A rapid cold hardening (RCH) and supercooling capacity usually play crucial roles in survival during the overwintering period in the tobacco budworm, Helicoverpa assulta, a freeze-susceptible species. Cryoprotectants such as polyol or sugar affect RCH and maintain fluid status of hemolymph. This study is performed to identify cryoprotectants as a RCH factor in H. assulta. Pre-exposure of H. assulta larvae to 4℃ significantly increased survival at -10℃ in all developmental stages from egg to adult. RCH was dependent on the duration of the pre-exposure period. RCH also significantly enhanced the supercooling capacity. Analysis of cryoprotectants from the hemolymph revealed that the pre-exposure treatment allowed the larvae to accumulate glycerol and trehalose among polyols examined. In addition, unknown materials from 2 peaks were also increased. TIC analysis revealed 3 predicted formulas for unknown materials, C26H24O20S or C3H4N6OS and C22H20O21. Glycerol-3-phosphate dehydrogenase (GPDH) and glycerol 3-phosphatase (G3P) that involving in glycerol biosynthesis were identified from the transcriptome of H. assulta 4th instar larvae. Based on the expression level of transcripts, the expressions of GPDH and G3P were relatively increased when compared to that of the control, suggesting that these genes contribute to overwintering and biosynthesis of glycerol.

The present research investigated copper and cadmium stress-induced differentially expressed genes (DEGs) using annealing control primers (ACP) with the differential display reverse transcription polymerase chain reaction technique in alfalfa (Medicago sativa L. cv. Vernal) leaves. Alfalfa leaves were subjected to 250 μM of copper and cadmium treatment for a period of 6 h. A total of 120 ACPs was used. During copper and cadmium treatment, 6 DEGs were found to be up or down regulated. During copper stress treatment, 1 DEG was up-regulated, and 3 novel genes were discovered. Similarly, during cadmium stress treatment, 1 DEG was up-regulated and 5 novel genes were identified. Among all 6 DEGs, DEG-4 was identified as the gene for trans-2,3-enoyl-CoA reductase, DEG-5 was identified as the gene for senescence-associated protein DIN1 and DEG-6 was identified for caffeic acid O-methyltransferase. All the up-regulated genes may play a role in copper and cadmium stress tolerance in alfalfa.

This study was conducted to evaluate the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To determine the movement of degradable HGC nanoparticles in the body, 20 mg/kg of lutetium177-labeled HGC (Lu177-HGC) with the size ranging from 320 to 400 nm was injected intravenously into ICR mice, and the amount of radioactivity remaining in blood and several organs was measured at various time points during the period of 5 days. In the pharmacokinetics analysis using the Lu177 radioisotope, the free Lu177 was mainly distributed and accumulated in the order of kidney>liver>lung at 1 day after the injection of the radioisotope. However, the Lu177-HGC showed a high distribution of nanoparticles in the order of liver>spleen>kidney during the experimental period of 5 days. These results would provide a basic pharmacokinetics for the use of HGC as a drug carrier in drug delivery system.

The ultimate goal of this study is to assess the accumulation and distribution of hydrophobically modified glycol chitosan (HGC) as a degradable nanoparticle in the body. To understand the movement of degradable nanoparticle HGC in the body, we intravenously injected a dose of 20 mg/kg of Cy5.5-labeled HGC with size ranging from 320 to 400 nm into ICR mice, and measured the amount of fluorescence remaining in blood and several organs at various time intervals. In blood, the level of Cy5.5-labeled HGC was the highest at 15 min, then after 30 min it decreased rapidly and reached a plateau form 30 min to 28 days. In the tissue we confirmed the presence of nanoparticles at high levels in the order of kidney>liver>submandibular gland until 28 days after injection. However, we did not find the presence of the particles in the brain or testes. These results will provide basic information on HGC as a drug delivery agent.

