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Integrated Rotary Genetic Analysis Microsystem for Influenza A Virus Detection

  • Jung, Jae Hwan;Park, Byung Hyun;Choi, Seok Jin;Seo, Tae Seok
    • Proceedings of the Korean Vacuum Society Conference
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    • 2013.08a
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    • pp.88-89
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    • 2013
  • A variety of influenza A viruses from animal hosts are continuously prevalent throughout the world which cause human epidemics resulting millions of human infections and enormous industrial and economic damages. Thus, early diagnosis of such pathogen is of paramount importance for biomedical examination and public healthcare screening. To approach this issue, here we propose a fully integrated Rotary genetic analysis system, called Rotary Genetic Analyzer, for on-site detection of influenza A viruses with high speed. The Rotary Genetic Analyzer is made up of four parts including a disposable microchip, a servo motor for precise and high rate spinning of the chip, thermal blocks for temperature control, and a miniaturized optical fluorescence detector as shown Fig. 1. A thermal block made from duralumin is integrated with a film heater at the bottom and a resistance temperature detector (RTD) in the middle. For the efficient performance of RT-PCR, three thermal blocks are placed on the Rotary stage and the temperature of each block is corresponded to the thermal cycling, namely $95^{\circ}C$ (denature), $58^{\circ}C$ (annealing), and $72^{\circ}C$ (extension). Rotary RT-PCR was performed to amplify the target gene which was monitored by an optical fluorescent detector above the extension block. A disposable microdevice (10 cm diameter) consists of a solid-phase extraction based sample pretreatment unit, bead chamber, and 4 ${\mu}L$ of the PCR chamber as shown Fig. 2. The microchip is fabricated using a patterned polycarbonate (PC) sheet with 1 mm thickness and a PC film with 130 ${\mu}m$ thickness, which layers are thermally bonded at $138^{\circ}C$ using acetone vapour. Silicatreated microglass beads with 150~212 ${\mu}L$ diameter are introduced into the sample pretreatment chambers and held in place by weir structure for construction of solid-phase extraction system. Fig. 3 shows strobed images of sequential loading of three samples. Three samples were loaded into the reservoir simultaneously (Fig. 3A), then the influenza A H3N2 viral RNA sample was loaded at 5000 RPM for 10 sec (Fig. 3B). Washing buffer was followed at 5000 RPM for 5 min (Fig. 3C), and angular frequency was decreased to 100 RPM for siphon priming of PCR cocktail to the channel as shown in Figure 3D. Finally the PCR cocktail was loaded to the bead chamber at 2000 RPM for 10 sec, and then RPM was increased up to 5000 RPM for 1 min to obtain the as much as PCR cocktail containing the RNA template (Fig. 3E). In this system, the wastes from RNA samples and washing buffer were transported to the waste chamber, which is fully filled to the chamber with precise optimization. Then, the PCR cocktail was able to transport to the PCR chamber. Fig. 3F shows the final image of the sample pretreatment. PCR cocktail containing RNA template is successfully isolated from waste. To detect the influenza A H3N2 virus, the purified RNA with PCR cocktail in the PCR chamber was amplified by using performed the RNA capture on the proposed microdevice. The fluorescence images were described in Figure 4A at the 0, 40 cycles. The fluorescence signal (40 cycle) was drastically increased confirming the influenza A H3N2 virus. The real-time profiles were successfully obtained using the optical fluorescence detector as shown in Figure 4B. The Rotary PCR and off-chip PCR were compared with same amount of influenza A H3N2 virus. The Ct value of Rotary PCR was smaller than the off-chip PCR without contamination. The whole process of the sample pretreatment and RT-PCR could be accomplished in 30 min on the fully integrated Rotary Genetic Analyzer system. We have demonstrated a fully integrated and portable Rotary Genetic Analyzer for detection of the gene expression of influenza A virus, which has 'Sample-in-answer-out' capability including sample pretreatment, rotary amplification, and optical detection. Target gene amplification was real-time monitored using the integrated Rotary Genetic Analyzer system.

