• Journal of Microbiology
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• 제45권2호
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• pp.158-167
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• 2007

In this study, we have cloned a novel cDNA encoding for a papain-family cysteine protease from the Uni-ZAP XR cDNA library of the polychaete, Periserrula leucophryna. This gene was expressed in Escherichia coli using the T7 promoter system, and the protease was characterized after partial purification. First, the partial DNA fragment (498 bp) was amplified from the total RNA via RT-PCR using degenerated primers derived from the conserved region of cysteine protease. The full-length cDNA of cysteine protease (PLCP) was prepared via the screening of the Uni-ZAP XR cDNA library using the $^{32}P-labeled$ partial DNA fragment. As a result, the PLCP gene was determined to consist of a 2591 bp nucleotide sequence (CDS: 173-1024 bp) which encodes for a 283-amino acid polypeptide, which is itself composed of an 59-residue signal sequence, a 6-residue propeptide, a 218-residue mature protein, and a long 3'-noncoding region encompassing 1564 bp. The predicted molecular weights of the preproprotein and the mature protein were calculated as 31.8 kDa and 25 kDa, respectively. The results of sequence analysis and alignment revealed a significant degree of sequence similarity with other eukaryotic cysteine proteases, including the conserved catalytic triad of the $Cys^{90},\;His^{226},\;and\;Asn^{250}$ residues which characterize the C1 family of papain-like cysteine protease. The nucleotide and amino acid sequences of the novel gene were deposited into the GenBank database under the accession numbers, AY390282 and AAR27011, respectively. The results of Northern blot analysis revealed the 2.5 kb size of the transcript and ubiquitous expression throughout the entirety of the body, head, gut, and skin, which suggested that the PLCP may be grouped within the cathepsin F-like proteases. The region encoding for the mature form of the protease was then subcloned into the pT7-7 expression vector following PCR amplification using the designed primers, including the initiation and termination codons. The recombinant cysteine proteases were generated in a range of 6.3 % to 12.5 % of the total cell proteins in the E. coli BL21(DE3) strain for 8 transformants. The results of SDS-PAGE and Western blot analysis indicated that a cysteine protease of approximately 25 kDa (mature form) was generated. The optimal pH and temperature of the enzyme were determined to be approximately 9.5 and $35^{\circ}C$, respectively, thereby indicating that the cysteine protease is a member of the alkaline protease group. The evaluation of substrate specificity indicated that the purified protease was more active towards Arg-X or Lys-X and did not efficiently cleave the substrates with non-polar amino acids at the P1 site. The PLCP evidenced fibrinolytic activity on the plasminogen-free fibrin plate test.

• Asian Pacific Journal of Cancer Prevention
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• 제15권19호
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• pp.8183-8189
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• 2014

Background: A comparative cross-sectional study involving oral cancer patients and healthy individuals was designed to investigate associations between retinol, ${\alpha}$-tocopherol and ${\beta}$-carotene with the risk of oral cancer. Materials and Methods: This study included a total of 240 matched cases and controls where subjects were selected from the Malaysian Oral Cancer Database and Tissue Bank System (MOCDTBS). Retinol, ${\alpha}$-tocopherol and ${\beta}$-carotene levels and intake were examined by high-performance liquid chromatography (HPLC) and food frequency questionnaire (FFQ) respectively. Results: It was found that results from the two methods applied did not correlate, so that further analysis was done using the HPLC method utilising blood serum. Serum levels of retinol and ${\alpha}$-tocopherol among cases ($0.177{\pm}0.081$, $1.649{\pm}1.670{\mu}g/ml$) were significantly lower than in controls ($0.264{\pm}0.137$, $3.225{\pm}2.054{\mu}g/ml$) (p<0.005). Although serum level of ${\beta}$-carotene among cases ($0.106{\pm}0.159{\mu}g/ml$) were lower compared to controls ($0.134{\pm}0.131{\mu}g/ml$), statistical significance was not observed. Logistic regression analysis showed that high serum level of retinol (OR=0.501, 95% CI=0.254-0.992, p<0.05) and ${\alpha}$-tocopherol (OR=0.184, 95% CI=0.091-0.370, p<0.05) was significantly related to lower risk of oral cancer, whereas no relationship was observed between ${\beta}$-carotene and oral cancer risk. Conclusions: High serum levels of retinol and ${\alpha}$-tocopherol confer protection against oral cancer risk.

