To identify whether higher expression of carboxylesterase (CbE) E4 in Myzus persicae is due to gene duplication, gene copy number was determined by quantitative real-time PCR. In addition, to determine the actual protein concentration of CbE E4 and it activity, Western blotting and activity staining were conducted. CbE gene copy number was highly correlated with carbamate resistance ratio (r2=0.934). However, CbE E4 expression level was little correlated with insecticide resistance ratio (r2<0.046) and no apparent correlation was observed among the gene copy number, protein quantity and total activity of CbE E4. Therefore, it was assumed that not only quantitative changing but also qualitative alteration of CbE E4 occurred in M. persicae. To investigate any potential alteration of CbE E4, mutation survey was conducted by sequencing of CbE E4 from various local strains of M. persicae. G137D and W251L mutations have been known as the main mutations associated with structural change leading to resistance. Interestingly, a new G134C mutation, which is in proximity of G137D mutation, was identified in the oxyanion hole of CbE E4. To predict the functional role of this mutation in resistance, 3-dimensional structure modeling was conducted. In summary, CbE E4 appears to be involved in resistance to both pyrethroids and carbamates as a nonspecific hydrolase or sequestration protein in M. persicae.

Zizyphi spinosi semen (Z. spinosi) has been used in traditional Chinese medicine for the treatment of rheumatoid arthritis and wounds. However, toxicity in high doses was often observed due to the presence of alkaloids. This study was conducted to investigate the potential genotoxicity of Z. spinosi in vitro and in vivo. This was examined by the Bacterial reverse mutation (Ames) test using Salmonella typhimurium TA98, TA100, TA1535, TA1537 and Escherichia coli WP2uvrA, Chromosomal aberration was investigated using Chinese hamster lung cells and the micronucleus test using mice. Z. Spinosi did not induce mutagenicity in the Ames test, and it did not produce chromosomal aberration in Chinese hamster lung cells with and without metabolic activation, nor in the micronucleated polychromatic erythrocytes in the bone marrow cells in mice. Based on these results, it is concluded that Z. spinosi does not have mutagenic potential under the conditions examined in each study.

The green peach aphid, Myzus persicae (Sulzer), is one of the most serious pest in cabbage cultivation. Field survey was carried out to know the insecticide resistance levels in five main cabbage cultivation regions (Pyeong-chang, Hong-cheon, Bong-wha, Mu-ju and Je-ju) in 2009~2011. The green peach aphid can resist a wide range of insecticides in five surveyed local populations. Among the nine tested insecticides, four chemicals (methomyl, bifenthrin, pymetrozine and flonicarmid) showed less than 60% mortality in the recommended concentration in most years of local populations. Multi resistant (MR) strain was selected from these populations and modified AChE (MACE: StoF mutation), MtoL mutation in para-type sodium channel, esterase over-expression were observed in almost of all populations including MR strain. Especially, StoF and MtoL mutations were highly correlated with resistance ratio and population based quantitative sequencing results. Therefore, these results suggested that molecular-biology based resistance monitoring can applied resistance management in M. persicae.

The cotton aphid, Aphis gossypii (Glover), is one of the most serious pest in seed potato and various vegetable cultivation. The imidacloprid-resistant strain (IR) was over 300-fold more resistant to imidacloprid compared to a susceptible strain (S) as judged by LC50 values. A highly imidacloprid-resistant local field population (L) was collected from cucumber at Gangwha island in 4th August 2011. Even though neonicotinoid insecticides especially imidacloprid were sprayed six times during June and July, aphid density was too high to be counted. To identify differentially expressed genes in IR or L, comparative transcriptome analyses based on GS-FLX were conducted using total RNAs extracted from IR, L and S strains. Futhermore, to search the resistance associated proteins in IR or L, comparative proteome analyses based on 2DE were conducted using total proteins extracted from IR, L and S strains. Few common candidate genes detected among IR and L such as ABC genes. Comparison of the nucleotide sequence of six nicotinic acetylcholine receptor (nAChR) subunit (alpha 1-5, beta 1) genes from IR, L and S strain revealed a point mutation in the loop D region of the nAChR beta 1 subunit of the IR, causing an arginine to threonine substitution (R81T). These mechanisms also reported in Myzus persicae and this amino acid change confers a vertebrate-like character to the insect nAChR and results in reduced sensitivity to neonicotinoids.

