• Title/Summary/Keyword: Protocol Development

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The Effects of the Warm Ischemic Time, the Preserving Temperature and the Cryopreservation Solution on the Viability of Tracheas (온혈허혈시간과 냉동보존온도와 보존액 조성에 따른 기관의 생육성 비교)

  • Sa, Young-Jo;Park, Jae-Kil;Sim, Sung-Bo;Jin, Ung;Moon, Young-Kyu;Lee, Sun-Hee;Jo, Kuhn-Hyun
    • Journal of Chest Surgery
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    • v.42 no.3
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    • pp.283-291
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    • 2009
  • Background: Tracheal reconstruction after extended tracheal resection still remains as a major surgical challenge because good clinical outcomes are usually correlated with limited tracheal resection. Recent investigations with a using cryopreserved trachea for the reconstruction of a trachea have been carried out to overcome this problem. In this study, we analyzed viability of tracheas, which is an important determining factor for the success of transplanting a cryopreserved trachea and the development of post-transplantation tracheal stenosis, according to three different experimental factors: 1) the warm-ischemic time, 2) the cryopreservation solution and 3) the preserving temperature, to determine a better cryopreservation protocol and a better composition of the cryopreservation solution. Material and Method: Rats tracheas were harvested for different warm-ischemic times (0 hr, 12 hrs, 24 hrs). The tracheas were treated with recombinant insulin growth factor-1 (IGF) and they were stored at three different temperatures $(4^{\circ}C,\;-80^{\circ}C,\;-196^{\circ}C)$ for two weeks. After two weeks, we thawed the stored trachea and isolated the cells of the tracheas with using type II collagenase. We cultured the cells for seven days and then we compared the cellular viability by the MTT reduction assay. Result: Though cryopreservation is required to preserve a trachea for a longer time period, the viability of the tracheas stored at $-80^{\circ}C$ and $-196^{\circ}C$ was significantly reduced compared to that of the tracheas stored at $4^{\circ}C$. The viability of the tracheas with warm-ischemic times of 12 hrs and 24 hrs was also reduced in comparison to the tracheas with a warm-ischemic time of 0 hrs. Our data showed that the warm ischemic time and the parameters of crypreservation negatively affect on trachea viability. However, a cryopresrvation solution containing IGF-1 improved the cellular viability better than the existing cryopreservation solution. For the warm ischemic time group of a 0 hr, the addition of IGF-1 improved the viability of trachea at all the preserving temperatures. Conclusion: These experiments demonstrate that the viability of cryopreserved trachea can improved by modifying the components of the crypreservation solution with the addition of IGF-1 and reducing the warm-ischemic time.

Comparison of Cleaning Performance of CFC 113 and the Alternatives (CFC 113과 대체세정제의 세정성능 비교)

  • Row, Kyung Ho;Choi, Dai-Ki;Lee, Youn Yong
    • Analytical Science and Technology
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    • v.6 no.5
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    • pp.521-530
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    • 1993
  • According to the Montreal Protocol, CFC 113, one of the ozone-depleting substances, will be prohibited to use as a cleaning solvent essentially in the electronic industry. Therefore, the development of the alternative cleaning solvents to CFC 113 is being accelerated. A number of the alternative cleaning solvents are avialable on the market. The alternatives of Axarel 32(DuPont), Cleanthru 750H(KAO Chemical), and EC-Ultra(Petroferm) are chosen for the comparison of cleaning performance with CFC 113. The test methods for measuring the cleaning performance were composed of the measurement of the physical properties, the experiments on the material compatibility with cleaning solvents, the measurement of the evaporation rate, and finally the experiments of the removal efficiency. Normally the basic physical properties of the alternatives had higher boiling points, viscosity and surface tension, which were quite different to those of CFC 113. In terms of solubility of rosin-based flux, the solubilities of abietic acid (nonpolar organic) were similar, but those of the activator (polar organic) in the alternatives were better than CFC 113. The evaporation of the alternatives was very slow, compared to CFC 113, which had much lower boiling point. All the cleaning solvents showed the good material compatibility with FR4 and Cu-coated PCB. The better removal efficiencies of abietic acid were obtained when using the ultrasonic mechanical energy over the dipping method. The experiments also indicated the very slow-eavaporating solvent was not desirable with the dipping cleaning method, and the differences in the removal efficiency of the alternatives with the ultrasonic cleaning method were negligible. Among the alternatives, the overall cleaning performances were obsorved as almost similar. Before selecting the ultimate cleaning solvent, the application of cleaning machine, environmental issues, and economics are simultaneously considered with the cleaning performance.

