The aim of this study was to examine the effect of acteoside (the cyclin-dependent kinase inhibitor) on the SCNT efficiency with adult fibroblasts in dog. Canine adult fibroblasts were obtained from muscle and cell cycle of fibroblasts was synchronized by culturing to confluency, serum starvation and treating with 30 μM acteoside for 48 h. Cell cycle stages, cell cytotoxicity (apoptosis) and, prduction of reactive oxygen species (ROS) were analyzed using flow cytometry. The canine cells, prepared by confluent-cell culture or treating with 30 μM acteoside for 48 h, were injected into enucleated in vivo matured oocytes, the couplets were electrical fused and activated by calcium ionomycin. SCNT embryos using acteoside-treated fibroblasts were surgically transferred into oviducts of estrus cycle synchronized recipient dogs. In cell cycle synchronization (G0/G1), there was no significant difference between serum starvations (83.9%) and acteoside treated groups (81.3%) that were higher than confluent group (78.5%). In production of apoptosis, confluent and acteoside treated groups (4.3 and 4.5%, respectively) were generated less than serum starvation group (21.8%). In case of ROS, serum starvation group was induced a significantly higher than other groups. After synchronization of the donor cell cycle, either confluent or acteoside treated, cells were placed with enucleated in vivo-matured dog oocytes, fused by electric stimulation, activated, and transferred into naturally estrus-synchronized surrogates. Fusion and cleavage rate of acteoside treated group were 64.1 and 41.5%, which were higher than those of confluent group (53.9 and 20.6%, respectively). The reconstructed embryo development rates to 4-cell and 8-cell in acteoside treated group were 29.5 and 14.8%, respectively, while confluent group showed 11.1 and 3.2%, respectively. Total 54 SCNT embryos using acteoside-treated fibroblasts were transferred into oviducts of 2 recipient dogs and one recipient finally delivered one puppy, whereas din`t detected pregnancy on transfer of cloned embryos reconstructed with confluent cells in 6 surrogate dogs. In conclusion, the results of the current study demonstrated that canine fibroblasts could be successfully arrested at the G0/G1 stage with reduced the formation of ROS and apoptosis after acteoside treatment. This results may contribute to improve the effi-ciency of canine SCNT. * This research was supported by iPET (Grants 110056-3), Ministry for Food, Agriculture, Forestry and Fisheries, Republic of Korea.

Acteoside acts as an anti-oxidative activity and anti-apoptosis in the cells. But, it has been not studied on maturation and development of porcine oocytes. The aims of the present study were to examine the effects of acteoside on the morphological progress of meiosis, developmental competence, and ROS in porcine oocytes. Oocytes were matured in tissue culture medium-199, supplemented with acteoside at various concentrations: 0 (control), 10, 30 and 50 μM. The oocytes maturation rates of groups supplemented with acteoside were no significantly different (81.13, 85.96, 82.95 and 83.68%, respectively). Level of ROS was significantly decreased in acteoside treated group. Furthermore, the parthenogenetic blastocyst rate was significantly improved in 10 μM acteoside treated group compared with control group (44.83 vs. 27.75%). And we investigated effect of acteoside on the oocytes condition represented by cytoplasmic maturation by homogeneous distribution and formation of cytoplasmic organelles and regulation of apoptosis-related genes. In the results. during IVM, 10 μM acteoside treated oocytes showed that the mitochondria and lipid droplet were smaller and homogeneous distribution in cytoplasm compare with control oocytes. And reverse transcription polymerase chain reaction (RTPCR) of parthenogenetic blstocysts revealed that acteoside increased the anti-apoptotic genes (Mcl-1, Bcl-2 and Bcl-xL), whereas reduced the expression of pro-apoptotic genes (Bax and Bak). In conclusion, based on the results, the effect of acteoside on IVM was not attractive. However, in acteoside treated group, cytoplasmic maturation seemed to be improved with morphologically uniform distribution of cytoplasmic organelles. Furthermore, embryonic development in acteoside treated group was significantly highly increased than that of non-treated group. Our results represents that addition of acteoside to the IVM medium has a beneficial effect in physiology of porcine oocytes, providing a improved method for porcine oocytes in vitro. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

