Microfluidics based on nanobio sensors technologies can provide convenient and accurate diagnosis tools. In this talk, we present recent developments of nanobio sensors & diagnosis chip using microfluidics, with special emphasis on disposable plastic devices format. In detail, we overview of the common methods used in the fabrication of polymer microfluidic systems, including replica and injection mold-ing. Also we explain the different methods by which on-chip operations—such as the pumping and valving of fluid flow, the mixing of different reagents, and the separation and detection of different biochemical species implemented in a microfluidic format. Finally, a few select biotechnological applications of microfluidics are presented to illustrate both the utility of this technology and its potential applications with insect models in the near future.

Sirtuin proteins are evolutionary conserved Sir2-related NAD+-dependent deacetylases and regulate many of cellular processes such as metabolism, inflammation, transcription, and aging. Sirtuin contains activity of either ADP-ribosyl-transferase or deacetyltranfease and their activity is dependent on the localization in cells. However, the expression pattern of Sirtuins has not been well studied. To examine the expression levels of Sirtuins, RT-PCR was performed using total RNAs from various tissues including liver, small intestine, heart, brain, kidney, lung, spleen, stomach, uterus, ovary, and testis. Sirtuins were highly expressed in most of tissues including the testis. Immunostaining assay showed that Sirt1 and Sirt6 were mainly located in the nucleus of germ cells, spermatocytes, and spermatids in the seminiferous tubules, whereas Sirt2 and Sirt5 were exclusively present in the cytoplasm of germ cells and sperma-tocytes. Our results indicate that Sirtuins may function as regulators of spermatogenesis and their activities might be dependent on their location in the seminiferous tubules.

This studies was conducted to define community structure and diversity of epigeal spiders in apple orchards under different farming practices (organic and conventional). In 11 commercial apple orchards, epigeal spider community structure was investigated during 2012-2014 (3 orchards, 2012; 3 orchards, 2013, 5 orchards, 2014). Epigeal spiders were collected by pitfall traps from April to October, 2012 in Danyang-gun, Chungcheongbuk-do; 2013 in Muju-gun, Jeollabuk-do; 2014 in Cheongsong-gun and Uiseong-gun, Gyeongsangbuk-do, Korea. Sampled spiders were identified to the species level. Biodiversity was analyzed with species richness (total numbers of spider species), abundance (total numbers of individuals) and Shannon-Wiener diversity index (H΄) for each site and compared between organic and conventional orchards. In all regions, abundance of the spider communities were relatively higher in organic orchards than in conventional orchards. But, no significant difference in species richness and specie diversity (H΄) between organic and conventional orchards. Lycosidae was the most dominant family in all orchards regardless farming practices. However, Linyphiidae dominated in organic orchards only. The most dominant species were Pardosa laura (Lycosidae), Erigone koshiensis (Linyphiidae) in organic, Piratula procurvus (Lycosidae) in conventional orchards.

Epigeic spiders (Araneae) were surveyed in apple orchards under different farming practices (organic and conventional) in Muju-gun, Jeollabuk-do, Korea. The study was conducted from April to October in 2013 with 2 weeks interval using pitfall traps. Collected spiders were identified as 20 species of 6 families from 819 individuals in organic orchardⅠ, 19 species of 10 families from 759 individuals in orchardⅡ and 16 species of 8 families from 209 individuals in conventional orchard for from 1,787 captured spiders. Biodiversity was analyzed with species richness, abundance and Shannon diversity index (H΄). Species richness was statistically higher in organic orchard than in conventional orchard. Organic orchardⅡ comprised the greatest abundance followed by organic orchard and conventional. But species diversity was the highest in organic orchardⅡ, intermediate in conventional orchard and the lowest in orchardⅠ. Lycosidae was the most dominant family in all orchards regardless farming practices. However, Linyphiidae dominated in organic orchards only. The most dominant species were Erigone koshiensis (Linyphiidae) in organicⅠ, Pardosa laura (Lycosidae) in organicⅡ and Arctosa pungcheunensis (Lycosidae) in conventional orchard.

In apple orchards under different farming practices (organic and conventional), arthropod community structure was investigated. Arthropods were collected with pitfall traps, band traps, yellow sticky traps and pheromone traps from May to December, 2012 in Danyang-gun, Chungcheongbuk-do, Korea. Biodiversity was analyzed with species richness and Shannon diversity index (H΄). Sampled arthropods were identified 37 families in 14 orders from 21,618 individuals in organic orchard-1 and 33 families in 10 orders from 11,318 individuals in organic orchard-2 and 35 families in 13 orders from 27,711 individuals in conventional orchard. Species richness was relatively highest in organic orchard-1, intermediate in conventional orchard and lowest in organic orchard-2. In ecologically functional group, abundance of pests was higher in conventional orchard than in organic orchards due to the high occurrence of Phyllonorycte ringoniella (Gracillariidae) and Lyonetia prunifoliella (Lyonetiidae). Organic orchards showed higher abundance, species richness and species diversity of epigeic predators and those of parasitoids above ground than conventional orchard. Abundance of detritivores, mainly collembola, were higher in organic orchard than in conventional orchard.

