To collect entomopathogenic fungi we sampled soils from two different cultural types of farm which are conducting conventional culture and organic culture, and non-crop land. Forty-seven soil samples were collected from 9 provinces. Among them, 7 samples were collected from non-crop land, 6 samples were from organic farm and 34 samples were from conventional culture farm. Insect-bait method using Galleria mellonella and agar plating method were conducted to collect entomopathogenic fungi. Collection of entomopathogenic fungi using insect-bait method was highest in non-crop land as 7.1 mycotic cadavers per soil sample followed by organic farm with 3.6 and conventional farm with 3.3. Result from agar plating method showed the highest colony forming unit (cfu) was in organic farm followed by non-crop land, but cfu in conventional cultivating farm was a half compared with the non-crop land.

Suspension culture is a useful tool for culturing embryonic stem (ES) cells in large-scale, but the stability of pluripotency and karyotype has to be maintained in vitro for clinical application. Therefore, we investigated whether the chromosomal abnormality of ES cells was induced in suspension culture or not. The ES cells were cultured in suspension as a form of aggregate with or without mouse embryonic fibroblasts (MEFs), and 0 or 1,000 U/ml leukemia inhibitory factor (LIF) was treated to suspended ES cells. After culturing ES cells in suspension, their karyotype, DNA content, and properties of pluripotency and differentiation were evaluated. As a result, the formation of tetraploid ES cell population was significantly increased in suspension culture in which ES cells were co-cultured with both MEFs and LIF. Tetraploid ES cell population was also generated when ES cells were cultured alone in suspension regardless of the existence of LIF. On the other hand, the formation of tetraploid ES cell population was not detected in LIF-free condition, in which MEFs were included. The origin of tetraploid ES cell population was turned out to be E14 ES cells and not MEFs by microsatellite analysis and the basic properties of them were still maintained despite ploidy-conversion to tetraploidy. Furthermore, we identified the ploidy shift from tetraploidy to near-triploidy as tetraploid ES cells were differentiated spontaneously. From these results, we demonstrated that suspension culture system could induce ploidy-conversion generating tetraploid ES cell population. Moreover, optimization of suspension culture system may make possible mass-production of ES cells.

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.

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.

Preservation of sperm is essential for long-term storage of valuable animal genetic resources and for the conservation of threatened mammalian species undergoing progressive extinction. In this study, using pig as a model system, we evaluated the feasibility of germ-plasm preservation via sperm cell lyophilization. We show that, pig sperm can be successfully lyophilized and stored in a liquid nitrogen-free condition for at least 6 months. Intracytoplasmic injection of lyophilized sperm (ICSI), stored at 4℃ for four months, into in vitro matured pig oocytes could successfully develop up to blastocyst stage (13.0±3.0%). Lyophilized sperm could also be stored at room temperature for at least three weeks without further compromising their in vitro development up to the blastocyst stage (14.6±3.2 vs. 16.6±5.1%; p>0.05). Blastocysts produced from ICSI of lyophilized sperm stored at 4℃ or room temperature contained similar number of cells per blastocyst (44.9±3.2 vs. 44.0±4.3; p>0.05) but was significantly lower than those produced from non-lyophilized fresh sperm (52.1±5.8 p>0.05). Interestingly, use of a custom-designed HEPES-buffered, calcium-free, defined medium for the lyophilization resulted in normal post-ICSI embryonic development up to blastula stage (23.4±2.8 vs. 24.0±2.9%) and, the resultant blastocysts contained similar number of cells per blastocyst (47.9±4.3 vs. 50.6±7.0) compared to those generated from non-lyophilized fresh sperm (p>0.05). These lyophilized sperm could also be stored at room temperature for at least three weeks with slight reduction in post-ICSI embryonic development (19.6±1.4%). Therefore, these results suggest that, pig sperm could be successfully and efficiently lyophilized for their long-term storage at 4℃. Lyophilization of sperm could be a practical option for long-term storage of mammalian germ-plasm.