The objective of the current study was to describe in vitro embryo production in Hanwoo, analyzing oocytes yield and embryo production. The effects of oocytes production and the number of OPU procedures per animal on embryo production were also evaluated. OPU was done every 3～4 days during experimental period and collected oocytes were fertilized in vitro in both OPU and needle puncture groups. First, we compared the recovery rate of oocytes based on OPU session (Experiment 1). The average of collected oocytes was calculated from every 10 session. The average number of total oocytes recovered per animalonsessionwas 5.16 (mean). Second, we compared the recovery rate base on collection period of OPU (Experiment 2). The following results show the difference of the number of recovered oocytes in every month during the procedure between the months of session. Every animal shows the constant number of recovered oocytes for the first 5 months. However, the recovery rate of oocytes was decreased from month 6 to 8. Third, we compared the developmental rate to blastocyst in two groups (Experiment 3). Oocytes by needle puncture were fertilized with frozen-thawing semen; the cleavage rate 24～48 h after in vitro fertilization (IVF) was 75.8% and blastocyst development rate was 18.8% in needle puncture group. Even though there is lower cleavage rate after IVF in OPU group (61.1%), blastocyst development rate was higher compared with needle puncture group (28.4%). 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.

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.

운간초의 분화 품질을 향상시키기 위하여 ‘Kumoma’ 와 ‘Kumoma-Gusa’ 두 품종의 생장과 개화에 미치는 영향을 구명하고자 식물생장억제제 (PGRs) 4종을 처리 하였다. 식물생장억제제 종류와 처리농도는 paclobutrazol (10, 20, 40, 80 mg·L-1), flurprimidol (5, 10, 20, 40mg·L-1), daminozide (500, 1000, 2000, 4000mg·L-1), chlormequat (50, 100, 200, 400 mg·L-1)이고 엽면살포 와 토양관주 방법으로 처리하였다. Paclobutrazol 40 mg·L-1 처리는 운간초의 두 품종에서 초장과 화경 장을 줄이는데 효과적이었다. ‘Kumoma’ 품종에서, paclobutrazol 40 mg·L-1 엽면살포와 토양관주 처리방 법은 초장을 각각 12.6, 12.5 cm로, 화경장은 3.4, 3.3 cm로 줄일 수 있었다. ‘Kumoma-Gusa‘ 품종에서 는 paclobutrazol 토양관주는 처리 농도가 증가함에 따라 초장은 13.2-10.7 cm, 화경장은 3.9-2.0 cm까지 줄일 수 있었지만 꽃의 수는 대조구와 비교했을 때 현 저하게 감소하여서 20개까지 감소하였다. Flurprimidol 20mg·L-1 스프레이와 토양관주 처리는 두 품종 모두 초장과 화경장을 줄일 수 있었다. ‘Kumoma’ 품종에 서 flurprimidol 10 mg·L-1 스프레이 또는 토양관주 처리는 개화 품질을 향상시킬 수 있었고 대조구와 비 교해 꽃의 수는 44.7개 이상, 출하 일수는 5일 정도 빨리 출하 할 수 있었다. ‘Kumoma-Gusa‘ 품종에서 PGRs처리에 의한 초장 과 화경장 억제효과가 가장 큰 것은 paclobutrazol 80mg·L-1과 flurprimidol 40mg·L-1 이었고 paclobutrazol 과 flurprimidol 스프레이와 토양관주 처리에서 각각, 초장은 10.7, 9.9 cm, 화경장은 2.0, 1.5 cm 로 생장 억제효과가 가장 큰 것을 알 수 있었다. 그러나 flurprimidol 40 mg·L-1 토양관주 처리에서 출하일수는 농도에 따라 3-13일 정도 대조구보다 늦어졌고 꽃의 수는 15.6개로 가장 적은 것을 관찰 할 수 있었다. 두 품종 모두에서 daminozide 처리는 초장과 화경장 이 줄어들지 않았지만 ‘Kumoma’ 품종에서는 대조구 와 비교했을 때 꽃의 수가 증가하였다. Chlormequat 처리는 paclobutrazol 과 flurprimidol 처리와 비교했 을 때 화경장 감소효과는 작았고 처리 식물의 잎은 다른 처리에 비해 연한 녹색을 보여주었다. 두 품종 모두에서 chlormequat 과 daminozide 처리는 생육과 개화 품질에 효과적으로 영향을 주지는 않았지만 꽃의 수는 모든 처리농도에서 41개 이상을 보여주었고, ‘Kumoma-Gusa’ 품종에 chlormequat 토양관주 처리는 최대 꽃의 수 63개를 볼 수 있었다. 이번 결과들은 paclobutrazol 과 flurprimidol 처리가 운간초 분화의 품질 향상을 위한 화경장의 생육을 조 절하는 생장억제제로서 사용 될 수 있음을 보여 주었 고, ‘Kumoma’ 품종에서는 paclobutrazol 40 mg·L-1 스프레이는 생육특성을, flurprimidol 10 mg·L-1 토양관 주 방법은 개화관련 특성의 품질을 향상시키는 생장억 제제의 농도로 추천 될 수 있고, ‘Kumoma-Gusa’ 품종 에서는 flurprimidol 20mg·L-1 스프레이와 flurprimidol 5mg·L-1 토양관주 방법이 운간초 분화의 생육특성과 개 화품질을 향상시키는 생장억제제의 농도로 추천 될 수 있다.

