• Title/Summary/Keyword: pre-culture time

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High-efficiency and Rapid Agrobacterium-mediated genetic transformation method using germinating rice seeds (벼 발아초기 종자를 이용한 고효율 단기형질전환 방법)

  • Lee, Hye-Jung;Abdula, Sailila E.;Jee, Moo-Geun;Jang, Dae-Won;Cho, Yong-Gu
    • Journal of Plant Biotechnology
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    • v.38 no.4
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    • pp.251-257
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    • 2011
  • Rice is the most important crop as a model plant for functional genomics of monocotyledons. Rice is usually transformed using Agrobacterium tumefaciens. However, the transformation efficiency using previous method is still low. In this study, we established a new method by modifying the general Agrobacterium protocol especially in the inoculation and co-cultivation step. We directly inoculated Agrobacterium containing a CIPK15 gene under the control of CaMV 35S promoter and NOS terminator in the pCAM1300 vector into the pre-soaked seeds in N6D media for 24 hours. After 7 days of culture at $25^{\circ}C$, calli were formed on seeds cultured on the co-cultivation medium containing an antioxidant compound (1 mM dithiothreitol) and of Agrobacterium growth-inhibiting agent (3 mg/L silver nitrate). We obtained 35 and 22 transgenic plants in rice cultivars, Gopumbyeo and Ilpumbyeo, with increase of transformation efficiency by 30.4% and 22.6%, respectively compared to the general transformation method. The new method in this study would lead to reduction of substantial labor and time to generate transgenic plants.

Effects of Lipomyces starkeyi KSM 22 Glucanhydrolase on human gingival fibroblasts (Lipomyces starkeyi KSM 22 Glucanhydrolase 용액의 치은 섬유아세포에 대한 영향)

  • Yun, Hyun-Jeong;Chung, Hyun-Ju;Kim, Ok-Su;Kim, Do-Man
    • Journal of Periodontal and Implant Science
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    • v.32 no.3
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    • pp.665-683
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    • 2002
  • A novel glucanhydrolase from a mutant of Lipomyces starkeyi KSM 22 has additional amylase activity besides mutanolytic activity and has been suggested as promising anti-plaque agent. It has been shown effective in hydrolysis of mutan, reduction of mutan formation by Streptococcus mutans and removal pre-formed sucrose-dependent adherent microbial film and has been strongly bound to hydroxyapatitie. These in vitro properties of Lipomyces starkeyi KSM 22 glucanhydrolase are desirable for its application as a dental plaque control agent. In human experimental gingivitis model and 6 month clinical trial, mouthrinsing with Lipomyces starkeyi KSM 22 dextranase was comparable to 0.12% chlorhexidine mouthwash in inhibition of plaque accumulation and gingival inflammation and local side effect was negligible. This study was aimed to evaluate the cytotoxic effect of Lipomyces starkeyi KSM 22 glucanhydrolase on human gingival fibroblasts. Primary culture of human gingival fibroblasts at the 4th to 6th passages were used. Glucanhydrolase solution was made from lyophilized glucanhydrolase powder from a mutant of Lipomyces stakeyi KSM 22 solved in PBS and added to DMEM medium to the final concentration of 0.5, 1, and 2 unit. Cells were exposed to glucanhydrolase solution or 0.1 % chlorhexidine and the cells cultured in DMEM with 10% FBS and 1% antibiotics as control. After exposure, the morphological change, cell attachment, and cell activity by MTT assay were evaluated in 0.5, 1.5, 3, 6, 24 hours after treatment. The cell proliferation and cell activity was also evaluated at 2 and 7 days after 1 minute exposure, twice a day. The cell morphology was similar between the Lipomyces smkeyi KSM 22 glucanhydrolase groups and control group during the incubation periods, while most fibroblasts remained as round cell regardless of incubation time in the chlorhexidine group. The numbers of the attached cells in the glucanhydrolase groups were comparable to that of control and significantly higher than the chlorhexidine group. The numbers of the proliferated cells in the glucanhydrolase groups at 7 days of incubation were comparable to the control group and higher than the chlorhexidine group. The cell activity in glucanhydrolase groups paralleled with the increased cell number by attachment and proliferation. According to these results, Lipomyces starkeyj KSM 22 glucanhydrolase has little harmful effect on attachment and proliferation of human gingival fibroblasts, in contrast to 0.1% chlorhexidine which was cytotoxic to human gingival fibroblasts. Therefore this glucanhydrolase preparation is considered as a safe and promising agent for new mouthwash formula in the near future.

