• Title/Summary/Keyword: Co-incubated

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Effects of Increased CO2 and Temperature on the Growth of Four Diatom Species (Chaetoceros debilis, Chaetoceros didymus, Skeletonema costatum and Thalassiosira nordenskioeldii) in Laboratory Experiments

  • Hyun, Bonggil;Choi, Keun-Hyung;Jang, Pung-Guk;Jang, Min-Chul;Lee, Woo-Jin;Moon, Chang-Ho;Shin, Kyoungsoon
    • Journal of Environmental Science International
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    • v.23 no.6
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    • pp.1003-1012
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    • 2014
  • We examined the combined impacts of future increases of $CO_2$ and temperature on the growth of four marine diatoms (Skeletonema costatum, Chaetoceros debilis, Chaetoceros didymus, Thalassiosira nordenskioeldii). The four strains were incubated under four different conditions: present ($pCO_2$: 400ppm, temperature: $20^{\circ}C$), acidification ($pCO_2$: 1000ppm, temperature: $20^{\circ}C$), global warming ($pCO_2$: 400ppm, temperature: $25^{\circ}C$), and greenhouse ($pCO_2$: 1000ppm, temperature: $25^{\circ}C$) conditions. Under the condition of higher temperatures, growth of S. costatum was suppressed, while C. debilis showed enhanced growth. Both C. didymus and T. nodenskioldii showed similar growth rates under current and elevated temperature. None of the four species appeared affected in their cell growth by elevated $CO_2$ concentrations. Chetoceros spp. showed increase of pH per unit fluorescence under elevated $CO_2$ concentrations, but no difference in pH from that under current conditions was observed for either S. costatum or T. nodenskioeldii, implying that Chetoceros spp. can take up more $CO_2$ per cell than the other two diatoms. Our results of cell growth and pH change per unit fluorescence suggest that both C. debilis and C. didymus are better adapted to future oceanic conditions of rising water temperature and $CO_2$ than are S. costatum and T. nodenskioeldii.

Susceptibility of various Helicobacter pyloris to New Antiulcer Agents

  • Tae W. Woo;Han Y. Yoo;Man S. Chang;Young K. Chung;Kim, Kyu B.;Sang K. Son;Kim, Sung K.;Dae P. Kang;Park, Whan S.
    • Proceedings of the Korean Society of Applied Pharmacology
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    • 1996.04a
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    • pp.197-197
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    • 1996
  • Helicobacter pylori (H. pylori) is a microaerophilic spiral bacterium and infection by it in the human stomach causes gastritis, furthermore, it is considered to be involved in the pathogenesis of peptic ulcers and the development of gastric carcinoma. We assessed the inhibitory activity of new antiulcer drugs against Helicobacter pylori. The activities of new antiulcer agents against Helicobacter pylori strains were determined by the standard agar dilution method with blood agar base #2, supplemented with 5% sheep blood and 4 antibiotics to support growth of these organisms. They were inoculated by multipoint inoculator and incubated at 37$^{\circ}C$ for 3 days under microaerophilic atmosphere. The MIC of antiulcer agents was the lowest concentration that inhibited visible growth of these organisms. According to results of various biochemical tests, these bacteria were identified as Helicobacter pylori strains. And the MIC results showed that the strains were very susceptible to omeprazole and YJA20379s. Some of YJA20379s were more potent than omeprazole. These results suggest that our new antiulcer drugs have potent inhibitory activity against Helicobacter pylori, so that our new antiulcer drugs might be useful for the clinical eradication of gastrointestinal Helicobacter pylori.

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Intestinal Absorption of Fibrinolytic and Proteolytic Lumbrokinase Extracted from Earthworm, Eisenia andrei

  • Yan, Xiang Mei;Kim, Chung-Hyo;Lee, Chul-Kyu;Shin, Jang-Sik;Cho, Il-Hwan;Sohn, Uy-Dong
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.2
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    • pp.71-75
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    • 2010
  • To investigate the intestinal absorption of a fibrinolytic and proteolytic lumbrokinase extracted from Eisenia andrei, we used rat everted gut sacs and an in situ closed-loop recirculation method. We extracted lumbrokinase from Eisenia andrei, and then raised polyclonal antibody against lumbrokinase. Fibrinolytic activity and proteolytic activity in the serosal side of rat everted gut sacs incubated with lumbrokinase showed dose- and time-dependent patterns. Immunological results obtained by western blotting serosal side solution using rat everted gut sacs method showed that lumbrokinase proteins between 33.6 and 54.7 kDa are absorbed mostly by the intestinal epithelium. Furthermore, MALDI- TOF mass spectrometric analysis of plasma fractions obtained by in situ recirculation method confirmed that lumbrokinase F1 is absorbed into blood. These results support the notion that lumbrokinase can be absorbed from mucosal lumen into blood by oral administration.

