식품에 존재하는 주요 비타민 B12는 hydroxycobalamin, adenosylcobalamin, methylcobalamin이며 cyanocobalamin은 시리얼과 같은 강화식품에 사용되는 합성형이다. 비타민 B12는 식품 중에 낮은 농도로 존재하기 때문에 추출 및 농축과정이 매우 중요하다. 본 연구에서는 과자와 시리얼 같은 탄수화물 식품 중의 비타민 B12 추출을 위해 α-amylase처리와 산성조건과 cyanide 존재시 열처리, immunoaffinity column(IAC)로 농축과 세척, HPLC/DAD로 분리 검출하였다. 분석법 수행특성지표들을 측정함으로 분석법을 검증하였다. 정확성과 정밀성은 AOAC 가이드라인을 충족시켰고, LOD 0.01 μg/100g과 LOQ 0.03 μg/100 g을 얻었다. 국제정도관리 시험에서 z-score 1.4로 분석수행이 우수함을 확인하였고 QC chart를 이용한 분석품질관리로 모든 분석이 관리하에 있음을 확인하였다. 검증된 분석법으로 과자류 12종과 시리얼 11종의 비타민 B12 함량을 분석하여 각각 0.02-0.26과 0.03-2.67 μg/100 g 범위를 얻 고 이는 신뢰도높은 데이터로 식품성분표에 활용 가능할 것이며 IAC-HPLC/DAD법이 탄수화물 식품의 비타민 B12 데이터베이스 구축에 활용될 수 있을 것으로 보인다.

Recently, researches of molecular biology for the identification of root-knot nematode (RKN) species have been reported in plant quarantine. In this study, applicable and reproducible method to extract high quality genomic DNA from single nematode (Meloidogyne spp.) was developed. Also, the modified method was verified by DNA manipulation techniques such as PCR amplification and cloning. Single juvenile was floated in a drop of water and digested with proteinase K for 24 h. After that, DNA was extracted by using distilled water as extraction buffer. PCR amplification was carried out with universal primers spanning the internal transcribed spacer (ITS) region to distinguish species. When using the existing DNA detection method, quantification results showed that 42.86% of the deposited DNA was extracted. Whereas the modified DNA extraction method was increased to 100%. When PCR products test the direct sequencing using the ITS rDNA primers, it was also identified as M. javanica, M. incognita, and M. hispanica. Based on the studies conducted, the application of this modified method would be useful and efficient on plant parasitic nematode molecular assay.

For ex-vivo diabetic control, the voltammetric diagnosis of glucose (GU) was conducted with a modified carbon nanotube paste electrode, using handheld analytical circuits. The optimum analytical conditions were attained within the 0.5-4.0 ug/L working range and at the 0.06 ug/L detection limit, which system was interfaced to the feedback circuits and was applied to human urine for diabetic-patient diagnosis. It can be used for ex-vivo flow control analysis, vascular flow detection, and other medicinal assays.

Bean bug, Riptortus clavatus Thunberg (Heteroptera: Alydidae) causes serious damage to Leguminosae. Herein an entomopathogenic fungal virulence assay system against bean bugs was established to construct a fungal database which can be used in integrated pest management (IPM). First to obtain as many bean bugs as possible at the same stage, host plant-preference and developmental synchronization of bean bugs were investigated. In the preference assay, five pairs of adults were infested in a plastic cage, where a pot of green bean, pea or cowpea was previously placed. The highest fecundity and the fastest development of bean bug was observed in the green bean cage. Secondly, in the synchronization experiment, eggs were collected from the cage of adults in 1, 3, 5 and 7 days after oviposition and transferred to a fresh cage with green beans. From the every 4 days of survey, similar stages of bean bugs were found in the cages with the oviposition for 1 and 3 days, rather than the longer times of oviposition. A fungal bioassay against bean bugs was conducted using the bean bugs from the above insect rearing system. Ten Beauveria bassiana isolates were cultured on quarter-strength Sabouraud dextrose agar (¼SDA) for 7 days at 25°C. Ten 4th instar of nymphs were placed on a cultured plate for 1 hour and tranferred to a fresh moisturized plate with grains of green bean. ERL836 isolate treatment showed the highest virulence and fungal mycosis was observed on the bean bugs. In conclusion, these results can be useful to establish an entomopathogenic fungal database for IPM.

