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Molecular Characterization of Cinnamate 4-Hydroxylase gene in Red Hot Pepper (Capsicum annuum L.) (고추에서 분리한 Cinnamate 4-Hydroxylase 유전자의 분자생물학적 특성)

  • Kim Kye-Won;Ha Sun-Hwa;Cho Kang-Jin;Kim Eun-Ju;Lee Min-Kyung;Yu Jae-Ju;Kim Jong-Guk;Lee Shin-Woo
    • Journal of Plant Biotechnology
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    • v.32 no.3
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    • pp.167-173
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    • 2005
  • Three different cDNAS for cinnamate 4-hydroxylase (C4H) which are involved in the second step of the general phenylpropanoid pathway were isolated and designated as pc4h1 (1,755 bp), pc4h2 (1,655 bp), and pc4h3 (1,316 bp), respectively. The nucleotide sequence analysis revealed that both pc4h1 and pc4h2 clones encode polypeptides of 505 amino acids frame but pc4h3 clone was truncated at the 5'-end of coding region. The alignment of the deduced amino acid sequences showed that PC4H1 and PC4H2 are highly homologous (95.8% identical) with each other and contain three conserved domains which are typical in cytochrome P450 monooxygenase: proline-rich region, threonine-containing binding pocket for the oxygen molecule, and heme binding region. In addition, result of the phylogenic tree analysis revealed that both pepper C4Hs belong to Class 1. pc4h2 transcription was strongly induced in wounded fruit (400%) and root (200%) relative to its very low basal level but not in leaf or stem tissue. In case of pc4h1, the basal level of transcription was higher than pc4h2 but induction by wounding was lower in fruit and root while leaf and stem tissues did not respond to wounding. The basal level of pc4h3 transcripts was not, if any, detectable and response to wounding was not observed.

Multiplex PCR method for environmental monitoring of approved LM cotton events in Korea (국내 승인 LM면화의 자연환경 모니터링을 위한 multiplex PCR 개발)

  • Jo, Beom-Ho;Seol, Min-A;Shin, Su Young;Kim, Il Ryong;Choi, Wonkyun;Eum, Soon-Jae;Song, Hae-Ryong;Lee, Jung Ro
    • Journal of Plant Biotechnology
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    • v.43 no.1
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    • pp.91-98
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    • 2016
  • The growth area of living modified (LM) cotton has steadily increased every year, since its first commercialization in 1996. Development of environmental risk assessment tools and techniques for LM cotton is required for ecosystem safety. We therefore developed multiplex PCR assays for simultaneous detection of two (MON15985, MON531) and four (GHB614, LLCOTTON25, MON88913 and MON1445) LM cotton events approved in Korea, with event specific primer pairs. The PCR reactions were optimized by using event specific primers of six LM cottons at various concentrations. The reactions allows amplification of estimated amplicons of MON15985 (214 bp), MON531 (270 bp), GHB614 (119 bp), LLCOTTON25 (164 bp), MON88913 (276 bp), and MON1445 (389 bp) from multiplex PCR reactions. The multiplex PCR assay developed allowed that two annealing steps (15 cycles at $55^{\circ}C$ and 25 cycles at $60^{\circ}C$) were performed for amplification of distinguished two LM cottons, and only one annealing step (50 cycles at $60^{\circ}C$) was necessary for tetraplex PCR. Primer extension step of all PCR reactions was skipped for time-effective amplification. Our methods suggest that two multiplex PCR assays can be cost-effective and a rapid diagnostic tool for environmental LMO monitoring of six LM cottons.

