• Title/Summary/Keyword: Cell selection

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Scopolamine Production in Suspension Cultures of Tumor Calli from Datura metel L. (흰독말풀(Datura metel L.)종양 캘러스의 현탁배양으로부터 Scopolamine 생성)

  • 이수경;윤길영;김용해;양덕조
    • Korean Journal of Plant Tissue Culture
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    • v.27 no.3
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    • pp.203-211
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    • 2000
  • In this study to produce large-scale scopolamine we were examined in the tumor calli of Datura metel L. induced by Agrobacterium tumefaciens $Ery{101}$. The growth and scopolamine contents of tumor calli were higher under light condition than in dark. The optimum condition of growth and scopolamine production were fluence rate of 16 $\mu$mol $m^{-2}s^{-1}$, spectra of red light region and 16 hour light periods on 50 mL SH liquid medium in 4 weeks culture. To increase of the scopolamine contents in tumor calli, the optimum concentration of nitrogen source were 1.8 mM NH$_4$+ and 40 mM NO$_3$. The optimum elicitor concentration for production of scopolamine were 10 mg/L chitosan and 15 mg/L yeast extract. The effect of precursors were good at the concentration of 0.2 mM tropine and 0.3 mM tropic acid, respectively. In order to increase of growth and scopolamine contents. we induced mutant from Datura metel L. tumor callus. Mutants of tumor calli were obtained by 3 Krad, 4 Krad and 6 Krad of ${60}^Cor-ray$. Among them, 3 Krad tumor callus was excellent on the growth and teratoma induction. The 4 Krad tumor callus was negligible for both growth and teratoma induction. But the 6 Krad tumor callus was the best in growth and teratoma induction. The formation of the mutant calli can be enhanced through hormonal combination of 1 mg/L 2,4-dichlorophenoxyacetic acid and 0.5 mg/L benzyladenine. We carry out selection mutant tumor calli for high content tropane alkaloid and suspension cultures for scopolamine production.

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Microsurgical DREZotomy for Deafferentation Pain (구심로 차단 동통에서의 미세 후근 진입부 절제술)

  • Kim, Seong-Rim;Lee, Kyung Jin;Cho, Jeong Gi;Rha, Hyung Kyun;Park, Hae Kwan;Kang, Joon Ki;Choi, Chang Rak
    • Journal of Korean Neurosurgical Society
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    • v.30 no.sup1
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    • pp.85-90
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    • 2001
  • Objective : DREZotomy is effective for the treatment of deafferentation pain as a consequence of root avulsion, postparaplegic pain, posttraumatic syrinx, postherpetic neuralgia, spinal cord injury, and peripheral nerve injury. We performed microsurgical DREZotomy to the patients with deafferentation pain and relieved pain without any serious complication. The purpose of this study is to evaluate the usefulness of the microsurgical DREZotomy for deafferentation pain. Methods : We evaluated 4 patients with deafferntation pain who were intractable to medical therapy. Two of them were brachial plexus injury with root avulsion owing to trauma, one was axillary metastasis of the squamous cell carcinoma of the left forearm, and the last was anesthesia dolorosa after surgical treatment(MVD and rhizotomy) of trigeminal neuralgia. Preoperative evaluation was based on the neurologic examination, radiologic imaging, and electrophysiological study. In the case of anesthesia dolorosa, we produced two parallel lesions in cephalocaudal direction, 2mm in distance, from the C2 dorsal rootlet to the 5mm superior to the obex including nucleus caudalis, after suboccipital craniectomy and C1-2 laminectomy, with use of microelectrode. In the others, we confirmed lesion site with identification of the nerve root after hemilaminectomy. We performed arachnoid dissection along the posterolateral sulcus and made lesion with microsurgical knife and microelectrocoagulation, 2mm in depth, 2mm in distance, to the direction of 30-45 degrees in the medial portion of the Lissauer's tract and the most dorsal layers of the posterior horn at the one root level above and below the lesion. Results : Compared with preoperative state, microsurgical DREZotomy significantly diminished dosage of the drugs and relieved pain meaningfully. One patient showed tansient ipsilateral ataxia, but recovered soon. There was not any serious complication. Conclusion : It may be concluded that microsurgical DREZotomy is very useful and safe therapeutic modality for deafferentation pain, especially segmentally distributed intermittent or evoke pain. Complete preoperative evaluation and proper selection of the patients and lesion making device are needed to improve the result.

