본 연구에서는 용액 캐스팅법을 이용하여 각기 다른 함량의 VMT가 첨가된 SPAES/VMT 복합막이 제조되었다. SPAES 매트릭스 내의 VMT입자 분산은 전자주사현미경으로 관찰된 평균분포에 의해 확인되었다. 1 wt%보다 적은 함량을 포함한 복합막은 고분자 매트릭스 내에 좋은 분산성을 나타내어 막의 상부층과 하부층에 매끈한 표면을 가졌다. 복합막의 함수율은 온도가 증가함에 따라 급격하게 증가되었으며 VMT는 높은 이온교환능력으로 인하여 강한 수분친화도를 가짐으로 인해 높은 모든 흡착된 수분은 bound water인 것으로 확인하였다. VMT의 함량이 1 wt%보다 적게 첨가된 복합막에서 증가된 이온전도도와 낮아진 메탄올 투과도를 확인할 수 있었다. 모든 복합막 중에서 SPAES/VMT 1.0 wt% 복합막은 선택도 측면에서 가장 뛰어난 연료전지 성능을 가졌으며 Nafion 112과 비교하여 두배 이상 우수한 값을 나타내었다. 이것은 SPAES/VMT 1.0 wt% 복합막이 직접메탄올 연료전지의 구동을 위한 가장 우수한 조건이 될것이라 사료된다.

본 연구는 환경스트레스 저항성이 증진된 페튜니아를 개발하기 위하여 NDPK2유전자 도입 형질전환 계통 NDPK2-7-1와 SOD2 유전자 도입 형질전환 계통 SOD2- 2-1-1-35간의 교잡에 의해 획득된 후대들의 비생물적 스트레스 저항성을 조사하기 위해 수행되었다. 비 생물적 스트레스 유발원인 메틸바이올로젠(methyl viologen, MV) 100 μM과 200 μM 처리에서 교잡후대들은 그들의 교배 모본 SOD2 유전자나 NDPK2 유전자가 단독으로 도 입된 형질전환 계통이나 비형질전환체 보다 메틸바이 올로젠에 의한 피해를 적게 받았다. 이는 SOD2 유전 자나 NDPK2 유전자가 단독으로 도입된 형질전환 계 통간 교잡에 의해 획득된 후대들이 그들의 교배모본 (SOD2 유전자나 NDPK2 유전자가 단독으로 도입된 형질전환 계통)이나 비형질전환체 보다 산화적 스트레 스에 대한 저항성이 증진되었음을 증명해 준다고 할 수 있다. 이들 교잡후대들은 초장 등 11종류의 양적형질의 특성이 비형질전환체에 비해 약간 길거나 짧긴 하였지 만 비형질전환체와 거의 유사하였으며, 꽃 색갈이나 모양 또한 그들의 교배모본 (SOD2 유전자나 NDPK2 유전 자가 단독으로 도입된 형질전환 계통)이나 비형질전환 체와 차이가 없었다.

The purpose of this study is to see the effect of functional electrical stimulation on forced vital capacity and alternating motion rate in children with spastic cerebral palsy. This study divided 20 children with spastic cerebral palsy into two groups; functional electrical stimulation treatment group and control group. Functional electrical stimulation treatment group had 20min per day treatment three times a week for four weeks and the control group did not have any treatment. Before and after intervention, this study measured forced vital capacity and alternate motion rate(/peo/,/teo/) for all children. Forced vital capacity showed statistically significant increase for the group with functional electrical stimulation(p<.05) while the control group did not show any significant increase(p>.05). Alternate motion rate showed statistically significant increase for the group with functional electrical stimulation(p<.05) while the control group did not show any significant increase(p>.05). This result shows that functional electrical stimulation affected the ability of the children with spastic cerebral palsy who have decreased breathing and phonation capability.

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.

Peroxiredoxin V, an atypical thioredoxin peroxidase, is widely expressed in mammalian tissues. In addition, Prdx V is localized in mitochondria, peroxisome, cytosol, and nucleus. Prdx V has been reported to protect a wide range of cellular environments as antioxidant enzyme, and its dysfunctions may be implicated in several diseases, such as cancer, inflammation, and neurodegenerative disease. Identification and relative quantification of proteins affected by Prdx V may help identify novel signaling mechanisms that are important for oxidative stress response. However, the role of Prdx V in the modulation of hypoxia‐related cellular response is not studied yet. In order to examine the function of endogenous Prdx V in hypoxic condition in vivo, we generated a transgenic mouse model with Prdx V siRNA expression controlled by U6 promoter. Of many tissues, the knockdown of Prdx V expression was displayed in kidney, lung, and liver, but not spleen and skin. We conducted on the basis of nano‐UPLC‐MSE proteomic study to identify the Prdx V‐affected protein networks in hypoxic kidneys. In this study, we identified protein networks associated with oxidative stress, fatty acid metabolism, and mitochondrial dysfunction. Our results indicated that Prdx V affected to regulation of kidney homeostasis under hypoxia stress.

