Mealworms, Tenebrio molitor (L.) is used as an important animal feed additive for growth promotion and health management, but potentially exposes to fungal infection. In this work, virulence of two species of entomopathogenic fungi against the insect, and the relationship between abiotic features and virulence were investigated. Secondly our consideration was also given to the effect of chemical fungicides on conidial germination for risk control. Between Beauveria bassiana (Bb) and Metarhizium roberstii (Mr) (previously M. anisopliae), Bb isolates had much higher virulence (~100% mortality in 3~4 days after the treatment), rather than Mr isolates in laboratory assays. Next, fungus-treated mealworms were kept at wheat bran at 20, 25, 30 and 35℃ with 3, 6, 9 times of water spray to the feeds for set-up of different humidity conditions. Inoculation of fungi to mealworms was conducted by fungal spray and feeding methods, which resulted in higher virulence in feeding method. In the feeding method, all temperature treatments except 35℃ showed high virulence against mealworms, but any significant relationship between virulence and humidity was not observed. In the chemical fungicide screening, fluazinam (CAS No. 79622-59-6) and mancozeb (8018-01-7) significantly inhibited the germination of Bb and Mr conidia. This work suggest that contamination of wheat bran with fungal pathogens, particularly B. bassiana may induce mycosis of mealworms, but introduction of effective fungicides possibly reduce fungal infection.

Beauveria bassiana isolates have been used in integrated pest management, but little consideration has been given to the studies on fungal gene expressions and their functions. In this work, to determine the functions of genes, B. bassiana ERL1170 was transformed by restriction enzyme-mediated integration method, where pABeG with bar gene was used as a transformation vector. Among seven hundred of transformants, morphologically different ERL1170-pABeG-#160 transformant, particularly dysfunctional in conidiogenesis. The transformant had yellow hyphal growth on fourth strength Sabouraud dextrose agar (SDA/4) and produced very small amount of conidia (<1.0×105 conidia/cm2 agar) in 7 days, whereas wild type had white mycelial growth and significantly greater conidia (3.6×106 conidia/cm2 agar). Additionally under microscopic observation, hyphae of #160 seemed like indian club, compared to the straight forms of wild type hyphae. The next work is figure out possible genes contributing the conidiogenesis of B. bassiana.

Entomopathogenic fungus, Beauveria bassiana can be used in integrated crop management and pharmaceutical applications. Recently some efforts have been given to the fungus to improve its biological performances, but low fungal transformation efficiency is one of the limitations. In this work, B. bassiana ERL1170 isolate was used for fungal transformation by restriction enzyme-mediated integration, where pBARKS1-Bbs-cecropinA was linearied using HindIII. The fungal transformation comprised two steps, preparation of competent blastospores and integration of the plasmid into the cells. To prepare competent blastospores, 2-d cultured blastospores were individually treated with 0.1, 0.2, 0.4 and 0.8 M lithium acetate (LiAc). Secondly in the integration step, concentration of LiAc and calcium chloride (CaCl2), and time period of heat shock were investigated as follows: LiAc, 1, 2 and 4 M; CaCl2, 50, 100 and 200 mM; and heat shock at 42℃, 20, 40 and 60 min. Consequently, combination of 0.2 M LiAc in preparing competent blastospores and 2 M LiAc and 200 mM CaCl2 in the second step showed the highest transformation efficiency. This work would be helpful in the fungal transformation of B. bassiana.

Poly(vinyl chloride)-g-poly(oxyethylene methacrylate) (PVC-g-POEM) 가지형 공중합체를 원자전달라디칼 중합을 통해 합성하여 전기변색소자의 전해질에 적용하였다. 가소화된 고분자 전해질은 가소제로서 propylene carbonate (PC)/ethylene carbonate (EC) 혼합물을 도입하여 제조하였으며, Lithium tetrafluoroborate (LiBF4), lithium perchlorate (LiCIO4), lithium iodide (LiI) and lithium bistrifluoromethanesulfonimide (LiTFSI)를 사용하여 염의 종류에 따른 영향을 조사하였다. 광각 x-선 산란(WAXS)과 시차주사 열량법(DSC) 측정 결과 고분자 전해질의 구조와 유리전이온도(Tg)가 변하였고, 이는 POEM 내의 에테르의 산소와 리튬염 사이의 상호작용으로 인해 변했다는 것을 FT-IR 분광법을 통하여 확인하였다. 투과전자현미경(TEM) 측정 결과 PVC-g-POEM 가지형 공중합체의 미세상분리 구조가 PC/EC와 리튬염의 도입에도 변하지 않는 것을 관찰하였다. 가소화된 고분자 전해질은 poly(3-hexylthiophene) (P3HT) 전도성 고분자를 이용한 전기변색소자에 적용되었다.