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Efficient Construction Method of Topographic Data for Flood Mapping Using Digital Map (수치지형도를 활용한 홍수지도 제작용 지형자료의 효과적인 구축방법 연구)

  • Lee, Geun-Sang;Koh, Deuk-Koo;Kim, Woo-Gu
    • Journal of the Korean Association of Geographic Information Studies
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    • v.7 no.1
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    • pp.52-61
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    • 2004
  • Korea Water Resources Corporation carried out LiDAR survey to construct detailed terrain data for flood mapping and it is expected that much money is required in flood mapping of all over the country. Therefore, it is desirable to use NGIS digital map to construct preliminary modelling data for selection of flood mapping area. And the analysis of DEM error with respect to scale of digital map is necessary for the sake of applying digital map as the input data of flood mapping. We compared DEM from digital map with DEM from LiDAR survey. Especially we analyzed DEM error characteristics that is occurred with respect to the interpolation method that is used to construct DEM from TIN of digital map. As a result of analysis, digital map(1:1,000) showed smaller error than digital map(1:5,000) and DEM applying linear interpolation showed smaller error than DEM applying quintic interpolation. Especially, variation of DEM error by cell resolution was evaluated as very slight because urban district was composed of gentle slope.

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Adjuvant Chemotherapy in Patients with Node-Negative Non-Small Cell Lung Cancer with Satellite Pulmonary Nodules in the Same Lobe

  • Park, Jiyoun;Lee, Junghee;Jeon, Yeong Jeong;Shin, Sumin;Cho, Jong Ho;Kim, Hong-Kwan;Choi, Yong Soo;Kim, Jhingook;Zo, Jae Ill;Shim, Young Mog
    • Journal of Chest Surgery
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    • v.55 no.1
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    • pp.10-19
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    • 2022
  • Background: According to the eighth TNM (tumor-node-metastasis) staging system, the presence of separate tumor nodules in the same lobe is designated as a T3 descriptor. However, it remains unclear whether adjuvant chemotherapy confers survival advantages in this setting. Methods: We retrospectively identified 142 pathologic T3N0M0 patients with additional pulmonary nodules in the same lobe from a single-institutional database from 2004 to 2019. The main outcomes were overall survival and recurrence-free survival. Multivariable Cox regression was used to identify the benefit of adjuvant chemotherapy while adjusting for other variables. Results: Sixty-one patients received adjuvant chemotherapy (adjuvant group) and 81 patients did not receive adjuvant therapy after surgery (surgery-only group). There were no demonstrable differences between the 2 groups regarding hospital mortality and postoperative complications, indicating that treatment selection had not significantly occurred. However, the use of adjuvant chemotherapy was associated with improved 5-year overall survival (70% vs. 59%, p=0.006) and disease-free survival (60% vs. 46%, p=0.040). A multivariable Cox model demonstrated that adjuvant chemotherapy was associated with a survival advantage (adjusted hazard ratio, 0.54; p<0.001). In exploratory analyses of subgroups, the effect of adjuvant chemotherapy seemed to be insufficient in those with small main tumors (<4 cm). Conclusion: Adjuvant chemotherapy was associated with better survival in T3 cancers with an additional tumor nodule in the same lobe. However, the role of adjuvant chemotherapy in patient subgroups with small tumors or those without risk factors should be determined via large studies.

Optimization of Protoplast Isolation and Ribonucleoprotein/Nanoparticle Complex Formation in Lentinula edodes (표고버섯의 원형질체 분리 최적화와 RNPs/나노파티클 복합체 형성)

  • Kim, Minseek;Ryu, Hojin;Oh, Min Ji;Im, Ji-Hoon;Lee, Jong-Won;Oh, Youn-Lee
    • Journal of Mushroom
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    • v.20 no.3
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    • pp.178-182
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    • 2022
  • Despite the long history of mushroom use, studies examining the genetic function of mushrooms and the development of new varieties via bio-molecular methods are significantly lacking compared to those examining other organisms. However, owing to recent developments, attempts have been made to use a novel gene-editing technique involving CRISPR/Cas9 technology and genetic scissors in mushroom studies. In particular, research is actively being conducted to utilize ribonucleoprotein particles (RNPs) that can be genetically edited with high efficiency without foreign gene insertion for ease of selection. However, RNPs are too large for Cas9 protein to pass through the cell membrane of the protoplasmic reticulum. Furthermore, guide RNA is unstable and can be easily decomposed, which remarkably affects gene editing efficiency. In this study, nanoparticles were used to mitigate the shortcomings of RNP-based gene editing techniques and to obtain transformants stably. We used Lentinula edodes (shiitake mushroom) Sanjo705-13 monokaryon strain, which has been successfully used in previous genome editing experiments. To identify a suitable osmotic buffer for the isolation of protoplast, 0.6 M and 1.2 M sucrose, mannitol, sorbitol, and KCl were treated, respectively. In addition, with various nanoparticle-forming materials, experiments were conducted to confirm genome editing efficiency via the formation of nanoparticles with calcium phosphate (CaP), which can be bound to Cas9 protein without any additional amino acid modification. RNPs/NP complex was successfully formed and protected nuclease activity with nucleotide sequence specificity.

