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Establishment of Mouse Embryonic Stem Cell-like Cells from In Vitro Fertilized Embryos (체외수정 생쥐 배아에서의 배아 줄기세포 확립)

  • Shin, Yong-Moon;Park, Yong-Bin;Kim, Hee-Sun;Oh, Sun-Kyung;Chun, Dae-Woo;Suh, Chang-Suk;Choe, Young-Min;Kim, Jung-Gu;Lee, Jin-Yong;Kim, Seok-Hyun
    • Clinical and Experimental Reproductive Medicine
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    • v.29 no.1
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    • pp.1-12
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    • 2002
  • Objective: In order to acquire the technique for the establishment of human embryonic stem cells (ESe) derived from the human frozen-thawed embryos produced in IVF-ET program, this study was performed to establish mouse ESC derived from the in vitro fertilized embryos. Materials and Methods: After Fl hybrid (C57BL female $\times$ CBA mael) female mice were superovulated with PMSG and hCG treatment, their oocytes were retrieved and inseminated, and the fertilized embryos were cultured for 96-120 hours until the expected stages of blastocysts were obtained. To isolate the inner cell mass (ICM), either the blastocysts were treated with immunosurgery, or the whole embryos were cultured for 4 days. Isolated ICMs were then cultured onto STO feeder cell layer, and the resultant ICM colonies were subcultured with trypsin-EDTA treatment. During the subculture process, ESC-like cell colonies were observed with phase contrast microscopy. To identify ESC in the subcultured ESC-like cell colonies, alkaline phosphatase activity and Oct-4 (octamer-binding transcription factor-4) expression were examined by immunohistochemistry and RT-PCR, respectively. To examine the spontaneous differentiation, ESC-like cell colonies were cultured without STO feeder cell layer and leukemia inhibitory factor (LIF). Results: Seven ESC-like cell lines were established from ICMs isolated from the in vitro fertilized embryos. According to the developmental stage, the growth of ICMs isolated from the expanded blastocysts was significantly better than that of ICMs isolated from the hatched blastocysts (80.3% vs. 58.7%, p<0.05). ESC-like cell colonies were only obtained from ICMs of expanded blastocysts. However, the ICMs isolated from the embryos treated with immunosurgery were poorly grown and frequently differentiated during the culture process. The established ESC-like cell colonies were positively stained with alkaline phosphatase and expressed Oct-4, and their morphology resembled that observed in the previously reported mouse ESC. In addition, following the extended in vitro culture process, they maintained their expression of cell surface markers characteristic of the pluripotent stem cells such as alkaline phosphatase and Oct-4. When cultured without STO feeder cell layer and LIF, they were spontaneously differentiated into the various types of cells. Conclusion: The findings of this study suggest that the establishment of mouse ESC can be successfully derived from the in vitro fertilized embryos. The established ESC-like cells expressed the cell surface markers characteristic of the pluripotent stem cells and spontaneously differentiated into the various types of cells.

Effect of Phenolic Mediators and Humic Acid on the Removal of 1-Indanone Using Manganese Oxide (망간산화물(Birnessite)을 이용한 1- Indanone 제거 시 페놀계 반응매개체와 휴믹산(HA) 영향 평가)

  • Choi, Chan-Kyu;Eom, Won-Suk;Shin, Hyun-Sang
    • Journal of Korean Society of Environmental Engineers
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    • v.34 no.7
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    • pp.445-453
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    • 2012
  • An investigation for removal of 1-indanone (1-ID), which were commonly produced from the biological and/or chemical treatment and natural weathering of the PAHs-contaminated soils, via oxidative transformation mediated by birnessite in the presence of various phenolic mediators is described. This study also examines the potential effect of the natural occurring substance humic acid (HA) on the oxidative transformation. The experiment was carried out in aqueous phase as a batch test (10 mg/L 1-ID, 0.3 mM phenolic mediators, $1.0g/L\;{\delta}-MnO_2$, at pH 5). All of the 11 tested phenoilic mediators belong to the group of natural occurring phenols and are widely used as model constituents of humic substances. From the results of HPLC analysis, it is demonstrated that 1-ID was not reactive to birnessite itself, but it can be effectively removed in birnessite-mediated cross coupling reactions in the presence of the phenolic mediators. The percent removals of 1-ID after 2 day incubation were ranged from 9.2 to 71.2% depending on the phenolic mediators applied. The initial rate constant ($K_{int}$, $hr^{-1}$) values for the 1-ID removals obtained from the pseudo-first-order kinetic plots also widely ranged from 0.18 to 15.0. Results of the correlative analysis between the removal efficiencies and structural characteristics of phenolic mediators indicate that the transformation of the 1-ID was considerably enhanced by the addition of electron-donating substituents (e.g., -OH, $-OCH_3$) at the benzne ring, and much less enhanced by the addition of electron-withdrawing substituents (e.g., -COOH, -CHO). The presence of HA showed that removal efficiencies of 1-ID in the birnessite-phenolic mediator systems decreased with increasing HA concentrations. However at low concentration of HA (< 2 mg/L), it caused some enhancement in the removals of 1-ID as compared to the control.