Two-spotted spider mite, Tetranychus urticae (Koch) is one of the serious pests in economically important crops such as strawberry and cucumber and so on. Acaricides have been used as the main control agents. This study was conducted to test the synergistic effects of the Beauveria bassiana GHA, which has been registered for whitefly and thrips, and five pesticides (abamectin, acrinathrin, bifenthrin imidacloprid, dinotefuran, and indoxacarb, which are commonly used pesticides in strawberry in Korea), on the two-spotted spider mite T. urticae. Five tested pesticides did not inhibit spore germination and mycelial growth of B. bassiana. Pesticides were applied to potted strawberry plants at four different treatments (recommended concentration, 1/5 recommended concentration, 1/5 recommended concentration + GHA(108 conidia/ml), and only GHA(108 conidia/ml). Mortality in larvae of two-spotted spider mite was 12% in GHA 5 day after treatment. Mortality in abamectin treated T. urticae was 98.5% and 100% at 1/5 recommended concentration and recommended concentration, respectively, 3 days after treatment. Acrinathrin, indoxacarbe, dinotefuran and difenthrin imidacloprid caused 60%, 14%, 16% and 91% mortality at recommended concentration 5 day after treatment. The tested five pesticides and B. bassiana GHA had no synergistic effect.

에멀션 기반의 계면활성제를 이용한 주형합성법을 이용하여 산촉매로서 염산과 실리카의 전구체인 테 트라에톡시실란을 사용함으로써 메조다공성 실리카 마이크로스피어를 합성하였다. 테트라에톡시실란의 농도 증가에 의해 구형의 입자 형태가 파괴되었고, 기공구조도 크게 변하였다. 산촉매 농도 증가에 의한 구형의 입자형태 파괴 현상은 적었지만 상대적으로 작은 크기의 구형의 마이크로입자가 더 많이 생성되 었다. 하지만, 산성조건에서 입자들 간의 강한 응집현상이 나타남에 따라 낱개의 분리되어 있는 단일입자 를 얻기 위해서는 초음파 등의 후처리 과정이 필요하였다.

We analyzed the chirp sound and behavior of Teleogryllus emma with observation system, which was consisted of computer, ccd-camera and microphone. Computational methods of wavelet transformation and Self-Organizing Maps (SOM) were utilized to characterized the chirp sound of insect species for automatic counting in this study. Wavelets were initially applied to feature extraction of the chirp sound. Wavelet coefficients were accordingly calculated based on the basis function (e.g., Morlet). The obtained coefficients were subsequently provided to count number of chirps in each song. Sound structure of insect specimens consisted with long chirp and short chirp and the patterns of song were grouped by frequency of long chirp and short chirp. The song patterns of insect specimens were divided by Self-Organizing Map (SOM) that was used number of chirp as input data. Application of computational methods to automatic detection of chirp sound was further discussed for obtaining objective assessment in behavior science.

