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Artificial Mutation for Silkworm Molecular Breeding Using Gene Scissors (유전자 가위의 이용과 누에 분자 육종을 위한 인위적 돌연변이 유발)

  • Hong, Jeong Won;Jeong, Chan Young;Yu, Jeong Hee;Kim, Su-Bae;Kang, Sang Kuk;Kim, Seong-Wan;Kim, Nam-Suk;Kim, Kee Young;Park, Jong Woo
    • Journal of Life Science
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    • v.30 no.8
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    • pp.701-707
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    • 2020
  • Gene editing technology using the clustered regularly interspaced short palindromic repeat (CRISPR) and the CRISPR associated protein (Cas)9 has been highly anticipated in developing breeding techniques. In this study, we discuss gene scissors as a tool for silkworm molecular breeding through analysis of Bombyx mori Kynurenine 3-Monooxygenase (BmKMO) gene editing using the CRISPR/Cas9 system and analysis of generational transmission through mutagenesis and selective crossing. The nucleotide sequence of the BmKMO gene was analyzed, and three guide RNAs (gRNAs) were prepared. Each synthesized gRNA was combined with Cas9 protein and then analyzed by T7 endonuclease I after introduction into the BM-N silkworm cell line. To edit the silkworm gene, K1P gRNA and Cas9 complexes were subsequently microinjected into the silkworm embryos; the hatching rate was 18% and the incidence of mutation was 60%. The gene mutation was verified in the heterozygous G0 generation, but no phenotypic change was observed. In homozygotes generated by self-crossing, a mutant phenotype was observed. These results suggest that silkworm molecular breeding using the CRISPR/Cas9 system is possible and could be an effective way of shortening the time required.

Creeping Bentgrass(Agrostis palustris Huds.) Control in Kentucky Bluegrass(Poa pratensis L.) Fairways (켄터키 블루그래스 페어웨이에서 문제가 되는 크리핑 벤트그래스 방제)

  • Tae Hyun-Sook
    • Asian Journal of Turfgrass Science
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    • v.19 no.2
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    • pp.65-72
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    • 2005
  • Creeping bentgrass (Agrostis palustris Huds.) had been the problematic weed for Kentucky bluegrass (Poa pratensis L.) fairway since it shows light green color all year. Experiment was carried out to determine the best herbicides combination to control creeping bentgrass in Kentucky bluegrass. fairway without injury. To investigate the efficacy of herbicides, five post-emergence herbicides of asulam WG ($87.6\%$), imazaquin SL ($20\%$), fenoxaprop-P-ethyl EC ($7\%$), mecoprop SL ($50\%$), triclopyr-TEA SL ($30\%$) and one pre-emergence herbicide pendimethalin EC ($31.7\%$) treated on 21 Sept. and 10 Nov. 2003. Kentucky bluegrass visual quality evaluated 30 and 50 days after application for phytotoxic effects of the herbicides. As a result, asulam WG (0.2g/$m^{2}$) and imazaquin SL (0.3ml/$m^{2}$) showed approximately $90\%$ of control in creeping bentgrass, but visual quality of Kentucky bluegrass significantly decreased from 20 to 50DAT (day after treatment). However, creeping bentgrass was acceptably controlled(over $80\%$) by fenoxaprop-P-ethyl EC (0.4ml/$m^{2}$)+triclopyr-TEA SL(0.3 ml/$m^{2}$) applied twice on 21 Sept. and 1 Oct. 2003 without serious injury on Kentucky bluegrass. Therefore, it is suggested that an application of fenoxaprop-P-ethyl EC (0.4ml/ $m^{2}$)+triclopyr-TEA SL (0.3 ml/$m^{2}$) may be more effective to control creeping bentgrass in Kentucky bluegrass with the least phytotoxicity by herbicides.