Cronobacter muytjensii는 유아용 조제분유(IFP)의 잠재적 위험요인으로 중요한 식품 기인성 병원균이다. 이 연구에 서는 C. muytjensii 검지를 위한 특이적 면역글로불린 G(IgG)를 개발하고, 이 anti-C. muytjensii IgG를 이용하여 간접 비경쟁 효소면역측정법(INC-ELISA)을 개발하였다. 그 결과, 새롭게 개발한 INC-ELISA 방법은 C. muytjensii에 매우 민감하고, 순수배양 시 6.5×103 CFU/㎖의 검출한계와 유아용 조제분유에서 1 cell/25 g의 검출한계를 나타내었다. INC-ELISA 방법은 또한 C. muytjensii에 탁월한 특이성을 보이고, Cronobacter 속 외 11종의 다른 식품 기인성 병원균 계통과의 교차반응을 보이지 않았다. 이러한 결과는, 개발된 INC-ELISA 방법이 C. muytjensii에 매우 민감하고 효율적 이며, 신속하고 용이한 검출을 위한 진단 키트 개발에 적용할 수 있음을 시사한다.

Beet armyworm, Spodoptera exigua are difficult to control using chemical insecticides because of the development of insecticide resistance. For eco-friendly beet armyworm managements, various control agents are required. Entomopathogenic fungus is one of promise control agents as an alternative to chemical control agent. We isolated entomopathogenic fungi from soil samples of suwon by insect-bait method using Galleria mellonella and conducted bioassay to larva of beet armyworm. As a result of bioassay isolate FG274, FG340, FG344 had high virulence as 100% against second instar larva of S. exigua. To identify the fungus isolates, their’s morphological characteristic was observed and ITS of 18srRNA was sequenced. ITS sequence of FT274, 340, 344 were highly matched (100%) to that of Beauveria bassiana, Paecilomyces fumosoroseus, Metarhizium anisopliae. To investigate the optimal concentration, three isolates were sprayed at three different concentration(1×106 ,107 and 108 conidia/㎖) in laboratory conditions. 나타내었다.

As veterinary medicine industry has grown up steadily, the government quality control of veterinary medicine in market has been performed since 1964. The result of post-market surveillance assay from 2009 to 2012 was analyzed in this study. The average violation rate of all veterinary medicines, was 3.2%, 2.8%, 2.8% and 1.4% in 2009, 2010, 2011 and 2012, respectively. The total average violation rate in the category of antibiotics, other drugs except antibiotics, biologics were 1.2%, 4.4% and 0.8%, respectively. The violations due to insufficiency of major active ingredient content occupied 76.7% among the total violations. The most frequent violated in antibiotics were ampicillin and tylosin, followed by sulfonamides, penicillins etc. In case of other drugs except antibiotics, vitamin A and glucocorticoids (dexamethasone and prednisolone) were violated frequently. The overall trend of violation rates gradually decreased from 2009 to 2012, suggesting that quality of veterinary medicines has improved according to Korea Veterinary Good Manufacturing Practice (KVGMP) system settled down in 1988.