Choledochal Cyst in Korea - A Survey by the Korean Association of Pediatric Surgeons - (담관낭종 -대한소아외과학회회원 대상 전국조사-)

  • Choi, Kum-Ja;Kim, D.Y.;Kim, S.Y.;Kim, S.C.;Kim, S.K.;Kim, W.K.;Kim, I.K.;Kim, J.E.;Kim, J.C.;Kim, H.Y.;Kim, H.J.;Park, K.W.;Park, W.H.;Park, J.Y.;Paek, H.K.;Seo, J.M.;Song, Y.T.;Oh, S.M.;Yoo, S.Y.;Lee, D.S.;Lee, M.D.;Lee, S.K.;Lee, S.C.;Park, Y.S.;Lee, T.H.;Chung, S.Y.
    • Advances in pediatric surgery
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    • v.9 no.1
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    • pp.45-51
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    • 2003
  • A nationwide survey on choledochal cyst was undertaken among 39 members of the Korean Association of Pediatric Surgeons. The members were required to complete a questionnaire and the case registration form for each patient during the five year period of 1997 to 2001. Three hundred and forty eight patients were registered from 32 institutions. The average number of patients per surgeon was one to two every year. The male to female ratio was 1:3.4. The age of patients on diagnosis was $49.0{\pm}44.4$ months. The geographic distribution was 34.8% in Seoul and Kyoungki-do, 33.3% in Kyoungsang-do, 17.9% in Cholla-do, and 8.5% in Choongchung-do, in order of frequency. The three common clinical presentations were abdominal pain (63.8%), vomiting (35.3%), and jaundice (29.1%). Only seven patients (2%) presented with classic triad, and 25 patients were diagnosed by antenatal ultrasonographic examination. According to the Todani Classification, 238 patients (7l.3%) were type 1, 3 (0.9%) type 11, and 93 (27.8%) type IV. At the time of the operation, three important associated conditions were choledocholithiasis in 45 patients (15.1%), liver fibrosis (Grade 1-4) in 35, and previous operative procedure for biliary diseases in 10. Associated anomalies were observed in 13 patients (3.8%). Three hundred thirty nine (98.8%) of 343 lesions were treated by Cyst excision and Roux-Y hepaticoiejunostomy. One hundred seventy-six patients had an anomalous arrangement of the pancreatobiliary ductal system (APBD): APBD was not in 92 patients, biliary duct joined to the pancreatic duct in 51, and pancreatic duct joined to the biliary duct in 26. There were 8.5% early, and 7.7% late phase operative complications. The major complications were bleeding, anastomotic leakage, and acute pancreatitis. The combination of acute abdomen and choledochal cyst may suggest spontaneous rupture. Because of the development of late intrahepatic bile duct stones, long term follow up after cyst excision and hepaticojejunostomy is required. The optimal time of surgical intervention should also be considered in the situation of routine use of antenatal ultrasonographic examination. This is the first review of the choledochal cyst in Korea and provides baseline data for future comparisons.

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Perfluoropolymer Membranes of Tetrafluoroethylene and 2,2,4Trifluofo- 5Trifluorometoxy- 1,3Dioxole.