Optimal supervised LSA method using selective feature dimension reduction (선택적 자질 차원 축소를 이용한 최적의 지도적 LSA 방법)

  • Kim, Jung-Ho;Kim, Myung-Kyu;Cha, Myung-Hoon;In, Joo-Ho;Chae, Soo-Hoan
    • Science of Emotion and Sensibility
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    • v.13 no.1
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    • pp.47-60
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    • 2010
  • Most of the researches about classification usually have used kNN(k-Nearest Neighbor), SVM(Support Vector Machine), which are known as learn-based model, and Bayesian classifier, NNA(Neural Network Algorithm), which are known as statistics-based methods. However, there are some limitations of space and time when classifying so many web pages in recent internet. Moreover, most studies of classification are using uni-gram feature representation which is not good to represent real meaning of words. In case of Korean web page classification, there are some problems because of korean words property that the words have multiple meanings(polysemy). For these reasons, LSA(Latent Semantic Analysis) is proposed to classify well in these environment(large data set and words' polysemy). LSA uses SVD(Singular Value Decomposition) which decomposes the original term-document matrix to three different matrices and reduces their dimension. From this SVD's work, it is possible to create new low-level semantic space for representing vectors, which can make classification efficient and analyze latent meaning of words or document(or web pages). Although LSA is good at classification, it has some drawbacks in classification. As SVD reduces dimensions of matrix and creates new semantic space, it doesn't consider which dimensions discriminate vectors well but it does consider which dimensions represent vectors well. It is a reason why LSA doesn't improve performance of classification as expectation. In this paper, we propose new LSA which selects optimal dimensions to discriminate and represent vectors well as minimizing drawbacks and improving performance. This method that we propose shows better and more stable performance than other LSAs' in low-dimension space. In addition, we derive more improvement in classification as creating and selecting features by reducing stopwords and weighting specific values to them statistically.

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Effectiveness of Multimodal Pain Control in Early Phase After Arthroscopic Rotator Cuff Repair (관절경하 회전근 개 봉합술 후 다중 통증 조절법을 이용한 초기 통증 조절의 유용성)

  • Park, Chang-Min;Kim, Jong-Hae;Kim, Suk-Jun;Choi, Chang-Hyuk
    • Clinics in Shoulder and Elbow
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    • v.15 no.1
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    • pp.1-7
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    • 2012
  • Purpose: The purpose of this study was to identify the effectiveness of multimodal pain control method in an early phase after arthroscopic rotator cuff repair, under interscalene brachial plexus block, this study was performed. Materials and Methods: The study was progressed with the 80 cases of arthroscopic rotator cuff repair. Interscalene brachial plexus block was used to all of the 80 cases and patients were divided into 2 groups. Group A consisted of patients injected with bupivacaine, through subacromial space catheter after surgery, and group B consisted of patients with additional method of multimodal pain control using oral opioids, acetaminophen-tramadol complex and selective COX2 inhibitor. Subacromial cathter was removed after injection in both groups. The pain during the day time and night time was compared on the operation day, postoperative 1st, 2nd, 3rd day and 2nd weeks, and it was measured with VAS (visual analogue scale) score. Additionally, the number of ketolorac injection and side-effect related to analgesics was compared between the 2 groups. Results : The mean VAS score of night time on the operation day and day/night time pain of the 1st, 2nd, 3rd day and 2nd weeks was 7.4, 7.0/6.8, 4.5/5.2, 4.8/5.0, 2.2/2.7 on group A and 6.5, 4.3/5.4, 3.2/4.3, 3.0/4.1, 2.4/2.5 on group B, respectively. Significant difference was observed in the night pain on the operation day, 1st, 2nd, 3rd day time and 1st night time pain (p<.05). The average number of ketololac injection was 1.1 and 0.5 in each group, and there was no difference in the frequency of side effects. Conclusion: Multimodal pain control method, after arthroscopic rotator cuff repair, showed an effective early pain control and improved patients' satisfaction.

