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Evolutionary Explanation for Beauveria bassiana Being a Potent Biological Control Agent Against Agricultural Pests

  • Han, Jae-Gu
    • 한국균학회소식:학술대회논문집
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    • 2014.05a
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    • pp.27-28
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    • 2014
  • Beauveria bassiana (Cordycipitaceae, Hypocreales, Ascomycota) is an anamorphic fungus having a potential to be used as a biological control agent because it parasitizes a wide range of arthropod hosts including termites, aphids, beetles and many other insects. A number of bioactive secondary metabolites (SMs) have been isolated from B. bassiana and functionally verified. Among them, beauvericin and bassianolide are cyclic depsipeptides with antibiotic and insecticidal effects belonging to the enniatin family. Non-ribosomal peptide synthetases (NRPSs) play a crucial role in the synthesis of these secondary metabolites. NRPSs are modularly organized multienzyme complexes in which each module is responsible for the elongation of proteinogenic and non-protein amino acids, as well as carboxyl and hydroxyacids. A minimum of three domains are necessary for one NRPS elongation module: an adenylation (A) domain for substrate recognition and activation; a tholation (T) domain that tethers the growing peptide chain and the incoming aminoacyl unit; and a condensation (C) domain to catalyze peptide bond formation. Some of the optional domains include epimerization (E), heterocyclization (Cy) and oxidation (Ox) domains, which may modify the enzyme-bound precursors or intermediates. In the present study, we analyzed genomes of B. bassiana and its allied species in Hypocreales to verify the distribution of NRPS-encoding genes involving biosynthesis of beauvericin and bassianolide, and to unveil the evolutionary processes of the gene clusters. Initially, we retrieved completely or partially assembled genomic sequences of fungal species belonging to Hypocreales from public databases. SM biosynthesizing genes were predicted from the selected genomes using antiSMASH program. Adenylation (A) domains were extracted from the predicted NRPS, NRPS-like and NRPS-PKS hybrid genes, and used them to construct a phylogenetic tree. Based on the preliminary results of SM biosynthetic gene prediction in B. bassiana, we analyzed the conserved gene orders of beauvericin and bassianolide biosynthetic gene clusters among the hypocrealean fungi. Reciprocal best blast hit (RBH) approach was performed to identify the regions orthologous to the biosynthetic gene cluster in the selected fungal genomes. A clear recombination pattern was recognized in the inferred A-domain tree in which A-domains in the 1st and 2nd modules of beauvericin and bassianolide synthetases were grouped in CYCLO and EAS clades, respectively, suggesting that two modules of each synthetase have evolved independently. In addition, inferred topologies were congruent with the species phylogeny of Cordycipitaceae, indicating that the gene fusion event have occurred before the species divergence. Beauvericin and bassianolide synthetases turned out to possess identical domain organization as C-A-T-C-A-NM-T-T-C. We also predicted precursors of beauvericin and bassianolide synthetases based on the extracted signature residues in A-domain core motifs. The result showed that the A-domains in the 1st module of both synthetases select D-2-hydroxyisovalerate (D-Hiv), while A-domains in the 2nd modules specifically activate L-phenylalanine (Phe) in beauvericin synthetase and leucine (Leu) in bassianolide synthetase. antiSMASH ver. 2.0 predicted 15 genes in the beauvericin biosynthetic gene cluster of the B. bassiana genome dispersed across a total length of approximately 50kb. The beauvericin biosynthetic gene cluster contains beauvericin synthetase as well as kivr gene encoding NADPH-dependent ketoisovalerate reductase which is necessary to convert 2-ketoisovalarate to D-Hiv and a gene encoding a putative Gal4-like transcriptional regulator. Our syntenic comparison showed that species in Cordycipitaceae have almost conserved beauvericin biosynthetic gene cluster although the gene order and direction were sometimes variable. It is intriguing that there is no region orthologous to beauvericin synthetase gene in Cordyceps militaris genome. It is likely that beauvericin synthetase was present in common ancestor of Cordycipitaceae but selective gene loss has occurred in several species including C. militaris. Putative bassianolide biosynthetic gene cluster consisted of 16 genes including bassianolide synthetase, cytochrome P450 monooxygenase, and putative Gal4-like transcriptional regulator genes. Our synteny analysis found that only B. bassiana possessed a bassianolide synthetase gene among the studied fungi. This result is consistent with the groupings in A-domain tree in which bassianolide synthetase gene found in B. bassiana was not grouped with NRPS genes predicted in other species. We hypothesized that bassianolide biosynthesizing cluster genes in B. bassiana are possibly acquired by horizontal gene transfer (HGT) from distantly related fungi. The present study showed that B. bassiana is the only species capable of producing both beauvericin and bassianolide. This property led to B. bassiana infect multiple hosts and to be a potential biological control agent against agricultural pests.

