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Purification and Biological Characterization of Wild-type and Mutants of a Levan Fructotransferase from Microbacterium sp. AL-210 (Microbacterium sp. A-210이 생성하는 Levan fructotransferase의 정제 및 생물학적 특성에 관한 연구)

  • Hwang, Eun-Young;Jeong, Mi-Suk;Cha, Jae-Ho;Jang, Se-Bok
    • Journal of Life Science
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    • v.19 no.9
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    • pp.1218-1225
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    • 2009
  • Difractose anhydrides (DFAs) is studied as a sweetener for diabetics because of its structural property. DFAs have four types: DFA I, III, IV (degradation of levan) and V (degradation of inulin). Especially, DFA IV has been shown to enhance the absorption of calcium in experiments using rats. Levan fructotransferase is an enzyme for producing di-d-fructose-2,6':6,2-dianhydride (DFA IV). To identify structural characterization, we purified wild-type and mutants (D63A, D195N and N85S) of levan fructotransferase (LFTase) from Microbacterium sp. AL-210. These proteins were purified to apparent homogeneity by Ni-NTA affinity column, Q-sepharose ion exchange and gel filtration chromatography and detected by SDS-PAGE. They were also analyzed by circular dichroism (CD) measurements, JNET secondary structure prediction, activity measurements at various temperatures, and pH analysis. The optimum pH for the enzyme-catalyzed reaction was pH 7.5 and optimum temperature was observed at $55^{\circ}C$. Along with wild-type LFTase, mutants were analyzed by CD measurement, fluorescence analysis and differential scanning calorimetry (DSC). N85S showed less $\alpha$-helix and more $\beta$ strand than others. Also, N85S showed almost the same curve as wild-type in their steady-state fluorescence spectra, whereas mutant D63A and D195N showed higher intensity than wild-type. The amino acid sequence of wild-type LFTase was compared to the sequences of exo-inulinase from Aspergillus awamori, a plant fructan 1-exohydrolase from Cichorium intybus, and Thermotogo maritime (Tm) invertase and showed a high identity with Exo-inulinase from Aspergillus awamori.

High Resolution MR Images from 3T Active-Shield Whole-Body MRI System (3T 능동차페형 전신 자기공명영상 장비로부터 얻어진 고해상도 자기공명영상)

  • Bo-Young Choe;Sei-Kwon Kang;Myoung-Ja Chu;Hyun-Man Baik;Euy-Neyng Kim
    • Investigative Magnetic Resonance Imaging
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    • v.5 no.2
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    • pp.138-148
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    • 2001
  • Purpose : Within a clinically acceptable time frame, we obtained the high resolution MR images of the human brain, knee, foot and wrist from 3T whole-body MRI system which was equipped with the world first 37 active shield magnet. Materials and Methods : Spin echo (SE) and Fast Spin Echo (FSE) images were obtained from the human brain, knee, foot and wrist of normal subjects using a homemade birdcage and transverse electromagnetic (TEM) resonators operating in quadrature and tuned to 128 MHz. For acquisition of MR images of knee, foot and wrist, we employed a homemade saddle shaped RF coil. Topical common acquisition parameters were as follows: matrix=$512{\times}512$, field of view (FOV) =20 cm, slice thickness = 3 mm, number of excitations (NEX)=1. For T1-weighted MR images, we used TR = 500 ms, TE = 10 or 17.4 ms. For T2-weighted MR images, we used TR=4000 ms, TE = 108 ms. Results : Signal to noise ratio (SNR) of 3T system was measured 2.7 times greater than that of prevalent 1.5T system. MR images obtained from 3T system revealed numerous small venous structures throughout the image plane and provided reasonable delineation between gray and white matter. Conclusion The present results demonstrate that the MR images from 3T system could provide better diagnostic quali\ulcorner of resolution and sensitivity than those of 1.5T system. The elevated SNR observed in the 3T high field magnetic resonance imaging can be utilized to acquire images with a level of resolution approaching the microscopic structural level under in vivo conditions. These images represent a significant advance in our ability to examine small anatomical features with noninvasive imaging methods.

