• Title/Summary/Keyword: 연쇄반응

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Fragment Analysis for Detection of the FLT3-Internal Tandem Duplication: Comparison with Conventional PCR and Sanger Sequencing (FLT3-ITD 검출을 위한 절편분석법: 일반 중합효소연쇄반응 및 직접염기서열분석법과의 비교)

  • Lee, GunDong;Kim, Jeongeun;Lee, SangYoon;Jang, Woori;Park, Joonhong;Chae, Hyojin;Kim, Myungshin;Kim, Yonggoo
    • Laboratory Medicine Online
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    • v.7 no.1
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    • pp.13-19
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    • 2017
  • Background: We evaluated a sensitive and quantitative method utilizing fragment analysis of the fms-like tyrosine kinase 3-internal tandem duplication (FLT3-ITD), simultaneously measuring mutant allele burden and length, and verified the analytical performance. Methods: The number and allelic burden of FLT3-ITD mutations was determined by fragment analysis. Serial mixtures of mutant and wild-type plasmid DNA were used to calculate the limit of detection of fragment analysis, conventional PCR, and Sanger sequencing. Specificity was evaluated using DNA samples derived from 50 normal donors. Results of fragment analysis were compared to those of conventional PCR, using 481 AML specimens. Results: Defined mixtures were consistently and accurately identified by fragment analysis at a 5% relative concentration of mutant to wild-type, and at 10% and 20% ratios by conventional PCR and direct sequencing, respectively. No false positivity was identified. Among 481 AML specimens, 40.1% (193/481) had FLT3-ITD mutations. The mutant allele burden (1.7-94.1%; median, 28.2%) and repeated length of the mutation (14-153 bp; median, 49 bp) were variable. The concordance rate between fragment analysis and conventional PCR was 97.7% (470/481). Fragment analysis was more sensitive than conventional PCR and detected 11 additional cases: seven had mutations below 10%, three cases represented conventional PCR failure, and one case showed false negativity because of short ITD length (14 bp). Conclusions: The new fragment analysis method proved to be sensitive and reliable for the detection and monitoring of FLT3-ITD in patients with AML. This could be used to simultaneously assess ITD mutant allele burden and length.

Epidemiology and Clinical Characteristics of Parainfluenza Virus Type 4 in Korean Children: a Single Center Study, 2015-2017 (소아에서 파라인플루엔자 바이러스 4형의 역학 및 임상 양상에 대한 단일기관 연구: 2015-2017)

  • Sohn, Young Joo;Choi, Youn Young;Yun, Ki Wook;Choi, Eun Hwa;Lee, Hoan Jong
    • Pediatric Infection and Vaccine
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    • v.25 no.3
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    • pp.156-164
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    • 2018
  • Purpose: We aimed to identify the epidemiology and the clinical characteristics of human parainfluenza virus type 4 (HPIV-4) infection compared to HPIVs 1-3 infections in Korean children. Methods: We reviewed medical records of children with HPIV infection who visited Seoul National University Children's Hospital from 2015 to 2017. Detection of respiratory viruses was performed using real time-polymerase chain reaction (rt-PCR), which could differentiate HPIVs 1-4. Diagnosis was classified as a febrile illness, upper respiratory tract infection (URI), croup, bronchiolitis, or pneumonia. The epidemiology, demographic features, and clinical characteristics among HPIV types were compared. The clinical data were analyzed only for the previously healthy children. Results: Of the 472 children diagnosed with HPIV infection, 108 (22.9%) were previously healthy: 24 (22.2%), 19 (17.6%), 39 (36.1%), and 26 (24.1%) in HPIV types 1, 2, 3, and 4, respectively. The median age of children with HPIV-4 infection was 11 (0-195) months: the proportion of children aged < 2 years and 2 to < 5 years were 65.4% and 19.2%, respectively. Clinical diagnoses of HPIV-4 infection were bronchiolitis (38.5%), pneumonia (30.8%), and URI (30.8%). Croup was the most prevalent in HPIV-2 (21.1%) and none in HPIV-4 infection (P=0.026). Hospital admission rates among HPIV types were not significantly different (P>0.05). Conclusions: We observed seasonal peak of HPIV-4 infection in 2015 and 2017. HPIV-4 was a common respiratory pathogen causing lower respiratory tract infection in hospitalized children.