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Chemical Synthesis and Orientation Disruption Bioassay of Sex Pheromone of the Oriental Fruit Moth, Grapholita molests (Busck) (복숭아순나방(Grapholita molests) 성페로몬 합성과 유인력 교란생물검정)

  • Kim, Yong-Gyun;Bae, Sung-Woo;Bae, Soo-Il;Yoon, Hyang-Mi;Hong, Yong-Pyo
    • Korean journal of applied entomology
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    • v.45 no.3 s.144
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    • pp.309-316
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    • 2006
  • Three sex pheromone components (cis-8-dodecenyl acetate (Z12Ac), trans-8-dodecenyl acetate (E12Ac), cis-8-dodecenol (Z12OH)) of the Oriental fruit moth, Grapholita molesta, were chemically synthesized. Especially to increase the composition of cis-stereoisomer, a triple bond intermediate was hydrogenated at $-20^{\circ}C$ with catalytic $Pd/BaSO_{4}$. The resulting product consisted of the acetates with a stereoisomer ratio in 92:8 (Z:E). The biological activity of the synthesized pheromone compounds was analyzed both in male responses and orientation disruption. The indoor pheromone effect was determined by male flight behavior showing wing movement in response to lure. Different mixtures of the synthetic pheromone components were prepared by mixing acetate and alcohol components in 100:0, 99:1, and 90:10 (g/g) and tested with a comparison of a standard commercial pheromone lure. The highest pheromone effect was observed in only acetate mixture (100:0) and the effect was reduced with the addition of the alcohol component. This indoor pheromone effect could be observed in field monitoring trial, in which 100:0 mixture showed the highest trap catches. Orientation disruption assay was conducted indoor by using a cage, in which the center had a commercial lure on sticky plate and the four candidates were placed at 6 cm away from the central lure on each of four directions. Test males were released to the arena during overnight (12 h) and then the caught males on the sticky plate were counted. The synthesized pheromone as well as the commercial pheromone showed 100% orientation disruption. However, the orientation disruption effect was reduced with decrease in the number of the surrounding disrupting pheromone baits. These results clearly suggest that the synthesized sex pheromone of G. molesta is biologically active and can be used for field mating disruption.

The actual conditions on drug abuse among High school students in Busan city (고등학생(高等學生)의 약물(藥物) 남용(濫用) 실태(實態))

