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An Experimental and Numerical Study on the Survivability of a Long Pipe-Type Buoy Structure in Waves (긴 파이프로 이뤄진 세장형 부이 구조물의 파랑 중 생존성에 관한 모형시험 및 수치해석 연구)

  • Kwon, Yong-Ju;Nam, Bo-Woo;Kim, Nam-Woo;Park, In-Bo;Kim, Sea-Moon
    • Journal of Navigation and Port Research
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    • v.42 no.6
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    • pp.427-436
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    • 2018
  • In this study, experimental and numerical analysis were performed on the survivability of a long pipe-type buoy structure in waves. The buoy structure is an articulated tower consisting of an upper structure, buoyancy module, and gravity anchor with long pipes forming the base frame. A series of experiment were performed in the ocean engineering basin of KRISO with the scaled model of 1/ 22 to evaluate the survivability of the buoy structure at West Sea in South Korea. Survival condition was considered as the wave of 50 year return period. Additional experiments were performed to investigate the effects of current and wave period. The factors considered for the evaluation of the buoy's survival were the pitch angle of the structure, anchor reaction force, and the number of submergence of the upper structure. Numerical simulations were carried out with the OrcaFlex, the commercial program for the mooring analysis, with the aim of performing mutual validation with the experimental results. Based on the evaluation, the behavior characteristics of the buoy structure were first examined according to the tidal conditions. The changes were investigated for the pitch angle and anchor reaction force at HAT and LAT conditions, and the results directly compared with those obtained from numerical simulation. Secondly, the response characteristics of the buoy structure were studied depending on the wave period and the presence of current velocity. Third, the number of submergence through video analysis was compared with the simulation results in relation to the submergence of the upper structure. Finally, the simulation results for structural responses which were not directly measured in the experiment were presented, and the structural safety discussed in the survival waves. Through a series of survivability evaluation studies, the behavior characteristics of the buoy structure were examined in survival waves. The vulnerability and utility of the buoy structure were investigated through the sensitivity studies of waves, current, and tides.

On the Origin and Development of Iconography of the Twelve Zodiac Signs of Royal Tombs of Joseon Dynasty (조선왕릉 십이지신상(十二支神像)의 도상(圖像) 원류와 전개 과정)

  • Kim, Ji Yeon
    • Korean Journal of Heritage: History & Science
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    • v.42 no.4
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    • pp.198-221
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    • 2009
  • Royal tombs of Joseon dynasty are the crystallization of history, ideology, culture, art, architecture, and ritual ceremony of Joseon dynasty, all mingled in one. So, they are very significant symbols showing 500 years of dynastic history as a whole. Among various factors comprising a royal tomb, stone figures surrounding grave mound are special factors as a symbol protecting it. Further among them, twelve zodiac images arrayed nearest to the grave mound represent the core of the function. Images of twelve zodiac signs originated from the tombs of the Unified Silla Kingdom are certain to hold important role and position in the construction of royal tombs, judging from huge scale and excellent sculptural art of them. However, both their scale and form had been gradually simplified in Goryeo and Joseon dynasty, thus the importance of them has been underestimated compared to other stone figures Images of twelve zodiac signs were very important factors which decorated royal tombs both as a protective role and as a concept of direction. Their historicity and symbolism cannot be neglected in that they had been transmitted to the royal tombs of Joseon dynasty. In this paper, images of twelve zodiac signs expressed in the royal tombs of Joseon dynasty are classified into 3 forms, and reviewed the origin and development of them for each period. They could be classified into 3 forms ; civil vassals with human body and head, civil vassals with human body and head wearing hat decorated with zodiac animals, Chinese characters of either zodiac signs or either a combination of 10 calendar signs and 8 trigrams. The above 3 forms originated from China and became a favorite motif to decorate the royal tombs from early Joseon period until late Joseon by replacing each other and thus changing along the course of the dynasty. In the meantime, we can see a unique character in the images of twelve zodiac signs of royal tombs of Joseon dynasty. In some cases, 24 directions are expressed in which 10 calender signs and 8 trigrams are composed altogether. Images of twelve zodiac signs in the royal bombs of Joseon dynasty are very significant as evidences by which we can confirm uniqueness and tradition of Korean tomb system transmitted from Unified Silla period.