Malignant pleural effusion (MPE) and blood samples can be used as a practical source for detection of epidermal growth factor receptor (EGFR) mutations in patients with advanced non-small cell lung cancer. We compared EGFR mutation status of cell blocks, cell-free fluid of MPE, and plasma from patients with lung adenocarcinoma. We obtained paired samples of MPE and plasma from 14 pathologically-confirmed lung adenocarcinoma patients. Peptide nucleic acid (PNA)-mediated real-time polymerase chain reaction (RT-PCR) clamping was performed for determination of EGFR mutation status. EGFR mutations were detected in five (35.7%) cell blocks of MPE, which showed results identical to those of the corresponding cell-free fluid, whereas mutations were detected in the plasma of only two (40.0%) of the five patients. Of seven patients treated with EGFR tyrosine kinase inhibitors (TKIs), EGFR mutations were detected in cell blocks, cell-free fluid of MPE, and plasma for only one of the four patients who responded to EGFR TKIs, while mutations were detected only in cell blocks of MPE and cell-free fluid of the three remaining patients. Our results suggest that detection of EGFR mutations in cell-free pleural fluid from lung adenocarcinoma patients using highly sensitive methods may be feasible, but that analysis of free plasma may lead to undetected mutations and misdiagnosis.

Our aim was to investigate the genotoxicity of ambient air in the Krakow area after Fukushima Nuclear Power Plant (NPP) accident and compare with results from Chernobyl fallout. For the detection of ambient air genotoxicity the technique for scr

The two-spotted spider mite, Tetranychus urticae Koch (Acari: Tetranychidae), is one of the most important pest species devastating many horticultural, ornamental crops and fruit trees. Difficulty in managing this mite is largely attributed to its ability to develop resistance to many acaricides. Development of 3,700 folds resistance to etoxazole was found in the population of T. urticae collected from rose greenhouses in Buyeo, Chungnam Province in August 2000. This population has been selected for eleven years with etoxazole (over 500 times), and increased over 5,000,000 fold in resistance as compared with susceptible strain (S). Etoxazole-resistant strain was shown to be maternally inherited. The objective of this study was to determine whether resistance of T. urticae to etoxazole was linked with point mutations in the mitochondrial gene. DNA sequencing of cytochrome c oxidase subunit I (COX1), COX2, COX3, cytochrome b (CYTB), NADH dehydrogenase subunit 1 (ND1), ND2, ND3, ND4, ND5, and ND6 were analyzed by comparing two isogenic etoxazole-susceptible (EtoS) and etoxazole-resistant (EtoR) strains. As a result, all genes revealed no point mutations between the two strains.

The molecular mechanisms and genetics of abamectin resistance mediated by target site insensitivity in the two-spotted spider mite, Tetranychus urticae, were investigated by comparing two isogenic AbaS and AbaR strains. Cloning and sequencing of full-length cDNA fragments of GABA-gated chloride channel genes revealed no polymorphisms between the two strains. However, sequence comparison of the full-length cDNA fragment of a T. urticae glutamate-gated chloride channel gene (TuGluCl) identified a G323D point mutation as being tentatively related with abamectin resistance. In individual F2 progenies obtained by backcrossing, the G323D genotype was confirmed to correlate with abamectin resistance. Bioassays using progeny from reciprocal crossings revealed that the abamectin resistance trait due to TuGluCl insensitivity is incompletely recessive.

The stable fly, Stomoxys calcitrans L., is an important pest of livestock. Stable flies are considered as mechanical vectors of veterinary disease. Pyrethroids and organophosphates have been widely used for stable fly control. To establish resistance monitoring molecular tool, we isolated the partial cDNA and genomic fragments of voltage-sensitive sodium channel and acetylcholinesterase genes encompassing the well known conserved sites for resistance-associated mutations. To examine the current status of stable fly resistance to pyrethroids and organophosphates mediated by the nerve insensitivity mechanism in Korean population of S. calcitrans, DNA-based genotyping in conjunction with residual contact vial (RCV) bioassay were conducted with 11 representative regional field populations. No resistance-associated mutations were detected in these S. calcitrans populations, suggesting that these populations are likely still susceptible to both pyrethroids and organophosphates. Establishment of RCV bioassay protocol and availalbility of target site sequence information will greatly facilitate resistance monitoring of S. calcitrans in the field.

