• Archives of Pharmacal Research
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• 제28권11호
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• pp.1293-1301
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• 2005

Flavonoids, a group of low molecular weight phenylbenzopyrones, have various pharmacological properties including antioxidant, anticancer, bactericidal, and anti-inflammatory. We carried out anti-herpetic assays on 18 flavonoids in five classes and a virus-induced cytopathic effect (CPE) inhibitory assay, plaque reduction assay, and yield reduction assay were performed. When flavonoids were applied at various concentrations to Vero cells infected by HSV-1 and 2, most of the f1avonoids showed inhibitory effects on virus-induced CPE. Among the flavonoids, EC, ECG (flavanols), genistein (isoflavone), naringenin (flavanone), and quercetin (flavonol) showed a high level of CPE inhibitory activity. The antiviral activity of flavonoids were also examined by a plaque reduction assay. EC, ECG, galangin, and kaempferol showed a strong antiviral activity, and catechin, EGC, EGCG, naringenin, chrysin, baicalin, fisetin, myricetin, quercetin, and genistein showed moderate inhibitory effects against HSV-1. In these experiments, flavanols and flavonols appeared to be more active than flavones. Furthermore, treatment of Vero cells with ECG and galangin (which previously showed strong antiviral activities) before virus adsorption led to a slight enhancement of inhibition as determined by a yield reduction assay, indicating that an intracellular effect may also be involved.

• The Plant Pathology Journal
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• 제33권2호
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• pp.184-192
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• 2017

Tylenchulus semipenetrans is an important and widespread plant-parasitic nematode of citrus worldwide and can cause citrus slow decline disease leading to significant reduction in tree growth and yield. Rapid and accurate detection of T. semipenetrans in soil is important for the disease forecasting and management. In this study, a loop-mediated isothermal amplification (LAMP) assay was developed to detect T. semipenetrans using DNA extracted from soil. A set of five primers was designed from the internal transcribed spacer region (ITS1) of rDNA, and was highly specific to T. semipenetrans. The LAMP reaction was performed at $63^{\circ}C$ for 60 min. The LAMP product was visualized directly in one reaction tube by adding SYBR Green I. The detection limit of the LAMP assay was $10^{-2}J2/0.5g$ of soil, which was 10 times more sensitive than conventional PCR ($10^{-1}J2/0.5g$ of soil). Examination of 24 field soil samples revealed that the LAMP assay was applicable to a range of soils infested naturally with T. semipenetrans, and the total assay time was less than 2.5 h. These results indicated that the developed LAMP assay is a simple, rapid, sensitive, specific and accurate technique for detection of T. semipenetrans in field soil, and contributes to the effective management of citrus slow decline disease.

• 한국자원식물학회지
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• 제29권6호
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• pp.704-711
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• 2016

Tomatoes are considered as one of the main components of daily meals in most households. Thus, it is important to invest in studies enhancing their yield and nutritional value. The study evaluated the effect of wood vinegar (WV) on tomato under greenhouse conditions. Data on fruit number, fruit weight, and plant height were recorded. Total phenolic content (TPC) and antioxidant activity of tomato were determined using the Folin-Ciocalteu reagent and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging assay, respectively. FRR-CF+500x-WV significantly increased the fruit number (86.11%) and fruit weight (81.78%) over the control. Results were comparable to HRR-CF+250x-WV, indicating that conventional fertilizer and WV may have synergistic effect on each other. TPC and scavenging effect on DPPH radical was improved by 250x-WV application. The increase in the phenolic compounds can be attributed to WV as a direct source or as a factor triggering the plants to produce more secondary metabolites with the purpose of increasing natural defenses. The significant effect obtained by applying the full recommended rate of conventional fertilizer on the carotenoid content was due to the availability of the major nutrients needed by the plant. Correlation analysis showed that carotenoids have negative correlation with TPC and DPPH. However, TPC and DPPH showed that these are positively correlated with each other. Tomatoes are excellent source of antioxidants associated with the reduction of some human diseases and improved health. The results provided evidence that WV alone and/or its combination with conventional fertilizers has favorable effects on the quality of tomato.

• 한국작물학회지
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• 제48권6호
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• pp.501-505
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• 2003

보리호위축병 피해지역에 대한 수량감소의 예측을 위하여, 보리호위축병(BaYMV)에 대한 저항성 품종과 이병성 품종간 수량구성요소, 수량 및 증질 변이를 건전구와 이병구에서 비교 조사하였다. 1. 보리호위축병의 이병주는 BaYMV에 감염된 것으로 나타났으며, 이병정도는 감수성 품종인 백동에서 9, 저항성 품종인 내한쌀보리에서 1정도 발병하였다. 2. 감수성 품종인 백동은 천립중을 제외한 간장, 수당립수, $\textrm{m}^2$당 수수 등의 수량구성요소가 유의하게 감소하였으며, 수량은 대조구의 20%수준으로 현저히 감소하였다. 3. 보리호위축병 감염구에서 종자의 발아율은 감수성 품종과 저항성 품종간에 차이가 없었으나, 정립률은 감수성 품종인 백동에서 설립중이 유의하게 증가하였으며, 대조구에 비하여 정립률과 조단백질 함량 모두 유의한 차이를 보였다. 4. 보리호위축병은 간장, 수량, 정립률, 수수, 천립중, 1수립수 등의 피해와 부의 상관을, 조단백질 함량과는 정의 상관을 보여 보리호위축병에 이병이 되면 수량 및 품질에 크게 피해를 주는 것으로 나타났다.

