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Characterization of Mutations in Bruton's Tyrosine Kinase(Btk) Gene from Unrelated 3 X-linked Agammaglobulinemia(XLA) Families in Korea (국내 X-관련성 범저감마글로불린혈증 세가족에 대한 Bruton's Tyrosine Kinase 단백질 발현 및 유전자 변이 분석)

  • Song, Chang-Hwa;Jo, Eun-Kyeong;Park, Jeong-Kyu;Kim, Jung-Soo;Hong, Soo-Jong;Lee, Jae-Ho
    • Clinical and Experimental Pediatrics
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    • v.45 no.3
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    • pp.302-310
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    • 2002
  • Purpose : X-linked agammaglobulinemia(XLA) is an immunodeficiency caused by abnormalities in Bruton's tyrosine kinase(Btk), and is characterized by a deficiency of peripheral blood B cells. We studied cytoplasmic expression of Btk protein and analyzed the Btk gene in peripheral blood mononuclear cells(PBMC) from three XLA families in Korea. Methods : Heparinized venous blood samples were collected from four XLA patients and additional family members in three unrelated XLA families. Mononuclear cells were separated from their blood and the intracellular Btk protein was characterized by a flow cytometry. The mutation analysis was performed using direct sequencing. Results : Cytoplasmic expression of Btk protein in monocytes was not detected in the patients with XLA. We observed a novel deletion and two point mutations within introns(intron 1 and intron 18) resulting in alternative splicings. In XLA family 2, a 980 bp deletion(from intron 9+191 T to intron 10-215 C) including exon 10 was found in patient P2. He was the only sporadic case in this study, because his mother and brother showed a normal Btk expression by flow cytometry. Conclusion : These identified genetic alterations support the molecular heterogeneity of Btk gene in XLA disease. Additionally, by means of flow cytometric analysis, we diagnosed three hypogammaglobulinemia patients as XLA. Advancements in diagnostic methods has facilitated a prompt and definite diagnosis of this disease.

Some New Problems of International Aviation Security- Considerations Forcused on its Legal Aspects (최근국제항공보안대책(最近國際航空保安対策)의 제간제(諸間題) -특히 법적측면(法的測面)을 중심(中心)으로-)

