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Application Effects of Fermented Mixed Organic Fertilizer Utilizing By-Products on Yield of Chinese Cabbage and Soil Environment (부산물 활용 발효 유기질비료가 배추 수량 및 토양환경에 미치는 영향)

  • An, Nan-Hee;Lee, Sang-Min;Oh, Eun-mi;Lee, Cho-Rong;Gong, Min-Jae
    • Journal of the Korea Organic Resources Recycling Association
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    • v.28 no.4
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    • pp.77-85
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    • 2020
  • This study analyzes the effects of mixed fermented organic fertilizer on chinese cabbage growth and soil properties in order to investigate the nutritional effects of organic fertilizers, which are developed as an alternative fertilizer for imported castor oil cake. In this study, four treatments were set up: 100% and 200% rate of nitrogen application (320 kg ha-1 for Chinese cabbage) on mixed fermented organic fertilizer A(FA) and mixed fermented organic fertilizer B(FB), respectively, 100% rates of the mixed expeller cake (MEC) fertilizer, and the untreated control. Results revealed that the growth and yield of Chinese cabbage increased as more fermented organic fertilizer was used. However, while there were no significant differences in growth characteristics between treatments of 100% rate of mixed fermented organic fertilizer and 100% rate of MEC, the impacts on yields resulted similar. The nitrogen use efficiency (NUE) of Chinese cabbage was measured a range of 20-31% depending on the response to treatment. The 100% FA showed the same as NUE and nitrogen absorption with 100% rate of MEC. Regarding soil properties after cultivation, there were no significant differences among the effects of fertilizers in pH, EC, soil organic matter, and available phosphate. However, the content of exchangeable cations(K, Ca, Mg) was higher in areas treated with mixed fermented organic fertilizer than in untreated areas. Furthermore, the bacterial population density in the soil was higher in areas treated with mixed fermented organic fertilizer than in untreated areas and increased as more mixed fermented organic fertilizer was used. There were no significant differences in the population density of actinomycetes and fungi when fertilizer was applied to the soil. These results also show that FA, as a alternative organic fertilizer for imported castor oil cake, has similar nutritional effects as that of MEC. Therefore, further research the appropriate amounts of fertilizer is required to achieve economical and eco-friendly nutrient management.

LC-MS/MS analysis and anti-inflammatory effects of crude extract from Coptidis Rhizoma (황련 추출물의 LC-MS/MS 분석 및 항염증 효과)

  • Min-Jung, Kim;Ye-Jin, Yang;Kwang-Youn, Kim;Hun Hwan, Kim;Jae Dong, Son;Ju-Hye, Yang;Dong bin, Lee;Woo Hyun, Kim;Hu-Jang, Lee;Seon Been, Bak;Kwang-Il, Park
    • Herbal Formula Science
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    • v.31 no.1
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    • pp.1-10
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    • 2023
  • Objectives : The main aim of this study was to examine the LC-MS/MS used to identify phenolic compounds of CRE(Coptidis Rhizoma 70% EtOH Extract). Also, we investigated antioxidative activities and Anti-inflammatory activities. Methods : LC-MS/MS Analysis HPLC and LC-MS/MS were performed on a 1260 series HPLC system (Agilent Technologies, Inc., California, USA) and 3200 QTrap tandem mass system (Sciex LLC) operated in positive ion mode (spray voltage set at -4.5 kV). The solvent used was DW and Acetonitrile containing 0.1% formic acid, a gradient system was used at a flow rate of 0.5 mL/min for analysis, and a Prontosil C18 column (length, 250 mm; inner diameter, 4.6 mm; particle size, 5 ㎛; Phenomenex Co., Ltd., California, USA, Biochoff Chromatography) was used. The solvent conditions used in the mobile phases were 0-10 min at 10-15% B, 10-20 min at 20% B, 20-30 min at 25%, 30-40 min at 40%, 40-50 min at 70%, 50-60 min at 95%, and 60-70 min at 95%. The analysis was performed at a wavelength of 284 nm and a temperature of 35℃. The cell viability was measured using a 3-(4,5-dimethyethiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity. We examined the effects of CRE on the lipopolysaccharide (LPS)-induced production of nitric oxide (NO) in a RAW 264.7 cells Results : The chemical analysis CRE by Liquid chromatography-tandem mass spectrometry (LC-MS/MS) confirmed that Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid as phenolic components. DPPH radical scavenging activity was the inhibitory activity of CRE showed at 200 ㎍/mL a statistically significant level. MTT assay demonstrated that the CRE did not have a cytotoxic effect in RAW 264.7 and LPS-induced RAW264.7 cells. Also, CRE reduced NO production in RAW 264.7 cells stimulated with LPS. Conclusions : Based on these findings, The chemical analysis 4 major components CRE such as Rosmarinic acid, Ferrulic acid, 3-O-feruloylquinic acid, and 5-O-feruloylquinic acid. Moreover, we confirmed that CRE has effects antioxidant and anti-inflammatory. The results demonstrate that CRE can be used as an antioxidant and a powerful chemopreventive ingredient against inflammatory diseases.