  • Cho, Yeon-Sook
    • Journal of the Korean Society of School Health
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    • v.3 no.1
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    • pp.101-118
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    • 1990
  • This study was intended to investigate actual conditions and problems relating to a variety of substance which have been used by students. A Questionnaire survey was carried out with the subject of 2411 students in an academic boys' high school, academic girls' high school, technical boys' high school and technical girls' high school in Pusan, from the 15th day to 29th day of March, 1989. The summarized results were as follows. There was not a remarkable difference in distribution of these subjects in boys' & girls' high school. The common and good health condition of subjects accounted for 90 percent or higher. 24.4 percent of them also had smoking experience, 11.6 percent of which continues to smoking. The understanding rate of these substances name other than sedative, psychostimulants and antihypnotic accounted for 90 percent or higher. The experience rate taking these substances for one year showed that anodyne, digestive and nutritive tonic accounted for 70 percent, antihypnotic for 15.6 percent, sedative for 1.4 percent, respectively, and psychostimulants for 0.5 percent. Moreover. it was shown that drugs accounted for 1.5 percent, bond for 1.4 percent, and thinner for 0.5 percent. The rate of the daily experiencers who took anodyne, digestive, nutritive tonic, sedatives, and psychostimulants and so on was 7.7 percent, 6.2 percent, 5.2 percent, 5.9 percent, and 5.0 percent respectively. This fact implied that there was a serious problem in high school students' non - narcotic abuse. The usage rate of these substances for treatment purpose showed that anodyme accounted for 90 percent or higher, and digestive for 70-80 percent, respectively, where girl students showed higher rate than one of boy students. Additionally, there was higher the usage rate for other purposes. The usage rate of drugs was highest when these students felt melancholy and curiousity. Their obtaining place appeared that these students mainly obtained these drugs, bonds and thinners from a small shop or peddler and their friend while they usually obtained other substance from the pharmacy and medical institute. The first usage time of these substances appeared during the middle school (the age of 14-15) which was the highest rate. The smoker of all subjects used remarkly large substances as compared with one of no-smoker. Particularly, it appeared that the usage of drugs was very closely related to smoking. The large number of students did not use these substances for oesrable purposes even though they understood the name of these substances. For this reason, from primasy schools it is required to teach the students dependence and harmful effects caused by these substances abuse. Moreover, it was shown that these students firstly used these substances during the middle school (the age of 14-15)due to curiosity. As a result, it is very urgent to give the students health education suitabale for prevention of these substance abuse, when considering harmful effects of these substances. And so health education for no-smoking. Finally, considering that it is very easy to obtain these substances from a small shop and pharmacy, the regulation of these substances control should be considered and completed in the future.

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Comparison of Direct-labeling Method of Antibody with $^{99m}Tc$ and $^{188}Re$ (농양이식백서에서 $^{99m}Tc,\;^{188}Re$ 직접표지항체의 비교)

  • Choi, Tae-Hyun;Lim, Sang-Moo;Choi, Chang-Woon;Woo, Kwang-Sun;Chung, Wee-Sup;Lim, Soo-Jeong
    • The Korean Journal of Nuclear Medicine
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    • v.33 no.1
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    • pp.84-93
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    • 1999
  • Purpose: We investigated the direct labeling method of antibody with $^{99m}Tc$ and $^{188}Re$ and examined the stability and function of these labeled compounds in in vitro and in vivo. Materials and Methods: Disulfide bond of nonspecific human IgG was reduced to -SH group by 2-mercaptoethanol. Stannous ion was used to reduce $^{99m}Tc$ and $^{188}Re$. The stability of $^{99m}Tc$-IgG and $^{188}Re$-IgG was estimated upto 24 hrs. Biodistribution was evaluated in abscess bearing rats at 4 and 24 hr post-injection of $^{99m}Tc$ or $^{188}Re$ labeled IgG. Results: The number of -SH group per reduced IgG molecule was 2.34. The labeling yield of $^{99m}Tc$-IgG and $^{188}Re$-IgG were 90% and 95%, respectively The stability of $^{99m}Tc$-IgG at 1, 4, 6 and 24 hr was 91%, 83%, 78%, 7% and that of $^{188}Re$-IgG at 1, 4, 16 and 24 hr was 94%, 80%, 47%, 42%, respectively. At 4 hr post-injection of $^{99m}Tc$-IgG, high uptake was found on kidney, blood, stomach and abscess ($9.42{\pm}0.68,\;1.43{\pm}0.24,\;0.86{\pm}0.18,\;0.72{\pm}0.10$ %ID/g, respectively). The uptakes at 24 hr were kidney, abscess,.itomach, and blood in descending order. In case of $^{188}Re$-lgG, high uptake at 4 hr post injection appeared on kidney, blood, abscess and stomach ($3.92{\pm}0.62,\;1.32{\pm}0.08,\;0.88{\pm}0.01,\;0.26{\pm}0.06$, respectively). The uptakes at 24 hr were kidney, abscess, blood and stomach in descending order. The abscess to blood uptake ratio of $^{99m}Tc$-IgG was 0.5 at 4 hr and 2.02 at 24 hr and that of $^{188}Re$-IgG was 0.67 and 1.29. Conclusion: $^{99m}Tc$-IgG and $^{188}Re$-IgG canbe labeled efficiently with direct labeling method. However, $^{99m}Tc$-IgG and $^{188}Re$-IgG, labeled with direct method, was unstable. Further study is needed to enhance the stability of the antibody labeling.