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Report for Development of Korean Portable Cardiopulmonary Bypass II. Experimental Study of Portable Cardiopulmonary Bypass for Emergency Cardiopulmonary Resuscitation after Cardiac Arrest in Normal Dogs (한국형 이동식 심폐소생기 개발 보고 II. 응급소생술을 위한 이동식 심폐소생기의 동물 실험 연구)

  • Kim, Hyoung-Mook;Lee, In-Sung;Baek, Man-Jong;Sun, Kyung;Kim, Kwang-Taik;Lee, Hye-Won;Lee, Kyu-Back;Chang, Jun-Kuen;Kim, Chong-Won;Kim, Hark-Jei
    • Journal of Chest Surgery
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    • v.31 no.12
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    • pp.1147-1158
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    • 1998
  • Background: Portable cardiopulmonary bypass(CPB) technique has been used increasingly as a potent and effective option for emergency cardiopulmonary resuscitation(CPR) because it can maintain more stable hemodynamics and provide better survival than conventional CPR techniques. This study was designed to develop a prototype of Korean portable CPB system and, by applying it to CPR, to discriminate whether it would be superior to standard open-chest CPR. Material and Method: By using adult mongrel dogs, open-chest CPR(OCPR group, n=4) and portable-CPB CPR(CPB group, n=4) were compared with respects to restoration of spontaneous circulation(ROSC), hemodynamics, effects on blood cells, blood gas patterns, biochemical markers, and survivals. Ventricular fibrillation-cardiac arrest(VF-CA) of arrest(VF-CA) of 4 minutes followed by basic life support(BLS) of 15 minutes was applied in either group, which was standardized by the protocol of American Heart Association. Then, advanced life support(ALS) was applied to either group under the support of internal cardiac massage or CPB. ALS was maintained until ROSC was achieved but not longer than 30 minutes regardless of the presence of ROSC. All of the measured values were expressed as means±SD percent change from baseline. Result: During the early ALS, higher mean arterial pressure was maintained in CPB group than in OCPR group(90±19 vs. 71±32 %; p<.05) and lower mean pulmonary arterial pressure was also maintained in CPB group than in OCPR group(105±24 vs. 146±6%; p<.05). ROSC was achieved in all dogs. Post-ROSC levels of hematocrit, RBC, and platelet were decreased and plasma free hemoglobin was increased significantly in CPB group compared to OCPR group(p<.05). Changes in blood gas patterns, lactate, and CK-MB levels were not different between groups. Early mortality was seen in 3 dogs in OCPR group(survival time 31±36 hours) and 2 in CPB group(228±153 hours, p=ns). The remainders in both groups showed prolonged survival. Conclusion: These findings indicate that portable CPB can be effective to maintain stable hemodynamics during cardiac arrest, to achieve ROSC and to prolong survival. Further study is needed to refine the portable CPB system and to meet clinical challenges.

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Strain Improvement through Protoplast Formation and Mutation of Inonotus obliquus Mycelia for Enhanced Production of Innerpolysaccharides (IPS) in Suspended Mycelial Cultures (Inonotus obliquus 의 균사체 액상배양에서 원형질체 형성과 돌연변이를 통한 단백다당체 고생산성 균주 개발)

  • Hong, Hyeong-Pyo;Jeong, Yong-Seob;Chun, Gie-Taek
    • KSBB Journal
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    • v.25 no.2
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    • pp.155-166
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    • 2010
  • Studies on the production of cell-wall bound innerpolysaccharides (IPS) (soluble ${\beta}$-D-glucan) have been performed by use of suspended myelial cultures of Inonotus obliquus. This product has promising potentials as an effective antidiabetic as well as an immunostimulating agents. As a first step to enhanced production of IPS, Intensive strain improvement programs were carried out by obtaining a large amounts of protoplasts for the isolation of single cell colonies. Rapid and large screening of high-yielding producers was possible because about fivefold higher amount of protoplasts ($2.3{\times}10^6$ protoplasts/mL) could be recovered with relatively high regeneration rates of $10^{-2}{\sim}10^{-3}$ by applying a modified filtration method, as compared to the previously used trapping method. A basic protocol necessary for UV-mutation of the protoplasts was also developed, resulting in several overproducing variants with good fermentation properties. Since the amount of IPS extracted from the mycelial cell walls of I. obliquus turned out to be almost constant per g DCW, increase in cell mass was considered the most important factor for the enhancement in IPS production. Therefore, attempts were made to screen mutant cells showing rapid mycelial growth rate in the final suspended cultures. Notably, the mutant strains showing an active cellgrowth in the preceding solid growth cultures were observed to produce higher amount of IPS in the suspended fermentations as well. A striking mutant, OBLQ756-15-5 strain, obtained from the survivors of a harsh UV-treated condition (97% death rate) was found to stably produce as high cell mass as 22 g DCW/L in the final fermentations. Currently, this strain is being tested for development of a scaled-up fermentation process for mass production of IPS.