  • Arcella, V.;Colaianna, P.;Brinati, G.;Gordano, A.;Clarizia, G.;Tocci, E.;Drioli, E.
    • Proceedings of the Membrane Society of Korea Conference
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    • 1999.07a
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    • pp.39-42
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    • 1999
  • Perfluoropolymers represent the ultimate resistance to hostile chemical environments and high service temperature, attributed to the presence of fluorine in the polymer backbone, i.e. to the high bond energy of C-F and C-C bonds of fluorocarbons. Copolymers of Tetrafluoroethylene (TEE) and 2, 2, 4Trifluoro-5Trifluorometoxy- 1, 3Dioxole (TTD), commercially known as HYFLON AD, are amorphous perfluoropolymers with glass transition temperature (Tg)higher than room temperature, showing a thermal decomposition temperature exceeding 40$0^{\circ}C$. These polymer systems are highly soluble in fluorinated solvents, with low solution viscosities. This property allows the preparation of self-supported and composite membranes with desired membrane thickness. Symmetric and asymmetric perfluoropolymer membranes, made with HYFLON AD, have been prepared and evaluated. Porous and not porous symmetric membranes have been obtained by solvent evaporation with various processing conditions. Asymmetric membranes have been prepared by th wet phase inversion method. Measure of contact angle to distilled water have been carried out. Figure 1 compares experimental results with those of other commercial membranes. Contact angles of about 120$^{\circ}$for our amorphous perfluoropolymer membranes demonstrate that they posses a high hydrophobic character. Measure of contact angles to hexandecane have been also carried out to evaluate the organophobic character. Rsults are reported in Figure 2. The observed strong organophobicity leads to excellent fouling resistance and inertness. Porous membranes with pore size between 30 and 80 nanometers have shown no permeation to water at pressures as high as 10 bars. However high permeation to gases, such as O2, N2 and CO2, and no selectivities were observed. Considering the porous structure of the membrane, this behavior was expected. In consideration of the above properties, possible useful uses in th field of gas- liquid separations are envisaged for these membranes. A particularly promising application is in the field of membrane contactors, equipments in which membranes are used to improve mass transfer coefficients in respect to traditional extraction and absorption processes. Gas permeation properties have been evaluated for asymmetric membranes and composite symmetric ones. Experimental permselectivity values, obtained at different pressure differences, to various single gases are reported in Tab. 1, 2 and 3. Experimental data have been compared with literature data obtained with membranes made with different amorphous perfluoropolymer systems, such as copolymers of Perfluoro2, 2dimethyl dioxole (PDD) and Tetrafluorethylene, commercialized by the Du Pont Company with the trade name of Teflon AF. An interesting linear relationship between permeability and the glass transition temperature of the polymer constituting the membrane has been observed. Results are descussed in terms of polymer chain structure, which affects the presence of voids at molecular scale and their size distribution. Molecular Dyanmics studies are in progress in order to support the understanding of these results. A modified Theodoru- Suter method provided by the Amorphous Cell module of InsightII/Discover was used to determine the chain packing. A completely amorphous polymer box of about 3.5 nm was considered. Last but not least the use of amorphous perfluoropolymer membranes appears to be ideal when separation processes have to be performed in hostile environments, i.e. high temperatures and aggressive non-aqueous media, such as chemicals and solvents. In these cases Hyflon AD membranes can exploit the outstanding resistance of perfluoropolymers.

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Sources Identification of Anthropogenic Pb in Ulleung Basin Sediments using Stable Pb Isotope Ratios, East/Japan Sea (동해 울릉분지 시추 퇴적물에서 안정 Pb 동위원소를 이용한 Pb의 기원 추정)