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Tensile Behavior of Hybrid Fiber Reinforced Cement Composite According to the Hooked Steel Fiber and Polyvinyl Alcohol Fiber Blending Ratio and Strain Rate (후크형 강섬유와 폴리비닐알코올 섬유의 혼합 비율과 변형속도에 따른 하이브리드 섬유보강 시멘트복합체의 인장거동)

  • Son, Min-Jae;Kim, Gyu-Yong;Lee, Sang-Kyu;Kim, Hong-Seop;Nam, Jeong-Soo
    • Journal of the Korea institute for structural maintenance and inspection
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    • v.21 no.6
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    • pp.98-105
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    • 2017
  • In this study, the fiber blending ratio and strain rate effect on the tensile behavior of hybrid fiber reinforced cement composite was evaluated. Hooked steel fiber and polyvinyl alcohol fiber were used for reinforcing fiber. The fiber blending ratio of HSF+PVA were 1.5+0.5, 1.0+1.0 and 0.5+1.5vol.%. As a results, the tensile strength, strain capacity and fracture toughness of the hooked steel fiber reinforced cement composites were improved by the increase of the bond strength of the fiber and the matrix according to increase of strain rate. However, the tensile stress sharply decreased after the peak stress because of the decrease in the number of straightened pull-out fibers by micro cracks in the matrix around hooked steel fiber. On the other hand, PVA fiber showed cut-off fracture at strain rate $10^{-6}/s$ with multiple cracks. However, at the strain rate $10^1/s$, the multiple cracks and strain capacity were decreased because of the pull-out fracture of PVA fiber. The HSF1.5PVA0.5 shown the highest tensile strength because the PVA fiber suppressed the micro cracks in the matrix around the hooked steel fiber and improved the pull-out resistance of hooked steel fiber. Thus, DIF of strain capacity and fracture toughness of HSF1.5PVA were greatly improved. In addition, the synergistic response of fracture toughness was positive because the tensile stress was slowly decreased after the peak stress by improvement of the pull-out resistance of hooked steel fiber at strain rate $10^1/s$.

Studies on the Effects of Copper on the Lactate Dehydrogenase and Esterase Isozymes in Various Tissues of Carassius carassius (붕어(Carassius carassius)의 조직내 젖산수소이탈효소와 에스테라아제 아이소자임에 미치는 동의 영향에 관한 연구)