Biological Monitoring of Paint Handling Workers exposed to PAHs using Urinary 1-Hydroxypyrene (다핵방향족탄화수소류에 노출된 페인트 취급 근로자에서 요 중 1- Hydroxypyrene을 이용한 생물학적 모니터링)

  • Lee, Jong-Seong;Kim, Eun-A;Lee, Yong-Hag;Moon, Deog-Hwan;Kim, Kwang-Jong
    • Journal of Korean Society of Occupational and Environmental Hygiene
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    • v.15 no.2
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    • pp.124-134
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    • 2005
  • To investigate the exposure effect of polynuclear aromatic hydrocarbons (PAHs), we measured airborne total PAHs as an external dose, urinary 1-hydroxypyrene (1-OHP) as an internal dose of PAHs exposure, and analyzed the relationship between urinary 1-OHP concentration and PAHs exposure. The study population contained 44 workers in steel-pipe coating and paint manufacture industries. The airborne PAHs was obtained during survey day, and urine were sampled at the end of shift. Personal information on age, body weight, height, eniployment duration, smoking habit, and alcohol consumption was obtained by a structured questionnaire. Airborne PAHs were analyzed by the gas chromatograph with mass selective detector. Urinary 1-OHP levels were analyzed by the high performance liquid chromatograph with ultraviolet wavelength detector. For statistical estimation, t-test, ${\chi}^2$-test, analysis of variance, correlation analysis, arid regression analysis were executed by SPSS/PC (Windows version 10). The mean of environmental total PAHs was $87.8{\pm}7.81{\mu}g/m^3$. The mean concentration ($526.5{\pm}2.85{\mu}g/m^3$) of workers in steel-pipe coating industries using coal tar enamel was the higher than that ($17.5{\pm}3.36{\mu}g/m^3$) of workers in paint manufacture industries using coal tar paint. The mean of urinary 1-OHP concentration ($51.63{\pm}3.144{\mu}\;mol/mol$ creatinine) of workers in steel-pipe coating industries was the higher than that ($2.33{\pm}4.709{\mu}\;mol/mol$ creatinine) of workers in paint manufacture industries. The mean of urinary 1-OHP concentration of smokers was the higher than that of non-smokers. There was significant correlation between the urinary concentration of 1-OHP and the environmental concentration of PAHs (r=O.S48, p<0.001), pyrene(r=0.859, p<0.001), and urinary cotinine (r=0.324, p<0.05). The regression equation between the urinary concentration of 1-OHP in ${\mu}g/g$ creatinine($C_{1-OHP}$) and airborne concentration of PAHs (or pyrene) in ${\mu}g/m^3$ ($C_{PAHs}$ or Cpyrene) is: Log ($C_{1-OHP}$)=-0.650+0.889×Log($C_{PAHs}$), where $R^2=0.694$ and n=38 for p<0.001.Log ($C_{1-OHP}$)=1.087+0.707${\times}$Log(Cpyrene), where $R^2=0.713$ and n=38 for p<0.001. From the results of stepwise multiple regression analysis about 1-OHP, significant independents were total PAHs and urinary cotinine (adjusted $R^2=0.743$, p<0.001). In this study, there were significant correlation between the urinary concentration of 1-OHP and the airborne concentration of PAHs. The urinary 1-OHP was effective index as a biomarker of airborne PAHs in workplace. But it was influenced by non-occupational PAHs source, smoking.

Plant regeneration and transformation of grape (Vitis labrusca L.) via direct regeneration method (포도 (Vitis labrusca L.)의 직접 재분화 방법을 이용한 식물체 재분화와 형질전환)

  • Kim, Se Hee;Shin, Il Sheob;Cho, Kang Hee;Kim, Dae Hyun;Kim, Hyun Ran;Kim, Jeong Hee;Lim, Sun-Hyung;Kim, Ki Ok;Lee, Hyang Bun;Do, Kyung Ran;Hwang, Hae Seong
    • Journal of Plant Biotechnology
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    • v.40 no.4
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    • pp.210-216
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    • 2013
  • Efficient regeneration methods and transformation system are a priority for successful application of genetic engineering to vegetative propagated plants such as grape (Vitis labrusca L.). This research is to establish shoot regeneration system from plant explants for 'Campbell Early', 'Tamnara', 'Heukgoosul', 'Heukbosek' using two types of plant growth regulators supplemented to medium. The highest adventitious shoot regeneration rate of 5% was achieved on a medium containing of Murashige and Skoog (MS) inorganic salts and Linsmaier and Skoog (LS) vitamins, 2 mg/L of TDZ and 0.1 mg/L of IBA. Leaf tissue of 'Campbell Early', was co-cultivated with Agrobacterium strains, LBA4404 containing the vector pBI121 carrying with CaMV 35S promoter, gus gene as reporter gene and resistance to kanamycin as selective agent, the other Agrobacterium strains, GV3101 containing the vector pB7 WG2D carrying with mPAP1-D gene. mPAP1-D is a regulatory genes of the anthocyanin biosynthetic pathway. 'Campbell Early' harboring mPAP1-D gene was readily able to be selected by red color due to anthocyanin accumulation in the transformed shoot. These results might be helpful for further studies to enhance the transformation efficiency in grape.