Autophagy, the process of bulk degradation and recycling of long-lived proteins, macromolecular aggregates, and damaged intracellular organelles, has recently been shown to be important for pre-implantation development and cavitation in mouse embryos. This study investigated the occurrence of autophagy and its importance in determining the in vitro development of pig embryos produced by in vitro fertilization (IVF) or parthenogenetic activation (PA). Western blot analysis for autophagy marker, microtubule associated protein light chain 3 (MAP-LC3), revealed the temporal pattern of LC3-conversion with intense changes during 10 20 h post-insemination and at morula-blastocyst transition in pig embryos. Specific inhibition of autophagy in 2 4 cell stage pig embryos, by treatment with 3-methyladenine (3MA), did not affect their embryonic development up to morula stage (p>0.05) but completely blocked their progression to the blastocyst stage (0.0±0.0 vs. 28.5±1.7% p<0.05). On the other hand, autophagy-inhibition in morula stage embryos significantly inhibited the formation of blastocoel (14.9±3.6 vs. 37.5±7.2%) and reduced the proportion of expanded blastocysts (5.6±2.6 vs. 29.6± 4.6% p<0.05). TUNEL assay revealed that autophagy-inhibited embryos had significantly increased indices of apoptosis (10.2±0.4 vs. 2.3±0.2) and DNA fragmentation (0.8± 0.1 vs. 0.3±0.1) than those of controls (p<0.05). Interestingly, while anti-oxidants reduced (p<0.05) the apoptosis and improved the blastocyst formation rate in pig embryos, it had no influence (p>0.05) on the expression of MAP-LC3. These data therefore, suggest that autophagy may have essential role during blastocyst formation in pig embryos.

In the present study, total methanol extracts prepared from Alpinia katsumadai showed significant protective effects against the oxidative stress induced by hydrogen peroxide, UV-C or γ-ray irradiation. These protective effects were substantially increased by treatment with 20~100 μg/ml of the extract. The A. katsumadai total methanol preparation was further fractionated into n-hexane, dichloromethane, ethylacetate, n-butanol and water fractions. Among these five fractions, the ethylacetate and butanol fractions of A. katsumadai showed the strongest protective effects against oxidative stress induced by UV-C and γ-ray irradiation. These fractions also showed high DPPH radical scavenging and lipid peroxidation inhibitory activities. In addition, both fractions displayed cell proliferation activation effects, as evidenced by significant increases in colony formation. Our current data thus suggest that the mechanisms underlying the protective effects of A. katsumadai against oxidative damage may include radical scavenging, protection against cell membrane damage and stimulation of cell proliferation.

The purpose of this study is to investigate and reveal the effects that the complex exercise training consisting of aerobic exercise and strength training(sit up, push up) that everyone can easily practice regardless of a time and a place in order to manage practically the physical strength of the aged affects the difference on their body composition and the change of physical fitness level. Looking into the change of body composition of an experimental group, the weight of 2.5kg was reduced after applying complex training for 12 weeks and the body fat mass of 2.65kg was reduced. Also, the abdominal fat of 0.13% was decreased and the muscle mass of 1.56kg was increased. For the change factors of physical fitness, cardiovascular endurance, muscular strength, muscular endurance, balance and flexibility excluding agility showed significant improvement after applying complex exercise training. The improvement of health fitness of the aged under this study was significantly effective to improve specified body functions which had been lowered by aging and insufficient physical activities. So, it is regarded that their health fitness is the important factor to improve the activity competence required for daily life and to lead healthy living by the improved activity competence. Henceforth, it needs to study more the complex composition of several sports, exercise intensity and the frequency based on the previous researches and studies. In addition, it needs to develop the complex exercise training in accordance with various characteristics such as a sex of the aged, an age, a physical fitness level, environment, a disease and the program in consideration of the efficacy and safety during training.

In our present study, we investigated the effects of continentalic acid on Streptococcus mutans (S. mutans) biofilm. Methanol extract of Aralia continentalis (A. continentalis) was suspended in water and sequentially partitioned with CHCl3, ethyl acetate (EtOAc), and n-butanol (n-BuOH). The CHCl3 fraction showed the highest activity and an antibacterial compound against S. mutans was isolated from this preparation through various chromatography methods by bioassay guided fractionation. MS, 1H - NMR and 13C-NMR analysis showed that the active principle was continentalic acid which was confirmed to show significant inhibitory effects against S. mutans biofilm. These results may provide some scientific rationale for the traditional use these extracts for the treatment of dental diseases.

