The present investigation establishes deltamethrin resistance in the common bed bug, Cimex lectularius, populationcollected from New York City (NY-BB). The mortality resistance ratio indicated that NY-BB population was 264-fold more resistant to 1% deltamethrin in contact bioassay compared to one insecticide- susceptible population collected in Florida (FL-BB). Specific enzyme activities (general esterase, glutathione S-transferase, and 7-ethoxycoumarin O-deethylase) of NY-BB were not statistically different from those of FL-BB, indicating that the metabolic factors are not associated with the deltamethrin resistance in NY-BB. Complementary DNA fragments that encoded the open reading frame of voltage-sensitive sodium channel α-subunit genes from the FL-BB and NY-BB were obtained by homology probing PCR and sequenced. Sequence alignment of the internal and 5’ and 3’ RACE fragments generated a 6500 bp cDNA sequence contig, which was composed of a 6084 bp open reading frame (ORF) encoding 2027 amino acid residues and 186 bp 5’ and 230 bp 3’ untranslated regions (5’ and 3’UTRs, respectively). Sequence comparisons of the complete open reading frames of the sodium channel genes identified two point mutations (V419L and L925I) that were presented only in the NY-BB population. L925I, located the intracellular loop between IIS4 and IIS5, has been previously found in the pyrethroid-resistant populations of whitefly (Bemisia tabaci) that was more than 100-fold resistant to fenpropathrin. V419L, located in the IS6 transmembrane segment, is a novel mutation. This evidence suggests that the two mutations are likely the major resistance-causing mutations in the deltamethrin-resistant NY-BB via a knockdown-type nerve insensitivity mechanism.

Innate immunity responses are triggered by the immune challenge and therefore involve signaling processes. The cellular response is initiated by hemocytes and mainly involves phagocytosis and encapsulation of intruders by these cells. To address whether Hc-STAT is activated upon bacterial challenge, we examined the subcellular location of STAT protein in hemocyte by immunostaining. A new insect member of the STAT family of transcription factors (Hc-STAT) has been cloned from the lepidopteran, Hyphantria cunea. The domain involved in DNA interaction and the SH2 domain are well conserved. The gene is transcribed at a low level during all stages of development, and the protein is present in hemocytes, fat body, midgut, epidermis, and Malphigian tuble (Mt). Especially, hemocytes and Mt showed transcriptional activation of Hc-STAT upon Gram (-) bacteria and fungal challenge. Gram (-) bacteria and fungal challenge specifically results in nuclear translocation of Hc-STAT protein and induction of DNA-binding activity that recognizes a STAT target site in H. cunea hemocyte. In vitro treatment with pervanadate translocates Hc-STAT to the nucleus in hemocyte cells. Here we report the first evidence for the involvement hemocyte JAK/STAT pathway upon microbial infection in lepidopteran insect.

The subfamily Hoplolaiminae included economically important plant parasitic nematodes and consisted of more than 400 species, all having the diagnostic characters of a strongly annulated cuticle and a large stylet. Among the Hoplolaiminae genera, the genus Hoplolaimus species include species such as H. columbus, and H. galeatus that cause serious damage to crops and turf grass in the Southeastern United States. Traditional identification of species has been approached by interspecific variation of phenotypic traits that rely on morphological and morphometric characters. However, these taxonomic criteria are sometimes not practical because of their limited ability to discriminate species among closely related groups due to overlapping of important taxonomic characters. The exact species identification is needed to control target nematode and also quarantine. Therefore, genetic studies for development of molecular diagnostics, population biology, and disease management are required. In recent years, many molecular diagnostic methods have been used for the identification of plant parasitic nematodes. Advanced molecular techniques have been used that test traditional identification methods. In our studies, Hoplolaimus species showed that high genetic divergence in rDNA sequence is combined with low morphological diversity. Based on genetic information, we developed multiplex PCR for H. columbus, H. galeatus, and H. magnistylus and successfully amplified mixed populations. In molecular phylogeny, the subfamily Hoplolaiminae is an important out‐group of the Heteroderidae, a notorious plant parasite nematode group. Molecular phylogeny of the Hoplolaiminae will help us understand of pathways of pathogenesis. In our phylogenetic analysis using D2 and D3 expansion segments of 28S gene, the molecular data supported morphological based taxonomic schemes. To reconstruct more reliable phylogenetic analysis, correct assignment of each nucleotide within multiple sequence alignment is an important step. Sequence alignments based on secondary structure have been proposed as new alternative methods to obtain this goal. We predicted the secondary structure of D2 and D3 domain using computational predictions method such as the minimization energy method and comparative sequence analysis (co‐variation). Predicted secondary structure included 18 species with two outgroup species, Globodera rostochiensis, Rotylenchulus reniformis. Consensus secondary structure was obtained from closely related and distantly related species. Phylogenetically informative characters were distributed in the stem region (86.7%). These results support the effectiveness of stem and loop regions for phylogenetic analysis of the Hoplolaiminae.

Members of the Vps (Vacuolar protein sorting) protein family involved in the formation of the retromer complex have been discovered in a variety of species such as yeast, mouse, and human. A mammalian retromer complex is composed of Vps26, Vps29, and Vps35 proteins and plays and important role in cation-independent mannose-6-phosphate receptor retrieval from the endosome to the trans-Golgi network. In this study, we have identified the full-length sequences of the retromer components of Vps26, Vps29, and Vps35 in micro pigs. The cDNA sequences of these retromer components have been determined and the result showed there is 99% homology among the component counterparts from mouse, micro pigs, and humans. In addition, the retromer complexes formed with hetero-components were found in the brain of micro pigs. Based on above results, we suggest mammalian Vps components are well conserved in micro pigs.

