The ability of plants to endure environmental stress factors, which are going to be more severe due to global warming, is important especially for forest plants. Because obtain trait of resistance to temperature using conventional breeding for woody plants is a time consuming way. In this study, chloroplast-localized OsHSP26 gene was overexpressed in Populus alba L. to breed tolerant transgenic poplar to temperature stress. The plantlets of OsHSP26-overexpressed transgenic poplar showed more heathy phenotypic response than wild-type plants under both prolonged low- and high-temperature stress. While the SPAD value, which refers chlorophyll content, in wild-type plants decreased depending on the exposure time to the temperature stress, higher SPAD value were shown in the transgenic plants. The contents of total phenolic compounds in the transgenic plants were lower than those of the wild-type plants, and not significantly changed except in the treatment of prolonged low-temperature. However, the total flavonoids contents of the transgenic plants were dramatically increased under prolonged temperature stress. The DPPH scavenging activities of the transgenic plants were higher than those of the wild-type plants under temperature stress. Consequently, it was revealed that overexpressing OsHSP26 allow for P. alba to be tolerant to temperature stress.

Terfenadine (TFN) was a second generation histamine receptor antagonist. Although several studies have reported the regulatory effect of H1-histamine receptor antagonists in human cancer cell lines, its effect in oral cancer remains unclear. In this study, we focused on addressing the anti-cancer activity of TFN in human oral cancer cell lines. The anti-cancer activities of TFN were performed by tryphan blue exclusion assay, 4'-6-diamidino-2-phenylindole (DAPI) staining, live/dead assay and Western blot analysis. TFN induced a significant reduction of the growth in three different human oral cancer cell lines (MC3, HSC4 and Ca9.22). TFN markedly induced apoptosis through DNA damage and increase in cytotoxicity. It also accumulated cleaved PARP and caspase 3. This process was due to cleavage of caspase 8 and Bid protein. The results from this study strongly demonstrated that the cleavages of caspase 8 and Bid are required for the apoptotic activity of TFN in human oral cancer cells. Taken together, these findings suggest TFN as a potent anticancer drug candidate for the treatment of oral cancer.

Phasianus colchicus is not only a beautiful bird but also a great value in science and under the threat of endanger. Hence, the aim of this study was to isolate the pheasant male germ cells (mGCs) and then induce them into elongated sperm-like cells in vitro. The mGCs were purified and enriched by a two-step plating method based on the different adherence velocities of mGCs and somatic cells. The percentage of the c-kit positive cells and c-kit negative cells examined by flow cytometry analysis (FCA) was 92.87% and 2.57%, respectively. Subsequently, the mGCs were induced for 48h in DMEM/F12 medium supplemented factors such as retinol acid, testosterone and bovine FSH, followed by 5 weeks in culture. We found that some elongated sperm-like cells appeared initially in vitro under inducement of stimulated factors. The elongated sperm-like cells showed in the expression of changed morphology and post-transcriptional marker such as spermatid associated (SPERT), spermatid perinuclear RNA binding protein (STRBP), round spermatid basic protein 1 (RSBN1) and SPER1L. Moreover, in DNA content identified assay, induced cells showed that the 1C DNA population markedly increased in differentiated group but it was not change in undifferentiated group. Successful in vitro differentiation of pheasant testicular germline cells into spermatids appears to offer extremely attractive potential for the conservation of endangered birds and treatment of male infertility.

Nutritive value analysis of feed is very important for the growth of livestock, and ensures the efficiency of feeds as well as economic status. However, general laboratory analyses require considerable time and high cost. Near-infrared reflectance spectroscopy (NIRS) is a spectroscopic technique used to analyze the nutritive values of seeds. It is very effective and less costly than the conventional method. The sample used in this study was a corn kernel and the partial least square regression method was used for evaluating nutrient composition, digestibility, and energy value based on the calibration equation. The evaluation methods employed were the coefficient of determination (R2) and the root mean squared error of prediction (RMSEP). The results showed the moisture content (R2val=0.97, RMSEP=0.109), crude protein content (R2val=0.94, RMSEP=0.212), neutral detergent fiber content (R2val=0.96, RMSEP=0.763), acid detergent fiber content (R2val=0.96, RMSEP=0.142), gross energy (R2val=0.82, RMSEP=23.249), in vitro dry matter digestibility (R2val=0.68, RMSEP=1.69), and metabolizable energy (approximately R2val >0.80). This study confirmed that the nutritive components of corn kernels can be predicted using near-infrared reflectance spectroscopy.

