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).

Cathepsin B, a lysosomal cystein protease that plays an important role in the degradation of intracellular proteins in lysosomes, is detected in a wide variety of cells including bovine oocytes and embryos. Although the mode of action of cathepsin B is not fully understood, a strong relationship was observed between cathepsin B and apoptosis in many types of cells. Cathepsin B was found to induce the apoptotic pathway through activating initiator caspases rather than executioner caspases. Thus, the aim of this study was evaluated the effect of capthesin B inhibitor, E-64, on blastocyst developmental competence and subsequent preimplantation quality of the IVF and SCNT bovine embryos. After IVF and SCNT procedures, presumptive bovine embryos were cultured in CR1aa medium supplemented with E-64 for 24 h. Then, samples were additionally cultured in CR1aa medium without E-64 for 5 days. In our results, the frequency of blastocyst formation was higher when treated with E-64 compared with the control group (p<0.05). Furthermore, the blastocyst cell number was enhanced and apoptosis reduced (TUNELpositive nuclei number) by E-64 treatment in both IVF and SCNT bovine embryos (p<0.05). In the real-time quantitative RT-PCR, the expression of anti-apoptotic Bcl-xL gene was shown to be increased in the blastocyst stage, whereas expression of proapoptotic Bax was decreased. In conclusion, our results indicate that E-64 improves the developmental competence and embryonic qualities of bovine IVF and SCNT embryos by modulating cathepsin B induced apoptosis during the preimplantation stage.

Phosphoprotein Enriched in Astrocytes (PEA15) is a 15kD-sized intracellular signaling protein, highly expressed in astrocytes and constitutively expressed in peripheral tissues. Recently it was found that PEA15 expression was elevated in patients suffering type 2 diabetes and suggested to be involved in the syndrome of insulin resistance. To investigate the functional role of PEA15 for the control of blood glucose level, we produced a transgenic pig over-expressing mouse PEA15 (mPEA15). As a model animal, pig has many advantages. They have a higher fecundity and a short generation time and are physiologically similar to human. Using the transgenic pig, we carried out a series of experiments to establish a link between PEA15 expression and the insulin resistance. Our results suggested that, compared with control pig, mPEA15 pig has, (1) a higher blood resistin level, (2) a lower cell membrane-embeded GLUT4 level, and (3) a lower glucose clearing ability based on oral glucose tolerance test (OGTT). When our results combined, it can be concluded that mPEA15 over-expressing pig has many symptoms of insulin resistance and these pigs will become a useful disease model to investigate diabetes mellitus in the near future.

The purpose of this thesis is to examine the effect of hormone treatment in blastocyst development of in vitro cultured porcine oocyte. Oocytes used in the study was matured in vitro in the presence of 10% FBS or 10% pFF, and treated with FSH, LH or FSH+ LH, and the rate of blastocyst development was assessed based on the expression of autophagic genes. There was no significant differences in blastocyst development between oocytes maturaed in 10% FBS or 10% pFF. In vitro matured oocytes treated with FSH+LH showed blastocyst development rate as high as that of untreated oocytes, while groups treated with LH only showed a decrease in blastocyst development. About the expression of cell death assosiated factors, mRNA levels of autophagy and apoptosis genes were increased in oocytes matured in 10% FBS and treated with LH. Oocytes that did not receive hormone treatment showed low expression of most cell death genes except ATG5. When oocytes were matured in 10% pFF, ATG5 expression was the highest in FSH treated group, while LC3 showed strong expression in all hormone treated groups. On the other hand, the expression level of mTOR and caspase-3 did not show significant differences between groups. We also examined the protein level of apoptotic genes in the blastocyst. The amount of caspase-3 protein was similar between groups matured in 10% FBS and 10% pFF, but was the highest when treated with LH. Blastocysts treated with FSH and FSH+LH showed similar level of caspase-3 protein, while the level was the lowest when hormone treatment was not given. Within the blastocyst, caspase-3 was mostly expressed in trophoblast cells when matured in 10% FBS, while maturation in 10% pFF caused expression of this protein in the inner cell mass (ICM). Expression of MAP1LC3A was higher in groups matured in 10% pFF than groups matured in 10% FBS in all types of hormone treatment. Among the blastocysts matured in 10% pFF, MAP1LC3A level increased in the order of untreated < FSH < FSH+ LH. Expression of MAP1LC3A within the FBS-matured blastocyst was concentrated to the trophoblast, while pFF-matured blastocyst showed expression in both trophoblast and ICM. The expression of mTOR showed a pattern opposite to that of MAP1LC3A. That is, its expression was the lowest in 10% FBS group without hormone treatment. In both FBS and pFF group, and in all three combinations of hormone treatment, mTOR expression was ovserved mostly in ICM. Together, these results indicated that hormone treatments tend to induce expression of genes associated with programmed cell death. We suggest that proper induction of programmed cell death by FSH and LH treatment would increase the rate of blastocyst development. * This work was supported by BioGreen 21 Program (No. PJ008029). Rural Development Administation, Republic of Korea.