Effects of Feeding Concentrate on Development of Intestinal Tract and Composition of Muscle in Hanwoo Calves before Weaning (이유 전 농후사료(무조사료) 급여가 한우 송아지의 장관발달 및 근육 성분에 미치는 영향)

  • Kim, W.Y.;Lee, S.H.;Kim, S.K.;Lee, J.H.;Nho, W.G.;Hwang, J.H.;Lee, S.S.;Yeo, J.M.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.15 no.1
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    • pp.123-130
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    • 2013
  • The present study was conducted to investigate effects of feeding concentrate on development of intestinal tract and composition of muscle in suckling Hanwoo calves before weaning(60 days of age). Twenty-four Hanwoo calves(12 heifers and 12 bulls) at six days of age were randomly assigned to one of three dietary treatments[hay and two levels(low and high) of concentrate feeding] and given each diet with free access to their dams for suckling until 60 days of age. At 60 days of age, two calves from each treatment were used for post-mortem examination of intestine development and chemical analysis of muscle. Feed intake between 31 and 60 days of age was significantly(P<0.05) higher for the high concentrate treatment(532.4g/d) than for other treatments(78.9 and 108.9g/d for hay and low concentrate treatments, respectively). Although feed intake showed a large difference between the low and high concentrate treatments, body weights at birth and 60 days of age were similar between the treatments, suggesting that calves in the low concentrate feeding were supplied more milk from their dams than those in the high concentrate feeding. The ratio of intestine weight to body weight at 60 days of age was increased in the high concentrate(3.35%), compared with those in the hay(2.93%) and low concentrate(3.03%) treatments. The chemical composition of muscle in sirloin and hind leg was not affected by the treatments. In conclusions, the results of the present study showed that feeding concentrate to Hanwoo calves before weaning might stimulate intestine development relative to feeding hay.

Development of the paper bagging machine for grapes (휴대용 포도자동결속기 개발연구)

  • Park, K.H.;Lee, Y.C.;Moon, B.W.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.11 no.1
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    • pp.79-94
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    • 2009
  • The research project was conducted to develop a paper bagging machine for grape. This technology was aimed to highly reduce a labor for paper bagging in grape and bakery. In agriculture labor and farm population has rapidly decreased since 1980 in Korea so there was so limit in labor. In particular there is highly population in women and old age at rural area and thus labor cost is so high. Therefore a labor saving technology in agricultural sector might be needed to be replaced these old age with mechanical and labor saving tool in agriculture. The following was summarized of the research results for development of a paper bagging machine for grape. 1. Development of a new paper bagging machine for grape - This machine was designed by CATIA VI2/AUTO CAD2000 programme. - A paper bagging machine was mechanically binded a paper bag of grape which should be light and small size. This machine would be designed for women and old age with convenience during bagging work at the field site. - This machine was manufactured with total weight of less than 350g. - An overage bagging operation was more than 99% at the actual field process. - A paper bagging machine was designed with cartridge type which would be easily operated between rows and grape branches under field condition. - The type of cartridge pin was designed as a C-ring type with the length of 500mm which was good for bagging both grape and bakery. - In particular this machine was developed to easily operated among vines of the grape trees. 2. Field trials of a paper bagging machine in grape - There was high in grape quality as compared to the untreated control at the application of paper bagging machine. - The efficiency of paper bagging machine was 102% which was alternative tool for the conventional. - The roll pin of paper bagging machine was good with 5.3cm in terms of bagging precision. - There was no in grape quality between the paper bagging machine and the conventional method. - Disease infection and grape break was not in difference both treatments.

