Varieties of Bacillus thuringiensis (Bt) crystal proteins, Cry proteins, have so far been found as one of the most successful biological control agents which are safe to natural environments for a long time. Recently, cry genes encoding these Cry proteins have been widely applied for construction of transgenic crops resistant to pest insects. In this study, through the 3D structure prediction and accompanying mutagenesis study for the Mod-Cry1Ac, 7 and 16 amino acid residues from domain I and II, respectively, responsible for its insecticidal activity against larvae of Spodoptera exigua and Ostrinia furnacalis were identified. To construct novel cry genes with improved insecticidal activity, we randomly mutated these 23 amino acid sequences by in vitro muti site-directed mutagenesis, resulting in totally 24 mutant cry genes. For further characterization, these mutant cry genes were expressed as a fusion protein with polyhedrin using baculovirus expression system. SDS-PAGE analysis of the recombinant polyhedra revealed that expressed Cry proteins was occluded into polyhedra and activated stably to 65 kDa by trypsin. In the further study, we plan to investigate their insecticidal activity against Plutella xylostella, S. exigua and O. furnacalis larvae.

Althogh Spermatogonial stem cells (SSCs) are widely studied in mice, study of pig SSCs is not sufficient for the isolation, long-term culture, and characterization. To identify the effect of growth factor in cultured pig SSC, newly generated pSSCs like cell from neonatal 5days porcine testis were cultured and investigated for the pSSCs like cell formation. Glial derived neurotrophic gactor (GDNF), fibroblast growth factor (FGF), leukemia inhibitory factor (LIF), and epidermal growth factor (EGF) were applied to culture media to identify the pSSC like cell growth and stem cell formation. The criteria for the determining of stem cell characters, morphology, number of colonies, putative stem cell marker were analysed by microspic, polymerase chain reaction (PCR) and immunocytochemistry (ICC) methods. Most of the pSSCs like cells were formed approximately 100 μm size with sphere shape. Most of the feeder cells were highly dependent on FGF that feeder cells were not stably attached on plate without FGF and colony formation of pSSC was not observed consequently. Immunocyto chemistry data revealed that this cells expressed the ubiquitin-C-terminal hydrolase 1 (UCHL-1, PGP9.5) and Dolichos Biflorus Agglutinin (DBA) in addition of 20 ng/ml EGF, 10 ng/ml FGF, 10 ng/ml GDNF, 10 U3/ml LIF. In addition, Alkaline Phosphatase ()was positive in all period of culture. This study suggest that various growth factorsinp SSC culture system is important to regulate and maintain the pSSC. In conculsion, although the precise role of growth factor in pSSC proliferation need to be clarified, combination of growth factor might be critical in order to derivation and proliferation of neonatal pSSCs and spermatogenesis.

Spermatogonial stem cells (SSC) undergo self-renewal division and generate spermatogenesis to produce mature spermatozoa. Very recently in some species, include rodent and farm animals, SSC has been isolated and cultured in vitro. In this study, we analysed SSC marker of both 5 days old and pubertal pig testis by histological method. In 5day pig testis, prior to set of spermatogenic differentiation, the seminiferous tubules contain a relatively large number of SSCs than in pubertal pig testis. Then putative pig SSCs were successfully isolated from 5 day pig testis, and cultured long term using CD34 positive cell culture media contained GDNF, bFGF, LIF and EGF. The SSC colonies were appeared at 3 days after cells were seed. The SSC colonies were alkaline phosphatase positive and strongly expressed PGP 9.5, PLZF and DBA, but not expressed GATA4 and LH receptors. The SSC colonies were stably proliferated in GDNF, bFGF, LIF and EGF contained CD34 positive cell culture media up to 60 days. This study will be facilitated to study of in vitro and ex vivo spermatogenesis and of production of transgenic pigs using sperm vector.