  • Choi, Man-Sik;Uoo, Jun-Sik;Kim, Dong-Seon
    • The Sea:JOURNAL OF THE KOREAN SOCIETY OF OCEANOGRAPHY
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    • v.12 no.4
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    • pp.315-327
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    • 2007
  • This study investigated temporal and spatial variation of Pb and stable Pb isotopes accumulated in Ulleung Basin core sediments (4) using MC ICP/MS in order to identify the sources of anthropogenic Pb in the East/Japan Sea. Leached (1M HCl) Pb concentration and isotope ratios ($^{207}Pb/^{206}Pb\;and\;^{208}Pb/^{206}Pb$) were nearly constant during 300 yrs past than 1930, but increased up to twice in concentration and as much as 3.41% (1.70%) after 2000. On the other hand, residual Pb concentrations were nearly constant for past 400 yrs. The accumulation rates of anthropogenic Pb in the basin area were in the range of $3.1-3.5mg/m^2/yr$, which were similar levels to total atmospheric Pb deposition fluxes from 1990s to the present. In the slope area, more increase of anthropogenic Pb accumulation than the levels expected from mass accumulation rate could be found after the middle of 1990s. From the detailed evaluation for the temporal and spatial variation of accumulation rate and isotope ratios of anthropogenic Pb, we proposed probable sources and pathways of anthropogenic Pb. Pb emmision by coal burning from the China and Korea initiated the accumulation of anthropogenic Pb in the sediments of East/Japan Sea from 1930s. The accumulation of Pb increased by the addition of anti-nocking agents from both countries untill the beginning of 1990s, but from the middle of 1990s to the present, the phase-out of gasoline additives and the rapid increase of coal burning from the China maintained the atmospheric Pb levels in the Ulleung basin nearly similar to before. However, the local sources within this basin might take an important role in the rapid increase of anthropogenic Pb accumulation in slope areas from the middle of 1990s.

Sorption Characteristics of Cs on Weathered Biotite (흑운모 풍화에 따른 Cs 이온의 흡착 특성)

  • Kim, Ji-Yeon;Kim, Yeongkyoo
    • Journal of the Mineralogical Society of Korea
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    • v.28 no.3
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    • pp.255-263
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    • 2015
  • To investigate the effect of the weathering process of biotite on Cs sorption, sorption experiment of Cs with $10^{-3}M$ solution was carried out on the biotite reacted at different reaction times at pH 2 and 4, and 1 M solutions of Na, K, Ca, Mg, Rb, and Cs. Peak changes were observed for some samples by XRD, indicating that new mineral phase formed by biotite weathering. Among several factors, cations in solutions have the most significant influence on the mineralogical changes. The samples reacted with Na showed the most outstanding change with increasing peak width and appearance of $12{\AA}$ peak and $14{\AA}$ peak. This new peaks indicate the formation of hydrobiotite and vermiculite. The new peaks had stronger peak intensity for the sample reacted at pH 4 than that at pH 2, probably due to the fast dissolution of small particles and edges and resultant decrease in the formation of expandable layers. The biotite reacted at Mg solution showed small intensity at $14{\AA}$. Based on XRD results, the degree of biotite weathering was in the order of Na, Mg, and Ca. The samples reacted with K, Rb, Cs solutions did not show noticeable mineralogical changes caused by weathering. The amount of sorbed Cs on weathered biotite showed close relationship with the degree of weathering indicated by XRD. At both pH 2 and 4, the biotite reacted with Na solution showed the highest Cs sorption, and those with Mg and Ca solutions showed the next highest ones. The sorbed amounts of Cs on the bitote reacted with K, Rb, Cs solutions were relatively low. This indicates that at the Cs concentration ($10^{-3}M$) which we used for this experiment and which was much higher than the maximum Cs concentration sorbed on the frayed edge site, expandable layer plays more important role than frayed edge. In the cases of K, Rb, and Cs solutions, Cs sorption was decreased because K is the same cations as the one in the interlayer or the sorption of Rb and Cs on the frayed edge prevents the formation of expandable layers.

Behaviour of the Soil Residues of the Acaricide-Insecticide, [$^{14}C$]Acrinathrin;I. Behaviour during Crop(Maize) Cultivation (살비살충제 [$^{14}C$Acrinathrin 토양 잔류물의 행적 규명;I. 농작물(옥수수) 재배시의 행적)