Establishment of Featal Heart Surgery with an Improvement of the Placental Blood Flow in Cardiopulmonary Bypass Using Fetal Lamb Model (양태아를 이용한 심폐우회술에서의 태반혈류개선을 통한 태아심장수술의 기반기술 확립)

  • 이정렬;박천수;임홍국;배은정;안규리
    • Journal of Chest Surgery
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    • v.37 no.1
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    • pp.11-18
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    • 2004
  • Background: We tested the effect of indomethacine and total spinal anesthesia on the improvement of placental flow during cardiopulmonary bypass on fetal lamb. Material and Method: Twenty fetuses at 120 to 150 days of gestation were subjected to bypass via trans-sternal approach with a 12 G pulmonary arterial cannula and 14 to 18 F venous cannula for 30 minutes. All ewes received general anesthesia with ketamine. In all the fetuses, no anesthetic agents were used except muscle relaxant. Ten served as a control group in which placenta was worked as an oxygenator during bypass (Control group). The remainder worked as an experimental group in which pretreatment with indomethacine and total spinal anesthesia was performed before bypass with the same extracorporeal circulation technique as control group (Experimental group). Observations were made every 10 minutes during a 30-minute bypass and 30-minute post bypass period. Result: Weights of the fetuses ranged from 2.2 to 5.2 kg. In Control group, means of arterial pressure decreased from 44.7 to 14.4 mmHg and means of Pa$CO_2$ increased from 61.9 to 129.6 mmHg at each time points during bypass. Flow rate was suboptimal (74.3 to 97.0 $m\ell$/kg/min) during bypass. All hearts fibrillated immediately after the discontinuation of bypass. On the contrary, in Experimental group, means of arterial pressure reamined higher (45.8 to 30 mmHg) during bypass (p<0.05). Means of Pa$CO_2$ were less ranging from 59.8 to 79.4 mmHg during bypass (P<0.05). Flow rates were higher (78.8 to 120.2 $m\ell$/kg/min) during bypass (p<0.05). There were slower deterioration of cardiac function after cessation of bypass. Conclusion: In this study, we demonstrated that the placental flow was increased during fetal cardiopulmonary bypass in the group pretreated with indomethacine and total spinal anesthesia. However, further studies with modifications of the bypass including a creation of more concise bypass circuit, and a use of axial pump are mandatory for the clinical application.

Significance of p53 as a Prognostic Factor in Non-Small Cell Lung Carcinoma (비소세포 폐암종에 있어서 p53의 예후 인자로서의 의의)

  • 이상호;한정호;김관민;김진국;심영목;장인석
    • Journal of Chest Surgery
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    • v.37 no.8
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    • pp.672-683
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    • 2004
  • Background: The treatment results of the advanced lung carcinoma is not satisfactory with the present therapeutic modalities: surgical resection, anti-cancer chemotherapy, and radiotherapy or combination therapy. To predict the prognosis of the non-small-cell lung carcinoma, TNM classification has been was as the basic categorization; however, it has been not satisfactory. It is necessary to consider the causes and the prognosis of the lung carcinoma from another points of view rather the conventional methods. We intended to find out the relationship between the major apoptotic factor, p53 gene and the prognosis of the patient with lung carcinoma. Material and Method: Three hundreds and fifty-nine patients with lung carcinoma who underwent surgery were analysed. We observed p53 protein accumulated in the cellular nuclei. The p53 protein was detected by immuno-histo-chemical method. We collected information of the patient retrospectively. Result: p53 protein densities were observed in 40% in average as a whole. The protein density was 44 percent in man, 25 percent in woman, 49 percent in the squamous cell carcinoma, and 38 percent in the adenocarcinoma. There were significant correlations between the p53 protein density and the mortality in the squamous cell carcinoma (p=0.025), follow-up duration in TNM stage I group (p=0.010), and follow-up duration in the lobectomy patient group (p=0.043), and tumor cell differentiation (p=0.009). p53 protein densities were significantly different between the lobectomy and the pneumonectomy group (p=0.044). Conclusion: The authors found that p53 protein had some correlations with the prognosis of the lung cancer partially in some factors. We suggest the p53 protein density could be used as a marker of prognosis in the non-small-cell lung carcinoma.

