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Overproduction of Sodium Gluconate Using the Recombinant Aspergillus niger (재조합 Aspergillus niger에 의한 글루콘산나트륨의 산업적 생산)

  • 이선희;이현철;김대혁;양문식;정봉우
    • KSBB Journal
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    • v.13 no.2
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    • pp.214-219
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    • 1998
  • Polymerase chain reaction(PCR) was conducted to obtain the gene encoding glucose oxidase(GOD) from Aspergillus niger(ATCC 2110) and the DNA sequence determined was coincided with published GOD sequence from A. niger. Recombinant transforming vector containing GOD and hygromycin B(hyg.B) resistant gene(hph) was constructed and used for further transformation of A. niger ATCC 2110. Selectivity of hyg.B against A. niger differed depending on which media were used i.e., nutrient-rich media such as potato dextrose agar(PDA) and complete medium(CM) showed only 50% growth inhibition at 400 $\mu$m ml$^-1$ of hyg.B while the minimal media inhibited mycelial growth completely at 200 $\mu$m ml$^-1$ of hyg.B. Twenty to sixty putative transformants were isolated from the hyg.B-containing minimal top agar, transferred successively onto alternating selective and nonselective media for a mitotic stability of hyg.B resistance and, then, single-spored. Among the stable transformants, the transformant(GOD1-6) grown by flask culture showed the considerable increase of extracellular GOD activity, which was estimated to the degree of 50% - 100% comparing to that of wild type. Transformation of tGOD1-6 was resulted from integration of the vectors into heterologous as well as homologous regions of the A. niger genome. Southern blot analysis revealed that there were two independent integrations of vector into fungal genome and one into the GOD gene due to homologous recombination. In addition, GOD activity and sodium gluconate production when tGOD1-6 was fed-batch fermented were enhanced 11 fold and 2.25 fold, respectively, compared to that of the wild type.

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Chondroprotective and Anti-inflammatory Effects of ChondroT, A New Complex Herbal Medication

  • Jung Up Park;WonWoo Lee
    • Proceedings of the Plant Resources Society of Korea Conference
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    • 2022.09a
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    • pp.103-103
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    • 2022
  • Ganghwaljetongyeum (GHJTY) is a complex herbal decoction comprising 18 plants; it is used to treat arthritis. In order to develop a new anti-arthritic herbal medication, we selected 5 out of 18 GHJTY plants by using bioinformatics analysis. The new medication, called ChondroT, comprised water extracts of Osterici Radix, Lonicerae Folium, Angelicae Gigantis Radix, Clematidis Radix, and Phellodendri Cortex. This study was designed to investigate its chondroprotective and anti-inflammatory effects to develop an anti-arthritic herb medicine. ChondroT was validated using a convenient and accurate high-performance liquid chromatography. photodiode array (HPLC-PDA) detection method for simultaneous determination of its seven reference components. The concentrations of the seven marker constituents were in the range of 0.81-5.46 mg/g. The chondroprotective effects were evaluated based on SW1353 chondrocytes and matrix metalloproteinase 1 (MMP1) expression. In addition, the anti-inflammatory effects of ChondroT were studied by Western blotting of pro-inflammatory enzymes and by enzyme-linked immunosorbent assay (ELISA) of inflammatory mediators in lipopolysaccharides (LPS)-induced RAW264.7 cells. ChondroT enhanced the growth of SW1353 chondrocytes and also significantly inhibited IL-1β-induced MMP-1 expression. However, ChondroT did not show any effects on the growth of HeLa and RAW264.7 cells. The expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) was induced by LPS in RAW264.7 cells, which was significantly decreased by pre-treatment with ChondroT. In addition, ChondroT reduced the activation of NF-κB and production of inflammatory mediators, such as IL-1β, IL-6, PGE2, and nitric oxide (NO) in LPS-induced RAW264.7 cells. These results show that ChondroT exerted a chondroprotective effect and demonstrated multi-target mechanisms related to inflammation and arthritis. In addition, the suppressive effect was greater than that exhibited by GHJTY, suggesting that ChondroT, a new complex herbal medication, has therapeutic potential for the treatment of arthritis.