Studies on the Evaluation of the Spent Composts of Selenium-Enriched Mushrooms as a Feed Selenium Source (셀레늄강화 버섯폐배지에 대한 사료 셀레늄공급원으로의 평가 연구)

  • Kim, W.Y.;Min, J.K.
    • Journal of Practical Agriculture & Fisheries Research
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    • v.7 no.1
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    • pp.118-130
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    • 2005
  • This study was conducted to evaluate the spent composts of selenium-enriched mushrooms as a feed selenium Source. Total selenium (Se) contents and Se profiles in the spent mushroom composts (SMC) were determined. In addtion, we also investigated the metabolism in relation to Se accumulation in the mushroom. Mushrooms used in this study were Flammulina velutipes and Se enriched mushrooms were grown for 60 days by adding 2 mg of inorganic Se (Na2SeO3) per kg of mushroom composts (MC) on as-fed basis and it was compared with mushrooms not to add Se to the MC. Total Se contents for Se-treated mushrooms were significantly increased (P<0.0001) by 20-fold (4.51 ㎍/g of dry) compared to Se-untreated (0.23 ㎍/g of dry). On the contrary, organic Se proportion was significantly lower (P<0.0001) in the Se-treated mushroom (72.3%) than Se-untreated (100%, not analytically detected of inorganic Se). Se distribution upon a length in the Se-treated mushrooms was the highest in the bottom part (6.86 ㎍/g of dry) near to MC, and top and middle parts were significantly lower (3.71 and 3.01 ㎍/g of dry, respectively) than the bottom (P<0.001). In the SMC from Se-treated mushrooms, a high concentration of Se (5.04 ㎍/g of dry) was still remained, but that from Se-untreated mushrooms was significantly low (P<0.0001) as 0.08 ㎍/g of dry. Se-treated SMC showed a high rate of organic Se (65.67%), suggesting that most of inorganic Se in the SMC was converted to organic Se by mushroom mycelia, and Se-untreated SMC showed 100% of organic Se, not being detected of inorganic Se. Prior to mycelia inoculation in the mushroom culture, the sterilization of MC brought approximately 18% of Se loss in the MC. This result is in accordance with facts generally known that Se is weak in the high temperature and it is consequently volatilized under that condition. Apparent and net accumulation rates (%) for Se into mushrooms were 14.81 and 10.14%, respectively and their difference (4.67%) is considered that it is due to the volatilization into the air via metabolic process of mushroom itself. From the result of this study, inorganic Se addition to MC for mushroom improved the Se content in the mushroom and SMC from Se-enriched mushrooms contained a high concentration of Se. Mycelium and fruiting body from mushrooms converted inorganic Se in MC to organic Se, indicating a high proportion of organic Se in the mushroom and SMC. Therefore, Se in Se-enriched mushroom and SMC was recognized as Se sources of food for human as well as feed for livestock.

