• Title/Summary/Keyword: Activation method

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Characterization of a Psychrophilic Metagenome Esterase EM2L8 and Production of a Chiral Intermediate for Hyperlipemia Drug (메타게놈유래의 저온성 에스터라제 EM2L8의 효소적 특성과 이를 활용한 고지혈증 치료제 키랄소재의 생산)

  • Jung, Ji-Hye;Choi, Yun-Hee;Lee, Jung-Hyun;Kim, Hyung-Kwoun
    • Microbiology and Biotechnology Letters
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    • v.37 no.2
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    • pp.118-124
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    • 2009
  • Esterase EM2L8 gene isolated from deep sea sediment was expressed in Escherichia coli BL21 (DE3) and the esterase activity of the cell-free extract was assayed using p-nitrophenyl butyrate-spectrophotometric method. Its optimum temperature was $40-45^{\circ}C$ and 45% activity of the maximum activity was retained at $15^{\circ}C$. The activation energy at $15-45^{\circ}C$ was calculated to be 4.9 kcal/mol showing that esterase EM2L8 was a typical cold-adapted enzyme. Enzyme activity was maintained for 6 h and 4 weeks at $30^{\circ}C$ and $4^{\circ}C$, respectively. When each ethanol, methanol, and acetone was added to the reaction mixture to 15% concentration, enzyme activity was maintained. In the case of DMSO, enzyme activity was kept up to 40% concentration. (S)-4-Chloro-3-hydroxy butyric acid is a chiral intermediate for the synthesis of Atorvastatin, a hyperlipemia drug. When esterase EM2L8 (40 U) was added to buffer solution (1.2 mL, pH 9.0) containing ethyl-(R,S)-4-chloro-3-hydroxybutyrate (38 mM), it was hydrolyzed into 4-chloro-3-hydroxy butyric acid with a rate of $6.8\;{\mu}mole/h$. The enzyme hydrolyzed (S)-substrate more rapidly than (R)-substrate. When conversion yield was 80%, e.e.s value was 40%. When DMSO was added, hydrolysis rate increased to $10.4\;{\mu}mole/h$. The plots of conversion yield vs e.e.s in the presence or absence of DMSO were almost same, implying that the reaction enantioselectivity was not changed by the addition of DMSO. Taken together, esterase EM2L8 had high activity and stability at low temperatures as well as in various organic solvents/aqueous solutions. These properties suggested that it could be used as a biocatalyst in the synthesis of useful pharmaceuticals.

The Lymphocyte Dependent Bactericidal Assay of Human Monocyte and Alveolar Macrophage for Mycobacteria (마이코박테리아에 대한 인체 말초혈액 단핵구와 폐포대식세포의 림프구 의존적 살해능에 관한 연구)

  • Cheon, Seon-Hee;Lee, You-Hyun;Lee, Jong-Soo;Bae, Ki-Sun;Shin, Sue-Yeon
    • Tuberculosis and Respiratory Diseases
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    • v.53 no.1
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    • pp.5-16
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    • 2002
  • Background : Though mononuclear phagocytes serve as the final effectors in killing intracellular Mycobacterium tuberculosis, the bacilli readily survive in the intracellular environment of resting cells. The mechanisms through which cellular activation results in the intracellular killing is unclear. In this study, we sought to explore an in vitro model of a low-level infection of human mononuclear phagocytes with MAC and $H_{37}Ra$ and determine the extent of the lymphocyte dependent cytotoxicity of human monocytes and alveolar macrophages. Materials and Methods : The peripheral monocytes were prepared using the Ficoll gradient method from PPD positive healthy people and tuberculosis patients. The alveolar macrophages were prepared from PPD positive healthy people via a bronchoalveolar lavage. The human mononuclear phagocytes were infected at a low infection rate (bacilli:phagocyte 1:10) with MAC(Mycobacterium avium) and Mycobacterium tuberculosis $H_{37}Ra$. Non-adherent cells(lymphocyte) were added at a 10:1 ratio. After 1,4, and 7 days culture in $37^{\circ}C$, 5% CO2 incubator, the cells were harvested and inoculated in a 7H10/OADC agar plate for the CFU assay. The bacilli were calculated with the CFU/$1{\times}10^6$ of the cells and the cytotoxicity was expressed as the log killing ratio. Results : The intracellular killing of MAC and $H_{37}Ra$ within the monocyte was greater in patients with tuberculosis compared to the PPD positive controls (p<0.05). Intracellular killing of MAC and $H_{37}Ra$ within the alveolar macrophage appeared to be greater than that within the monocytes of the PPD positive controls. There was significant lymphocyte dependent inhibition of intracellular growth of the mycobacteria within the monocytes in both the controls and tuberculosis patients and within the macrophages in the controls(p<0.05). There was no specific difference in the virulence between the MAC and the $H_{37}Ra$. Conclusion : This study is an in vitro model of a low-level infection with MAC and $H_{37}Ra$ of human mononuclear phagocytes. The intracellular cytotoxicity of the mycobacteria within the phagocytic cells was significantly lymphocyte dependent. During the 7 days culture after the intracellular phagocytosis, the actual confinement of the mycobacteria was observed within the monocytes of tuberculosis patients and the alveolar macrophages of the controls as in the case of adding lymphocytes.