  • Choi, Wan-Sik
    • The Korean Journal of Air & Space Law and Policy
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    • v.5
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    • pp.53-75
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    • 1993
  • This article is concerned with the comment on "Some New Problems of International Aviation Security-Considerations Forcused on its Legal Aspects". Ever since 1970, in addition to the problem of failure to accept the Tokyo, Hague and Montreal Conventions, there has been also the problem of parties to them, failing to comply with their obligations under the respective treaties, in the form especially of nominal penalties or the lack of any effort to prosecute after blank refusals to extradite. There have also been cases of prolonged detention of aircraft, passengers and hostages. In this regard, all three conventions contain identical clauses which submit disputes between two or more contracting States concerning the interpretation or application of the respective conventions to arbitration or failing agreement on the organization of the arbitration, to the International Court of Justice. To the extent to which contracting States have not contracted out of this undertaking, as I fear they are expressly allowed to do, this promision can be used by contracting States to ensure compliance. But to date, this avenue does not appear to have been used. From this point of view, it may be worth mentioning that there appears to be an alarming trend towards the view that the defeat of terrorism is such an overriding imperative that all means of doing so become, in international law, automatically lawful. In addition, in as far as aviation security is concerned, as in fact it has long been suggested, what is required is the "application of the strictest security measures by all concerned."In this regard, mention should be made of Annex 17 to the Chicago Convention on Security-Safeguarding International Civil Aviation against Acts of Unlawful Intereference. ICAO has, moreover, compiled, for restricted distribution, a Security Manual for Safeguarding Civil Aviation Against Acts of Unlawful Interference, which is highly useful. In this regard, it may well be argued that, unless States members of ICAO notify the ICAO Council of their inability to comply with opecific standards in Annex 17 or any of the related Annexes in accordance with Article 38 of the 1944 Chicago Convention on International Civil Aviation, their failure to do so can involve State responsibility and, if damage were to insure, their liability. The same applies to breaches of any other treaty obligation. I hope to demonstrate that although modes of international violence may change, their underlying characteristics remain broadly similar, necessitating not simply the adoption of an adequate body of domestic legislation, firm in its content and fairly administered, but also an international network of communication, of cooperation and of coordination of policies. Afurther legal instrument is now being developed by the Legal Committee of ICAO with respect to unlawful acts at International airports. These instruments, however, are not very effective, because of the absence of universal acceptance and the deficiency I have already pointed out. Therefore, States, airports and international airlines have to concentrate on prevention. If the development of policies is important at the international level, it is equally important in the domestic setting. For example, the recent experiences of France have prompted many changes in the State's legislation and in its policies towards terrorism, with higher penalties for terrorist offences and incentives which encourage accused terrorists to pass informations to the authorities. And our government has to tighten furthermore security measures. Particularly, in the case an unarmed hijacker who boards having no instrument in his possession with which to promote the hoax, a plaintiff-passenger would be hard-pressed to show that the airline was negligent in screening the hijacker prior to boarding. In light of the airline's duty to exercise a high degree of care to provide for the safety of all the passengers on board, an acquiescence to a hijacker's demands on the part of the air carrier could constitute a breach of duty only when it is clearly shown that the carrier's employees knew or plainly should have known that the hijacker was unarmed. The general opinion is that the legal oystem could be sufficient, provided that the political will is there to use and apply it effectively. All agreed that the main responsibility for security has to be borne by the governments. A state that supports aviation terrorism is responsible for violation of International Aviation Law. Generally speaking, terrorism is a violation of international law. It violates the sovereign rights of states, and the human rights of the individuals. We have to contribute more to the creation of a general consensus amongst all states about the need to combat the threat of aviation terrorism. I think that aviation terrorism as becoming an ever more serious issue, has to be solved by internationally agreed and closely co - ordinated measures.

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A Combat Effectiveness Evaluation Algorithm Considering Technical and Human Factors in C4I System (NCW 환경에서 C4I 체계 전투력 상승효과 평가 알고리즘 : 기술 및 인적 요소 고려)

  • Jung, Whan-Sik;Park, Gun-Woo;Lee, Jae-Yeong;Lee, Sang-Hoon
    • Journal of Intelligence and Information Systems
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    • v.16 no.2
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    • pp.55-72
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    • 2010
  • Recently, the battlefield environment has changed from platform-centric warfare(PCW) which focuses on maneuvering forces into network-centric warfare(NCW) which is based on the connectivity of each asset through the warfare information system as information technology increases. In particular, C4I(Command, Control, Communication, Computer and Intelligence) system can be an important factor in achieving NCW. It is generally used to provide direction across distributed forces and status feedback from thoseforces. It can provide the important information, more quickly and in the correct format to the friendly units. And it can achieve the information superiority through SA(Situational Awareness). Most of the advanced countries have been developed and already applied these systems in military operations. Therefore, ROK forces also have been developing C4I systems such as KJCCS(Korea Joint Command Control System). And, ours are increasing the budgets in the establishment of warfare information systems. However, it is difficult to evaluate the C4I effectiveness properly by deficiency of methods. We need to develop a new combat effectiveness evaluation method that is suitable for NCW. Existing evaluation methods lay disproportionate emphasis on technical factors with leaving something to be desired in human factors. Therefore, it is necessary to consider technical and human factors to evaluate combat effectiveness. In this study, we proposed a new Combat Effectiveness evaluation algorithm called E-TechMan(A Combat Effectiveness Evaluation Algorithm Considering Technical and Human Factors in C4I System). This algorithm uses the rule of Newton's second law($F=(m{\Delta}{\upsilon})/{\Delta}t{\Rightarrow}\frac{V{\upsilon}I}{T}{\times}C$). Five factors considered in combat effectiveness evaluation are network power(M), movement velocity(v), information accuracy(I), command and control time(T) and collaboration level(C). Previous researches did not consider the value of the node and arc in evaluating the network power after the C4I system has been established. In addition, collaboration level which could be a major factor in combat effectiveness was not considered. E-TechMan algorithm is applied to JFOS-K(Joint Fire Operating System-Korea) system that can connect KJCCS of Korea armed forces with JADOCS(Joint Automated Deep Operations Coordination System) of U.S. armed forces and achieve sensor to shooter system in real time in JCS(Joint Chiefs of Staff) level. We compared the result of evaluation of Combat Effectiveness by E-TechMan with those by other algorithms(e.g., C2 Theory, Newton's second Law). We can evaluate combat effectiveness more effectively and substantially by E-TechMan algorithm. This study is meaningful because we improved the description level of reality in calculation of combat effectiveness in C4I system. Part 2 will describe the changes of war paradigm and the previous combat effectiveness evaluation methods such as C2 theory while Part 3 will explain E-TechMan algorithm specifically. Part 4 will present the application to JFOS-K and analyze the result with other algorithms. Part 5 is the conclusions provided in the final part.