Effect of Co-Culture Mouse Fetal Fibroblast Cell on In Vitro Development of Blastomeres Separated from Mouse Preimplantation Embryos (생쥐 태아 Fibroblast 세포와 공동배양이 초기 생쥐배 분할구의 체외 발생능에 미치는 영향)

  • 김진호;정병헌;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.16 no.4
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    • pp.341-346
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    • 1993
  • The development of isolated blastomeres from mammalian preimplantation embryos has been basically studied for the multiplication of embryos from superior animals. Therefore, this study was investigated the effect of co-culture with mouse fetal fibroblast cells(MFFC) on in vitro development of blastomeres from mouse preimplantation embryos. Mature female ICR mice were treated with hormone to induce superovulation and embryos were collected at each 2, 4, and 8-cell stage. Then, after removing zona pellucida with protease, blastomeres were isolated by micropipetting, or reconstituted with different stage blastomere, and incubated for 72 hrs either in T6 or TCM199 or on the monolayer of MFFC, which was prepared with fibroblast cells from 14∼14 day mouse fetus. After incubation, we examined their development rates every day and the nuclei numbers of each blastocyst by Hoechst-33342 staining. In the development rates of blastomeres, there were no significant differences between media but the higher rateswere found in the monolayer of MFFC, regardless of reconsititution. In addition, blastomeres cultured with MFFC had slightly greater number of nuclei than those cultured in single media. Generally, the higher development rates of blastomeres were found from earlier stage embryos than the later ones, regardless of culture conditions. Reconsitituted blastomeres had more nuclei but did not show the higher development rates, compared to the single blastomeres. Taken together, our results suggest that co-culture with MFFC have a beneficial effect on the in vitro development of blastomeres from mouse embryos.

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Effect of Photosynthesis on Ozone-Induced Ethylent Evolution from Tomato Plants (토마토 식물에 있어서 광합성이 유존유동성의 에틸렌 생성에 미치는 영향)

  • 배공영
    • Journal of Korean Society for Atmospheric Environment
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    • v.12 no.3
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    • pp.307-314
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    • 1996
  • The rate of evolution of ethylent by tomato plants was rapidly increased by ozone fumigation. In the present study, the mechanism of ethylent evolution by ozone was investigated in experiments with aminoethoxyvinylglycine (AVG) and tiron, which inhibit the formation of ethylene and peroxidation of lipids, respectively. Pretreatment with AVG significantly inhibited the ozone-induced ethylent evolution, but the treatment of plants with tiron did not inhibit. These results indicate that the induction of the evolution of ethylene by ozone involves the pathway via aminocyclopropane-1-carboxylate (ACC), while not released as a result of the peroxidation of lipids. Ozone-induced ethylent evolution was greater in dar- than light-incubated, intact tomato plants. The difference between dark- and light-ethylene evolution was examined with diuron, an inhibitor of photosynthetic electron transport. The inhibitor treatment promoted ethylent evolution. These results suggest that ethylent retention and metabolism in plants were regulated by internal $CO_2$ levels which, in turn, were controlled in large part by photosynthesis. Thus, ethylene was retained in illuminated leaf tissue under low intenal $CO_2$ concentration which may develop in a sealed container without exogenously supplied $CO_2$.

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Particle Size Effects in Buffer System using Calcium Carbonate Bead Immobilized with Alginate for the Cultivation of Bifidobacterium (Alginate에 고정화된 Calcium Carbonate를 이용한 Buffer System에서 비드 크기에 따른 Bifidobacterium의 배양에 대한 효과)

  • Lee, Ki-Yong;Heo, Tae-Ryeon
    • Korean Journal of Food Science and Technology
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    • v.30 no.2
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    • pp.425-433
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    • 1998
  • The characteristics of the reaction of calcium carbonate $(CaCO_3)$ immobilized with alginate as buffer system for the high concentration cultivation of bifidobacteria in fermenter are described by the mathematical model, and tested for the reusing possibility of the used $CaCO_3$ beads. When$CaCO_3$ beads with the various diameters were reacted in 0.1 M of the mixed organic acids (0.6 M of acetic acid and 0.4 M lactic acid) and in fermenter inoculated Bifidobacterium longum ATCC 15707, the change of bead diameters can be calculated with the amount of the decreased $CaCO_3$ from the surface of bead using the mathematical model. These values was similar to the directly measured bead diameter by a micrometer. Therefore, it was considered that the mathematical model could be used for explaining the reaction charateristics of the $CaCO_3$ bead reacted with the organic acids. When Bifidobacterium longum was incubated at $37^{\circ}C$ for 20 hours in fermenter with $CaCO_3$ beads, the buffering effect of $CaCO_3$, the reduce rate of the bead diameter, and the growth rate of Bifidobacterium longum were higher at the smaller beads than beads with the larger diameters. Also, when Bifidobacterium longum was incubated in fermenter with the mixed beads which were added new beads to the recovered beads in order to equalize with the total surface area of initial beads, the buffering effect of $CaCO_3$ bead and the growth rate of Bifidobacterium longum were very corresponded with the results of the fermentation using the only initial beads. Therfore, it is expected that the used beads can be reused by adding the initial beads.