Single or Dual Infection with Respiratory Syncytial Virus and Human Rhinovirus: Epidemiology and Clinical Characteristics in Hospitalized Children in a Rural Area of South Korea (호흡기세포융합바이러스와 라이노바이러스의 단독 혹은 동시감염의 역학 및 임상적 특성: 강원 지역 단일 기관의 후향적 연구)

  • Kwon, Yerim;Cho, Won Je;Kim, Hwang Min;Lee, Jeongmin
    • Pediatric Infection and Vaccine
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    • v.26 no.2
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    • pp.99-111
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    • 2019
  • Purpose: Respiratory syncytial virus (RSV) and human rhinovirus (hRV) are the most common causes of child respiratory viral infections. We aimed to investigate epidemiological and clinical characteristics of RSV and hRV single infections and coinfections. Methods: Nasopharyngeal aspirates of hospitalized children aged <5 years were tested using multiplex reverse transcription polymerase chain reaction (RT-PCR) from October 2014 to April 2017. Their medical records were retrospectively reviewed. Results: RSV or hRV was detected in 384 patients who divided into 3 groups: patients with RSV (R group, n=258); patients with hRV (H group, n=99); and patients with both (RH group, n=27). The R group (median age, 6 months) consisted of 248 (96.1%) patients with lower respiratory tract infection (LRTI), and 14 (5.4%) needed oxygen inhalation. Infants aged <12 months (63.2%) had respiratory difficulty and were supplied oxygen more often. The H group (median age, 16 months) consisted of 56 (56.6%) patients with LRTI, 4 (4%) required oxygen inhalation, and 1 (1.0%) required mechanical ventilation. Infants (40.4%) showed longer hospitalization compared to patients aged ${\geq}12$ months (5 vs. 4 days, P<0.05). The RH group consisted of 24 (88.9%) patients with LRTI, and 2 (7.4%) needed oxygen inhalation. Hospitalization days and oxygen inhalation and mechanical ventilation rates did not differ between single infections (R and H groups) and coinfections (RH group). Conclusions: RSV was detected more often in younger patients and showed higher LRTI rates compared to hRV. Single infections and coinfections of RSV and hRV showed no difference in severity.

Efficient virus elimination for apple dwarfing rootstock M.9 and M.26 via thermotherapy, ribavirin and apical meristem culture (사과 왜성대목 M.9 및 M.26의 고온, ribavirin, 생장점 배양을 통한 바이러스 제거)

  • Kwon, Young Hee;Lee, Joung Kwan;Kim, Hee Kyu;Kim, Kyung Ok;Park, Jae Seong;Huh, Yoon Sun;Park, Eui Kwang;Yoon, Yeo Joong
    • Journal of Plant Biotechnology
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    • v.46 no.3
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    • pp.228-235
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    • 2019
  • Apple (Malus pumila) is one of the most economically important fruits in Korea. but virus infection has decreased the sustainable production of apples and caused serious problems such as yield loss and poor fruit quality. Virus or viroid infection including apple chlorotic leaf spot virus (ACLSV), apple stem pitting virus (ASPV), apple stem grooving virus (ASGV), apple mosaic virus (ApMV) and apple scar skin viroid (ASSVd) have been also reported in Korea. In many cases, as apple gets infected with virus and viroid with no specific symptoms, the damage and symptoms caused by the viruses are not detected. In our research, viruses in the rootstock were eliminated for a virus-free apple dwarfing rootstock of M.9 and M.26. The virus elimination methods were apical meristem culture, thermotherapy ($37^{\circ}C$, 6 weeks) and chemotherapy($Ribavirin^{(R)}$). The detection of apple viruses was accomplished by Enzyme-linked Immuno-Sorbent Assay (ELlSA) and reverse transcription-polymerase chain reaction (RT-PCR). RT- PCR method was 10 ~ 30% more sensitive than the ELISA method. The efficiency of virus elimination was enhanced in apical meristem culture method. The acquisition rate of virus-free apple dwarfing rootstocks was 30 ~ 40% higher in apical meristem culture. After the meristem culturing of M.9, the infection ratio of ACLSV, ASPV and ASGV was 45%, 60% and 50%, respectively. In the apple dwarfing rootstock of M.26, the infection ratio of ACLSV, ASPV and ASGV was 40%, 55% and 55%, respectively. Based on this study, the best method for the production of virus-free apple dwarfing rootstocks was the apical meristem culture.