Parental Survey for Children with Feeding Problems (섭취 문제가 있는 영유아 아동에 대한 부모설문조사)

  • Kim, Hye-Jin;Chung, Kyong-Mee;Park, Mi-Jung;Choe, Yon-Ho
    • Pediatric Gastroenterology, Hepatology & Nutrition
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    • v.11 no.2
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    • pp.179-186
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    • 2008
  • Purpose: Feeding problems are common for typically developing children and have negative effects on physical, behavioral, and cognitive development. The purpose of the current study was to examine factors related to childhood feeding problems for typically developing children via parental reports. Methods: The feeding questionnaire developed by the authors and the Childhood Eating Behavior Inventory (CEBI) were administered to 796 parents of children who recruited from 2 pediatric outpatient clinics (n=379) and community (n=417). Problem eaters (PE) were identified by parental report and frequency analysis was conducted for types of feeding problems and its severity, problem behaviors during mealtime, the feeding methods of the parents, and the need for treatment. Results: The CEBI scores were significantly different between the PE and non-problem eaters (NPE), which suggests that the parental reports were reliable. The younger children had more feeding problems than the older children. The most frequent and severe feeding problems were selective eating and longer mealtimes across all age groups. One-half of the children had more than one problem behaviors during mealtime. Most parents of PE used ineffective methods to deal with children's behaviors during meal time, such as cajoling, which was related to their increased level of stress. Many parents reported their children need professional help for the feeding problems. Conclusion: Feeding problems are prevalent among children, especially younger children. Children with feeding problems showed a number of problem behaviors during mealtime and parental coping methods appeared to be ineffective. The need for treatment was considerable, thus the characteristics of this population must be acknowledged for providing proper treatment and advice.

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Development of Practical Advanced Oxidation Treatment System for Decontamination of Soil and Groundwater Contaminated with Chlorinated Solvent (TCE, PCE) : Phase I (염소계 화합물(TCE, PCE)로 오염된 토양 및 지하수 처리를 위한 실용적 고도산화처리시스템 개발 (I))

  • Sohn, Seok-Gyu;Lee, Jong-Yeol;Jung, Jae-Sung;Lee, Hong-Kyun;Kong, Sung-Ho
    • Journal of Soil and Groundwater Environment
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    • v.12 no.5
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    • pp.105-114
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    • 2007
  • The most advanced oxidation processes (AOPs) are based on reactivity of strong and non-selective oxidants such as hydroxyl radical (${\cdot}OH$). Decomposition of typical DNAPL chlorinated compounds (TCE, PCE) using various advanced oxidation processes ($UV/Fe^{3+}$-chelating agent/$H_2O_2$ process, $UV/H_2O_2$ process) was approached to develop appropriate methods treating chlorinated compound (TCE, PCE) for further field application. $UV/H_2O_2$ oxidation system was most efficient for degrading TCE and PCE at neutral pH and the system could remove 99.92% of TCE after 150 min reaction time at pH 6($[H_2O_2]$ = 147 mM, UVdose = 17.4 kwh/L) and degrade 99.99% of PCE within 120 min ($[H_2O_2]$ = 29.4 mM, UVdose = 52.2 kwh/L). Whereas, $UV/Fe^{3+}$-chelating agent/$H_2O_2$ system removed TCE and PCE ca. > 90% (UVdose = 34.8 kwh/L, $[Fe^{3+}]$ = 0.1 mM, [Oxalate] = 0.6 mM, $[H_2O_2]$ = 147 mM) and 98% after 6hrs (UVdose = 17.4 kwh/L, $[Fe^{3+}]$ = 0.1 mM, [Oxalate] = 0.6 mM, $[H_2O_2]$ = 29.4 mM), respectively. We improved the reproduction system with addition of UV light to modified Fenton reaction by increasing reduction rate of $Fe^{3+}$ to $Fe^{2+}$. We expect that the system save the treatment time and improve the removal efficiencies. Moreover, we expect the activity of low molecular organic compounds such as acetate or oxalate be effective for maintaining pH condition as neutral. This oxidation system could be an economical, environmental friendly, and practical treatment process since the organic compounds and iron minerals exist in nature soil conditions.