본 시험은 국화에서의 기형화 발생과 DNA 메틸레 이션에 관여하는 S-adenosyl-L-homocysteine hydrolase (SAHH) 유전자 발현과의 관련성을 검토하기 위해 수행 하였다. 스프레이 국화 ‘Lerbin’은 단일후 14일에서 27일 사이에 고온과 장일 조건에서 기형화가 발생되어, 35/20oC 의 고온과 14시간의 장일 처리 할 경우 25/20oC(12 h/ 12 h)에 비해 설상화가 2배 이상 증가하는 양상을 보였다. 스프레이 국화 ‘Lerbin’에서 분리한 전장 cDNA (DgSAHH) 는 크기가 1455 bp로 다른 작물과 90%이상의 높은 상동 성을 나타내었다. 국화의 DgSAHH 유전자 발현은 기형화 발생을 유도하는 고온과 장일 조건에서 크게 감소하였다. 이러한 결과를 통해 국화의 화아 발달에 있어서 DgSAHH 유전자 발현은 온도와 일장의 영향을 받으며, 억제된 이 유전자의 발현이 기형화 발생에 관여할 수 있을 것으로 판단된다.

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.

The purpose of this study was to identify the effects of manipulation on the velocity of cerebral blood flow and level of pain in cervicogeinc headache patients. The velocity of cerebral blood flow of 30 cervicogeinc headache patients(male=15, female=15, age=24.00±3.60) and 33 normal subjects(male=15, female=18, age=23.27±3.00) was compared. The 30 cervicogeinc headache patients were divided into suboccipitalis relaxation group, cervical manipulation group, and placebo group, and each were given different interventions. The velocity of cerebral blood flow and pain level was measured before intervention, and 1, 2, 3 weeks after intervention. The velocity of cerebral blood flow was measured with the Transcranial Doppler(TCD), and pain level was measured with visual analog scale(VAS). Blood flow velocity of middle cerebral artery in cervicogeinc headache patients was slower than those in healthy subjects. Physical therapy intervention did not have significant effect on velocity of cerebral blood flow, but slowly decreased at intervention for pain level increased. The suboccipitalis relaxation group and cervical manipulation group showed significant effect in decreasing pain level compared to the placebo group(p＜.05). Directly applied manipulation therapy in the neck area not only has effect on joint of cervical and soft tissue but also on blood vessels and nerves which pass the neck area, and because of those results of manual therapy seems to help recovery.

국화는 개화를 위해서는 30일의 지속적인 단일 처리가 요구되는 원예작물인데 본 연구에서는 국화의 화형별 홑꽃 타입의 ‘Limelight’, ‘Sunlight’, ‘Candle Light’, ‘Firebrand’, ‘Twilight’와 겹꽃 타입의 ‘Spirit’, ‘Sunburst Spirit’, ‘Mandalay’, ‘Illini Harvest’가 단일 (SD)−장일 (LD)−단일 (SD)의 일장 처리가 생장 및 개화조절 반응에 미치는 효과를 알아보고자 수행되었다. 30일간의 단일 처리와 5일 또는 10일 동안의 장일처리 후 5일 또는 10일 동안의 단일조건을 각각 처리하였다. 품종별 차이는 없었으나 5 SD−10 LD−25 SD 처리는 개화를 지연 시키고 설상화 수가 증가되었다. 특히 ‘Firebrand’와 ‘Candlelight’ 품종의 설상화 수는 각각 40개와 60.8개로 대조구에 비해 현저히 증가되었다. 또한 SD−LD−SD 처리는 개화를 6일에서 7일정도 지연시키는 것 외에는 큰 효과를 나타나지 않았다. 그러나 ‘Illini Harvest’와 ‘Limelight’의 설상화 수는 어떤 SD−LD−SD 처리에 의해서도 증가되지 않았다. 따라서 추후 새로운 품종별 일장처리에 따른 추가적인 연구 분석이 뒤따라야 할 것으로 판단되며 본 연구 결과는 국화의 신품종 육성을 위한 자료에 유용함을 알 수 있었다.