  • Lee, Choon-Koo;Choo, Il-Young
    • The Korean Journal of Zoology
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    • v.16 no.2
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    • pp.79-96
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    • 1973
  • In order to elucidate the effects of copper on Corassius carassius, the following were studied: 1) lactate dehydrogenase isozyme patterns by cellulose acetate electrophoresis, 2) LDH activity and copper effect on LDH enzyme system y spectrophotometry, 3) esterase isozyme patterns by agar thin layer electrophoresis, 4) hemoglobin patterns by starch gel electrophoresis, and 5) histological study. 1. There were two bands of LDH isozymes (LDH-3 and LDH-5) in the gill, three bands (LDH-2, LDH-4, and LDH-5) in the liver, and two bands (LDH-3 and LDH-4) in the muscle of the normal fish. The LDH-1 bond was not found in the above three tissues. When the fish were exposed to copper, LDH-3 appeared in the liver, LDH-5 in the muscle, but no new LDH band appeared in the gill. 2. The sepcific activities of the LDH were lowest in the gill and highest in the muscle of the normal fish, and they were gradually decreassed in the gill and highest in the muscle of the normal fish, and they were gradually decreased in the liver and mucle except in the gill from 1-day to 10-day exposure to copper. It indicates that LDH activities in the liver and muscle of the fish were inhibited by copper. 3. Through in vitro experiment, it is clear that the decrease of the LDH activities of the liver and muscle of the fish exposed to copper is mainly caused by the inhibition on the M-LDH in the fish. 4. The numbers of the esterase isozyme bands of the gill, liver, muscle, blood, brain, and kidney of the normal fish were 3, 6, 2, 2, 2, and 2 respectively, and these numbers were the same as those exposed to copper. The relative mobilities of the esterase bands in the gill, liver, blood, and kidney of the exposed group were different from those of the control. 5. There was one hemoglobin band on the anode in the normal fish. It seems that the nobility of hemoglobin band of the fish exposed to copper was slightly faster than that of the normal fish. 6. The normal gill lamellae of the fish consisted of centrally located pillar cells and a number of mucus cells. When the fish were exposed to copper, the epithelial layer was divorced first, disintegrated, and then destroyed completely. 7. The liver of the normal fish had prominent central veins, cords of hepatic cells, and sinusoids. When the fish were exposed to copper, numerous droplets of fat appeared in the cells around the central vein of the liver. It is assumed that the fatty droplets were accumulated by the lesion due to fatty metamorphosis of the liver caused by copper. 8. There was no histological difference between the muscle of the normal fish and that of the fish exposed to copper. 9. In the normal fish, the tubules of the kidney were surrounded by hemopoetic tissues. However, the kidney tissue of the fish exposed to copper received some damage on the proximal tubules. Since the tubule cells were reduced in height, the lumens of the tubules were enlarged. Consequently many proximal tubules exhibited some pink-stained granular casts and various stages of degeneration.

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An Exploratory Study on Social Presence in Synchronous Distance Course : Focused on the Cases of Christian Education Classes (실시간 화상 수업에서의 사회적 실재감 탐색 : 기독교교육 수업 사례를 중심으로)

  • Park, Eunhye;Sung, Jihoon
    • Journal of Christian Education in Korea
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    • v.64
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    • pp.203-235
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    • 2020
  • The purpose of this study is to identify the degree of social presence perceived by students and to explore the factors that have affected it after practicing Christian Education classes as synchronous distance course due to Covid-19. It is also to suggest effective ways in the aspects of the design and operation to improve social presence. In order to measure social presence and derive influencing factors, research related to synchronous distance class and social presence is summarized through literature review. The researchers also surveyed 58 students in three courses of Christian education major at a University in Gyeonggi-do and conducted in-depth interviews with 6 students. The main findings are as follows: First, the sense of social presence was moderate, the emotional bond was the lowest by sub-factor, the open communication, the sense of community was moderate, and the mutual support and concentration were the highest. Second, factors that had a positive impact on the sense of social reality were group activities, selfintroduction activities, active participation in classes, mutual communication such as Q & A or response to peer learners' opinions during lectures by professors, questions, feedback, etc, and having a smaller number of students. Factors that had a negative impact on the perception of social presence were lack of private conversations, poor participation in classes, lack of communication with each other, and difficulty concentrating. The causes of these negative factors were technical problems and limitations arising from zoom, inconvenience and distracting surroundings, lack of time, and psychological awkwardness. Reflecting the results of the study, orientation to effective synchronous distance course, guidance on smooth communication methods, strengthening the role of professors to promote learning, strengthening group activities and learner-centered activities, and proposing a smaller scale of students were ways that are offered to improve the sense of social presence in synchronous distance courses.