Numerical Analysis of Pile Foundation Considering the Thawing and Freezing Effects (융해-동결작용을 고려한 말뚝 기초에 관한 수치해석 연구)

  • Park, Woo-Jin ;Park, Dong-Su;Shin, Mun-Beom;Seo, Young-Kyo
    • Journal of the Korean Geotechnical Society
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    • v.39 no.5
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    • pp.51-63
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    • 2023
  • Numerical analysis was conducted to determine the effect of soil behavior by thawing and freezing of seasonal frozen soil on pile foundations. The analysis was performed using the finite element method (FEM) to simulate soil-pile interaction based on the atmosphere temperature change. Thermomechanical coupled modeling using FEM was applied with the temperature-dependent nonlinear properties of the frozen soil. The analysis model cases were applied to the MCR and HDP models to simulate the elastoplastic behavior of soil. The numerical analysis results were analyzed and compared with various conditions having different length and width sizes of the pile. The results of the numerical analysis showed t hat t he HDP model was relat ively passive, and t he aspect and magnit ude of t he bearing capacit y and displacement of the pile head were similar depending on the length and width of the pile conditions. The vertical displacement of the pile head by thawing and freezing of the ground showed a large variation in displacement for shorter length conditions. In the MCR model, the vertical displacement appeared in the maximum thaw settlement and frost heaving of 0.0387 and 0.0277 m, respectively. In the HDP model, the vertical displacement appeared in the maximum thaw settlement and frost heaving of 0.0367 and 0.0264 m, respectively. The results of the pile bearing capacity for the two elastoplastic models showed a larger difference in the width condition than the length condition of the pile, with a maximum of about 14.7% for the width L condition, a maximum of about 5.4% for M condition, and a maximum of about 5.3% for S condition. The significance of the effect on the displacement of the pile head and the bearing capacity depended on the pile-soil contact area, and the difference depended on the presence or absence of an active layer in the soil and its thickness.

Analysis of Clothing in a Painting Album of a 60th Wedding Anniversary Feast in the Collection of the National Museum of Korea (국립중앙박물관 소장 《회혼례도첩》 속 등장인물의 복식 고찰)

  • LEE Eunjoo
    • Korean Journal of Heritage: History & Science
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    • v.56 no.3
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    • pp.76-98
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    • 2023
  • The results of analyzing the outfits of male and female characters depicted in the "Hoehonryedocheop" (回婚禮圖帖, Deoksu 6375) held by the National Museum of Korea and estimating its production date of the "Hoehonryedocheop"are as follows. Firstly, an elderly groom is depicted wearing a patterned heukdanryeong (黑團領) with rank badges, a garment commonly donned by government officials in works such as "Jeonanryedo (奠鴈禮圖)" and "Gyobaeryedo (交拜禮圖)". And the old groom in "Heonsuryedo (獻壽禮圖)" "Jeobbindo (接賓圖)" and "Jungroeyeondo (重牢宴圖)" is shown wearing a jarip (purple hat) without a paeyoung (ornamental jewelry strap), accompanied by jade colored robe with a red strap belt. Gireokabeom (雁夫) is observed wearing a jarip (紫笠) adorned with a paeyoung (貝纓) and a patternless heukdanryeong with rank badges. Adult male descendants are depicted wearing dopo (道袍), while guests wear dopo, cheolrik (帖裏), and jikryeong (直領), accompanied by red and blue straps denoting their social status. Jingssi (徵氏), sidong (侍童), and young grandchildren are observed wearing jungchimak(中赤莫). The young servants are wearing jungchimak, and the boys carrying the food have braided their hair and worn sochangu (小氅衣), while adults servants wore jeonrip (氈笠) and sochangui. Performers are seen clad in a sochangui, jeonbok, and a blue sash around their waists. Secondly, the elderly bride is portrayed wearing a keunmeori (ceremonial headdress) and a green wonsam (圓衫) in "Gyobaeryedo," while in "Heonsuyeondo," she is depicted in a blue skirt and a jade colored jeogori (赤古里). Women descendants are shown adorning headdress decorations, such as binyeo(簪), banja(斑子) and pearl daenggi (眞珠唐紒) on their eoyeomeori (於于味, ceremonial headdress). They are further dressed in skirts of navy, red, and jade hues, paired with various-colored jeogori. Additionally, a woman wearing a navy skirt and a green jangot (長衣) is also depicted. The dongnyeo (童女, unmarried women) wear beolsaengmeri (娘子雙髻), headdress) with long binyeo and long dodaik-daenggi (都多益唐只). They wear chilbo-jokduri (七寶簇頭里) and a red skirt with a green hoejang-jeogori (回裝赤古里). Bija (婢子) wears garima (加里亇) on her eoyeomeori and is seen dressed in skirts and jeogori resembling those worn by noble women, albeit with lighter colors, shorter skirt length, and a subdued volume. Ginyeo's attire bears similarities to that of noble women, although with a dress with less vibrant tones and devoid of decorations on the eoyeomeori. Thirdly, based on the main character's jarip, along with the cheolrik and jikryeong worn by the guests, as well as the performances by musicians of the military camp, it is suggested that the main character of the 60th wedding anniversary is connected to the Ministry of Military Affairs or the military camp. Judging by the military band's short-sleeved vest, the silhouettes of the women dress, and the headdresses, it is likely that the "Hoehonryedocheop" was produced between the 1760s and 1780s.