Effects of Gonadotropin Releasing Hormone on Steroidogenesis and Apoptosis of Human Granulosa-Lutein Cells (생식샘자극호르몬분비호르몬이 사람 과립-황체화 세포의 스테로이드 생성과 세포자연사에 미치는 영향)

  • Lee, Hyo-Jin;Yang, Hyun-Won
    • Development and Reproduction
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    • v.13 no.4
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    • pp.353-362
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    • 2009
  • GnRH and its receptor are known to express locally in the ovary and to regulate the ovarian function by affecting on granulosa and lutein cells. It has been reported that GnRH directly causes apoptosis in the granulosa and lutein cells of the ovary. However, whether the apoptosis of the cells by GnRH is recovered by FSH as an anti-apoptotic factor is not yet known. In this study, we evaluated the apoptosis and the production of progesterone $(P_4)$ and estradiol $(E_2)$ after treatment with 5, 50, and 100 ng/$m\ell$ GnRH and 1 IU/ml FSH in the granulosa-lutein cells that are obtained during oocyte-retrieval for IVF-ET. Results of DNA fragment analysis and TUNEL assay demonstrated that DNA fragmentation and the rate of apoptotic cells were increased in a dose-dependent manner showing a significant increase in the cells treated with 100 ng/$m\ell$ GnRH. In addition, we found that FSH suppresses the apoptosis of the cells induced by GnRH. In the results of chemiluminescence assay for $P_4$ and $E_2$, $P_4$ production was decreased by GnRH treatment, whereas $E_2$ production was not changed. We also demonstrated that FSH inhibits the suppressive effect of GnRH on $P_4$ production as the result of apoptosis. The present results suggest that GnRH agonist using in ovarian hyperstimulation protocol might induce the dysfunction of the ovary, but its function could be recovered by FSH. These results also will be expected to use as the basic data to elucidate the physiological role of GnRH and to develop new ovarian hyperstimulation protocols for IVF-ET.

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Development of soil certified reference material for determination of the hazardous elements (유해원소 측정용 토양 인증표준물질 개발)

  • Kim, In-Jung;Min, Hyung-Sik;Suh, Jung-Ki;Han, Myung-Sub;Lim, Myung-Chul;Kim, Young-Hee;Shin, Sun-Kyoung;Cho, Kyung-Haeng
    • Analytical Science and Technology
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    • v.23 no.5
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    • pp.485-491
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    • 2010
  • A certified reference material (CRM) of KRISS 109-03-SSD was developed for the analysis of hazardous elements in soil. The target elements were As, Cd, Cr, Cu, Hg, Ni, Pb, Zn being regulated by the Soil Environment Conservation Act. Starting material was collected from tailing dump of an unworked tungsten mine at Sangdong (Gangwon-do, Korea). The starting material under-went through a series of fabricating process steps of screening, drying, grinding, sieving, blending, bottling, sterilization and was certified according to the ISO Guide 35. Isotope dilution-inductively coupled mass spectrometry (ID-ICP/MS) and instrumental neutron activation analysis (INAA) were used for the measurement. Homogeneity was tested according to ISO 13528 annex B. The certified values were determined using the results from two different methods or from two independent measurements using a method. Finally, certified values of seven elements of arsenic, cadmium, chromium, copper, lead, nickel and zinc were determined. Mercury did not satisfied the criteria of homogeneity and the result would be provided for information only, together with iron and tungsten. It was also studied, the extractable fraction of elements by aqua regia according to the ISO 11466 protocol being frequently studied for the purpose of environmental monitoring. It was performed as a inter-laboratory study by 6 laboratories of a public institute and universities. Standard deviation among the laboratories was much bigger than the uncertainty of the certified value. The result of inter-laboratory study on the extractable fraction will be provided for information, only.

