The resistance levels against carbamates (CB) and organophosphates (OP) were determined through bioassay and quantitative sequencing (QS) methods in 16 field populations of Nilaparvata lugens. The resistance levels to CB and OP by bioassay were 1.3~47.5-fold and 1.4~14.4-fold higher than a susceptible strain, respectively. The QS protocol was established to determine the allele frequencies of eight point mutations on acetylcholinesterase putatively associated CB and OP resistance. The allele frequencies of four mutations in local populations (G119A, F/S330Y, F331H and I332L) ranged from ca. 0.0~51.7%, 1.0~44.3%, 8.5~57.3% and 7.12~56.6%, respectively. The average prediction limits were –9.6±5.1~7.7±2.9%. The F330Y, F331H and I332L were tightly linked each other, suggesting these mutations may occur simultaneously. In the correlation analysis, G119A was not well correlated with both insecticides (r2= less 0.25), whereas F/S330Y, F331H and I332L showed better correlation with the resistance levels of carbamate (r2=0.590) than organophosphate (r2=0.235). This finding indicates that F/S330Y, F331H and I332L mutation frequencies are suitable for detecting carbamate resistance in N. lugens. QS will be applicable for the rapid monitoring of resistance levels to CB insecticides in N. lugens.

A carbofuran-resistant strain (CAS) showed ca. 41.1- and 15.1-fold resistance compared to a susceptible strain (SUS) and a non-selected field strain (FM), respectively. Enhanced activities of carboxylesterase and P450 were found as ca. 3- and 1.6-fold higher in CAS strain, suggesting these enzymes play a minor role in carbofuran resistance. Interestingly, the insensitivity of acetylcholinesterase (AChE) to carbofuran was revealed to be ca. 5.5- and 3.7-fold higher in CAS strain compared to, indicating that AChE insensitivity mechanism is associated with carbofuran resistance. In the western blot analysis, two kinds of AChEs were found and type-1 AChE (Nlace1) was identified as the major AChE in N. lugens. The open reading frame of Nlace1 is composed of 2,106 bp (ca. 78 Kd) and revealed 52.5% and 24.3% identity compared with Nephotettix cincticeps and Drosophila melanogaster, respectively. In the screening of point mutations, four amino acid substitutions (G119A, F/S330Y, F331I and H332L) were identified in the CAS strain that likely contribute to the AChE insensitivity. The allele frequencies of these mutations increased in the survived populations following the selection by LC50 of carbofuran, confirming that they are in fact associated with reduced sensitivity to carbofuran in N. lugens. These point mutation can be useful for the monitoring of resistance levels in conjunction with QS methods.

We identified and characterized the full-length cDNA sequences encoding two acetylcholinesterases (ClAChE1 and ClAChE2) and a salivary gland-specific cholinesterase (ClSChE) from the common bed bug, Cimex lectularius. All three cholinesterase genes (Clac1, Clace2 and Clsce) have conserved motifs, including a catalytic triad, a choline binding site and an acyl pocket. Phylogenetic analysis showed that ClAChE1 belongs to the insect AChE1 clade, whereas ClAChE2 belongs to the insect AChE2 clade. ClSChE was grouped into the clade containing all AChE1s, suggesting its paralogous relationship to ClAChE1. Transcription levels of Clace1 were higher than those of Clace2 in all tissues examined, including the central nervous system (CNS). In contrast, the Clsce transcript was not detected in the CNS but specifically found in the salivary gland in much higher levels (>3000 fold) than those of Clace1 and Clace2. Western blot analysis using anti-ClAChE antibodies in conjunction with activity staining revealed that ClAChE1 is more active than ClAChE2 whereas ClSChE has little enzyme activity. Three-dimensional structure modeling suggested that ClAChEs and ClSChE shared structural similarities, but had some differences in the residues forming the acyl pocket and oxyanion hole. The current findings should provide valuable insights into the evolution and functional diversification of insect cholinesterase.