Attitude Confidence and User Resistance for Purchasing Wearable Devices on Virtual Reality: Based on Virtual Reality Headgears (가상현실 웨어러블 기기의 구매 촉진을 위한 태도 자신감과 사용자 저항 태도: 가상현실 헤드기어를 중심으로)

  • Sohn, Bong-Jin;Park, Da-Sul;Choi, Jaewon
    • Journal of Intelligence and Information Systems
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    • v.22 no.3
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    • pp.165-183
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    • 2016
  • Over the past decade, there has been a rapid diffusion of technological devices and a rising number of various devices, resulting in an escalation of virtual reality technology. Technological market has rapidly been changed from smartphone to wearable devices based on virtual reality. Virtual reality can make users feel real situation through sensing interaction, voice, motion capture and so on. Facebook.com, Google, Samsung, LG, Sony and so on have investigated developing platform of virtual reality. the pricing of virtual reality devices also had decreased into 30% from their launched period. Thus market infrastructure in virtual reality have rapidly been developed to crease marketplace. However, most consumers recognize that virtual reality is not ease to purchase or use. That could not lead consumers to positive attitude for devices and purchase the related devices in the early market. Through previous studies related to virtual reality, there are few studies focusing on why the devices for virtual reality stayed in early stage in adoption & diffusion context in the market. Almost previous studies considered the reasons of hard adoption for innovative products in the viewpoints of Typology of Innovation Resistance, MIR(Management of Innovation Resistant), UTAUT & UTAUT2. However, product-based antecedents also important to increase user intention to purchase and use products in the technological market. In this study, we focus on user acceptance and resistance for increasing purchase and usage promotions of wearable devices related to virtual reality based on headgear products like Galaxy Gear. Especially, we added a variables like attitude confidence as a dimension for user resistance. The research questions of this study are follows. First, how attitude confidence and innovativeness resistance affect user intention to use? Second, What factors related to content and brand contexts can affect user intention to use? This research collected data from the participants who have experiences using virtual rality headgears aged between 20s to 50s located in South Korea. In order to collect data, this study used a pilot test and through making face-to-face interviews on three specialists, face validity and content validity were evaluated for the questionnaire validity. Cleansing the data, we dropped some outliers and data of irrelevant papers. Totally, 156 responses were used for testing the suggested hypotheses. Through collecting data, demographics and the relationships among variables were analyzed through conducting structural equation modeling by PLS. The data showed that the sex of respondents who have experience using social commerce sites (male=86(55.1%), female=70(44.9%). The ages of respondents are mostly from 20s (74.4%) to 30s (16.7%). 126 respondents (80.8%) have used virtual reality devices. The results of our model estimation are as follows. With the exception of Hypothesis 1 and 7, which deals with the two relationships between brand awareness to attitude confidence, and quality of content to perceived enjoyment, all of our hypotheses were supported. In compliance with our hypotheses, perceived ease of use (H2) and use innovativeness (H3) were supported with its positively influence for the attitude confidence. This finding indicates that the more ease of use and innovativeness for devices increased, the more users' attitude confidence increased. Perceived price (H4), enjoyment (H5), Quantity of contents (H6) significantly increase user resistance. However, perceived price positively affect user innovativeness resistance meanwhile perceived enjoyment and quantity of contents negatively affect user innovativeness resistance. In addition, aesthetic exterior (H6) was also positively associated with perceived price (p<0.01). Also projection quality (H8) can increase perceived enjoyment (p<0.05). Finally, attitude confidence (H10) increased user intention to use virtual reality devices. however user resistance (H11) negatively affect user intention to use virtual reality devices. The findings of this study show that attitude confidence and user innovativeness resistance differently influence customer intention for using virtual reality devices. There are two distinct characteristic of attitude confidence: perceived ease of use and user innovativeness. This study identified the antecedents of different roles of perceived price (aesthetic exterior) and perceived enjoyment (quality of contents & projection quality). The findings indicated that brand awareness and quality of contents for virtual reality is not formed within virtual reality market yet. Therefore, firms should developed brand awareness for their product in the virtual market to increase market share.

