IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

Japanese pine sawyer beetle, Monochamus alternatus Hope (Coleoptera: Cerambycidae) is considered as a serious pest in pine trees. To develop an eco-friendly strategy to manage this forest insect, we collected entomopathogenic fungi from Korean soil and assessed their virulence against the adults of the insect in laboratory conditions. As a result, two isolates with conidial suspension (1.0×107conidia/ml), showed 87% and 90% mortality 12 days after fungal treatment, respectively. We assessed the potential of the fungi-derived destruxin and protease as additives to the fungal isolates, and they showed insecticidal activity via feeding and spraying treatments. Finally, we produced fungal conidia in massive solid cultures and formulated wettable powders, and now studying optimal conditions of oil-based formulation with two isolates. Based on these results, we are evaluating the control efficacy of the fungal agents against M. alternatus in field conditions.

The beneficial effect of silicon (Si) in increasing salt stress tolerance has been observed in many plants, including the cereal crops rice, wheat, and barley. In this experiment, we examined the effect of Si on the survival and growth of torenia (Torenia fournieri L inden ex F oum) ‘ Duchess Blue and White’ cultured in vitro in the presence and absence of salt stress. Previous reports had suggested that torenia exhibited low salt tolerance. Shoot buds isolated from 16-day-old seedlings were cultured on Murashige and Skoog (MS) medium containing 0, 50, or 100 mM NaCl alone or in combination with 1.8 or 3.6 mM Si supplied as K2SiO3. Plant survival rate was significantly reduced by NaCl supplementation compared with the control. The survival rate significantly increased to 100% when 1.8 or 3.6 mM Si was added to the MS medium containing 50 mM NaCl. However, only 31% of plantlets survived when 1.8 mM Si was added to the culture medium containing 100 mM NaCl. Shoot and root lengths significantly decreased with increasing NaCl concentration in the culture medium, whereas addition of NaCl to the MS medium also significantly reduced fresh and dry weights. However, Si supplementation significantly increased fresh and dry weights under 50 mM NaCl, compared with the control. The greatest fresh and dry weights were recorded when shoot buds were cultured on MS medium containing 50 mM NaCl and 3.6 mM Si. The activities of the antioxidant-scavenging enzymes superoxide dismutase (SOD), ascorbate peroxidase (APX), and catalase (CAT), but not peroxidase (POD), were markedly higher in the presence of 50 mM NaCl than the activity of the control. When Si was added to the medium containing 50 mM NaCl, activities of SOD, POD, APX, and CAT decreased as compared with the 50 mM NaCl treatment. Thus, Si-mediated tolerance to NaCl stress was not due to increased activity of antioxidant enzymes. Although Si was not effective in increasing tolerance to high salt concentrations, such as 100 mM NaCl, the results suggested that Si supplementation could effectively enhance tolerance to 50 mM NaCl stress.

The objectives of the present study were to examine the antioxidant activity of fractions with different isoelectric points from salmon enzymatic hydrolysates and obtain peptide fractions of sufficient amounts with higher antioxidant peptide fraction, which could be applied to the food and animal model systems. The salmon enzymatic hydrolysates were fractionated on the basis of the amphoteric nature of sample peptides by preparative isoelectric focusing without toxic solvents and reagents, which is termed autofocusing. Acidic and basic fractions showed higher 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity than the other fractions. The basic fractions showed higher hydroxyl (OH) radical scavenging activity. The weak acidic and weak basic fractions showed higher oxygen radical absorbance capacity (ORAC) values than the acidic and basic fractions. The acidic fraction showed higher metal chelating activity than other fractions. The acidic fraction suppressed lipid oxidation in the cooked patties to a greater extent than other fractions. These results suggest that peptides fractions from salmon enzymatic hydrolysate are effective antioxidant, and that autofocusing could be useful to increase antioxidant activity for application to food and animal model systems.

