In contrast to the human lutropin receptor (hLHR) and rat LHR (rLHR), very few naturally occurring mutants in other mammalian species have been identified. The present study aimed to delineate the mechanism of signal transduction by three constitutively activating mutants (designated M410T, L469R, and D590Y) and two inactivating mutants (D383N and Y546F) of the eel LHR, known to be naturally occurring in human LHR transmembrane domains. The mutants were constructed and measured cyclic adenosine monophosphate (cAMP) accumulation via homogeneous time-resolved fluorescence assays in Chinese hamster ovary (CHO)-K1 cells. The activating mutant cells expressing eel LHR-M410T, L469R, and D590Y exhibited a 4.0-, 19.1-, and 7.8-fold increase in basal cAMP response without agonist treatment, respectively. However, inactivating mutant cells expressing D417N and Y558F did not completely impaired signal transduction. Specifically, signal transduction in the cells expressing activating mutant L469R was not occurred with a further ligand stimulation, showing that the maximal response exhibited approximately 53% of those of wild type receptor. Our results suggested that the constitutively activating mutants of the eel LHR consistently occurred without agonist treatment. These results provide important information of LHR function in fish and regulation with regard to mutations of highly conserved amino acids in glycoprotein hormone receptors.
Kim, B. K.;H. J. Chung;Park, J. H.;J. H. Woo;Park, M. Y.;H. H. Seong;W. K. Chang
Proceedings of the KSAR Conference
/
2003.06a
/
pp.69-69
/
2003
Although effect of TGF$\beta$$_1$ on preimplantation embryo development was reported at mice, little information relevant to this subject is known in bovine. The objectives of this study were to investigate TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors type I and II expression, known as important factors in the embryo development, at unfertilized oocytes and fertilized embryos that will be used as basic data to be compared to NT embryos. We postulated that TGF$\beta$$_1$ may have a beneficial effect on the preimplantation embryo and show different expression patterns as embryo stages change. We have used immunocytochemistry to investigate the presence in unfertilized oocytes and preimplantation embryos of TGF$\beta$$_1$ and the essential components of the TGF$\beta$$_1$ signalling pathway, TGF$\beta$$_1$ receptors type I and II. We found that both receptors, as well as TGF$\beta$$_1$, were present in the unfertilized oocytes. This indicates that TGF$\beta$$_1$, is a maternally expressed protein. At the morulae and blastocyst stages the TGF$\beta$$_1$ receptor type II was not present, but the TGF$\beta$$_1$ receptor type I was present at both stages and we can confirm the TGF$\beta$$_1$ expression of high level at 8-cell stage. These findings support our hypothesis that the TGF$\beta$$_1$, and TGF$\beta$$_1$ receptors may interact with the oocyte and preimplantation embryo, and that TGF$\beta$$_1$ signalling may be important for the development of the oocyte and the preimplahtation embryo.
Epigenetic writers including DNA and histone lysine methyltransferases (DNMT and HKMT, respectively) play an initiative role in the differentiation and development of eukaryotic organisms through the spatiotemporal regulation of functional gene expressions. However, the epigenetic mechanisms have long been suspected in helminth parasites lacking the major DNA methyltransferases DNMT1 and DNMT3a/3b. Very little information on the evolutionary status of the epigenetic tools and their role in regulating chromosomal genes is currently available in the parasitic trematodes. We previously suggested the probable role of a DNMT2-like protein (CsDNMT2) as a genuine epigenetic writer in a trematode parasite Clonorchis sinensis. Here, we analyzed the phylogeny of HKMT subfamily members in the liver fluke and other platyhelminth species. The platyhelminth genomes examined conserved genes for the most of SET domain-containing HKMT and Disruptor of Telomeric Silencing 1 subfamilies, while some genes were expanded specifically in certain platyhelminth genomes. Related to the high gene dosages for HKMT activities covering differential but somewhat overlapping substrate specificities, variously methylated histones were recognized throughout the tissues/organs of C. sinensis adults. The temporal expressions of genes involved in eggshell formation were gradually decreased to their lowest levels proportionally to aging, whereas those of some epigenetic tool genes were re-boosted in the later adult stages of the parasite. Furthermore, these expression levels were significantly affected by treatment with DNMT and HKMT inhibitors. Our data strongly suggest that methylated histones are potent epigenetic markers that modulate the spatiotemporal expressions of C. sinensis genes, especially those involved in sexual reproduction.