The Effect of Selenium and Vitamin E on Activity of Enzyme Related to the Lipid Peroxidation in Rat with Alcohol Administration (식이내 Selenium과 Vitamin E가 Alcohol을 섭취한 흰쥐의 간 지질 과산화에 관련된 효소의 활성에 미치는 영향)

  • 김갑순;정승용;김석환
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.22 no.2
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    • pp.116-126
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    • 1993
  • The purposes of this study were to investigate the effect of seleniumc (Se) and vitamin E on activity of enzyme relevant to lipid peroxidation in alcohol administrated rats. Seventy two male rats of Sprague-Dawley strain weighing about 58~62g were divided into 12groups. The dietary Se levels were 0, 0.4 and 10mg and the dietary vitamin E levels were 0 and 150mg per kg diet, respectively. Alcohol-administrated groups received drinking water solution containing 10% of ethanol from the 3-weeks of experimental periods. The obtained experimental results are summarized as follow: The ${\gamma}$-GTP activity in plasma was higher in alcohol administrated groups and high selenium group (HSe) and low selenium group (LSe) than in control groups (CSe). The ${\gamma}$-GOT and GPT activities were higher in alcohol groups. The ${\gamma}$-GTP activity was significantly influenced by alcohol in LSe groups than in other groups. The glutathione peroxidase (GSH-Px) activity of plasma was significantly lower in LSe groups than HSe and CSe groups. The GSH-Px activity of microsomal and cytosolic fraction was slightly lower in alcohol groups and was about a half value lower in HSe and LSe groups than CSe groups. There was negative correlation between plasma Se level and GSH-Px activity of cytosolic fraction in HSe groups (r=- 0.662, p<0.001) and positive correlation in LSe groups (r=0.640, p<0.001). The GSH S-transferase activity in microsomal and cytosolic fraction was slightly higher in alcohol administrated but vitamin E nonadministrated groups, and significantly higher in LSe groups than in other groups. The catalase activity in mitochondria was lower in HSe than CSe groups, but rather higher in LSe groups. The superoxide dismutase (SOD) activity in cytosolic fraction of liver was not found any effect in all groups. The cytochrome P-450 was higher in alcohol groups, but significantly lower in HSe groups. In conclusion, the deficiency of Se and vitamin E develops the hyperoxidation of liver lipid through the increase of activity of enzyme related to the lipid peroxidation and alcohol administration appears to further increase of hyperoxidation of liver lipid.

