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Characterization of SID2 that is required for the production of salicylic acid by using β-GLUCURONIDASE and LUCIFERASE reporter system in Arabidoposis (리포트 시스템을 이용한 살리실산 생합성 유전자 SID2의 발현 해석)

  • Hong, Mi-Ju;Cheong, Mi-Sun;Lee, Ji-Young;Kim, Hun;Jeong, Jae-Cheol;Shen, Mingzhe;Ali, Zahir;Park, Bo-Kyung;Choi, Won-Kyun;Yun, Dae-Jin
    • Journal of Plant Biotechnology
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    • v.35 no.3
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    • pp.169-176
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    • 2008
  • Salicylic acid(SA) is a phytohormone that is related to plant defense mechanism. The SA accumulation is triggered by abiotic and biotic stresses. SA acts as a signal molecular compound mediating systemic acquired resistance and hypersensitive response in plant. Although the role of SA has been studied extensively, an understanding of the SA regulatory mechanism is still lacking in plants. In order to comprehend SA regulatory mechanism, we have been transformed with a SID2 promoter:GUS::LUC fusion construct into siz1-2 mutant and wild plant(Col-0). SIZ1 encodes SUMO E3 ligase and negatively regulates SA accumulation in plants. SID2(SALICYLIC ACID INDUCTION DEFICIENT2) is a crucial enzyme of SA biosynthesis. The Arabidopsis SID2 gene encodes isochorismate synthase(ICS) that controls SA level by conversion of chorismate to isochorismate. We compared the regulation of SID2 in wild-type and siz1-2 transgenic plants that express SID2 promoter:GUS::LUC constructs respectively. The expressions of $\beta$-GLUCURONIDASE and LUCIFERASE were higher in siz 1-2 transgenic plant without any stress treatment. SID2 promoter:GUS::LUC/siz1-2 transgenic plant will be used as a starting material for isolation of siz1-2 suppressor mutants and genes involved in SA-mediated stress signaling pathway.

A COMPARISON OF FRACTURE STRENGTHS OF PORCELAIN-FUSED-TO-TITANIUM CROWN AMONG TITANIUM SURFACE COATING TREATMENTS (타이타늄 표면 코팅 처리에 따른 타이타늄도재관의 파절강도 비교)

  • Kim, Ji-Hye;Park, Sang-Won;Vang, Mong-Sook;Yang, Hong-So;Park, Ha-Ok;Lim, Hyun-Pil;Oh, Gye-Jeong;Kim, Hyun-Seung;Lee, Kwang-Min;Lee, Kyung-Ku
    • The Journal of Korean Academy of Prosthodontics
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    • v.45 no.2
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    • pp.203-215
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    • 2007
  • Statement of problem: Titanium and its alloy, with their excellent bio-compatibility and above average resistance to corrosion, have been widely used in the field of dentistry. However, the excessive oxidization of titanium which occurs during the process of firing on porcelain makes the bonding of titanium and porcelain more difficult than that of the conventional metal-porcelain bonding. To solve this problem related to titanium-porcelain bonding, several methods which modify the surfaces, coat the surfaces of titanium with various pure metals and ceramics, to enable the porcelain adhesive by limiting the diffusion of oxygen and forming the adhesive oxides surfaces, have been investigated. Purpose: The purpose of this study was to know whether the titanium-porcelain bonding strength could be enhanced by treating the titanium surface with gold and TiN followed by fabrication of clinically applicable porcelain-fused-to-titanium crown Material and method: The porcelain-fused-to-titanium crown was fabricated after sandblasting the surface of the casting titanium coping with $Al_2O_3$ and treating the surface with gold and TiN coating followed by condensation and firing of ultra-low fusing porcelain. To compare with porcelain-fused-to-titanium crowns, porcelain-fused-to-gold crowns were fabricated and used as control groups. The bonding strengths of porcelain-fused-to-gold crowns and porcelain-fused-totitanium crowns were set for comparison when the porcelain was fractured on purpose to get the experimental value of fracture strength. Then, the surface were examined by SEM and each fracturing pattern were compared with each other Result:Those results are as follows. 1. The highest value of fracture strength of porcelain-fused-to-titanium crowns was in the order of group with gold coating, group with TiN coating, group with $Al_2O_3$ sandblasting. No statistically significant difference was found among the three (P>.05). 2. The porcelain-fused-to-gold crowns showed the highest value in bonding strength. The bonding strength of crowns porcelain-fused-to-titanium crowns of rest groups showed bonding strength reaching only 85%-94% of that of PFG, though simple comparision seemed unacceptable due to the difference in materials used. 3. The fracturing patterns between metal and porcelain showed mixed type of failure behavior including cohesive failure and adhesive failure as a similar patterns by examination with the naked eye and SEM. But porcelain-fused-to-gold crowns showed high incidence of adhesive failure and porcelain-fused-to-titanium crowns showed high incidence of cohesive failure. Conclusion: Above results proved that when fabricating porcelain-fused-to-titanium crowns, treating casting titanium surface with gold or TiN was able to enhance the bonding strength between titanium and porcelain. Mean value of masticatory force was found to showed clinically acceptable values in porcelain bonding strength in all three groups. However, more experimental studies and evaluations should be done in order to get better porcelain bonding strength and various surface coating methods that can be applied on titanium surface with ease.