alcineurin (CN) is a calcium and calmodulin-depedent serine/threonine phosphatase. CN plays an important role in various biological processes including cell proliferation, cardiovascular, skeletal muscle development and apoptosis. In rheumatoid arthritis (RA), CN plays a role synoviocyte activation and arthritis progression. The selective inhibition of CN by the over-expression of CN-binding protein 1 (Cabin1). In the present study, joint restricted transgenic mice expressing the human Cabin1(hCabin1) were generated, driven by type II collagen promoter and efficiency of these mice was investigated by experimental arthritis. These transgenic mice successfully expressed hCabin1 in joint tissue as well as other organs like the liver, the heart, and the brain. The joint specific over-expression of hCabin1 reduced the disease severity during collagen-induced arthritis. In fibroblast-like synoviocytes (FLSs) from hCabin1 transgenic mice, the productions of these cytokines including, TNF-α, IL-1β and IL-6 were decreased and MMPs was also depressed in transgenic mice FLS. In addition, the expression of proapoptotic p53, p21, caspase-3, caspase-9 and Bax increased in transgenic mice, indicating that hCabin1 may induce FLS death by regulating the expression of Bcl-2, p53, p21, caspase-3, casepase-9 and Bax. It is expected that these findings will provide a more knowledge about the pathogenic mechanisms of rheumatoid arthritis and a potential animal model of the choronic inflammatory conditions, including atherosclerosis and transplantation.

Several types of white blood cells, such as T cells, B cells, and macrophages, are involved in the immune response. In particular, the processes of T-cell activation play a crucial role in an adaptive immune response, whereby the T-cell receptor (TCR) engages with an antigen and signals a cascade that leads to the activation of transcription factors (AP-1, NF-κB, and NFAT) that are critically involved in cytokine production. Roquin, encoded by the RC3H1 gene and characterized as an immune regulator, was recently identified as a novel RING-type ubiquitin ligase family member, but the mechanisms by which Roquin proteins regulate T-cell responses are unclear. To elucidate the role of Roquin in vitro, murine lymphoma EL-4 cells were used. Roquin overexpressing Tcells became hyper-responsive upon anti-CD3/CD28 stimulation in vitro and were a major source of cytokines such as IL-2, TNF-α, IL-6, and IL-10. Upon activation, these cells showed preferentially enhanced production of IL-2 and TNF-α, but not IFN-γ. To clarify the important role of Roquin in the T-cell response ex vivo, we generated T-cell-specific Roquin-transgenic (Tg) mice having a higher expression of Roquin in T cells as compared to wild-type mice. Using Roquin-Tg mice, we studied whether immune responses are affected ex vivo. Roquin-Tg CD4+ T cells showed enhanced production of IL-2 or TNF-α to TCR stimulation with anti-CD28 costimulation via up-regulation of CD28. T-cell proliferation also increased in Roquin-Tg CD4+ T cells after anti-CD3/CD28 treatment. Further studies on the role of Roquin in the regulation of primary T-cell activation, survival, and differentiation may be anticipated.

Testes‐derived unipotent male germ‐line stem (GS) cells can acquire multipotency under appropriate culture conditions to become mGS cells which can contribute to all three germ‐layers. This study was designed to investigate the epigenetic characteristics of mGS cells derived from adult mouse testes (maGS cells). The GS cells were isolated from 4 6 week DBA mouse and were cultured in Dulbecco’s modified Eagle Medium supplemented with 15% (v/v) fetal bovine serum, 1,000 U/ml LIF, 4 ng/ml GDNF at 37℃ in an humidified atmosphere of 5% CO2 in air to derive the maGS cells. The multipotency of maGS cells were verified by morphological and gene expression analyses, teratoma formation upon transplantation into nude mouse and in vitro differentiation ability. Bisulfite genomic sequencing revealed that GS cells had androgenetic DNA methylation pattern at the Igf2‐H19, Gnas‐Nespas , and Dlk1‐Dio3 imprinted gene clusters which changed to hemi‐zygotic embryonic stem (ES)‐cell like pattern in the maGS cells. Western blot analysis, using modification‐ and residue‐specific antibodies, revealed that both maGS and ES cells had similar level of histone di‐methylation at 4th and 27th lysine residue of histone 3 (H3K4me2 and H3K27me2) which represent “bivalent domain” for regulating self‐renewal and differentiation of mouse ES cells. Both maGS and ES cells also shared similar hisone modification for H3K9me2, H3K79me2, H3K9ac and H3K18ac. However, maGS cells had higher level of H3K- 36me2 and H3S10p. These data suggest that maGS and ES cells share several epigenetic characteristics but they also have their own unique epigenetic marks that may be useful as a molecular marker for their identification.