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Preliminary Study on the MR Temperature Mapping using Center Array-Sequencing Phase Unwrapping Algorithm (Center Array-Sequencing 위상펼침 기법의 MR 온도영상 적용에 관한 기초연구)

  • Tan, Kee Chin;Kim, Tae-Hyung;Chun, Song-I;Han, Yong-Hee;Choi, Ki-Seung;Lee, Kwang-Sig;Jun, Jae-Ryang;Eun, Choong-Ki;Mun, Chi-Woong
    • Investigative Magnetic Resonance Imaging
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    • v.12 no.2
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    • pp.131-141
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    • 2008
  • Purpose : To investigate the feasibility and accuracy of Proton Resonance Frequency (PRF) shift based magnetic resonance (MR) temperature mapping utilizing the self-developed center array-sequencing phase unwrapping (PU) method for non-invasive temperature monitoring. Materials and Methods : The computer simulation was done on the PU algorithm for performance evaluation before further application to MR thermometry. The MR experiments were conducted in two approaches namely PU experiment, and temperature mapping experiment based on the PU technique with all the image postprocessing implemented in MATLAB. A 1.5T MR scanner employing a knee coil with $T2^*$ GRE (Gradient Recalled Echo) pulse sequence were used throughout the experiments. Various subjects such as water phantom, orange, and agarose gel phantom were used for the assessment of the self-developed PU algorithm. The MR temperature mapping experiment was initially attempted on the agarose gel phantom only with the application of a custom-made thermoregulating water pump as the heating source. Heat was generated to the phantom via hot water circulation whilst temperature variation was observed with T-type thermocouple. The PU program was implemented on the reconstructed wrapped phase images prior to map the temperature distribution of subjects. As the temperature change is directly proportional to the phase difference map, the absolute temperature could be estimated from the summation of the computed temperature difference with the measured ambient temperature of subjects. Results : The PU technique successfully recovered and removed the phase wrapping artifacts on MR phase images with various subjects by producing a smooth and continuous phase map thus producing a more reliable temperature map. Conclusion : This work presented a rapid, and robust self-developed center array-sequencing PU algorithm feasible for the application of MR temperature mapping according to the PRF phase shift property.

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Analysis of Image Distortion on Magnetic Resonance Diffusion Weighted Imaging

  • Cho, Ah Rang;Lee, Hae Kag;Yoo, Heung Joon;Park, Cheol-Soo
    • Journal of Magnetics
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    • v.20 no.4
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    • pp.381-386
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    • 2015
  • The purpose of this study is to improve diagnostic efficiency of clinical study by setting up guidelines for more precise examination with a comparative analysis of signal intensity and image distortion depending on the location of X axial of object when performing magnetic resonance diffusion weighted imaging (MR DWI) examination. We arranged the self-produced phantom with a 45 mm of interval from the core of 44 regent bottles that have a 16 mm of external diameter and 55 mm of height, and were placed in 4 rows and 11 columns in an acrylic box. We also filled up water and margarine to portrait the fat. We used 3T Skyra and 18 Channel Body array coil. We also obtained the coronal image with the direction of RL (right to left) by using scan slice thinkness 3 mm, slice gap: 0mm, field of view (FOV): $450{\times}450mm^2$, repetition time (TR): 5000 ms, echo time (TE): 73/118 ms, Matrix: $126{\times}126$, slice number: 15, scan time: 9 min 45sec, number of excitations (NEX): 3, phase encoding as a diffusion-weighted imaging parameter. In order to scan, we set b-value to $0s/mm^2$, $400s/mm^2$, and $1,400s/mm^2$, and obtained T2 fat saturation image. Then we did a comparative analysis on the differences between image distortion and signal intensity depending on the location of X axial based on iso-center of patient's table. We used "Image J" as a comparative analysis programme, and used SPSS v18.0 as a statistic programme. There was not much difference between image distortion and signal intensity on fat and water from T2 fat saturation image. But, the average value depends on the location of X axial was statistically significant (p < 0.05). From DWI image, when b-value was 0 and 400, there was no significant difference up to $2^{nd}$ columns right to left from the core of patient's table, however, there was a decline in signal intensity and image distortion from the $3^{rd}$ columns and they started to decrease rapidly at the $4^{th}$ columns. When b-value was 1,400, there was not much difference between the $1^{st}$ row right to left from the core of patient's table, however, image distortion started to appear from the $2^{nd}$ columns with no change in signal intensity, the signal was getting decreased from the $3^{rd}$ columns, and both signal intensity and image distortion started to get decreased rapidly. At this moment, the reagent bottles from outside out of 11 reagent bottles were not verified from the image, and only 9 reagent bottles were verified. However, it was not possible to verify anything from the $5^{th}$ columns. But, the average value depends on the location of X axial was statistically significant. On T2 FS image, there was a significant decline in image distortion and signal intensity over 180mm from the core of patient's table. On diffusion-weighted image, there was a significant decline in image distortion and signal intensity over 90 mm, and they became unverifiable over 180 mm. Therefore, we should make an image that has a diagnostic value from examinations that are hard to locate patient's position.

