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Purification of the Yolk Protein, and Identification of the Synthetic Site of Its Precursor in Eriocheir japonicus (Decapoda, Brachiura) (동남참게(Eriocheir japonicus)의 난황단백질 정제와 그 전구체의 합성부위 구명)

  • HAN Chang-Hee;BAE Hyun-Hwan
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.25 no.5
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    • pp.432-442
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    • 1992
  • To identify the histological site of synthesis of yolk protein precursor, vitellogenin, by immunocytochemical method in the freshwater crab Eriocheir japonicus, we purified the yolk protein, vitellin, from crude egg extracts, and prepared the anti-rabbit serum against vitellin. Then, the site of vitellogenin synthesis was demonstrated by immunotytochemical method with PAP(peroxidase-antiperoxidase) reaction using the rabbit antiserum aganist vitellin. Female specific serum protein was identified in female serum by immunoelectrophoresis and Ouchterlony's immunodiffusion test for mature male and female sera. Based on the immunoelectrophoresis and Ouchterlony's diffusion test for mature male and female sera and crude egg extracts using antiserum against vitellogenic female serum absorbed with male serum, the female specific serum protein was identified as vitellogenin, detected in female serum only. The major yolk protein, vitellin, was purified from the crude egg extracts by DEAE-cellulose ion exchange chromatography, followed by sepharose CL-4B gel filteration chromatography. The molecular weight of vitellin was estimated to be about 245,000 dalton by sepharose CL-4B gel filteration chromatography. from the results of immunological analysis for vitellin, it was found that the vitellin antiserum contained the antibody against vitellogenin. In the results of immunocytochemical reaction by PAP method with the rabbit antiserum against vitellin, the vitellogenic oocytes and the hepatopancreas of mature female showed positive PAP reaction, but not in follicle cells and previtellogenic oocytes nf ovary, muscle of female and mature male hepatopancreas. Therefore, it showed that the hepatopancreas of mature female is the site of vitellogenic synthesis.

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Study on Nutrient Balance in Paddy Field of Fluvio-Marine Deposit (하해혼성(河海混成) 논토양(土壤)의 양분수지(養分收支)에 관한 연구)

  • Yoo, Chul-Hyun;Yang, Chang-Hyu;Kang, Seung-Weon;Han, Sang-Soo
    • Korean Journal of Soil Science and Fertilizer
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    • v.35 no.5
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    • pp.253-263
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    • 2002
  • To test for the effect of applied fertilizer and nutrients on uptake and transport for paddy rice, two paddy field trials were conducted with Dongjinbyeo in degenerated salt paddy field of Jeonbuk series from 1999 to 2000. After experiment, soil acidity, content of organic matter phosphate, silicate, potassium, calcium, and total nitrogen was increased by application of fresh cattle manure(FCM). Content of Nitrogen in soil layer leached inorganic nitrogen $NO_3$ was higher that that of $NH_4$ and was high in treatment of FCM. Content of $PO_4$ was higher in FCM than other treatments. But content of potassium was in high control. During the growth of rice plant, the amount of water consumption was 477mm. The amount of supplied nitrogen was high in treatment of no nitrogen(NN), 20% reduced application of LCU(LCU-20%), and no fertilizer. In case of phosphate, the supplied amount was more than the consumed amount with the exception of treatment "no phosphate(NP) and no fertilizer(NF)". In case of potassium, the consumed amount was more than the supplied amount in all treatments. The amount of applied nitrogen in the nutrient infiltrated water was high in treatment soil test(ST), C+FCM+Si(Silicate) and the ratio of recovered nitrogen was high in 20% reduced application of LCU. The amount of applied phosphate in the nutrient infiltrated water was high in FCM and that of applied potassium was high in 20% reduced application of LCU. Nitrogen use efficiency of paddy rice was high in 20% reduced application of LCU and use efficiency of phosphate and potassium was high in C+Si(Silicate). Grain yield of rice was high in order of 20% reduced application of LCU>C+Si=C+FCM+Si>C+FCM.