Mamestra brassicae (cabbage moth) is a common European moth of the order Lepidoptera and the family Noctuidae. The larval stage is highly polyphagous and is known to feed on more than 70 species of host plants from 22 families, including Brassica species, lettuce, onion, potato, pea, tomato and apple. M. brassicae has become a significant pest also in Asia due to the damage caused to agriculturally and economically important Brassica crops. It is difficult to control M. brassicae using chemical insecticide because of its rapid development of resistance. The objective of our study, therefore, was the mass production and formulation of a local strain of M. brassicae nucleopolyhedrovirus-K1 (MabrNPV-K1) for the development of viral insecticide to control it. In production efficiency of MabrNPV-K1 using M. brassicae larvae, the mortality of the 3rd instar larvae was 100% when inoculated with 1.0 × 105 PIBs/larva and the yield of MabrNPV-K1 was maximal. Regarding the mortality, yield of polyhedra, inoculation doses and required time, the 1.0 × 104 PIBs/larva at 30°C was determined as optimal conditions producing polyhedra efficiently. To formulate MabrNPV-K1, feeding toxicities of various supplements including spreader and ultraviolet (UV) -protectant were determined. Tinopal UNPA-GX which is UV-protectants was effective for protection of polyhedra from UV and showed the increased mortality when added with 1% concentration. Other supplements did not influence significantly the mortality of MabrNPV-K1. Formulated MabrNPV-K1 with several supplements showed higher pathogencity than un-formulated MabrNPV-K1.

The diamondback moth, Plutella xylostella is a one of the most important pests of various cruciferous crops and has a geographically wide ranging habitat. The heavy dependence on chemical pesticides has created severe pesticide resistance problems. In recent years, Bacillus thuringiensis product have been widely used for P. xylostella control bus genetic resistance in populations to some B. thuringiensis strains, compounded by cross-resistance to several different B. thuringiensis toxins, has also been identified. Such recent resistance problems serve to emphasize the urgent need for alternative control agents and their use within an integrated pest management approach. Baculoviruses have been used as agents for the biological control of certain insect pest species. the granuloviruses (GVs), based on the structure of the occluded virus and the occlusion body (OB). Several reports have showed P. xylostella granulovirus (PxGV) as a promise control agent for P. xylostella. However, it is very difficult to study GV because its OB, granule, has very small size and could be observed exactly under the electron microscopy (EM). This study was performed to develop rapid quantification method for granule of PxGV. After the exact quantification of granule with latex beads using EM, the universal extraction method of viral DNA was established for consistent experiment. The number of granules was calculated by the quantification of PCR products for granuline gene using spectrophotometer and densitometer. This novel calculation method for granule would be useful to study GV.

Three acetylcholinesterases (AChEs) were identified from the pinewood nematode, Bursaphelenchus xylophilus. Sequence comparison with known AChEs in conjunction with three-dimensional structure analysis suggested that all BxAChEs share typical characteristics of AChE at the major catalytic structures. BgAChE3 was most predominantly transcribed and then followed by AChE1 and AChE2. Immunohistochemistry using anti-BxAChEs antibodies revealed that BxAChE1 is most widely distributed whereas BxAChE2 exhibits more localized distribution in neuronal tissues. BxAChE3 was detected from entire body together with some limited tissues, including mouth parts and alimentary lining, and determined to be the only soluble AChE, suggesting its localization in hemolymph or/and extracellular space. Kinetic analysis of in vitro expressed BxAChEs revealed that BxAChE1 has the highest substrate specificity whereas BxAChE2 has the highest catalytic efficiency with BxAChE3 having the lowest catalytic efficiency. Interestingly, presence of BxAChE3 in the pool of BxAChEs significantly reduced the inhibition of BxAChE1 and BxAChE2 by inhibitors. Knockout of BxAChE3 by RNAi significantly increased the toxicity of nematicides, suggesting the protective role of BxAChE3 against these toxicants. Based on several features, including tissue distribution, expression level, substrate kinetics and inhibition property, it appeared that BxAChE1 is the major AChE with the function of postsynaptic transmission whereas BxAChE3 has been evolved to acquire the function of chemical defense, perhaps intrinsically against secondary toxic compounds from host pine trees, such as α-pinene and limonene. BxAChE2 appears to play a role in post-synaptic transmission in specialized neurons but its detailed physiological function still remains to be elucidated.

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

Somatic cell nuclear transfer in cattle has limited efficiency in terms of production of live offspring due to high incidence of fetal failure after embryo transfer to recipients. Such low efficiency of cloning could possibly arise from abnormal and poorly developed placenta. In the present study the placental proteome in late pregnancy established from in vitro fertilization (IVF) and nuclear transfer (NT) was analysed. Proteome alternation was tested using two-dimensional polyacrylamide gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI- TOF). Comparing placenta from NT embryos to those from IVF counterparts, significant changes in expression level were found in 18 proteins. Of these proteins 12 were not expressed in NT placenta but expressed in IVF counterpart, whereas the expression of the other 6 proteins was limited only in NT placenta. Among these proteins, cytokeratin 8 and vimentin are considered to be involved in regulation of post-implantation development. In particular, cytokeratin 8 and vimentin may be used as makers for placental development during pregnancy because their expression levels changed considerably in NT placental tissue compared with its IVF counterpart. Data from 2-DE suggest that protein expression was disorientated in late pregnancy from NT, but this distortion was eliminated with progression of pregnancy. These findings demonstrate abnormal placental development during late pregnancy from NT and suggest that alterations of specific placental protein expression may be involved in abnormal function of placenta.