• Title/Summary/Keyword: point quadruple

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Cloning and Site-Directed Mutagenesis of Musca domestica Acetylcholinesterase for Enhancing Sensitivity to Organophosphorus and Carbamate Insecticides

  • Kim, Chung-Sei;Kim, Su-Il
    • Journal of Microbiology and Biotechnology
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    • v.16 no.11
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    • pp.1760-1772
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    • 2006
  • Mature acetylcholinesterase (AChE) gene (gm, 1,836 bp) was cloned from the housefly and successfully expressed in the E. coli CodonPlus (DE3) RIL system (GM-E, 72 kDa) with a yield of 1,630 mU/g fresh cells. Using the gm, 10 kinds of mutants were constructed and expressed for enhancing sensitivity to insecticides. The sensitivity of these mutants to five kinds of organophosphate (OP) and three carbamate insecticides was investigated by measuring the apparent bimolecular inhibition constant ($k_i=k_2/K_d$). Surprisingly, the sensitivity of quadruple mutant IGFT was enhanced as much as 7-fold for acephate, 164-fold for demeton-S-methyl, 484-fold for dichlorvos, 523-fold for edifenphos, 30-fold for ethoprophos, 30-fold for benfuracarb, 404-fold for carbaryl, and 107-fold for furathiocarb, compared with that of GM-E, although the sensitivity of each single point mutant was slightly increased. These mutational studies indicated that (i) contradictory to Walsh et al. [39], the residue 327 is the important key residue for enhancing sensitivity as much as the residue 262, (ii) the residue 82 and additional residues of 234, 236, and 585 are also important, and (iii) sensitivity was cooperatively accelerated as the number of strategic mutations increased.

Comparison of Scapular Position Between Operation and Non-operation Side to the Rotator Cuff Surgery (회전근개 수술 환자의 수술측과 비수술측 간에 견갑골 자세 비교)

  • Jeong, Eui-young;Kim, Suhn-yeop
    • The Journal of Korean Academy of Orthopedic Manual Physical Therapy
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    • v.22 no.2
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    • pp.15-20
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    • 2016
  • Background: The purpose of this study was to compared of scapular position between operation side and non-operation side to the rotator cuff surgery. Methods: This study was carried out with a total 34 patients: male (n=14), female (n=20). Shoulder range of motion (ROM), the quadruple visual analogue scale (QVAS), the shoulder pain and disability index (SPADI), and the scapular index (SI) were used to assess shoulder posture and function. SI was the resting position of the scapular was determined by measuring the distance from the mid-point of the sternal notch (SN) to the medial aspect of the coracoid process (CP) and the horizontal distance from the posterolateral angle of the acromion (PLA) to the thoracic spine (TS) with a soft tape measure. The SI was calculated using the equation: [(SN to CP/PLA to TS) ${\times}$ 100]. Results: There were no significant difference in ROM, QVAS to rotator repair patients according to SI (p>.05). There were significant differences in SI between the operation side and the non-operation side (p<.01). Conclusions: Scapular position was operation side more internal rotation, protraction, abduction than non-operation side. Therefore, health professionals managing for rotator cuff tear repair patients should consider scapular position.

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The Study on the Contents and Satisfaction of Oriental Medicine Examination Program (한방건강검진 프로그램의 내용과 참가자들의 만족도에 관한 연구)

  • Lee Eun-Kyoung;Jahng Doo-Sub;Song Yung-Sun;Lee Ki-Nam
    • Journal of Society of Preventive Korean Medicine
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    • v.6 no.1
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    • pp.51-95
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    • 2002
  • 1. As a point at issue of occidental medicine examination, followings have been indicated, 1) Occidental medical examination is performed by mainly test and instrument and therefore, role of doctors could be excluded and it could be diagnosed only for target disease and 2) in Korea, it could not be conducted in public medical system and therefore improvement effect of public health promotion could not be made due to increase of total medical expenses and infirmity of post management. These points are substantial limit of paradigm resided in occidental medicine and also problem caused by unique characteristics of medical system of Korea. In Korea, result of occidental medical examination show increased health risk from aged thirties and major diagnosed diseases are circulatory disease or chronic disease such as hepatic, hyperlipemic, hypertensional disease and diabetes, etc. Accounting that those chronic disease make a role as major public health risk, it is difficult that medical examination by only occidental medicine make effect on public health promotion. 2. Characteristics of oriental medicine examination could be summarized as diagnosis (information acquisition based on the facts) and demonstration (speculation based on acquired information) and in addition, quadruple diagnosis, as a medical examination method, include test as well as basic examination. Accounting on oriental medical examination, principally it is performed by independent herbalist and therefore, herbalist could acquire systemic result during first medical examination. Based on the theory of inner-outer examination, oriental medical examination has a principle of universe theory, systemic analysis of quadruple diagnosis, demonstration & reasoning. In addition, root of oriental medical examination could be found in pre-disease theory, a principal theory of oriental medicine. Pre-disease service could prove the advantage of oriental medicine in medical examination activities and therefore, it is needed that content of oriental medical examination should be actualized in current medical system. 3. In this study, oriental medical examination program, comprised of pulse-diagnosis, contrast muscular taking and medical consultation of herbalist is progressed communally with occidental medical examination. As pre-examination, questionary was given of general characteristics, health promoting life style, physical constitution and subjective symptom of musculoskeletal system. In addition, post-examination notification was given to subjects about health promoting control, physical constitution regimen and management of musculoskeletal system. During this study program, verification was conducted for input of acquired information and difference of each information after analysis and in addition, performed was analysis of factor influencing health promoting life style and musculoskeletal subjective symptom and evaluation of relationship of physical constitution and health promoting life study. In addition, it was verified of difference between musculoskeletal subjective symptom and result of muscle contrast picture evaluation. 4. Evaluation of oriental medical examination model is divided into 2 categories of oriental medical examination-consultation and result evaluation -post management. Oriental medical examination-consultation demands establishment of examination system, standardization of examination and establishment of examination form and in addition, it should be admitted as enlarged examination assists systemic quadruple diagnosis of herbalist not a key of oriental medical examination. In addition, information acquisition for research purpose should be performed according to the systemic research plan based on the separation of questionary for examination purpose and research purpose. For evaluation of the result, it was concluded that needed are result evaluation meets oriental medical system and post-notification system, informing health management information, based on examination result. 5. Accounting on satisfaction for oriental medical examination model, affirmative reply was much higher (66%) than negative (8.64%). Satisfaction of each area was in order of consultation of herbalist, systemic muscle contrast taking, pulse examination, post-notification and questionary fill-up and dissatisfaction was in order of post-notification, questionary fill-up, consultation of herbalist, systemic muscle contrast taking and pulse measurement. Satisfaction for collaborative examination of occidental and oriental medicine was over 60% and over 75% hope oriental examination would be included in later medical examination program. Based on this result, collaborative examination including occidental and oriental medicine could increase satisfaction of subjects for medical examination program.

