Entomopathogenic fungi are natural enemies of insect pests and contribute to the natural regulation of their host populations. These fungal group are often used as active ingredients for microbial insect pest control. In addition, the potential antimicrobial effect by entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have recently been reported against fungal plant pathogens. Dual microbial control effects with entomopathogenic fungi against both aphids and cucumber powdery mildew had reported in Canada. In our previous studies we conducted bioassay with entomopathogenic fungi to develop dual microbial control agent which can control both aphid and fungal plant disease. We selected an Beauveria bassiana isolate which has high dual control effects against both cotton aphid, Aphis gossypii and sclerotinia rot, Sclerotinia sclerotiorum. In this study, we have tested the dual control efficacy of the B. bassiana isolate against cotton aphid and sclerotinia rot on whole potted cucumber plants. We found that the B. bassiana isolate protected the plant from cotton aphid and sclerotinia rot under laboratory condition.

Acetylcholinesterase (AChE) is a hydrolase that hydrolyzes the neurotransmitter acetylcholine. Soluble form of AChE is generated via alternative splicing and functions as a bioscavenger in Dropsophila melanogaster. In this study, effects of acetic acid on the soluble AChE expression were investigated. Treatment of acetic acid resulted in over-expression of soluble AChE in the abdomen in a dose-dependent manner. The soluble AChE was determined to be expressed in the fat body. However, no apparent change in AChE expression was observed in the head. Our finding suggests that the soluble AChE is involved in chemical defense against high concentration of acetic acid, which is a common by-product in fermenting foods. The high level of acetic acid resistance in D. melanogaster, thus, appears to have been evolved via the induction mechanism of soluble AChE expression.

Despite many researches related with in-vitro culture of porcine spematogonial stem cells (SSCs), adherent culture system widely used has shown a limitation in the maintenance of porcine SSC self-renewal. Therefore, in order to overcome this obstacle, suspension culture, which is known to have numerous advantage over adherent culture, was applied to the culture of porcine SSCs. Porcine SSCs retrieved from neonatal testes were suspension-cultured for 5 days or 20 days, and characteristics of suspension-cultured porcine SSCs including proliferation, alkaline phosphatase (AP) activity, and self-renewal-specific gene expression were investigated and compared with those of adherent-cul-tured porcine SSCs. As the results, the suspension-cultured porcine SSCs showed entirely non-proliferative and significantly higher rate of AP-positive cells and expression of self-renewal-specific genes than the adherent-cultured porcine SSCs. In addition, long-term culture of porcine SSCs in suspension condition induced significant decrease in the yield of AP staining-positive cells on post-day 10 of culture. These results showed that suspension culture was inappropriate to culture porcine SSCs, because the culture of porcine SSCs in suspension condition didn’t stimulate proliferation and maintain AP activity of porcine SSCs, regardless of culture periods.

The physiology of parasitic wasp control of their lepidopteran hosts' not only includes injecting their egg but also various factors such as symbiotic virus. This study was focused on the investigation of sophisticated interaction between parasitoid (Diadegma fenestrale) and their host (Plutella xylostella) in P. xylostella larva at transcriptome level, to check whether it is parasitized or not. Short-read deep sequencing method (Hiseq2000) was used for the transcriptome analysis. De novo assembly of cDNA sequence data generated 196,081 contigs between 201bp and 15,853bp in length. Some detoxification enzymes such as cytochrome P450 and Immune-related genes such as antimicrobial peptides were up-regulated after parasitism. Expression of symbiotic ichnovirus genes was detected in parasitized larvae with 55 contigs identified from five ichnovirus gene families including vankyrin, viral innexin, repeat elements, a cysteine-rich motif, and polar residue rich protein. This investigation provides a detailed information on differential expression of P. xylostella larval genes and symbiotic ichnovirus genes following parasitization.

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.

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.

Several lines of evidence suggest that osteocytes play a critical role in bone remodeling. Both healthy and apoptotic osteocytes can send signals to other bone surface cells such as osteoblasts, osteoclasts, osteoclast precursors, and bone lining cells through canalicular networks. Osteocytes responding to mechanical strain may also send signals to other cells. To determine the role for osteocytes an mechanical strain in bone remodeling, we examined the effects of fluid flow shear stress on osteoclast precursor cell and osteoblast proliferation and recruitment induced by osteocytes. In addition, the effects of fluid flow shear stress on osteocyte M-CSF, RANKL, and OPG mRNA expression were also examined. MLO-Y4 cells were used as an in vitro model for osteocytes, RAW 264.7 cells and MOCP-5 cells as osteoclast precursors, and 2T3 cells as osteoblasts. MLO-Y4 cells conditioned medium (Y4-CM) was collected after 24h culture. For fluid flow experiments, MLO-Y4 cells were exposed to 2h of pulsatile fluid flow (PFF) at 2, 4, 8, 16±0.6dynes/cm² using the Flexcell StreamerTM system. For proliferation assays, MOCP-5, RAW 264.7, and 2T3 cells were cultured with control media or 10-100% Y4 CM. Cells were cultured for 3d, and then cells were counted. RAW 264.7 and 2T3 cell migration was assayed using transwells with control media or 10-100% Y4-CM. M-CSF, RANKL and OPG in MLO-Y4 mRNA expression was determined by semiquantitative RT-PCR. Y4-CM increased osteoclast precursor proliferation and migration, but decreased 2T3 cell proliferation and migration. CM from MLO-Y4 cells exposed to PFF caused decreased RAW 267.4 cell proliferation and migration and 2T3 migration compared to control Y4-CM. However, Y4-CM from cells exposed to PFF had no effect on 2T3 osteoblastic cell proliferation. PFF decreased RNAKL mRNA and increased OPG mRNA in MLO-Y4 cells compared to control(without PFF). PFF had no effect on M-CSF mRNA expression in MLO-Y4 cells. These results suggest that osteocytes can regulate bone remodeling by communication with osteoclast precursors and osteoblasts and that osteocytes can communicate mechanical signals to other cells.