Analysis of Infiltration Route using Optimal Path Finding Methods and Geospatial Information (지형공간정보 및 최적탐색기법을 이용한 최적침투경로 분석)

  • Bang, Soo Nam;Heo, Joon;Sohn, Hong Gyoo;Lee, Yong Woong
    • KSCE Journal of Civil and Environmental Engineering Research
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    • v.26 no.1D
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    • pp.195-202
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    • 2006
  • The infiltration route analysis is a military application using geospatial information technology. The result of the analysis would present vulnerable routes for potential enemy infiltration. In order to find the susceptible routes, optimal path search algorithms (Dijkstra's and $A^*$) were used to minimize the cost function, summation of detection probability. The cost function was produced by capability of TOD (Thermal Observation Device), results of viewshed analysis using DEM (Digital Elevation Model) and two related geospatial information coverages (obstacle and vegetation) extracted from VITD (Vector product Interim Terrain Data). With respect to 50m by 50m cells, the individual cost was computed and recorded, and then the optimal infiltration routes was found while minimizing summation of the costs on the routes. The proposed algorithm was experimented in Daejeon region in South Korea. The test results show that Dijkstra's and $A^*$ algorithms do not present significant differences, but A* algorithm shows a better efficiency. This application can be used for both infiltration and surveillance. Using simulation of moving TOD, the most vulnerable routes can be detected for infiltration purpose. On the other hands, it can be inversely used for selection of the best locations of TOD. This is an example of powerful geospatial solution for military application.

Conserved COG Pathways and Genes of 122 Species of Archaea (고세균 122종의 보존적 COG pathways와 유전자)

  • Dong-Geun Lee ;Sang-Hyeon Lee
    • Journal of Life Science
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    • v.33 no.11
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    • pp.944-949
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    • 2023
  • The purpose of this study was to identify conserved metabolic pathways and conserved genes in 122 archaeal species. Using the Clusters of Orthologous Groups of Proteins (COG) database of conserved genes, we analyzed whether 122 species had 63 COG metabolic pathways, the 822 COGs that compose them, and a total of 4,877 COGs. Archaeal ribosomal proteins were the most conserved in metabolic pathways. 46 COGs in seven COG pathways among 63 COG pathways and 20 COGs in others were conserved in 122 species. Some genes involved in cell wall and extracellular matrix synthesis, replication, transcription, translation, and protein metabolism were common to all 122 species. When the distance value of the phylogenetic tree was analyzed at the phylum level or class level, the average was the lowest at the class Halobacteria of the phylum Euryarchaeota. Standard deviation was high for the class Nitosospharia of the phylum Thaumarchaeota, the unclassified members of phylum Thaumarchaeota, the class Halobacteria of the phylum Euryarchaeota, the class Thermoprotei of the phylum Crenarchaeota, and other archaea. Furthermore, the phylogenetic tree analysis revealed six commonalities. The results of this study, along with data on conserved genes, could be used for drug development and gene selection for strain improvement.

Genome-Wide Analysis of DNA Methylation before- and after Exercise in the Thoroughbred Horse with MeDIP-Seq

  • Gim, Jeong-An;Hong, Chang Pyo;Kim, Dae-Soo;Moon, Jae-Woo;Choi, Yuri;Eo, Jungwoo;Kwon, Yun-Jeong;Lee, Ja-Rang;Jung, Yi-Deun;Bae, Jin-Han;Choi, Bong-Hwan;Ko, Junsu;Song, Sanghoon;Ahn, Kung;Ha, Hong-Seok;Yang, Young Mok;Lee, Hak-Kyo;Park, Kyung-Do;Do, Kyoung-Tag;Han, Kyudong;Yi, Joo Mi;Cha, Hee-Jae;Ayarpadikannan, Selvam;Cho, Byung-Wook;Bhak, Jong;Kim, Heui-Soo
    • Molecules and Cells
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    • v.38 no.3
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    • pp.210-220
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    • 2015
  • Athletic performance is an important criteria used for the selection of superior horses. However, little is known about exercise-related epigenetic processes in the horse. DNA methylation is a key mechanism for regulating gene expression in response to environmental changes. We carried out comparative genomic analysis of genome-wide DNA methylation profiles in the blood samples of two different thoroughbred horses before and after exercise by methylated-DNA immunoprecipitation sequencing (MeDIP-Seq). Differentially methylated regions (DMRs) in the pre-and post-exercise blood samples of superior and inferior horses were identified. Exercise altered the methylation patterns. After 30 min of exercise, 596 genes were hypomethy-lated and 715 genes were hypermethylated in the superior horse, whereas in the inferior horse, 868 genes were hypomethylated and 794 genes were hypermethylated. These genes were analyzed based on gene ontology (GO) annotations and the exercise-related pathway patterns in the two horses were compared. After exercise, gene regions related to cell division and adhesion were hypermethylated in the superior horse, whereas regions related to cell signaling and transport were hypermethylated in the inferior horse. Analysis of the distribution of methylated CpG islands confirmed the hypomethylation in the gene-body methylation regions after exercise. The methylation patterns of transposable elements also changed after exercise. Long interspersed nuclear elements (LINEs) showed abundance of DMRs. Collectively, our results serve as a basis to study exercise-based reprogramming of epigenetic traits.