Establishment of Human Embryonic Stem Cells Derived from Frozen-Thawed Blastocysts (동결-융해된 인간 배반포기 배 유래의 배아 간(幹) 세포 배양)

  • Kim, Eun-Young;Nam, Hwa-Kyung;Lee, Keum-Sil;Park, Sae-Young;Park, Eun-Mi;Yoon, Ji-Yeon;Heo, Young-Tae;Cho, Hyun-Jung;Park, Se-Pill;Chung, Kil-Saeng;Lim, Jin-Ho
    • Clinical and Experimental Reproductive Medicine
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    • v.28 no.1
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    • pp.33-40
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    • 2001
  • Objective: This study was to establish the human embryonic stem (ES) cells derived from frozen-thawed blastocyst stage embryo that were destined to be discarded after five years in routine human IVF-ET program. Methods: Frozen-thawed and survived human blastocysts were treated by immunosurgery, and recovered ICM cells were cultured onto STO feeder cell layer and ICM colony was subcultured by mechanical dissociation into clumps. To identify ES cell, alkaline phosphatase staining and expression of Oct4 in replated ICM colonies were examined. Also, to examine the possibility of ES cell differentiation, retinoic acid (RA), basic fibroblast growth factor (b-FGF), nerve growth factor (NGF) were added in culture medium. In addition, to classify the specific cell type, differentiated cells were stained by indirect immunocytochemistry. Results: One ICM colony recovered from frozen-thawed six blastocysts was subcultured, continuously replated during 40 passage culture duration without differentiation. Subcultured colonies were strong positively stained by alkaline phophatase. When the expression of Oct4 in cultured ES colony was examined, Oct4b type is more clearly indicated than Oct4a one although there was not detected in embryoid body or differentiated cells. In differentiated cardiomyocytes from ES colony, cells were beaten regularly (60 times/min). In differentiated neural cells from ES colony, neurofilament (NF) 200 kDa protein, microtubule associated protein (MAP) 2 and ${\beta}$-tubulin of specific marker in neurons, glial fibrillary acidic protein (GFAP) of specific marker in astrocytes and galactocelebrocide (GalC) of specific marker in oligodendrocytes were confirmed by indirect immunocytochemistry. Also, muscle cells were detected by indirect immunocytochemistry. In addition, ES colonies can be successfully cryopreserved. Conclusion: This study suggested that establishment of human ES cells can be successfully derived from frozen-thawed blastocysts that were destined to be discarded, and obtained specific cell types (cardiomyocytes, neurons and muscle cells) through the in vitro differentiation procedures of ES cells.

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Effects of Sargassumpallidum on 2,4,6-Trinitrobenzene Sulfonic Acid-Induced Colitis in Mice (해조가 2,4,6-trinitrobenzene-sulfonic acid로 유발된 염증성 장질환 동물모델에 미치는 영향)