LITERATURE REVIEW OF ANTIBIOTICS PRESCRIPTION IN GENERAL DENTAL AND ORAL-MAXILLOFACIAL SURGICAL PRACTICE (치과분야 항생제 처방에 대한 국내외 문헌 분석)

  • Jo, Sae-Hyung;Alrashdan, Mohammad;Choung, Han-Ul;Pang, Kang-Mi;Park, Jong-Chul;Kim, Soung-Min;Kim, Myung-Jin;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.35 no.3
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    • pp.164-169
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    • 2009
  • Objectives : Administration methods of antibiotics implements a variety for indications and diseases. Therefore, it is impossible to produce a single guideline. Many antibiotics have been prescribed without specific index, by practicians for a long period of time. In general dental practice and oral and maxillofacial surgical practice, there is not a guidelines for antibiotics. Therefore, there is a dependency on pre-existent methods rather than following an exact guideline. Consequently, a controversy was issued that prescription of antibiotics tend to be misused or abused. And it is also direct relation to the tolerance of antibiotics as well. Moving forward, in this review we will be analyzing the exact usage and indication of antibiotics in dental treatment. Materials and Methods : 29 published articles of both domestic and international were researched through Pubmed and Kormed, and the review from these articles were performed accordingly. We examined the administration of antibiotics through the objective usage, that classified prophylactic and therapeutic. Results : For therapeutic usage, empirical prescription is mandatory for the first step. Next is to moderate the empirical prescription depending upon the result of its culture. Administration of antibiotics must be kept for 2days after the symptoms disappear. For a prophylactic use, we can generally pr escribe antibiotics to prevent local infection or systemic infection. Although the method of prescription and neccessity of antibiotics to prevent local in fection are controversial, exact guidelines of antibiotics to prevent systemic infection are established by AHA. Conclusion : Most crucial concept for prescript antibiotics is to determine if it is adequately suitable for all circumstances. In this decision making, a guideline for prescription of antibiotics in various dental surgical practice is necessary. This guideline can reduce the misusage and disusage of a ntibiotics in general dental practices and oral and maxillofacial surgical practices.

One step multiplex RT-PCR preventing DNA carryover contamination for differential diagnosis of swine influenza viruses (DNA 교차 오염 방지 기능을 가진 돼지 인플루엔자바이러스 감별진단용 one-step multiplex RT-PCR 진단법)

  • Kim, Hee-Jung;Kim, Eun-Mi;Shin, Yeun-Kyung;Song, Jae-Young;Kim, Seong-Hee;Lee, Kyoung-Ki;Lee, Myoung-Heon;Kim, Young-Hwa;Park, Jun-Cheol;Yeo, Sang-Geon;Park, Choi-Kyu
    • Korean Journal of Veterinary Service
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    • v.37 no.4
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    • pp.263-271
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    • 2014
  • In this study, we developed a cost and time saving one-step multiplex RT-PCR for the simultaneous detection and differentiation of swine influenza viruses (SIV) and 2009 pandemic influenza H1N1 virus (pH1N1). The one-step multiplex RT-PCR using four sets of primer was confirmed to be capable of detection of all SIV subtypes and differential diagnosis of major SIV subtype H1, H3 and pH1N1 on individual or mixed viral culture samples. The sensitivity of the multiplex RT-PCR was determined to be at least $2^{-6}$ $HA/25{\mu}L$ of the presented SIVs, providing sufficient efficacy for a routine SIV monitoring in diagnostic laboratories. In addition, compared with the conventional RT-PCR methods that cannot avoid the carryover DNA contamination, the developed RT-PCR applied with the uracil DNA glycosylase (UNG) system was proven to prevent a false positive reaction by carryover contamination of the pre-amplified DNA. In conclusion, the one-step RT-PCR with UNG system could be applicable to detect and differentiate of SIV from the viral cultures without worry of carryover DNA contamination in clinical laboratories.