INTRODUCTION In rodent, molecular markers of spermatogonia, spermatocyte, spermatid and sperm have been identified. It has been reported that DBA, PGP 9.5 and NanpG can be the markers for spermatogonia in pig. For further understanding the spermatogenesis of the pig on morphological and molecular level, we report identification of testicular cells in neonatal and pubertal pig testis, and a putative marker for spermatogonia and spermatid in pig testis. MATERIALS AND METHODS Neonatal (3 day) and pubertal testis (150 day) was cut and fixed in Bouin’s solution for immunohistochemistry (IHC). Gonocytes were isolated from neonatal testis for immunocytochemistry (ICC). Based on references (Phillips et al., 2010), thirteen antibodies (VASA, Oct4, NanoG, PGP 9.5, DAZL, SCF, GFR-alpha 1, PLZF, c-kit, integrin-beta1, Thy1, Sohlh1 and CD9) were used for IHC and ICC. Paraffin section was performed for IHC. Gonocytes were attached to the APS-coated slides for ICC. HRP-conjugated and florescent-labeled secondary antibody was used for IHC and ICC, respectively. RESULTS In histological analysis, spermatogonia and Sertoli cells, which are enclosed by seminiferous tubule and connective tissue, were observed in neonatal testis. However, complete spermiogenesis, including spermatocyte, spermatid and spermatozoa, was not observed in neonatal testis. Spermatocyte, spermatid and elongated spermatid were observed in pubertal testis but matured spermatozoa were not observed. As a result, two antibodies (PGP 9.5 and GFR-alpha1) of thirteen antibodies were available for IHC and ICC. As reported in other studies, PGP 9.5 protein was detected in spermatogonia of ne-onatal in IHC. In addition, it was observed in spermatogonia of pubertal testis. GFR- alpha1 protein was detected in spermatogonia of neonatal testis and spermatid of pubertal testis. In ICC, PGP 9.5 protein was detected in gonocytes as reported in other studies. GFR-alpha1 was also observed in gonocyte isolated from pig testis. In this study, we found that 1) only spermatogonia exist in neonatal pig testis (3 day), 2) GFR-alpha1 is a new marker for spermatogenesis in pig testis.

Interferon induced transmembrane protein-1 (IFITM1) is one of transmembrane protein which is differentially expressed in uterus during estrus cycle and pregnancy, that IFITM1 gene is highly expressed in estrus stage by the effect of estrogen, and in parturition by the effect of PGF2 alpha. This genes are also up-regulated in cells with hyperactivation of the WNT/β-catenin signaling pathway. In this study, to identify a function of IFITM1, the binding partner of IFITM1 were determined using immunoprecipitation and LC- MASSMASS methods. 1, 3 and 5 ug of polyclonal anti-IFITM1 antisera were used for immunoprecipitation, and the 75 kDa of specific band was detected in silver stained polyacylamide gel. This band were chracterized using LC-MASS-MASS, and revealed this band is glucose regulated protein 75 (GRP75) which binds to p53 and inhibits the p53 action in nucleus. To identify the localization of GRP75 in cells, immunocytochemical approach has been applied, and GRP75 is expressed in mitochondria of L929 murine connective tissue cells. Co-localization study between IFITM1 and GRP75 in L929 cell identified that these two proteins were closely expressed in mitochondria. Although the role of the interaction of these two protein need to be clarified in various biological phenomena, this data suggest that close interaction of IFITM1 and GRP75 may regulate cellular functions in uterus on sets of estrus cycle and pregnancy.