The Validity of a Combined High Saphenous Division and Sclerotherapy for Varicose Vein (하지정맥류의 치료에 있어서 복재정맥 분리 결찰 및 혈관 경화요법 병용의 유용성)

  • Choi Se-Yong;Yang In-Suk;Won Tae-Hee
    • Journal of Chest Surgery
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    • v.39 no.7 s.264
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    • pp.544-548
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    • 2006
  • Background: The purpose of this study is to evaluate the validity of a combined high saphenous vein division and sclerotherapy without sacrificing saphenous vein for varicose vein. Material and Method: Between August 2004 and October 2005, 70 limbs in 50 patients were treated by a combined high saphenous vein division and sclero-therapy. The operative indication is valvular incompetence of femoral-saphenous or popliteal-saphenous junction. Patients received local anesthesia and were treated by a combined high saphenous vein division, ligation of incompetent perforating vein and $1{\sim}3%$ sclerosant. Patients received 1 day hospitalization and applied in com-pression stocking for 6 weeks. Patients followed after 1 week, 1 and 3 months. Result: Mean age of patients was $50{\pm}11$. The female was more common. 8 patients was no symptom, another 42 patients complained of pain, heaviness and fatigue of limbs. The symptoms of varicose vein disappeared 1 month after the procedure in all symptomatic patients. 8 patients needed a adjuvant sclerotherapy for residual varicose vein on 1 week after the procedure. There were only minor complications such as hematoma (1), wound infection (1), thrombophlebitis (20), skin blister (10), hyper-pigmentation (1), and skin ulcer (1). Conclusion: We concluded that a combined high saphenous vein division and sclerotherapy without sacrificing saphenous vein for varicose vein is simple, less invasive, economical, and effective treatment for primary varicose vein, and it has a special advantage that saphenous vein can be used as a bypass conduit later. The method was selective in old aged patients.

20-Year Experience of Surgical Treatment for Postpneumonectomy Empyema (전폐절제술 후 사강에 발생한 농흉의 치료)

  • 김형렬;김영태;성숙환;김주현
    • Journal of Chest Surgery
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    • v.35 no.7
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    • pp.542-547
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    • 2002
  • Background: Postpneumonectomy empyema(PPE) is an infrequent but potentially life-threatening complication. To date, various surgical efforts have been made to manage this complication. We reviewed our 20-year surgical experience of PPE and long-term follow-up data. Material and Method: Total of 37 patients who were treated for PPE between fan, 1980 and Jun, 2000 were included. Various clinical factors such as micro-organism, operative method and timing, presence of bronchopleural fistula(BPF), underlying disease and fate of empyema cavity were retrospectively reviewed and analyzed. Result: Majority of patients(34) underwent Eloesser operation for effective drainage. There was only one operative mortality. The causative organisms were Staphylococcus species and Pseudomonas species in 46% BPF was found in 20 cases, among which spontaneous closures took place in 4 cases. The chest wall was closed in 40%(8/20) of patients with BPF, compared to 59%(10/17) without BPF. The closure rate was statistically better in patients without BPF(p=0.006). Even though the patients with benign disease showed higher closure rate(50%) than those with lung cancer (31%), the difference was not significant(p=0.25). Conclusion: Eloesser procedure was an effective method for initial drainage of PPE cavity with low operative mortality. Given the findings of low spontaneous closure rate of BPF, aggressive approach to close the BPF is mandatory to achieve the final goal of chest wall closure. It was found that majority of patients still left their chest cavity opened, even after controlling the active inflammation of the empyema cavity. More aggnessive approach for chest wall closure is recommended in all patents with benign disease and in selective patients with lung cancer if there is no evidence of recurrence at several years after the initial operation.