  • Lee, Sang-Wook;Ryu, Bong-Ha;Park, Jae-Woo
    • The Journal of Internal Korean Medicine
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    • v.31 no.2
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    • pp.224-241
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    • 2010
  • Objectives : The aim of the current study was to investigate the effects of Sargassum (Sargassum pallidum (TURN.) C. AG.; SP) on the experimental colitis induced by 2,4,6-trinitrobenzene sulfonic acid (TNBS) in mice. Methods : ICR mice were divided into 7 groups (NOR, CON, $SS50\times5$, $SP20\times3$, $SP50\times3$, $SP20\times5$, $SP50\times5$). TNBS processing was intrarectally applied to all experimental groups on the 3rd experiment day, except the normal group (NOR). For investigating the prophylactic effect, SP at doses of 20 mg/kg ($SP20\times5$) and 50 mg/kg ($SP50\times5$) were orally administered for 5 days. The SP at doses of 20 mg/kg ($SP20\times3$) and 50 mg/kg ($SP50\times3$) were orally administered for 3 days after the colitis induction in order to check the effect of treatment. As a positive control group, sulfasalazine 50 mg/kg ($SS50\times5$) was administrated. Macroscopic findings of epithelial tissue on mice were measured by colon length and macroscopic score. Histologic findings were also checked by crypt cell, epithelial cell, inflammatory cell and edema of submucosa. We measured the ability of SP to inhibit lipid peroxidation and myeloperoxidase activity. We also measured levels of the inflammatory markers, interleukin (IL)-$1\beta$ and cyclooxygenase-2 (COX-2), its transcription factor activation, phospho-NF-${\kappa}B$ (pp65), in the colon by enzyme-linked immunosorbent assay and immunoblot analysis. We measured activation of fecal bacterial enzyme, $\beta$-glucuronidase and degradation activation of fecal glycosaminoglycan (GAG), and hyaluronic acid. Results : Oral administration of SP on mice inhibited TNBS-induced colon shortening and myeloperoxidase activity in the colon of mice as well as IL-$1\beta$ and COX-2 expression. SP also inhibited TNBS-induced lipid peroxidation and pp65 activation in the colon of mice. SP inhibited $\beta$-glucuronidase activation and fecal hyaluronic acid degradation activation as well. Conclusions : SP could be a possible herbal candidate and preventive prebiotic agent for treating inflammatory bowel disease (IBD). Further experiments to differentiate effects of SP on IBD, such as other solutions and extracting times, might be promising.

The Effect of Flooding Time on Ammonia Emission after Application of Liquid Pig Manure in Paddy Soil (돈분뇨 시용 후 담수시기가 암모니아 휘산에 미치는 영향)

  • Lee, Yong-Bok;Lee, Youn;Shin, Pyung-Gyun;Yun, Hong-Bae
    • Korean Journal of Environmental Agriculture
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    • v.30 no.4
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    • pp.377-381
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    • 2011
  • BACKGROUND: Ammonia emissions from field-applied livestock manure are considered a threat to the environment worldwide. In Korea, a large amount of liquid manure was applied in the rice field before rice transplanting in order to reduce chemical fertilizer use. This study was conducted to provide the optimal flooding time after liquid manure application in an attempt to minimize ammonia emission. METHODS AND RESULTS: Ammonia emission from paddy field applied with liquid pig manure following different flooding time was measured using the dynamic chamber method. The five treatments used were : application of liquid pig manure to paddy field in flooding condition (F0T); one day (F1T) and three days (F3T) after flooding; without flooding (NF), and flooding without the application of liquid pig manure (control). Among the treatment, the highest ammonia emission was observed in F0T. The cumulative ammonia emission of F1T and F3T for 12 days were very similar and were about 4.7 times less than that of the F0T treatment. CONCLUSIONS: Ammonia emission in paddy field could be significantly reduced by liquid pig manure application after flooding rather than application of liquid pig manure in flooding condition. Therefore, flooding after liquid pig manure application would provide much more nitrogen for rice growth due to the reduction of ammonia emission.