Correlation between litter decomposition rate of Quercus mongolica leaf and microclimatic factors at Mt. Jeombongsan (점봉산 신갈나무 낙엽의 분해율과 미기상요인과의 상관관계 분석)

  • Ho-Yeon Won;Young-Sang Lee;Jae-Seok Lee;Il-Hwan Lee
    • Korean Journal of Environmental Biology
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    • v.40 no.4
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    • pp.455-463
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    • 2022
  • To understand functional changes of forest ecosystems due to climate change, correlation between decomposition rate of leaf litter, an important function of forest ecosystems, and microclimatic factors was analyzed. After 48 months elapsed, percent remaining weight of Quercus mongolica leaf litter was 27.1% in the east aspect and 37.0% in the west aspects. Decay constant of Q. mongolica leaf litter was 0.33 in the east aspect and 0.25 in the west aspect after 48 months elapsed. Initial C/N ratio of Q. mongolica leaf litter was 38.5. After 48 months elapsed, C/N ratio of decomposing Q. mongolica leaf litter decreased to 13.43 in the east aspect and 16.72 in the west aspect. Average air temperature and soil temperature during the investigation period of the research site were 8.2±9.0 and 9.1±9.3 in the east and 8.5±7.4 and 9.3±7.3℃ in the west aspect, respectively, with the west aspect showing higher air and soil temperatures. Soil moisture showed no significant difference between east and west aspects (average soil moisture: 19.4±11.0% vs. 20.5±5.7%). However, as a result of analyzing the correlation between decomposition rate and microclimatic factors, it was found that the decomposition rate and soil moisture has a positive correlation(r=0.426) in the east aspect but not in the west aspect. Our study shows that the correlation between decomposition rate and microclimatic factors can be significantly different depending on the direction of the aspect.

Effect of Fusion Procedure on the Development of Embryos Produced by Somatic Cell Nuclear Transfer in Hanwoo (Korean Cattle) (한우에서 융합방법이 체세포 핵이식 수정란의 발달에 미치는 영향)

  • Im, G.S.;Yang, B.S.;Park, S.J.;Chang, W.K.;Park, C.S.
    • Korean Journal of Animal Reproduction
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    • v.24 no.4
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    • pp.365-373
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    • 2000
  • The purpose of this study was to investigate the effects of the fusion pulses and fusion media on fusion rate and the development of embryos produced by somatic cell nuclear transfer in Hanwoo (Korean cattle). Nuclear donor cumulus and fetal fibroblast cells were cultured in Dulbecco's modified Eagle medium supplemented with 10% fetal bovine serum at 38.5$^{\circ}C$ in a humidified atmosphere of 5% $CO_2$in air. The in vitro matured oocytes were enucleated and then the isolated donor cells were introduced. The cumulus cell and cytoplast were fused using one pulse of 70 volts for 40$mutextrm{s}$, two pulses of 70 volts for 40$mutextrm{s}$ and one pulse of 180 volts for 15$mutextrm{s}$. The fetal fibroblast cell and cytoplast were fused using one pulse of 180 volts for 15$mutextrm{s}$ or 30$mutextrm{s}$. The cumulus cell and cytoplast were fused using mannitol and Zimmerman cell fusion medium (ZCFM) as a fusion medium. The fused embryos were activated after the fusion with 10 $\mu$M calcium ionophore for 5 min and 2 mM 6-dimethyl- aminopurine for 3 h. The nuclear transfer embryos were cultured in 500 ${mu}ell$ well of modified CR1aa supplemented with 3 mg/$m\ell$ BSA in th $\varepsilon$ four well dish cove red with mineral oil. After 3 days culture, culture medium was changed into modified CRlaa medium containing 1.5 mg/$m\ell$ BSA and 5% FBS for 4 days. The incubation environment was 5% $CO_2$, 5% $O_2$, 90% $N_2$ at 38.5$^{\circ}C$. When the cumulus cells were fused with enucleated oocytes by three different fusion pulses, one pulse of 180 volts for 15 $mutextrm{s}$ yielded the highest fusion rate and developmental rate to blastocyst among the pulses (P<0.05). When the fetal fibroblast cells were fused with enucleated oocytes, one pulse of 180 volts for 30$mutextrm{s}$ yielded significantly higher fusion rate compared with that for 15 $mutextrm{s}$(P<0.05). The present result indicates that the fusion rate between karyoplast and cytoplast was affected by the cell type and the optimal fusion condition was different according to cell type or size. When the fusion was conducted by the use of mannitol and ZCFM, the fusion rate was 71.2% and 65.8%, respectively. The developmental rates to blastocyst were 37.8% and 39.8%, respectively. There was no significant difference between two fusion media in the developmental rate of cumulus cell nuclear transfer embryos. These results indicate that optimal electric current should be selected according to cell type.