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Management Plan for Humanistic and Ecological Characteristics of Suweol Village Forest in Tongyoung (통영 수월숲의 인문학적 특성 및 생태적 특성을 고려한 관리방안)

  • Lim, Eui-Jea;Lee, Soo-Dong;Kim, Mi-Jeong
    • Korean Journal of Environment and Ecology
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    • v.27 no.1
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    • pp.85-98
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    • 2013
  • In order to propose effective conservation management plan, this study verified ecological characteristics, humanities and Social characteristics. The research site is private property which is owned belonging to the Kim's of Gimhae that have long history. The study site is more than a thousand years old and was created for protecting from typhoon. There held the religious ritual what is called Dongsinje until 1960s. There have been protected and managed by the villagers. As the results of analysis, the area of windbreak are $12,392.69m^2$. The windbreak is dominated old years deciduous broad-leaved tree such as Zelkova serrata, Celtis sinensis, Aphananthe aspera. Around there were farmlands(52.1%), urbanized area(26.3%), forest area(16.6%). The vegetation communities of windbreak were classified by considering the dominant species and current status of forest. The forest types are following as; A. aspera community(I) which is using less pressure, Platycarya strobilacea-Carpinus coreana-Z. serrata community(II). Z. serrata community(III) which is using high pressure, Z. serrata-A. aspera community(IV), Z. serrata community(V) which is damaged under canopy trees. The windbreak was in good condition whereas, there were concerns the some wrong status was being undermined such as the wrong forest restoration projects in the past, the trails that is penetrating inside the forest, building up education facilities. Therefore, in order to restore the value of windbreak what is so called Suwol forest, we should improve the problems of forest ecosystem such as wrong management, forest fragmentation by facilities and decline in forest by lack of growing the next generation trees. In addition, we should remove excessive resting facilities and lead to passive use of forest. to improve the way of wrong management, moreover, we should close off he trails that is penetrating inside the forest for improving fragmentation. We should restore vegetation restoration and fostering the next generation trees for forest ecosystem. In order to restore unique of histo-cultural and ecological forest landscape, the next generation trees should be grown up that is the dominant species in Suwol forest. Moreover, as a results of comparing the between good vegetation communities and damaged vegetation communities, it is necessary to complementary planting for demeged vegetation communities, therefore there needs to 10.8 under canopy trees, 79.7 shrubs.

A Comparative Study of Juvenile Black-faced Spoonbills Platalea Minor Home Range in Gujido and Chilsando Islets, South Korea (구지도, 칠산도 저어새 유조의 행동권 비교 연구)

  • Son, Seok-Jun;Kang, Jung-Hoon;Kwon, In-Ki;Kim, Dal-Ho;Lee, Ki-Sup;Yoo, Jeong-Chil
    • Korean Journal of Environment and Ecology
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    • v.34 no.2
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    • pp.99-105
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    • 2020
  • Migratory birds use a variety of breeding and wintering sites, and it is particularly important to understand more information on breeding and feeding sites for the conservation and management of endangered species. Black-faced spoonbills (Platalea minor) are an international endangered species distributed in East Asia. The majority of black-faced spoonbills breed on uninhabited islets off the west coast of the Korean Peninsula during the breeding season, and they are distributed in East Asia such as Taiwan, Hong Kong, southern China, Japan, and Jeju island during the winter season. In this study, we used a wild animal location tracking system to analyze and compare home ranges of three black-faced spoonbills spending the post-fledging stage in Gujido islet in Incheon and Chilsando islet in Yeonggwang each in 2015. The tree black-faced spoonbills in Guji islet showed a home range in coastal areas in Hwanghaenam-do and Gangneung-gun. The home range size (mean±SD) was estimated to be 425.49±116.95 ㎢ using 100% MCP, 43.61±18.51 ㎢ using KDE 95%, and 7.46±3.68 ㎢using KDE 50%. The tree black-faced spoonbills in Chilsando islet showed a home range in the Baeksu tidal flat and the Buan Saemangeum area with a size of 99.38±55.29 ㎢ using 100% MCP, 19.87±6.05 ㎢ using KDE 95%, and 1.16±0.53 ㎢ using KDE 50%. The figured indicated that the tree black-faced spoonbills breeding in Gujido islet had a wider home range than those breeding in Chilsando islet. During the post-fledging stage, the home ranges of black-faced spoonbills were mostly breeding in mudflats. Therefore, it is necessary to minimize human intervention, such as the construction of roads and structures and the human access, to protect the habitats during the period.

