• Title/Summary/Keyword: environmental changes

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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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A Study on Perception and Attitudes of Health Workers Towards the Organization and Activities of Urban Health Centers (도시보건소 직원의 보건소 업무에 대한 인식 및 견해)

  • Lee, Jae-Mu;Kang, Pock-Soo;Lee, Kyeong-Soo;Kim, Cheon-Tae
    • Journal of Yeungnam Medical Science
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    • v.12 no.2
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    • pp.347-365
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    • 1995
  • A survey was conducted to study perception and attitudes of health workers towards health center's activities and organization of health services, from August 15 to September 30, 1994. The study population was 310 health workers engaged in seven urban health centers in Taegu City area. A questionnaire method was used to collect data and response rate was 81.3 percent or 252 respondents. The following are summaries of findings: Profiles of study population: Health workers were predominantly female(62.3%); had college education(60.3%); and held medical and nursing positions(39.6%), technicians(30.6%) and public health/administrative positions(29.8%). Perceptions on health center's resources: Slightly more than a half(51.1%) of respondents expressed that physical facilities of the centers are inadequate; equipments needed are short(39.0%); human resource is inadequate(44.8%); and health budget allocated is insufficient(38.5%) to support the performance of health center's activities. Decentralization and health services: The majority revealed that the decentralization of government system would affect the future activities of health centers(51.9%) which may have to change. However, only one quarter of respondents(25.4%) seemed to view the decentralization positively as they expect that it would help perform health activities more effectively. The majority of the respondents(78.6%) insisted that the function and organization of the urban health centers should be changed. Target workload and job satisfaction: A large proportion (43.3%) of respondents felt that present target setting systems for various health activities are unrealistic in terms of community needs and health center's situation while only 11.1 percent responded it positively; the majority(57.5%) revealed that they need further training in professional fields to perform their job more effectively; more than one third(35.7%) expressed that they enjoy their professional autonomy in their job performance; and a considerable proportion (39.3%) said they are satisfied with their present work. Regarding the personnel management, more worker(47.3%) perceived it negatively than positive(11.5%) as most of workers seemed to think the personnel management practiced at the health centers is not fair or justly done. Health services rendered: Among health services rendered, health workers perceived the following services are most successfully delivered; they are, in order of importance, Tb control, curative services, and maternal and child health care. Such areas as health education, oral health, environmental sanitation, and integrated health services are needed to be strengthening. Regarding the community attitudes towards health workers, 41.3 percent of respondents think they are trusted by the community they serve. New areas of concern identified which must be included in future activities of health centers are, in order of priority, health care of elderly population, home health care, rehabilitation services, and such chronic diseases control programs as diabetes, hypertension, school health and mental health care. In conclusion, the study revealed that health workers seemed to have more negative perceptions and attitudes than positive ones towards organization and management of health services and activities performed by the urban health centers where they are engaged. More specifically, the majority of health workers studied revealed to have the following areas of health center's organization and management inadequate or insufficient to support effective performance of their health activities: Namely, physical facilities and equipments required are inadequate; human and financial resources are insufficient; personnel management is unsatisfactory; setting of service target system is unrealistic in terms of the community needs. However, respondents displayed a number of positive perceptions, particularly to those areas as further training needs and implementation of decentralization of government system which will bring more autonomy of local government as they perceived these change would bring the necessary changes to future activities of the health center. They also displayed positive perceptions in their job autonomy and have job satisfactions.

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Studies on Grain Filling and Quality Changes of Hard and Soft Wheat Grown under the Different Environmental Conditions (환경 변동에 따른 경ㆍ연질 소맥의 등숙 및 품질의 변화에 관한 연구)