Studies on Dry Matter Yields , Chemical Composition and Net Energy Accumulation in Three Leading Temperate Grass Species I. Influence of meteorolgical factors on the dry matter productivity and net energy value under different cutting management (주요 북방형목초의 건물수량 , 화학성분 및 Net Energy 축적에 관한 연구 I. 기상환경 및 예취관리에 따른 건물 및 에너지 생산성 변화)

  • F. Muhlschlegel;G. Voigtlander
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.6 no.2
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    • pp.103-110
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    • 1986
  • The experiments were carried out to study the influence of meteorological factors and cutting management on dry matter accumulation and net energy value in orchardgrass (Dactlylis glomerata L.) cv. Potomac and Baraula, perennial ryegrass (Lolium perenne L.) cv. Reveille and Semperweide and meadow fescue (Festuca pratensis Huds.) cv. Cosmos 11 and N.F.G.. The field trials were designed as a split plot design with three cutting regimes of 6-7 cuts at grazing stage, 4-5 cuts at silage stage and 3 cuts at hat stage in Korea and West Germany from 1975 to 1979. The results obtained are summarized as follows: 1. Productivity of orchardgrass, perennial ryegrass and meadow fescue were mainly affected by cutting systems and meteorological factors, especially air temperature, rainfalls, solar radiation and their interactions. In West Germany, cutting frequency was to be found asan most important factor influenced to dry matter yield and net energy value. 2. Orchardgrass, taken as average of all experimental sites in Korea, produced high yield of 875 kg/10 a in dry matter, which was as much as 32% and 27% higher than those of perennial ryegrass and meadow fescue, respectively. The annual dry matter yields of orchardgrass from 1976 to 1977 were shown a little variation. Dry matter yields in Freising and Braunschweig in West Germany were increased in all grass species continuously. 3. Orchardgrass, perennial ryegrass and meadow fescue showed different response to cutting frequency. The highest dry matter yields were found under 3 cuts at hay stage for orchardgrass and 4-5 cuts at silage stage for perennial ryegrass and meadow fescue. In West Germany, dry matter yields, as average of all grass species under different cutting systems, were 1326 kg, 1175 kg and 1098 kg/10a for 3 cuts, 4-5 cuts and 6-7 cuts, respectively. 4. Chemical composition and net energy concentration of temperate grasses were influenced by cutting managements. The highest yields of digestible crude protein were obtained under 6-7 cuts at grazing stage both in Korea and West Germany. In net energy yields, 3 cutting system produced the highest yield with 694 (orchardgrass), 665 (perennial ryegrass) an 623 kStE/10 a (meadow fescue). However, frequent cutting at grazing and silage stage produced higher yields than 3 cuts at hay stage in Cheju, Suweon and Taekwalyong.