Showing Filial Piety: Ancestral Burial Ground on the Inwangsan Mountain at the National Museum of Korea (과시된 효심: 국립중앙박물관 소장 <인왕선영도(仁旺先塋圖)> 연구)

  • Lee, Jaeho
    • MISULJARYO - National Museum of Korea Art Journal
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    • v.96
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    • pp.123-154
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    • 2019
  • Ancestral Burial Ground on the Inwangsan Mountain is a ten-panel folding screen with images and postscripts. Commissioned by Bak Gyeong-bin (dates unknown), this screen was painted by Jo Jung-muk (1820-after 1894) in 1868. The postscripts were written by Hong Seon-ju (dates unknown). The National Museum of Korea restored this painting, which had been housed in the museum on separate sheets, to its original folding screen format. The museum also opened the screen to the public for the first time at the special exhibition Through the Eyes of Joseon Painters: Real Scenery Landscapes of Korea held from July 23 to September 22, 2019. Ancestral Burial Ground on the Inwangsan Mountain depicts real scenery on the western slopes of Inwangsan Mountain spanning present-day Hongje-dong and Hongeun-dong in Seodaemun-gu, Seoul. In the distance, the Bukhansan Mountain ridges are illustrated. The painting also bears place names, including Inwangsan Mountain, Chumohyeon Hill, Hongjewon Inn, Samgaksan Mountain, Daenammun Gate, and Mireukdang Hall. The names and depictions of these places show similarities to those found on late Joseon maps. Jo Jung-muk is thought to have studied the geographical information marked on maps so as to illustrate a broad landscape in this painting. Field trips to the real scenery depicted in the painting have revealed that Jo exaggerated or omitted natural features and blended and arranged them into a row for the purposes of the horizontal picture plane. Jo Jung-muk was a painter proficient at drawing conventional landscapes in the style of the Southern School of Chinese painting. Details in Ancestral Burial Ground on the Inwangsan Mountain reflect the painting style of the School of Four Wangs. Jo also applied a more decorative style to some areas. The nineteenth-century court painters of the Dohwaseo(Royal Bureau of Painting), including Jo, employed such decorative painting styles by drawing houses based on painting manuals, applying dots formed like sprinkled black pepper to depict mounds of earth and illustrating flowers by dotted thick pigment. Moreover, Ancestral Burial Ground on the Inwangsan Mountain shows the individualistic style of Jeong Seon(1676~1759) in the rocks drawn with sweeping brushstrokes in dark ink, the massiveness of the mountain terrain, and the pine trees simply depicted using horizontal brushstrokes. Jo Jung-muk is presumed to have borrowed the authority and styles of Jeong Seon, who was well-known for his real scenery landscapes of Inwangsan Mountain. Nonetheless, the painting lacks an spontaneous sense of space and fails in conveying an impression of actual sites. Additionally, the excessively grand screen does not allow Jo Jung-muk to fully express his own style. In Ancestral Burial Ground on the Inwangsan Mountain, the texts of the postscripts nicely correspond to the images depicted. Their contents can be divided into six parts: (1) the occupant of the tomb and the reason for its relocation; (2) the location and geomancy of the tomb; (3) memorial services held at the tomb and mysterious responses received during the memorial services; (4) cooperation among villagers to manage the tomb; (5) the filial piety of Bak Gyeong-bin, who commissioned the painting and guarded the tomb; and (6) significance of the postscripts. The second part in particular is faithfully depicted in the painting since it can easily be visualized. According to the fifth part revealing the motive for the production of the painting, the commissioner Bak Gyeongbin was satisfied with the painting, stating that "it appears impeccable and is just as if the tomb were newly built." The composition of the natural features in a row as if explaining each one lacks painterly beauty, but it does succeed in providing information on the geomantic topography of the gravesite. A fair number of the existing depictions of gravesites are woodblock prints of family gravesites produced after the eighteenth century. Most of these are included in genealogical records and anthologies. According to sixteenth- and seventeenth-century historical records, hanging scrolls of family gravesites served as objects of worship. Bowing in front of these paintings was considered a substitute ritual when descendants could not physically be present to maintain their parents' or other ancestors' tombs. Han Hyo-won (1468-1534) and Jo Sil-gul (1591-1658) commissioned the production of family burial ground paintings and asked distinguished figures of the time to write a preface for the paintings, thus showing off their filial piety. Such examples are considered precedents for Ancestral Burial Ground on the Inwangsan Mountain. Hermitage of the Recluse Seokjeong in a private collection and Old Villa in Hwagae County at the