The Comparison of Susceptibility Changes in 1.5T and3.0T MRIs due to TE Change in Functional MRI (뇌 기능영상에서의 TE값의 변화에 따른 1.5T와 3.0T MRI의 자화율 변화 비교)

  • Kim, Tae;Choe, Bo-Young;Kim, Euy-Neyng;Suh, Tae-Suk;Lee, Heung-Kyu;Shinn, Kyung-Sub
    • Investigative Magnetic Resonance Imaging
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    • v.3 no.2
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    • pp.154-158
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    • 1999
  • Purpose : The purpose of this study was to find the optimum TE value for enhancing $T_2^{*}$ weighting effect and minimizing the SNR degradation and to compare the BOLD effects according to the changes of TE in 1.5T and 3.0T MRI systems. Materials and Methods : Healthy normal volunteers (eight males and two females with 24-38 years old) participated in this study. Each volunteer was asked to perform a simple finger-tapping task (sequential opposition of thumb to each of the other four fingers) with right hand with a mean frequency of about 2Hz. The stimulus was initially off for 3 images and was then alternatively switched on and off for 2 cycles of 6 images. Images were acquired on the 1.5T and 3.0T MRI with the FLASH (fast low angle shot) pulse sequence (TR : 100ms, FA : $20^{\circ}$, FOV : 230mm) that was used with 26, 36, 46, 56, 66, 76ms of TE times in 1.5T and 16, 26, 36, 46, 56, 66ms of TE in 3.0T MRI system. After the completion of scan, MR images were transferred into a PC and processed with a home-made analysis program based on the correlation coefficient method with the threshold value of 0.45. To search for the optimum TE value in fMRI, the difference between the activation and the rest by the susceptibility change for each TE was used in 1.5T and 3.0T respectively. In addition, the functional $T_2^{*}$ map was calculated to quantify susceptibility change. Results : The calculated optimum TE for fMRI was $61.89{\pm}2.68$ at 1.5T and $47.64{\pm}13.34$ at 3.0T. The maximum percentage of signal intensity change due to the susceptibility effect inactivation region was 3.36% at TE 66ms in 1.5T 10.05% at TE 46ms in 3.0T, respectively. The signal intensity change of 3.0T was about 3 times bigger than of 1.5T. The calculated optimum TE value was consistent with TE values which were obtained from the maximum signal change for each TE. Conclusion : In this study, the 3.0T MRI was clearly more sensitive, about three times bigger than the 1.5T in detecting the susceptibility due to the deoxyhemoglobin level change in the functional MR imaging. So the 3.0T fMRI I ore useful than 1.5T.

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Functional MRI of Visual cortex in the Patients with Occipital Lobe Ischemia (후두엽의 허혈성 뇌졸중 환자에서 시각피질의 기능적 자기공명영상)

  • 이영준;정태섭;윤영수;한승한;조영재;배준호
    • Investigative Magnetic Resonance Imaging
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    • v.3 no.2
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    • pp.173-178
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    • 1999
  • Purpose : To evaluate the usefulness of functional MRI (fMRI) of visual cortex in patients with ischemic infarction in the occipital lobe. Materials and Methods : Four patients with the symptoms and signs of visual cortical ischemia were included. Functional MRI was performed by 2D-FLASH technique with the parameter of 90/56msec TR/TE, $40^{\circ}$ flip angle, $240{\times}240{\;}FOV,{\;}64{\times}128$ matrix number, 8.32 seconds acquisition time, 8mm slice thickness. An axial slice including both visual cortices was selected and alternative activation and resting of the visual cortex was performed using red color photostimulator. all patients undertook visual field test, and vascular abnormality was examined by MRA (n=4) and DSA (n=2). fMRI results were compared with the results of a visual field test, conventional MRI and cerebral angiography. Results : On fMRI, decreased activity of the visual cortex was found in the occipital lobe corresponding to stenosis of the posterior cerebral artery or its branch noted on angiogram. However, 2 of 4 patients showed no abnormal findings on conventional MRI. Visual field defect was noted in 3 patients, one and of whom showed no abnormality on conventional MRI and diffusion-weighted image, but revealed decreased activity in the corresponding visual cortex on fMRI. Conclusion : fMRI may be a sensitive method for detection of the status of decreased blood flow or vascular reserve which other methods can not.

