Soluble-NSF attachment protein receptor (SNARE) proteins play a role in vesicle fusion, exocytosis, and intracellular trafficking in neuronal cells as well as in fertilization and embryogenesis. We investigated the expression patterns of two SNARE proteins, SNAP-25 and synaptotagmin VII (SytVII), and their regulation by pregnant mare serum gonadotropin (PMSG) during mouse ovarian follicular development. Ovaries were obtained at 0, 12, 24, 36, and 48 h post-PMSG injection of immature mice. SNAP-25 and SytVII mRNA expression levels increased gradually in a time-dependant manner. However, protein levels revealed different patterns of expression, suggesting different translational regulation following PMSG stimulation. SNAP-25 and SytVII expression was closely associated with thickening of the granulosa cell (GC) layer and follicle morphological changes from a flattened to a cuboidal shape. To explore follicle stimulating hormone receptor (FSHR)-mediated regulation of their expression, GCs from preantral follicles were cultured to examine the effects of FSHR siRNA knockdown. FSHR siRNA abolished upregulation of the SNAREs in both PMSG and FSH-stimulated GCs. This abolished gene expression was rescued by adding dibutyryl cyclic AMP to the cultures. These results suggest that SNAP-25 and SytVII expression is regulated via the FSHR-cAMP pathway during follicular development.

Construction of the Asian Network for Sustainable Organic Farming Technology (ANSOFT) will be cooperatively administered by the public researchers in 12 Asian member countries from 2010 to 2012. ANSOFT will bring forward multiple reports, which will be constantly renewed by the member countries, regarding environmental issues, plant and landscape protection techniques, regulations and policies of each country’s government on an organic ag끼culture， and natural resources such as organic seeds and biological agents.

Controllable transgenic expression systems in transgenic animal model are valuable to the development of therapeutic approaches in human medical fields. The aim of this study was to 1) produce a transgenic cloned dog using inducible tetracycline vector system, and 2) investigate whether the transgenic cloned dog could be induced the transgene expression using doxycycline (Doxy). Canine fetal fibroblasts were infected with retroviral vectors designed to express the enhanced green fluorescent protein (eGFP) gene under the control of tetracycline-inducible promoter. For somatic cell nuclear transfer (SCNT), nucleus of an in vivo matured oocyte was removed and an eGFP expressed cell cultured with 1 ㎍/㎖ of Doxy was injected. After electrical fusion and chemical activation, the reconstructed embryos were transferred to a recipient and pregnancy diagnosis was performed by ultrasonography. Experiment I evaluated the mean fluorescence intensity (MFI) of infected cells while the cells were cultured in the presence of 1 ㎍/㎖ of Doxy for 5 days, and then in the absence of Doxy for 7 days using fluorescence-activated cell sorter. Experiment II was designed to produce an eGFP controllable transgenic cloned dog via SCNT. For verification of transgenic dog, experiment III was performed Southern Blot analysis and observation in vivo regulation of eGFP expression in the cloned dog treated with 100 ㎎/㎏ of Doxy every 2 days for 2 weeks under ultraviolet light. In experiment IV, western blot was used to detect eGFP increase and decrease in skin tissues of transgenic dog under the presence or absence of Doxy. In the results of experiment I, the MFI for infected cells was rapidly increased to approximately 42.3 times after 3 day-treatment compared to pre-treatment and quickly decreased 3 days after ceasing the treatment. In experiment II, a total of 203 embryos were transferred to nine recipients and three pregnant delivered three pups (Tet-on eGFP 0, Tet-on eGFP 1, and Tet-on eGFP 2) by C-sec and Tet-on eGFP 2 among them is still alive. All cloned pups were genetically identical to the donor cell. Tet-on eGFP 2 showed an apparent in vivo eGFP expression on her body after Doxy administration in experiment III. The result of Sothern blotting showed that the transgene insertion was detected from the three cloned dogs and all organs of Tet-on eGFP 1. Experiment IV indicated that a robust eGFP expression in skin tissue of Tet-on eGFP 2 rapidly increased after Doxy treatment and gradually decreased to basal level on 9 weeks after ceasing the treatment. In conclusion, we report here for the first time an inducible transgenic system in canine species and it can stably induce the transgene expression at intended time. This study has demonstrated the capacity to generate transgenic model dog which could regulate the transgene and it would contribute to human medical research fields.

"Early Valley", is an early maturing potato cultivar with high yield potential. "Early Valley" is a clonal selection resulting from the cross between 'Suncrisp' and 'A87109-10'. It has medium plant height and light green foliage. "Early Valley" has medium flowering habit and white flowers. Tubers are smooth, yellow skin, light yellow flesh, round tuber shape, medium eye depth, and medium dormancy and good keeping quality. It has stable yield under wide range of climatic conditions. "Early Valley" is resistance to late blight, but moderately susceptible to common scab and hollow heart. This cultivar is also resistant to potato rotting at harvesting during the raining season. "Early Valley" has high level of antioxidant activity (about three times higher) and vitamin C (higher by 40%) than the 'Superior'. This cultivar has high level of tuber uniformity and capable of yielding 36.56 t/ha which is 17.07% higher than the control potato cultivar 'Superior' under optimum agronomical practices.