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Introduction of the Best Practices in the Pakistan Gulpur HEPP (파키스탄 Gulpur 수력발전 현장의 Best Practices 소개)

  • JANG, Ock Jae;HONG, Won Pyo;CHAE, Hee Moon
    • Proceedings of the Korea Water Resources Association Conference
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    • 2022.05a
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    • pp.216-217
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    • 2022
  • Gulpur 수력발전 프로젝트는 전력난을 겪고 있는 파키스탄에 102 MW 규모의 수력발전소를 건설하여 30년 동안 운영 관리한 후 파키스탄 정부로 양도하는 IPP(Independent Power Producing) 형식의 투자사업이다. 남동발전과 DL E&C, 롯데건설이 Sponsor로서 출자한 자본금과, ADB, IFC, K-EXIM 등의 대주단로부터의 차입금을 재원으로 하여 소요 사업비를 조달하고 사업을 개발하였다. DL E&C와 롯데건설이 EPC(Engineering, Procurement, Construction)를 수행하였고, 이산이 Design consultant의 역할을 수행하였다. Gulpur 수력발전 프로젝트의 발전형식은 수로식(run-of-river)으로 201 m3/s의 발전유량과 102 MW의 발전 시설용량을 이용하여 연평균예상발전량은 398 GWh이다. 주요 구조물로는 설계 재현빈도 1년의 유수전환시설(가물막이댐 & 가배수터널)과 콘크리트 중력식댐(H 67 m, L 205 m), 도수터널(D 6.7 m, L 215 m, 2기), 옥외형 발전소 (H 51 m, W 60 m, L 38 m, Kaplan 2기)가 있으며, 2015년 10월 착공하여 2020년 3월 상업발전을 시작하였다. 본 프로젝트는 DL E&C의 첫 번째 EPC 해외수력발전 프로젝트이다. 따라서 프로젝트의 성공적 수행을 위한 경제적 설계, 시공의 효율성 및 안정성 확보 등을 위하여 많은 연구를 수행하는 과정에서 다양한 기술 개선을 이룰 수 있었다. 본고에서는 Gulpur 프로젝트를 통하여 도출된 성공 사례들을 소개 및 공유하고자 한다. 첫 번째로 콘크리트 중력식댐 시공을 위한 유수전환시설의 최적 설계빈도를 산정하였다. 일반적으로 유수전환시설의 규모는 설계기준에 제시된 설계 재현빈도를 이용하는데, 해외 설계기준에서는 10년, 국내 설계기준에서는 1~2년으로 다르게 제시되어 있는 문제점이 있다. 유수전환시설의 규모는 프로젝트의 경제성에 큰 영향을 미치기 때문에 최적 설계빈도의 결정이 필요하며, 위험도분석기법(Risk Analysis)과 기대화폐가치법(Expected Monetary Value)을 이용하여 유수전환시설의 최적 설계 재현빈도와 이에 영향을 미치는 인자를 분석하였다. 