최근 성인병에 대한 대처방안으로 천연식품들 중에서 기능성 물질을 섭취하려는 요구 증가 와 함께 생리활성물질의 개발에 노력하고 있으며, 그중에서도 질병의 치료 및 예방에 효과가 있는 천연물로 버섯이 주목 받고 있다. 그 중 약용버섯은 항염․항암작용, 항산화 활성 효과 등이 보고되고 있으며 다양한 연구가 많이 이루어지고 있다. 본 실험에서는 영지버섯, 노루궁 뎅이버섯, 상황버섯, 동충하초 자실체를 70% 에탄올로 추출하여 아질산염소거능과 Raw 264.7 세포에서 NO 생성 저해 효과를 알아보기 위하여 연구가 진행되었다. 아질산염은 2급 및 3급 amine류와 반응하여 발암물질인 Nitrosoamine을 생성하는 것으로 알려져 있다. Nitrosoamine 은 체내에서 diazoalkane(CnH2nN2)으로 변화하여 핵산이나 단백질 또는 세포내의 성분을 알 카리화 함으로써 암을 유발한다고 알려져 있다. 아질산염 소거능(nitrite-scavenging effect)은 Gray등의 방법으로 520nm에서 흡광도를 측정하여 잔존하는 아질산염량을 구한 결과 대조구 Ascorbic acid에 반해 영지버섯 균주 중 ASI 7002와 상황버섯은 보다 높은 소거능을 보였다. NO(nitrite oxide)형성은 체내방어기능, 신호전달기능, 혈관확장 등의 다양한 생리기능을 가 지고 있지만 과도한 NO의 형성은 염증을 심화시켜 조직의 손상, 유전자변이, 패혈성 쇼크 및 신경손상 등을 일으킨다. Raw 264.7 대식세포에 LPS(lipopolysaccharides) 자극에 많은 양의 NO를 형성하며 이에 의한 세포 독성은 염증반응 관여 정도를 알아본 결과 영지버섯 균주 중 ASI7002와 ASI7063이 대조구 Ascorbic acid보다 높은 결과를 보여줬고, 이와 더불어 염증관 련 단백질인 iNOS, COX-2 발현을 억제하는지 여부를 RT-PCR을 통해 확인한 결과, 열수추 출한 영지버섯 균주 ASI 7002에서 농도의존적으로 억제함을 알 수 있었다.

The electrochemical analysis of silver ion was performed using cyclic voltammetry (CV) and square-wave (SW) stripping voltammetry, and electrode cell systems were fabricated with graphite pencil electrode (GE) of working, reference and counter electrodes. Also electrolyte was the use of sea water as electrolyte solutions instead of ionic controlled solutions. The optimum analytical conditions for the cyclic and stripping parameters were determined using GE. The results approached the microgram working ranges of SW(ug/L) and CV(ug/L) Ag, and the optimum conditions were applied to frog's tissue and the food samples.

Diagnosis with an ex-vivo gold sensor was done using a modified fluorine-doping sensor, and cyclic voltammetry (CV) redox potentials of 0.4 V anodic and -0.2 V cathodic were obtained. Both peak currents were optimized using square-wave (SW) stripping voltammetry, and an analytical working range of 10-80 ug/L SW was attained. The precision of the 10-mg/L Au was 0.765 (n=8) RSD under the optimum conditions, and the analytical detection limit approached 0.006 ug/L (S/N=3) with only a 60 sec accumulation time. The developed method was used to examine the mouse droppings for medicinal diagnosis.

In this study, a rapid immuno-chromatographic assay was developed to detect Salmonella Typhimurium. This assay was based on the lateral flow immunoassay. Two different anti-Salmonella polyclonal antibody (Pab) are used in this strip assay for the double antibody sandwich immunoassay method. The strip sensor was composed of sample pad, conjugation pad, membrane, and absorbent pad. For the optimal conjugation between nano-gold and anti-Salmonella Pab, the pH value of colloidal gold solution was determined to 8.4. To prevent non-specific binding between nano gold and anti-Salmonella Pab, sample pad and membrane was treated with PBS buffer, included 2% BSA and 10% sucrose. The control and detection zones were visually detected according to antigen-antibody binding. The detection limit of the developed strip assay was 105 CFU/mL within 10 min, and improved until 104 CFU/mL after 12 hr.

A investigation of electrochemical analysis of antibiotics Neomycin (C23H46N6O13) was searched using electrochemical square wave (SW) stripping and cyclic voltammetry (CV) using working sensor of the modified carbon nanotube combination electrodes, optimum diagnostic parameters were searched by anodic stripping, final conditions were attained to working range of 1.0-14.0 ng/L, detection limit (S/N) was found to be 0.6 ng/L. The developed method was discovered to be fitting in quality control in the food, pharmaceutical and other manufacturing sectors.

This study was carried out to apply an optimized convenient assay, exploiting azo dye-bound chromogenic substrates, to measurement of protease activity. When determined for responses at varying concentrations of two substrates, azocasein and azoalbumin, using 0.5 and 5.0 mg/mL each of bovine pancreas trypsin, 3% azocasein was found to be the most appropriate substrate solution to measure protease activity. Compared with a conventional casein-Folin phenol assay, the chromogen-based protease assay exploiting 3% azocasein showed better precision to have a coefficient of variability in seven repetitive measurements less than 1.11%. When various reagent-grade and industrial proteases that showed proteinase or peptidase activities were tested by this assay at increasing enzyme concentrations, typical shape of rectangular hyperbola in activity-enzyme concentration profiles was observed. In addition, the assay of this study was suitable for activity measurement in real samples that were prepared by hydrolyzing wheat gluten and anchovy fine powder with proteases.