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Dynamic Limit and Predatory Pricing Under Uncertainty (불확실성하(不確實性下)의 동태적(動態的) 진입제한(進入制限) 및 약탈가격(掠奪價格) 책정(策定))

  • Yoo, Yoon-ha
    • KDI Journal of Economic Policy
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    • v.13 no.1
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    • pp.151-166
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    • 1991
  • In this paper, a simple game-theoretic entry deterrence model is developed that integrates both limit pricing and predatory pricing. While there have been extensive studies which have dealt with predation and limit pricing separately, no study so far has analyzed these closely related practices in a unified framework. Treating each practice as if it were an independent phenomenon is, of course, an analytical necessity to abstract from complex realities. However, welfare analysis based on such a model may give misleading policy implications. By analyzing limit and predatory pricing within a single framework, this paper attempts to shed some light on the effects of interactions between these two frequently cited tactics of entry deterrence. Another distinctive feature of the paper is that limit and predatory pricing emerge, in equilibrium, as rational, profit maximizing strategies in the model. Until recently, the only conclusion from formal analyses of predatory pricing was that predation is unlikely to take place if every economic agent is assumed to be rational. This conclusion rests upon the argument that predation is costly; that is, it inflicts more losses upon the predator than upon the rival producer, and, therefore, is unlikely to succeed in driving out the rival, who understands that the price cutting, if it ever takes place, must be temporary. Recently several attempts have been made to overcome this modelling difficulty by Kreps and Wilson, Milgram and Roberts, Benoit, Fudenberg and Tirole, and Roberts. With the exception of Roberts, however, these studies, though successful in preserving the rationality of players, still share one serious weakness in that they resort to ad hoc, external constraints in order to generate profit maximizing predation. The present paper uses a highly stylized model of Cournot duopoly and derives the equilibrium predatory strategy without invoking external constraints except the assumption of asymmetrically distributed information. The underlying intuition behind the model can be summarized as follows. Imagine a firm that is considering entry into a monopolist's market but is uncertain about the incumbent firm's cost structure. If the monopolist has low cost, the rival would rather not enter because it would be difficult to compete with an efficient, low-cost firm. If the monopolist has high costs, however, the rival will definitely enter the market because it can make positive profits. In this situation, if the incumbent firm unwittingly produces its monopoly output, the entrant can infer the nature of the monopolist's cost by observing the monopolist's price. Knowing this, the high cost monopolist increases its output level up to what would have been produced by a low cost firm in an effort to conceal its cost condition. This constitutes limit pricing. The same logic applies when there is a rival competitor in the market. Producing a high cost duopoly output is self-revealing and thus to be avoided. Therefore, the firm chooses to produce the low cost duopoly output, consequently inflicting losses to the entrant or rival producer, thus acting in a predatory manner. The policy implications of the analysis are rather mixed. Contrary to the widely accepted hypothesis that predation is, at best, a negative sum game, and thus, a strategy that is unlikely to be played from the outset, this paper concludes that predation can be real occurence by showing that it can arise as an effective profit maximizing strategy. This conclusion alone may imply that the government can play a role in increasing the consumer welfare, say, by banning predation or limit pricing. However, the problem is that it is rather difficult to ascribe any welfare losses to these kinds of entry deterring practices. This difficulty arises from the fact that if the same practices have been adopted by a low cost firm, they could not be called entry-deterring. Moreover, the high cost incumbent in the model is doing exactly what the low cost firm would have done to keep the market to itself. All in all, this paper suggests that a government injunction of limit and predatory pricing should be applied with great care, evaluating each case on its own basis. Hasty generalization may work to the detriment, rather than the enhancement of consumer welfare.