Glycoprotein hormones have a common $\alpha$-subunit that is involved in the signaling pathway together with G protein, adenylcyclase and cAMP induction; however, it is an unclear how this common structure is related to hormonal action. To determine the biological functions of the COOH-terminal amino acids in the $\alpha$-subunit of these glycoprotein hormones, a tethered-molecule was constructed by fusing the $NH_2$-terminus of the $\alpha$-subunit to the COOH-terminus of the $\beta$-subunit of equine chorionic gonadotropin (eCG). The following deletion mutants were created by PCR; Ile was inserted at position 96 to form ${\Delta}96$, Lys was substituted at position 95 to form ${\Delta}95$, His was inserted at position 93 to form ${\Delta}93$ and Tyr was substituted at position 87 to form ${\Delta}87$. Each mutant was transfected into CHO-K1 cells. Tethered-wt eCG, and ${\Delta}96$, ${\Delta}95$, and ${\Delta}93$ mutants were efficiently secreted into the medium but the ${\Delta}87$ mutant was not secreted. Interestingly, the RT-PCR, real-time PCR, and northern blot analyses confirmed that the RNA was transcribed in the ${\Delta}87$ mutant. However, the ${\Delta}87$ mutant protein was not detected in the medium or the intracellular fraction of the cell lysates. The LH- and FSH-like activities of the recombinant proteins were assayed in terms of cAMP production using rat LH/CG and rat FSH receptors. The metabolic clearance rate (MCR) was determined by injecting rec-eCG (2 IU) into the tail vein. The ${\Delta}95$ and ${\Delta}93$ mutants were completely inactive in both the LH- and FSH-like activity assays. The ${\Delta}96$ mutant showed slight activity in the LH-like activity assay. In comparison to the wild type, the activity of the ${\Delta}96$ mutant in the FSH-like activity assay was the highest among all the mutants. The MCR assay in which rec-eCG was injected showed a peak at 10 min in all the treatment groups, which disappeared 4 h after injection. These results imply a direct interaction between the receptor and the COOH-terminal region of the a-subunit. The data also reveal a significant difference in the mechanism by which the eCG hormone interacts with the rLH and rFSH receptors. The COOH-terminal region of the $\alpha$-subunit is very important for the secretion and functioning of this hormone.
Kim, Min-Jee;Lee, Hyoung-Song;Kang, Inn-Soo;Jeong, Seon-Yong;Kim, Hyon-J.
Journal of Genetic Medicine
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v.7
no.2
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pp.125-132
/
2010
Purpose: Preimplantation genetic diagnosis (PGD), also known as embryo screening, is a pre-pregnancy technique used to identify genetic defects in embryos created through in vitro fertilization. PGD is considered a means of prenatal diagnosis of genetic abnormalities. PGD is used when one or both genetic parents has a known genetic abnormality; testing is performed on an embryo to determine if it also carries the genetic abnormality. The main advantage of PGD is the avoidance of selective pregnancy termination as it imparts a high likelihood that the baby will be free of the disease under consideration. The application of PGD to genetic practices, reproductive medicine, and genetic counseling is becoming the key component of fertility practice because of the need to develop a custom PGD design for each couple. Materials and Methods: In this study, a survey on the contents of genetic counseling in PGD was carried out via direct contact or e-mail with the patients and specialists who had experienced PGD during the three months from February to April 2010. Results: A total of 91 persons including 60 patients, 49 of whom had a chromosomal disorder and 11 of whom had a single gene disorder, and 31 PGD specialists responded to the survey. Analysis of the survey results revealed that all respondents were well aware of the importance of genetic counseling in all steps of PGD including planning, operation, and follow-up. The patient group responded that the possibility of unexpected results (51.7%), genetic risk assessment and recurrence risk (46.7%), the reproduction options (46.7%), the procedure and limitation of PGD (43.3%) and the information of PGD technology (35.0%) should be included as a genetic counseling information. In detail, 51.7% of patients wanted to be counseled for the possibility of unexpected results and the recurrence risk, while 46.7% wanted to know their reproduction options (46.7%). Approximately 96.7% of specialists replied that a non-M.D. genetic counselor is necessary for effective and systematic genetic counseling in PGD because it is difficult for physicians to offer satisfying information to patients due to lack of counseling time and specific knowledge of the disorders. Conclusions: The information from the survey provides important insight into the overall present situation of genetic counseling for PGD in Korea. The survey results demonstrated that there is a general awareness that genetic counseling is essential for PGD, suggesting that appropriate genetic counseling may play a important role in the success of PGD. The establishment of genetic counseling guidelines for PGD may contribute to better planning and management strategies for PGD.
Korean Journal of Agricultural and Forest Meteorology
/
v.16
no.4
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pp.368-383
/
2014
Climate change is expected to affect population density, phenology, distribution, morphological traits, reproduction and genetics of insects, and even in the extinction of insects. To develop novel research subjects for predicting climate change effect, basic information about biological and ecological data on insect species should be compiled and reviewed. For this reason, this study was conducted to collect the biological information on insect pests that are essential for predicting potential damage caused by insect pests in future environment. In addition, we compared domestic and foreign research trends regarding climate change effect and suggested future research subjects. Domestic researchers were rather narrow in the subject, and were mostly conducted based on short-term monitoring data to determine relationship between insects and environmental variables. On the other hand, foreign researches studied on various subjects to analyze the effect of climate change, such as changes in distribution of insect using long-term monitoring data or their prediction using population parameters and models, and monitoring of the change of the insect community structure. To determine change of the phenology, distribution, overwintering characteristics, and genetic structures of insects under climate change through development of monitoring technique, in conclusion, further researches are needed. Also, development of population models for major or potential pests is important for prediction of climate change effects.