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『황제내경소문(黃帝內經素問)·칠편대론(七篇大論)』 왕빙 주본(注本)을 통(通)한 운기학설(運氣學說) 관(關)한 연구(硏究)

  • Kim, Gi-Uk;Park, Hyeon-Guk
    • The Journal of Dong Guk Oriental Medicine
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    • v.4
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    • pp.109-140
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    • 1995
  • As we considered in the main subjects, investigations on the theory of 'Doctrine on five elements' motion and six kinds of natural factors(運氣學說)' through 'Wang Bing's Commentary(王氷 注本)' of 'The seven great chapters in The Yellow Emperor's Internal Classic Su Wen' ("黃帝內經素問 七篇大論") are as follows. (1) In The seven great chapters("七篇大論")' Wang Bing supplement theory and in the academic aspects as a interpreter, judging from 'forget(亡)' character. expressed in the 'The missing chapters("素問遺篇")', 'Bonbyung-ron("本病論")' and 'Jabeob-ron(刺法論)', 'The seven great chapters("七篇大論")' must be supplementary work by Wang Bing. Besides, he quoted such forty books as medical books, taoist books, confucianist books, miscellaneous books, etc in the commentary and the contents quoted in the 'Su Wen(素問)' and 'Ling Shu("靈樞")' scripture nearly occupy in the book. As a method of interpreting scripiure as scripture, he edited the order of 'Internal Classic("內經")' ascended from the ancient time and when he compensated for commentary, with exhaustive scholarly mind and by observing the natural phenomena practically and writing the pathology and the methods of treatment. We knew that the book is combined with the study of 'Doctrine on five elements motion and six kinds of natural factors(運氣學說)' (2) When we compare, analyze the similar phrase of 'The seven great chapters in The Yellow Emperor's Internal Classic Su Wen'("黃帝內經素問ㆍ七篇大論") through 'Wang Bing's Commentary(王氷 注本)', he tells abouts organized 'five elements(五行)' and 'heaven's regularly movement(天道運行)' rather than 'Emyangengsangdae-ron("陰陽應象大論")' in 'The seven great chapters("七篇大論")'. Also the 'Ohanunhangdae-ron("五運行大論")' because the repeated sentences with 'Emyangengsangdae-ron("陰陽應象大論")' is long they are omitted. And in the 'Youkmijidae-ron("六微旨大論")', 'Cheonjin ideology(天眞四象)' based on the 'Sanggocheonjin- ron("上古天眞論")', 'Sagijosindae-ron("四氣調神大論")' is written and in the 'Gigoupyondae-ron("氣交變大論")', the syndrome and symptom are explained in detail rather than 'Janggibeobsi-ron("藏氣法時論")', 'Okgijinjang-ron ("玉機眞藏論")' and in the 'Osangieongdae-ron("五常政大論")', the concept of 'five element(五行)' of the 'Gemgwejineon-ron("金櫃眞言論")' is expanded to 'the five elements' motion concept(五運槪念)' and in the 'Youkwonjeonggidae-ron("六元正紀大論")', explanations of 'The five elements' motion and six kinds of natural factors(運氣)' function are mentioned mainly and instead systematic pathology is not revealed rather than 'Emyangengsangdae-ron("陰陽應象大論")'. And in the 'Jijinyodae-ron("至眞要大論")', explanations of the change of atmosphere which correspond to treatment principle by 'The three Yin and Yang(三陰三陽)' as a progressed concepts are revealed. Therefore there are much similarity between the phrase of 'Emyangengsangdae-ron("陰陽應象大論")' and 'chapters of addition(補缺之篇)'. Generally, the doctrine which 'The seven great chapters("七篇大論")' are added by Wang Bing(王氷) is supported because there are more profound concepts rather than the other chapter in 'The seven great chapters("七篇大論")'. (3) When we study Wang Bing's(王氷) 'Pattern on five elements motion and six kinds of natural factors(運氣格局)' in 'The seven great chapter("七篇大論")', in the 