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Effects of Temperature and Light Intensity on the Growth of Red Pepper(Capsicum annuum L.) in Plastic House During Winter. I. Fluctuations of Temperature and Light Environment in the Multilayered Plastic House Grown Red Pepper (동계 Plastic house내 고추(Capsicum annuum L.) 육묘시 온도와 광도가 생장에 미치는 영향 I. 다중피복 고추육묘 시설내의 온도 및 광환경 영향)

  • 정순주;이범선;권용웅
    • Journal of Bio-Environment Control
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    • v.3 no.2
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    • pp.106-118
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    • 1994
  • This study was conducted to analyze the effects of fluctuations in temperature, light intensity and soil temperature on the growth of red pepper seedlings in the nonheated plastic houses with various number of layers and in the open field. Relationship between the optimal environment and the growth of seedlings was discussed, and the maximum and minimum outdoor temperatures in Kwangju area from 1941 to 1985 were analyzed. The results obtained were as follows; 1. The minimum temperature in tunnel with quadruple coverings of P. E. film from December 20 to February 25 was decreased to 5$^{\circ}C$ mostly, where the exposure to chilling temperature could not be avoided during this period. The maximum temperature was increased to 33$^{\circ}C$ mostly and 42$^{\circ}C$ in peak, where some ventilation was needed. 2. The diurnal differences of inside temperature, increasing with number of layers, were 16 to 38$^{\circ}C$, while those of outside temperature were 5 to 1$0^{\circ}C$. 3. The cold injury in the quadruple coverings during winter occurred all the times below 12$^{\circ}C$ and as many as 200 times over 3$0^{\circ}C$, while effectiveness of thermal insulation in the multilayered nonheating plastic houses were clearly proved. 4. The inside light intensity was markedly reduced with the increment of layers and the minimum light intensity fallen down below the light compensation point for the growth of red pepper plants regardless of the number of layers. 5. Until 10 a. m., the temperature in the daytime during December 20 to mid - February showed below 10 to 12$^{\circ}C$ which was the limiting temperature for the growth of red pepper seedlings. After 4 p. m., the light intensity was sharply reduced despite of the air temperature kept over 12$^{\circ}C$. Therefore, limiting factors for the growth of red pepper seedlings were the temperature before 10 a. m. and the light intensity after 4 p. m. 6. The minimum soil temperature in quadruple coverings showed around 1$0^{\circ}C$ where the physiological damage for red pepper seedlings might be occurred. 7. The minimum outdoor temperatures from 1941 to 1985 was -19.4$^{\circ}C$, observed in the 5th January.

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Single-Base Genome Editing in Corynebacterium glutamicum with the Help of Negative Selection by Target-Mismatched CRISPR/Cpf1

  • Kim, Hyun Ju;Oh, Se Young;Lee, Sang Jun
    • Journal of Microbiology and Biotechnology
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    • v.30 no.10
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    • pp.1583-1591
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    • 2020
  • CRISPR/Cpf1 has emerged as a new CRISPR-based genome editing tool because, in comparison with CRIPSR/Cas9, it has a different T-rich PAM sequence to expand the target DNA sequence. Single-base editing in the microbial genome can be facilitated by oligonucleotide-directed mutagenesis (ODM) followed by negative selection with the CRISPR/Cpf1 system. However, single point mutations aided by Cpf1 negative selection have been rarely reported in Corynebacterium glutamicum. This study aimed to introduce an amber stop codon in crtEb encoding lycopene hydratase, through ODM and Cpf1-mediated negative selection; deficiency of this enzyme causes pink coloration due to lycopene accumulation in C. glutamicum. Consequently, on using double-, triple-, and quadruple-base-mutagenic oligonucleotides, 91.5-95.3% pink cells were obtained among the total live C. glutamicum cells. However, among the negatively selected live cells, 0.6% pink cells were obtained using single-base-mutagenic oligonucleotides, indicating that very few single-base mutations were introduced, possibly owing to mismatch tolerance. This led to the consideration of various target-mismatched crRNAs to prevent the death of single-base-edited cells. Consequently, we obtained 99.7% pink colonies after CRISPR/Cpf1-mediated negative selection using an appropriate single-mismatched crRNA. Furthermore, Sanger sequencing revealed that single-base mutations were successfully edited in the 99.7% of pink cells, while only two of nine among 0.6% of pink cells were correctly edited. The results indicate that the target-mismatched Cpf1 negative selection can assist in efficient and accurate single-base genome editing methods in C. glutamicum.