Capsaicin β-glucoside was isolated from the feces of Helicoverpa armigera, H. assulta and H. zea that fed on capsaicin-supplemented artificial diet. The chemical structure was identified by NMR spectroscopic analysis as well as by enzymatic hydrolysis. The excretion rates of the glucoside were different among the three species; those in the two generalists, H. armigera and H. zea, were higher than in a specialist, H. assulta. UDP-glycosyltransferases (UGT) enzyme activities measured from the whole larval homogenate of the three species with capsaicin and UDP-glucose as substrates were also higher in the two generalists. Compared among five different larval tissues (labial glands, testes from male larvae, midgut, the Malpighian tubules, and fatbody) from the three species, the formation of the capsaicin glucoside by one or more UGT is high in the fat body of all the three species as expected, as well as in H. assulta Malpighian tubules. Optimization of the enzyme assay method is also described in detail. Although the lower excretion rate of the unaltered capsaicin in H. assulta indicates higher metabolic capacity toward capsaicin than in the other two generalists, the glucosylation per se seems to be insufficient to explain the decrease of capsaicin in the specialist, suggesting H. assulta might have another important mechanism to deal with capsaicin more specifically.

Autophagy is a process of intracellular bulk protein degradation, in which the accumulated proteins and cytoplasmic organelles are degraded. It plays important roles in cellular homeostasis, apoptosis, and development, but its role during early embryo development remains contentious. Therefore, in the present study, we investigated the effects of 3-methyladenine (3-MA) on early embryonic development in pigs. we also investigated several indicators of developmental potential, including mitochondrial distribution, genes expressions (autophagy-, apoptosis- related genes), apoptosis and ER-stress, which are affected by 3-MA. After in vitro maturation and fertilization, presumptive pig embryos were cultured in PZM-3 medium supplemented with 3-MA for 2 days at 39℃, 5% CO2 in air. Developmental competence to the blastocyst stage in the presence of 3-MA was gradually decreased according to increasing concentration. Thus, all further experiments were performed using 2 mM 3-MA. Blastocysts that developed in the 3-MA treated group decreased LC3-II intensity and expressions of autophagy related genes than those of the untreated control, resulting in down-regulates the autophagy. Terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling (TUNEL) showed that the number of containing fragmented DNA at the blastocyst stage increased in the 3-MA treated group compared with control (6.0±1.0 vs 3.3±0.6, p<0.05). Also, the expression of the pro-apoptotic gene Bax increased in 3-MA treated group, whereas expression of the anti-apoptotic gene Bcl-XL decreased. Mito Tracker Green FM staining showed that blastocysts derived from the 3-MA treated group had lower mitochondrial integrity than that of the untreated control, resulting in decrease the embryonic qualities of preimplantation porcine blastocysts. Then, the expression of the spliced form of pXBP-1 product (pXBP-1s) increased in 3-MA treated group, resulting increase of ERstress. Taken together, these results indicate that inhibition of autophagy by 3-MA is closely associated with apoptosis and ER-stress during preimplantation periods of porcine embryos.

본 논문에서는 산업 현장에서 비전기술을 이용한 객체의 검사나 분류에 사용되는 빠르고 효율적인 최소 사각형 근사화 방법을 대해서 소개한다. 주어진 객체의 최소 사각형 근사를 위해서 객체의 전체 외곽선을 사용하지 않고 n 개로 샘플링된 에지 점들에 대해서만 각도 히스토그램을 생성하고 이를 이용하여 객체의 장축과 단축을 계산한다. 최종 근사 사각형을 찾기 위해 이전 단계에서 얻어진 장축과 단축을 중심으로 하는 4개의 모서리점을 갖는 사각형을 찾는다. 이후에, 잡음에 의한 오차를 줄이기 위해 첫 번째 사각형의 각도를-1∼+1로 변형하여 추가적으로 2개의 후보 사각형을 생성한 후, 3개의 추정된 사각형과 원본 영상의 외곽선의 매칭 정도를 측정하여 오류가 최소화 되는 사각형을 최종 사각형 영역으로 선택한다. 제안된 방법을 몇 개의 실험데이터에 대해 실험한 결과 기존의 방법에 비해 빠르고 효율적인 최소 사각형 근사가 가능하였다.

A 20' X 20' region around 30 Doradus in the Large Magellanic Cloud (LMC) is observed and analyzed in the near-infrared. We obtain polarimetry data in the J, H, and Ks bands using the SIRIUS polarimeter SIRPOL at the Infrared Survey Facility 1.4 m telescope. We measure the Stokes parameters of 2562 point-like sources to derive the degree of polarization and the polarization position angles. We discuss the statistics of the groups classified by color-magnitude diagram and proper motions of the sources, in order to separate the Galactic foreground sources from those present in the LMC. We notice that groups classified by the proper motion data show a tendency towards different polarimetric properties.