Aging and Poverty -Focusing on Age Group Differences in Poverty Trajectories- (노인되기와 빈곤 -빈곤 궤적의 연령집단 차이를 중심으로)

  • Lee, Ji-In;Joo, Eun-Sun
    • Journal of the Korea Convergence Society
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    • v.11 no.5
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    • pp.261-273
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    • 2020
  • The purpose of this study is to examine the trajectories of multi-dimensional poverty in the process of transitioning from middle age to old age, and to identify the factors that influence them. Using the Korea Welfare Panel Data(2006 ~ 2018), we examined the trajectory of changes in multi-dimensional poverty for 13 years by prospective elderly and middle aged group aged 55 or older in 2006 through the potential growth model. Multidimensional poverty is divided into seven dimensions in four areas: economy (income, employment), environment (residential), health, social culture (leisure, family relations, and social relations). The results showed that the level of multi-dimensional poverty decreased with time, and the overall poverty level was higher than that of the pre-old and the average. As a result of analyzing the condition model with the independent variable, the variables affecting poverty change were found in the order of spouse free and educational level for the elderly and gender and education level for the elderly. In other words, multi-dimensional poverty is gradually improving, but the rate of change and the variables that affect each age group are different.

A Literary Study on the Changes in the Meaning of Gongye During Korea's Transition to the Modern Era (근대 전환기 한국 '工藝(공예)' 용어의 쓰임과 의미 변화에 대한 고찰)

  • ROH, Junia
    • Korean Journal of Heritage: History & Science
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    • v.54 no.3
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    • pp.192-203
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    • 2021
  • This paper investigates the process of change in the meaning of the term gongye (工藝) (nowadays it means craft) during Korea's transition to the modern era. The Annals of the Joseon Dynasty (朝鮮王朝實錄) will serve as the basis for the examination of pre-modern usage, and the Korea-United States Treaty of 1882 for the changes that occurred between the translation from Chinese and from Japanese following the opening of the port . After the port opened, the meaning of gongye, which once broadly referred to the wide range of work of people from different fields, was narrowed down to singularly represent the manufacturing industry. Following the example of Japan, the first East Asian country to succeed in modernization, this new meaning of gongye was generally used in the context of embracing Western culture and creating industry. As industrialization emerged as a national mission, and new terminology was needed to express these new concepts, the Chinese characters used by the Japanese to translate Western concepts were directly imported and used without making any changes. Gongye was similar in meaning to gongeop (工業) (nowadays it means industry) at the time, and the two terms were used interchangeably. However, following the Japanese annexation of Korea, the cultural administration of the Governor-General imbued the term with artistic concepts, and gongye was differentiated from gongeop, As such, examining the establishment of terminology and concepts can be helpful for understanding the contradictions and problems of the Korean crafts scene, which were derived from a period in the past called the modern era.

Measurement of Image Quality According to the Time of Computed Radiography System (시간에 따르는 CR장비의 영상의 질평가)

  • Son, Soon-Yong;Choi, Kwan-Woo;Kim, Jung-Min;Jeong, Hoi-Woun;Kwon, Kyung-Tae;Hwang, Sun-Kwang;Lee, Ik-Pyo;Kim, Ki-Won;Jung, Jae-Yong;Lee, Young-Ah;Son, Jin-Hyun;Min, Jung-Whan
    • Journal of radiological science and technology
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    • v.38 no.4
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    • pp.365-374
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    • 2015
  • The regular quality assurance (RQA) of X-ray images is essential for maintaining a high accuracy of diagnosis. This study was to evaluate the modulation transfer function (MTF), the noise power spectrum (NPS), and the detective quantum efficiency (DQE) of a computed radiography (CR) system for various periods of use from 2006 to 2015. We measured the pre-sampling MTF using the edge method and RQA 5 based on commission standard international electro-technical commission (IEC). The spatial frequencies corresponding to the 50% MTF for the CR systems in 2006, 2009, 2012 and 2015 were 1.54, 1.14, 1.12, and $1.38mm^{-1}$, respectively and the10% MTF for 2006, 2009, 2012, and 2015 were 2.68, 2.44, 2.44, and $2.46mm^{-1}$, respectively. In the NPS results, the CR systems showed the best noise distribution in 2006, and with the quality of distributions in the order of 2015, 2009, and 2012. At peak DQE and DQE at $1mm^{-1}$, the CR systems showed the best efficiency in 2006, and showed better efficiency in order of 2015, 2009, and 2012. Because the eraser lamp in the CR systems was replaced, the image quality in 2015 was superior to those in 2009 and 2012. This study can be incorporated into used in clinical QA requiring performance and evaluation of the performance of the CR systems.