The objectives of this study were to evaluate the antimicrobial effects of ε-poly-L-lysine mixture (EPM) on cheon-ggukjang and to determine the optimal mixing ratio in cheonggukjang. Increasing the EPM up to 2% decreased via-ble cell counts at both 15oC and 30oC storage. The pH of cheonggukjang without EPM increased slowly over thestorage periods, but cheonggukjang with EPM showed a slight increasing tendency, with the highest pH at 2% EPM.The sensory scores, such as texture and flavor, were highest in samples containing 2% EPM; however, overall pref-erence was not significantly different when compared to the control. For the optimum cheonggukjang processingconditions, 13 experimental points were selected. Soybean and EPM were chosen as independent variables. Viablecell count, pH, texture, and overall preference were measured. The optimum formulation of cheonggukjang usingthe numerical analysis was found to be 98.52% soybean and 1.48% EPM, resulting in a 0.722 desirability value.

In the present study, lichen(Parmelia sp.) extract showed insecticidal activity against Aedes albopictus, and the effect of growth inhibiting activity was investigated. Acetone and methanol extracts of the lichen against larvae of Aedes albopictus were showed high insecticidal activities in low lethal concentration. 50% lethal concentration of the acetone extractof the lichen is 0.13% and the 50% lethal concentration of methanol extract of lichen is 0.15% respectively. This experiment that used acetone and methanol extracts of the lichen were observed for 24, 48 and 72 hours. As a result, the higher concentration and the longer exposure time is increased mortality against Aedes albopictus. Pupation time took more time as the higher concentration of acetone extract of lichen. Consequently, the lichen extract is effect in inhibiting the growth of Aedes albopictus larvae. In this experiment indicates that lichen extract has activity against Aedes albopictus and is available as the natural insecticide.

Recently, several approaches have been used to measure the oxygen consumption rates of individual embryos, but relationship between oxygen consumption and pregnancy rates of Hanwoo following embryo transfer has not yet been reported. In this study, we investigated the correlation between oxygen consumption rate and pregnancy rates of Hanwoo embryo using a SECM. In addition to, the expression of apoptosis-related genes was determined using real-time PCR by extracting RNA according to the oxygen consumption of in vivo embryo. First, we found that the oxygen consumption significantly increased in blastocyst-stage embryos (blastocyst) compared to early blastocyst stage embryos, indicating that oxygen consumption reflects the embryo quality (Grade I). The oxygen consumption or GI blastocysts were significantly higher than those of GII blastocysts (10.2x1014/mol s-1 versus 6.4x1014/mol s-1, p＜0.05). Pregnant rate in recipient cow was 0, 60 and 80% in the transplantation of embryo with the oxygen consumption of below 10.0, 10.0~12.0 and over 12.0x1014/mol s-1, respectively. Apoptosis regulatory genes, Hsp-70.1 were significantly increased in over-10.0 group than below 10.0 group but in Caspase-3, Bax and P53 gene, there was no significant difference. In conclusion, These results suggest that measurement of oxygen consumption maybe help increase the pregnant rate of Hanwoo embryos.

Using the spray pyrolysis process, nano-sized cobalt oxide powder with average particle size below 50 nm was prepared from cobalt chloride solution. The influences of the raw material solution on the properties of the powder formed examined. When the concentration of Co was low(20 g/L), the average particle size of the powder formed was roughly 20 nm, and the cohesion between these particles was significantly strong. When the concentration of Co increased to 100 g/L, the droplets nearly failed to exist in circular form and reflected a severely divided form. Furthermore, the average size of the particles formed was roughly 40 nm, and the particles reflected a polygonal form. When the solution was increased to nearly saturation level (Co at 200 g/L), the particle size distribution reflected significant unevenness due to severe droplet division while the surface also reflected significant unevenness. Furthermore, the average size of the particles formed increased significantly to 70 nm. The results of XRD analysis showed that the strength of the peaks reflected very little change when the concentration of Co was increased from 20 g/L to 50 g/L. Alternatively, when the concentration was increased to 100 g/L, the strength of the peaks increased compared to when the concentration was 50 g/L. However, when the concentration was increased to 200 g/L, the strength of the peaks failed to reflect significant change compared to when the concentration was 100 g/L. The specific surface area dramatically decreased by 30 % when the concentration of Co was increased from 20 g/L to 50 g/L. Alternatively, when the concentration of Co the solution increased to 100 g/L, the specific surface area decreased by roughly 15 %. Furthermore, when the concentration of Co was increased to nearly saturation level(200 g/L), the specific surface area decreased by roughly 35%.