원자전달 라디칼 중합(ATRP)에 의해 poly(vinyl chloride) (PVC) 주사슬과 poly(styrene sulfonic acid) (PSSA) 곁사슬로 되어있는 양쪽성 PVC-g-PSSA 가지형 공중합체를 합성하였다. PVC-g-PSSA 가지형 공중합체 고분자를 템플레이트로 사용하고 졸겔법을 적용하여, 결정성 아타네제상의 미세기공 이산화티타튬 필름을 제조하였다. TiO2 전구체인 TTIP를 친수성인 PSSA 영역과 선택적으로 작용시켜 TiO2 메조기공 필름을 성장하였으며, 이를 주사전자 현미경 (SEM)과 엑스레이회절 (XRD)분석을 통해 분석하였다. 스핀코팅 횟수와 P25 도입에 따른 염료감응 태양전지 성능을 체계적으로 분석하였다. 그 결과 준고체 고분자 전해질을 이용하였을 때, 100 mW/㎠ 조건에서 에너지 변환 효율이 2.7%에 이르렀다.

원자전달 라디칼 중합(ATRP)에 의해 poly(vinyl chloride) (PVC) 주사슬과 poly(styrene sulfonic acid) (PSSA) 곁사슬로 되어있는 양쪽성 PVC-g-PSSA 가지형 공중합체를 합성하였다. 합성된 고분자 전해질막을 10 wt% AgNO3 수용액에 담가 은이온으로 이온교환을 하였으며, 환원제를 통하여 은 나노입자를 성장시켰다. UV분광학과 XRD 분석을 통해 은 나노입자 성장을 확인하였다. 투과전자현미경(TEM) 분석결과 NaBH4를 사용하였을 때 10~20 nm 크기의 은 나노입자를 얻는데 가장 효과적임을 알 수 있었다. 또한 은 나노입자의 성장은 환원제의 농도와 환원 시간에 크게 영향을 받았다.

This study was conducted in order to investigate repeated-dose toxicities of Magnolia ovobata ethanol extract (MEE). MEE was administered orally to male and female Sprague Dawley rats at dose levels of 0, 500, 1,000, or 2,000 mg/kg for four weeks. Repeated administration of MEE did not induce abnormalities in general signs, body weight gain, feed and water consumption, necropsy findings, or organ weights. In addition, no abnormality was observed in hematological analyses; red blood cells and their indices, white blood cells, platelets, and coagulation times. In male rats, BUN and creatinine showed an increase at doses of 2,000 mg/kg and 500-1,000 mg/kg, respectively, while in female rats, lactate dehydrogenase and creatine phosphokinase showed a decrease at 2,000 mg/kg, the upper-limit dose of repeated-dose toxicity studies. However, there were no dose-dependent increases or gender-relationship. In addition, other parameters of the hepatic and muscular toxicities as well as energy and lipid metabolism were not affected. In microscopic examination, no considerable pathological findings were observed. The results indicate the safety of oral administration of MEE to the upper-limit dose.

Poly(vinyl chloride)-g-poly(styrene sulfonic acid) (PVC-g-PSSA) 가지형 공중합체를 합성한 후, 이를 이용하여 80℃에서 열적으로 환원하여 은 나노입자를 제조하였다. 반응 시간을 바꿈에 따라 다양한 구조의 은 나노입자를 제조하는데 성공하였다. 1시간 정도의 짧은 반응 시간에서는 가지형 공중합체의 미세 상분리 구조를 크게 변화시키지 않고 5 nm 크기의 작은 은 나노입자가 생성되었다. 5시간 정도의 중간 반응 시간에서는 30 내지 50 nm 정도의 크기를 갖는 은 나노입자가 생성되었다. 18시간 정도의 긴 반응 시간에서는, 은입자가 뭉친 허리케인 모양의 은 집합체가 관찰되었다.