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A Report on the Shigella Cultures Isolated in Korea(1971) (1971년(年) 한국(韓國)에서 분리(分離)한 이질균(痢疾菌)에 관(關)한 보고(報告))

  • Ryu, Young-Hat;Kim, Soon-Hee
    • The Journal of the Korean Society for Microbiology
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    • v.7 no.1
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    • pp.9-15
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    • 1972
  • The authors identified 45 Shigella cultures among 63 suspectable cultures obtained from the cases showing dysentery or the like symptoms in various parts of the country during the period from February to September, 1971. Of 45 cultures, 36 cultures belonged to Subgroup B, 7 cultures to Subgroup C and 2 cultures to Subgroup D. There was none of cultures belonging to Subgroup A in 1971, although the authors detected one culture of that among 41 identified Shigella in 1967. Of 36 cultures belonging to Subgroup B, 27 cultures were $B_{2a}$ and 3 cultures of each $B_{3c},\;B_{4a}$ and $B_y$ were serotyped, respectively. Of 7 cultures beloning to Subgroup C, one was $C_{11}$ and other six cultures were $C_{15}$ and all S. boydii were isolated in Seoul area. It might be possible to suggest that there was a tendency of decreasing prevalence of S. dysenteriae and of increasing prevalence of S. boydii in Korea, although the number of cultures. tested were very much limited to conclude, if it was compared with the results obtained by the authors in 1967 that the fourty-one identified Shigella were composed of one culture of S. dysenteriae, 36 cultures of S. flexneri, 4 cultures of S. sonnei and none of S. boydii. According to the results obtained from the biochemical tests, the positive ratio of Indol tests in Subgroup B was 31/36, which could be higher than 9/36 observed in 1961, but which was nearly correlated with the result published by Ewing and his colleagues in U.S.A. The positive rates of both sorbitol and raffinose were lower than that observed by Ewing and his colleagues in S. flexneri. Regarding with the sensitivity of Shigella cultures to the antibiotics being widely used in the country, 46.2% were sensitive to chloramphenicol, which was very much higher than that observed by Park, and 85.8% were sensitive to ampicillin, which would be the drug of choice according to the results from the In Vitro tests.

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APPROXIMATE ESTIMATION OF RECRUITMENT IN FISH POPULATION UTILIZING STOCK DENSITY AND CATCH (밀도지수와 어획량으로서 수산자원의 가입량을 근사적으로 추정하는 방법)

  • KIM Kee Ju
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.8 no.2
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    • pp.47-60
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    • 1975
  • For the calculation of population parameter and estimation of recruitment of a fish population, an application of multiple regression method was used with some statistical inferences. Then, the differences between the calculated values and the true parameters were discussed. In addition, this method criticized by applying it to the statistical data of a population of bigeye tuna, Thunnus obesus of the Indian Ocean. The method was also applied to the available data of a population of Pacific saury, Cololabis saira, to estimate its recuitments. A stock at t year and t+1 year is, $N_{0,\;t+1}=N_{0,\;t}(1-m_t)-C_t+R_{t+1}$ where $N_0$ is the initial number of fish in a given year; C, number o: fish caught; R, number of recruitment; and M, rate of natural mortality. The foregoing equation is $$\phi_{t+1}=\frac{(1-\varrho^{-z}{t+1})Z_t}{(1-\varrho^{-z}t)Z_{t+1}}-\frac{1-\varrho^{-z}t+1}{Z_{t+1}}\phi_t-a'\frac{1-\varrho^{-z}t+1}{Z_{t+1}}C_t+a'\frac{1-\varrho^{-z}t+1}{Z_{t+1}}R_{t+1}......(1)$$ where $\phi$ is CPUE; a', CPUE $(\phi)$ to average stock $(\bar{N})$ in number; Z, total mortality coefficient; and M, natural mortality coefficient. In the equation (1) , the term $(1-\varrho^{-z}t+1)/Z_{t+1}$s almost constant to the variation of effort (X) there fore coefficients $\phi$ and $C_t$, can be calculated, when R is a constant, by applying the method of multiple regression, where $\phi_{t+1}$ is a dependent variable; $\phi_t$ and $C_t$ are independent variables. The values of Mand a' are calculated from the coefficients of $\phi_t$ and $C_t$; and total mortality coefficient (Z), where Z is a'X+M. By substituting M, a', $Z_t$, and $Z_{t+1}$ to the equation (1) recruitment $(R_{t+1})$ can be calculated. In this precess $\phi$ can be substituted by index of stock in number (N'). This operational procedures of the method of multiple regression can be applicable to the data which satisfy the above assumptions, even though the data were collected from any chosen year with similar recruitments, though it were not collected from the consecutive years. Under the condition of varying effort the data with such variation can be treated effectively by this method. The calculated values of M and a' include some deviation from the population parameters. Therefore, the estimated recruitment (R) is a relative value instead of all absolute one. This method of multiple regression is also applicable to the stock density and yield in weight instead of in number. For the data of the bigeye tuna of the Indian Ocean, the values of estimated recruitment (R) calculated from the parameter which is obtained by the present multiple regression method is proportional with an identical fluctuation pattern to the values of those derived from the parameters M and a', which were calculated by Suda (1970) for the same data. Estimated recruitments of Pacific saury of the eastern coast of Korea were calculated by the present multiple regression method. Not only spring recruitment $(1965\~1974)$ but also fall recruitment $(1964\~1973)$ was found to fluctuate in accordance with the fluctuations of stock densities (CPUE) of the same spring and fall, respectively.