벗초파리는 우리나라에서는 심각한 해충으로 인식되지 않아 발생 기간 및 패턴, 기주 범위 등의 생태적인 연구뿐만 아니라 모니터링을 위한 우수 유인제 및 트랩의 연구도 제한적으로 수행된 실정이다. 본 연구는 기존 연구 결과를 참조하여 사과식초 함유 유인제 2종과 화학적 루어 2종 그리고 트랩 2종에 대한 선발 시험을 수행하였다. 선발 시험은 사육중인 벗초파리를 이용한 choice test와 함께 수목원, 딸기 농장, 블루베리 농장에서의 6반복 유인력 검증 실험을 수행하였다. 유인력 비교 결과 ACV + wine의 포획량이 높았으며, 블루베리 농장 인근 결과값은 전체 조사 항목에서 통계적 유의성을 보였다 (P < 0.05). 트랩 선발 실험 결과 모든 실험 장소에서 Dreves 트랩이 높은 포획량을 보였다. 이상의 결과를 통해 ACV + wine과 Dreves 트랩 조합을 선발하였으며, 이를 이용하여 향후 지속적인 모니터링, 월동 발생 조사 등의 연구에 활용할 수 있을 것으로 생각된다.

The common bed bug, Cimex lectularius, possesses a cholinesterase expressed exclusively in the salivary gland (ClSChE). In this paper, we investigated the molecular structure, tissue distribution patterns, and biochemical properties of ClSChE and showed that ClSChE exists as a soluble monomeric form or a soluble dimeric form connected by a disulfide bridge. Immunohistochemical analysis confirmed that ClSChE was expressed in the epithelial cells of both the salivary gland and the duct. In addition, the secretion of monomeric ClSChE through the proboscis during feeding was detected by western blotting using a ClSChE-specific antibody. To predict the role of ClSChE injected into the tissue of an animal host, we analyzed the extent of sequestration and hydrolysis of acetylcholine (ACh)/choline (Ch) by ClSChE by ultra-performance liquid chromatography-tandem mass spectrometry. Kinetic analysis revealed that ClSChE possesses extremely low Km (high affinity to ACh) and Vmax values. These findings suggest that ClSChE functions as a sequestering enzyme specific to ACh (not to Ch) by having a very strong affinity to ACh but an extremely long turnover time.

To elucidate the effect of cellular phone electromagnetic wave (EMW) exposure on the developing cerebellar cortex of neonatal Sprague-Dawley rats, animals were exposed to cellular phone electromagnetic waves for 1 hr per day for 3 weeks. At the end of the experimental period, animals were sacrificed by cardiac perfusion, after which histological samples were prepared and observed microscopically. In the EMW exposure group, external granule cells were remained partially in the external granular layer without migrating into the internal granular layer. In addition, dark stained shrunken Purkinje cells with pyknotic nuclei increased and the outline of cells became irregular and showed degenerative signs, such as mitochondrial swelling and disrupted cristae. Moreover, the cisternae of rough endoplasmic reticula and Golgi complex were severely swollen. Bergmann glial cells adjacent to the dark stained Purkinje cells were swollen and cytoplasmic organelles were scant. Dark stained shrunken granule cells were also observed and the outline of cells was irregular. The results of the present study suggest that cellular phone EMW exposure to neonatal Sprague-Dawley rats leads to a partial delay of early migration of cerebellar cortical cells and degenerative changes in Purkinje cells, Bergmann glial cells and granule cells.

Selecting an appropriate antigen with optimal immunogenicity and physicochemical properties is a pivotal factor to develop a protein based subunit vaccine. Despite rapid progress in modern molecular cloning and recombinant protein technology, there remains a huge challenge for purifying and using protein antigens rich in hydrophobic domains, such as membrane associated proteins. To overcome current limitations using hydrophobic proteins as vaccine antigens, we adopted in silico analyses which included bioinformatic prediction and sequence-based protein 3D structure modeling, to develop a novel periodontitis subunit vaccine against the outer membrane protein FomA of Fusobacterium nucleatum. To generate an optimal antigen candidate, we predicted hydrophilicity and B cell epitope parameter by querying to web-based databases, and designed a truncated FomA (tFomA) candidate with better solubility and preserved B cell epitopes. The truncated recombinant protein was engineered to expose epitopes on the surface through simulating amino acid sequence-based 3D folding in aqueous environment. The recombinant tFomA was further expressed and purified, and its immunological properties were evaluated. In the mice intranasal vaccination study, tFomA significantly induced antigen-specific IgG and sIgA responses in both systemic and oral-mucosal compartments, respectively. Our results testify that intelligent in silico designing of antigens provide amenable vaccine epitopes from hard-to-manufacture hydrophobic domain rich microbial antigens.