Ag Ion Substituted HAp Coatings on Ti-6Al-4V Substrate by IBAD and It's Bactericidal Effect (Ti 합금표면의 항균성 HAp Coating에 관한 연구)

  • Jung, Moon-Young;Kim, Taik-Nam;Kim, Yun-Jong;Yim, Hyuk-Jun;Kim, Jong-Ock;Lim, Dae-young;Kim, Sun-Ok
    • The Journal of Engineering Research
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    • v.3 no.1
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    • pp.189-197
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    • 1998
  • Hyeroxyapatite(HAp) which has good biocompatibility was made by Wet Chemical Process. The surface of Ti-6Al-4V, coated with HAp by lon Beam Assisted Deposition (IBAD), was treated with 5ppm, 10ppm, 20ppm, and 100ppm of $AgNO_3$ solution. In this Ag impregnation process, $Ca^{2+}$ of HAp was substituted with $Ag^+$ of $AgNO_3$. In this study, the antimicrobial effect and biocompatibility of Ti-6Al-4V alloy which was coated with Ag-HAp were examined. The antimicrobial test was carried out with two kinds of bacteria(P. Aeruginosa, S. Epidermidis), which are highly infectious in a transplanting operation of implant materials. As a result of the test, it was observed that Ti-6Al-4V alloy which was treated by 20ppm of $AgNO_3$ solution has good biocompatibility. In order to observe the antimicrobial mechanism of $Ag^+$, E. coli which is the most common bacterium was treated by Ag-HAp. Then cell morphology of E. coli was observed by the transmission electron microscope(TEM). The destruction of cell wall and cytoplasm of E. coil were observed. A black spot appeared in the cytoplasm was analyzed by energy dispersive analysis X-ray (EDAX) and it showed a small amount of $Ag^+$. Thus, it was proved that $Ag^+$ destroys bacteria effectively and Ti-6Al-4V alloy which was impregnate with Ag ion show antimicrobial effect on infection bacteria.

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A Clinical Study of Congenital Intrahepatic Portosystemic Shunt Diagnosed in Neonatal Period (신생아기에 진단된 선천성 간내 문맥 전신성 단락의 임상적 고찰과 치료)

  • Kim, Ji-Young;Kim, Kyung-A;Lee, Yeon-Kyung;Ko, Sun-Young;Shin, Son-Moon;Han, Byung-Hee
    • Neonatal Medicine
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    • v.18 no.1
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    • pp.117-123
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    • 2011
  • Purpose: Congenital intrahepatic portosystemic shunts are rare disease and clinically asymptomatic shunts may be detected by chance on ultrasonogram before and after birth. We studied clinical course, treatment and prognosis of congenital intrahepatic portosystemic shunt at prenatal or neonatal period. Methods: Medical records of 8 patients which were diagnosed in intrahepatic portosystemic shunt in Cheil General Hospital from 2006 through 2010 were reviewed retrospectively. Results: Eight patients with congenital intrahepatic portosystemic shunts were identified. Six patients were diagnosed at prenatal radiological screening, including three cases of intrauterine growth restriction and two cases of preterm baby. One case with increased serum ammonia underwent coil embolization. In four cases including one case that presented elevated direct bilirubin, shunts were closed spontaneously within 11th month after birth. Two patients were diagnosed on abdominal sonogram after birth because of elevated direct hyperbilirubinemia, all of whom presented intrauterine growth restriction. Closure of shunts was confirmed during 4th month to 6th month. Conclusion: Congenital intrahepatic portosystemic shunts are clinically asymptomatic mostly and spontaneous closure is expected within 2 years age. But occasionally they have severe complication, so clinical and radiological observation is needed. Specially in cases of intrauterine growth retardation without evident cause, the possible diagnosis of congenital intrahepatic portosystemic shunts should be considered and prenatal and postnatal examination should be performed. When prenatal diagnosis is made, fetal wellbeing should be monitored periodically until spontaneous closure of shunts.

Heating Performance of Hot Water Supplying System in Greenhouse (온수배관을 이용한 온실의 난방성능)