Tscherskia triton is widely distributed in Northern China, Korea, and the adjacent areas of Russia. Except its distribution, reproduction, and growth development related to life history, reproductive cycle and reproductive organs of T. triton are rarely studied in Korea. The purpose of this study was characterized the estrous cycle of T. triton captured in Jeju Island in order to provide information to a better information of captive breeding of the species when long-day (16L : 8D) and short-day (8L : 16D) photoperiod. Then, histological study of the ovaries and uterus with five females in each photoperiod was performed. The duration of the estrus cycle was 4~5 days and it showed regular cycle pattern. Results of the vaginal cytology examination showed four characteristic phase of the estrous cycle in long-day photoperiod (16L : 8D): proestrus, estrus, metestrus and diestrus. However, in short-day photoperiod, the diestrus phage of the estrus cycle was maintained from the $6^{th}$ to $12^{th}$ day. In the long-day photoperiod, females had many Graafian follicles and corpus luteums in large ovaries, and developed uterine glands in the thick endometrium. But they had some primary, secondary and tertiary follicles, and undeveloped uterine glands in the thin endometrium during short-day photoperiod. These results were identified difference of the estrus cycle and histological characteristics of reproductive tracts according to the photoperiod. These results are very important clues to the reproductive biology of T. triton, and it will be widely used as date for maintaining biodiversity.
This paper addresses questions on the marginalized position of youths of 'Gangbuk' and elucidates how they construct their own identities in the individual trajectories of everyday life. Three years of research, including participatory observation and in-depth interviews, was conducted on nine students from Northeastern district of Seoul. The research also adopted reflexive photography interview method in order to encourage the informants to actively participate in the research. The result illustrates the diversity of the everyday life experiences. More 'marginalized' youths from middle to lower class background had to endure the burdens of their daily lives without programs. Still, they were elaborating their own cultural taste and positive self-narratives at the periphery of the mainstream culture, by practicing music, online community activities and bodily performances. They had to negotiate the crucial turn of life after their graduation, when they entered into the harsh social competition with limited resources. We observed how they gradually assimilate the identity of the 'working youth', some of them developing a positive valorization of their experiences labor. Findings underline the active role of the cultural practices in the making of class identity of the youth and the necessity of researches situating the making of class identity and the reproduction of the class for the youth in the larger geography of class culture in the contemporary Korean society. Lastly, it is argued that these youths should not be considered as determined subjects, who reproduce already established class identities, but as active agents of their lives who deserve more respects and attentions from the society.
Throughout developing digital technology, reproduction of image is growing better day by day. And at the same time, diverse image editing softwares are developed to manage images easily. In the process of editing images, those programs could delete or modify EXIF files which have the original image information; therefore images without the origin source are widely spread on the web site after editing. This matter could affect analysis of images due to the distortion of originality. Especially in the court of law, the source of evidence should be expressed clearly; therefore digital image EXIF file without deletion or distortion could not be the objective evidence. In this research, we try to trace the identification of a digital camera in order to solve digital images originality, and also we focus on lens distortion correction algorism which is used in digital image processing. Lens distortion correction uses mapping algorism, and at this moment it also uses interpolation algorism to prevent aliasing artifact and reconstruction artifact. At this point interpolation shows the similar mapping pattern; therefore we want to find out the interpolation evidence. We propose a minimum filter algorism in order to detect interpolation pattern and adjust the same minimum filter coefficient in two areas; one has interpolation and the second has no interpolation. Throughout DFT, we confirm frequency character between each area. Based on this result, we make the final detection map by using differences between two areas. In other words, thereby the area which has the interpolation caused by mapping is adjusted using minimum filter for detection algorism; the second area which has no interpolation tends to different frequency character.
Today many countries consider the culture and technology industry as a high value-added industry, and make intensive investments into cultural content for its growth. In the culture and technology industry, the OSMU (One Source Multi Use) business structure, which creates content using diverse media based on one original work and thus generates a high added value, has received much attention. Recently Korea has also witnessed an increase in the cases in which the cartoon medium is converted into movies or drama. Cartoons are now put to the most use as an original source, in which the process of active secondary reproduction and recreation are taking place. In the process of recreating original works, media characteristics function as an important variable. Stories are newly reproduced by the detailed information (form) characteristics of each medium. Despite those developments, however, there are not as many active comparative studies on media-converted works as there are actual productions of them, which calls for a need to more fully activate specific comparisons of cases. By comparing and investigating original works and secondarily recreated ones through media conversion, the study made comparisons between the cartoon and movie medium and described the common and different features of the two media in terms of detailed information characteristics. In order to provide a specific case, it also analyzed the cartoon Tajja and movie Tajja through media conversion, focusing on 'conversion of narrative structure.'
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