The objective of this study was to investigate the proteome composition in pretermand term‐derived human umbilical cord. Umbilical cord samples were collected from 6 preterm infants with gestational age less than 36 weeks or 4 full terms together with medical information during prenatal period. Several biomarkers are routinely used clinically for predicting preterm labor; however, these factors are either nonspecific or detected too late. Protein profiles were performed on samples from both preterm‐ and termderived human umbilical cord by using Two‐dimensional gel electrophoresis (2‐DE). Approximately 200 different proteins were identified between preterm‐ and term‐delivered umbilical cords. Among them, differentially expressed 34 proteins were identified in 48 protein spots. In the preterm‐derived human umbilical cords, 15 proteins were present at higher levels (2.0‐ to 9.28‐fold increases) and 19 were present at lower levels (2.0‐ to 11.8‐ fold decreases or not detectable) compared to the control term‐derived umbilical cords. Proteomics approaches such as 2‐DE could greatly facilitate the discovery of new and better biomarkers. The high sensitivity and specificity achieved by the combined use of the selected biomarkers show great potential for the early detection of Adverse pregnancy outcomes such as pre‐eclampsia, small for gestational age infants, preterm delivery and placental abruption are associated with higher mortality. Increased amount of HIF‐1 α, GAPDH and HSP27 were observed in preterm‐derived umbilical cords was due to hypoxia‐ dependant and oxidative stress‐independent manner. Moreover, we isolated HUVEC (Human umbilical vascular endothelial cells) from preterm‐ and term‐derived umbilical cords and examined LDH activity. The results of the current study may provide important insights into the molecular mechanisms underlying umbilical cord development and also these data will contribute to a better understanding of the composition of preterm‐ and term ‐ derived human umbilical cord and aid the discovery of novel biomarkers for the prenatal diagnosis of fetal abnormalities

Light-emitting diodes are used in applications as diverse as aviation lighting, automotive lighting, advertising, general lighting, and traffic signals. LED panel light source used in an experiment on plant and mushroom growth. To establish the optimum artificial light illumination method for Pleurotus eryngii in closed mushroom factory system, the effects of red LED light quality on growth was investigated. The humidity was maintained more than 90% and temperature was 15~16℃, and CO2 density was 500ppm. The fresh weightand height were higher under red LED light (650nm, 73.1 Pmol/m 2s) comparing with control (fluorescent lamp, 10~20 lux). β-aminoisobutyric acid and total phenol content were higher under red LED light (650nm, 73.1 Pmol/m2s).

The resistance levels to carbamate (CB) and organophosphate (OP) insecticides were determined by topical application in 14 field strains of Nilaparvata lugens. The resistance levels to CB and OP were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. A quantitative sequencing (QS) protocol was established to determine the allele frequencies of four acetylcholinesterase point mutations putatively associated with CB and OP resistance. The allele frequencies of the four mutations (G119A, F/Y330S, F331H and I332L) in field strains ranged from ca. 0.0~51.7%, 0.0~88.9%, 2.5~47.7%, 5.1~56.0% and 6.7~57.3%, respectively. The F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with actual resistance levels (r2= < 0.232), whereas F331H and I332L showed a better correlation with the resistance levels of benzofuranyl methylcarbamates (r2= 0.595). This finding indicates that F331H and I332L mutation frequencies may be used as molecular markers for detecting carbamate resistance in N. lugens. A QS protocol detecting the F331H and I332L mutation frequencies would be employed as a supportive tool for rapid monitoring of CB insecticide resistance levels in N. lugens