  • Young-Soo Han
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.17
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    • pp.1-44
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    • 1974
  • These studies were made at Suwon in 1972 and at Suwon, Iri, and Kwangju in 1973 to investigate grain filling process and variation of grain quality of NB 68513 and Caprock as hard red winter wheat, Suke #169 as soft red winter wheat variety and Yungkwang as semi-hard winter variety, grown under-three different fertilizer levels and seeding dates. Other experiments were conducted to find the effects of temperature, humidity and light intensity on the grain filling process and grain quality of Yungkwang and NB 68513 wheat varieties. These, experiments were conducted at Suwon in 1973 and 1974. 1. Grain filling process of wheat cultivars: 1) The frequency distribution of a grain weight shows that wider distribution of grain weight was associated with large grain groups rather than small grain group. In the large grain groups, the frequency was mostly concentrated near mean value, while the frequency was dispersed over the values in the small grain group. 2) The grain weight was more affected by the grain thickness and width than by grain length. 3) The grain weight during the ripening period was rapidly increased from 14 days after flowering to 35 days in Yungkwang and from 14 days after flowering to 28 days in NB 68513. The large grain group, Yungkwang was rather slowly increased and took a longer period in increase of endosperm ratio of grain than the small grain group, NB 68513. 4) In general, the 1, 000 grain weight was reduced under high temperature, low humidity, while it was increased under low temperature and high humidity condition, and under high temperature and humidity condition. The effect of shading on grain weight was greater in high temperature than in low temperature condition and no definite tendency was found in high humidity condition. 5) The effects of temperature, humidity and shading on 1, 000 grain weight were greater in large-grain group, Yungkwang than in small grain group, NB 68513. Highly significant positive correlation was found between 1, 000 grain weight and days to ripening. 6) The 1, 000 grain weight and test weight were increased more or less as the fertilizer levels applied were increased. However, the rate of increasing 1, 000 grain weight was low when fertilizer levels were increased from standard to double. The 1, 000 grain weight was high when planted early. Such tendency was greater in Suwon than in Kwangju or Iri area. 2. Milling quality: 7) The milling rate in a same group of varieties was higher under the condition of low temperature, high humidity and early maturing culture which were responsible for increasing 1, 000 grain weight. No definite relations were found along with locations. 8) In the varieties tested, the higher milling rate was found in large grain variety, Yungkwang, and the lowest milling rate was obtained from Suke # 169, the small grain variety. But the small grained hard wheat variety such as Caprock and NB 68513 showed higher milling rate compared with the soft wheat variety, Suke # 169. 9) There were no great differences of ash content due to location, fertilizer level and seeding date while remarkable differences due to variety were found. The ash content was high in the hard wheat varieties such as NB 68513, Caprock and low in soft wheat varieties such as Yungkwang and Suke # 169. 3. Protein content: 10) The protein content was increased under the condition of high temperature, low humidity and shading, which were responsible for reduction of 1, 000 grain weight. The varietal differences of protein content due to high temperature, low humidity and shading conditions were greater in Yungkwang than in NB 68513. 11) The high content of protein in grain within one to two weeks after flowering might be due to the high ratio of pericarp and embryo to endosperm. As grains ripen, the effects of embryo and pericarp on protein content were decreased, reducing protein content. However, the protein content was getting increased from three or four weeks after flowering, and maximized at seven weeks after flowering. The protein content of grain at three to four weeks after flowering increased as the increase of 1, 000 grain weight. But the protein content of matured grain appeared to be affected by daily temperature on calender rather than by duration of ripening period. 12) Highly significant positive correlation value was found between the grain protein content and flour protein content. 13) The protein content was increased under the high level of fertilizers and late seeding. The local differences of protein content were greater in Suwon than in Kwangju and Iri. 14) Protein content in the varieties tested were high in Yungkwang, NB 68513 and Caprock, and low in Suke # 169. However, variation in protein content due to the cultural methods was low in Suke # 169. 15) Protein yield per unit area was increased in accordance with increase of fertilizer levels and early maturing culture. However, nitrogen fertilizer was utilized rather effectively in early maturing culture and Yungkwang was the highest in protein yield per unit area. 4. Physio-chemical properties of wheat flour: 16) Sedimentation value was higher under the conditions of high temperature, low humidity and high levels of fertilizers than under the conditions of low temperature, high moisture and low levels of fertilizers. Such differences of sedimentation values were more apparent in NB 68513 and Caprock than Yungkwang and Suke # 169. The local difference of sedimentation value was greater in Suwon than in Kwangju and Iri. Even though the sedimentation value was highly correlated with protein content of grain, the high humidity was considered one of the factors affecting sedimentation value. 17) Changes of Pelshenke values due to the differences of cultural practices and locations were generally coincident with sedimentation values. 18) The mixing time required for mixogram was four to six minutes in NB 68513, five to seven minutes in Cap rock. The great variation of mixing time for Yungkwang and Suke # 169 due to location and planting conditions was found. The mixing height and area were high in hard wheat than in soft wheat. Variation of protein content due to cultural methods were inconsistent. However, the pattern of mixogram were very much same regardless the treatments applied. With this regard, it could be concluded that the mixogram is a kind of method expressing the specific character of the variety. 19) Even though the milling property of NB 68513 and Caprock was deteriorated under either high temperature and low humidity of high fertilizer levels and late seeding conditions, baking quality was better due to improved physio-chemical properties of flour. In contrast, early maturing culture deteriorated physio-chemical properties, milling property of grain and grain protein yield per unit area was increased. However, it might be concluded that the hard wheat production of NB 68513 and Caprock for baking purpose could be done better in Suwon than in Iri or Kwangju area. 5. Interrelationships between the physio-chemical characters of wheat flour: 20) Physio-chemical properties of flour didn't have direct relationship with milling rate and ash content. Low grain weight produced high protein content and better physio-chemical flour properties. 21) In hard wheat varieties like NB 68513 and Caprock, protein content was significantly correlated with sedimentation value, Pelshenke value and mixing height. However, gluten strength and baking quality were improved by the increased protein content. In Yungkwang and Suk # 169, protein content was correlated with sedimentation value, but no correlations were found with Pelshenke value and mixing height. Consequently, increase of protein content didn't improve the gluten strength in soft wheat. 22) The highly significant relationships between protein content and gluten strength and sedimentation . value, and between Pelshenke value, mixogram and gluten strength indicated that the determination of mixogram and Pelshenke value are useful for de terming soft and hard type of varieties. Determination of sedimentation value is considered useful method for quality evaluation of wheat grain under different cultural practices.