Enhancement of the solubility of human tissue inhibitor of matrix metallocroteinase-2 (TIMP-2) in E. coli using a modified in vitro mutagenesis (새로운 유전자 재조합 방법을 이용한 대장균에서의 인간 tissue inhibitor of mtrix metalloproteinase-2 (TIMP-2) 유전자의 가용성 발현)

  • Kim, Jong-Uk;Choi, Dong-Soon;Joo, Hyun;Min, Churl-K.
    • KSBB Journal
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    • v.23 no.3
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    • pp.231-238
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    • 2008
  • The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

Biodegradation Characteristics of Monochlorophenols by Wood Rot Fungi (Monochlorophenol의 목질분해균에 의한 분해 특성)

  • Choi, In-Gyu;Lee, Jae-Won;Choi, Don-Ha
    • Korean Journal of Environmental Agriculture
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    • v.21 no.4
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    • pp.261-268
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    • 2002
  • Biodegradation of monochlorophenols by wood rot fungi such as Daldina concentrica, Trametes versicolor and Pleurotus ostreatus was evaluated by determining their resistance or toxic test and biodegradability. The metabolites of monochlorophenols were also analyzed. Among the three fungi, T. versicolor was the most resistant to 200 ppm of 2-, 3- and 4-chlorophenols, and did not show any inhibitory mycellium growth. But D. concentrica had a little inhibition effect at more than 100 ppm of 3- or 4-chlorophenol. Control cultures of P. ostreatus took even 14 days far the completion of mycellium growth, but the hyphal growth was improved when 2- or 3-chlorophenol were added to the culture. In biodegradation analysis, P. ostreatus showed the highest degradation of 2- or 3-chlorophenol, while T. versicolor was the most effective in 4-chlorophenol. D. concentrica and P. ostreatus slowly degraded 4-chlorophenol. However, T. versicolor had similar degradation capability in the three monochlorophenols, suggesting that the biode- gradation nude is dependent on the fungi as well as the type of monochlorophenol. Several metabolites such as 1,3,5-trihydroxyl benzene, 1-ethyl-1-hydroxyl pentane, 2-propenoicacid, methylmalonic acid and 2-methyl-4-keto-pentan-2-ol were found as products of primary oxidation of 2-, 3- and 4-chlorophenols by intact fungal cultures. fatty acids including tetradecanoic, heptadecanoic and octadecanoic acids were also detected The order of increase of mycellium weight during incubation were P. ostreatus > T. versicolor > D. concentrica. The pH in the culture was not constantly changed depending on incubation days, but the mycellium weight was slightly increased, indicating that the biodegradation of monochlorophenol might have low relationship with the mycellium growth Laccase activities of T. versicolor and P. ostreatus were continuously increased depending on the incubation days, suggesting that the ligninolytic enzyme activity play an important role in the biodegradation of monochlorophenol.