Study on the Factors Influencing the Investment Performance of Domestic Venture Capital Funds (국내 벤처펀드의 투자성과에 영향을 미치는 요인에 관한 연구)

  • InMo Yeo;HyeonJu Park;KwangYong Gim
    • Asia-Pacific Journal of Business Venturing and Entrepreneurship
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    • v.18 no.5
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    • pp.63-75
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    • 2023
  • This study conducted empirical analysis on the factors affecting the investment performance of 205 domestic venture funds (with a total liquidation amount of 7.25 trillion KRW) newly formed from 2007 to 2017 and completely liquidated as of the end of 2022. Due to the nature of private equity funds, obtaining empirical data is extremely challenging, especially for data post-COVID-19 era liquidations. Nevertheless, despite these challenges, it is meaningful to analyze the impact on the investment returns of domestic venture funds using the most recent data available from the past 10 years. This study categorized the factors influencing venture fund performance into external environmental factors and internal factors. External environmental factors included "economic cycles," "stock markets," "venture markets," and "exit markets," while internal factors included the fund management company's capabilities in terms of "experience," "professional personnel," and "assets under management (AUM)." The fund structure was also categorized into "fund size" and "fund length" for comparative analysis. In summary, the analysis yielded the following results: First, the 3-year government bond yield, which represents economic cycles well, was found to have a significant impact on fund performance. Second, the average 3-month KOSDAQ index return after fund formation had a statistically significant positive effect on fund performance. Third, the number of IPOs, indicating the competition intensity at the time of venture fund liquidation, was shown to have a negative effect on fund performance. Fourth, it was observed that the larger the AUM of the fund management company, the better the fund's returns. Finally, venture fund returns showed variations depending on the year of formation (Vintage). Therefore, when individuals consider investing in venture funds, it is considered a highly effective investment strategy to construct an investment portfolio taking into account not only external environmental factors and internal fund factors but also the vintage year.

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Cultural Education Methods for Overseas Koreans Using Classical Narratives: Focusing on Princess Bari and The Tale of Shim Cheong (고전 서사무가를 활용한 재외동포의 문화 교육 방안 연구 - <바리공주>와 <심청전>을 중심으로 -)

  • Kang Myung-ju
    • Journal of the Daesoon Academy of Sciences
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    • v.47
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    • pp.173-202
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    • 2023
  • In this study, we delve into the potential for innovative cultural education techniques that utilize the timeless tales of Princess Bari and The Tale of Shim Cheong as tailored for the upcoming generations of overseas Korean learners. With a rising number of young overseas Koreans born and raised in their host countries, there emerges a pressing need to craft an educational framework that resonates with the evolving dynamics of their generation. Our endeavor revolves around proposing educational strategies that help solidify identity while carefully considering the intrinsic motivation prevalent among most overseas Koreans. Naturally, the choice of employing the classic epics Princess Bari and The Tale of Shim Cheong as educational resources was deliberate. These narratives are rich in rites of passage and offer profound insights into the transformative journey of their protagonists. Both characters are affluent women in patriarchal societies, and both embark on quests to redefine themselves through new relationships, liberating themselves from the confines of parental ties. This narrative framework provides a unique opportunity for overseas Koreans who are often adrift in the social fabric of their adopted countries. These stories inspire them to introspect and contemplate their own identities. By intertwining their personal narratives with the empowering stories of characters, students are provided a chance to reaffirm their authentic selves. Therein, a paradigm shift can occur that allows individuals to embrace the core elements that define them. Our ultimate objective was to enable students to explore their own stories and immerse themselves in the intricate narratives of classical works. This immersive experience fosters a profound sense of unity with the characters and paves the way for a comprehensive educational plan. This plan not only celebrates the hybrid nature of identity but also cultivates a deep sense of positivity within amalgamated 'subjects.' Such an approach not only fosters a stronger connection with one's heritage but also sparks a genuine curiosity about and affinity for the rich cultural tapestry of one's home country. It's not just education; it's a transformative journey that enriches the lives of overseas Koreans and nurtures a profound bond with their cultural roots.

