}$, E2F, PCNA and Rb). Results : X-irradiation (8 Gy) induced apoptosis in HL-60 cell line. Cycline A protein increased after reaching its peak 48 h after radiation delivery and cyclin E, E2F, CDK2 and RB protein increased then decreased after radiation. Radiation induced up-regulation of the expression of E2F is due to mostly increase of Phosphorylated retinoblastoma proteins (ppRb). Cyclin Dl, PCNA, CDC2, CDK4 and $p16^{INK4a}$ protein underwent no significant change at any times after irradiation. There was not detected $p21^{WAF1}$ and $p27^{KIP1}$ protein. Cyclin A, B, C mRNA decreased immediately after radiation and then increased at 12 h after radiation. Cyclin Dl mRNA increased immediately and then decreased at 48 h after radiation. After radiation, cyclin E mRNA decreased with the lapse of time. CDK2 mRNA decreased at 3h and increased at eh after radiation. CDK4 mRNA rapidly increased at 6 to 12 h after radiation. There was no change of expression of $p16^{INK4a}$ and not detected in expressin of $p21^{WAF1}$ and $p27^{KIP1}$ mRNA. Conclusion : We suggest that entry into S phase may contribute to apoptosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of pRb protein are related with radiation induced auoptosis of HL60 cells and tosis of HL60 cells induced by irradiation. Increase of ppRb and decrease of PRb protein are related with radiation induced apoptosis of HL60 cells and this may be associated with induction of E2F and cyclinE/CDK2. These results support that $p21^{WAF1}$ and $p27^{KIP1}$ are not related with radiation induced-apoptosis.
During January 1998-October 1999, the impact of free-floating plants (FFP) on limnology of the wetland ecosystem was evaluated through the investigation of physicochemical characteristics of the Woopo Wetland along with in situ manipulation experiments of aquatic plants. Flooding events occurred in the wetland during the summer period (Jun.-Aug.) and water levels rose to 2-3 m due to precipitation from the catchment and inflow from the main channel of the Nakdong River. Physicochemical parameters and plankton dynamics in the wetland during the summer were greatly influenced by floods and growth of free-floating plants. Dissolved oxygen (Jun.-Sept., 4.5${\pm}$2.5 mg/1; Oct.-May, 8,1 ${\pm}$4.0 mg/1) and pH (Jun.-Sept., 6.9${\pm}$0.4; Oct.-May,7.4${\pm}$0.8) levels were significantly lower during the summer than any other seasons. Three types of enclosure experiments (100 L, treatments with floating plants, screened and opened) were conducted under the presence and absence of sediment for 15 days in the 1999 summer. The treatments with sediment had higher levels of nutrient concentrations than those of the others. Among the treatments with sediment, nutrient concentrations in the treatments with free-floating plants were higher than the others. Zooplankton communities in each treatment showed a similar variation, although the scale of zooplankton densities differed. Rotifer community dominated the zooplankton at the initial phase of the experiment, but decreased drastically along with an increase of cladoceran and copepod communities. In conclusion, low levels of dissolved oxygen and pH in the Woopo Wetland during the summer seemed to be caused by a proliferation of free-floating plants and active decomposition process at the bottom of the sediment.
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