Molecular mechanisms of monocrotophos resistance in the two-spotted spider mite (TSSM), Tetranychus urticae Koch, were investigated. The resistant (AD) strain showed ca. 3,565-fold resistance compared to a susceptible (UD) strain. No significant differences in the esterase and gluthathion-S-transferase activities were found between two strains whereas AD showed a 1.9-fold higher mixed function oxidase activity. Acetylcholinesterase (AChE) inhibition assay revealed that the AChE from AD strain is 91-fold less sensitive to monocrotophos, suggestive of the target site insensitivity mechanism. Three point mutations (G228S, A391T and F439W/Y) in the AChE gene (tssmace) appeared to primarily contribute to the reduced sensitivity of AChE as judged by the correlation study of mutation frequency versus resistance levels (LC50) of several field populations. The resulting correlation coefficients of the G228S and F439W mutations were 0.711 and 0.300, respectively, suggesting that G228S mutation may play a more significant role in resistance. The A391T mutation, saturated in all field populations examined, appears to provide a base line resistance.

본 연구는 감마선 조사 및 배양방법을 이용하여 유 용한 변이체를 선발하고, 그 특성을 관찰하기 위해 수 행되었다. 기내 신초수의 증식은 MS 배지 내에 NAA 0.2mg·L-1에 BA의 농도가 1.0mg·L-1로 증가할수록 양호한 반면 신초의 길이와 뿌리형성율은 감소되었다. 계대배양을 3회 수행한 후 온실에서 순화 및 삽목하였다. 감마선으로 조사된 총 370 개체들은 감마선의 조 사선량에 관계없이 토양이식 후 생존율이 97% 이상이 었다. 조사된 개체들 중 변이의 빈도는 감마선의 선량 이 증가할수록 높아졌다. 50Gy 조사구에서는 화색과 화형의 변이가 각각 28.2와 15.4%로 확인되었다. 화 색과 화형은 다양한 변화를 나타냈으며 줄기색 또한 변화되었다. 예를 들면 흰색에 옅은 자주색의 관상화는 흰색, 적자색, 노랑색 또는 연분홍색 등으로, 흰색이었 던 설상화는 연한 자주색이나 적자색으로 바뀌었다. 설 상화의 길이와 폭 및 화경의 크기가 달라진 개체 및 줄기에서 안토시아닌 색소가 제거된 개체도 관찰되었 다. 본 연구결과 국화 ‘Argus’에서 기내 배양체에 30- 50Gy의 감마선 조사에 의해 화색, 화형 및 줄기색의 다양한 돌연변이체를 선발할 수 있었다.

Cherubi sm is a ra re autosoma l dominant inherited condi tion caused by mutations in the c-Abl-binding protein SH3BP2. 1t is characteri zecl by mul t iple cystic giant cell lesions of the jaw appearing in early childhood with stabi li zation and rcmi ssion after pubcr ty, In thc present study, genomic DNA was purified f rom a blood sample obtained from the patient a ncl pa rents a ncl used f'or di rect sequence analysis of the SH3BP2 gene, 1n addit ion, a sample of the lesion was used f0 1" hi stologic ancl immunoh is toche mical purposes, Histology revealed a proliferation of spindle s haped fibroblas t ic cells and irregu la ry dis persed multinucleated giant cell s , The multinucleated giant cells proved posit ive for CD68 and TRAP, Ge nomic DNA sequencing f'ou ncl a missense mutation Pro418Arg in exon 9 of the SH3BP2 gene of the patient and the mother, Theref'ore, the mul t inucleated giant cells are basically osteoclastic in nature, Additionally, as the PI'o418Arg mutation had been repol' ted as caus ing cherubism, it represents a mutational hotspot,

Hereditary dentin defects consist of dentin dysplasia(DD) and dentinogenesis imperfecta(DI). The DI associated with osteogenesis imperfecta has been classified as DI type I, whereas isolated inherited defects have been categorized as DI types II and III. However, whether DI type III should be considered a distinct phenotype or a variation of DI type II is debatable. Recent genetic findings have focused attention on the role of the dentin sialophosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin. We have identified a novel mutation(c.727G → A, p.D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III. The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth. These findings suggest that DI type II and III are not separate diseases but rather the phenotypic variation of a single disease.