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In vivo and In vitro Metabolism of Recombinant Human Epidermal Growth Factor (DWP401) in Rats (재조합 인간 상피세포성장인자(DWP401)의 흰쥐에서의 in vivo와 in vitro 대사)

  • Koh, Yeo-Wook;Nam, Kouen-Ho;Jung, Ju-Young;Park, Seung-Kook;Yu, Young-Hyo;Kim, Jae-Hwan;Han, Kun;Park, Myung-Hwan;Shim, Chang-Koo
    • YAKHAK HOEJI
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    • v.41 no.3
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    • pp.381-388
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    • 1997
  • Metabolism of DWP401, recombinant juman epidermal growth factor, was examined in vivo and in vitro in rats. When $^{125}I$-labeled DWP401 was administered at a dose of 50 ${\mu}g$/kg by i.v. injection. $^{125}I$-labeled DWP401 was rapidly degraded within 30 minytes above 93%. Thin layer chromatography analysis of urine collected for 24 hr after i.v. administration of $^{125}I$-labeled DWP401 showed ohly one spot on a X-ray film which was considered as diiodo-tyrosine. This result suggests tha $^{125}I$-labeled DWP401 was completely digested into free amino acids without any specific intermediate polypeptides. About 42.1% of the administered iodine was recovered in 24 hr. For in vitro degradation study, $^{125}I$-labeled DWP401 was added to plama and tissue homogenates of rats and incubated at $37^{\circ}C$. Almost 98% of the added radioactivity recovered from the protein fraction of the liver, kidey, small intestine, stomach and spleen decreased rapidly. For examplem the recovery rates of $^{125}I$-labeled DWP401 were 58.6, 63.2, 39.9, 52.9 and 66.8% after 4hrs of incubation in respective organ homogenates.

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Effects of Bovine Cumulus Cell Co-Culture and CR1aa Medium on In Vitro Development of In Vitro Produced Bovine Embryos (우 난구세포의 공동배양과 CR1aa배양액이 체외생산된 우 수정란의 체외 발생에 미치는 영향)

  • 김동훈;정형민;박세필;이훈택;정길생
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.271-278
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    • 1994
  • The aim of this study was to compare the two culture systems 1) co-culture with cumulus cells and 2) chemically defined medium supplemented with amino acids (CR1aa) and fetal calf serum (FCS) of in vitro produced bovine embryos from follicular oocytes in vitro. Bovine follicular oocytes were collected from ovaries of slaughtered cows and matured in TCM199 supplemented with 10% FCS and hormones (1$\mu\textrm{g}$/ml FSH-P and 1$\mu\textrm{g}$/ml oestradiol-17$\beta$)24 hours at 39$^{\circ}C$ under 5% CO2 in air. The capacitation of spermatozoa from ejaculated or frozen bull semen was induced by centrifugation through Percoll density gradient (45%, 90%). Then capacitated spermatozoa (1$\times$106/ml) were inseminated into 50${mu}ell$ droplet containing matured follicular oocytes and incubated for 40~42 hours. Cleaved embryos of 2~4cell stage were transferred to the co-culture with cumulus cells and/or CR1aa medium supplemented with FCS. In semen source, the developmental rates to the blastocyst and the hatched blastocyst stages were higher in ejaculated semen(27.6% and 14.9%) than those of frozen-thawed semen(18.3% and 11.8%), respectively. In two culture systems, the proportions of embryonic development upto the blastocysts and the hatched blastocysts were higher of CR1aa medium (22.1% and 12.1%) than those of cumulus cell co-culture (16.8% and 5.1%), respectively. The number of cells in exapnded blastocysts was slightly higher in cumulus cells co-culture (122.6$\pm$8.5) than that in CR1aa medium (117.9$\pm$5.9). The present results indicated that the early development of in vitro produced bovine embryos can be maintained efficiently in CR1aa medium as well as in co-culture with cumulus cells.

