The purpose of this study is to evaluate an efficacy of in vitro differentiated human embryonic stem (hES, MB03) cells expressing Nurr1 in relief of symptomatic motor behavior of Parkinson's disease (PD) animal models MB03 was genetically modified to express Nurr1 protein and was induced to differentiate according to 2-/4+ protocol using retinoic acid and ascorbic acid. The differentiation-induced cells were selected for 10 to 20 days thereafter in N2 medium. Upon selection, cells expressing GFAP, TH, or NF200 were 38.8%, 11%, and 20.5%, respectively. in order to examine therapeutic effects of the differentiated cells in PD animal model, rats were unilaterally lesioned by administration of 6-kydroxydopamine HCI (6-OHDA) into medial forebrain region (MFB, AP -4.4 mm, ML 1.2 mm, DV 78 mm with incision bar set at -2.4 mm), as a reference to bregma and the surface of the skull. Confirmation of successful lesion by apomorphine-induced rotational behavior, differentiated cells were transplanted into the striatum (AP 1.0, ML 3.5, DV -5.0; AP 0.6, ML 2.5, DV -4.5). Improvements of asymmetric motor behavior by the transplantation were examined every two weeks after the surgery. In two weeks, numbers of rotation by the experimental rats were (P<0.05) of the number before transplantation, however, the ratio increased slightly to in six weeks. In contrast, the ratio of sham-grafted animals ranged from 112.3+8.5% to 139.2+28.9% during the examination. Immunohistochemical studies further confirmed the presence, survival, migration, and expression of TH of the transplanted human cells.
Embryonic stem (ES) cells proliferate extensively in the undifferentiated state and have the potential to differentiate into a variety of cell types in response to various environmental cues. The generation of functional dopaminergic neurons from ES cells is promising for cell replacement therapy to treat Parkinson's disease. We compared the in vitro differentiation potential of pluripotent human embryonic stem (hES, MB03) cells induced with basic fibroblast growth factor (bFGF) or retinoic acid (RA). Both types of treatment resulted in similar neural cell differentiation patterns at the terminal differentiation stage, specifically, 75% neurons and 11% glial cells. Additionally, treatment of hES cells with brain derived neurotrophic factor (BDNF) or transforming growth factor (TGF)- during the terminal differentiation stage led to significantly increased tyrosine hydroxylase (TH) expression, compared to control (P<0.05). In contrast, no effect was observed on the rate of mature or glutamic acid decarboxylase-positive neurons. Immunostaining and HPLC analyses revealed the higher levels of TH (20.3%) and dopamine in bFGF and TGF- treated hES cells than in RA or BDNF treated hES cells. The results indicate that TGF- may be successfully used in the bFGF induction protocol to yield higher numbers of functional dopaminergic neurons from hES cells.
Embryonic stem (ES) cells, derived from preimplantation embryos, are able to differentiate into various types of cells consisting the whole body, or pluripotency. In addition to the plasticity, ES cells are expected to be different from terminally differentiated cells in very many ways, such as patterns of gene expressions, ability and response of the cells in confronting environmental stimulations, metabolism, and growth rate. As a model system to differentiate these two types of cells, human ES (hES, MB03) cells and terminally differentiated cells (HeLa), we examined the ability of these two types of cells in confronting a severe oxidative insult, that is . Ratio of dying cells as determined by the relative amount of dye neutral red entrapped within the cells after the exposures. Cell death rates were not significantly different when either MB03 or HeLa were exposed up to 0.4 mM . However, relative amount of dye entrapped within the cells sharply decreased down to 0.12% in HeLa cells when the cells were exposed to 0.8 mM , while it was approximately 54% in MB03. Pretreatment of cells with BSO (GSH chelator) and measurement of GSH content results suggest that cellular GSH is the major defensive mechanism of hES cells. Induction of apoptosis in hES cell was confirmed by DNA laddering, induction of Bax, and chromatin condensation. In summary, hES cells 1) are extremely resistant to oxidative stress, 2) utilize GSH as a major defensive mechanism. and 3) experience apoptosis upon exposure to oxidative stress.