The Evaluation of Usefulness of Pixelated Breast-Specific Gamma Imaging in Thyroid scan (Pixelated Breast-Specific Gamma Imaging(BSGI) 감마 카메라를 이용한 갑상선 검사의 유용성 평가)

  • Jung, Eun-Mi;Seong, Ji-Hye;Yoo, Hee-Jae
    • The Korean Journal of Nuclear Medicine Technology
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    • v.15 no.1
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    • pp.90-93
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    • 2011
  • Purpose: A Pixelated BSGI gamma camera has features to enhance resolution and sensitivity and minimize the distance between detector and organs by narrow FOV. Therefore, it is known as useful device to examine small organs such as thyroid, parathyroid and gall bladder. In general, when we would like to enlarge the size of images and obtain high resolution images by gamma camera in nuclear medicine study, we use pinhole collimator. The purpose of this study is to evaluate the usefulness of Pixelated BSGI gamma camera and to compare to it using pinhole collimator in thyroid scan which is a study of typical small organs. Materials and methods: (1) The evaluation of sensitivity and spatial resolution: We measured sensitivity and spatial resolution of Pixelated BSGI with LEHR collimator and Infinia gamma camera with pinhole collimator. The sensitivity was measured by point source sensitivity test recommended by IAEA. We acquired images considering dead time in BSGI gamma camera for 100 seconds and used $^{99m}TcO4-\;400{\mu}Ci$ line source. (2) The evaluation of thyroid phantom: The thyroid phantom was filled with $^{99m}TcO4-$. After set 300 sec or 100 kcts stop conditions, we acquired images from both pixelated BSGI gamma camera and Infinia gamma camera with LEHR collimator. And we performed all thyroid studies in the same way as current AMC's procedure. Results: (1) the result of sensitivity: As a result, the sensitivity and spatial resolution of pixelated BSGI gamma camera were better than Infinia's. The sensitivities of pixelated BSGI and Infinia gamma camera were $290cps/{\mu}Ci$ and $350cps/{\mu}Ci$ respectively. So, the sensitivity of pixelated BSGI was 1.2 times higher than Infinia's (2) the result of thyroid phantom: Consequently, we confirmed that images of Pixelated BSGI gamma camera were more distinguishable between hot and cold spot compared with Infinia gamma camera. Conclusion: A pixelated BSGI gamma camera is able to shorten the acquisition time. Furthermore, the patients are exposed to radiation less than before by reducing amount of radiopharmaceutical doses. Shortening scan time makes images better by minimizing patient's breath and motion. And also, the distance between organ and detector is minimized because detector of pixelated BSGI gamma camera is small and possible to rotate. When patient cannot move at all, it is useful since device is feasible to move itself. However, although a pixelated BSGI gamma camera has these advantages, the effect of dead time occurs over 2000 cts/s since it was produced only for breast scan. So, there were low concentrations in organ. Therefore, we should consider that it needs to take tests to adjust acquisition time and amount of radiopharmaceutical doses in thyroid scan case with a pixelated BSGI gamma camera.

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Seasonal Variations of Environmental Factors and Distribution of Anabaena cylindrica Growth-Inhibiting Bacteria in the Lower Daechung Reservoir (대청호 하류에서 환경요인과 Anabaena cylindrica 생장억제세균의 계절별 분포 변화)

  • Lee, Jung-Ho;Kim, Chul-Ho
    • Korean Journal of Ecology and Environment
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    • v.33 no.2 s.90
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    • pp.128-135
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    • 2000
  • The authors surveyed the seasonal variations of environmental factors, the distributions of heterotrophic bacteria and Anabaena cylindrica growth-inhibiting bacteria at each water layer in Daechung Reservoir to verify the role of bacteria during the extinction of bloom. Average water depth at site 1, 2, and 3 were 25.5 m, 15.0 m and 12.3 m, respectively. Water temperature showed a typical pattern seasonally. The variation of DO was relatively inverse proportional to that of water temperature, although it was irregular during summer time. DO decreased gradually to early May, fluctuated sharply after then, and followed by gradual increasement after middle of September. This variation pattern was notable at surface layer. There was remarkable difference in DO concentraion between surface layer and the other water layers during the period in which DO irregulary varied. The variation range of chlorophyll-a concentraion at surface layer in summer time was broad, and it was relatively high when DO was high. The population size of heterotrophic bacteria was high from Spring to Autumn, an declined after September when the water temperature droped rapidly. Especially this variation pattern was prominent at the surface layer. Bacteria that inhibit the growth of A. cylindrica was almost not detected by June, and its distribution increased in July. Afterward, it showed different variation pattern between each site. The distribution of A. cylindrica growth-inhibiting bacteria was higher at the middle and bottom layer than the surface layer in July and October, when it was larger at all sites for the study period. This result suggests that the antagonistic bacteria exhibit higher activity when host activity drops. These results also suggest that natural water bacteria control the distirbution of cyanobacteria, especially its activity as controller is remarkable when cyanobacterial growth declines.