수평 배향 액정 모드는 양의 유전율 이방성과 음의 유전융 이방성 액정을 사용한 프린지 필드 스위칭과 양의 유전율 이방성 액정을 사용한 인플래인 스위칭 모드가 대표적이다 . 이 대표적인 세 구동 방식의 화질 특성을 비교하기 위하여 각각의 최척화된 위상지연 값 조건하에서 밝기, 명암대비율과 색 특성을 조사하였다. 그 결과 양액정과 음액정을 사용한 프린지 필드 스위칭 모드가 밝기와 명암대비율 면에 있어서 인플래인 스위칭 모드보다 우수한 특성을 보인다. 또한 양액정을 사용한 프린지 필드 스위 칭 모드는 시야각 방향에서 적은 색 변이 특성을 보인다.

The neurotrophin plays an important role in the development, differentiation and survival of the nervous system in vertebrates. It exerts its cellular effects through two different receptors, the Trk receptor tyrosine kinase neurotrophin receptor and the p75 neurotrophin receptor, a member of the tumor necrosis factor receptor superfamily. Trk and p75 neurotrophin receptors utilize specific target proteins to transmit signals into the cell. An ankyrin-rich membrane spanning protein (ARMS) was identified as a new p75 interacting protein and serves as a novel downstream target of p75 neurotrophin receptor. We sought to delineate the interaction between p75 and ARMS by deletion constructs of p75 and green fluorescent protein (GFP)-tagged ARMS. We examined the interaction between these two proteins after overexpressing them in HEK-293 cells. Using both Western blot analysis and immunocytochemistry followed by confocal laser scanning microscopy, we found out that the intracellular domain of the p75 neurotrophin receptor was important for the interaction with ARMS. The results from this study suggest that ARMS may play an important role for mediating the signals from p75 neurotrophin receptor into the cell.

Field sequential 액정 디스플레이(FSLCD)는 컬러필터를 사용하지 않아 높은 투과율 특성을 보이고 광 원으로 LED를 사용함으로써 색재현성이 매우 우수하다. 하지만 FSLCD(60Hz 구동)를 실현하기 위해서는 액정의 응답속도가 5ms이하로 고속응답 특성을 보여야 한다. 따라서 본 논문에서는 고속응답 ECB(electrically controlled birefringence) 셀의 최적 구조를 연구하여 5ms 이하의 응답시간을 얻었다. 그리고 ECB 모드에서 높은 구동전압과 시야각을 개선하기 위해 필름 보상을 연구하였다. 판상형 액정필름(discotic film)과 TAC(triacetyl cellulose) 필름의 위상차 값을 최적화함으로써 구동전압을 5V로 낮추고 상하좌우에서 160° 이상(CR>10:1)의 시야각을 실현하였다.

The purpose 01' pl'esent study was to examine the molecular events in apoptosis by CoCl2, mimicking hypoxic cond ition and recovering effects by LED ir l'adiation on Human SH-SY5Y neuroblastoma cells The SOUl'ce 0 1' light for ir l'adiation was a continuous-wave LED emitting at a wavelenl양h of 590 nm, and manufactured that ene rgy density was 5 mW!cm2 on sample surface, After ir l'adiation, cell viabi lity was measured with BrdU , cell morphol ogy was examined with Diff- Quik staining, cell signaling was monitored with various apoptosis-related molecules using RNase Pl'otection Assay(RPA) , W11en treated with CoC12, apoptotic induction was found in the SH-SY5Y cells in a concentration-dependent and time-dependent manner , Diff-Quik s taining was revealed that DNA fragmentation re presented apoptosis was examined in CoC12-tl'eated group, Moreover, RPA assay of SH-SY5Y cclls lIs ing val'iolls apoptosis-related molecllles showed that the apoptotic cell population was mcreased J-loweve. there was sorne signifïcant change in LED irradiatied cells aftel' treatement of CoC12 The main mechanism for Lhese a poptosis appearecl to be mito c hondriεt - m ecliated pathway, such as cytochrome- c‘ caspase-9, caspase-3, pro-apototic protein ßax, anti-apototic protein Bcl-2, and death receptor• mediated pathway, such as Fas, cas pase- 8, a ncl TNFRl These results demonstrate that CoCI2 induce apoptosis in SH-SY5Y via different dual apop tosis pathway through death receptor pathway as well as mitochondria- dependent pathway and LED irradiation can recl llces the CoCl2-induced apoptosis by blocking their internal signaling pathway