MDM2, p53 and pRb Expression Prior to Definitive Chemoradiotherapy in Esophageal Carcinoma (식도암에서 MDM2, p53, pRb 발현과 동시적 항암화학방사선요법의 결과)

  • Yoon, Mee-Sun;Lee, Jae-Hyuk;Cho, Sang-Hee;Song, Ju-Young;Ahn, Sung-Ja;Chung, Ik-Joo;Chung, Woong-Ki;Nah, Byung-Sik;Nam, Taek-Keun
    • Radiation Oncology Journal
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    • v.25 no.4
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    • pp.193-200
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    • 2007
  • Purpose: This study evaluated the pretreatment expression patterns of MDM2, p53, and pRb proteins to determine if the expression patterns could predict the outcome of concurrent chemoradiotherapy (CCRT) for esophageal squamous cell carcinoma and aid in the decisions for the selection of treatment modalities. Materials and Methods: Fifty-one patients that were treated with definitive chemoradiotherapy for stage $I{\sim}IVa$ esophageal squamous cell carcinoma were selected for this study. Radiotherapy was administered with daily $1.8{\sim}2\;Gy$ fractions up to a median dose of 54 Gy for primary tumors, and with four cycles of cisplatin/5-fluorouracil chemotherapy that was administered every 4 weeks, the first two cycles of which were administered concurrently with radiotherapy. Expression of MDM2, p53, and pRb was investigated by immunohistochemical analysis using pretreatment biopsy specimens. Results: MDM2, p53, and pRb were detected with high immunoreactivity in 19.6%, 27.5%, and 66.7% of the patients, respectively. However, there was no significant correlation between expression of these factors and clinical outcome. By the use of multivariate analysis with nine covariates-age, tumor location, tumor length, stage, pathological response, clinical response, MDM2 expression, p53 expression, and pRb expression, only pathological response and stage were significant factors for cause-specific survival. Conclusion: Expression of MDM2, p53, and pRb was not found to be clinically significant for predicting outcomes after CCRT in this study. Further studies with a larger patient population and longer follow-up periods are needed to re-evaluate the expression pattern and to identify new predictors for CCRT response.

Susceptibility of Two Potato Cultivars to Various Environmental Stresses (다양한 환경스트레스에 대한 감자 2품종의 감수성 분석)

  • Tang, Li;Kwon, Suk-Yoon;Sung, Chang-K;Kwak, Sang-Soo;Lee, Haeng-Soon
    • Journal of Plant Biotechnology
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    • v.30 no.4
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    • pp.405-410
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    • 2003
  • Environmental stress is the major limiting factor in plant productivity. In order to evaluate the stress tolerance of potato plants, leaf discs of two potato cultivars, Atlantic and Superior, were subjected to various stress conditions of high temperature, methyl viologen, H2O2, or $H_2O$$_2$. When potato leaf discs were exposed to high temperature at 37$^{\circ}C$ for 84 hr, Atlantic plants, a cultivar with high sensitivity to heat stress, showed about 20% higher membrane damage than Superior plants. When exposed to 2$\mu$M methyl violgen (MV), a superoxide generating non-selective herbicide, for 36 hr, Atlantic plants also showed about 38% higher membrane damage than Superior plants, and were more susceptible up to 10$\mu$M MV concentration tested. On treatment with 0.75M NaCl, Atlantic plants also had about 45% less chlorophyll contents in leaf discs than Superior plants. There was, however, no difference in chlorophyll content of two cultivars at higher NaCl concentrations. The effect of $H_2O$$_2$ on the two cultivars was mixed. At low $H_2O$$_2$ concentration (25 mM) , Superior plants were more susceptible to $H_2O$$_2$stress after 36 hr. However, at high $H_2O$$_2$ concentration (100 mM), Atlantic plants exhibited higher susceptibility after 36 hr. The results indicate that in vitro leaf discs reflecting the whole plants in this study will be useful for selection and characterization of elite transgenic potato plants with enhanced tolerance to environmental stress.