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A review of factors that regulate extracellular enzyme activity in wetland soils (습지 토양 내 체외효소 활성도를 조절하는 인자에 대한 고찰)

  • Kim, Haryun
    • Korean Journal of Microbiology
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    • v.51 no.2
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    • pp.97-107
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    • 2015
  • Wetlands constitute a transitional zone between terrestrial and aquatic ecosystems and have unique characteristics such as frequent inundation, inflow of nutrients from terrestrial ecosystems, presence of plants adapted to grow in water, and soil that is occasionally oxygen deficient due to saturation. These characteristics and the presence of vegetation determine physical and chemical properties that affect decomposition rates of organic matter (OM). Decomposition of OM is associated with activities of various extracellular enzymes (EE) produced by bacteria and fungi. Extracellular enzymes convert macromolecules to simple compounds such as labile organic carbon (C), nitrogen (N), phosphorus (P), and sulfur (S) that can be easily taken up by microbes and plants. Therefore, the enzymatic approach is helpful to understand the decomposition rates of OM and nutrient cycling in wetland soils. This paper reviews the physical and biogeochemical factors that regulate extracellular enzyme activities (EEa) in wetland soils, including those of ${\beta}$-glucosidase, ${\beta}$-N-acetylglucosaminidase, phosphatase, arylsulfatase, and phenol oxidase that decompose organic matter and release C, N, P, and S nutrients for microbial and plant growths. Effects of pH, water table, and particle size of OM on EEa were not significantly different among sites, whereas the influence of temperature on EEa varied depending on microbial acclimation to extreme temperatures. Addition of C, N, or P affected EEa differently depending on the nutrient state, C:N ratio, limiting factors, and types of enzymes of wetland soils. Substrate quality influenced EEa more significantly than did other factors. Also, drainage of wetland and increased temperature due to global climate change can stimulate phenol oxidase activity, and anthropogenic N deposition can enhance the hydrolytic EEa; these effects increase OM decomposition rates and emissions of $CO_2$ and $CH_4$ from wetland systems. The researches on the relationship between microbial structures and EE functions, and environmental factors controlling EEa can be helpful to manipulate wetland ecosystems for treating pollutants and to monitor wetland ecosystem services.

Simulation of Natural Air Layer Drying of Rough Rice (시뮬레이숀에 의한 벼의 상온통풍층건조방법(常温通風層乾燥方法)에 관(關)한 연구(硏究))