Longitudinal Study of the Subgingival Microbial Change after Tetracycline Topical Application (Tetracycline 국소도포가 치은연하 세균분포에 미치는 영향)

  • Choi, Kwang-Choon;Lee, Young-Hee;Lee, Jin-Yong;Chung, Chong-Pyoung;Son, Seong-Heui
    • The Journal of the Korean Society for Microbiology
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    • v.21 no.4
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    • pp.503-513
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    • 1986
  • Previous studies have developed the technique of topical application of tetracycline(TC) into the periodontal pockets and examined the change of clinical parameters and subgingival microbial morphotypes. The purpose of this study was to longitudinally examine the clinical and microbiological effects of topically applied TC in a double-blind and split-mouth design. Thirteen patients with moderate periodontitis, who were treated with or without TC application and scaling treatment, were examined. TC gel(3%) was used to apply into the selected periodontal pockets twice a week for 2 weeks. During the experiment, clinical parameters and subgingival microbial morphotypes were examined, and for isolation of black-pigmented Bacteroides(BPB) and streptococci, an anaerobic sample culturing was done at week 0, 2, and 7. In clinical observation the TC-scaled group exhibited a significant decrease of Gingival Inflammatory Index, Plaque Index, Sulcus Bleeding Index, pocket depth, and gingival crevicular fluid when compared to the TC-unsealed, placebo-scaled, and placebo-unsealed groups. The result of microbial morphotype observation showed a significant increase of coccal form and a decrease of spirochetes in the TC-scaled, TC-unscaled, and placebo-scaled groups. The culture study of streptococci revealed that TC with scaling treatment resulted in a significant increase of S. sanguis I at week 2, but its proportion had returned to the base line level. The anaerobic culture study showed that BPB was significantly reduced in the TC-scaled and TC-unsealed groups at week 7. Among BPB species, B. intermedius declined significantly with time treatment(week 2 and 7) in the TC-scaled and TC-unsealed groups. These results suggest that the settled pathogenic microflora can be succeeded by nonpathogenic microflora in periodontal pockets after TC treatment.

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Human Lung Cancer Cell Xenografts Implanted under the Capsule of Kidney, Spleen and Liver (폐암 세포주를 사용한 신, 비장 및 간 피막하 분식법의 비교)

  • 김수현;김종인;이해영;조봉균;박성달;김송명
    • Journal of Chest Surgery
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    • v.36 no.10
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    • pp.711-720
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    • 2003
  • Bakground : Complete resection by the surgery has been selected as the treatment of choice in lung cancer patients, but in cases of recurrence after excision or inoperable cases, the importance of anticancer chemotherapy has been emphasized. If one can select a set of the sensitive chemotherapeutic agents before anticancer chemotherapy, it will give more favourable results. Subrenal capsular assay has been recognized as a useful in-vivo chemosensitivity test of thoracic and abdominal tumors and it can be done in a short time for a rapid interpretation of tumor responsiveness to anticancer chemotherapeutic drugs. It has been reported that various kinds of cancer cells can be implantable to the kidney, but so far there is no comparative study of xenogeneic cell implantation on liver, spleen and kidney. The author implanted the human lung cancer cells under the capsule of S.D rat's liver, spleen and kidney respectively and compared the pattern of growth and histology. Material and Method: After incubation of human lung cancer cell line (SW-900 G IV) in RPMI 1640 (Leibovitz L-15 medium) culture media, 3${\times}$3${\times}$3 mm size fibrin clots which contain 108 cancer cells were made. Thereafter the fibrin clots were implanted at subcapsule area of liver, spleen and kidney of S.D. female rat. For immune suppression, cyclosporin-A (80 mg/Kg) was injected subcutaneously daily from post-implantation first day to sixth day. The body weight was measured at pre and post implantation periods. The growth pattern and the size of tumor mass were observed and the pathologic examination and serum tumor marker tests were performed. Result: Body weight increased in both of control and experimental groups. Serum Cyfra 21-1 was not detected. Serum levels of CEA and NSE revealed no significant change. The SCC-Ag increased significantly in implanted group. The growth rate of human lung cancer cells which was implanted on spleen was higher than on liver or kidney. The surface area, thickness, and volume of tumor mass were predominant at spleen. The success rates of implantation were 80% on kidney, 76.7% on spleen and 43.3% on liver. Pathologic examination of implanted tumors showed characteristic findings according to different organs. Tumors that were implanted on kidney grew in a round shape, small and regular pattern. In the spleen, tumors grew well and microscopic neovascularization and tumor thrombi were also found, but the growth pattern was irregular representing frequent daughter mass. Human lung cancer cells that were implanted in the liver, invaded to the liver parenchyme, and had low success rate of implantation. Microscopically, coagulation necrosis and myxoid fibrous lesion were observed. Conclusion: The success rate of implantation was highest in the kidney. And the mass revealed regular growth that could be measured easily. The SCC-Ag was presented earlier than CEA or Cyfra21-1. The Cyfra21-1 was not detected at early time after implantation. The best model for tumor implantation experiment for chemosensitivity test was subrenal capsular analysis than liver and spleen and the useful serum tumor marker in early period of implantation was the SCC-Ag.

