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바이러스 벡터를 이용한 당뇨백서 간에서의 인슐린 유전자 발현
No full textDept. of Medical Science/박사[한글]
당뇨병을 치료하기 위하여 여러 가지 방법들이 시도되어 오고있으나 만족스럽지 못한 실정이다. 현재로서는 식후의 혈당을 조절하기 위한 방법과 기저치의 인슐린을 공급하는 방법을 적절하게 조합하여 시행되고 있다. 이 연구는 기저치의 인슐린을 제공하기 위하여 실행하고 있는 하루 두세 번 정도의 인슐린 주사법을 대신하여 유전자 요법으로 일정한 양의 인슐린을 공급하는 것을 목적으로 하였다. 구조적인 프로모터하에 퓨린에 의해 잘려지는 휴먼 프로인슐린을 아데노 바이러스와 아데노 수반 바이러스 벡터를 이용하여 스트렙토조토신에 의해 유도된 당뇨백서의 간문맥에 주입하고 혈당변화와 인슐린의 발현을 측정하였다. 아데노 바이러스를 이용한 유전자요법으로 혈액내의 인슐린농도를 증가시켜 당뇨백서에서 공복시의 고혈당을 개선하였고 아데노 수반 바이러스를 이용한 유전자 요법에 의해 3 개월이상 고혈당 상태에 있던 당뇨백서의 공복혈당이 정상으로 변화되었으며 당 분해능력과 당화 헤모글로빈 농도가 현저히 개선되었다. 유전자 요법이 실행된 백서에서 공복시의 혈중 인슐린의 농도 또한 일정하게 증가되어 기록되었다.
치료된 백서의 간에서 추출한 mRNA를 이용한 역전사 효소 중합효소 연쇄반응 (RT-PCR) 과 연쇄반응의 생산물의 뉴클레오타이드 분석을 통해 전달된 휴먼 프로인슐린 유전자의 발현을 확인하였다. 유전자 치료된 당뇨백서의 간조직과 췌장조직에서 면역조직염색법을 통하여 스트렙토조토신으로 췌도가 대부분 파괴되었음을 확인하였고 간에서 생성된 인슐린에 의하여 혈당강하의 효과가 나타났음을 보여주었다. 치료된 백서의 여러 가지 조직에서 추출한 RNA를 가지고 노던 블러팅을 실행한 결과 주입된 유전자의 발현이 간 조직에서 이루어졌음을 보여주었다. 이상의 결과로 구조적인 프로모터를 이용한 유전자 요법의 실행은 식후 혈당의 변화를 조절하기에는 제한적이었으나 일정한 양의 인슐린을 발현하고 공급함으로써 공복 상태의 고혈당을 개선하고 혈중 인슐린의 양을 증가시키는 것에 성공적인 결과를 보여주었다. 또한 아데노 수반 바이러스벡터를 이용한 유전자 치료법은 한번의 주입으로 오랜 기간동안 외부 인슐린 유전자를 발현함으로써 안정적이고 비교적 안전한 유전자 치료법임을 시사하였다.
[영문]A lot of therapeutic approaches to cure diabetes have been tried but still remains unsatisfied. The present treatments for diabetes involve frequent monitoring of blood glucose and life-long insulin injection or alternate combination with diabetic medicine. This research purposed to express furin-cleavage human proinsulin in liver of diabetic animal using adenovirus and adeno-associated virus (AAV) and to provide constant insulin level for basal requirement in diabetes. Adenovirus and adeno-associated virus expressing furin-cleavage human proinsulin (AAV-FhPI) under CMV promoter were administered into portal vein of streptozotocin (STZ)-induced diabetic rats. These resulted in the decrease of blood glucose levels within 24 hr after adenovirus injection and within 3 day after AAV injection respectively. In adenovirus-FhPI treated rats, serum insulin levels were increased and glucose tolerance was significantly improved although these effects lasted for less than 3 weeks. For a long-term insulin gene expression, AAV-FhPI was administered in STZ-induced diabetic rats and the treated rats exhibited the normalized blood glucose levels in fasting state over 3 months. The ability of glucose clearance approximated to non-diabetic control and the hemoglobin A1c concentrations were decreased in AAV-FhPI treated rats. Immunohistochemical studies of pancreas in adenovirus-FhPI treated rats and AAV-FhPI treated rats revealed the pancreatic β-cells were mostly destroyed while adenovirus-FhPI and AAV-FhPI transduced hepatocytes produced human insulin. In conclusion, viral vector-mediated furin-cleavage proinsulin gene expression using the constitutive promoter is feasible gene therapy to supply basal insulin, lessening the frequency of insulin injection. And portal vein injection is suitable to enhance the exogenous gene delivery into liver. Furthermore, the therapeutic potential of this approach would be realized through the study on regulating insulin secretion, for example, regulating promoter-based gene expression with glucose
insulin responsible elements.ope
Stress response assessment of coral under environmental changes using microarray
No full textThe Kuroshio is a warm current running from tropical Philippines through subtropical Taiwan to the temperate region of Japan and Korea and it warmed most rapidly in 1981–1998, when sea surface temperatures rose by 1.5°C (0.9°C/decade), almost 7 times the global rate. Coral of Taiwan and Japan were closely linked by the Kuroshio Current and inevitably, the coral population and its distribution in Korea could be affected by warming currents. In this study, we chose Eleutherobia rubra which is a azooxanthellate soft coral around Korean peninsula to investigate its gene expression change against the environmental stress and constructed microarray. The probes in microarray was based on the noble and known genes sequences from NGS technology. Approximately 75,000 genes from E. rubra were used and we carried out heat stress experiment using microarray. This study focused on the identification of functional genes in coral to assess the molecular level responses to environmental stress like as seawater temperature change. To assess the coral health condition as well as coral stress responses, we purposed to identify molecular level stress responses using gene expression change as well as functional genes responding to external stress like increased seawater temperature.2