Study of Polysulfone Membrane for Membrane-covered Oxygen Probe System (산소 전극 시스템에 사용되는 polysulfone막에 대한 연구)

  • Hong, Suk In;Kim, Hyun Joon;Park, Hee Young;Kim, Tae Jin;Jeong, Yong Seob
    • Applied Chemistry for Engineering
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    • v.7 no.5
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    • pp.877-887
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    • 1996
  • The ideal membranes for membrane-covered oxygen probes system should be selectively permeable for oxygen and chemically inert, and have good mechanical strength. Polysulfone(PSf) was selected to develop the membrane for membrane-covered oxygen electrodes system. PSf membranes have properties such as good reproducibility, good mechanical strength, chemical inertness, and high heat resistance. PSf membranes were cast from polymer solution on the glass plate at constant temperature, and casting solvents used were tetrahydrofuran(THF), methylene chloride, and N-methyl-2-pyrrolidone(NMP). Tricresyl phosphate(TCP) as plasicizer was added to PSf to increase the softness of membrane. The permeation characteristics were observed for pure oxygen and nitrogen through pure PSf membranes by variable volume method and membrane-covered electrode system. The permeability coefficients of oxygen and nitrogen measured by variable volume method were slightly decreased with increasing of upstream pressure. The permeation properties of PSf membrane using methylene choride as casting solvent were not affected by the PSf amount of polymer solution. The permeability coefficients of oxygen and nitrogen for PSf membrane containing TCP were very slightly lower than those for pure PSf membrane, but ideal separation factors were slightly higher. The flexibility of PSf membrane containing 2wt% TCP was better than that of pure PSf membrane. It was expected that this increase in flexibility would solve the difficulty of fixing the membrane to the cathode. The membrane-covered oxygen probes system was composed of anode, cathode and electrolyte. The type of the anode was Ag/AgCl half-cell, that of cathode was Ag, and the electrolyte was 4N KCl solution. The result of sampled current voltametry for PSf membrane showed the plateu region at -0.3V~-1.0V. The correlation coefficient of oxygen partial pressure versus current for PSf membrane was relatively high, 0.99949. It was concluded that PSf membrane was the good candidate for the membrane-covered oxygen probes system.

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Microtubule-damaging Chemotherapeutic Agent-mediated Mitotic Arrest and Apoptosis Induction in Tumor Cells (미세소관-손상 항암제 처리에 의한 세포주기의 정지 및 에폽토시스 유도)

  • Jun, Do Youn;Kim, Young Ho
    • Journal of Life Science
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    • v.26 no.3
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    • pp.376-386
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    • 2016
  • Apoptosis induction has been proposed as an efficient mechanism by which malignant tumor cells can be removed following chemotherapy. The intrinsic mitochondria-dependent apoptotic pathway is frequently implicated in chemotherapy-induced tumor cell apoptosis. Since DNA-damaging agent (DDA)-induced apoptosis is mainly regulated by the tumor suppressor protein p53, and since more than half of clinical cancers possess inactive p53 mutants, microtubule-damaging agents (MDAs), of which apoptotic effect is mainly exerted via p53-independent routes, can be promising choice for cancer chemotherapy. Recently, we found that the apoptotic signaling pathway induced by MDAs (nocodazole, 17α-estradiol, or 2-methoxyestradiol) commonly proceeded through mitotic spindle defect-mediated prometaphase arrest, prolonged Cdk1 activation, and subsequent phosphorylation of Bcl-2, Mcl-1, and Bim in human acute leukemia Jurkat T cells. These microtubule damage-mediated alterations could render the cellular context susceptible to the onset of mitochondria-dependent apoptosis by triggering Bak activation, Δψm loss, and resultant caspase cascade activation. In contrast, when the MDA-induced Bak activation was inhibited by overexpression of anti-apoptotic Bcl-2 family proteins (Bcl-2 or Bcl-xL), the cells in prometaphase arrest failed to induce apoptosis, and instead underwent mitotic slippage and endoreduplication cycle, leading to formation of populations with 8N and 16N DNA content. These data indicate that cellular apoptogenic mechanism is critical for preventing polyploid formation following MDA treatment. Since the formation of polyploid cells, which are genetically unstable, may cause acquisition of therapy resistance and disease relapse, there is a growing interest in developing new combination chemotherapies to prevent polyploidization in tumors after MDA treatment.