DNA methylation is involved in epigenetic processes such as X-chromosome inactivation, imprinting and silencing of transposons. DNA methylation is a highly plastic and critical component of mammalian development The DNA methyltransferases (Dnmts) are responsible for the generation of genomic methylation patterns, which lead to transcriptional silencing. The maintenance DNA methyltransferase enzyme, Dnmt 1, and the de novo methyltransferase, Dnmt3a and Dnmt3b, are indispensable for development because mice homozygous for the targeted disruption of any of these genes are not viable. The occurrence of DNA methylation is not random, and it can result in gene silencing The mechanisms underlying these processes are poorly understood. It is well established that DNA methylation and histone deacetylation operate along a common mechanistic pathway to repress transcription through the action of methyl-binding domain proteins (MBDs), which are components of, or recruit, histone deacetylase (HDAC) complexes to methylated DNA. As a basis for future studies on the role of the DNA-methyl-transferase in porcine development, we have isolated and characterized a partial cDNA coding for the porcine Dnmt1. Total RNA of testis, lung and ovary was isolated with TRlzol according to the manufacture's specifications. 5 ug of total RNA was reverse transcribed with Super Script II in the presence of porcine Dnmt 1 specific primers. Standard PCRs were performed in a total volume of 50 ul with cDNA as template. Two DNA fragmenets in different position were produced about 700bp, 1500bp and were cloned into pCR II-TOPO according to the manufacture's specification. Assembly of all sequences resulted in a cDNA from 158bp of 5'to 4861bp of 3'compare with the known human maintenance methyltransferase. Now, we are cloning the unknown Dnmt 1 region by 5'-RACE method and expression of Dnmt 1 in tissues from adult porcine animals.
Eleven wrasse species inhabit the coastal waters of Jeju Island, Korea. They are the target of leisure fishing and are considered good eating. We investigated the distribution of standard length (SL) by sex of wrasse in Jeju coastal waters for Halichores poecilopterus, H. tenuispinis, Pseudolabrus japonicus, and Pteragogus flagellifera. A cross-section of the ovary showed the ovarian cavity and ovarian lamellae containing oocytes. A cross-section of the testis showed many lobules containing spermatogonia and spermatocytes. A cross-section of a gonad undergoing sex reversal showed the regression or reduction of oocytes and some spermatocytes located in the ovarian lamellae. A cross-section of a sex-reversed testis showed the primary structure of the ovary, with spermatocytes distributed in the epithelium of the lamellae, and reformed seminiferous ducts in the basement lamellae. (중략)
T. S. 엘리엇의 후기 작품들은 그의 기독교적 세계관, 구원적 시간(역사) 관, 상정시학의 깊은 연관성을 보여주고 있다. 초기의 허무주의적 세계관을 극복한 엘리엇윤 파편화된 현대적 삶은 절대자에 대한 믿음을 복원함으로 써만 치유될 수 있다고 생각한다. 구원적 세계관에 부합하는 시적 언어로 서 엘리엇은 조화, 유기성, 통일성을 담지하고 있는 상정을 중요시 한다. 엘리엇의 상정시학은 감정과 지성, 주관과 객관, 표현과 내용의 통일을 강 조하는 그의 통합된 감수성, 객관적 상관물 이론 둥에 부분적으로 나타나 고 있으며, 또한 단테에서 보들레르에 이르는 형이상학적시 전통을 재해석 하는 1926년 클라크 강연에서 구체화되고 있다. 시간의 측면에서 엘리엇의 상징시학은 순간과 영원의 교차를 중시하며 이 주제는 그의 후기 장시인 「네 사중주』 에서 기존의 직선적 혹은 순환적 시간관에 대한 대안으로서 제시되고 있다. 이 시에서 엘리엇은 현대성의 지배적 역사관인 진보의 서 사와 그 자신의 초기 허무주의 역사관을 지양하고, 동 시대의 영국을 순간 과 영원이 교차하는 시/공간으로 다시 읽음으로써 그의 후기 상징시학과 구원적 역사관의 통합을 보여주고 있다.