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Extracellular $K^+$ Effects on the Mouse Aortic Endothelial Cell Contractility (쥐 대동맥 혈관 내피세포에서 세포 외 $K^+$에 의한 혈관 수축선 조절 기전)

  • 안재호;유지영
    • Journal of Chest Surgery
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    • v.36 no.12
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    • pp.887-893
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    • 2003
  • External stimuli increases intracellular (IC) $Ca^{2+}$, which increases extracellular (EC) $K^{+}$. To verify $K^{+}$ effects on the vascular contraction, we performed an experiment using mouse aortic endothelial cell. Meterial and Method: We examined the mouse aortic contractility changes as we measured the IC $Ca^{2+}$ change and ionic current by using the voltage clamp technique under different conditions such as: increasing EC $K^{+}$, removing endothelial cell, giving L-NAME (N-nitro-L-arginine methyl ester) which suppress nitric oxide formation, Ouabain which control N $a^{+}$ - $K^{+}$ pump and N $i^{2+}$ which repress N $a^{+}$-C $a^{2+}$ exchanger Result: When we increased EC $K^{+}$ from 6 to 12 mM, there was no change in aortic contractility. Aorta contracted with more than 12 mM of EC $K^{+}$. Ace-tylcholine (ACh) induced relaxation was inhibited with EC $K^{+}$ from 6 to 12 mM, but was not found after de-endothelialization or L-NAME treatment. ATP or ACh increased IC $Ca^{2+}$ in cultured endothelium. After maximal increase of IC $Ca^{2+}$, increasing EC $K^{+}$ from 6 to 12 mM made IC $Ca^{2+}$ decrease and re-decreasing EC $K^{+}$ to 6 mM made IC $Ca^{2+}$ increase. Ouabain and N $i^{2+}$ masked the inhibitory effect of endothelium dependent relaxation by increased EC $K^{+}$. Conclusion: These data indicate that increase in EC $K^{+}$ relaxes vascular smooth muscle and reduces $Ca^{2+}$ in the endothelial cells which inhibit endothelium dependent relaxation. This inhibitory mechanism may be due to the activation of N $a^{+}$- $K^{+}$ pump and N $a^{+}$-C $a^{2+}$ exchanger. $a^{+}$-C $a^{2+}$ exchanger.r.

Analysis of Na and Cl Contents in Children’s Favorite Foods (어린이 선호 간식의 Na와 Cl 함량 분석)

  • Lee, Ok-Hee;Chung, Yong-Sam;Moon, Jong-Wha
    • Journal of Nutrition and Health
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    • v.43 no.5
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    • pp.524-532
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    • 2010
  • The Na, component of salt, can increase the risk of high blood pressure and hypertension. Especially, children are exposed to an increased risk of high sodium intake, because they often consume Na-rich processed foods and commercially prepared foods in the street. However, the database for the sodium and chloride content for these children's favorite foods is insufficient. In this study, the Na and Cl contents in 89 children's favorite foods were analyzed by using Instrumental Neutron Activation Analysis (INAA) method. The analyzed food items were presented after being classified into 33 kinds of food groups. The Na contents in 100 g children's favorite foods ranged from 0.3 mg to 35.1mg in fruits, 28.9mg to 82.5mg in milks, 127.2 mg to 602.2 mg in breads, cakes, sandwiches, and rice cakes, 2.5 mg to 1169.9 mg in candies, cookies and ice creams, 226.9 mg to 693.7 mg in commercially prepared street foods, and 103.4 mg to 875.8 mg in fast foods of westernized restaurant. Among children's favorite food groups, 100 g fried chicken, hotdog, burgers, and donuts contained an average Na of 536 mg, 553 mg, 794 mg, and 562.2 mg, respectively, so they are classified as 'high Na foods'. In contrast, 100 g fruits and dairy products contained Na an average 4.9 mg and 43.4 mg, respectively, being classified as 'low Na foods'. One serving of ramen, mandu noodle, and burger pizza can supply over 667mg Na, which is one third of the KDRI targeted intake. Likewise, the Cl contents in children's favorite foods were diverse according to food groups. The Cl contents in the analyzed foods differed from the 1.5 times of Na content, which is assumed in general. This study showed that the Na and Cl contents in children's favorite foods are very diverse. Conclusively, a strategy to reduce Na contents in the fast foods such as noodles and westernized restaurant foods is necessary for children health.