'Cheonwongi-dae-ron("天元紀大論")', With 'Cheonjin ideology(天眞思想)' and the concepts of 'Owang(旺)'${\cdot}$'Sang(相)'${\cdot}$'Sa(死)'${\cdot}$'Su(囚)'${\cdot}$'Hu(休)' and 'Cheonbu(天符)'${\cdot}$'Sehwoi(歲會)' are measured time-spacially to the concept of 'Three Sum(三合)' the concept of 'Taeulcheonbu(太乙天符)' is explained. In the 'Ounhangdae-ron("五運行大論")', 'The calender Signs five Sum(天干五合)' is compared to the concepts of 'couples(夫婦)', 'weak-strong(柔强)' and in the 'Youkmijidae-ron("六微旨大論")', 'the relationship of obedience and disobedience(順逆關係)' which conform to the 'energy status(氣位)' change and 'monarch-minister(君相)' position is mentioned. In the 'Gikyobyeondae-ron("氣交變大論")', the concept of 'Sang-duk(相得)', 'Pyungsang(平常)' is emphasized but concrete measurement is mentioned. In the 'Osangieongdae-ron("五常政大論")', the detailed explanation with twenty three 'systemic of the five elements' motion(五運體系)' form and 'rountine-contrary treatment(正治. 反治)' with 'chill-fever-warm-cold(寒${\cdot}$${\cdot}$${\cdot}$凉)' are mentioned according to the 'analyse and differentiate pathological conditions in accordance with the eight principal syndromes(八綱辨證)'. In the 'Youkwonjeonggidae-ron("六元正紀大論")', Wang Bing of doesn't mention the concepts of 'Jungwun(中運)' that is seen in the original classic. In the new corrective edition, as the concepts of 'Jungwun, Dongcheonbu, Dongsehae and Taeulcheonbu(中運, 同天符, 同歲會, 太乙天符)' is appeared, Wang Bing seems to only use the concepts of 'Daewun, Juwun, and Gaekwun(大運, 主運, 客運)'. In the 'Jijinyodaeron("至眞要大論")', Wang Bing added detailed commentary to pathology and treatment doctrine by explaining the numerous appearances of 'Sebo, sufficiency, deficiency(歲步, 有餘, 不足)' and in the relation of 'victory-defeat(勝復)', he argued clearly that it is not mechanical estimation. (4) When we observe the Wang Bing's originality on the study of 'the theory of Doctrine on five elements' motion and six kinds of natural factors(運氣學說)', he emphasized 'The idea of Jeongindogi and Health preserving(全眞導氣${\cdot}$養生思想)' by adding 'Wang Bing's Commentary(王氷 注本)' of 'The seven great chapters("七篇大論")' and explained clearly 'The theory of Doctrine on five elements' motion and six kinds of natural factors(運氣學說)' and simpled and expanded the meaning of 'man, as a microcosm, is connected with the macrocosm(天人相應)' and with 'Atmosphere theory(大氣論)' also explained the meaning of 'rising and falling mechanism(升降氣機)'. In the sentence of 'By examining the pathology, take care of your health(審察病機 無失氣宜)'. he explained the meaning of pathology of 'heart-kidney-water-fire(心腎水火)' and suggested the doctrine and management of prescription. In the estimation and treatment, by suggesting 'asthenia and sthenia(虛實)' two method's estimation, 'contrary treatment(反治)' and treatment principals of 'falling heart fire tonifyng kidney water(降心火益腎水)', 'two class of chill and fever(寒熱二綱)' were demonstrated. There are 'inside and outside in the illness and so inner and outer in the treatment(病有中外 治有表囊)'. This sentence suggests concertedly. 'two class of superfies and interior(表囊二綱)' conforming to the position of disease. Therefore Wang Bing as an excellent theorist and introduced 'Cheoniin ideology(天眞思想)' as a clinician and realized the medical science. With these accomplishes mainly written in 'The theory of Doctrine on five elements' motion and six kinds of natural factors(運氣學說)' of 'The seven great chapters("七篇大論")', he interpreted the ancient medical scriptures and expanded the meaning of scriptures and conclusively contributed to the development of the study 'Korean Oriental Medicine(韓醫學)'.