This experiment was conducted to investigate the effects of turf grass growth to seedling rates and bed soil types. The results of this experiment were summarized as follows; the more increasing the seedling rates, the plant height and leaf length were longer, but the number of leaf and number of branch were fewer. Incase of transplanting of turf grass seedling by rice transplanting machine, it might be considered that the proper alternative bed soil was sandy loam soil with regarding to the economic aspects. The maximum seedling rate of turf grass in the seedling tray for rice was 1,000 of seedling amount due to the nutrient competition with intensive seedling. As the results indicated, it might be recommended that the proper alternative bed soil was sandy loam soil with 1,000 of seedling rates in case of transplanting of turf grass seedling by rice transplanting machine.
This study was carried out obtain basic information for growth characteristics by different age of Dendropanax morbifera in chonnam, such as Yosu, Sunchon, Haenam, Gangjin, Wando. The pH of soil in cultivation area was 5.2 to 5.6 and organic matter was very high up to 10.6% . P205 content of soil in cultivation area was 35.3 to 42.1 mg/L, C.E.S was 13.9 to 14.4 me/100g, and moisture rate was 19.2 to 21.1 % . The flowering of Dendropanax morbifera began from 6~7 years old tree. The flowering date was at 10th of July at Wando. The growth characteristics of 12 years old tree was 929.5cm in stem height, 134.1mm in stem diameter, and 15 years old tree was 1,117.9cm in stem height, 160.8mm in stem diameter. The number of leaf was 13.9 at five years old tree, and the petioles length was 12.6 at five years old tree. Xylem sap can be had at more than 10 years old tree with good growth more than 10 em stem diameter.
Many ascomycetes were collected at Mt. Minjuji, Mt. Manduck, Mt. Odae, Mt. Yonsuk and Mt. Sunun from August 1999 to September 2000. They were identified. According to the reusulting, Helvella villosa (Hedw.: O. Kuntze) Dissing and Nannfedt, Dasychyphus corticalis (Pers.:Fr.) Mass., Lachnellula fuscosanguinea(Rhem) Dennis, Lachnellula subtilissima (Cke.) Dennis and Scutellina erinaceus (Schw.) Kunrze are newly to Korea. They were designed Korean common names by authors.
In this study, plant regeneration through in vitro culture from plantlet stems of Yooja (C. junos Sieb.) and trifoliate orange (P. trifoliata Rafin.) was attempted to make mass-production system of virus-free plants having the same genotype with mother plant. In order to investigate physiological change depending on the developmental stage of plant regeneration, the changes of total protein, peroxidase and esterase activity and their isozyme patterns as well were examined in 1/2 MS medium. The results are as follows : 1. The MS medium for the optimal callus induction and shoot formation was utilized. The medium was supplemented either with 2,4-D and Kinetin or with BA and NAA. The optimal concentrations were the combination of 1.0mg/ 2,4-D +0.3mg/ Kinetin and 1.0mg BA +0.3mg NAA in callus induction and shoot formation, respectively. 2. For the plant regeneration from somatic embryos, 1/2 MS medium was used with supplements of growth regulators (free, 1.0mg/ IBA +1.0mg/ BA ,0.5mg/ IBA +0.5mg/ BA). Shooting and rooting were the best in the treatment of 0.5mg/ IBA and 0.5mg/ BA combination. 3. The total protein content has a tendency of increase with the developmental stage of embryo, but it was decreased at the plantlet. Also it was the highest at 8 and 6 weeks stage in C. junos Sieb. and P. trioliata Rafin, respectively. In the SDS-PAGE pattern of protein, C. junos Sieb. showed bands of 29.0 and 40kDa at 10 weeks. The 45,66 and 97.4 kDa bands at 10 weeks of culture were shown in P. trifoliata Rafin. 4. The highest esterase activity was shown at the 6 and 8 weeks of culture in C.junos Sieb. and P. trifoliata Rafin.., respectively. 5. Esterase isozyme patterns were shown difference according to the developmental stage. In C. junos Sieb. a new band was observed at pl 7.7 following 4 weeks culture. On the other hand, new bands in P. trifoliata Rafin. were observed at pl 7.5~6.5 following 4 and 6 weeks culture, respectively.