The purpose of this is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine embryos. Blastocysts were produced by in vitro fertilization of slaughterhouse-derived, in vitro matured oocytes with frozen-thawed boar semen, and subsequent culture on granulosa cell monolayer. After frozen-thawing, embryos were culture in NCSU-23 medium with 5 mM hypotaurine, 4 mg/ BSA and 10 ng/ for 48 hrs to survival tests. When blastocysts were frozen-thawed by OPS methods, the embryos with normal morphology were 32.1, 34.5 and 38.9 % in early blastocyst, blastocyst and expanded blastocyat stages. The rates of partial damaged embryos were significantly (P<0.05) higher in early biastocysts than expanded blastocysts. In another experiment, the embryos frozen by OPS methods were cultured for 48 hrs for survival and developmental rates in vitro. The proportions of embryos hatched were 11.8, 20.2 and 33.3% in embryos frozen-thawed at stages of early blastocyst, blastocyst and expanded embryos. On the other hand, The proportions of embryo with normal morphology after culture were 23.5, 25.0 and 33.3% in embryos frozen-thawed at stages of early blastocyst, blastocyst and expanded embryos. These finding indicate the possible broader application for OPS methods that this procedure described is relatively harmless, that it can be used for blastocysts of different developmental stages.

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.

It was found that the purified extract from A. gigas Nakai (polysaccharide, M.W., 25 kD) controled differentiating human ES cells. Its optimal supplementation concentration was decided as 0.8 (μg/ml) to efficiently control the differentiation. It also enhanced the cell growth, compared to the control. However, most widely used and commercially available differentiating agent, Leukemia Inhibitory Factor (LIF) negatively affected on the cell growth even though it controls the differentiation of ES cells, down to 40-50 % based on morphological observation and telomerase activity. It was presumed that the extract first affected on cell membrane and resulted in controlling signal system, then amplify gene expression of telomere, which enhanced the telomerase activity up to three times compared to the control. LIF only increased the enzyme activity up to two times. It was confirmed that the extract from A. gigas Nakai could be used for substituting currently used differentiation controlling agent, LIF from animal resources as a cheap plant resource and not affecting the cell growth. It can broaden the application of the plants not only to functional foods and their substitutes but also to fine chemicals and most cutting-edge biopharmaceutical medicine.

This study is being done for the purpose to achieve a stable demand and supply of materials for long term. The author investigated condition of suitable mixer design method for concrete, an individual value and a character of dynamics as a supplement material by drying up the natural aggregate, and a possible practicalization plan to reduce defects to the minimum. As a result of this study, it can be acquired a good quality of concrete by the use of the natural sand (Nak Dong river sand) and the crushed sand with suitable mixing ratio. The fineness modulus (F.M) of mixing aggregate is 2.7±1 at this time and this is good for workbility and plasticity of concrete.

This work aims to : establish a model of the container physical distribution system of Pusan port comprising 4 sub-systems of a navigational system, on-dock cargo handling/transfer/storage system, off-dock CY system and an in-land transport system : examine the system regarding the cargo handling capability of the port and analyse the cost of the physical distribution system. The overall findings are as follows : Firstly in the navigational system, average tonnage of the ships visiting the Busan container terminal was 33,055 GRT in 1990. The distribution of the arrival intervals of the ships' arriving at BCTOC was exponential distribution of Y=e-x/5.52 with 95% confidence, whereas that of the ships service time was Erlangian distribution(K=4) with 95% confidence, Ships' arrival and service pattern at the terminal, therefore, was Poisson Input Erlangian Service, and ships' average waiting times was 28.55 hours In this case 8berths were required for the arriving ships to wait less than one hour. Secondly an annual container through put that can be handled by the 9cranes at the terminal was found to be 683,000 TEU in case ships waiting time is one hour and 806,000 TEU in case ships waiting is 2 hours in-port transfer capability was 913,000 TEU when berth occupancy rate(9) was 0.5. This means that there was heavy congestion in the port when considering the fact that a total amount of 1,300,000 TEU was handled in the terminal in 1990. Thirdly when the cost of port congestion was not considered optimum cargo volume to be handled by a ship at a time was 235.7 VAN. When the ships' waiting time was set at 1 hour, optimum annual cargo handling capacity at the terminal was calculated to be 386,070 VAN(609,990 TEU), whereas when the ships' waiting time was set at 2 hours, it was calculated to be 467,738 VAN(739,027 TEU). Fourthly, when the cost of port congestion was considered optimum cargo volume to be handled by a ship at a time was 314.5 VAN. When the ships' waiting time was set at I hour optimum annual cargo handling capacity at the terminal was calculated to be 388.416(613.697 TEU), whereas when the ships' waiting time was set 2 hours, it was calculated to be 462,381 VAN(730,562 TEU).