• 한국환경복원기술학회지
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• 제18권6호
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• pp.151-165
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• 2015

This study surveys and analyzes the ecology of Gangseo area to be developed near Nakdonggang River and West Nakdonggang River in Busan - before its development. With the result of the surveys as its basis, the study also finds ways to conserve the area's bird habitats in response to the planned development. According to the site surveys over four seasons, West Nakdonggang River which is near the targeted development area is a wintering ground for migratory birds. Some representative bird types in this area are ducks, geese, and other water birds. There are even six legally protected species. The average altitude of their flight is calculated to be $92.4{\pm}18.8m$ and the main purpose for the migration is determined to be for food. According to evalution of the conservation value of the area, Nakdonggang River and West Nakdonggang River turns out to have the highest conservation value. Other areas are developable as long as it was environmentally friendly. Therefore, this study suggests incorporation of ecological parks and biotopes within the targeted development area along with the proposal for spaces in the surroundings where the wintering birds can rest and feed. Especially for the wintering birds that rest at Nakdonggang River and West Nakdonggang River, their flight direction and pattern was examined to determine that their resting ground should be located 50~100m off of the river bank. Furthermore, this study proposes ways to maximize bird habitat by building ecological corridors between the agricultural channel and other small streams in the targeted development area and the habitats in the surroundings. Lastly, the study suggests the construction of low buildings, at most 20m in height, near Nakdonggang River and West Nakdonggang River considering the birds' flight altitude and direction.

• BMB Reports
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• 제40권4호
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• pp.459-466
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• 2007

The cDNA sequence of the Japanese flounder (Paralychthys olivaceus) IgD has been previously reported (GenBank accession no. AB052658) and this was followed by the detection of IgD mRNA expression in some flounder organ tissues. However, it has not been determined whether the flounder IgD gene is virtually expressed into IgD protein. To characterize the flounder immunoglobulins utilized in elucidating the mechanism, evolution and diversity of the flounder immune system, antibodies specific to IgD and IgM were necessary. In the present study, partial flounder recombinant IgD (rIgD), IgM (rIgM) and the conserved regions of IgD and IgM (rCIg) were produced by cloning the cDNA sequence using isotype specific primers which were designed to produce unique fragments of IgD and IgM specific amino acid sequences. The production of recombinant Igs was ascertained by SDS-gel electrophoresis and immunoblot analysis using anti-T7$\cdot}$Taq antibody. The produced recombinant Igs were purified using affinity columns, and used as immunogens. Antibodies specific to the isotype of flounder Igs were generated by immunizing rabbits with rfIgs and the antibodies produced were identified by enzyme-linked immunosorbent assay (ELISA) and immunoblotting. Specificities of the generated antibodies were evaluated by testing cross-reactivity between recombinant IgM and IgD. By ELISA, rabbit antibodies against the rfIgD fragment (anti-rfIgD) failed to recognize any kind of flounder serum Igs, whereas respective antibodies against rfCIg (anti-rfCIg) and rfIgM fragments (anti-rfIgM) reacted with serum Igs. Likewise, in immunoblot assays, though anti-rfIgD did not, both anti-rfCIg and anti-rfIgM bound with the ~85 kd flounder IgM heavy chain. By flow cytometry analysis, anti-rfCIg, anti-rfIgD and anti-rfIgM reacted with 6%, 3% and 6.5% of cells, respectively, suggesting that flounder IgD is not secreted in serum but expressed on flounder B-like cell surfaces as in mammals. Antibodies produced against recombinant flounder Igs could be used to develop sandwich assay systems for detecting flounder Igs and for further investigating the flounder immune system.

• 한국환경농학회지
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• 제32권2호
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• pp.155-165
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• 2013

본 연구에서 문헌 조사를 통하여 우리나라에서 논을 이용하는 조류의 현황을 조사하였다. 논을 이용하는 조류는 논의 본답, 논둑, 저수지 또는 관개 수로에서 번식, 휴식 또는 취식을 하는 조류로 정의하였다. 1980년대 이후부터 출판된 자료에 따르면, 논을 이용하는 조류는 총 47과 279종이었다. 도요 물떼새과(18%), 매, 수리 및 올빼미과의 맹금류(12%), 오리과 (11%)의 조류가 논을 주로 이용하였다. 논 이용 조류 중 절반은 겨울철에 건답을 이용하였으며, 39.4%는 봄이나 가을 또는 여름에 습답을 이용하는 종이었다. 논 의존도는 오리류, 백로류 및 도요류가 높았다. 수면성오리 개체군은 지난 12년 동안 감소한 반면에 두루미는 증가하는 경향을 보였다. 높은 논 의존도를 갖는 멸종위기종은 두루미, 재두루미, 검은목두루미, 흑두루미, 개리, 흰이마기러기와 따오기였다. 논은 다수의 멸종위기종을 포함하여 다양한 조류에게 사계절 내내 중요한 서식지로 제공되고 있다.