위험도는 몬테카를로 시뮬레이션으로 산정된 가물막이댐 파괴확률과 재현빈도를 이용하여 산정된 가물막이댐 월류확률을 고려하였으며, 비용 및 피해액으로는 유수전환시설의 공사비, 가물막이댐 파괴시의 재건설비용과 지체보상금, 가물막이댐 월류시의 복구비용을 고려하였다. 이에 대한 연구결과로, 유수전환시설의 사용기간과 월류시의 복구비용이 유수전환시설의 설계 재현기간 결정에 가장 큰 영향을 미치는 것으로 나타났고, 특히 월류시의 복구비용이 작을수록 낮은 설계 재현빈도를 선택하는 것이 타당한 것으로 나타났다. 예를 들어, 유수전환시설의 사용기간이 3 ~ 5년, 복구비용이 0.5 ~ 1.0 mil USD 이하인 조건에서 가물막이시설의 최적 설계빈도는 1년 ~ 2년인 것으로 나타났다. 또한, 유수전환시설의 사용기간은 본댐의 규모와 시공기간 등을 고려하여 결정되는 사항으로 설계자가 임의 조정할 수 없지만, 복구비용은 시공 관리자에 따라 결정되는 부분으로, 적극적 홍수 피해 저감 및 복구방안을 마련하는 것이 프로젝트의 경제성을 향상시킬 수 있다는 것을 알 수 있었다. 두 번째로 프로젝트의 경제성 향상, 홍수기 댐 시공시의 안전성 확보를 위하여 홍수 조기경보시스템(Early Warning System)을 개발 및 활용하였다. 수로식(Run-of-river) 수력발전댐은 대부분 산악지역에 위치하기 때문에 국지성 강우 및 급한 지형 경사로 인하여 돌발홍수(flash flood)의 발생 가능성이 높다. 따라서 시공 중 홍수(월류) 발생을 미리 감지하고 현장에 전파할 수 있는, 수로식(Run-of-river) 수력발전댐 현장을 위한 홍수 조기경보시스템이 필요하며, 이를 리스크 인식, 모니터링 및 경보, 전파 및 연락, 반응 능력 향상의 4가지 부분으로 나누어 구축하였다. 리스크 인식 부분에서는 가물막이댐 월류 발생 상황에 대한 위험도, 취약성, 리스크를 제시하였으며, 모니터링 및 경보 부분에서는 상류 측정수위에서 유도된 현장 예상수위와 실제 현장 측정 수위를 대상으로 경보홍수위와 위험홍수위로 나누어 관리하였다. 전파 및 연락 부분에서는 현장 시공 조직을 활용하여 홍수시를 대비한 비상연락체계도(Emergency communication flow chart)를 운영하였으며, 반응 능력 향상을 위해 비상연락체계도의 팀별 Action plan을 상세화 하였다. 세 번째로 현장의 지질특성과 50여 차례 발파시험으로 현장 고유의 발파진동감쇄곡선을 도출하였으며, 이를 통해 현장의 시공성과 콘크리트 품질 확보를 동시에 달성할 수 있는 방안을 제시하였다. 콘크리트댐 공사에서는 제한된 공기 내에 공사를 완료하기 위해 사면부 굴착과 콘크리트 타설이 동시에 수행될 수밖에 없는 문제점을 가지고 있다. 그러나 신규 콘크리트 타설면 근처에서 발파를 수행하는 경우 발파로 발생되는 탄성파가 일정 수준을 초과하게 되면, 콘크리트 양생에 영향을 주게 된다. 따라서 다수의 현장 발파시험을 통해 발파거리와 최대진동속도의 상관관계 즉, 발파진동감쇄곡선을 도출함으로써 현장의 발파진동특성을 도출할 수 있었다. 또한, 기존 연구 논문들을 통해 콘크리트 재령기간 별 안전진동속도를 선정하고, 해당 안전진동속도를 초과하지 않는 범위에서 콘크리트 타설면과 발파위치의 거리에 따라 1회 발파 가능한 장약량을 산정하여 적용하였다. 이와 같은 체계적인 접근을 통해 콘크리트 타설과 발파 작업 동시 수행에 대한 논란을 해소할 수 있었다.