Going Beyond Counting First Authors in Author Co-citation Analysis
Get PDFThe present study examines one of the fundamental aspects of author co-citation analysis (ACA) - the way co-citation
counts are defined. Co-citation counting provides the data on which all subsequent statistical analyses and mappings
are based, and we compare ACA results based on two different types of co-citation counting - the traditional type that
only counts the first one among a cited work's authors on the one hand and a non-traditional type that takes into
account the first 5 authors of a cited work on the other hand. Results indicate that the picture produced through this non-traditional author co-citation counting contains more coherent author groups and is therefore considerably clearer. However, this picture represents fewer specialties in the research field being studied than that produced through the traditional first-author co-citation counting when the same number of top-ranked authors is selected and analyzed. Reasons for these effects are discussed
Assessing Thermal Stress Responses of Alveopora Japonica Using Microarray
No full textThe reef building corals in the region have rapidly expanded their range northward in response to warming temperatures. Furthermore, coral reefs of Taiwan and Japan were closely linked by the Kuroshio Current and inevitably, the coral population and its distribution in Korea could be affected by warming currents, similar to the situation in Japan and Taiwan. In this study, we chose Alveopora japonicawhich is a hard coral rapidly expanding the population around Jeju Island to investigate its gene expression change against the environmental stress and constructed microarray. The probes in microarray was based on the noble and known genes sequences from NGS technology. Approximately 60,000 genes from Alveopora japonica were used and we carried out heat stress experiment using microarray. This study focused on the identification of functional genes in coral to assess the molecular level responses to environmental stress like as seawater temperature change. Climate change and rising seawater temperature are placing spatially divergent stresses on the world's coral ecosystem through increasing ocean surface temperatures and ocean acidification. To assess the coral health condition as well as coral stress responses, we purposed to identify molecular level stress responses using gene expression change as well as functional genes responding to external stress like increased seawater temperature. Approximately 200 genes including Kinesin-like protein, guanine-nucleotide releasing factor, caspase, apoptotic cysteine protease, etc changed their gene expressions after heat shock stress. These isolated genes were clustered into functional groups like information storage and processing, cellular processes and signaling and metabolism. Over 70 % of isolated genes were related to the signal transduction.1
Bacterial Community Composition change in Temperate Octocoral, Scleronephthya gracillimum Responding to Season and Heat Stress
No full textCoral diversity is affected by climate change. Environmental impacts can induce the change of relationship between coral and its symbiotic microbial community. Furthermore the microbial community change could lead the coral diseases considered one of the reasons of coral death and the opportunistic infections in corals exposed to the increased temperature. In this study we collected a temperate octocoral, Scleronephthya gracillimum in Jeju,Korea and exposed to various seawater temperature (26, 28 and 30ºC ) to compare the composition of bacterial community using the Next Generation Sequencing technique. The results showed total 88 species of bacteria were found in S.gracillimum in wild condition and they were classified to 72 genus, 41 family, 32 order, 16 class and 13 phylum. Soft coral, S.gracillimum were enriched in OTUs from the families Hahellaceae, Mycoplasmataceae, Alteromonadaceae, Anaplasmataceae, and Rhodobacteraceae. The number of bacteria species belong to following 8 families, Flavobacteriaceae, Bacillaceae, Comamonadaceae, Alteromonadaceae, Pseudoalteromonadaceae, Hahellaceae, Pseudomonadaceae and Vibrionaceae robustly increased in responses to the heat stress in 26, 28 and 30ºC groups and the species Mesoflavibacter sabulilitoris, Vibrio tubiashii, Pseudomonas azotoformans, Oceanospirillum beijerinckii, Neptuniibacter Caesariensis, and Amphritea spongicola showed proportionally increase by temperature. The number of Endozoicomonas elysicola showed an increase up to 26ºC then decrease in 28 and 30ºC groups.1