National Museum of Korea are not paintings of family gravesites. However, they serve as references for seventeenth-century paintings depicting family gravesites in that they are hanging scrolls in the style of the paintings of literary gatherings and they illustrate geomancy. As an object of worship, Ancestral Burial Ground on the Inwangsan Mountain recalls a portrait. As indicated in the postscripts, the painting made Bak Gyeong-bin "feel like hearing his father's cough and seeing his attitudes and behaviors with my eyes." The fable of Xu Xiaosu, who gazed at the portrait of his father day and night, is reflected in this gravesite painting evoking a deceased parent. It is still unclear why Bak Gyeong-bin commissioned Ancestral Burial Ground on the Inwangsan Mountain to be produced as a real scenery landscape in the folding screen format rather than a hanging scroll or woodblock print, the conventional formats for a family gravesite paintings. In the nineteenth century, commoners came to produce numerous folding screens for use during the four rites of coming of age, marriage, burial, and ancestral rituals. However, they did not always use the screens in accordance with the nature of these rites. In the Ancestral Burial Ground on the Inwangsan Mountain, the real scenery landscape appears to have been emphasized more than the image of the gravesite in order to allow the screen to be applied during different rituals or for use to decorate space. The burial mound, which should be the essence of Ancestral Burial Ground on the Inwangsan Mountain, might have been obscured in order to hide its violation of the prohibition on the construction of tombs on the four mountains around the capital. At the western foot of Inwangsan Mountain, which was illustrated in this painting, the construction of tombs was forbidden. In 1832, a tomb discovered illegally built on the forbidden area was immediately dug up and the related people were severely punished. This indicates that the prohibition was effective until the mid-nineteenth century. The postscripts on the Ancestral Burial Ground on the Inwangsan Mountain document in detail Bak Gyeong-bin's efforts to obtain the land as a burial site. The help and connivance of villagers were necessary to use the burial site, probably because constructing tombs within the prohibited area was a burden on the family and villagers. Seokpajeong Pavilion by Yi Han-cheol (1808~1880), currently housed at the Los Angeles County Museum of Art, is another real scenery landscape in the format of a folding screen that is contemporaneous and comparable with Ancestral Burial Ground on the Inwangsan Mountain. In 1861 when Seokpajeong Pavilion was created, both Yi Han-cheol and Jo Jung-muk participated in the production of a portrait of King Cheoljong. Thus, it is highly probable that Jo Jung-muk may have observed the painting process of Yi's Seokpajeong Pavilion. A few years later, when Jo Jungmuk was commissioned to produce Ancestral Burial Ground on the Inwangsan Mountain, his experience with the impressive real scenery landscape of the Seokpajeong Pavilion screen could have been reflected in his work. The difference in the painting style between these two paintings is presumed to be a result of the tastes and purposes of the commissioners. Since Ancestral Burial Ground on the Inwangsan Mountain contains the multilayered structure of a real scenery landscape and family gravesite, it seems to have been perceived in myriad different ways depending on the viewer's level of knowledge, closeness to the commissioner, or viewing time. In the postscripts to the painting, the name and nickname of the tomb occupant as well as the place of his surname are not recorded. He is simply referred to as "Mister Bak." Biographical information about the commissioner Bak Gyeong-bin is also unavailable. However, given that his family did not enter government service, he is thought to have been a person of low standing who could not become a member of the ruling elite despite financial wherewithal. Moreover, it is hard to perceive Hong Seon-ju, who wrote the postscripts, as a member of the nobility. He might have been a low-level administrative official who belonged to the Gyeongajeon, as documented in the Seungjeongwon ilgi (Daily Records of Royal Secretariat of the Joseon Dynasty). Bak Gyeong-bin is presumed to have moved the tomb of his father to a propitious site and commissioned Ancestral Burial Ground on the Inwangsan Mountain to stress his filial piety, a conservative value, out of his desire to enter the upper class. However, Ancestral Burial Ground on the Inwangsan Mountain failed to live up to its original purpose and ended up as a contradictory image due to its multiple applications and the concern over the exposure of the violation of the prohibition on the construction of tombs on the prohibited area. Forty-seven years after its production, this screen became a part of the collection at the Royal Yi Household Museum with each panel being separated. This suggests that Bak Gyeong-bin's dream of bringing fortune and raising his family's social status by selecting a propitious gravesite did not come true.