True hermaphrodites are animals of equivocal sex in which both male and female gonads develop simultaneously. The frequency of true hermaphroditism is higher in pigs than in other domestic animals. Two Korean pigs were diagnosed with true hermaphroditism showing ovotestes, epididymes, penes, and uteri. Histomorphologically, the testicular tissues consisted of Sertoli cells that were devoid of spermatogenic germ cells and showed proliferation of interstitial cells. However, the uteri were of normal architecture and had well-developed uterine endometrial glands. The samples were 38, XX female karyotype without the sex-determining region Y (SRY) gene. The findings of this study could contribute to the understanding of true hermaphroditism in the Korean pig industry. * This work was supported by a grant (Code# PJ008148) from BioGreen21 Program, Rural Development Administration, Republic of Korea.

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 purpose of this study was to examine the degree of knowledge of care workers working at long term care hospitals and nursing homes on pressure ulcer. A total of 81 care workers including 34 at long term care hospitals and 47 at nursing homes were surveyed. 24 questions were used to evaluate their degree of knowledge on pressure ulcer. Their knowledge on pressure ulcer scored 12.84 out of the total score of 24 points(SD=3.40), which was equal to 53.50 (SD=24.23) out of 100 points. Their knowledge on the prevention of pressure ulcer was highest among the subareas of evaluation. Their knowledge on pressure ulcer statistically significantly differed according to education on pressure ulcer(P<.05). A pressure ulcer is a skin disorder that may be prevented and cured. At this point when long term care facilities are rapidly increasing, care workers highlevel knowledge on and good management of pressure ulcer is very important. Practically educating them on pressure ulcer including the provision of recent, updated relevant knowledge will be necessary.

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.

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.

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.

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%).

To find an alternative for synthetic pesticides, methanol extract from plant samples were tested for their insecticidal activity against insect. The extract of Asiasarum sieboldii had strongly insecticidal activity against Plutella xylostella. Roots of A. sieboldii were extracted with methanol, and the concentrated extract was partitioned with n-hexane, ethylacetate, n-buthanol and H2O. The highest activity was shown in the hexane fraction. Activity-guided fractionation led to the isolation of two amides from hexane fration through the repeated silica gel column chromatographic separations. From the interpretation of spectropic data including NMR, MS, IR, the chemical structures of compounds were determined as dodeca-2E,4E,8Z,10Z-tetraenoic acid isobutylamide and dodeca-2E,4E,8Z, 10E-tetraenoic acid isobutylamide. These compounds showed insecticidal activity on P. xylostella by 96.7% at 100ppm. The liquid formulation controlled on cabbage effectively. The extract and compounds from A. sieboldii showed insecticidal activity against Nilaparvata lugens. As a naturally occurring pesticide, A. sieboldii could be useful as a new botanic insecticide.

Recently, farmers of mushroom farms in Korea express the difficulty of shiitake cultivation due to the damage of oak log beds by unknown reason. To elucidate the cause of the damage, we investigated and isolated fungi from inside wood chip of the damaged shiitake log bed. Four of green fungi were obtained and their morphology and molecular properties examined. Trichoderma atroviride, Trichoderma citrinoviride, Hypocrea lixii (Trichoderma harzianum), and Hypocrea lutea (Gliocladiumviride) were identified. Hypocarea lutea is newly described in Korea. These species were able to produce extracellular enzymes such as cellulase, pectinase, and xylanse that might degrade wood components of oak logs. This study provides strong evidence that Trichoderma fungi are involved in the recent damages of shiitake log beds In Korea.

The purpose of this study was to analyze and compare the effect of resistance exercise and balance exercise on proprioception and WOMAC index of patients with degenerative knee osteoarthritis. A total of 40 subjects participated in this study. The subjects were diagnosed with degenerative knee osteoarthritis and all were more than 60 years old. They were divided into three groups. Group Ⅰ(n=8) was trained with resistance exercise, Group Ⅱ(n=6) was trained with balance exercise and GroupⅢ(n=6) was trained with range of motion as a control. The results of this study were as follows. It was significantly indicated that the resistance exercise group and balance exercise group elicited error-reduction on proprioception goal-angle (p<.05). There was a statistically significant difference on proprioception between resistance exercise group and control(range of motion) group. There was a statistically significant reduction on WOMAC index between resistance exercise group and balance exercise group (p<.05) and on the WOMAC index between resistance exercise group and range of motion group(p<.05). In conclusion, resistance exercise and balance exercise are effective on degenerative knee osteoarthritis and resistance exercise is the most effective for improving proprioception and WOMAC index. More research on the intervention according to the degree of degenerative knee osteoarthritis is needed.

The effects of chitosan upon the experimentally induced differentiation of MDPC-23 cells, derived from mouse dental papilla cells, were investigated by RT-PCR, observations of cell morphology and Alizaline red-S staining. Chitosan was found to significantly increase and accelerate the expression of ALP mRNA but decrease the ColI transcript levels, as compared with the control, in a time-dependent manner during the differentiation of MDPC-23 cells. Chitosan also significantly downregulated ON mRNA expression and accelerated mineralization in differentiating MDPC-23 cells. These results suggest that chitosan facilitates odontoblast differentiation and mineralization and may have potential clinical applications as a dentin regeneration material.

The objective of this study was to develop a reliable and reproducible descriptive analysis procedure for Korean style sweet pumpkin gruel (Hobakjuk). The sensory attributes of the sweet pumpkin gruel were developed and defined, the sample preparation method was standardized， and the sensory evaluation procedure for a sample was established. Seven types of sweet pumpkin gruel (five ready-to-eat type vs. two ready-to-heat type) were selected to be analyzed. Panel training and descriptive analysis were carried out with these 7 samples. A total of 12 sensory attributes (2 aroma/odor, 5 taste/flavor, 4 texture/mouthfeel, and 1 aftertaste attributes) were developed to describe the sensory characteristics of the sweet pumpkin gruel. The definition and reference standards for each sensory attribute were determined to clearly understand each attribute. In the main experiment, trained panelists evaluated the sensory characteristics of the 7 gruel samples based on a fifteen-point intensity scale using the developed attributes. The results were statistically analyzed by analysis of variance, principal component analysis, and cluster analysis. The results showed that the 7 sweet pumpkin gruel samples significantly diffieredin their intensities of all attributes except for sweet pumpkin aroma and viscosity. The ready-to-eat style samples were distinctly characterized by their sweet pumpkin aroma and flavor, whereas the ready-to-heat style samples were markedly characterized by their low intensity of gelatinized starch and pumpkin flavor retention.

We determined the complete mitochondrial genome (mitogenome) of the Japanese Oak Silkmoth, Antheraea yamamai (Lepidoptera: Saturniidae) from two overlapping fragments and subsequent shotgun sequencing. The 15,601-bp long A. yamamai mitogenome contains gene arrangement and content identical to the most common arrangement found in lepidopteran insects. Most individual A. yamamai mitochondrial (mt) genes were well within the range found in the respective genes of other insects, except for small ribosomal RNA (1,037 bp). The 336-bp A+T-rich region is relatively smaller than that of other lepidopteran insects. The region is interesting in that it contains tRNA-like structures as found in the A+T-rich regions of other insect mitogenomes. The start codon of A. yamamai COI gene is unusual in that no typical one (ATN) is available. Three of the 13 protein-coding genes have incomplete termination codon T or TA. All tRNA formed stable stem-and-loop structure, except for tRNASer(AGN), the DHU arm of which formed a simple loop as seen in many other metazoan mt tRNASer(AGN).