  • Yoon, Yong-Cheol;Shin, Yik-Soo;Kim, Hyeon-Tae;Bae, Seoung-Beom;Choi, Jin-Sik;Suh, Won-Myung
    • Journal of Bio-Environment Control
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    • v.21 no.2
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    • pp.79-87
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    • 2012
  • This research was conducted to obtain basic data with regard to the heating performance that would be produced by installing an aluminum hot water pipe inside the greenhouse with the goal of reducing the heating energy in greenhouse. The research results are summarized as follows. The degree of difference in relation to the temperature by height within the greenhouse during the entire experiment was significant - within the range of 4.0~$7.0^{\circ}C$. The temperature difference between incoming and outgoing water was about $3.3^{\circ}C$ greater when FCU was activated compared to when it was not activated. Meanwhile, the amount of energy consumed increased about 36.2~40.1%. The amount of pyrexia per hour also increased by 44.6~52.0%. During the experiment period, circulated flux was within the range of 0.48~$0.49L{\cdot}s^{-1}$ while average fluid speed was 1.53~$1.56m{\cdot}s^{-1}$. The average temperature difference between incoming and outgoing water was 6.24~$11.50^{\circ}C$. The amount of heating value by each set temperature within the minimum outdoor temperature range of -14.0~$-0.6^{\circ}C$ was 135,930~307,150 kcal, and the range was within the 9,610~$19,630kcal{\cdot}h^{-1}$ per hour. This demonstrated that about 23~53% heating energy of the maximum heating load could be supplied. Total radiating value and amount of energy consumed were 2,548,306 kcal and 3,075.7 kWh, respectively. When heating takes place using oil, which is a fossil fuel, the total amount of light oil consumed was 281.6 L while the cost was 321,000 won. When the electricity cost for farms is applied, the total cost was about 110,730 won, which is about 33.5% of the cost required compared to oil consumption. The temperature at in the experiment area was about 8.3~$14.6^{\circ}C$ higher compared to that of the control area.

Image Comparisons using a Diethylenetriaminetriacetic Acid Derivative and Gd-DTPA (Diethylenetriaminetriacetic acid 유도체와 Gd-DTPA와의 영상비교)

  • Cho, Jung-Keun;Lee, Chai-Ho;Han, Tae-Jong;Lim, Cheong-Hwan;Kim, Jung-Nam;Choi, Ji-Won
    • The Journal of the Korea Contents Association
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    • v.8 no.11
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    • pp.210-216
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    • 2008
  • In this study, image comparisons were carried out using a MRI contrast medium which was derived by mixing a polyaminocarboxylic ligand and a gadolinium (III) transition metal which is paramagnetic and has good neutron absorbing capabilities with Gd-DTPA which is currently being used widely in the clinical setting. By using a 1.0T (Harmony, SIEMENS) MR equipment, phantoms of which 100cc of saline was diluted with a diethylenetriaminetriacetic acid derivative and Gd-DTPA were imaged. The amount of diethylenetriaminetriacetic acid and Gd-DTPA which was diluted into the 100cc of saline was 0.05mmol/L, 0.1mmol/L, 0.15mmol/L, 0.2mmol/L, 0.3mmol/L, 0.5mmol/L, 1.0mmol/L, 2.0mmol/L, 3.0mmol/L and 4.9mmol/L respectively. Head coils were used and while fixing the SE pulse sequence and image variable (as TE is 14ms, 1NEX with a 256x201 matrix), the signal intensity and simple contrast ratios according to changing concentrations and TR were compared with various TR at 300ms, 400ms, 500ms, 600ms, 700ms, 800ms, 900ms, 1000ms, 1200ms, 1400ms and 1600ms. According to the comparison results of the signal intensity of the image based on changes in contrast medium concentrations and TR, the differences in signal intensity between the two contrast mediums were found to be small at $1.0{\sim}2.0mmol/L$ when the highest signal intensity was achieved. However, at concentrations of 1.0mmol/L or less, the signal intensity was markedly higher in the Diethylenetriaminetriacetic acid derivative than in the Gd-DTPA complex. It was also found that the differences in the signal intensities demonstrated by the concentrations of the contrast mediums were affected by the TR. Accordingly, the efficacy of the Diethylenetriaminetriacetic acid derivative was shown to be better than the Gd-DTPA and also possible to get the optimum image quality by the use of an appropriate TR with appropriate concentrations of contrast medium.

Survival Ability of Porcine Oocytes Frozen-Thawed by Open Pulled Straw Method (Open Pulled Straw 방법에 의해 동결-융해된 돼지난자의 생존능력)