생물학적 자극통제 수단으로 활용하기 위한 돼지 웅성 페르몬성 분자를 탐색하고자 일련의 냄새 분자로서 2-(cyclo-hexyloxy) tetrahydrofurane 유도체들의 정량적인 구조와 수용체인 porcine odorant binding protein (pOBP)간의 결합 친화력 상수(p(Od)50)에 대한 비교 분자장 분석(CoMFA)을 실행하였다. 가장 양호한 CoMFA 모델 AIV (r2 cv.(q2)=0.886 및 r2 ncv.=0.984)은 기질 분자 내 입체 중심(chiral center)의 절대 배열이 C1(R),C2(S)인 분자를 atom based fit 방법으로 배열하였을 경우의 standard field와 indicator field가 조합된 CoMFA장의 조건에서 유도되었다. 이 CoMFA 모델은 입체장 40.8% 정전기장 14.6%및 소수성장 44.6%가 결합 친화력 상수에 영향을 미치는 요소임을 나타내었다. 등고도의 분석 결과로부터 효과적인 결합 친화력 냄새 분자를 수식하는 데 몇 가지 가치 있는 정보를 얻을 수 있었다.

CTA ester bonds in TG molecules were not attacked by pancreatic lipase and lipases produced by microbes such as Candida cylindracea, Chromobacterium viscosum, Geotricum candidium, Pseudomonas fluorescens, Rhizophus delemar, R. arrhizus and Mucor miehei. An aliquot of total TG of all the seed oils and each TG fraction of the oils collected from HPLC runs were deuterated prior to partial hydrolysis with Grignard reagent, because CTA molecule was destroyed with treatment of Grignard reagent. Deuterated TG (dTG) was hydrolyzed partially to a mixture of deuterated diacylglycerols (dDG), which were subsequently reacted with (S)-(+)-1-(1-naphthyl)ethyl isocyanate to derivatize into dDG-NEUs. Purified dDG-NEUs were resolved into 1, 3-, 1, 2- and 2, 3-dDG-NEU on silica columns in tandem of HPLC using a solvent of 0.4% propan-1-o1 (containing 2% water)-hexane. An aliquot of each dDG-NEU fraction was hydrolyzed and (fatty acid-PFB ester). These derivatives showed a diagnostic carboxylate ion, (M-1)-, as parent peak and a minor peak at m/z 196 (PFB-CH3)- on NICI mass spectra. In the mass spectra of the fatty acid-PFB esters of dTGs derived from the seed oils of T. kilirowii and M. charantia, peaks at m/z 285, 287, 289 and 317 were observed, which corresponded to (M-1)- of deuterized oleic acid (d2-C18:0), linoleic acid (d4-C18:0), punicic acid (d6-C18:0) and eicosamonoenoic acid (d2-C20:0), respectively. Fatty acid compositions of deuterized total TG of each oil measured by relative intensities of (M-1)- ion peaks were similar with those of intact TG of the oils by GLC. The composition of fatty acid-PFB esters of total dTG derived from the seed oils of T. kilirowii are as follows; C16:0, 4.6 mole % (4.8 mole %, intact TG by GLC), C18:0, 3.0 mole % (3.1 mole %), d2C18:0, 11.9 mole % (12.5 mole %, sum of C18:1Ω9 and C18:1Ω7), d4-C18:0, 39.3 mole % (38.9 mole %, sum of C18:2Ω6 and its isomer), d6-C18:0, 41.1 mole % (40.5 mole %, sum of C18:3 9c,11t,13c, C18:3 9c,11t,13r and C18:3 9t,11t,13c), d2-C20:0, 0.1 mole % (0.2 mole % of C20:1Ω9). In total dTG derived from the seed oils of M. charantia, the fatty acid components are C16:0, 1.5 mole % (1.8 mole %, intact TG by GLC), C18:0, 12.0 mole % (12.3 mole %), d2-C18:0, 16.9 mole % (17.4 mole %, sum of C18:1Ω9), d4-C18:0, 11.0 mole % (10.6 mole %, sum of C18:2Ω6), d6-C18:0, 58.6 mole % (57.5 mole %, sum of C18:3 9c,11t,13t and C18:3 9c,11t,13c). In the case of Aleurites fordii, C16:0; 2.2 mole % (2.4 mole %, intact TG by GLC), C18:0; 1.7 mole % (1.7 mole %), d2-C18:0; 5.5 mole % (5.4 mole %, sum of C18:1Ω9), d4-C18:0 ; 8.3 mole % (8.5 mole %, sum of C18:2Ω6), d6-C18:0; 82.0 mole % (81.2 mole %, sum of C18:3 9c,11t,13t and C18:3 9c,11t,13c). In the stereospecific analysis of fatty acid distribution in the TG species of the seed oils of T. kilirowii, C18:3 9c,11t,13r and C18:2Ω6 were mainly located at sn-2 and sn-3 position, while saturated acids were usually present at sn-1 position. And the major molecular species of (C18:2Ω6)(C18:3 9c,11t,13c)2 and (C18:1Ω9)(C18:2Ω6)(C18:3 9c,11t,13c) were predominantly composed of the stereoisomer of sn-1-C18:2Ω6, <..