The screening of effective lethal genes was conducted via the systemic delivery of dsRNA for the RNA interference (RNAi)-based management of Tetranychus urticae. Four candidate genes (coatomer protein complex, T-COP; M1 metalloprotease, T-M1MP; Ribosomal protein S4, T-RPS4; A subunit of V-ATPase, T-VATPase) with a reference genes (EGFP) were tested for the experiment. Translocated dsRNA to the leaf disc (ca. 30 mm diameter) was detected at 12 h post-treatment, indicating that dsRNA could move through leaf veins. In the evaluation of mortality, the T-COP, T-RPS4 and T-VATPase exerted moderate levels of toxicity at 72 h post-treatment among four dsRNA tested. Knockdown of the target genes were confirmed by quantitative PCR, suggesting that RNAi of target genes was accomplished by feeding dsRNA. In summary, the dsRNA delivery via leaf disc was effective in screening lethal genes, and some genes, such as T-COP, T-M1MP, T-RPS4 and T-VATPase, can be applicable for establishing a RNAi-based control system against T. urticae.

We analyzed molecular and enzymatic properties of three cholinesterases (ChEs; ClAChE1, ClAChE2 and ClSChE) from Cimex lectularius. The ClAChE1 and ClAChE2 were generally present as a membrane-anchored dimeric insoluble form in the brain and ganglia. In the case of ClSChE, monomeric and dimeric soluble forms were observed. To investigate enzymatic properties, three ChEs were functionally expressed using baculovirus expression system. ClAChE1 revealed a significantly higher activity than ClAChE2 to acetylthiocholine iodide (ATChI) substrate. Kinetic analysis using two choline substrates (ATChI and butyrylthiocholine iodide) demonstrated that ClAChE2 had higher catalytic efficiency but lower substrate specificity than ClAChE1. Inhibition assay was conducted by using three inhibitors (BW284C51, eserine, Iso-OMPA) and two insecticides (chlorpyrifos-methyl and carbaryl). Two ClAChEs revealed high sensitivities to BW284C51, eserine, chlorpyrifos-methyl and carbaryl, but were not sensitive to Iso-OMPA. This inhibition profile confirmed that both ClAChEs are categorized as ChEs. Interestingly, the salivary specific cholinesterase did not show any measurable activities to choline substrates, confirming its non-synaptic function in C. lectularius

Metcalfa pruinosa (Say), native to North America, is spreading rapidly in the Korean Peninsula, causing serious damages on many deciduous forest trees, ornamental trees, and agricultural crops. Before the first report from Korea in 2005, M. pruinosa has not been reported from any other countries in the Eastern Palaearctic, while it has been record in Italy in 1979, and rapidly spreading into many European countries. To trace the invasion route of this species, we analyzed haplotype analysis of the mitochondrial cytochrome c oxidase subunit-I (mtCOI, 577bp), and developed microsatellite markers. In the haplotype analysis, a total of fourteen haplotypes were found from 69 individuals in 18 populations: 12 haplotypes (native region), 4 haplotypes (European region) and 2 haplotypes (Korea). Interestingly, Korean populations were clustered with some European populations. Eight polymorphic microsatellite loci were developed, and population structures were analyzed from 145 individuals in 8 populations. The origin and invasion route of M. pruinosa are under investigation.

In this study we apply Support Vector Machine (SVM) to the prediction of geo-effective halo coronal mass ejections (CMEs). The SVM, which is one of machine learning algorithms, is used for the purpose of classification and regression analysis. We use halo and partial halo CMEs from January 1996 to April 2010 in the SOHO/LASCO CME Catalog for training and prediction. And we also use their associated X-ray flare classes to identify front-side halo CMEs (stronger than B1 class), and the Dst index to determine geo-effective halo CMEs (stronger than -50 nT). The combinations of the speed and the angular width of CMEs, and their associated X-ray classes are used for input features of the SVM. We make an attempt to find the best model by using cross-validation which is processed by changing kernel functions of the SVM and their parameters. As a result we obtain statistical parameters for the best model by using the speed of CME and its associated X-ray flare class as input features of the SVM: Accuracy=0.66, PODy=0.76, PODn=0.49, FAR=0.72, Bias=1.06, CSI=0.59, TSS=0.25. The performance of the statistical parameters by applying the SVM is much better than those from the simple classifications based on constant classifiers.