• Journal of Microbiology and Biotechnology
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• 제13권5호
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• pp.731-737
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• 2003

Mucinase complex from V. parahaemolyticus ATCC 17802 was purified 6-fold with 0.4% yield by two sequential steps of Q-Sepharose and Superdex 200HR column chromatographies. Partially purified mucinase complex showed at least 8 times higher mucin-degrading activity than the culture filtrates. The mucinase complex also showed gelatin-and-casein-hydrolyzing activities, which demonstrates that the protein is a complex compound containing several proteases. The optimum pH and temperature of partially purified mucinase complex for mucin degradation was 8.0 and $35^{\circ}C$, respectively. The partially purified mucinase complex showed high cytotoxic activity on vero cells when examined by MTT assay and microscopic observations. Cytotoxicity was significantly increased in proportion to the concentration of the mucinase complex. Mouse experiments revealed that the jejunum, ileum. and large intestinal tissues were damaged by the injection of the mucinase complex. In particular, the reduction of the goblet cells in the large intestine was remarkable. Collectively, these data suggest that the mucinase complex partially purified from V. parahaemolyticus ATCC 17802 contributes to the adhesion and invasion of V. parahaemolyticus into the host intestinal tract.

• Journal of Microbiology and Biotechnology
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• 제22권3호
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• pp.316-325
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• 2012

Xylanase has been used extensively in the industrial and agricultural fields. However, the low-yield production of xylanase from native species cannot meet the increasing demand of the market. Therefore, improving the heterologous expression of xylanase through basic gene optimization may help to overcome the shortage. In this study, we synthesized a high-GC-content native sequence of the thermostable xylanase gene xynB from Streptomyces olivaceoviridis A1 and, also designed a slightly AT-biased sequence with codons completely optimized to be favorable to Pichia pastoris. The comparison of the sequences' expression efficiencies in P. pastoris X33 was determined through the detection of single-copy-number integrants, which were quantified using qPCR. Surprisingly, the high GC content did not appear to be detrimental to the heterologous expression of xynB in yeast, whereas the optimized sequence, with its extremely skewed codon usage, exhibited more abundant accumulation of synthesized recombinant proteins in the yeast cell, but an approximately 30% reduction of the secretion level, deduced from the enzymatic activity assay. In this study, we developed a more accurate method for comparing the expression levels of individual yeast transformants. Moreover, our results provide a practical example for further investigation of what constitutes a rational design strategy for a heterologously expressed and secreted protein.

• 대한약침학회지
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• 제14권3호
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• pp.19-45
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• 2011

Objectives: This study was performed to analyse the effects of Sweet Bee Venom(Sweet BV-pure melittin, the major component of honey bee venom) on the central nervous system in rats. Methods: All experiments were conducted at Biotoxtech Company, a non-clinical studies authorized institution, under the regulations of Good Laboratory Practice (GLP). Male rats of 5 weeks old were chosen for this study and after confirming condition of rats was stable, Sweet BV was administered in thigh muscle of rats. And checked the effects of Sweet BV on the central nervous system using the functional observational battery (FOB), which is a neuro-toxicity screening assay composed of 30 descriptive, scalar, binary, and continuous endpoints. And home cage observations, home cage removal and handling, open field activity, sensorimotor reflex test/physiological measurements were conducted. Results: 1. In the home cage observation, there was not observed any abnormal signs in rats. 2. In the observation of open field activity, the reduction of number of unit areas crossed and rearing count was observed caused by Sweet BV treatment. 3. In the observation of handling reactivity, there was not observed any abnormal signs in rats. 4. In the observation of sensorimotor reflex tests/physiological measurements, there was not observed any neurotoxic signs in rats. 5. In the measurement of rectal temperature, treatment of Sweet BV did not showed great influences in the body temperature of rats. Conclusions: Above findings suggest that Sweet BV is relatively safe treatment in the central nervous system. But in the using of over dose, Sweet BV may the cause of local pain and disturbance of movement. Further studies on the subject should be conducted to yield more concrete evidences.