Ixodid ticks are notorious as the vector of severe fever with thrombocytopenia syndrome virus (SFTSV) in Korea. To confirm the transovarial transmission of SFTSV, we collected blood-fed adult ticks from animals in SFTS case reported area of Jeju island in 2015. A total of 476 ticks were collected from four host animal species, horse (81.3%), dog (9.0%), roe deer (7.1%) and wild boar (2.5%). They were identified as 1 genus and 2 species, Haemaphysalis longicornis (87.8%) and H. flava (12.2%). Among them, 131 adults hatched their eggs successfully. The body weight of females was highly correlated with the number of egg masses laid (P < 0.05). We confirmed that one adult H. flava was infected with SFTSV and its infection rate was 0.77%. The eggs of the SFTSV-infected adults also were infected with SFTSV and its transovarial transmission was confirmed. This result suggests that the parent generation of H. flava could transmit SFTSV transovarially into their F1 progeny and H. flava might be one of major vectors of SFTSV in Korea.

미꾸라지에서의 Nitrofuran계 대사물질인3-amino-2-oxazolidone (AOZ), 5-morpholinomethyl-3-amino-2-oxazolidinone (AMOZ), 1-ammino-hydantoin (AHD)와 semicarbazide (SEM)의 잔류량을 검사하기 위해HPLC-MS/MS를 이용한 신속한 정량법이 개발되었다. 2-nitrobenzaldehyde (2-NBA)를 이용해 50℃에서 1시간 동안 산 가수분해와 유도체화 과정을 거친 뒤에, 액-액 분배로 정제와 추출을 하였다. 회수율은 음성시료에 3가지 농도 0.5, 1.0, 2.0 μg/kg의 표준액을 첨가하여 평가하였고 평균 회수율은 75.1-108.1% 이었다. 정밀성(%RSD)은 일내 8.7% 이하, 일간 8.5% 이하였다. 직선성은 NBAOZ는 0.2-20 μg/Kg, NBAMOZ는 0.8-20 μg/Kg, NBAHD는 0.2-20 μg/Kg, NBSEM 는 0.1-20 μg/Kg 범위에서 모두 상관계수 0.99이상이었다. 검출한계(LOD)는 NBAOZ 0.06 μg/Kg, NBAMOZ 0.24 μg/Kg, NBAHD 0.06 μg/Kg, NBSEM 0.03 μg/Kg이었고, 정량한계(LOQ)는 NBAOZ 0.2 μg/Kg, NBAMOZ 0.8 μg/Kg, NBAHD 0.2 μg/Kg, NBSEM 0.1 μg/Kg 이었다. 가수분해 및 유도체화 소요시간을 1시간으로 줄여 만든 신속 간편한 이 시험법이미꾸라지 중 nitrofuran metabolites잔류량 분석에 적합함을 확인할 수 있었다.

Recently, Bacillus thuringiensis (Bt) cry genes encoding insecticidal Cry proteins have been widely applied for the construction of transgenic crops resistant to insect pests. This study aimed to construct novel mutant cry1Ac genes for genetically modified crops with enhanced insecticidal activities. Using multi-site directed mutagenesis, 34 mutant cry1Ac genes were synthesized and converted at 24 amino acid residues, located on domain I (8 residues) and domain II (16 residues). These mutant genes were expressed as a fusion protein with polyhedrin using the baculovirus expression system. The expressed proteins were occluded into polyhedra and activated stably to 65 kDa by trypsin. Among these, Mut-N04, N06, and N16 showed high levels of insecticidal activites against larvae of Plutella xylostella, Spodoptera exigua, and Ostrinia furnacalis. Mut-N16, which showed the highest insecticidal activity, is expected to be a desirable cry gene for introduction into transgenic crops. This study could provide useful means to construct mutant cry genes with improved insecticidal activities and expanded host spectrum for transgenic crops.