Experimental Pathogenesis of Pullorum Disease with the Local Isolate of Salmonella enterica serovar. enterica subspecies Pullorum in Pullets in Bangladesh

  • Haider, M.G.;Chowdhury, E.H.;Khan, M.A.H.N.A.;Hossain, M.T.;Rahman, M.S.;Song, H.J.;Hossain, M.M.
    • Korean Journal of Poultry Science
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    • v.35 no.4
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    • pp.341-350
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    • 2009
  • The research work was carried out to study the pathogenesis covering the clinical signs, gross and histopathological lesions in different organs, and reisolation and identification of the organisms after experimental infection with the local isolate of Salmonella enterica serovar. enterica subspecies (S.) Pullorum at different time interval of the experiment during the period February 2006 to December 2006. One hundred pullets (seronegative to S. Pullorum of 12 weeks age were purchased and divided into 5 (A, B, C, D and E) groups and each group consisted of 20 birds. Four groups (A, B, C and D) were infected orally with a dose of $10^6\;CFU$, $10^7\;CFU$, $2{\times}10^7\;CFU$, $10^8\;CFU$ of S. Pullorum, respectively, and one group (E) was treated as uninfected control. The used methods were necropsy and histopathology, culture of bacteria, staining and biochemical test of Salmonella. Five birds from each group were randomly selected and sacrificed $1^{st}$ week, $2^{nd}$, $3^{rd}$ and $4^{th}$ weeks of post infection (PI). From all the groups, the bacteriological samples (crop, liver, lung, heart, spleen, bile duodenum, ceca and blood) were collected with pre enriched in buffered peptone water in sterile poly bags. Liver, lungs, heart, spleen, intestine, etc. were collected in 10% buffered-formalin for histopathological examination. No clinical signs, gross and histopathological lesions were found in control group and no S. Pullorum was reisolated. Clinical sign of experimentally infected with S. Pullorum in pullets were loss of appetite (100%), slight depression (75%), ruffled feathers (85%), diarrhea (60%) and loss of weight (100%) in chickens. The feed intake and body weight at different weeks after PI differed significantly (p<0.01) among the groups. Grossly, the highest recorded lesion was button-like ulcer in the ceca (80%) and the lowest was white nodules in lungs (1.25%). S. Pullorum were reisolated from crop (91.25%), liver (91.25%), lung (83.75%), heart (71.25%), spleen (87.75%), bile (33.25%), duodenum (92.50%), ceca (97.50%) and from different group of infection (61.25%). The highest microscopic findings were intestinal and cecal mucosa and submucosa exhibited infiltration of mononuclear cells and congestion (96.25%), and the lowest finding was nodule formation in the lungs (3.75%). The pattern of the disease production by local isolate of S. Pullorum in Bangladesh is almost similar with other isolates in different countries.