The forest of Korea had been severely degraded since early 1900s until 1950s. Korean Government has successfully accomplished the reforestation works since 1960s. However, some plantations showed poor survival and growth caused by ignoring site characteristics in selecting plantation species and lack of tending works such as thinning. The natural regeneration of indigenous species, such as Quercus species and Pinus densiflora Siebold & Zucc., were examined in the plantations of Pinus koraiensis Siebold & Zucc. and Larix kaempferi Fortune ex Gordon. Quercus species regenerated mainly by sprouting while P. densiflora regenerated naturally from a few mother trees that remained in the plantations. P. koraiensis showed poor survival (IVI ≤ 25%) and suppressed growth (height ≤ 3 m and DBH ≤ 3 cm at 20 year-old) by Quercus species or P. densiflora in the plantation areas, however had high survival (IVI ≥70%) and growth (8 m height and 14.1 cm DBH at 20 year-old) in areas where silvicultural practices were conducted. L. kaempferi showed good survival (IVI ≥40%) and growth (17.2 m height and 16.3 cm DBH at 30 year-old) mostly in valley areas, while it was nearly dead (IVI ≤ 10%) in ridge or ridge-slope areas and was replaced by indigenous species such as Quercus species (IVI ≥ 25～55%) or P. densiflora (IVI ≥ 18～50%).

목적: International Association of Contact Lens Educators(IACLE)가 실시하는 STE결과를 분석하여 한국에서의 콘택트렌즈 교육 실태를 알아보고자 한다. 방법: 2010년 IACLE에서 주관한 STE에 참여한 12개 대학 안경광학과 학생들의 시험성적을 분석하여 대학 및 국가간 비교를 실시하였다. 결과: 12개 대학의 평균은 100점 만점을 기준으로 하여 54.01점이었으며 평균 합격률은 62.75%였다. 문항을 주제별로 7개 부류로 나누어 분석한 결과 평균 점수는 각각 전안부 해부 및 생리 48.73점, 콘택트렌즈 디자인 및 관리 46.14점, 콘택트렌즈 피팅원리 49.08점, 환자 검사 및 콘택트렌즈 처방 73.37점, 콘택트렌즈 관리 및 유지 66.12점, 콘택트렌즈에 대한 안구 반응 55.45점, 콘택트렌즈 착용으로 인한 부작용 52.54점으로 나타났다. 교육기간에 따라 2,3년제 대학과 4년제 대학을 비교해 본 결과 2,3년제 대학의 평균 점수가 53.35점, 4년제 대학의 점수가 54.94점으로 큰 차이를 보이지 않았다. 국가간 비교에서는 중국, 인도, 프랑스, 독일에 비해 한국 학생들의 점수가 다소 낮은 것으로 확인 되었다. 결론: 평균점수와 합격률에 있어 학교 및 과목 사이에 편차가 확인 되었다.

Ovulation synchronization (ovsynch) has proved to increase the number of insemination in cattle by overcoming the problems of heat detection. The aim of this study was to do ovsynch in water buffaloes where heat detection is a major reproductive problem and to determine the conception rates after timed artificial insemination (TAI). Twenty cyclic buffaloes at 60 days postpartum were selected by examining 24 unobserved estrus buffaloes based on milk progesterone assay (progesterone concentration 1.0 ng/ml) from the Mymensingh district of Bangladesh. Ovsynch treatment regimen was started irrespective of the stage of estrous cycle. Gonadorelin (500 ) was injected intramuscularly at Day 0 followed by Alfaprostol (8 mg) at Day 7. A second injection of Gonadorelin was given at Day 9 and TAI was done with frozen semen from Mediterranean buffalo bulls at 16~20 hours of the second Gonadorelin injection. Milk progesterone ELISA at Day 10~12 post AI confirmed ovulation in 16 out of 20 (80%) buffaloes (progesterone concentration 1.0 ng/ml). High progesterone concentration ( 1.0 ng/ml) at Day 10~12 and Day 22~24 of AI showed pregnancy in six out of 20 (30%) buffaloes. Pregnancy was further confirmed by ultrasonography at Day 40 in these six buffaloes. In conclusion, ovsynch followed by TAI could be applied in cyclic buffaloes for overcoming the estrus detection problems; however, more studies are needed to increase the conception rate.