The pheromone biosynthesis in Plutella xylostella is more active in the scotophase than in the photophase, which suggests that there may be changes of gene expression in the pheromone glands. To identify genes contributing to change in pheromone production, we analyzed transcriptomes from pheromone glands of both decapitated females in the photophase and normal ones in the scotophase. Comparative analysis were performed with transcriptomes of pheromone glands from non-decapitated (PG) females and decapitated ones for identification and expression of putative genes associated with pheromone biosynthesis pathway. Deep sequencing for mRNAs in the pheromone gland yielded approximately 7.5Gb and totally 17265 transcript were constructed under a homology cutoff of 10-6 Evalue. Genes putatively involved in pheromone biosynthesis were identified such as acetyl-CoA carboxylase, acetyl-CoAdehydrogenase, fatty acid synthase (FAS), desaturases (Δ9 and Δ 11) and fatty acid reductases (FAR) including pgFAR, alcohol oxidase, aldehyde oxidase and aldehyde reductase, etc. Expression of 6 signal genes involving in pheromone biosynthesis such as acyl-CoA desaturase, FAR, PBAN receptor, fatty acid transporter, acyl-CoA binding protein did not exhibited ant significant different in both transcriptomes. Quantitative RT-PCR revealed that expressions of FAS, Δ11 desaturase and pgFAR were higher in PG than that in ΔPG. Based on results, Δ11 desaturase and pgFAR may have a crucial role in sex pheromone biosynthesis of P. xylostella.

Based on dual coding theory, this study predicted that pictorial typography (PT), a device involving direct visual integration of a written word into a picture of its referent, is more effective for ESL children’s pre-literacy skills, than the non-integrated paring of word and picture on flashcards (FC). The two types of instruction were given to thirty-two Korean children aged six and seven, both embedded within the WordWorld (WW) episodes, an animated TV series developed by PBS. Results from the pre- and post-test design were as follows. PT was more beneficial than FC for the two upperlevel skills, vocabulary and reading fluency, while they were equally beneficial for the two lower-level skills, letter recognition and phonemic awareness. The two mediating variables, gender and age, were shown to have partial effects as well. Dual coding theory and the importance of instructional intervention have been supported by this study.

The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15. The cultivation methods of Acanthopanax in Korea need to be optimized. Hence, this study investigated the effect of different fertilizer ratios and planting dates on the growth and acanthoside D content of two (2) Acanthopanax species. The current recommended fertilizer rate of 10.5-8.5-8.5 kg/ha- (N-P2O5-K2O, respectively) produced the best plant growth of Acanthopanax. For the first year, the acanthoside D content resulting from the 2P (2x phosphate) rate was higher than that from the other fertilizer ratios, yet there were no significant differences resulting from the various treatments for either Acanthopanax divaricatus or Acanthopanax koreanum. Similarly, for the second year, there were no significant differences in the acanthoside D content resulting from the various fertilizer ratios, although for both species the acanthoside D content resulting from the 2P rate was slightly higher than that from the other treatments. Therefore, the results indicated that doubling the amount of phosphate increased the acanthoside D content. Plus, the optimum planting date with respect to growth and productivity for Acanthopanax divaricatus was identified as April 15.

Matrix metalloproteinases (MMPs) have been known to affect to cell migration, proliferation, morphogenesis and apoptosis by degrading the extracellular matrix. In the previous studies, undifferentiated mouse embryonic stem cells (ESCs) were successfully proliferated inside the extracellular matrix (ECM) analog-conjugated three-dimensional (3D) poly ethylene glycol (PEG)-based hydrogel. However, there is no report about MMP secretion in ESCs, which makes it difficult to understand and explain how ESCs enlarge space and proliferate inside 3D PEG-based hydrogel constructed by crosslinkers containing MMP-specific cleavage peptide sequence. Therefore, we investigated what types of MMPs are released from undifferentiated ESCs and how extracellular signals derived from various niche conditions affect MMP expression of ESCs at the transcriptional level. Results showed that undifferentiated ESCs expressed specifically MMP2 and MMP3 mRNAs. Transcriptional up-regulation of MMP2 was caused by the 3D scaffold, and activation of integrin inside the 3D scaffold upregulated MMP2 mRNAs synergistically. Moreover, mouse embryonic fibroblasts (MEFs) on 2D matrix and 3D scaffold induced upregulation of MMP3 mRNAs, and activation of integrins through conjugation of extracellular matrix (ECM) analogs with 3D scaffold upregulated MMP3 mRNAs synergistically. These results suggest that successful proliferation of ESCs inside the 3D PEG-based hydrogel may be caused by increase of MMP2 and MMP3 expression resulting from 3D scaffold itself as well as activation of integrins inside the 3D PEG-based scaffold.

본 연구에서는 심지층 처분장 부지선정 시에 고려되는 요소를 지질, 수리지질, 지화학 등으로 분류하고 그 첫 번째 단계로 지질분야의 세부 항목을 지형, 토양층, 암종, 구조지질, 역학적 안정성, 지질학적사건으로 분류하였으며, 이들 항목에 대한 국외 기준분석을 수행하였다. 부지선정요소(Siting factor)에 대한 기준(Criteria)은 각 국가의 처한 지질환경에 따라 다른 조건혹은 값을 제시하고 있다. 화산 및 지진활동이 빈번한 일본에서는 이에 대한 기준을 상대적으로 자세히 기술하고 있으며, 빙하작용이 예상되는 스웨덴에서는 빙하작용에 의한 지반 융기·침식에 대한 영향을 상세히 분석하였다. 따라서, 본 논문 결과는 향후 국내의 심지층 처분장 부지선정 기준 수립시에 중요한 참고자료로 활용될 수 있을 것으로 판단된다.