  • Park, Jae Bok;Koh, Hak Kyun;Chung, Chang Joo
    • Journal of Biosystems Engineering
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    • v.8 no.1
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    • pp.47-60
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    • 1983
  • 상온통풍(常溫通風)을 이용(利用)한 In-bin drying에 대(對)한 많은 실험결과(實驗結果)에 의(依)하면 우리나라 10월(月)의 기상조건(氣象條件)은 저온건조(低溫乾燥) system에 적합(適合)한 건조능력(乾燥能力)을 가지고 있는 것으로 나타났다. 최근(最近) Computer를 이용(利用)한 Simulation model이 개발(開發)되어 건조현상(乾操現象)에 관(關)한 경제적(經濟的)이고 효율적(效率的)인 분석(分析)이 가능(可能)하게 되었다. 이러한 분석결과(分析結果)에 의(依)하면 초기함수율(初期含水率)이 높은 벼를 Full-bin을 이용(利用)한 상온통풍건조(常溫通風乾操)를 할 경우(境遇) 건조기간(乾燥期間)이 길어지며 bin내(內)의 상층부(上層部) 곡물(穀物)이 변질(變質)되는 문제점(問題點)이 발생(發生)하였다. 또한 벼의 수확작업체계(收穫作業體系)가 관행(慣行) 및 Binder작업체계(作業體系)에서 점차(漸次) Combine작업체계(作業體系)로 전환(轉換)되어감에 따라 포장(圃場)에서의 건조(乾燥)가 어려우며 예취(刈取), 탈곡작업과정(脫穀作業過程)에서의 기계적(機械的)인 곡물(穀物) 손실(損失)을 줄이기 위(爲)하여 함수율(含水率)이 비교적(比較的) 높은 벼를 수확(收穫)하여야 한다. 본(本) 연구(硏究)의 목적(目的)은 상온통풍건조(常溫通風乾燥)에 있어서 건조능력(乾燥能力)을 증가(增加)시키기 위(爲)하여 곡물(穀物)을 일정기간(一定期間) 나누어서 bin에 넣고 건조(乾燥)를 하는 Layer drying의 Simulation model을 개발(開發)하고 이 Model에 수원지방(水原地方)의 7년간(年間) 평균(平均) 기상자료(氣象資料)를 입력(入力)시켜 곡물(穀物)의 초기함수율(初期含水率), 투입량(投入量), 투입기간(投入期間)의 변화(變化)에 따른 Layer drying현상(現象)을 규명(糾明)하는데 있다. Simulation에 사용(使用)된 bin의 크기는 직경(直徑) 2m, 깊이 1.5m이며 송풍(送風)팬의 용량(容量)은 0.5HP이었다. Simulation분석(分析) 결과(結果)를 요약(要約)하면 다음과 같다. (1) Layer drying의 Simulation model을 개발(開發)하였으며 이 Model은 벼의 상온통풍건조(常溫通風乾燥) 실험(實驗)에서 함수량(含水量) 변화(變化)의 이론치(理論値)와 실제실험치(實際實驗値)가 잘 일치(一致)하였다. (2) 곡물투입기간(穀物投入期間) 1일(日)을 Full-bin drying으로 간주(看做)할 때 Layer drying의 건조성능(乾燥性能)은 Full-bin보다 높은 것으로 나타났다. (3) 연속송풍(連續送風)(24시간(時間))을 할 경우(境遇) 곡물투입기간(穀物投入期間)이 증가(增加)함에 따라 건조기간(乾燥期間)의 감소경향(減少傾向)은 단속송풍(斷續送風)인 경우(境遇)보다 적었지만 건조기간(乾燥期間)은 단축(短縮)되었다. 그러나 건조(乾燥)에너지의 소모(消耗)는 단속송풍(斷續送風)일 때보다 크게 나타났다. (4) 단속송풍(斷續送風)(9 : 00AM~6 : 00PM)일 경우(境遇) 곡물투입기간(穀物投入期間)을 증가(增加)시키면 건조기간(乾燥期間)이 크게 줄어 들었다. (5) 곡물(穀物)의 초기함수율(初期含水率)이 21% 이하(以下)일 경우(境遇) 연속(連續) 및 단속송풍(斷續送風)에서 건조기간(乾燥期間)의 차이(差異)가 별로 없었다. (6) 곡물(穀物)의 초기함수율(初期含水率)이 높으면 Full-bin drying에서는 상부층(上部層)에 곡물(穀物)이 변질(變質)될 우려(憂慮)가 있으나 Layer drying에서는 곡물투입량(穀物投入量)을 조절(調節)하면 이것을 방지(防止)할 수 있었다. (7) 건조(乾燥)가 완료(完了)된 후(後) 층별(層別) 최종곡물(最終穀物) 함수율(含水率)은 모든 건조조건(乾燥條件)에서 동일(同一)하였으나 bin의 하부층(下部層)은 과건조(過乾燥) 물상(物象)을 일으켰다.