This study was performed to develop and optimize a rapid strip kit for the dual-detection of Aflatoxin B1 (AF) and Ochratoxin A (OTA). The strip kit is composed of sample pad, conjugation pad, nitrocellulose membrane, and absorbent pad. Manually spotted mycotoxin-BSA conjugates and anti-mouse IgG on nitrocellulose membrane were used as the test line and the control line, respectively. Conditions for rapid and easy detection of mycotoxins, conjugation between toxin-MAbs and nano Au particles, pretreatment method of pads, and composition of standards solutions were optimized. Feasibility of the strip kit to detect AF and OTA were evaluated with standard samples. Test results were acquired in 10 min and the detection limits were 100 ppm for AF and OTA, respectively.

Bio assay of mercury and cadmium ions were searched using voltammetric analysis using DNA doped carbon nanotube paste electrodes (DCP). The square-wave stripping voltammetryic optimized results indicated working ranges of 1-10.0 ngL-1 and 20-100 ugL-1, Hg(II) Cd(II) within an accumulation time of 120 seconds, in 0.1-M phosphate buffer solutions of pH 6.3. The relative standard deviations of 5 ngL-1 Hg(II) and Cd(II) that observed were 0.14 and 0.22% (n=12), respectively, using optimum conditions. The low detection limit (S/N) was pegged at 0.1 ngL-1 (4.9×10-11M) Hg(II) and 0.2 ngL-1 (1.77×10-10M) Cd(II). The developed methods can be applied to assays in biological fish kidneys and water samples.

한국원자력연구원 내 저장중인 중·저준위방사성폐기물의 안전한 영구처분을 위하여 방사성폐기물처 리시설에서는 이들 저장폐기물의 핵종재고량 평가를 위한 드럼핵종분석장치의 도입을 기획하고 있으며, 이 장치를 운영하고 유지보수하기 위한 전용의 평가시설 구축을 추진하고 있다. 이 평가시설 외부에는 제 1방사성폐기물저장시설이 인접하여 있으며 내부에는 평가대상드럼들과 드럼핵종분석장치의 밀도보정용 선원 등이 존재함으로 이들이 방사선원항으로 작용하여 평가시설 내·외부의 선량률에 영향을 미치게 된 다. 따라서 방사선원항 주변의 콘크리트 구조물에 대한 방사선차폐 영향을 평가하여야 한다. 본 연구에서 는 MCNP 코드를 이용하여 해당 방사선원항으로부터 평가시설 내부 방사선관리구역을 둘러싸고 있는 콘 크리트 구조물에 대한 방사선 안전성을 평가하였다. 평가결과 현재 고려되고 있는 콘크리트 구조물의 약 30 cm 두께는 해당 방사선원항으로부터의 방사선을 차폐하기에 충분한 것으로 확인되었다.

Vollenhovia emeryi ant is distinguished by its wing morphology; short winged (SW) and long winged (LW). Its reproduction shows a bizarre genetic caste system distinct from other social hymenopteran insects. Unfertilized eggs undergo genome duplication and develop into clonal gynes. Fertilized eggs develop either into workers or males. The fate of the fertilized eggs is determined whether maternal genome loss (MGL) takes place after fertilization. Eggs with MGL become haploid males with only paternal half of the genome. Without MGL, the eggs become workers with maternal and paternal half of the genome. In this research, we analysed 5 nuclear genes of SW and LW individual ants. Among them, two genes from an SW male are identical to those of LW, and one gene from the SW male seems a variant of LW. The result indicates that SW males are derived from LW colonies. From the genetic relatedness point of view individuals in the same castes are genetically identical. On the other hand, between workers and two reproductives, the relatedness is asymmetrical and there is even no gene sharing between gynes and males. The conventional genetic relatedness by Hamilton is revised under this condition.