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Design and Implementation of Unified Index for Moving Objects Databases (이동체 데이타베이스를 위한 통합 색인의 설계 및 구현)

  • Park Jae-Kwan;An Kyung-Hwan;Jung Ji-Won;Hong Bong-Hee
    • Journal of KIISE:Databases
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    • v.33 no.3
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    • pp.271-281
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    • 2006
  • Recently the need for Location-Based Service (LBS) has increased due to the development and widespread use of the mobile devices (e.g., PDAs, cellular phones, labtop computers, GPS, and RFID etc). The core technology of LBS is a moving-objects database that stores and manages the positions of moving objects. To search for information quickly, the database needs to contain an index that supports both real-time position tracking and management of large numbers of updates. As a result, the index requires a structure operating in the main memory for real-time processing and requires a technique to migrate part of the index from the main memory to disk storage (or from disk storage to the main memory) to manage large volumes of data. To satisfy these requirements, this paper suggests a unified index scheme unifying the main memory and the disk as well as migration policies for migrating part of the index from the memory to the disk during a restriction in memory space. Migration policy determines a group of nodes, called the migration subtree, and migrates the group as a unit to reduce disk I/O. This method takes advantage of bulk operations and dynamic clustering. The unified index is created by applying various migration policies. This paper measures and compares the performance of the migration policies using experimental evaluation.

Clinical Analysis Of Ventricular Septal Defect (심실중격결손증의 외과적 고찰)

  • Seong, Suk-Hwan;Suh, Kyung-Pill
    • Journal of Chest Surgery
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    • v.15 no.1
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    • pp.90-97
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    • 1982
  • Two hundred one patients of ventricular septal defect, which were operated at Seoul National University Hospital, were analysed on clinical background during the period from January, 1975 to December, 1980. The results were as follows: 1. Of the 201 patients, 118 patients were male [58.7%] and 83 patients were female [41.3%]. Their age ranged from 15 months to 40 years, and the mean age was 8.7 years. 40% of the patients were between 4 and 8 years. 2. The most common symptoms showed frequent U RI and exertional dyspnea. 3. On Kirklin`s anatomical classification, type I constituted 26.9%, type II 58.2%, type III 12.4%, and type IV 1.0%. We showed marked increased incidence of type I VSD as compared to Caucasians* 4. 46 cases were associated with other congenital cardiac diseases. They were PDA [13 cases], AI[11 ], ASD[6], PS[10], MI[4], and Double aortic arch [1]. 5. In 128 patients, who had complete hemodynamic data and were not associated with other congenital cardiac diseases, an attempt was made to correlate the EKG findings with the hemodynamic data, and defect size with the hemodynamic data. The children had variable distribution of PA syst. pr. and Rp/Rs. But most of adults had $R_P$/$R_S$of 0.15 or less. As $P_P$/$P_S$increased, the rate of operative complication increased also. 6. When a normal EKG pattern was present, $Q_P$/$Q_S$and $R_P$/$R_S$and $P_P$/$P_S$were relatively low. When EKG findings were LVH pattern, there was diastolic volume overload to left ventricle. As RVH, there was systolic pressure overload to right ventricle. And as BVH, there was mixed pattern of diastolic volume overload to left ventricle and systolic pressure overload to right ventricle. 7. Among patients in defect was less than 1 $cm^2$ per $M^2$ of BSA, $Q_P$/$Q_S$was less than 2:1, and $R_P$/$R_S$less than 0.25, and PAsyst. pr. less than 50 mmHg, and $P_P$/$P_S$was less than 0.5. But patients with the defect greater than 1 $cm^2$ per $M^2$ of BSA had no correlationship between $Q_P$/$Q_S$, $R_P$/$R_S$, PAsyst. pr. and defect size in each other. Most of patients with the defect greater than 2 $cm^2$/$M^2$ BSA, $R_P$/$R_S$was greater than 0.5. 8. Operative mortality rate was 9.5% [19 cases] among 201 patients. And complication rate including mortality rate was 22.9% [46 cases].

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Characteristics of Fluoride-based Anti-stain Chemicals Made from industrial By-product (I) - Anti-mold Effectiveness, Iron Corrosivity and Hygroscopicity - (산업 부산물을 이용하여 제조한 플루오르화합물계 목재 방미제의 특성 (I) - 목재 방미효력 및 철부식성과 흡습성 -)

  • Lee, Jong-Shin
    • Journal of the Korean Wood Science and Technology
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    • v.32 no.2
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    • pp.73-81
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    • 2004
  • To develope of the inexpensive anti-stain chemicals, it was conducted to investigate the inhibitory effectiveness of mycelial growth and anti-mold effectiveness against test fungi causing surface mold of wood, iron corrosivity and hygroscopicity of six fluoride-based chemicals made from industrial by- product.Among the six chemicals. RNF-3 and RNF-4 were the most effective with respect to a mycelial growth control and anti-mold effectiveness. For the wood treatment with these chemicals, the optimum concentration was about 2% and there was no difference in the effectiveness of chemicals between wood species. The wood treated with synthesized chemicals showed a relatively high iron corrosion rate for corrosive F component whereas, in the treatment with RNF-3 and RNF-4, there was no difference from the untreated wood when the concentration was less than 2%. The hygroscopic property of wood was not effected by treatment of these chemicals. To prove the feasibility for practical using of RNF-3 and RNF-4 chemicals, it is necessary to test of anti-mold effectiveness in the sawmill by field test.