Mushrooms have been widely cultivated and consumed as foods and herbal medicines owing to their various biological properties. However, few studies have evaluated the anti-inflammatory effects of mushrooms. Here, we investigated the effects of mushroom extracts (MEs) on lipopolysaccharide (LPS)-induced inflammation in macrophages (RAW264.7 cells). First, we extracted MEs with either water or ethanol. Using LPS-treated RAW264.7 cells, we measured cell proliferation and NO production. Gene expression of tumor necrosis factor-α (TNF-α), interleukin (IL)-6 (IL-6), and IL-1β was assessed by RT-PCR, and protein abundance of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) and phosphorylation of p65 were determined by immunoblotting. MEs prepared using both water and ethanol inhibited LPS-induced inflammation in RAW264.7 cells. Nitric oxide (NO) levels induced by LPS were reduced by treatment with MEs. Isaria japonica Yasuda water extracts and Umbilicaria esculenta (Miyoshi) Minks ethanol extracts significantly decreased the mRNA expression of inflammation-related cytokine genes including TNF-α, IL-6, and IL-1β. Similarly, the protein abundance of iNOS and COX-2 was also decreased. The phosphorylation of p65, a subunit of nuclear factor-κB was at least partly suppressed by MEs. This study suggests that mushrooms could be included in the diet to prevent and treat macrophage-related chronic immune diseases.

Background : Cordyceps militaris is a non-toxic, medicinal mushroom, which is known to possess anti-inflammatory and immunomodulating activities. And also, Glycyrrhiza uralensis is widely used as a crude drug in oriental medicine. However, the effects and mechanism of action of C. militaris and G. uralensis on Herpes simplex virsu (HSV), which is a serious skin disease. This study aimed to evaluate the effect of C. militaris and G. uralensis on Herpes simplex virus. Methods and Results : The results showed that the extracts and major compounds C. militaris and G. uralensis increased the TNF-α product on RAW 264.7. And also, these extracts and major compounds inhibited TNF-α product in RAW 264.7 induced by LPS. Querceitn, which was identified from G. uralensis, was showed Anti-virrus effect of Herpes simplex virus (HSV). Conclusion : Taken together, these results indicate that the anti-stomach cancer effect of C. militaris and G. uralensis in xenograft model implantated Epstein-Barr virus positive-stomach cancer cell line.

Background : Cordyceps militaris has been an wonder drug to anti-aging efficacy and called the three main drugs with ginseng and deer antler from the past. Cordycepin, cordycepic acid (d-mannitol) and adenosine are known as functional ingredients in Cordyceps militaris. Among them, cordycepin, the representative component, has been reported as antimicrobial substance containing immune enhancement, anti-cancer and anti-inflammatory effects. Methods and Results : After Cordyceps militaris produced from different types of medium mixed with 10-fold volume of purified water, the mixture were extracted at 70±5℃ for a hour and that extracts re-extracted using ultrasonics wave for 30 minutes. Qualitative analysis of the index component was determined by using the Q-TOF (A quadrupole time-of-flight mass spectrometer), and quantitative analysis was performed by using HPLC (high-performance liquid chromatography) with Xselect HSS T3 column (2.1 X 100 mm, 2.5㎛, Waters, USA) and ultrapure water and acetonitrile as mobile phase A and B. Detection column temperature, injection volume and the flow rate were 35℃, 2 μL and 0.3 mL / minute respectively. The cordycepin content of Cordyceps militaris produced from medium mixed with vegetable and animal ingredients higher than single ingredient. Moreover, through a variety of analyzes by varying the type and content of the medium additives, the cordycepin in Cordyceps militaris produced from medium mixed with animal ingredients highest. Furthermore, the cordycepin content of a fruit body was higher than those of the a mycelium. Conclusion : These results provide a method for producing an high cordycepin content of Cordyceps militaris as functional food ingredient.

Acute myocardial infarction (AMI) involving two simultaneous vascular territories of acute transmural ischemia is known as a double or combined myocardial infarction. This well described but extremely rare phenomenon may be related to the fact that AMI with multiple vessel obstruction often causes extensive myocardial injury and death before hospitalization. We reported the first case of 45-year-old man with AMI occluded left anterior descending artery and right coronary artery simultaneously in Korea.