Polyhedrin is the major component of the nuclear viral occlusions produced during replication of the baculovirus Autographa californica multicapsid nucleopolyhedrovirus (AcMNPV). To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of various polyhedrin fragments were investigated by fusion expressing them with the enhanced green fluorescent protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the previously reported minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The marked increase of EGFP by these fusion expressions was confirmed through protein and fluorescence intensity analyses. Among the fusion-expressed protein in nucleus and cytoplasm, the most hyper-expression was observed in the fusion of amino acids 19 to 110 and 32 to 59 of polyhedrin. Also these fragments, some degradation of only the fused polyhedrin was observed in the fusion of amino acids 19 to 85 and 32 to 85. The production of E2 protein, which is a major antigen of classical swine fever virus, was dramatically increased by fusion expression with polyhedrin amino acids 19 to 110, and its preliminary immunogenicity was verified using experimental guinea pigs. The production of luciferase was approximately two folds increased by fusion expression with polyhedrin amino acids 32 to 59, and its activity was measured using Luminometer. This study suggests a new option for higher expression of useful foreign recombinant protein using the partial polyhedrin fusion expression in baculovirus.

Entomopathogenic fungi are natural pathogens of insects and contribute to the regulation of host insect populations in the environment. Several these fungi produce a wide range of secreted enzymes, secreted protein toxins and secondary metabolites to overcome host defenses and ultimately kill the host, and to defend host resources against competing pathogens and saprophytes. Therefore, this study was performed to select the antimicrobial activity of entomopathogenic fungi form Korea soils against plant pathogenic bacteria Ralstonia solanacearum and plant pathogenic fungi Botrytis cinerea using dual culture technique on SDYA. In addition, we also performed to screening of 2,2-diphenyl-1-picrylhydrazyl (DPPH) radicals scavenging activity compounds from liquid culture filtrates of entomopathogenic fungi and investigate to it’s anticancer activity. As results, 12 isolates, 6 isolates and 25 isolates showing of these fungal metabolites produced antibacterial, antifungal and radicals scavenging activity compounds, respectively. The preferential antimicrobial and radical scavenging activities give evidence that these entomopathogenic fungal metabolites might be useful as a source for plant pathogen control and pharmaceutical interests.

The bulb mite (Rhizoglyphus echinopus) damages garlic, shallot and onion in the bulbs, corms and tubers. It has recently become a serious problem because of the continuous use of acaricides resulting in resistance among bulb mite population. Thus, there is need to find alternative control measures to suppress bulb mite population. Here, we report the screening result of pathogenic fungi for the control of R. echinopus. Initial screenings were performed using 352 isolates of entomopathogenic fungi from Korea soils. As results, 15 isolates of acaropathogenic fungi showed the pathogenicity to bulb mite supporting fungal conidiation. These isolates were identified as 3 isolates of Metarhizium flavoviride var. pemphigi and 12 isolates of Metarhizium pingshaense by microscopic examination and genetic sequencing of the ITS region and elongation factor-1 alpha. Selected 15 isolates were tested for their virulence against adult R. echinopus and the thermotolerance and the activity to UV-B irradiation of conidia. Additionally, the activities of chitinases and proteases produced by M. pingshaense were compared according to the medium. These acaropathogenic fungi would be considered promising for biological control of bulb mite.

Entomopathogenic fungi are natural pathogens of insects and contribute to the regulation of host insect populations in the environment. Several these fungi produce a wide range of secreted enzymes, secreted protein toxins and secondary metabolites to overcome host defenses and ultimately kill the host, and to defend host resources against competing pathogens and saprophytes. This study was performed to evaluate the antimicrobial activity of 207 entomopathogenic fungi form Korea soils against plant pathogenic bacteria Ralstonia solanacearum and plant pathogenic fungi Botrytis cinerea using dual culture technique on SDYA. As results, twelve isolates (5.7%) and six isolates (2.8%) showing the greatest inhibition against R. solanacearum and B. cinerea, respectively. The culture supernatant of these selected isolates completely suppressed the growth of the pathogen, indicating that suppression was due to the presence of antimicrobial compound in the culture filtrate. The stability test of the culture filtrate showed that the antimicrobial component was heat stable and not protein. These entomopathogenic fungal metabolites may be a good feature to be used in the development of a new biocontrol method of R. solanacearum and B. cinerea.