In this paper, we describe the installation of VLBI Correlation Subsystem (VCS) main product and its performance at the Korea-Japan Correlation Center (KJCC). The VCS main product was installed at KJCC in August 2009. For the overall performance evaluation of VCS, playbacks, Raw VLBI Data Buffer (RVDB) system, and Data Archive (DA) system were installed together. The VCS main product was connected between RVDB and DA, and the correlation results were put into the DA to confirm the normal operation of VCS 16 station mode configuration. The evaluation test was first performed with 4 station mode, same as the factory test of VCS main product. Based on the results of 4 station mode, the same evaluation test was conducted for 16 station mode of VCS. We found that the correlation results of VCS were almost similarly compared to those of the Mitaka FX Correlator. Through the test results, we confirmed that the problems such as spectrum errors, delay parameter processing module and field programmable gate array errors in antenna unit, which were generated at the factory test of VCS main product, were clearly solved. And we verified the performance and connectivity of VCS by obtaining the expected correlation results and we also confirmed that the performance of VCS was sufficient for real VLBI observation data in both 4 and 16 station modes.

본 시험에서 건초(티머시, 알팔파 및 클라인)와 짚류(톨페스큐 및 볏짚)의 buffer 용해도와 단백질 분획이 실시되었으며, 조사료 자원의 buffer 추출이 in vitro 발효 성상, 분해율 및가스(CO2 및 CH4) 생성량에 미치는 효과를 조사하였다. 다른 조사료에 비해 총 단백질 중buffer 가용성 조단백질과 A fraction은 알팔파건초에서 각각 61% 및 41.77%로 가장 높았으며 볏짚에서 가장 낮았다(각각 42.8% 및19.78%). 총단백질 중 B1 fraction은 조사된 조사료간 비교적 큰 차이를 보이지 않았으나 B2fraction에서는 다른 조사료(6.34~8.85%)에 비하여 톨페스큐짚(10.05%) 및 클라인 건초(12.34)%에서 다소 높은 수준을 보였다. 총 단백질 중B3 fraction이 차지하는 비율은 톨페스큐짚에서38.49%로 가장 높았으나 다른 조사료 자원 간에는 큰 차이가 없었으며, C fraction의 경우 볏짚에서 가장 높은 비율(15.05%)을 보였다. 모든 사료에서 배양 개시 후 3시간(P<0.01) 및 6시간(P<0.05)에서 buffer 추출 전에 비해 추출후 배양액의 pH가 증가되었으며, 배양 6시간(P<0.05) 및 12시간(P<0.001)에서 다른 사료에비해 티모시 건초 및 알팔파 건초로부터의 pH가 낮았다. 배양액의 암모니아 농도는 모든 배양시간에서 가용성 물질의 추출 전 후에 다른조사료에 비해 알팔파 건초에서 가장 높았으나모든 사료의 추출효과는 배양 3시간(P<0.01)에서만 나타났다. 배양액의 총 VFA 농도는 배양24시간까지 알팔파 건초에서 가장 높았던 반면톨페스큐짚과 볏짚에서 가장 낮았다. 또한 모든 조사료에서 buffer 추출 전에 비하여 추출후에 총 VFA 농도가 감소되었다(P<0.01~P<0.001). Acetic acid (C2)의 조성 비율에서는 배양 6시간까지 추출 전에 더 높았으나(P<0.001)사료 간 차이는 없었다. Propionic acid (C3) 조성 비율 역시 배양 개시 후 3, 24 및 48시간(P<0.001)에 추출 전에 더 높았으며, 6 및 12시간의 배양액에서 대부분 건초(티모시, 알팔파 및 클라인)와 짚류(톨페스큐짚 및 볏짚) 간차이가 있는 것으로 조사되었다(P<0.05). 그러나 butyric acid (C4) 조성비율의 경우 대부분의배양시간에서 사료 간 차이는 없었다. 건물에서의 분해율 관련 parameter 중 a 값은 조사된전체 조사료에서 buffer 추출 전이 추출 후에비해서 높았으며(P<0.001), 다른 조사료에 비해톨페스큐짚과 볏짚에서 크게 낮았다(P<0.05).또한 b 값의 경우 역시 추출 전에 비해 추출후에서 현저히 낮았으나(P<0.001) 사료 간 차이는 없었다. 볏짚을 제외한 조사료에서 추출후에 비해 추출 전의 건물 유효분해율(EDDM)이 더 높았다(P<0.001). 조단백질에서의 a, b및 c 값은 추출 전에 비해 추출 후에서 현저히낮았으나(P<0.05) 사료 간 차이는 없었다. 조단백질 유효분해율(EDCP)에서는 다른 조사료 종류에 비해 톨페스큐짚과 볏짚에서 낮았다(P<0.05). 한편, NDF의 경우 a 값과 b 값(P<0.01)및 NDF 유효분해율(EDNDF, P<0.001)은 추출후에 비해 추출 전에 더 높았으나(P<0.01) 사료 간 차이는 보이지 않았다. 반추위미생물에의해 사료분해과정 중 생성되는 CO2 량도 24시간 배양까지는 추출 전에 더 많았으며(P<0.05~P<0.001), 톨페스큐짚과 볏짚에 비해 건초형태의 조사료로부터의 CO2 생성량이 더 많았다(P<0.05~P<0.01). 메탄가스(CH4) 생성량 역시 모든 배양시간에서 추출 전에 비해 추출 후에 크게 감소되었으며(P<0.01~P<0.001), 12~24시간을 제외하고는 짚류에 비해 건초에서 현저히 높은(P<0.05) 것으로 나타났다. 본 시험의 결과를 종합하면, 조사료 자원에 대한 buffer용해도와 단백질의 분획이 in vitro VFA 농도와 분해율 및 gas (CO2 및 CH4) 발생량 간 상호 밀접한 관계를 보이는 것으로 여겨진다. 이에 따라 조사료 이용 효율 개선을 위해 조사료자원에 대한 buffer 용해도와 단백질 분획을 반추동물 TMR 조제에 활용할 필요가 있는 것으로 여겨진다.