A New High Grain Yielding Forage Rye Cultivar, "Seedgreen" (종자 생산량이 많은 호밀 신품종 "씨드그린")

  • Han, Ouk-Kyu;Hwang, Jong-Jin;Park, Hyung-Ho;Kim, Dea-Wook;Oh, Young-Jin;Park, Tae-Il;Ku, Ja-Hwan;Kwon, Young-Up;Kweon, Soon-Jong;Park, Kwang-Geun
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.2
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    • pp.105-111
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    • 2015
  • "Seedgreen" (Secale cereal L.), a new rye cultivar was developed by National Institute of Crop Science (NICS), RDA in 2013. It was developed from an open pollination from within 10 rye varieties or lines including "Chochun" in 1995. The line "SR95POP-S1-140-9-1-3-7-5-3" was selected for its excellent agronomic appearance, and was placed in yield trials for three years from 2008 to 2010. The new cultivar was designated "Homil50" and was placed in regional yield trials at the five locations around Korea from 2011 to 2013, during which time the name "Seedgreen" was given. This cultivar is an erect plant type and of a long size, with a dark-green leaf color, a yellowish-white colored, medium-diameter culm, and a brown-colored, medium-size grain. The heading and maturation dates of Seedgreen were April 22 and June 16, which were 3 days and 2 days earlier than that of "Gogu", respectively. Seedgreen also showed better winter hardiness and a greater resistance to lodging and wet injury compared to those of the check cultivar. Over three years, the average dry matter yield of Seedgreen was 8.3 ton $ha^{-1}$ (fresh yield = 39.8 ton $ha^{-1}$), which was harvested in late April and was lower than that of the check cultivar Gogu. The seed productivity of Seedgreen was approximately 4 ton $ha^{-1}$, which was 16 % more than that of the check. Seedgreen was higher to than Gogu in term of protein content (10.5% and 9.7%, respectively), total digestible nutrients (TDN) (58.3% and 57%, respectively), and TDN yield $ha^{-1}$ (4.81 ton and 4.77 ton, respectively). This cultivar is recommended as a fall sowing crop in areas where the average daily minimum-mean temperatures are higher than $-12^{\circ}C$ in January, and as a winter crop for whole-crop forage before the planting of rice or green manure around Korea.

"Youhan", New Whole Crop Barley Cultivar of Hooded Spike and Fine Growth Ability in Spring (초기생육이 빠른 삼차망 청보리 신품종 '유한')

  • Park, Tae-Il;Oh, Young-Jin;Park, Hyoung-Ho;Park, Jong-Chul;Cho, Sang-Kyun;Noh, Jae-Hwan;Kim, Dae-Wook;Song, Tae-Hwa;Chae, Hyun-Seok;Jeung, Jae-Hyun;Hong, Ki-Heung;Bae, Jeong-Suk;Huh, Jae-Young;Jang, Yun-Woo;Kim, Kee-Jong;Park, Ki-Hun;Han, Ouk-Kyu
    • Journal of The Korean Society of Grassland and Forage Science
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    • v.35 no.2
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    • pp.81-86
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    • 2015
  • "Youhan" (Hordeum vulgare L.), a new whole-crop barley cultivar, was developed by the breeding team at National Institute of Crop Science, RDA in 2012. Youhan has the growth habit of III, a light green and mid-sized leaf, hooded and lax-type spikes. The cultivar showed 107 cm of culm length, 641 spikes per $m^2$. The heading date of Youhan was May 1, which is one day later than that of the check cultivar "Yuyeon" in upland, and 2 days earlier than that of Yuyeon in paddy field. The maturation time was similar to check cultivar Yuyeon on June 4 in upland and May 31 in paddy field. In terms of winter hardiness and resistance to lodging and disease, Youhan also performed better than the check cultivar. The average forage dry matter (DM) yield in the regional yield trial was approximately 12.6 ton $ha^{-1}$ and 12.0 ton $ha^{-1}$ in upland and paddy field, respectively, which were 6% and 5% higher than that of the check. The yield also showed 7.3% of crude protein, 26.8% of ADF (acid detergent fiber), 47.8% of NDF (neutral detergent fiber), 67.7% of TDN (total digestible nutrients), and a higher grade of silage quality for the whole-crop barley. Fall sowing cropping of Youhan is recommended only in those areas where the average daily minimum-mean temperatures in January are higher than $-8^{\circ}C$, and it should not be cultivated in mountainous areas of Korea.