SERPINB3 (also known Squamous cell carcinoma antigen 1, SCCA1) is involved in apoptosis, immune response, cell migration and invasiveness of cells. It has been investigated in various types of squamous cell carcinoma. Therefore we investigated the functional role of SERPINB3 gene in human epithelial ovarian cancer (EOC) using laying hens, the most relevant animal model. In 136 laying hens, EOC was found in 10 (7.4%). We compared the expression and localization of SERPINB3 using RT-PCR, quantitative RT-PCR, in situ hybridization and immunohistochemistry, and SERPINB3 activation was detected in chicken and human ovarian cancer cell lines using immunofluorescence microscopy. Thereafter, we examined the prognostic value of SERPINB3 expression in patients with EOC by multivariate linear logistic regression and Cox’ proportional hazard analyses. In present study, SERPINB3 mRNA was induced in cancerous ovaries (p< 0.01), and it was only expressed in the glandular epithelium(GE) of cancerous ovaries of laying hens. SERPINB3 protein was localized predominantly to the nucleus of glandular epithelium in cancerous ovaries of laying hens, and it was abundant in the nucleus of both chicken and human ovarian cancer cell lines. In 109 human patients with EOC, 15 (13.8%), 66 (60.6%) and 28 (25.7%) of those patients showed weak, moderate and strong expression of SERPINB3 protein, respectively. Strong expression of SERPINB3 protein was a prognostic factor for platinum resistance (adjusted OR, 5.94; 95% Confidence Limits, 1.21-29.15). Therefore SERPINB3 may play an important role in ovarian carcinogenesis and be a novel biomarker for predicting platinum resistance and a poor prognosis for survival in patients with EOC. This research was funded by the World Class University (WCU) program (R31-10056), Basic Science Research Program (2010- 0013078) through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science, and Technology and by the Next-Generation BioGreen 21 Program (No.PJ008142), Rural Development Administration, Republic of Korea.

A new cultivar "KNR11-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2598 and KNR2610. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR11-1" was one day faster than that of control strain KNR2312. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has the teeth of a comb. The yield was 90.4 g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. It is able to store up to 67.5 days in a cold room. Analysis of the genetic characteristics of the new commercial strain "KNR11-1" showed a different profile as that of the control strain KNR2312 when Random Amplified Polymorphic DNA primers were used. The new cultivar of Pleurotus eryngii is characterized by the improved storability after harvesting.

The occurrence and rapid range expansion of a fulgorid, Lycorma delicatula(White), has recently been reported in korea. It was previously known to occur in China, Japan, Vietnam and India. The first occurrence fo L. delicatula was reported in 2004 in the Cheonan. It has one generation per year and overwinters as egg masses on the bark of host trees. The number of egg per egg mass was 32.69±6.49. The cumulative ratio of hatchability at different temperature was the highest at 25℃ and the lowest at 10, 30℃. The egg hatchability was investigated in different regions in Chung-buk. Since L. delicatula eggs has been stored in -20±1, 0±1, 5±1℃ conditions for 1 ∼60 days, the cumulative ratio of hatchability was 2% stored in -20±1℃ for 3days, But hatchability was 52, 48% stored in 0±1℃ for 3, 14days and 82, 68% stored in 5±1℃ for 3, 14days. And hatchability was 0% for 60days. By straight regression equation(the growth ratio/treatment degree) the growth zero degree of L. delicatula was 10.4℃. The hatchability of L. delicatula in Cheongwon-gun, Okcheon-gun, Cheongju-city where’s winter lowest temperature was over -19℃ was over 79%. But The hatchability of L. delicatula in Jincheon-gun where’s winter lowest temperature was less than -19℃(continuos two days) was 35%.

The notion that dental amalgam is a potential source of mercury exposure remains a controversial issue. However, there are few epidemiological analyses that have addressed whether this occurs in children. We aimed in our current study to identify the relationship between dental amalgam filling surfaces and the blood mercury levels in a cohort of 711 South Korean children aged between 8-9 years. Oral examinations were conducted to detect the number of amalgam filling surfaces on the teeth of these individuals. Blood samples were also taken from these children to assess the levels of mercury accumulation in the body. The amalgam filling surfaces were classified into four groups based on their number: 0 (n = 368), 1-5 (n = 219), 6-10 (n = 89), and 11+ (n = 35). The blood mercury levels in the children with more than 10 amalgam surfaces was 0.47 μg/L higher on average than those with no amalgam surfaces after adjusting for the frequency of fish or seafood consumption, age, and gender (P < 0.05). We found from our data that a higher number of dental amalgam fillings correlated with a higher blood mercury level in Korean children. Further studies are needed to investigate whether these elevated mercury levels exert neurotoxic or nephrotoxic effects.