Immunotoxicological Effects of Aripiprazole: In vivo and In vitro Studies

  • Baek, Kwang-Soo;Ahn, Shinbyoung;Lee, Jaehwi;Kim, Ji Hye;Kim, Han Gyung;Kim, Eunji;Kim, Jun Ho;Sung, Nak Yoon;Yang, Sungjae;Kim, Mi Seon;Hong, Sungyoul;Kim, Jong-Hoon;Cho, Jae Youl
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.365-372
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    • 2015
  • Aripiprazole (ARI) is a commonly prescribed medication used to treat schizophrenia and bipolar disorder. To date, there have been no studies regarding the molecular pathological and immunotoxicological profiling of aripiprazole. Thus, in the present study, we prepared two different formulas of aripiprazole [Free base crystal of aripiprazole (ARPGCB) and cocrystal of aripiprazole (GCB3004)], and explored their effects on the patterns of survival and apoptosis-regulatory proteins under acute toxicity and cytotoxicity test conditions. Furthermore, we also evaluated the modulatory activity of the different formulations on the immunological responses in macrophages primed by various stimulators such as lipopolysaccharide (LPS), pam3CSK, and poly(I:C) via toll-like receptor 4 (TLR4), TLR2, and TLR3 pathways, respectively. In liver, both ARPGCB and GCB3004 produced similar toxicity profiles. In particular, these two formulas exhibited similar phospho-protein profiling of p65/nuclear factor $(NF)-{\kappa}B$, c-Jun/activator protein (AP)-1, ERK, JNK, p38, caspase 3, and bcl-2 in brain. In contrast, the patterns of these phospho-proteins were variable in other tissues. Moreover, these two formulas did not exhibit any cytotoxicity in C6 glioma cells. Finally, the two formulations at available in vivo concentrations did not block nitric oxide (NO) production from activated macrophage-like RAW264.7 cells stimulated with LPS, pam3CSK, or poly(I:C), nor did they alter the morphological changes of the activated macrophages. Taken together, our present work, as a comparative study of two different formulas of aripiprazole, suggests that these two formulas can be used to achieve similar functional activation of brain proteins related to cell survival and apoptosis and immunotoxicological activities of macrophages.

Growth Properties of Mixtures with Mixed Organic Fertilizer and Dried Food Waste Powder in Pakchoi (Brassica rapa L.) (음식물류폐기물건조분말과 혼합유기질비료 혼합물의 청경채 생장 효과)

  • Kim, Young-Sun;Cho, Sung-Hyun;Lee, Hoonsoo;Lee, Geung-Joo
    • Journal of the Korea Organic Resources Recycling Association
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    • v.29 no.3
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    • pp.17-25
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    • 2021
  • This study was conducted to investigate effects of mixture with dried food waste powder (FWP) and mixed organic fertilizer (MOF) on growth of pakchoi. As compared with non-fertilizer treatment (NF) or control (MOF treatment), growth of pakchoi in FWP treatments (2,500 kg/ha, 5,000 kg/ha, 10,000 kg/ha) was inhibited by salt (NaCl) content in the FWP. In comparison with control, mixtures of MOF and FWP (FWP10, FWP20, and FWP30 treatment) were not significantly different, and their salt content correlated with pakchoi growth factors negatively (P<0.05). Applied of FWP10, (FWP10: 2,500 kg/ha, 2FWP10: 5,000 kg/ha, 3FWP10: 7,500 kg/ha, 4FWP10: 10,000 kg/ha), growth factors of FWP10, 2FWP10 and 3FWP10 treatment were not significantly different than those of chemical fertilizer treatment, and of 4FWP10 decreased. Correlation coefficient between NaCl supply by FWP10 application and growth factor was negative (P<0.01). These results indicated that FWP was used as another source of organic fertilizer, and the organic fertilizers blending with FWP inhibited a pakchoi growth by increase of salt content containing in the them or of salt supplying amount after their application.

Impacts of Different Organic Fertilizers on Soil Fertility and Soil Respiration for a Corn (Zea mays L.) Cropping System (옥수수 밭에서 유기질 비료가 토양 비옥도 및 토양 호흡에 미치는 영향)

  • Mavis, Brempong Badu;Hwang, Hyun Young;Lee, Sang Min;Lee, Cho Rong;An, Nan Hee
    • Journal of the Korea Organic Resources Recycling Association
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    • v.30 no.4
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    • pp.151-163
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    • 2022
  • This study was conducted to promote organic fertilizer(s) that sustain soil productivity for corn production and protect the environment as required by the Act on the promotion of eco-friendly agriculture. It was conducted at the research station of the Organic Agriculture Division of the National Institute of Agricultural. The treatments consisted of Compost (Com), Bokashi as fermented organic fertilizer (FOF), and mixed expeller pressed cake (PC). They were applied at 174 kg N /ha to field corn, together with a 'no fertilizer' check in Randomized Complete Block Design. At eight weeks after transplanting (WAT) corn, compost increased soil carbon (C) and nitrogen (N) to 7.48 and 0.76 g/kg respectively, while other fertilizers maintained the initial levels (before treatment application). At corn harvest (13 WAT), soil chemical properties (total C, total N, pH, electrical conductivity, P2O5, Ca, K, and Mg) were similar among all organic fertilizer treatments. For soil respiration, FOF increased soil CO2 respiration by 31-76% above other fertilizer treatments. However, there were no prominent changes in the trends of CH4 fluxes following the two mechanical weeding operations. Fermented organic fertilizer affected N2O emissions between 87-96% lower than other fertilizer treatments. Compared to the initial microbial densities, FOF increased fungi and actinomycete colony foming unit by 25 and 16% at harvest. Therefore, the additional potential of improving soil biological fertility and local availability of raw materials make FOF a better option to sustain soil productivity while protecting the environment.