Sequential Chemoradiotherapy for Stage I/II Nasal Natural Killer/T Cell Lymphoma (I/II 병기 비강 Natural Killer/T Cell 림프종에 대한 순차적 항암화학요법과 방사선치료)

  • Noh Young Joo;Ahn Yong Chan;Kim Won Seog;Ko Young Hyeh
    • Radiation Oncology Journal
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    • v.22 no.3
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    • pp.177-183
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    • 2004
  • Purpose: Authors would report the results of sequential CHOP chemotherapy (cyclophosphamide, adriamycin, vincristine, and prednisone) and involved field radiotherapy (IFRT) for early stage nasal natural killer/T-cell Iymphoma (NKTCL). Materials and Methods: Fourteen among 17 patients, who were registered at the Samsung Medical Center tumor registry with stage I and II nasal NKTCL from March 1995 to December 1999 received this treatment protocol. Three to four cycles of CHOP chemotherapy were given at 3 weeks' interval, which was followed by local IFRT including the known tumor extent and the adjacent draining lymphatics. Results: Favorable responses after chemotherapy (before IFRT) were achievable only in seven patients (5 CR's+2 PR's: 50%), while seven patients showed disease progression. There were six patients with local failures, two with distant relapses, and none with regional lymphatic failure. The actuarial overall survival and progression-free survival at 3 years were 50.0% and 42.9%. All the failures and deaths occurred within 13 months of the treatment start. The factors that correlated with the improved survival were the absence of 'B' symptoms, the favorable response to chemotherapy and overall treatment, and the low risk by international prognostic index on univariate analyses. Conclusion: Compared with the historic treatment results by IFRT either alone or followed by chemotherapy, the current trial failed to demonstrate advantages with respect to the failure pattern and survival. Development of new treatment strategy in combining IFRT and chemotherapy is required for improving outcomes.

The Clinical and Cost Effectiveness of Medical Nutrition Therapy for Patients with Type 2 Diabetes Mellitus (제2형 당뇨병환자에서 임상영앙치료의 임상적 효과와 비용효과 연구)

  • Cho, Youn-Yun;Lee, Moon-Kyu;Jang, Hak-Chul;Rha, Mi-Yong;Kim, Ji-Young;Park, Young-Mi;Sohn, Cheong-Min
    • Journal of Nutrition and Health
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    • v.41 no.2
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    • pp.147-155
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    • 2008
  • Medical nutrition therapy (MNT) is considered a keystone of medical treatment of chronic diseases. However, only few studies have evaluated medical and economical outcome of MNT. The study was performed on the patient with type 2 diabetes mellitus to evaluate the effect of clinical and cost-effective outcomes of MNT. Subjects from two general hospitals were randomly assigned to two different groups; One receiving basic nutritional education (BE) (n = 35), and the other receiving intensive nutritional education (IE) (n = 32) for a 6-month clinical trial. The group which received BE had a single visit with a dietitian, while the other group which received IE had an initial visit with a dietitian addition to two visits during the first 4 weeks of the study periods. Anthropometric parameters, blood components, and dietary intake were measures at the beginning of study period and after 6 month. Cost-effective analysis included direct labor costs, educational materials and medication cost difference during 6 months. After 6 month, subjects from IE group showed significant reduction of body weight (p <0.05) and systolic blood pressure (p <0.05), whereas BE group did not show any significant changes. Result from biochemical indices showed glycated hemoglobin concentration was significantly reduced by 0.7% (p <0.05) only in the IE group. The ratio of energy intake to prescribed energy intake decreased significantly in both groups (p <0.05). Mean time taken for a dietitian to educate the subject was 67.9 ${\pm}$ 9.3 min/person for BE group, while 96.4 ${\pm}$ 12.2 min/person for IE group. Mean number of educational materials was 1.9 ${\pm}$ 0.7/person for BE group and 2.5 ${\pm}$ 0.7/person for IE group. Change in glycated hemoglobin level along the 6 month period of study can be achieved with an investment of \88,510/% by implementing BE and \53,691/% by implementing IE. Considering the net cost-effect of blood glucose control and HbA Ic, IE which provides MNT by dietitian had a cost efficiency advantage than that of BE. According to this study, MNT provided by dietitian had a significant improvements in medical and clinical outcomes compared to that of BE intervention. Therefore, MNT protocol should be performed by systemic intensive nutrition care by dietitian in clinical setting to achieve good therapeutic results of DM with lower cost.