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Effect of Application Level of Swine Slurry on Agronomic Characteristics and Yield of Corn and $NO_3-N$ Content of Corn Field (돈분액비 시용수준이 사료용 옥수수의 생육특성과 수량 및 토양 중 $NO_3-N$ 함량에 미치는 영향)

  • Lim, Young Chul;Yoon, S. H.;Kim, J. G.;Choi, G. J.;Kim, W. H.;Seo, S.;Lee, S. J.;Yook, W. B.
    • Journal of Animal Environmental Science
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    • v.11 no.2
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    • pp.117-124
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    • 2005
  • Livestock manure has been utilized as fertilizer, and trying to make resources natural circulation of organic material. This experiment was conducted to investigate the effect of application level of swine slurry on agronomic characteristics and yield of silage corn and $NO_3-N$ content of soil for three years in the experimental field of Grassland and Forage Crops Division, National Livestock Research Institute. Summary of the results were as follows. The experiment was conducted according to a randomized complete block design. Six treatments were non fertilizer(NF), chemical fertilizer(CF), $100\%$ swine slurry(SS100), $150\%$ swine slurry(SS150), $200\%$ swine slurry(SS200) and mixture fertilizer(MF, $100\%$ swine slurry + $50\%$ chemical fertilizer) with three replications. The application level of swine slurry(SS) $100\%$ was highest in Brix as $8.6^{\circ}$ and stay green was as swine slurry application increased. Deficiency of plant, plant height, ear height and lodging were increased in above $100\%$ swine slurry application level. Dry matter yield was decreased in SS 100 but increased in SS 150 and SS 200. Significant effect was observed for all treatments. The content of $NO_3-N$ in infiltration water was high in above swine slurry nitrogen $150\%$ but that content in run off water was very small. Conclusively, application to swine slurry below $150\%$ would be recommended to produce higher yield and to conserve environment in corn field.

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Effect of the Application of Zn-contained Granular Fused Phosphate and Compound Fertilizers on the Growth and Yield of Rice in the "Akagare" Paddy Field (적고발생답(赤枯發生畓)에 대(對)한 함아연용성인비(含亞鉛熔成燐肥) 및 복비(複肥)의 시용(施用)이 수도(水稻)의 생육(生育) 및 수량(收量)에 미치는 영향(影響))

  • Cho, S.J.;Cho, T.S.;Yuk, C.S.;Lee, J.Y.
    • Korean Journal of Soil Science and Fertilizer
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    • v.11 no.1
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    • pp.17-24
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    • 1978
  • A field experiment was conducted on a soil where habitual zinc deficiency has been observed, to investigate the effectiveness of two forms of zinc containing fertilizers: zinc enriched fused phosphate and zinc enriched compound fertilizer. The result of present study is summarized as following. 1. The sail used for the study contained relatively large amount of 1N $CH_2COONH_4$ extractable Ca and the pH was 7.03. Available Zn extracted by 0.1 HCl and available $SiO_2$ extracted by NaOAc were 3.35 ppm and 67.7 ppm respectively. 2. In control plots Zinc content of rice plant measured at 20 days after transplanting was 22-23 ppm, which was a little higher than the critical level (20ppm). But at harvesting stage it dropped to 15ppm. 3. The ratios of $P_2O_5/Zn$ and N/Zn tended to lower as the zinc applied to the soil increased. 4. Application of Zinc clearly increased the number of tillers and plant height as compared to the control. It was also observed that the plots received znic headed and matured earlier compared to the control plots by two weeks. 5. Application of zinc increased all of the yield components and the yield of rice. However, there were no statistical differences in yield and yield components among the forms and levels of zinc fertilizers.

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The Study of DNA Mutations of Phenylketonuria in Koreans (한국인에서의 페닐케톤뇨증의 유전자변이에 대한 고찰)