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Sesquiterpenoids Bioconversion Analysis by Wood Rot Fungi

  • Lee, Su-Yeon;Ryu, Sun-Hwa;Choi, In-Gyu;Kim, Myungkil
    • 한국균학회소식:학술대회논문집
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    • 2016.05a
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    • pp.19-20
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    • 2016
  • Sesquiterpenoids are defined as $C_{15}$ compounds derived from farnesyl pyrophosphate (FPP), and their complex structures are found in the tissue of many diverse plants (Degenhardt et al. 2009). FPP's long chain length and additional double bond enables its conversion to a huge range of mono-, di-, and tri-cyclic structures. A number of cyclic sesquiterpenes with alcohol, aldehyde, and ketone derivatives have key biological and medicinal properties (Fraga 1999). Fungi, such as the wood-rotting Polyporus brumalis, are excellent sources of pharmaceutically interesting natural products such as sesquiterpenoids. In this study, we investigated the biosynthesis of P. brumalis sesquiterpenoids on modified medium. Fungal suspensions of 11 white rot species were inoculated in modified medium containing $C_6H_{12}O_6$, $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ for 20 days. Cultivation was stopped by solvent extraction via separation of the mycelium. The metabolites were identified as follows: propionic acid (1), mevalonic acid lactone (2), ${\beta}$-eudesmane (3), and ${\beta}$-eudesmol (4), respectively (Figure 1). The main peaks of ${\beta}$-eudesmane and ${\beta}$-eudesmol, which were indicative of sesquiterpene structures, were consistently detected for 5, 7, 12, and 15 days These results demonstrated the existence of terpene metabolism in the mycelium of P. brumalis. Polyporus spp. are known to generate flavor components such as methyl 2,4-dihydroxy-3,6-dimethyl benzoate; 2-hydroxy-4-methoxy-6-methyl benzoic acid; 3-hydroxy-5-methyl phenol; and 3-methoxy-2,5-dimethyl phenol in submerged cultures (Hoffmann and Esser 1978). Drimanes of sesquiterpenes were reported as metabolites from P. arcularius and shown to exhibit antimicrobial activity against Gram-positive bacteria such as Staphylococcus aureus (Fleck et al. 1996). The main metabolites of P. brumalis, ${\beta}$-Eudesmol and ${\beta}$-eudesmane, were categorized as eudesmane-type sesquiterpene structures. The eudesmane skeleton could be biosynthesized from FPP-derived IPP, and approximately 1,000 structures have been identified in plants as essential oils. The biosynthesis of eudesmol from P. brumalis may thus be an important tool for the production of useful natural compounds as presumed from its identified potent bioactivity in plants. Essential oils comprising eudesmane-type sesquiterpenoids have been previously and extensively researched (Wu et al. 2006). ${\beta}$-Eudesmol is a well-known and important eudesmane alcohol with an anticholinergic effect in the vascular endothelium (Tsuneki et al. 2005). Additionally, recent studies demonstrated that ${\beta}$-eudesmol acts as a channel blocker for nicotinic acetylcholine receptors at the neuromuscular junction, and it can inhibit angiogenesis in vitro and in vivo by blocking the mitogen-activated protein kinase (MAPK) signaling pathway (Seo et al. 2011). Variation of nutrients was conducted to determine an optimum condition for the biosynthesis of sesquiterpenes by P. brumalis. Genes encoding terpene synthases, which are crucial to the terpene synthesis pathway, generally respond to environmental factors such as pH, temperature, and available nutrients (Hoffmeister and Keller 2007, Yu and Keller 2005). Calvo et al. described the effect of major nutrients, carbon