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Quality characteristics of different parts of garlic sprouts produced by smart farms during growth (스마트팜 생산 새싹마늘의 부위별 및 생육 기간에 따른 품질 특성)

  • Yu-Ri Choi;Su-Hwan Kim;Chae-Mi Lee;Dong-Hun Lee;Chae-Yun Lee;Hyeong-Woo Jo;Jae-Hee Jeong;Imkyung Oh;Ho-Kyung Ha;Jungsil Kim;Chang-Ki Huh
    • Food Science and Preservation
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    • v.30 no.2
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    • pp.272-286
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    • 2023
  • Garlic sprouts can provide data on functional and food processing materials. This study compared the leaves, bulbs, and roots of garlic sprouts grown on smart farms during two growth periods (20 and 25 days). In addition, data for garlic bulbs grown in open fields were presented as reference materials. All garlic sprouts' total free sugar content decreased as the growth period increased. All plant parts' total organic acid content decreased as the growth period progressed, except for the root section. Potassium, phosphorus, and sulfur content increased during growth in all parts of the garlic sprouts. Alliin content decreased in all parts of the plant over time, whereas thiosulfinate content increased in the roots but decreased in the leaves and bulbs. Total polyphenol content increased in all parts of the plant during the growth period, except for the bulb, whereas the flavonoid content did not change significantly over time. The 2,2-diphenyl-1-picrylhydrazy (DPPH) and 2,2'-azinobis (3-ethylben-zothiazoline 6-sulfonate) (ABTS) free radical scavenging activities, as well as the superoxide dismutase (SOD)-like activity of garlic sprouts were 37.45-65.47%, 59.12-89.81%, and 89.52-98.59%, respectively. These activities tend to decrease during the growth period. Here, we showed that garlic sprouts have higher levels of functional substances and physiological activities than general garlic sprouts. It was also determined that a growth period of 20 days was suitable for garlic sprouts. Data for research on functional and food-processing materials can be obtained by analyzing garlic sprouts produced by smart farms.

Establishment of Test Conditions and Interlaboratory Comparison Study of Neuro-2a Assay for Saxitoxin Detection (Saxitoxin 검출을 위한 Neuro-2a 시험법 조건 확립 및 실험실 간 변동성 비교 연구)

  • Youngjin Kim;Jooree Seo;Jun Kim;Jeong-In Park;Jong Hee Kim;Hyun Park;Young-Seok Han;Youn-Jung Kim
    • Journal of Marine Life Science
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    • v.9 no.1
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    • pp.9-21
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    • 2024
  • Paralytic shellfish poisoning (PSP) including Saxitoxin (STX) is caused by harmful algae, and poisoning occurs when the contaminated seafood is consumed. The mouse bioassay (MBA), a standard test method for detecting PSP, is being sanctioned in many countries due to its low detection limit and the animal concerns. An alternative to the MBA is the Neuro-2a cell-based assay. This study aimed to establish various test conditions for Neuro-2a assay, including cell density, culture conditions, and STX treatment conditions, to suit the domestic laboratory environment. As a result, the initial cell density was set to 40,000 cells/well and the incubation time to 24 hours. Additionally, the concentration of Ouabain and Veratridine (O/V) was set to 500/50 μM, at which most cells died. In this study, we identified eight concentrations of STX, ranging from 368 to 47,056 fg/μl, which produced an S-shaped dose-response curve when treated with O/V. Through inter-laboratory variability comparison of the Neuro-2a assay, we established five Quality Control Criteria to verify the appropriateness of the experiments and six Data Criteria (Top and Bottom OD, EC50, EC20, Hill slop, and R2 of graph) to determine the reliability of the experimental data. The Neuro-2a assay conducted under the established conditions showed an EC50 value of approximately 1,800~3,500 fg/μl. The intra- & inter-lab variability comparison results showed that the coefficients of variation (CVs) for the Quality Control and Data values ranged from 1.98% to 29.15%, confirming the reproducibility of the experiments. This study presented Quality Control Criteria and Data Criteria to assess the appropriateness of the experiments and confirmed the excellent repeatability and reproducibility of the Neuro-2a assay. To apply the Neuro-2a assay as an alternative method for detecting PSP in domestic seafood, it is essential to establish a toxin extraction method from seafood and toxin quantification methods, and perform correlation analysis with MBA and instrumental analysis methods.