Semen can be divided into two parts. One is cellular part which contains sperms the other is liquid part which is called by seminal plasma. The seminal plasma is a nutritive and protective medium for the sperms. Fructose, which is major energy source, is supplied to sperms swim to female oocyte. Alkalic property protects sperms from hostile environment of female reproductive organ. Also, seminal plasma induces tolerance to preexisted immune cells, and changes intra‐uterine environment to better conditions for fertilized embryos to implant. However, the effects of seminal plasma in in vitro culture of fertilized embryos are unclear. Second fraction of fresh semen was obtained from a normal farm pig. The semen was centrifuged to remove sperms, and then supernatant was filtrated. The filtered seminal plasma was stored in — 30℃. In this study, electrically activated and chemically activated porcine embryos were employed to investigate the developmental rate after 2 hours treatment of none, 0.1%, 0.5%, and 1% seminal plasma in culture media by two days of activation. Both electrically and chemically activated embryos, cleavage rate and cell numbers of blastocysts were not significant difference within four groups. Blastocyst formation rate of electrically activated embryos also did not show significant difference within any groups. However 0.1% seminal plasma treatment group showed significantly increase of blastocyst formation rate in chemically activated group (None; 24.8%, 0.1%; 31.7%, 0.5%; 19.4, and 1%; 16.5%, respectively. p<0.05).

The purpose of this study is to examine the effects of gait training using functional electrical stimulation on the improvement of hemiplegic patients' functions for balance and gait velocity. The subjects of the experiment were determined to be 10 each hemiplegic patients who had been diagnosed with stroke or brain damage six months or longer earlier assigned to an experimental group and a control group respectively. The subjects were evaluated before the experiment using Tetrax and 10M gait tests, received gait training five times a week for four weeks using functional electrical stimulation and were evaluated after the experiment in the same method as used in the evaluation before the experiment. In order to examine differences between the experimental group that received gait training using functional electrical stimulation and the control group that was treated by functional electrical stimulation and received gait training thereafter, differences between before and after the experiment were analyzed using paired sample t-tests and differences in changes after the experiment between the experimental group and the control group were analyzed using independent sample t-tests in order to compare the two groups with each other. Experimental results showed significant differences in weight bearing, balance and gait velocity between before and after the experiment in the experimental group(p<.05). In the control group, whereas weight bearing and gait velocity did not show any significant difference between before and after the experiment(p>.05), balance showed significant differences(p<.05). Weight bearing, balance and gait velocity change rates showed significant differences between the experimental group and the control group(p<.05). In conclusion, it was indicated that gait training using functional electrical stimulation is effective for enhancing stroke patients' weight bearing rates, balance abilities and gait velocity.

The green peach aphid (Myzus persicae) is a cosmopolitan pest of agricultural and horticultural crops and causes serious economic damages. M. persica has rapidly developed resistance to a wide variety of insecticides, including pyrethroids. Target site insensitivity mechanism mediated by two mutations (L1014F and M918T) on the para-type voltage-sensitive sodium channel (vssc) is mainly responsible for pyrethroid resistance. To predict the vssc resistance allele frequency, quantitative sequencing (QS) protocol was established. Frequency prediction equations generated from the plots of signal ratios and amplification critical time showed a high correlation coefficient (r2>0.993), indicating its high accuracy in prediction. QS results revealed that the kdr-type L1014F mutation is only present in Pyeongchang strain. No field strains of M. persicae possessed the super-kdr type M918T mutation. However, a novel M918L mutation was found by genotyping approach. The allele frequencies of M918L and L1014F were 0% to 53% in populations examined, and the level of M918L mutation frequency was closely related with pyrethroid resistance. Therefore, QS-based detection of M918L mutation frequency should faciltate the monitoring of pyrethroid resistance in the field.