Nicotine, a major teratogen of cigarettes smoke induces embryonic abnormalities during the early stages of organogenesis. In this study, the protective effect of β-carotene against nicotine–induced embryos was evaluated by morphologic scoring, nile blue staining, lipid peroxidation, SOD activity assay and real-time PCR. The embryos exposed to nicotine (1 μM) revealed remarkable morphological anomalies compared to normal control group (p<0.05), but when β-carotene (1×10‒4 μM or 5×10‒4 μM) was added concurrently to the embryos exposed to nicotine, morphological parameters were significantly improved (p<0.05). Nicotine induced oxidative stress by increased lipid peroxidation, expression of proinflammatory cytokines (TNF-α and IL-1β), caspases-3 and decreased SOD activity. However, administration of β-carotene (1×10‒4 μM or 5×10‒4 μM) restored the SOD level and decreased oxidative damage in the embryos. These results indicate that β-carotene effectively counteracts the deleterious effects of nicotine on embryos and attenuates oxidative damage possibly through its antioxidant effects.

Nicotine, a major toxic component in tobacco smoke, leads to severe embryonic damages on organogenesis. We investigated if resveratrol can inhibit the nicotine–induced teratogenesis in the cultured mouse embryos (embryonic day 8.5) for 48 hours using a whole embryo culture system. The embryos exposed to nicotine (1 μM) revealed severe morphological anomalies, the increased levels of caspase-3 mRNA and lipid peroxidation, and further the lowered levels of mitochondrial manganese superoxide dismutase (SOD), cytosolic glutathione peroxidase (GPx), phospholipid hydroperoxide GPx, hypoxia-inducible factor 1α, and sirtuin mRNAs and SOD activity significantly compared to normal control group (p<0.05). However, whenre sveratrol(1×10‒5 μMor1 ×10‒4 μM) was added concurrently to the embryos exposed to nicotine, these all parameters were significantly improved (p<0.05).These findings indicate that resveratrol has a protective effect against nicotine-induced teratogenesis in mouse embryos throughout antioxidative and anti-apoptotic activities.

The objective of this study was to evaluate in vitro production of bovine embryos in Hanwoo. Oocytes were collected by ovum pick up (OPU) from ovaries of genetically high-value Hanwoo or by needle puncture from ovaries of slaughtered cattle. OPU was done every 3 4 days duing experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, We compared the in vitro maturation rate in two groups (Experiment 1). 545 oocytes were recoverd from 4 females by 32 trials of OPU and then 433 oocytes were shown MⅡ stage after in vitro maturation (79.4%). In case of needle puncture group, 1905 oocytes were collected and then 1420 oocytes were matured to MⅡ stage during in vitro culture(74.5%). Second, we compared the developmental rate to blastocyst in two groups (Experiment 2). 1420 oocyte by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24 48 h after in vitro fertilization (IVF) was 88.6% and blastocyst development rate was 20.5% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (84.8%), blastocyst development rate was higher compared with needle puncture group (26.4%). In conclusion, Blastocyst developmental rate could be increased by OPU than classical method of needle puncture. Improvement of bio-technique in collecting oocytes could be applied to understand the reproductive physiology in cattle, expecially Hanwoo. Therefore, further investigation should be done to clarify the efficiency and advantage of OPU involved in reproduction in animals and human being. This research was suppoted by Imsil-gun agricultural technology service center.

The cotton aphid, Aphis gossypii (Glover), is one of the most serious pest in the cultivation of various vegetables. A highly imidacloprid-resistant field population (CA-L) was collected from cucumber at Gangwha island in 4th August 2011. Even though neonicotinoid insecticides especially imidacloprid were sprayed six times during June and July, aphid density was too high to be counted. IEF and 2DE analyses revealed that general esterase isozyme (pI. 5) in CA-L were dramatically overexpressed and more isozyme spots identified in CA-L compared to susceptible (CA-S) strain. To identify differentially expressed genes in CA-L, comparative transcriptome analyses based on GS-FLX were conducted with total RNA extracted from CA-L, which generated ca. 143 Mb reads. Previously reported, comparative transcriptome analyses performed in imidacloprid resistant (CA-IR) and CA-S. The comparative transcriptome analyses re-investigated after all data sets were combined together. As previously reported, seven ATP-binding cassette (ABC) transporter genes were newly identified in A. gossypii, among which only ABCC9 gene was highly expressed in CA-IR and L. These results suggested that ABCC subfamily associated with imidacloprid resistance in A. gossypii.