  • 김세웅;박춘근;정희태;양부근;김정익
    • Journal of Embryo Transfer
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    • v.16 no.2
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    • pp.117-125
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    • 2001
  • Vitrification of oocytes has been applied recently fur pigs, but remains elusive. The purpose of this study is to investigate the effects of vitrification in open pulled straws (OPS) on in vitro survival of porcine oocytes. When immature follicular oocytes frozen-thawed were cultured for in vitro maturation, maturation rates to metaphase-II stage were higher in oocytes with (25%) than without (15%) cumulus cells. After In vitro fertilization of oocytes frozen-thawed, the maturation rates were also significantly (P<0.05) higher in oocytes with (41%) that than without (17%) cumulus cells. However, the penetration rates were higher in oocytes without (19%) that than with (9%) cumulus. In another experiment, porcine oocytes matured in vitro were frozen and thawed for in vitro fertilization. The penetration rates were higher than in oocytes without (35%) that than with (26%) cumulus cells. However, the proportions of oocytes dead after in vitro fertilization were significantly (P<0.05) higher in oocytes with that than without cumulus cells. On the other hand, the rates of penetration and dead oocytes at 6 h after in vitro fertilization were not significant differences between oocytes with and without cumulus cells. However, the proportions of dead oocytes with (18%) and without (16%) cumulus cells were higher than in oocytes of control group (0%). These finding indicated the possible broader application for OPS, as they demonstrated that the maturation and fertilization in vitro by frozen-thawed oocytes may be accompained by cumulus cells and culture periods according to the requirements of the survival ability after freezing of mature and immature oocytes in pigs.

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Patients with brain metastases the usefulness of contrast-enhanced FLAIR images after delay (뇌전이 환자의 조영 증강 후 지연 FLAIR 영상의 유용성)

  • Byun, Jae-Hu;Park, Myung-Hwan;Lee, Jin-Wan
    • Korean Journal of Digital Imaging in Medicine
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    • v.16 no.1
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    • pp.13-19
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    • 2014
  • Purpose: FLAIR image is beneficial for the diagnosis of various bran diseases including ischemic CVS, brain tumors and infections. However the border between the legion of brain metastasis and surrounding edema may not be clear. Therefore, this study aims to investigate the practical benefits of delayed imaging by comparing the image from a patient with brain metastasis before a contrast enhancement and the image 10 minutes after a contrast enhancement. Materials and methods: Of the 92 people who underwent MRI brain metastases in suspected patients 13 people in three patients there is no video to target the 37 people confirmed cases, and motion artifacts brain metastases in our hospital June-December 2013, 18 people measurement position except for the three incorrect patient (male: 11 people, female: 7 people, average age: 60 years) in the target, test equipment, 3.0T MR System (ACHIEVA Release, Philips, I was 8ChannelSENSE Head Coil use Best, and the Netherlands). TR 11000 ms, TE 125 ms, TI2800 ms, Slice Thickness 5 mm, gap 5 mm, is a Slice number 21, the parameters of the 3D FFE, T2 FLAIR variable that was used to test, TR 8.1 ms, TE 3.7 ms, Slice number 240 I set to. The experiment was conducted by acquiring the FLAIR prior to contrast enhancement (heretofore referred to as Pre FLAIR), and acquiring the 3D FFE CE five minutes after the contrast enhancement, and recomposing the images in an axial plane of S/T 3mm, G 0mm (heretofore referred to as MPR TRA CE). Using the FLAIR 10 minutes after the contrast enhancement (heretofore referred to as Post FLAIR) and Pi-View, a retrospective study was conducted. Using MRIcro on the image of a patient confirmed for his diagnosis, the images before and after the contrast media, as well as the CNR and SNR of the MPR TRA CE images of the lesion and the site absent of lesion were compared and analyzed using a one-way analysis of variance. Results: CNR for Pre FLAIR and Post FLAIR were 34.35 and 60.13, respectively, with MPR TRA CE at 23.77 showing no significant difference (p<0.050). Post-experiment analysis shows a difference between Pre FLAIR and Post FLAIR in terms of CNR (p<0.050), but no difference in CNR between Post FLAIR and MPR TRA CE (p>0.050), indicating that the contrast media had an effect only on Pre FLAIR and Post FLAIR. The SNR for the normal site Pre FLAIR was 106.43, and for the lesion site 140.79. Post FLAIR for the normal site was 107.79, and for the lesion site 167.91. MPR TRA CE for the normal site was 140.23 and for the lesion site 183.19, showing significant difference (p<0.050), and post-experiment analysis shows that there was a difference in SNR only on the lesion sites for Pre FLAIR and Post FLAIR (p<0.050). There was no difference in SNR between the normal site and lesion site for Post FLAIR and MPR TRA CE, indicating no effect from the contrast media (p>0.050). Conclusions: This experiment shows that Post FLAIR has a higher contrast than Pre FLAIR, and a higher SNR for lesions, It was not not statistically significant and MPR TRA CE but CNR came out high. Inspection of post-contrast which is used in a high magnetic field is frequently used images of 3D T1 but, since the signal of the contrast medium and the blood flow is included, this method can be diagnostic accuracy is reduced, it is believed that when used in combination with Post FLAIR, and that can provide video information added to the diagnosis of brain metastases.

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