The green peach aphid (Myzus persicae) is a serious pest of agricultural and horticultural crops all over the world. M. persica has rapidly developed resistance to a wide variety of insecticides, including carbamates. The E4/FE4 carboxylesterase is known to be involved in carbamate resistance. To compare the E4/FE4 carboxylesterase gene copy number, as a genetic resistance marker, between seven field strains, quantitative real-time PCR (qPCR) was performed. In addition, quantitative sequencing (QS) was employed to predict the frequencies of acetylcholinesterase (AChE) mutations (A301S and S431F) that are associated with target site insensitivity. All M. persica strains examined possessed the S431F mutation in the heterozygous state except for a susceptible strain, implying the possibility of AChE duplication. In contrast, no A301S mutation was found. Frequency prediction equation was generated from the plots of signal ratios and amplification critical time, which showed a high correlation (r2>0.996). QS analysis of M. persicae populations revealed that the allele frequency of S431F ranged 4% to 63%. Taken together, the AChE resistance allele frequencies determined by QS and the E4/FE4 gene copy number by qPCR should facilitate the detection and monitoring of carbamate resistance in M. persicae in the field.

Rhynchium brunneum is a widely distributed wasp species in South Eastern Asia. R. brunneum females were collected from rural provinces of Cambodia, and their total RNA and venom were extracted on site. To search for novel substances in venom, a subtracted cDNA library specific to the venom gland and sac was constructed. A total of 1118 expressed sequenced sequence tags (ESTs) were sequenced and assembled into 349 contigs (107 multiple sequences and 242 singletons). In this result, we found the putative neurotoxin (DTX protein precursor), antimicrobial peptides (teratocyte-specific caboxylesterase) together with typical major components of wasp venom (venom hyaluronidase, arginine kinase, phospholipase A2, serine/theonine protein phosphatase). Additional in-depth annotation would be required for further characterization of many unidentified genes found in the EST library.

Vespa tropica is a tropical species of Vespa found in Southeast Asia. V. tropica wasps were collected from rural provinces of Cambodia, and their total RNA and venom were extracted on site. To search for novel substances in venom, a subtracted cDNA library specific to the venom gland and sac was constructed and venom protein was analyzed by nano-LC-MS/MS. A total of 1127 expressed sequenced sequence tags (ESTs) were sequenced and assembled into 572 contigs (152 multiple sequences and 420 singletons). The short venom peptides were identified to be encoded from 5 contigs (43 ESTs) by proteomic analysis. In addition, putative antimicrobial peptides together with typical major components of wasp venom (venom allergen 5, mastoparan-like peptide, serine protease, and hyaluronidase) were identified in the EST Library. Additional in-depth annotation would be required for further characterization of many unidentified genes found in the EST library.

This study aims to reveal how EA affects BAX and NF-kB involved in cell deaths from global ischemia, and to do this, observes the changes of BAX and NF-kB caused by EA application after transient global ischemia. The experimental method is to give rise to global ischemia and apply EA to 27 SD rats with the particulars of being six-week-old, male, around-300 gram-weighing, and adapted to laboratory environment for more than a week, and divide them into three groups, that is, GV20 EA group(n=9), L14 EA group(n=9), no-treatment GI group(n=9), and then observe their changes of BAX and NF-kB at the time lapse of 6 hours, 9 hours and 12 hours after ischemia, using western blotting. The numerical decrease of BAX expression at the time lapse of 9 hours after EA application, though not statistically significant, was observed in GV20 EA group and L14 EA group, and the NF-kB expression appeared statistically significant decrease in GV20 EA group and L14 EA group, but the expression was higher in the group with EA application. Therefore, EA application at the early phase of global ischemia is considered to affect BAX and NF-kB and play a positive role in decreasing apoptosis and cell deaths by inflammation.

Insects or insect remains found in beer are one of major issues in consumer claim. Accurate estimation of inflow time isa critical factor for the settlement of such claims related with beer-contaminating insects but no reliable methods have been developed. In an attempt to establish a molecular marker-based diagnostic method, the degradation rates of 18S rRNA genes in the insectssoaked in 500 ml beer were investigated by quantitative real-time PCR (qPCR) over one month period at room temperature. Among the six insect species tested, the house fly (Musca domestica) and honey bee (Apis mellifera) revealed high correlations (r2=0.974-0.990) between the degradation of 18S rRNA gene and inflow time. In these insects, statistically significant distinction was possible between the samples stored in beer less than 14 days and more than 14 days. Other insects, including the fruit fly, common house mosquito, German cockroach and Indian meal moth, displayed poor correlations, which appeared attributed to the inefficient genomic DNA extraction likely due to small sample size or disintegration of body parts during storage in beer. With proper improvement in DNA extraction, this 18S rRNA-based diagnostic method would be applicable for estimating the inflow time of beer-contaminating insects.