The efficiency to detect mutagenicity of the system using a specific locus mutation of Bombyx mori was examined and improved. In the system, mutagenicity could be detected by the egg colour manifested by the pe and/or re genes, which is a kind of recessive visible mutation of the insect. Among tested four mutagens, MMC had specially high sensitivity in the oocytes of silkworm and EMS had in the spermatozoa. PCB and dioxin showed a positive effect in both the oocytes and spermatozoa. In a consequence of sensitivity of mutagen by mating number of male moth of B. mori, treated mutagen, there was no difference between one mating - and three mating - male moth in sensitivity of mutagen. Sun3ho, B. mori variety, which showed high sensitivity to mutagens was improved in the major characteristics by crossing of C5 and N12.

Hereditary dentin defects consists of dentin dysplasia(DD) and denti nogenesis imperfecta(Dr) ‘ The Dl associated with osteogenesis imperfecta has been classified as DI type 1. whereas isolated inherited defects have been categori zed as DI types II and III , However‘ whether DI type III should be considered a distinct phenotype 01' a variation of DI type 1I is debatable , Recent genetic findings have focused attention on the role of the dentin sialo phosphoprotein(DSPP) gene in the etiology of inherited defects of tooth dentin, We have identified novel mlltation( c,727G - > A, p,D243N) at the 243th codon of exon 4 of the DSPP gene in a Korean patient with DI type III The radiographic and histologic features of the patient revealed the classic phenotype of shell teeth These findings sllggest that DI type II and III are not separate diseases bllt rather the phenotypic variation 01' a s ingle disease

Odontogenic ghost cell tumor (OGCT) is considered as a neoplastic counterpart of the calcifying odontogenic cyst (COC). β-catenin mutations have been described in COC suggesting a critical role in its histogenesis. In this study, we report a patient with OGCT contains a missense mutation on codon 3 (ACT -> TCT). Immunohistochemistry showed nuclear, cytoplasmic, and membranous accumulation of β-catenin in the tumor cells. TUNEL assay showed positive signals in nucleated cells adjacent to the ghost cells. Our data suggest that β-catenin plays an important role in the tumorigenesis of OGCT. OGCT may developed by improper differentiation process coordinated by WNT signaling pathway. Further studies are needed to determine a genotypic/phenotypic characteristics of ghost cell containing odontogenic lesions.

산삼배양추출물의 세균에서의 돌연변이 유발성 검색을 위하여 Salmonella typhimurium의 히스티딘 요구성 균주 TA100, TA1535, TA98 및 TA1537의 4개의 균주와 대장균 Eschericha coli의 트립토판 요구성 균주인 WP2 uvrA를 이용해 복귀돌연변이 시험을 실시하였다. 시험물질은 멸균생리식염수에 용해하여 처리하였다. 대사활성계 적용 및 미적용시 모든 균주에 대해 0, 62, 185, 556, 1,667 및 5,000 μg/plate의 범위를 설정하고 각각 음성 및 양성대조군으로 시험군을 구성해 본 시험을 실시하였다. 시험 결과 모든 균주에서 최고농도에 이르기까지 집락수의 일관성 있는 증가는 나타나지 않았다. 이상의 결과를 종합할 때, 시험물질 산삼배양추출물은 본 시험조건 하에 사용한 시험균주들의 복귀돌연변이를 유발하지 않는 것으로 사료된다.

본 연구는 국제정서사진체계(IAPS) 사진 중 이차원 감성을 잘 표현한다고 판단되는 9장의 사진을 선정하고, 이 사진 자극으로 유발된 감성의 변화를 심리ㆍ생리적으로 측정한 후 이차원 감성의 한 축인 각성/이완의 감성변화를 심리ㆍ생리적으로 변별 가능한지를 검증하는 것을 목표로 한다. 각성/이완의 감성변화를 대변하는 Eelectrocardiogram (ECG), Galvanic Skin Response (GSR), Skin Temperature, Respiration 등의 자율신경계 반응을 측정하였다. 자극 제시 후에 주관적 평가를 실시한 바, 각성 〉 불쾌/각성 및 쾌/각성 〉 중립 〉 이완 순서의 4단계로 각성도 감성을 구분할 수 있었다 자율신경계 반응은 각성, 불쾌/각성, 쾌/각성 〉 중립 〉 이완 순서의 3단계로 구분이 가능하였다. 본 연구는 한국인의 이차원 감성을 표현할 수 있는 9장의 사진을 IAPS로부터 선정하였으며, 이차원 감성의 한 축인 각성/이완의 감성 변화를 심리ㆍ생리적평가로부터 변별할 수 있다는 사실을 확인하였다.