A Study on Health Awareness of Middle and High School Students in Yong Nam Area (영남지역(嶺南地域) 중고등학교학생(中高等學校學生)들의 보건의식행태조사(保健意識行態調査) 연구(硏究))

  • Kim, Hyung Nam;Nam, Chul Hyun
    • Journal of the Korean Society of School Health
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    • v.4 no.2
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    • pp.119-135
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    • 1991
  • The study was designed to gain necessary basic data order to grasp health knowledge, attitude, practice level of middle and high school students and to analyse th problem and to point out the method of improvement in the field of school health education. The survery was carried out through this reporter's interview for 2,400 students who attend to ten schools in Young Nam area during the period of a month from 25 the June to 25th July 1989. The result of this study can be summaried as follows. 1. The total number of answers on the question was 2,346. As for general characteristics the percent of female middle school students was 60.6% and the percent of male students was 77.7%, 45.9% of high school students was evening school students. 52.9% of middle school students and 42.3% of high school students were borne in rural area. 2. The percentage of unknown and misunderstanding for Epidemic Hepatitis infection was 46.3% of middle school students and 29.6% of high school students. 3. The percentage of unknown and misunderstanding for Epidemic Hemorrhage fever infection was 85.6% of middle school students and 66.9% of high school students. 4. The percentage of right knowledge for AIDS infection was 66.0% of middle school students and 90.4% of high school students. 5. The percentage of right knowledge for Typhoid infection was 47.8% of middle school students and 69.4% of high school students. 6. The percentage of unknown and misunderstanding for Tuberculosis infection was 71.6% of middle school students and 62.2% of high school students. 7. As for personal hygiene, the percentage of toothbrushing after every meal was high level : 44.2% of middle school students and 42.0% of high school students. 8. 60.9% of middle school students take a bath twice a week, 49.2% oh high school students take a bath a week. Times of bath of middle school students was higher than that of high school students. 9.The percentage of washing hand after using toilet was 42.1% of middle school students and 35.1% of high school students. 49.0% of middle school students and 55.1% of high school students wash hand sometimes after using toilet. 10. The percentage of change of underwear twice a week was 57.6% of middle school students and 49.8% of high school students. 11. The percentage of habit of unbalanced diet was 30.% of middle school students and 27.6% of high school students. 50.8% of middle school students and 51.7% of high school students have balanced diet. 12. Index of health practice of personal hygiene can be summarized as follows. A. A case of middle school students. 1) The percentage of health practice index in male and female was 49.6% and 48.1% respectively. Index of female students was higher than that of male students. 2) As for parent's occupation, public servants and company emplyee was upper level. Farming was low level. 3) As for income level, middle, level with 56.5% was highest in high income level and low level with 27.4% was highest in low income level. B. A case of high school students. 1) Middle level of health practice index was 46.0% of male students, upper and low level was 32.4% and 28.0% of female students respectively. 2) Middle level of health practice index was high in farming and company employee and upper level was high in commerce and service, low level with 60.0% was high in unemployed. 3) Upper practice index 35.7% appears in the rich and low practice index 38.3% appears in the poor. 13. Average points of Health practice about personal hygiene were as follows. (Full marks at 4). A. A case of middle school. Female (1.87 point) was higher than male (1.26 point). Night time (2.03 point) was higher than day time (1.66 point) and middle or small cities (2.17 point) are high than any other places. As for parent's occupation, students whose parents are company clerk get high marks (2.32) and ten students whose parent's job are service get next high marks (2.20). B. A case of high school. Female (1.53 point) was higher than male (1.22 point), as parents educational level were higher the point were higher, and as income level was higher, the points of health practice (1.78) were higher, and as for parents occupation, service get highest point (1.93) and commerce get next high point (1.86) public servant get low point (1.66). 14. The percentage of experience in smoking was 11.9% of middle school students and 60.9% of high school students. 15. The percentage of experience in inhalation of bond and administrating LSD was 4.3% of male middle school students, 8.4% of female middle school students, 6.9% of male high school students and 4.2% of female high school students. The knowledge level of communicable disease infection are very low in middle and high school students and practice level of personal hygiene are also very low. As a whole we can evaluate that middle and high school students are low level of health knowledge and practice. In conclusion, we must consider preparation for school health education program through establishing of health subjects in the carriculum, and securing of health education teachers and using materials and media program of health education. It is very important to establish macroscopic policy and strategy for public health education and to get people have right knowledge and practice for health.

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