Historically the rise of contagious diseases and epidemics has been mainly caused by lack of food safety knowledge and negligence in personal hygiene. Food poisoning is an example that have been caused by lack of proper knowledge in food safety and personal hygiene. In order to prove this hypothesis, our team distributed a total of 225 survey sheets to middle school students. Survey questions mainly focused on the students' food safety knowledge, personal hygiene and regular dietary habits. We used 12 items to determine the stages of change in teenagers and divided into 3 stages of change: precontemplation stage (poor problem recognition), contemplation and preparation stage (some problem recognition), and action stage. Out of the 225 students surveyed, 105(47.1%) were male and 118 (52.9%) were female; 44.0% were in 7th grade, 33.0% in 8th grade, and 24.0% in 9th grade. In stages of change model, 3.8% were classified in pre-contemplation stage, 31.1% in preparation stage, and 65.1% in action stage. The awareness of the significance of food safety has a strong positive correlation with the stages of change food safety behavior (P<0.001), which suggesting our questionnaires regarding stages of change and food safety behavior were quite credential. Stages of change were also correlated with the consumption of unsanitary foods (P<0.01) and with the frequent hand washing (P<0.05). The stages of change showed a positive correlation with the food safety knowledge level (P<0.05). The four questions (out of 10) with the least percentage of correct answers were questions about the meaning of Norovirus (23.6%), the meaning of expiration date (43.6%), the meaning of food poisoning (36.4%), and proper food-handling (36.0%).

Zinc oxide nanoparticles (nZnO) are used in a various range, including ceramic manufacture, photocatalysis, UV filters, and the food industry. However, little is known about the effects of micro- and nano-particles during mouse embryo organogenesis. To determine whether ZnO affects size-dependent anomalies during embryonic organogenesis, mouse embryos were cultured for two days with 300 ug/ml micro ZnO (mZnO;80±25 μm) and nZnO (< 100 nm) and the developmental changes were then investigated. Quantity of Zn by inductively coupled plasma mass spectrometry analysis, and expression patterns of various antioxidant enzymes in the embryos were investigated. Embryos exposed to mZnO or nZnO exhibited severe retardation of growth and development. In embryos exposed to mZnO and nZnO, yolk sac diameter, crown-rump length, and head length were significantly diminished. The morphological parameters, including yolk sac circulation, allantois, flexion, heart, hindbrain, midbrain, forebrain, otic system, optic system, branchial bars, maxillary process, mandibular process, olfactory system, caudal neural tube, forelimb, hindlimb, and somites in mZnO and nZnO-treated groups were significantly decreased. Zn absorption of the nZnO-treated group was significantly higher than that of the mZnO-treated group. Significantly decreased levels of CuZn-SOD, Mn-SOD, cGPx, and PHGPx mRNA were observed in the ZnO-treated group. In addition, antioxidant enzyme mRNA expressions of the nZnO group were significantly diminished, less than those of the mZnO treated group. These findings indicate that 300 ug/ml ZnO showed abnormality and nZnO may have a more severe effect than mZnO in developing embryos.

Porcine Circovirus Type2 (PCV2), a single-stranded DNA virus associated with Postweaning multisystemic wasting syndrome(PMWS) of swine, has two major open reading frames, ORF1 and ORF2. The genomic size and molecular weight of ORF2 is respectively 699bp, 28kDa. ORF2 encodes the capsid protein (structural protein) that has type-specific epitopes and is very immunogenic and associated with the induction of neutralizing antibodies, suggesting its potential use in diagnostic assays as well as vaccine development. For efficient production of the capsid proteins, we expressed the PCV2 ORF2 gene with baculovirus in the insect cells. In this study, PCV2 ORF2 was appropriately ligated into the baculovirus transfer vector, pBacPAK9 and pB9-Acpol19-110-EK. Sf21 cells were transfected with a mixture of the purified recombinant transfer vector and bAcGOZA. We generated and purified recombinant viruses containing PCV2 ORF2, and named rAc-B9-PCV2ORF2 and rAc-B9-19-110-EK-PCV2ORF2, respectively. Expression levels of capsid fusion proteins with a partial polyhedrin region of AcNPV more increased than recombinant proteins from non-fusion expressed. Also, expression efficiency increased over time and differed at MOI. As a results, fusion expression of porcine circovirus type2 ORF2 using baculovirus could be utilized as an alternative expression method to produce recombinant antigen against PCV2 infection and is worthy of further investigation.