  • Yoo, Su-Jung;Hong, Yong-Hee;Lee, Yong-Wha;Jung, Sung-Chul;Ki, Chang-Seok;Lee, Dong-Hwan
    • Journal of Genetic Medicine
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    • v.5 no.1
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    • pp.26-33
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    • 2008
  • Purpose : Phenylketonuria(PKU) is an inborn error of metabolism and a genetic disorder resulting from a deficiency of phenylalanine hydroxylase(PAH) and decreased activity of tetrahydrobiopterin(BH4).In this study the correlation between the DNA mutation and clinical manifestations was investigated and PAH DNA mutations were compared bewteen Asian and Caucasian populations. Methods : DNA was isolated from peripheral leukocytes. The PAH gene was amplified by Polymerase Chain Reaction(PCR) and the sequence was analyzed with Multiplex Ligation-dependent Probe Amplification(MLPA). Results : We characterized the PAH gene of 102 independent Korean patients with PKU. PAH nucleotide sequence analysis revealed 44 different mutations, including 10 novel mutations comprising 9 missense mutations(N207D, K95del, A447P, G344D, P69S, S391I, A202T, G103S, and I306L) and 1 novel splice-site variant mutation(IVS10-3C>G). R243Q was the most prevalent mutation in this study. A259T has not previously been reported in Asian populations, but we found that this mutation had a frequency of 10.1% in our study. Furthermore, the genotypes of $BH_4$ responsive patients were analyzed and were divided into two groups: $BH_4$ medication-only group and $BH_4$ medication with diet therapy group. In the $BH_4$ medication-only group and $BH_4$ medication with diet therapy group, R241C was the most common mutation. Conclusion : Novel mutations in the PAH gene of PKU patients are still being discovered. Additional information as to the frequency of mutations in the tetrahydrobiopterine responsive gene is also accumulating. We anticipate that knowledge of these PKU gene mutations will assist the diagnosis, genetic counseling, and therapeutic treatment of PKU patients in future.

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Role of Citrate Synthase in Acetate Utilization and Protection from Stress-Induced Apoptosis

  • Lee, Yong-Joo;Kang, Hong-Yong;Maeng, Pil Jae
    • Proceedings of the Microbiological Society of Korea Conference
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    • 2008.05a
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    • pp.39-41
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    • 2008
  • The yeast Saccharomyces cerevisiae has been shown to contain three isoforms of citrate synthase (CS). The mitochondrial CS, Cit1, catalyzes the first reaction of the TCA cycle, i.e., condensation of acetyl-CoA and oxaloacetate to form citrate [1]. The peroxisomal CS, Cit2, participates in the glyoxylate cycle [2]. The third CS is a minor mitochondrial isofunctional enzyme, Cit3, and related to glycerol metabolism. However, the level of its intracellular activity is low and insufficient for metabolic needs of cells [3]. It has been reported that ${\Delta}cit1$ strain is not able to grow with acetate as a sole carbon source on either rich or minimal medium and that it shows a lag in attaining parental growth rates on nonfermentable carbon sources [2, 4, 5]. Cells of ${\Delta}cit2$, on the other hand, have similar growth phenotype as wild-type on various carbon sources. Thus, the biochemical basis of carbon metabolism in the yeast cells with deletion of CIT1 or CIT2 gene has not been clearly addressed yet. In the present study, we focused our efforts on understanding the function of Cit2 in utilizing $C_2$ carbon sources and then found that ${\Delta}cit1$ cells can grow on minimal medium containing $C_2$ carbon sources, such as acetate. We also analyzed that the characteristics of mutant strains defective in each of the genes encoding the enzymes involved in TCA and glyoxylate cycles and membrane carriers for metabolite transport. Our results suggest that citrate produced by peroxisomal CS can be utilized via glyoxylate cycle, and moreover that the glyoxylate cycle by itself functions as a fully competent metabolic pathway for acetate utilization in S. cerevisiae. We also studied the relationship between Cit1 and apoptosis in S. cerevisiae [6]. In multicellular organisms, apoptosis is a highly regulated process of cell death that allows a cell to self-degrade in order for the body to eliminate potentially threatening or undesired cells, and thus is a crucial event for common defense mechanisms and in development [7]. The process of cellular suicide is also present in unicellular organisms such as yeast Saccharomyces cerevisiae [8]. When unicellular organisms are exposed to harsh conditions, apoptosis may serve as a defense mechanism for the preservation of cell populations through the sacrifice of some members of a population to promote the survival of others [9]. Apoptosis in S. cerevisiae shows some typical features of mammalian apoptosis such as flipping of phosphatidylserine, membrane blebbing, chromatin condensation and margination, and DNA cleavage [10]. Yeast cells with ${\Delta}cit1$ deletion showed a temperature-sensitive growth phenotype, and displayed a rapid loss in viability associated with typical apoptotic hallmarks, i.e., ROS accumulation, nuclear fragmentation, DNA breakage, and phosphatidylserine translocation, when exposed to heat stress. Upon long-term cultivation, ${\Delta}cit1$ cells showed increased potentials for both aging-induced apoptosis and adaptive regrowth. Activation of the metacaspase Yca1 was detected during heat- or aging-induced apoptosis in ${\Delta}cit1$ cells, and accordingly, deletion of YCA1 suppressed the apoptotic phenotype caused by ${\Delta}cit1$ mutation. Cells with ${\Delta}cit1$ deletion showed higher tendency toward glutathione (GSH) depletion and subsequent ROS accumulation than the wild-type, which was rescued by exogenous GSH, glutamate, or glutathione disulfide (GSSG). Beside Cit1, other enzymes of TCA cycle and glutamate dehydrogenases (GDHs) were found to be involved in stress-induced apoptosis. Deletion of the genes encoding the TCA cycle enzymes and one of the three GDHs, Gdh3, caused increased sensitivity to heat stress. These results lead us to conclude that GSH deficiency in ${\Delta}cit1$ cells is caused by an insufficient supply of glutamate necessary for biosynthesis of GSH rather than the depletion of reducing power required for reduction of GSSG to GSH.