Variations on the Author
Get PDF“Variations on the Author” discusses two of Eduardo Coutinho’s recent films (Um Dia na Vida, from 2010, and Últimas Conversas, posthumously released in 2015) and their contribution to the general question of documentary authorship. The director’s filmography is characterized by a consistent yet self-effacing form of authorial self-inscription: Coutinho often features as an interviewer that rather than express opinions propels discourses; an interviewer that is good at listening. This mode of self-inscription characterizes him as an author who is not expressive but who is nonetheless markedly present on the screen. In Um Dia na Vida, however, Coutinho is completely absent form the image, while Últimas Conversas, on the contrary, includes a confessional prologue that moves the director from the margins to the center of his films. This article examines the ways in which these works stand out in the filmography of a director who offers new insights into the notion of cinematic authorship
Seasonal gene expression changes and transcriptional responses of azooxanthellate octocoral, Eleutherobia rubra to heat shock
No full textAzooxanthellate octocoral, Eleutherobia rubra were collected in South Sea, Republic of Korea where was designated as a marine protected area. The Seas around Korean peninsula are at the edge of the Northwest Pacific Ocean and affected by the Kuroshio Warm Current (1). This current was highest in 1981-1998, when the seawater surface temperature rose 1.5 °C and over 10 years it rose 0.9 °C, 7 times higher than the Earth's surface temperature rise (2). The corals were exposed to the higher temperature than its habitual temperature and RNAs were extracted to investigate the differentially expressed genes and the transcriptional changes using microarray. Additionally we investigated the seasonal and regional gene expressions of E. rubra using RNA de novo sequencing. The investigation of regulatory mechanisms governing stress responses by monitoring the changes of transcriptional profiles can be early warning molecular markers and provide an important source of information on acclimatization or adaptation to external stresses. This study purposedto investigate the gene expressions changes of the temperate azooxanthellate octocoral responding to thermal shock and transcriptional regulation behind physiological resilience and acclimatization capabilities of E. rubra in seasonal seawater temperature changes.1
Appropriate Similarity Measures for Author Cocitation Analysis
Get PDFWe provide a number of new insights into the methodological discussion about author cocitation analysis. We first argue that the use of the Pearson correlation for measuring the similarity between authors’ cocitation profiles is not very satisfactory. We then discuss what kind of similarity measures may be used as an alternative to the Pearson correlation. We consider three similarity measures in particular. One is the well-known cosine. The other two similarity measures have not been used before in the bibliometric literature. Finally, we show by means of an example that our findings have a high practical relevance.information science;Pearson correlation;cosine;similarity measure;author cocitation analysis
Dispelling the Myths Behind First-author Citation Counts
Get PDFWe conducted a full-scale evaluative citation analysis study of scholars in the XML research field to explore just how different from each other author rankings resulting from different citation counting methods actually are, and to demonstrate the capability of emerging data and tools on the Web in supporting more realistic citation counting methods. Our results contest some common arguments for the continued
use of first-author citation counts in the evaluation of scholars, such as high correlations between author rankings by first-author citation counts and other citation
counting methods, and high costs of using more realistic citation counting methods that are not well-supported by the ISI databases. It is argued that increasingly available digital full text research papers make it possible for citation analysis studies to go beyond what the ISI databases have directly supported and to employ more
sophisticated methods
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