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The Location and Landscape Composition of Yowol-pavilion Garden Interpreted from Tablet & Poetry (편액과 시문으로 본 요월정원림(邀月亭園林)의 입지 및 조영 해석)

  • Lee, Hyun-Woo;Kim, Sang-Wook;Ren, Qin-Hong
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.32 no.3
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    • pp.32-45
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    • 2014
  • The study attempts to interpret original location and landscape composition of Yowol-pavilion Garden under the premise that tablet and poetry are important criteria for inference of unique location and landscape composition of a pavilion garden. The study raises the meaning, status, and value of Yowol Pavilion Garden as a cultural asset. The results of the study are as follows. First, Yowol-pavilion Garden was a place where famous Confucius scholars in Joseon Dynasty in 16th Century, including Kim, Kyung-Woo, the owner of the garden, used to share the taste for the arts and poetries with their colleagues. Along with a main characteristic of Yowol Pavilion Garden as a hideout for the Confucius scholars who stayed away from a political turmoil, the new place characteristic of the garden, a bridgehead for the formation of regional identity, was discovered in the record of "Joseon-Hwanyeo-Seungram Honam-Eupji JangSeong-Eupji", As described in "The first creative poetry of Yowol-pavilion", the intention for the creation of Yowol-pavilion Garden and the motive for its landscape composition is interpreted as a space of rivalry where the world, reality and ideals are mixed up. Second, related to outstanding scenic factors and natural phenomena when taking a view from the pavilion, the name of the house 'Yowol', which means 'Greeting the moon rising on the Mt. Wolbong' is the provision of nature and taste for the arts, and is directly connected to the image of leaving the worldly. In other words, the name was identified to be the one that reflected the intention for landscape composition to follow the provision of nature separating from joy and sorrow of the mundane world. Third, as for the location, it was confirmed through "YeongGwang-Soksu-Yeoji-Seungram" that Yowol-pavilion Garden was a place where the person who made the pavilion prepared for relaxation after stepping down from a government post, and literature and various poetry show that it was also a place of outstanding scenic where Yellow-dragon River meandered facing Mt. Wolbong. Especially, according to an interview with a keeper, the visual perception frequency of the nightscape of Yowol-pavilion Garden is the highest when viewing by considering the east, the direction of Yellow-dragon River, as Suksigak[normal angle's view], towards Yowel-pavilion from the keeper's house. In addition, he said that the most beautiful landscape with high perception strength is when the moon came up from the left side of Yowol-pavilion, cuts across the Lagerstroemia india heal in front of Yowol-pavilion, and crosses the meridian between Mt. Wolbong peaks facing Yowol-pavilion. Currently, the exposure of Yowol-pavilion Garden is $SE\;141.2^{\circ}$, which is almost facing southeast. It is assumed that the exposure of Yowol-pavilion Garden was determined considering the optimized direction for appreciating the trace of the moon and the intention of securing the visibility as well as topographic conditions. Furthermore, it is presumed that the exposure of Yowol-pavilion Garden was determined so that the moon is reflected on the water of Yellow-dragon River and the moon and its reflection form a symmetry. Fourth, currently, Yowol-pavilion Garden is divided into 'inner garden sphere' composed of Yowol-pavilion, meeting place of the clan and administration building, and 'outer garden sphere' which is inclusive of entrance space, Crape Myrtle Community Garden and Pine Tree Forest in the back. Further, Yowol-pavilion Garden has been deteriorated as the edge was expanded to 'Small lake[Yong-so] and Gardens of aquatic plants sphere' and recently-created 'Yellow-dragon Pavilion and park sphere'. Fifth, at the time it was first made, Yowol-pavilion Garden was borderless gardens consisting of mountains and water taking a method of occupying a specific space of nearby nature centering around pavilion by embracing landscape viewed from the pavilion, but interpreted current complex landscapes are identified to be entirely different from landscapes of the original due to 'Different Changes', 'Fragmentation' and 'Apart piece' in many parts. Lastly, considering that Yowol-pavilion Garden belongs to the Cultural Properties Protection Zone, though not the restoration to the landscapes of the original described in tablet and literature record, at least taking a measure from the aspect of land use for minimizing adverse effect on landscape and visual damage is required.