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The Prognostic Indicies of Pneumocystis Carinii Pneumonia in Immunocompromised Patients other than Acquired Immune Deficiency Syndrome (비 AIDS 면역 결핍 환자들에서 발생한 주폐포자충 폐렴의 예후인자)

  • Park, Wann;Kim, Yoo-Kyum;Lee, Jin-Seong;Ahn, Jong-Jun;Hong, Sang-Bum;Shim, Tae-Sun;Lim, Chae-Man;Lee, Sang-Do;Kim, Woo-Sung;Kim, Dong-Soon;Kim, Won-Dong;Koh, Youn-Suck
    • Tuberculosis and Respiratory Diseases
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    • v.45 no.4
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    • pp.805-812
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    • 1998
  • Background: Among the variety of opportunistic infections, pneumonia comprises the major morbidity in immunocompromised patients. Pneumocystis carnii pneumonia (PCP) and cytomegalovirus (CMV) pneumonia are common infectious illness of immunocompromised hosts. Although there are many reports regarding to the co-infection of PCP and CMV diagnosed by bronchoalveolar lavage (BAL) fluid examination, the effects of CMV co-infection on the outcome of PCP is still controversial. The purpose of this investigation is to evaluate the effects of CMV detected by BAL fluid examination on the clinical course of PCP in the immunocompromised patients other than human immunodeficiency virus infection. Method: Ten patients with PCP were enrolled and retrospective analysis of their medical records were done. HIV infected persons were excluded. The PCP was diagnosed by BAL fluid examination with Calcofluor-White staining. CMV was detected in BAL fluid by Shell-vial culture system. Chest radiographic findings were reviewed. We used Fisher's exact test and Mann-Whitney U test for statistical analysis of data. Results: The underlying disorders of patients were idiopathic pulmonary fibrosis (n=1), renal transplantation (n=4), necrotizing vasculitis (n=l), systemic lupus erythematosus (n=1), brain tumor (n=1), chronic myelogenous leukemia (n=1), unidentified (n=1). There were no difference in clinical course, APACHE III score, arterial blood gas analysis, white blood cell count, lymphocyte count, serum albumin concentration, chest radiographic findings and mortality between patients with PCP alone (n=4) and those with CMV co-infection (n=6). Univariate analysis regarding to the factors that associated with mortality of PCP were revealed that the application of mechanical ventilation (p=0.028), the level of APACHE III score (p=0.018) and serum albumin concentration (p=0.048) were related to the mortality of patients with PCP. Conclusion: The clinical course of PCP patients co-infected by CMV were not different from PCP only patients. Instead, accompanied respiratory failure, high APACHE III score and poor nutritional status were associated with poor outcome of PCP.

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