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Treatment Level of a Pond System for Ecological Treatment and Recycling of Animal Excreta (생태적 축산폐수 처리 및 재활용 연못시스템의 폐수처리수준)

  • Yang, Hong-Mo;Rhee, Chong-Ouk
    • Korean Journal of Environmental Agriculture
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    • v.17 no.1
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    • pp.70-75
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    • 1998
  • A model of pond system is developed for treatment and recycling of excreta from twenty-five adult dairy cattle. It is composed of wastewater treatment ponds and small fish ponds. Those are three facultative ponds in series; primary-secondary-tertiary pond and these are designed to rear carps without feeding. A pit is constructed at the bottom of primary pond for efficient sludge sedimentation and effective methane fermentation. It is contrived to block into it the penetration of oxygen dissolved in the upper layer of pond water. The excreta from the cattle housed in stalls are diluted by water used for clearing them. The washed excreta flow into the pit. The average yearly $BOD_5$ concentration of influent is 398.7mg/l. That of the effluent from primary, secondary and tertiary pond of the system is 49.18, 27.9, and 19.8.mg/l respectively. Approximate 88, 93, and 95 % of BOD5 are removed in each pond. The mean yearly SS concentration of influent is 360.5 mg/l That of the effluent from each pond is 53.4, 45.7, and32.7mg/l respectively. Approximate 86, 88, and 91% of SS are removed in each pond. The $BOD_5$ concentration of secondary and tertiary pond can satisfy 30mg/l secondary treatment standard. The SS concentration of effluent from tertiary pond, however, is slightly greater than the standard, which results from activities of carps growing in the pond. The average yearly total nitrogen concentration of influent is 206.8mg/l and that of the effluent from each pond is 48.6, 30.8, and 21.0mg/l respectively. Approximate 74, 88, and 90% of total nitrogen are removed in each pond. The mean yearly total phosphorous concentration of influent is 20.7mg/l and that of the effluent from each pond is 5.3, 3.2, and 2.1mg/l respectively. Approximate 97, 98, and 99% of total phosphorous are removed in each pond. The high removal of nitrogen and phosphorous results from active growth of algae in the upper layer of pond water. Important pond design parameters for southern part of Korea -- areal loading of BOD5, liquid depth, hydraulic detention time, free board, and pond arrangement -- are taken up.

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The Effect of LhGH on Hair Regeneration in C57BL/6CrN Mouse (LhGH가 마우스(C57BL/6CrN)의 모발 재성장에 미치는 영향)

  • Kim, Yong-Ju;Kim, Tae-Keun;Min, Byoung-Hoon;Kim, Soo-Jin
    • Applied Microscopy
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    • v.41 no.1
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    • pp.47-53
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    • 2011
  • Hair is an appendage of skin which protects the body from outer physical and chemical stimuli. Hair is generated from the hair follicle lying on a sunken basal layer of epidermis. Hair cycling, which regenerates hair follicles throughout the life time of the organism. Numerous kinds of factors which exist at the hair follicle have been reported to regulate hair cycling, Human growth hormone secreted from pituitary gland, initially demonstrated to accelerate organ's growth, has been reported to play a role in the biology of organ size determination. We investigated the effect of 6-histidines residues tagged at amino-terminus of human growth hormone using light and electronmicroscopic methods. Human growth hormone encapsulated in nano-liposome (LhGH) was used to find how LhGH affects hair follicle cycling of mouse (C57BL6/CrN). Distilled water as a negative control, 3% Minoxidil as a positive control, and LhGH were applied to mouse for weeks. LhGH increased the number of exposed hairs per given areas ($1mm^2$). This result was also confirmed using a different breed of mice which show natural hair loss in an old age (about 17 months after birth). When LhGH was applied for 3 weeks after natural hair loss, natural hair loss on these mice was prevented, However, the control group mice on which LhGH was not applied showed further hair loss. This result indicates that LhGH may stimulate hair cycling of mouse. In clusion, it is cleat that the LhGH increased the number of hair on mice and help the depilated skin to grow new hair follicles again.