Comparison of the bioactive compounds and anti-inflammatory effects found in different flower colors from Abeliophyllum distichum Nakai (미선나무 꽃 색에 따른 생리활성 화합물 및 항염증 활성 비교)

  • Jang, Tae-Won;Choi, Ji-Soo;Han, So-Yeon;Park, Hye-Jeong;Lee, Da-Yoon;Min, Young-Sil;Park, Jae-Ho
    • Journal of Applied Biological Chemistry
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    • v.65 no.3
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    • pp.203-213
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    • 2022
  • Abeliophyllum distichum (A. distichum, Korean endemic plant) is one genus and one species in the Oleaceae family. According to the color variation of petals and calyx, A. distichum is classified as A. distichum (white flower), A. distichum for. lilacinum (pink flowers), A. distichum for. eburneum (ivory flowers), and Okhwang 1 (golden flowers). In previous studies, bioactivities (antioxidant, anti-inflammatory, and anti-cancer) of A. distichum have been reported. We conducted a comparison of the differences in bioactive compounds and the anti-inflammatory effects on macrophages among four flowers of A. distichum (FAD). The identification and quantification of glycosides were analyzed by HPLC/PDA and LCMS. These results were shown FAD has rutin, hirsutrin, and acteoside. Antioxidant activity of FAD significantly decreased reactive oxygen species. In addition, FAD reduced the expression of pro-inflammatory mediators (nitric oxide, iNOS, and COX-2) in lipopolysaccharide-induced RAW 264.7 cells. For further study, we investigated the regulation of nuclear factor kappa B (NF-κB) and mitogen-activated protein kinase (MAPK) signaling pathways. In conclusion, FAD may exert anti-inflammatory effects by suppressing inflammatory mediators via regulations of NF-κB and MAPK signaling pathways. Therefore, these findings suggest that FAD is a potential resource as a preventative or therapeutic agent for inflammation.

Mass Screening of Lovastatin High-yielding Mutants through Statistical Optimization of Sporulation Medium and Application of Miniaturized Fungal Cell Cultures (Lovastatin 고생산성 변이주의 신속 선별을 위해 통계적 방법을 적용한 Sporulation 배지 개발 및 Miniature 배양 방법 개발)

  • Ahn, Hyun-Jung;Jeong, Yong-Seob;Kim, Pyeung-Hyeun;Chun, Gie-Taek
    • KSBB Journal
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    • v.22 no.5
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    • pp.297-304
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    • 2007
  • For large and rapid screening of high-yielding mutants of lovastatin produced by filamentous fungal cells of Aspergillus terreus, one of the most important stage is to test as large amounts of mutated strains as possible. For this purpose, we intended to develop a miniaturized cultivation method using $7m{\ell}$ culture tube instead of traditional $250m{\ell}$ flask (working volume $50m{\ell}$). For obtaining large amounts of conidiospores to be used as inoculums for miniaturized cultures, 4 components i.e., glucose, sucrose, yeast extract and $KH_2PO_4$ were intensively investigated, which had been observed to show positive effect on enhancement of spore production through Plackett-Burman design experimet. When optimum concentrations of these components that were determined through application of response surface method (RSM) based on central composite design (CCD) were used, maximum spore numbers amounting to $1.9\times10^{10}$ spores/plate were obtained, resulting in approximately 190 fold increase as compared to the commonly used PDA sporulation medium. Using the miniaturized cultures, intensive strain development programs were carried out for screening of lovastatin high-yielding as well as highly reproducible mutants. It was observed that, for maximum production of lovastatin, the producers should be activated through 'PaB' adaptation process during the early solid culture stage. In addition, they should be proliferated in condensed filamentous forms in miniaturized growth cultures, so that optimum amounts of highly active cells could be transferred to the production culture-tube as reproducible inoculums. Under these highly controlled fermentation conditions, compact-pelleted morphology of optimum size (less than 1 mm in diameter) was successfully induced in the miniaturized production cultures, which proved essential for maximal utilization of the producers' physiology leading to significantly enhanced production of lovastatin. As a result of continuous screening in the miniaturized cultures, lovastatin production levels of the 81% of the daughter cells derived from the high-yielding producers turned out to be in the range of 80%$\sim$120% of the lovastatin production level of the parallel flask cultures. These results demonstrate that the miniaturized cultivation method developed in this study is efficient high throughput system for large and rapid screening of highly stable and productive strains.