Hydro-Mechanical Modelling of Fault Slip Induced by Water Injection: DECOVALEX-2019 TASK B (Step 1) (유체 주입에 의한 단층의 수리역학적 거동 해석: 국제공동연구 DECOVALEX-2019 Task B 연구 현황(Step 1))

  • Park, Jung-Wook;Park, Eui-Seob;Kim, Taehyun;Lee, Changsoo;Lee, Jaewon
    • Tunnel and Underground Space
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    • v.28 no.5
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    • pp.400-425
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    • 2018
  • This study presents the research results and current status of the DECOVALEX-2019 project Task B. Task B named 'Fault slip modelling' is aiming at developing a numerical method to simulate the coupled hydro-mechanical behavior of fault, including slip or reactivation, induced by water injection. The first research step of Task B is a benchmark simulation which is designed for the modelling teams to familiarize themselves with the problem and to set up their own codes to reproduce the hydro-mechanical coupling between the fault hydraulic transmissivity and the mechanically-induced displacement. We reproduced the coupled hydro-mechanical process of fault slip using TOUGH-FLAC simulator. The fluid flow along a fault was modelled with solid elements and governed by Darcy's law with the cubic law in TOUGH2, whereas the mechanical behavior of a single fault was represented by creating interface elements between two separating rock blocks in FLAC3D. A methodology to formulate the hydro-mechanical coupling relations of two different hydraulic aperture models and link the solid element of TOUGH2 and the interface element of FLAC3D was suggested. In addition, we developed a coupling module to update the changes in geometric features (mesh) and hydrological properties of fault caused by water injection at every calculation step for TOUGH-FLAC simulator. Then, the transient responses of the fault, including elastic deformation, reactivation, progressive evolutions of pathway, pressure distribution and water injection rate, to stepwise pressurization were examined during the simulations. The results of the simulations suggest that the developed model can provide a reasonable prediction of the hydro-mechanical behavior related to fault reactivation. The numerical model will be enhanced by continuing collaboration and interaction with other research teams of DECOLVAEX-2019 Task B and validated using the field data from fault activation experiments in a further study.

Review of the study on the surfactant-induced foliar uptake of pesticide (계면활성제에 의해 유도되는 농약의 엽면 침투성 연구 현황)

  • Yu, Ju-Hyun;Cho, Kwang-Yun;Kim, Jeong-Han
    • The Korean Journal of Pesticide Science
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    • v.6 no.1
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    • pp.16-24
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    • 2002
  • Research trends in the measurement of foliar uptake of pesticides and the recently proposed action mechanism of the surfactant-induced uptake of pesticides were reviewed with the related reports and studies. Major techniques used in those fields are bioassay, radiotracer techniques with leaves or cuticular membrane. Recently, a new method using Congo Red as a tracer was proposed. The limiting factor in the pesticides uptake into leaves is the waxy layer which consists of the epicuticular and cuticular wax. Physico-chemical parameters such as molar volume, water solubility and partition coefficient of pesticides have limited influences on the pesticide uptake into leaves. Polydisperse ethoxylated fatty alcohol surfactants are well known as the good activator for many pesticides. It is now generally agreed that uptake activation is not related to the intrinsic surface active properties of surfactants such as surface activity, solvent property, humectancy and critical micelle concentration. Recent studies using ESR-spectroscopy revealed that the surfactants have an unspecific plasticising effect on the molecular structure of the wax and cuticular matrix, leading to increased mobilities of pesticides. Penetration of surfactants into waxy layer altered the pesticide mobility in wax and the partition coefficient of pesticide, and then the pesticides penetration into leaves was enhanced temporally. The enhancing effect of surfactant could be significantly different depending on the carbon number of aliphatic moiety and the number of ethoxy group in polyoxyethylene chain of surfactants. It is suggested that the rate of penetration of surfactants should have a significant relationship with the rate of penetration of pesticides.