• 한국환경생태학회지
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• 제24권3호
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• pp.318-324
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• 2010

이 연구의 목적은 고라니(Hydropotes inermis argyropus)의 위내용물을 대상으로 PCR-DGGE 방법을 이용하여 그 식이습성을 조사하는데 있다. 이 연구를 위해, 강원도 철원과 전라남도 동부지역 등에서 자연사 혹은 로드킬에 의해 죽은 고라니 사체의 위에서 식이물 샘플을 채취하였다. 총 44개체의 위내용물에서 각각 DNA를 추출하였고, 두 가지의 프라이머(rbcLZ1과 rbcL19bR)를 이용하여 ribulose-1,5-bisphosphate carboxylase large subunit(rbcL) gene을 PCR 증폭하였다. 44개의 샘플 중 29 샘플에서 성공적으로 PCR을 수행하였다. 이 29개 partial rbcL gene의 PCR product는 PCR-DGGE에 이용되었다. 식이물에 대한 분석결과 총 6과의 식물이 확인되었다. 강원도 철원의 경우, 5과가 나타난 반면, 전라남도 동부의 경우, 3과만이 확인되었다. 이 연구에서는 종수준의 먹이식물의 구별에는 실패하였 지만, 차후 이 PCR-DGGE 기법은 고라니를 포함한 초식동물의 식이습성을 분석하는데 하나의 가능성 있는 방법이 될 것으로 생각된다.

• 한국작물학회:학술대회논문집
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• 한국작물학회 2017년도 9th Asian Crop Science Association conference
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• pp.104-104
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• 2017

Sorghum (Sorghum bicolor (L.) Moench.) is one of the most important crops for human and animal nutrition. Nonetheless, sorghum has a cyanogenic glucoside compound which can be degraded into hydrogen cyanide, toxic to humans and animals even with tiny amount. In consequence, breeding materials with a low cyanide level has been a top priority in sorghum breeding programs. To fulfill our long-term goal, we are screening sorghum accessions with low cyanide level, which would be an important breeding material for food safety. We collected seeds of various sorghum accessions and analyzed relevant metabolites to find useful breeding materials of sorghum accessions containing low cyanide. Fourteen wild relatives were obtained from the University of Georgia in US, a reference accession BTx623, and three local varieties from National Agrobiodiversity Center of Rural Development Administration in Korea, and one wild species from the Wild Plant Resources Seed Bank of Korea University in Korea. Sorghum plants were grown in plastic greenhouse under natural conditions. After growing, leaf samples were harvested at different developmental stages: seedling phase, vegetative phase (right before flowering), and reproductive phase (ripening). Using collected samples, quantification analysis were performed by an HPLC system for three metabolites (dhurrin, 4-hydroxybenzaldehyde, and 4-hydroxyphenylacetic acid) in sorghum plants. Prior to metabolome analysis, specific experimental condition for HPLC system was set to be able to separate three metabolites simultaneously. Under this condition, these metabolites were quantified in each accession by HPLC system. We observed that the metabolite contents were changed differently by developmental stages and accessions. We clustered these results into five groups as patterns of their contents by developmental stages. Most of accessions showed that 4-hydroxybenzaldehyde content was very high at seedling stage and decreased rapidly at vegetative phase. Interestingly, the patterns of dhurrin content were very different among clusters. However, 4-hydroxyphenylacetic acid content was maintained at low levels by developmental stages in most accessions. The results would demonstrate how dhurrin and alternative degradation pathways are differentiated in each accession.

• 대한지역사회영양학회지
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• 제13권4호
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• pp.582-591
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• 2008

Both genetic and environmental factors play important roles in the pathogenesis of Parkinson Disease(PD). The contribution of many environmental factors including dietary factor remains unproven. The purpose the study was to investigate the dietary habits, nutrient intake and dietary quality of Korean PD patients according to the duration of disease. PD patients were recruited from K and S university hospitals from May 2005 to January 2006. This study was carried out after approval by the Institute Review Board(IRB). British Brain Bank criteria was used to diagnose PD. The subjects were classified into 2 groups based on the duration of PD: <25 months and $\geq$25 months groups. General characteristics, anthropometric measurements, food habits and dietary intakes were investigated. The results of this study were as follows: 1) The mean age of <25 months group($66.9{\pm}8.0$ yr) was significantly higher than that of $\geq$25 months group($62.2{\pm}8.8$ yr)(p<0.05). No significant differences were found for academic background, occupation, living status and social activity, however, numbers of diseases, exercise and family history of PD were significantly different. 2) Anthropometric measurements were not different between the two groups. 3) The frequency of taking snacks was significantly higher in <25 months group and the amounts of alcohol consumption were significantly higher in $\geq$25 months group. 4) Daily intakes of most nutrients were very low compared with DRI. 5) The MAR score was significantly lower in <25 months group(p<0.05;) however, the scores of DVS, DDS and DQI were not significantly different. As a conclusion an overall nutrient intake and dietary quality of the Parkinson's Disease patients need to be improved regardless of duration of the disease and a well-balanced diet should be emphasized.