Optimization of Mycelial Growth of Entomogenous fungi of the Genus Cordyceps (동충하초속균의 균사생장최적화)

  • Hong, In-Pyo;Nam, Sung-Hee;Jung, I-Yeon;Sung, Gyoo-Byung;Nam, Hack-Woo;Kang, Seok-Woo;Hur, Hyeon;Lee, Min-Woong;Guo, Shun-Xing
    • Journal of Mushroom
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    • v.2 no.3
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    • pp.149-156
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    • 2004
  • This study was carried out to obtain basic data on physiological characteristics for an artificial cultivation of fruiting body of Cordyceps. Specimens such as Cordyceps longissima, C. militaris and C. pruinosa were collected at Mt. Halla of Cheju island in July, 2003. Among four different culture media which have been used for culture of mushrooms, MCM medium was selected for the favorable culture medium of the Cordyceps tested. The initial pH of solid medium for mycelial growth of Cordyceps was good in the range of pH 5.0~7.0 lower than 8.0. The mycelial growth of C. longissima was most favorable on culture media supplemented with glucose, one of monosaccharides. In C. militaris, nine carbon sources were favorable to the mycelial growth as compared with control among 11 carbon sources. Six nitrogen sources were favorable to the mycelial growth of C. longissima as compared with control among 9 carbon sources; namely, the mycelial growth of C. longissima was most favorable on culture media contained potassium nitrate, and followed in order by ammonium citrate and sodium nitrate in 4 weeks incubation.

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Surgical outcome of severe pulmonary arterial hypertension secondary to left-to-right shunt lesions (심한 폐동맥 고혈압을 동반한 좌우 단락 질환 환자의 수술 후 경과)

  • Lee, Cha Gon;Jeong, Su-In;Huh, June;Kang, I-Seok;Lee, Heung Jae;Yang, Ji-Hyuk;Jun, Tae Gook
    • Clinical and Experimental Pediatrics
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    • v.53 no.2
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    • pp.195-202
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    • 2010
  • Purpose : Despite recent advances in pulmonary hypertension management and surgery, appropriate guidelines remain to be developed for operability in congenital heart disease with pulmonary artery hypertension (PAH). Our aim was to evaluate clinical outcomes of patients with severe PAH who underwent surgical closure of left-to-right shunt lesions (LRSL) on the basis of pulmonary reactivity. Methods : We retrospectively reviewed 21 patients who underwent surgical closure of LRSL with severe PAH (${\geq}8$ Wood unit) from January 1995 to April 2009. The median age at operation was 26 years. Atrial septal defect, ventricular septal defect (VSD), VSD and patent ductus arteriosus (PDA), and PDA was present in 11, 4, 4, and 2 patients, respectively. Results : Operability was based on vasoreactivity of PAH. Of the 21 patients, 5 showed response to pulmonary vasodilator therapy and 8 showed vasoreactivity after balloon occlusion of defects. The remaining 8 patients were considered operable because of significant left-to-right shunt (Qp/Qs ${\geq}1.5$). Five patients underwent total closure of defects and 16 were left with small residual shunts. The median follow-up duration was 32 months. There was no significant postoperative mortality or morbidity. Systolic pulmonary artery pressure (PAP) decreased in all but 2 patients. All patients except 1 showed improvement of New York Heart Association functional class. Conclusion : Closure of LRSL in patients with severe PAH on the basis of pulmonary vasoreactivity seems reasonable. PAP and clinical symptoms improved in most patients. Further research is needed for the evaluation of long-term results.

Incidence and Control of Bottom Rot of Chinese Cabbage Caused by Rhizoctonia solani Kuhn (배추 밑동썩음병 발생과 방제)

  • Kang Soo Woong;Kim Hee Kyu
    • Korean Journal Plant Pathology
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    • v.2 no.3
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    • pp.193-198
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    • 1986
  • Bottom rot of Chinese cabbage (Brassica pekinensis Rupr) hitherto undescribed in Korea is caused by Rhizoctonia solani Kuhn. Development of bottom rot in fall crops was observed and the efficiencies of fungiciial control in naturally infested field in Southern region were studied. Bottom rot symptom started to develop 20 days after transplanting 20 day old seedlings, which corresponded to the middle growing stage. At the reading stage, the infection rate increased substantially, which progressed upto an average of $52\%$ at harvesting time. The overall infection rate was about $30\%$ in Chinese cabbage field under plastic film house cultivation. However, disease intensity of fall crops was less severe than that of crops in plastic film house. where plants infested earlier, stunted and their growth was extremely poor. Optimum temperature for mycelial growth of bottom rot isolates of R. solani was $20\~25^{\circ}C$ on potato-sucrose agar. This fungus was highly pathogenic on seedlings of Chinese cabbage, radish, sesame and rape resulting in high percentage of damping-off. For other crops, such as lettuce, tomato the cucumber, the germination was delayed for 2-3 days and the percentage damping-off was lower. Anastomosis group of this fungus was idenlfied as AG II-I. Soil drenching of fungicide pencycr WP., three applications at 10 day interval, was effective; indicating the most promising one with control value $80\%$.

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