A new cultivar "KNR10-1" of Pleurotus eryngii was developed by the method of mono-mono crossing between monokaryotic strains derived from KNR2594 and KNR2610, respectively. The optimum temperature of mycelial growth was 25℃ and that of fruiting body development was 15 - 16℃. The period of harvesting including primordia formation of "KNR10-1" was one day faster than that of control strain KNR2312. The color of pileus and stipe surface was neutral-brown and pure white, respectively. The shape of pileus was dome and has the teeth of a comb. The yield was 90±12.9g/850cc plastic bottle. The fruit body has a longer shelf life than control strain KNR2312 at 4℃. Analysis of the genetic characteristics of the new commercial strain "KNR10-1" showed a different profile as that of the control strain KNR2312 when RAPD (Random Amplified Polymorphic DNA) primers were used. This new variety of King Oyster mushroom is characterized by the improved storability after harvesting.

According to the IPCC report (IPCC, 2001), the global-mean surface temperature has risen by 0.6 oC during the 20th century due to the increase of atmospheric concentrations of greenhouse gases, and the mean temperature of the Korean Peninsula is also risen by 1.5 oC during the same period due to global warming as well as rapid urbanization. During the 21st century, the global-mean temperature is projected to rise 1.5-5.8oC associated with the various scenarios from IPCC Special Report on Emission Scenario (SRES) (2000). Climate change is expected to have important impacts on the relationship between crops and insect pests as well as on human societies. Frazier et al. (2006) demonstrated that warm-adapted insect species have much higher maximum population growth rate (intrinsic rate of increase) than do cold-adapted species; which indicates biochemical and physiological adaptations of insects do not overcome the constrain of thermodynamics. Consequently, global warming could lead to an increase in the number of insects worldwide. This presentation will discuss the changes in the population abundance of several citrus pests according to global warming. The differences of population growth rates between normal year's or past temperatures and elevated temperatures were compared, and also analytical models such as matrix model and predator-prey model were applied to project the performance of population dynamics of some pests and natural enemy.

Deciduous tree fruits such as pears and apples are widely grown in the Pacific Northwest of the United States and are well adapted to the seasonal environment in that region. Extended cold periods provide adequate chilling to break dormancy and reinitiate growth in the spring. Cold exposure synchronizes the physiological processes and makes sure that bloom is uniform and that fruit matures in a uniform manner. As a result of global warming, some fruit-growing areas may experience inadequate cold exposure during the winter months, gradually shifting the southern boundary for production of deciduous fruits further north. However, climate change will affect not only growth and fruiting habits of fruit trees but also the insect and mite pests which feed on them. There is general agreement that in temperate regions a trend towards warmer summers and milder winters will generally benefit insect and mite pests and increase their injuriousness. Temperature changes in particular will impact the development, mortality, phenology, and voltinism of fruit pests. Here we discuss how climate change may affect pests and control practices on deciduous tree fruits in the Hood River Valley of northern Oregon. This small valley extends in a north-south direction from the Columbia River to the foothills of Mt. Hood and is characterized by a varied topography and large altitudinal differences (sea level to 600 m). The major pest of pears and apples in that area is codling moth, Cydia pomonella L., a cosmopolitan pest which is present in most deciduous fruit-growing areas of the world. Like its host trees, the codling moth is well adapted to a seasonal environment. Diapause is the principal mechanism which synchronizes its phenology with the tree and the presence of fruit, the larval food source. Diapausing overwintering larvae require cold exposure (chilling) to terminate diapause in late winter or early spring. At the lower elevations close to the Columbia River the codling moth is bivoltine but gradually becomes univoltine at the higher elevations where the growing season is shorter and fewer heat units (above 10oC) are available for development. Long-term temperature records from the lower Hood River Valley indicate that the 25 years since 1985 have been considerable warmer than the 25 years prior to 1985. For instance, the average heat units available for codling moth development over a season have increased by more than 10% over the last 25 years. The codling moth is adapting to this warming trend by gradually increasing its voltinism (number of generations). As a result, the severity of codling moth as a pest can be expected to increase. Therefore, fruit growers will have to adjust and intensify control practices to keep fruit free from codling moth damage. We will also explore how other fruit and foliage feeding pests which are part of the pest complex of pears and apples in northern Oregon fare under different global warming scenarios compared to codling moth.