Analysis of Vasopressin Receptor Type 2(AVPR2) Gene in a Pedigree with Congenital Nehrogenic Diabetes Insipidus : Identification of a Family with R202C Mutation in AVPR2 Gene (신성요붕증 가계에서 바소프레신 V2 수용체(AVPR2) 유전자 분석 : AVPR2 유전자 R202C 돌연변이의 발견)

  • Park June-Dong;Kim Ho-Sung;Kim Hee-Joo;Lee Yoon-Kyung;Kwak Young-Ho;Ha Il-Soo;Cheong Hae-Il;Choi Yong;Park Hye-Won
    • Childhood Kidney Diseases
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    • v.3 no.2
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    • pp.209-216
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    • 1999
  • Purpose : Nephrogenic diabetes insipidus (NDI) is a rare X-linked disorder associated with renal tubule resistance to arginine vasopressin (AVP). The hypothesis that the defect underlying NDI might be a dysfunctional renal AVPR2 has recently been proven by the identification of mutations in the AVPR2 gene in NDT patients. To investigate the association of mutations in th AVPR2 gene with NDI, we analyzed the AVPR2 gene located on the X chromosome. Methods : We have analyzed the AVPR2 gene in a kindred with X-linked NDI. The proband and proband's mother were analyzed by polymerase chain reaction-single strand conformational polymorphism(PCR-SSCP) and DNA sequencing of the AVPR2 gene. We also have used restriction enzyme analysis of genomic PCR product to evaluate the AVPR2 gene. Results : C to T transition at codon 202, predictive of an exchange of tryptophan 202 by cysteine(R202C) in the third extracellular domain was identified. This mutation causes a loss of Hae III site within the gene. Conclusion : We found a R202C missense mutation in the AVPR2 gene causing X-linked NDI, and now direct mutational analysis is available for carrier screening and early diagnosis.

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Successful 20 hours Canine Allograft Preservation with new Solution Containing Triiodothyronine - Development of new lung preservation solution II - (삼요드티로닌을 포함한 폐보존액을 이용한 20시간 폐보존 - 새로운 폐 보존액의 개발 II -)

  • 성숙환;김영태;김주현
    • Journal of Chest Surgery
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    • v.32 no.5
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    • pp.413-421
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    • 1999
  • Background: Ischemia reperfusion injury is known to contribute to the major causes of the early graft failure in lung transplantation. Triiodothyronine (T3) has been suggested to ameliorate ischemia reperfusion injury from both in vivo and in vitro experiments of various organs. Prospecting its beneficial effect for pulmonary allograft preservation, we made a new solution by adding T3 into the extracellular type dextran solution. Material and Method: Twelve adult mongrel dogs underwent left lung allotransplantation. Six donor dogs were flushed with the new solution(Group 1, n=6), and the remaining six were flushed with Euro-Collins solution to serve as controls(Group 2, n=6). Allografts were stored in each preservation solution for 20 hours at 4$^{\circ}C$. Left single lung transplantations were performed. The right pulmonary artery and the right main bronchus were clamped at 15 minutes after the reperfusion and maintained throughout the experiment to evaluate the transplanted left lung function. Result: Arterial carbon dioxide tension was better in group 1 than in group 2 throughout the experiment period and the difference was statistically significant at 2 hours after reperfusion(28.0${\pm}$3.0 mmHg and 53.1${\pm}$17.4 mmHg, p<0.05). The differences of arterial oxygen partial pressure, peak airway pressure and pulmonary vascular resistance showed no statistical significance. The malondialdehyde(MDA) level, measured from tissue obtained at 120 minutes after reperfusion showed no statistically significant difference. The tissue wet/dry ratio of group 1(649${\pm}$27 %) was significantly lower than that of group 2(686${\pm}$71 %, p<0.05). The microscopic examination revealed varying degrees of injury represented mainly by findings such as perivascular neutrophil infiltration, capillary hemorrhage and interstitial congestion. These findings were less severe in group 1 than those in group 2. Conclusion: The new solution demonstrated superior allograft preservation after 20 hour ischemia compared to Euro-Collins solution in canine single left lung transplantation model, these results suggest that T3 might be a promising agent for pulmonary allograft preservation.

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