Previous studies indicated that Matsumuraeses phaseoli and M. falcana (Lepidoptera: Tortricidae) are separate species since a few differences were observed in genitalia morphology and female sex pheromone composition. A clear difference was detected in the DNA sequences of cytochrome oxidase I of the two species separately collected in different plants and regions. A hybridization test also showed that a post-zygotic reproductive isolation occurred between the species. In field monitoring, however, both species have been caught simultaneously and together in the separate sex pheromone traps installed for the two species around neighboring soybean and red bean fields. Molecular marker-assisted identification with several adults sampled from the trapped insects showed that only ca. 40% of M. phaseoli adults identified as the species by genitalia morphology was the M. phaseoli, while ca. 97% of M. falcana adults identified as the species was the M. falcana. The result indicated that the observation of genitalia did not make a decisive criterion for classification of the insects. Conclusively, it suggested that the sex pheromones of the two species should be studied more precisely although there is a possibility that the two species are hybridized in fields as in laboratory, and speciation is under process.

The main nutritional problem of sorghum×sudangrass (sudangrass) silage is low quality and dry matter (DM) contents. This experiment was conducted to evaluate whether agricultural byproducts addition of sorghum×sudangrass silage increases DM content and forage quality of the silage. Sorghum×sudangrass silage with added byproducts had low pH values, however sorghum×sudangrass silage (control) had a high pH value because of its high moisture content. Silage added with byproduct had lower acid detergent fiber (ADF) and neutral detergent fiber (NDF) contents than control silages, while its non-fiber carbohydrate (NFC), and total digestible nutrients (TDN) and lactic acid contents showed the opposite results. Silages added with crushed rice and green grain of rice were higher in lactic acid content than othor treatments as well as high DM, NFC, and TDN contents. Silage added with crushed rice and green grain of rice can be recommended as the most effective treatment for increasing forage quality and DM content of sorghum×sudangrass silage.

To find some antibacterial peptides responsible for bacterial resistance, we performed differential hybridization with total cDNA probes which synthesized from normal and immunized larvae. Thirteen individual cDNA transcripts were expressed differentially in a total 1,862 random cDNA clones. One of upregulated genes is a novel member of the insect defensin-like peptide(Coprisin), a family of antibacterial peptide. Northern blot analysis showed that Coprisin was up-regulated at 4h and reached the highest point level at 16h after injection of E.coli. The deduced amino acid sequence of Coprisin was composed of 80 amino acids with predicted molecular weight of 8.6 kDa and PI of 8.72. Comparison of the deduced amino acid mature portion of Coprisin with defensin-like peptide of other insect indicated that it has 79.1% and 67.4% identity with Anomala cuprea and Allomyrina dichotoma, respectively. To find antibacterial active region of Coprisin, we synthesized four peptides corresponding to amino acid residues 1V-43N-NH2(CopN1), 5-16(CopN2), 19-30(CopN3) and 31-43(CopN4) of coprisin having amidated amino acid residues at their Cterminal. A 12-mer amidated at its C-terminus, ACALHCIALRKK-NH2 (Ala19-Lys30-NH2) was synthesized based on the deduced amino acid sequence, assumed to be an active site sequence. This peptides showed antibacterial activity against E.coli, Staphylococcus aureus, MRSA, Psedomonas syringae, and Pectobacterium carotovorium. Modified 9-mer peptide, LRCIALRKK-NH2, showed strong antibacterial activity than mellitin peptide used as a positive control against gram-negative and gram-positive bacteria. This peptide showed no haemolytic activity and quite stable at 100℃ for several hours of incubation and in a wide pH range(pH2-12). Therefore, this peptide may be a good candidate for the development of new drug with potent antibacterial activity without cytotoxicity.

Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, H+-transporting ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.