  • Lee, Jae-Koo;Kyung, Kee-Sung;Kwon, Jeong-Wook;Ahn, Ki-Chang;Jung, In-Sang
    • Korean Journal of Environmental Agriculture
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    • v.14 no.2
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    • pp.186-201
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    • 1995
  • In order to elucidate the fate of the residues of the pyrethroid acaricide-insecticide, acrinathrin in soil, maize plants were grown for one month on the specially-made pots filled with two different types of soils containing fresh and one-month-aged residues of [$^{14}C$]acrinathrin, respectively. The mineralization of [$^{14}C$]acrinathrin to $^{14}CO_2$ during the one-month period of aging and of maize cultivation amounted to $23{\sim}24%$ and $24{\sim}33%$, respectively, of the original $^{14}C$ activities. At harvest after one-month growing, the shoots and roots contained less than 0.1% and 1% of the originally applied $^{14}C$ activity, respectively, whereas the $^{14}C$ activity remaining in soil was $65{\sim}80%$ in both soils. Three degradation products with m/z 198(3-phenoxybenzaldehyde), m/z 214(3-phenoxybenzoic acid), and m/z 228(methyl 3-phenoxybenzoate) besides an unknown were identified from acetone extracts of both soils without and with maize plants after treatment of [$^{14}C$]acrinathrin, by autoradiography and GC-MS, and those with m/z 225(3-phenoxybenzaldehyde cyanohydrin) and m/z 198 (3-phenoxybenzaldehyde) from acetone extract of the Soil A treated with 50 ppm acrinathrin and grown with maize plants for 30 days were identified by mass spectrometry. These results suggested that the hydrolytic cleavage of the ester linkage adjacent to the $^{14}C$ with a cyano group, forming 3-phenoxybenzaldehyde cyanohydrin. The removal of hydrogen cyanide therefrom leads to the formation of 3-phenoxybenzaldehyde as one of the major products. The subsequent oxidation of the aldehyde to 3-phenoxybenzoic acid, followed by decarboxylation would evolve $^{14}CO_2$. Solvent extractability of the soils where maize plants were grown for 1 month and/or [$^{14}C$]acrinathrin was aged for 1 month was less than 31% of the original $^{14}C$ activity and over 95% of the total $^{14}C$ activity in soil extracts was distributed in the organic phase. Accordingly, acrinathrin turned out to be degraded rapidly in both soils and be bound to soil constituents as well, not being available to crops.

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Effects of Initial Shoot, Root Length, and Acclimating Substrates on Survival Rate of Plantlets Regenerated from Somatic Embryos of Larix kaempferi (일본잎갈나무 체세포배 유래 식물체의 초기 신초와 뿌리 길이, 순화용 기질이 생존율에 미치는 영향)

  • Lee, Na Nyum;Yun, A Young;Kim, Ji Ah;Kim, Tae Dong;Kim, Yong Wook;Han, Sim Hee
    • Journal of Korean Society of Forest Science
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    • v.109 no.4
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    • pp.413-420
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    • 2020
  • We analyzed the growth characteristics of each cell line and acclimating substrate of Larix kaempferi plantlets regenerated from somatic embryos, with the goal of increasing the survival rate during the acclimation phase. Somatic embryos from three embryogenic cell lines (L14-66, L16-18, and L17-B4) were used, and the acclimating substrates were commercial soils for Larix species (Larix-Soil) and horticultural corps (Hort-Soil), Elle-pot, and Peat-plug. The average initial shoot and root length was shortest in L14-66 and longest in L17-B4. The average survival rate by cell line was highest (87.0%) in L17-B4 and lowest (64.3%) for L14-66. Survival rates by substrate were highest in Elle-pot (88.5%) and Peat-plug (88.9%). The initial shoot length of the L14-66 plantlets was highly correlated with survival rates in the Larix-Soil (r = 0.852), Hort-Soil (r = 0.692), and Elle-pot (r = 0.867) substrates, but not in Peat-plug with high total nitrogen content. The initial shoot length of the L17-B4 plantlets was not correlated with the survival rate in any of the substrates. The initial root length of the L14-66 plantlets was highly related to survival rates in the Larix-Soil (r = 0.986), Elle-pot (r = 0.846), and Peat-plug (r = 0.802) substrates, and the survival rate of the plantlets was higher as the initial root length was longer. The initial root length of the L17-B4 plantlets was related to survival rate only in the Larix-Soil (r = 0.896) and Elle-pot (r = 0.696) substrates. In conclusion, to increase the survival rate of plantlets, root length should be considered over shoot length, and it is recommended to use substrates with high nitrogen content, such as Peat-plug, or to add fertilizer, during the acclimating process. In addition, in order to increase the survival rate, lines with high initial growth should be developed.