and nitrogen, on the synthesis of secondary metabolites (Calvo et al. 2002). P. brumalis did not prefer to synthesize sesquiterpenes under all growth conditions. Results of differences in metabolites observed in P. brumalis grown in PDB and modified medium highlighted the potential effect inorganic sources such as $C_4H_{12}N_2O_6$, $KH_2PO_4$, $MgSO_4$, and $CaCl_2$ on sesquiterpene synthesis. ${\beta}$-eudesmol was apparent during cultivation except for when P. brumalis was grown on $MgSO_4$-free medium. These results demonstrated that $MgSO_4$ can specifically control the biosynthesis of ${\beta}$-eudesmol. Magnesium has been reported as a cofactor that binds to sesquiterpene synthase (Agger et al. 2008). Specifically, the $Mg^{2+}$ ions bind to two conserved metal-binding motifs. These metal ions complex to the substrate pyrophosphate, thereby promoting the ionization of the leaving groups of FPP and resulting in the generation of a highly reactive allylic cation. Effect of magnesium source on the sesquiterpene biosynthesis was also identified via analysis of the concentration of total carbohydrates. Our current study offered further insight that fungal sesquiterpene biosynthesis can be controlled by nutrients. To profile the metabolites of P. brumalis, the cultures were extracted based on the growth curve. Despite metabolites produced during mycelia growth, there was difficulty in detecting significant changes in metabolite production, especially those at low concentrations. These compounds may be of interest in understanding their synthetic mechanisms in P. brumalis. The synthesis of terpene compounds began during the growth phase at day 9. Sesquiterpene synthesis occurred after growth was complete. At day 9, drimenol, farnesol, and mevalonic lactone (or mevalonic acid lactone) were identified. Mevalonic acid lactone is the precursor of the mevalonic pathway, and particularly, it is a precursor for a number of biologically important lipids, including cholesterol hormones (Buckley et al. 2002). Farnesol is the precursor of sesquiterpenoids. Drimenol compounds, bi-cyclic-sesquiterpene alcohols, can be synthesized from trans-trans farnesol via cyclization and rearrangement (Polovinka et al. 1994). They have also been identified in the basidiomycota Lentinus lepideus as secondary metabolites. After 12 days in the growth phase, ${\beta}$-elemene caryophyllene, ${\delta}$-cadiene, and eudesmane were detected with ${\beta}$-eudesmol. The data showed the synthesis of sesquiterpene hydrocarbons with bi-cyclic structures. These compounds can be synthesized from FPP by cyclization. Cyclic terpenoids are synthesized through the formation of a carbon skeleton from linear precursors by terpene cyclase, which is followed by chemical modification by oxidation, reduction, methylation, etc. Sesquiterpene cyclase is a key branch-point enzyme that catalyzes the complex intermolecular cyclization of the linear prenyl diphosphate into cyclic hydrocarbons (Toyomasu et al. 2007). After 20 days in stationary phase, the oxygenated structures eudesmol, elemol, and caryophyllene oxide were detected. Thus, after growth, sesquiterpenes were identified. Per these results, we showed that terpene metabolism in wood-rotting fungi occurs in the stationary phase. We also showed that such metabolism can be controlled by magnesium supplementation in the growth medium. In conclusion, we identified P. brumalis as a wood-rotting fungus that can produce sesquiterpenes. To mechanistically understand eudesmane-type sesquiterpene biosynthesis in P. brumalis, further research into the genes regulating the dynamics of such biosynthesis is warranted.

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