Edible mushrooms have been used and researched as medicinal ingredients. They improve immunity and contain excellent anticancer components with large amounts of minerals such as iron, calcium, and potassium. Due to this reason, it has been hailed as a raw material for functional foods. Especially, β-glucan, known to be contained in Ganoderma incidum Karst, Coriolus versicolor, and Phellinus linteus, was reported to inhibit proliferation of cancer cells by activating immune function (macrophages, natural killer cells and T-cells). In addition, mushroom polysaccharides dissolve in water but are undigested ingredients, resulting in stimulating the immune system as well as staying of parasympathetic nervous system in the stomach for a long time. As a result, they induce increase of T-cells and NK cell that attacks cancer cell and has effect on the discharge of body's waste products, blood purification, and constipation improvement. In this study, we report the culture characteristics of Sparassis crispa as to the medium growth compositions, yeast, and elicitor treatments to investigate the optimal condition for the highest β-glucan production in mushrooms.

Lycorma delicatula (White 1845), which has been recently introduced into Korea, is a notorious pest on grapes. This invasive insect has rapidly spread throughout central and southern Korea. To date, we have no behavioral or population genetics information, such as invasion routes and subsequent dispersal rates in Korea, to help understand and control populations of L. delicatula. Here, we have developed 15 novel microsatellite loci for L. delicatula. The isolated loci were polymorphic, with 2 to 19 alleles in 42 individuals from a single population in Cheonan. The analyses revealed that all 42 individuals had different multilocus genotypes with heterozygosity ranging from 0.214 to 0.866. Eleven of the 15 loci did not deviate significantly from Hardy-Weinberg equilibrium. The isolated markers will facilitate population genetic studies of L. delicatula.

Western blot analysis using acetylcholinesterase (AChE)-specific antibody was conducted to determine whether AChE gene (Tuace) duplication actually results in overproduction of AChE in Tetranychus urticae (TuAChE). The protein quantities of TuAChE in seven field-collected mite populations were precisely correlated with the copy numbers. To investigate the effects of each mutation on AChE insensitivity and possible fitness cost, eight variants of TuAChE were in vitro expressed using the baculovirus expression system. Kinetic analysis revealed that the Ala391Thr mutation did not change kinetic properties of AChE, whereas the Gly228Ser and Phe439Trp mutations significantly increased the insensitivity to monocrotophos. Moreover, when the Gly228Ser and Phe439Trp mutations are present together, insensitivity increased over a thousand-fold, showing that both mutations confer resistance in a synergistic manner. Presence of the mutations, however, reduced catalytic efficiency of AChE considerably, suggesting an apparent fitness cost in monocrotophosresistant mites. Reconstitution of the multiple copies of AChE having different compositions of mutations revealed that the catalytic efficiencies of the six-copy and two-copy AChEs (resembling the AD and PyriF strains of mite, respectively) were still lower but comparable to that of wildtype AChE. These finding clearly suggested that multiple rounds of Tuace duplication was needed to compensate the reduced catalytic activity of AChE caused by mutations.

A polydnavirus, Cotesia plutellae bracovirus (CpBV), is a symbiotic provirus to an endoparasitoid wasp, C. plutellae. When the wasp parasitizes its natural host, Plutella xylostella, larvae, CpBV viral particles are translocated to hemocoel of P. xylostella along with the wasp eggs. CpBV-ELP1 is encoded in a viral segment and expressed in the parasitized larvae during entire parasitization period. A recombinant baculovirus expressing CpBV-ELP1 was constructed and applied to a non-natural host, Spodoptera exigua, larvae. When the recombinant baculovirus was injected to hemocoel, CpBV-ELP1 was expressed in hemocytes as early as 2h postinjection and then later expressed in other tissues. When it was applied to diet, CpBV-ELP1 was expressed in midgut epithelium at 12 h and subsequently expressed in internal tissues. Both application methods of the recombinant baculovirus caused significantly higher mortality of S. exiguathan non-recombinant baculovirus. Interestingly, midgut epithelial cells expressing CpBV-ELP1 by infection of the recombinant baculovirus showed poor cell-cell interactions. Integrin, a cell surface molecule associated with cell-cell interaction, was cloned in S. exigua and was confirmed in its expression in the midgut epithelium. A hypothesis was raised that CpBV-ELP1 interrupts integrin function by direct binding or by blocking internal integrin signaling.