A study was conducted on four crossbred bulls, used as artificial insemination (AI) sires, to correlate their semen quality with their non return rate (NRR). Semen was collected once a week via an artificial vagina, diluted in egg yolk-citrate and maintained at for three days. It was evaluated for sperm motility, viability, morphology immediately after collection and was examined daily for sperm motility, viability and morphology of acrosome, mid piece and tail for a total of three days. A total of 2016 cows were inseminated by two AI technicians. The proportions of sperm with normal heads were 83.4% (63.7~91.7%), the proportion of spermatozoa exhibiting normal morphology (acrosome, mid piece and tail), motility and viability were 89.2% (82.3~92.0%), 71.3% (61.7~75.0%) and 76.7% (65.7~85.0%), respectively in fresh ejaculates. Sperm motility and sperm viability was significantly ( <0.05) lower in Holstein-Friesian Local bull than in other bulls during all three days of storage. The overall NRR for four bulls was 82.7% (72.9-87.5%). Bulls with higher sperm motility, viability and normal morphology of spermatozoa of individual bull had significantly (each <0.05) higher NRR. The highest ( <0.01) NRR (87.5%) was observed in a Red Chittagong bull whose semen qualities were significantly ( <0.05) higher than Holstein-Friesian Local bull (NNR 72.9%). The results of the present study concluded that NRR at 56 days post AI is related to parameters of semen quality. Therefore, semen evaluation may allow the discarding of bulls with poor fertility in an AI program.

Cytokines are known to function as regulatory molecules that can be produced by virtually every nucleated cell type in the body, including lymphocytes, monocytes/macrophages, epithelial cells, fibroblasts, and many others. Cytokines include lymphocyte-derived factors (lymphokines), monocyte-derived factors (monokines), hematopoietic factors (colony-stimulating factors), connective tissue/ growth factors, and chemotactic chemokines. Cytokines released in response to infection can affect tumor development in different ways. When exposed to infectious agents, cytokines are secreted by sentinel cells, such as macrophages and dendritic cells. These cytokines include interleukin 1 (IL-1) and tumor necrosis factor-α, as well as others, such as IL-6, IL-12, and IL-18. When released in sufficient quantities, these molecules can cause inflammation. Chronic inflammation is highly associated with tumor initiation, promotion, and progression. In this article, we review the roles and mechanisms of cytokines in tumor development.

Tungsten nanopowders were produced by the method of wires electrical explosion in the different gases. The study of phase and dispersed composition of the powders was carried out. The influence of electrical parameters such as the value of energy input in wire and the arc stage of the explosion was discussed. The factors that make for decreasing the particles size are the lower pressure of surrounding gas and the use of addition of chemically reactive gas.