To enhance the production efficiency of foreign protein in baculovirus expression system, the effects of polyhedrin fragments were investigated by fusion expression them with the enhanced green fluorescence protein (EGFP). Recombinant viruses were generated to express EGFP fused with polyhedrin fragments based on the minimal region for self-assembly and the KRKK nuclear localization signal (NLS). The increase of EGFP production by fusion expressions was confirmed through protein and fluorescence intensity analyses. The importance of nuclear localization for enhanced production of EGFP was shown by the mutation of the NLS within the fused polyhedrin fragment. Among the fusion expressed protein in cytoplasm, the most hyper-expression was observed in the fusion of amino acids 32 to 59 of polyhedrin. Polyhedrin fragment fusion expression with classical swine fever virus E2 protein also resulted hyper-enhanced expression of E2 protein. However, the fusion expression of porcine circovirus ORF2 with polyhedrin fragment did not show significant enhance of ORF2 production. These results suggested that the enhancement of foreign protein production when fused with polyhedrin is caused by the enhanced stability of expressed protein.

Hyphantria cunea is a fall webworm is considered an agricultural pest. It is a major pest of many board-leaved trees. H. cunea nucleopolyhedrovirus (HcNPV) and H. cunea granulovirus (HcGV) were isolated from the fall webworm cadavers in Korea. To better understand HcNPV and HcGV, their genomic sequences were determined, analyzed and compared to two viruses together. The entire nucleotide sequence of the HcNPV genome was fully sequenced using 454 pyrosequencing. The genome of the HcNPV was 131,302 bp with a 45 % G+C content. Computer assisted analysis predicted 146 open reading frames (ORFs) of 50 or more amino acids that showed minimal overlap. Further more, when the phylogenetic relationship was analyzed, HcNPV was closely related to Orgyia pseudotsugata MNPV (OpMNPV) which belong to Group I NPV. The HcGV genome was 114,557 bp with a 39% G+C content and contained 130 putative ORFs of 50 or more amino acids. When phylogenetic relationships were analyzed, HcGV was closely related to Xestia c-nigrum granulovirus, which belong to the Type-II GV. HcNPV shares 48 ORFs with HcGV. The most significant difference between HcNPV and HcGV is fgf gene. HcNPV contains one fgf gene, whereas HcGV contains three fgf genes. The presence of fgf reduces the time and efficient systemic infection it takes the virus to kill its host. The difference of fgf number from HcNPV and HcGV suggested that different affect for the speed of systemic infection.

Dielectrophoretic filtering and alignment of single-walled carbon nanotubes (SWCNTs) were tested using deuterium oxide as a solvent. A solution of deuterium oxide-SWCNTs was dropped on top of a silicon chip and an ac electric field was applied between pre-defined electrodes. Deuterium oxide was found to be a better solvent than hydrogen oxide for the dielectrophoresis process with higher efficiency of filtering. This was demonstrated by comparing Raman spectra measured on the initial solution with those measured on the filtered solution. We found that the aligned nanotubes along the electric field were not deposited on the substrate but suspended in solution, forming chain-like structures along the field lines. This so-called pearl chain formation of CNTs was verified by electrical measurements through the aligned tubes. The solution was frozen in liquid nitrogen prior to the electrical measurements to maintain the chain formation. The current-voltage characteristics for the sample demonstrate the existence of conduction channels in the solution, which are associated with the SWCNT chain structures.

To identify subspecies and stocks of minke whale meats purchased from Korean markets during 2005-2007, we first obtained their complete sequences of mitochondrial DNA cytochrome b and control region sequences, and compared these sequences to the corresponding sequences of the common minke whale (Balaenoptera acutorostrata), obtained from GenBank. From analyses with partial cytochrome b sequences (383 bp) and non-coding, partial control region sequences (463 bp), Korean mink whale meats are identified as products from the North Pacific minke whale (B. a. scammoni). In addition, the sequences of the partial control region from these meats showed G at site no. 298 and G or A at site no. 463, and the meats appeared to originate from the J stock within this subspecies. Thus, because the J stock has been protected since 1986, implementation of strict regulation measures to reduce their accidental fisheries by catch seems urgent. In addition, B. a. scammoni is distinct from B. a. acutorostrata, with an average Jukes-Cantor distance of 2.21% in the complete control region sequence analysis (935 bp) and 1.31% in the complete cytochrome b gene sequence analysis; the current results support the current subspecies classification, although further sequencing analyses with nuclear genes are necessary.