Kinetics of the Reaction of Carbon Dioxide with AMP and Piperazine (AMP에 Piperazine을 첨가한 CO2 흡수 동역학)

  • Jang, Sang-Yong;Song, Ju-Seouk;Cho, Sang-Won;Oh, Kwang-Joong
    • Journal of Korean Society of Environmental Engineers
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    • v.22 no.3
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    • pp.485-494
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    • 2000
  • According to the worldwide interest in controlling $CO_2$ which contributes to green house effect. new techniques of reducing $CO_2$ are under development. We have developed new technique for reducing $CO_2$. In low $CO_2$ concentration. the chemical absorption method is useful. In this study. the kinetics of the reaction between $CO_2$ and the sterically hindered amine solution with piperazine. have been investigated from measurements of the rate of absorption of $CO_2$ in the stirred vessel that has a horizontal liquid-gas interface, in the temperature range $30{\sim}70^{\circ}C$. The experiments were carried out in the range 10.130~20.260 kPa of partial pressure of $CO_2$, and in aqueous $2.0kmol/m^3$ AMP solution with $0{\sim}0.4kmol/m^3$ piperazine The experimental results are as follows: 1) The absorption rate of $CO_2$ into aqueous AMP + piperazine solution is gett ng faster than that of aqueous AMP absorbents with temperature. Because the activation energy of piperazine 57.147 kJ/mol is higher than that of AMP 41.7kJ/mol. therefore the effect of piperazine on absorption rate increases with temperature. 2) Compared with aqueous AMP solution. the absorption rate of $CO_2$ into AMP + piperazine solution increases from 6.33% at $30^{\circ}C$ to 12% at $70^{\circ}C$, so AMP + piperazine solution is thought to be a better than AMP solution, 3) The reaction rate constants of piprazine in aqueous AMP solution with $CO_2$ have been determined as 217.21, 420.46, 707.00 and $3162.167m^3/kmol{\cdot}s$ respectively at 30, 40, 50 and $70^{\circ}C$ but these results are higher than those of Xu which were 186.7. 367.32. 693.01. $2207.65m^3/kmol{\cdot}s$ at 30, 40, 55, $70^{\circ}C$in aqueous MDEA solution. So the regression analysis of the data has led to the following equation In $k_p$ =28.324-6934.7/T.

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Characterization and annealing effect of tantalum oxide thin film by thermal chemical (열CVD방법으로 증착시킨 탄탈륨 산화박막의 특성평가와 열처리 효과)

  • Nam, Gap-Jin;Park, Sang-Gyu;Lee, Yeong-Baek;Hong, Jae-Hwa
    • Korean Journal of Materials Research
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    • v.5 no.1
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    • pp.42-54
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    • 1995
  • $Ta_2O_5$ thin film IS a promising material for the high dielectrics of ULSI DRAM. In this study, $Ta_2O_5$ thin film was grown on p-type( 100) Si wafer by thermal metal organic chemical vapo deposition ( MCCVD) method and the effect of operating varialbles including substrate temperature( $T_s$), bubbler temperature( $T_ \sigma$), reactor pressure( P ) was investigated in detail. $Ta_2O_5$ thin film were analyzed by SEM, XRD, XPS, FT-IR, AES, TEM and AFM. In addition, the effect of various anneal methods was examined and compared. Anneal methods were furnace annealing( FA) and rapid thermal annealing( RTA) in $N_{2}$ or $O_{2}$ ambients. Growth rate was evidently classified into two different regimes. : (1) surface reaction rate-limited reglme in the range of $T_s$=300 ~ $400 ^{\circ}C$ and (2: mass transport-limited regime in the range of $T_s$=400 ~ $450^{\circ}C$.It was found that the effective activation energies were 18.46kcal/mol and 1.9kcal/mol, respectively. As the bubbler temperature increases, the growth rate became maximum at $T_ \sigma$=$140^{\circ}C$. With increasing pressure, the growth rate became maximum at P=3torr but the refractive index which is close to the bulk value of 2.1 was obtained in the range of 0.1 ~ 1 torr. Good step coverage of 85. 71% was obtained at $T_s$=$400 ^{\circ}C$ and sticking coefficient was 0.06 by comparison with Monte Carlo simulation result. From the results of AES, FT-IR and E M , the degree of SiO, formation at the interface between Si and TazO, was larger in the order of FA-$O_{2}$ > RTA-$O_{2}$, FA-$N_{2}$ > RTA-$N_{2}$. However, the $N_{2}$ ambient annealing resulted in more severe Weficiency in the $Ta_2O_5$ thin film than the TEX>$O_{2}$ ambient.

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