Differentiation and Apoptosis of the Mammalian Embryo and Embryonic Stem Cells(ESC): I. Establishment of Mouse ESC and Induction of Differentiation by Reproductive Hormones (포유동물의 배아 및 기간세포의 분화와 세포사멸 기작: I. 생쥐 배아줄기세포의 확립과 분화유도에 미치는 생식호르몬의 영향)

  • 성지혜;윤현수;이종수;김철근;김문규;윤용달
    • Development and Reproduction
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    • v.6 no.1
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    • pp.55-66
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    • 2002
  • Embryonic stem cells(ES cells) are derived from the inner cell mass(ICM) of blastocysts, which have the potentials to remain undifferentiated, to proliferate indefinitely in vitro, to differentiate into the derivates of three embryonic germ layers. ES cells are an attractive model system for studying the initial developmental decisions and their molecular mechanisms during embryogenesis. Additionally, ES cells of significant interest to those characterizing the various gene functions utilizing transgenic and gene targeting techniques. We investigated the effects of reproductive hormones, gonadotropins(GTH) and steroids on the induction of differentiation and expressions of their receptor genes using the newly established mouse ES cells. We collected the matured blastocysts of inbred mice C57BL/6J after superovulation and co-cultured with mitotically inactivated STO feeder cells. After 5 passages, we confirmed the expression alkaline phosphatase(Alk P) activity and SSEA-1, 3, 4 expressions. The protocol devised for inducing ES differentiation consisted of an aggregation steps, after 5 days as EBs in hormone treatments(FSH, LH, E$_2$, P$_4$, T) that allows complex signaling to occur between the cells and a dissociation step, induced differentiation through attachment culture during 7 days in hormone treatments. Hormone receptors were not increased in dose-dependent manner. All hormone receptors in ES cells treated reproductive hormones were expressed lower than those of undifferentiated ES cell except for LHR expression in E$_2$-treated ES cells group. After hormone induced differentiation, at least some of the cells are not terminally differentiated, as is evident from the expression of Oct-4, a marker of undifferentiated. To assess their differentiation by gene expression, we analyzed the expression of 7 tissue-specific markers from all three germ layers. Most of hormone-treated group increased in the expression of gata-4 and $\alpha$ -fetoprotein, suggesting reproductive hormone allowed or induced differentiation of endoderm.

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A Leg Analysis on the Discharge of Cargo Residue at Sea (화물잔류물의 해양 투입처분(배출) 사안에 대한 법률적 분석)

  • Hong, Gi-Hoon;Park, Chan-Ho
    • Journal of the Korean Society for Marine Environment & Energy
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    • v.9 no.4
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    • pp.193-202
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    • 2006
  • The Consultative Meeting of the Convention on the Prevention of Marine Pollution by Dumping of Wastes and other matter, 1972 (London Convention 1972) has requested to International Maritime Organization (IMO) Marine Environmental Protection Committee to collaborate and help clarify a boundary issue between International Convention for the Prevention of Pollution from Shops, 1973 as modified by the Protocol of 1978 (MARPOL) and the London Convention concerning 'dumping' versus 'discharges' during normal operations of ships in 2004, and subsequently established a Joint London Convention/MEPC Correspondence Group. The Contracting Parties to London Convention expressed their environmental concerns on the broad interpretation of the "cargo-associated wastes" by the States, which could be discharged by ships under MARPOL. Regulatory regimes for the cargo residues appear to vary among states. Some countries require fur ships to discharge their cargo wastes into the port reception facility and IMO also recommends doing so. This paper examines the related current national and international legal texts for the regulation of disposal of wastes from ships in order to analyze the current global concern on the marine pollution associated with waste discharge during operations of ships. In particular, we attempt to evaluate the likely marine environmental consequences arising from the disposal of cargo residue using an hypothetical case for the coal cargo residue among bulk cargos in this paper, since location, magnitude and frequency of the discharge of coal cargo residues into the sea adjacent to Korean Peninsula are not readily available. The cargo residues may be discharged to the sea according to MARPOL 73/78; however, its marine environmental consequences can be significant depending upon the characteristics and amounts of wastes to be discharged. Also the public tolerance of the environmental consequences would be widely different among nations. Multilateral environmental agreements, in general, more strictly apply their rules if there are other options to disposal at sea, i.e. port reception facility in this case. Therefore, port reception facilities for the wastes generated by ships are recommended to be further constructed in major national ports in order to reduce the risk of environmental damages during the operations of ships.

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