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Fertilizer and Organic Inputs Effects on CO2 and CH4 Emission from a Soil under Changing Water Regimes (토양 수분 변동 조건에서 시비 및 유기물 투입에 따른 CO2와 CH4 방출 특성)

  • Lim, Sang-Sun;Choi, Woo-Jung;Kim, Han-Yong
    • Korean Journal of Environmental Agriculture
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    • v.31 no.2
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    • pp.104-112
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    • 2012
  • BACKGROUND: Agricultural inputs (fertilizer and organic inputs) and water conditions can influence $CH_4$ and $CO_2$ emission from agricultural soils. This study was conducted to investigate the effects of agricultural inputs (fertilizer and organic inputs) under changing water regime on $CH_4$ and $CO_2$ emission from a soil in a laboratory incubation experiment. METHODS AND RESULTS: Four treatments were laid out: control without input and three type of agricultural inputs ($(NH_4)_2SO_4$, AS; pig manure compost, PMC; hairy vetch, HV). Fertilizer and organic inputs were mixed with 25 g of soil at 2.75 mg N/25 g soil (equivalent to 110 kg N/ha) in a bottle with septum, and incubated for 60 days. During the first 30-days incubation, the soil was waterlogged (1 cm of water depth) by adding distilled water weekly, and on 30 days of incubation, excess water was discarded then incubated up to 60 days without addition of water. Based on the redox potential, water regime could be classified into wetting (1 to 30 days), transition (31 to 40 days), and drying periods (41 to 60 days). Across the entire period, $CH_4$ and $CO_2$ flux ranged from 0 to 13.8 mg $CH_4$/m/day and from 0.4~1.9 g $CO_2$/m/day, and both were relatively higher in the early wetting period and the boundary between transition and drying periods. During the entire period, % loss of C relative to the initial was highest in HV (16.4%) followed by AS (8.1%), PMC (7.5%), and control (5.4%), indicating readily decomposability of HV. Accordingly, both $CH_4$ and $CO_2$ fluxes were greatest in HV treatment. Meanwhile, the lower $CH_4$ flux in AS and PMC treatments than the control was ascribed to reduction in $CH_4$ generation due to the presence of oxidized compounds such as ${SO_4}^{2-}$, $Fe^{3+}$, $Mn^{4+}$, and ${NO_3}^-$ that compete with precursors of $CH_4$ for electrons. CONCLUSION: Green manure such as HV can replace synthetic fertilizer in terms of N input, however, it may increase $CH_4$ emission from soils. Therefore, co-application of green manure and livestock manure compost needs to be considered in order to achieve satisfactory N supply and to mitigate $CH_4$ and $CO_2$ emission.

In vitro maturation of ovine oocyte in a modified granulosa cells co-culture system and alpha-tocopherol supplementation: effects on nuclear maturation and cleavage

  • Adeldust, Hamideh;Zeinoaldini, Saeed;Kohram, Hamid;Roudbar, Mahmoud Amiri;Joupari, Morteza Daliri
    • Journal of Animal Science and Technology
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    • v.57 no.8
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    • pp.27.1-27.6
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    • 2015
  • This study was designed to investigate the effects of ${\alpha}$-tocopherol and granulosa cells monolayer on nuclear maturation and cleavage rates of ovine cumulus-oocyte complexes (COCs). The COCs (n = 2814) were matured in maturation medium supplemented with various concentration of ${\alpha}$-tocopherol (0, 5, 10, $15{\mu}g/ml$), oocytes were incubated at $39^{\circ}C$ with 5 % $CO_2$ for 24 h in three culture systems: (a) maturation medium (MM; n = 884), (b) co-cultured with granulosa cells (CG; n = 982) and (c) co-cultured with granulosa cells and cells were further cultured in MM for 12 h (CG + 12hMM; n = 948). Our results showed that ${\alpha}$-tocopherol had no effect on GVBD and MII as compared to control group, but when ${\alpha}$-tocopherol added to maturation medium the rate of cleavage decreased. This indicates interaction of above mentioned factors in any of the treatments showed no significant differences on the rate of maturation and cleavage stages (MII, GVBD and cleavage) (p > 0.05). The oocytes co-cultured with granulosa cells for 24 h had beneficial effects on cleavage rate. The maximum MII and cleavage rates were achieved when oocytes had extra 12 h culture in the maturation medium without granulosa cells. Results also showed our modified co-culture system (CG + 12hMM), improved rates of MII and the cleavage in comparison with other studied maturation systems.