Benthic Foraminiferal Assemblage and Sedimentary Environment of Core Sediments from the Northern Shelf of the East China Sea (북동중국해 대륙붕 코아 퇴적물의 저서유공충 군집 특성과 퇴적환경 연구)

  • Kang, So-Ra;Lim, Dhong-Il;Kim, So-Young;Rho, Kyoung-Chan;Yoo, Hae-Soo;Jung, Hoi-Soo
    • Journal of the Korean earth science society
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    • v.29 no.6
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    • pp.454-465
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    • 2008
  • Benthic foraminiferal assemblage and AMS radiocarbon dating of core sediments from the northern shelf of the East China Sea were analyzed in order to understand the paleoenvironment and sedimentary environmental changes around the Korean marginal seas since the last glacial maximum (LGM). The core sediments, containing continuous records of the last 16,000 years, reveal a series of well-defined vertical changes in number of species (S), P/T ratio and species diversity (H) as well as foraminiferal assemblage. Such down-core variations display a sharp change at a core depth of approximately 240 cm, which corresponds to ca. 10,000 year B.P. The sediments of the lower part of the core (240${\sim}$560 cm, Zone I), including the well-developed tide-influenced sedimentary structures, are characterized by high abundances of Ammonia beccarii and Elphidium clavatum (s.l.) and low values in number of species, P/T ratio and diversity. These tide-influenced signatures and foraminiferal assemblage characters suggest that the sediments of Zone I were deposited in a coastal environment (water depths of 20${\sim}$30 m) such as tidal estuary with an influence of the paleo-rivers (e.g., old-Huanghe and Yangtze rivers) during the early phase of the sea-level rise (ca. 16,000 to 10,000 years) since the LGM. In contrast, the upper core sediments (0${\sim}$240 cm, Zone II) are characterized by abundant Eilohedra nipponica and Bolivina robusta with a minor contribution of A. ketienziensis angulata and B. marginata. and high values in number of species, P/T ratio and diversity. Based on relative abundance of these assemblage, Zone II can be divided into two subzones (IIa and IIb). Zone IIa is interpreted to be deposited under the inner-to-middle shelf environment during the marine transgression in the early Holocene (after ca. 9,000 yr B.P.) when sea level rapidly increased. The sediments of zone IIb most likely deposited after 6,000 yr B.P. under the outer shelf environment (80${\sim}$100 m water depth), which is similar to modem depositional environments. The muddy sediments of zone IIb were probably transported from the old-Huanghe and Yangtze Rivers during the late Holocene. We suggest that the present-day oceanographic conditions over the Yellow and the East China Seas have been established after ca. 7,000${\sim}$6,000 yr B.P. when the Kuroshio Current began to influence this area.

Dynamics of Barrel-Shaped Young Supernova Remnants (항아리 형태 젊은 초신성 잔해의 동력학)

  • Choe, Seung-Urn;Jung, Hyun-Chul
    • Journal of the Korean earth science society
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    • v.23 no.4
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    • pp.357-368
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    • 2002
  • In this study we have tried to explain the barrel-shaped morphology for young supernova remnants considering the dynamical effects of the ejecta. We consider the magnetic field amplification resulting from the Rayleigh-Taylor instability near the contact discontinuity. We can generate the synthetic radio image assuming the cosmic-ray pressure and calculate the azimuthal intensity ratio (A) to enable a quantitative comparison with observations. The postshock magnetic field are amplified by shearing, stretching, and compressing at the R-T finger boundary. The evolution of the instability strongly depends on the deceleration of the ejecta and the evolutionary stage of the remnant. the strength of the magnetic field increases in the initial phase and decreases after the reverse shock passes the constant density region of the ejecta. However, some memory of the earlier phases of amplification is retained in the interior even when the outer regions turn into a blast wave. The ratio of the averaged magnetic field strength at the equator to the one at the pole in the turbulent region can amount to 7.5 at the peak. The magnetic field amplification can make the large azimuthal intensity ratio (A=15). The magnitude of the amplification is sensitive to numerical resolution. This mens the magnetic field amplification can explain the barrel-shaped morphology of young supernova remnant without the dependence of the efficiency of the cosmic-ray acceleration on the magnetic field configuration. In order for this mechanism to be effective, the surrounding magnetic field must be well-ordered. The small number of barrel-shaped remnants may indicate that this condition rarely occurs.