• 한국균학회지
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• 제42권4호
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• pp.282-288
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• 2014

본 연구에서는 순창지역에서 만들어지는 장류에서 곰팡이를 분리하고 동정하여 보다 안전하고 기능성이 높은 발효제품을 위한 균주를 확보하고자 하였다. 순창지역에서 생산되는 장류 제품으로부터 곰팡이를 분리하여 ${\beta}$-tubulin 유전자 분석 통해 10개의 균주가 Aspergillus oryzae/flavus complex임을 알 수 있었다. 보다 정확한 동정을 위하여 아플라톡신 클러스터 유전자 중에 하나인 omtA의 염기서열을 증폭하여 A. oryzae와 A. flavus 표준 균주의 omtA 서열과 함께 계통 분류한 결과, A. oryzae의 표준 균주와의 유연관계가 높음을 알 수 있었다. 또한 norB-cypA 사이의 염기서열을 증폭한 결과 500 bp이 증폭 산물이 확인되었는데 이는 표준 균주인 A. oryzae의 norB-cypA 사이의 염기서열 증폭 산물과 동일한 크기임을 확인할 수 있었다. A. oryzae로 확인된 10균주를 활용하여 코지를 제조하고 ${\alpha}$-amylase 활성과 protease 활성을 측정하였다. Protease 활성은 6, 13, 17, 27, 37, 그리고 38 균주로 제조된 코지는 대조구(시판되고 있는 종균으로 제작한 코지)보다 2배 정도 높은 protease 활성을 보였으며, ${\alpha}$-amylase 활성은 257~320 U/mL로 측정되었다. 식품안전성을 위한 아플라톡신 분비 확인 결과, 63번 균주로 제조된 코지를 제외한 모든 코지에서 아플라톡신을 만들지 않는 것으로 확인되어, 순창에서 분리된 A. oryzae는 추후 메주 접종균으로 개발할 수 있음을 보여주었다.

• 한국수자원학회:학술대회논문집
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• 한국수자원학회 2019년도 학술발표회
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• pp.176-176
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• 2019

Thailand had issued a national strategic development master plan with issues related to water resources and water security in the entire water management. Water resources are an important factor of living and development of the country's socio-economy to be stable, prosperous and sustainable. Therefore, water management in both multidimensional and multi-sectoral systems is important and will supports socio-economic and environmental development. The direction of national development in accordance with the national strategic framework for 20 years that requires the country to level up security level in terms of water, energy and food. To response to the proposed goals, there is a subplan to increase water productivity of the entire water system for economical development use by evaluating use value and to create more value added from water use to meet international standard level. This study aims to evaluate the water productivity of Thailand in each basin and all sectors such as agricultural sector, service and industrial sectors by using the water use data from water account analysis and GDP data from NESDB during the past 10 years (1996-2015). The comparison of water productivity with other countries will also be conducted and in addition, the measures to improve water productivity in next 20 years will be explored to response to the National Strategic Master Plan goals. Water productivity is defined as output per unit of water depleted. The simplest way to compare water productivity across different enterprises is in monetary terms. World Bank presents water productivity as an indication of the efficiency by which each country uses its water resources. There are two data sets used for water productivity analyses, i.e., the first is water use data at end users and the second is Gross Domestic Product. The water use at end users are estimated by water account method based on the System of Environmental-Economic Accounting for Water (SEEA-Water) concept of United Nations. The water account shows the analyses of the water balance between the use and supply of each water resource in physical terms. The water supply and use linkage in the water account analyses separated into each phases, i.e., water sources, water managers, water service providers, water user at end user under water regulators of all kinds of water use activities such as household, industrial, agricultural, tourism, hydropower, and ecological conservation uses. The Gross Domestic Product (GDP), a well- known measuring method of the national economic growth is not actually a comprehensive approach to describe all aspects of national economic status, since GDP does not take into account the costs of the negative impacts to natural resources that result from the overexploitation of development projects, however, at present, integrating the environment with the economy of a country to measure its economic growth with GDP is acceptable worldwide. The study results will show the water use at each basin, use types at end users, water productivity in each sector from 1996-2015 compared with other countries, Besides the productivity improvement measures will be explored and proposed for the National Strategic Master Plan.