Diversity and Antimicrobial Activity of Actinomycetes Isolated from Rhizosphere of Rice (Oryza sativa L.) (벼 근권에서 분리한 방선균의 다양성과 항균 활성)

  • Lee, Hye-Won;Ahn, Jae-Hyung;Weon, Hang-Yeon;Song, Jaekyeong;Kim, Byung-Yong
    • The Korean Journal of Pesticide Science
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    • v.17 no.4
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    • pp.371-378
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    • 2013
  • Various microorganisms live in soil, of which those colonizing rhizosphere interact with nearby plants and tend to develop unique microbial communities. In this study, we isolated diverse actinomycetes from rhizosphere of rice (Oryza sativa L.) cultivated in fertilized (APK) and non-fertilized (NF) paddy soils, and investigated the diversity and antimicrobial activity of them. Using four kinds of selective media, 152 isolates were obtained from the soil samples and identified by determining 16S rRNA gene sequence. All of the isolates showed 99.0%~100.0% similarities with type strains and were classified into six genera: Dactylosporangium, Micromonospora, Kitasatospora, Promicromonospora, Streptomyces and Streptosporangium. Most of the isolates, 143 isolates, were classified into the genus Streptomyces. Additionally, many isolates had antimicrobial activity against plant pathogens, especially Magnaporthe oryzae (rice blast pathogen) in fungi. These findings demonstrated that rice rhizosphere can be a rich source of antagonistic actinomycetes producing diverse bioactive compounds.

Effects of Nicotine, Cotinine and Benzopyrene as Smoke Components on the Expression of Antioxidants in Human Bronchial Epithelial Cells (흡연성분 중 Nicotine, Cotinine, Benzopyrene이 인체 기관지 상피세포에서 항산화제의 발현에 미치는 영향)

  • Kim, Yong Seok;Lee, Jae Hyung;Kim, Sang Heon;Kim, Tae Hyung;Sohn, Jang Won;Yoon, Ho Joo;Park, Sung Soo;Shin, Dong Ho
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.3
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    • pp.197-202
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    • 2007
  • Background: Cigarette smoking is an important risk factor for chronic bronchitis and COPD. Airway epithelial cells exposed to cigarette smoke components such as nicotine, cotinine and benzopyrene can generate reactive oxygen species (ROS) and be subject to oxidative stress. This oxidative stress can induce the inflammatory response in the lung by the oxidant itself or by the release of proinflammatory cytokines. It has been reported that nicotine stimulates ROS, which are associated with NF-${\kappa}B$. Methods: Beas2B cells were treated with nicotine, cotinine and benzopyrene. RT PCR was used to measure the expression of several antioxidant factors using the total RNA from the Beas2B cells. The level of superoxide dismutase(CuZnSOD), thioredoxin, glutathione reductase expression was examined. Results: 0.5 to 4 hours after the benzopyrene, nicotine and cotinine theatments, the level of thioredoxin and glutathione reductase expression decreased. Longer exposure to these compounds for 24 to 72 hours inhibited the expression of most of these antioxidant factors. Conclusion: During exposure to smoke compounds, thioredoxin and glutathione reductase are the key antioxidant factors induced sensitively between 0.5 and 4 hours but the levels these antioxidants decrease between 24 hour and 72hours.