• Molecules and Cells
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• 제23권3호
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• pp.370-378
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• 2007

A superoxide dismutase was purified 62-fold in seven steps to homogeneity from Methylobacillus sp. strain SK1, an obligate methanol-oxidizing bacterium, with a yield of 9.6%. The final specific activity was 4,831 units per milligram protein as determined by an assay based on a 50% decrease in the rate of cytochrome c reduction. The molecular weight of the native enzyme was estimated to be 44,000. Sodium dodecyl sulfate gel electrophoresis revealed two identical subunits of molecular weight 23,100. The isoelectric point of the purified enzyme was found to be 4.4. Maximum activity of the enzyme was measured at pH 8. The enzyme was stable at pH range from 6 to 8 and at high temperature. The enzyme showed an absorption peak at 280 nm with a shoulder at 292 nm. Hydrogen peroxide and sodium azide, but not sodium cyanide, was found to inhibit the purified enzyme. The enzyme activity in cell-free extracts prepared from cells grown in manganese-rich medium, however, was not inhibited by hydrogen peroxide but inhibited by sodium azide. The activity in cell extracts from cells grown in iron-rich medium was found to be highly sensitive to hydrogen peroxide and sodium azide. One mol of native enzyme was found to contain 1.1 g-atom of iron and 0.7 g-atom of manganese. The N-terminal amino acid sequence of the purified enzyme was Ala-Tyr-Thr-Leu-Pro-Pro-Leu-Asn-Tyr-Ala-Tyr. The superoxide dismutase of Methylobacillus sp. strain SK1 was found to have antigenic sites identical to those of Methylobacillus glycogenes enzyme. The enzyme, however, shared no antigenic sites with Mycobacterium sp. strain JC1, Methylovorus sp. strain SS1, Methylobacterium sp. strain SY1, and Methylosinus trichosproium enzymes.

• The Plant Pathology Journal
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• 제38권3호
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• pp.212-219
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• 2022

Fusarium root rot caused by the soil-borne fungus Fusarium solani is one of the most important fungal diseases of cassava in Thailand, resulting in high yield losses of more than 80%. This study aimed to investigate if the exogenous application of salicylic acid formulations (Zacha) can induce resistance in cassava against Fusarium root rot and observe the biochemical changes in induced cassava leaf tissues through synchrotron radiation based on Fourier-transform infrared (SR-FTIR) microspectroscopy. We demonstrated that the application of Zacha11 prototype formulations could induce resistance against Fusarium root rot in cassava. The in vitro experimental results showed that Zacha11 prototype formulations inhibited the growth of F. solani at approximately 34.83%. Furthermore, a significant reduction in the disease severity of Fusarium root rot disease at 60 days after challenge inoculation was observed in cassava plants treated with Zacha11 at a concentration of 500 ppm (9.0%). Population densities of F. solani were determined at 7 days after inoculation. Treatment of the Zacha11 at a concentration of 500 ppm resulted in reduced populations compared with the distilled water control and differences among treatment means at each assay date. Moreover, the SR-FTIR spectral changes of Zacha11-treated epidermal tissues of leaves had higher integral areas of lipids, lignins, and pectins (1,770-1,700/cm), amide I (1,700-1,600/cm), amide II (1,600-1,500/cm), hemicellulose, lignin (1,300-1,200/cm), and cellulose (1,155/cm). Therefore, alteration in defensive carbohydrates, lipids, and proteins contributed to generate barriers against Fusarium invasion in cassava roots, leading to lower the root rot disease severity.

• 식물병연구
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• 제21권3호
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• pp.243-249
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• 2015

점무늬병은 콩에서 최대 50% 정도의 수량감소가 보고된 병으로 최근 한국 콩 재배포장에서 큰 문제로 대두되고 있어 최근 6년간의 보급 품종을 대상으로 콩 점무늬병에 대한 저항성 검정을 실시하였다. 표준화된 검정방법을 적용하기 위해 가장 효과적인 포자 형성 배지 조건을 탐색하고 저항성 검정 평가 기준을 확립하였다. 포자형성은 $25^{\circ}C$, 12시간 광 조건과 암 조건에서 V8 juice를 이용한 배지의 포자 형성이 가장 효과적임을 알 수 있었다. 접종은 $10^5spores/ml$로 준비하여, 제 5 복엽 완전전개기인 V6 stage까지 키운 건강한 콩 식물체에 실시하였고 병 반응은 28일 동안 관찰하였다. 그 결과 대풍은 8개 균주 모두 저항성을 보였고, 신팔달2호는 7개 균주에, 연풍, 청아는 6개 균주에 저항성을 보였다. 반면에 황금, 태광, 대원, 천상, 신화 등은 평가한 8개 균주에 모두 감수성 반응을 나타내었다. 콩 점무늬병에 대한 저항성 품종을 육종하기 위해 표준화되고 정밀화된 저항성 검정방법과 판정 체계를 갖추는 것은 매우 중요하며 이 방법은 차후 포장 저항성 검정에도 적용할 수 있을 것이라 기대한다.