Cryopreservation of avian semen is a useful tool to preserve genetic resource for aim of preventing extinction induced by infectious disease like avian influenza. Unlike those of mammals, data from chicken cryopreserved semen has not been showed feasible results. So, various cryoprotectants and diluents have been examined in many methods. In this report, as a major ingredient of avian seminal plasm, glutamine was substituted by alanyl glutamine to enhance physiological stability of chicken semen during freezing. We studied effect of glycerol and Dimethylacetamide(DMA) on motility and progressive motility of spermatozoa using glutamine diluent(EK-G) or alanyl glutamine diluent(EK-A) condition. The semen of Ogye was collected twice a week by the dorso-abdominimal massage method and diluted with same volume of EK-G or EK-A at 25℃ and stored for 10 min at 4℃ in cold chamber. Glycerol or DMA was added to diluted semen to reached 7% of final concentration at 4℃. After 3min of equilibration, the diluted semen was packed into 0.25ml straws and subjected to cryopreservation used freezing equipment. The packed straw were placed on height 5 cm above surface of liquid nitrogen(LN2) and held for 10min. After preserved for 2 weeks, the straw was thawed onto the 4℃ cooling bath. The images of motility and progressive motility spermatozoa were recorded by digital image recorder and analyzed by manual. The results showed 68.5% motility and 34.1% progressive motility in DMA/EKA diluent, 31.45% and 17.6% in glycerol/EKA, 45.4% and 8.6% in DMA/EKG, and 9.7% and 6.4% in glycerol/EKG. With these results, the alanyl glutamine and DMA could be used as a main composition of diluent and cryoprotectant for cryopreservation of chicken semen.

The green peach aphid, Myzus persicae Sulzer, is one of the most important pests affecting protected and open-grown crops, because they cause direct damage by feeding on crops and indirect damage as virus vectors. It has recently become a serious problem because of the continuous use of insecticide resulting in resistance among green peach aphid population. Thus, the development of entomopathogenic fungi as aphid biocontrol agents has received increasing interest as part of integrated control strategies. In this study, we report the screening result of pathogenic fungi for the control of green peach aphid. Initial screenings were performed using 347 isolates of putative pathogenic fungi from Korea soils. As results, 20 isolates of entomopathogenic fungi were isolated from cadavers of green peach aphid supporting fungal conidiation. These isolates were identified as three strains of Lecanicillium attenuatum, nine strains of Beauveria bassiana, one strain of Metarhizium anisopliae, one strain of Metarhizium flavoviride, five strains of Paecilomyces lilacinus, one strain of Aspergillus sp. by microscopic examination, genetic sequencing of the ITS region and Universally Primed PCR (UP-PCR). Based on the screening results, twenty isolates were tested for their pathogenicity against adult green peach aphid. All fungal isolates were pathogenic to green peach aphid but mortality varied with isolates. These entomopathogenic fungi may be useful to develop eco-friendly insecticide to control green peach aphid.

Among 154 putative ORFs of Autographa californica multiple nucleopolyhedrovirus (AcMNPV), ac78 and ac79 are highly conserved genes in baculovirus, but their functions in the virus life cycle have been unknown so far. To determine their roles in AcMNPV replication, knockout mutants, ac78KO and ac79KO, were constructed using the plasmid capture system (PCS). Real-Time PCR analysis showed that both of ac78 and ac79 transcripts were first detected at 6 hours post-infection, and accumulated to maximum at 24 hours post-infection, suggesting that both of ac78 and ac79 are belong to late gene. When the genomic DNA of ac78KO was transfected into Sf9 cells, viral replication was restricted to a single cell infection. These results demonstrated that the ac78 play an important role in BV production, and therefore is essential for AcMNPV to mount a successful infection. Whereas Sf9 cells infected with the ac79KO showed normal viral symptoms such as rounding and swelling, OBs were not observed from majority of infected cells. These results suggested that the ac79 might play an important role in OB production.