Resveratrol Ameliorates NMDA-induced Mitochondrial Injury by Enhanced Expression of Heme Oxygenase-1 in HT-22 Neuronal Cells (NMDA를 처리한 HT-22 신경세포에서 미토콘드리아 손상을 완화하는 레스베라트롤의 보호 효과와 헴 산화효소-1의 역할)

  • Kang, Jae Hoon;Woo, Jae Suk
    • Journal of Life Science
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    • v.32 no.1
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    • pp.11-22
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    • 2022
  • N-methyl-D-aspartate (NMDA) receptors have received considerable attention regarding their involvement in glutamate-induced neuronal excitotoxicity. Resveratrol has been shown to exhibit neuroprotective effects against this kind of overactivation, but the underlying cellular mechanisms are not yet clearly understood. In this study, HT-22 neuronal cells were treated with NMDA in Mg2+-free buffer and subsequently used as an experimental model of glutamate excitotoxicity to elucidate the mechanisms of resveratrol-induced neuroprotection. We found that NMDA treatment causes a drop in MTT reduction ability, disrupts inside-negative transmembrane potential of mitochondria, depletes cellular ATP levels, and stimulates intracellular ROS production. Double fluorescence imaging studies demonstrated an increased formation of mitochondrial permeability transition (MPT) pores accompanied by apoptotic cell death, while cobalt protoporphyrin and bilirubin showed protective effects against NMDA-induced mitochondrial injury. On the other hand, zinc protoporphyrin IX significantly attenuated the protective effects of resveratrol which was itself shown to enhance heme oxygenase-1 (HO-1) mRNA and protein expression levels. In cells transfected with HO-1 small interfering RNA, resveratrol failed to suppress the NMDA-induced effects on MTT reduction ability and MPT pore formation. The present study suggests that resveratrol may prevent mitochondrial injury in NMDA- treated HT-22 cells and that enhanced expression of HO-1 is involved in the underlying cellular mechanism.

A study on the effect of microgroove-fibronectin complex titanium plate on the expression of various cell behavior-related genes in human gingival fibroblasts (인간치은섬유아세포의 다양한 세포행동 관련 유전자발현에 마이크로그루브-파이브로넥틴 복합 티타늄표면이 미치는 영향에 대한 연구)

  • Hwang, Yu Jeong;Lee, Won Joong;Leesungbok, Richard;Lee, Suk Won
    • Journal of Dental Rehabilitation and Applied Science
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    • v.38 no.3
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    • pp.150-161
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    • 2022
  • Purpose: To determine the effects of the microgroove-fibronectin complex surface on the expression of various genes related to cellular activity in human gingival fibroblasts. Materials and Methods: Smooth titanium specimens (NE0), acid-treated titanium specimens (E0), microgroove and acid-treated titanium specimens (E60/10), fibronectin-fixed smooth titanium specimens (NE0FN), acid-treated and fibronectin-immobilized titanium specimens (E0FN), and microgroove and acid-treated titanium specimens immobilized with fibronectin (E60/10FN) were prepared. Real-time polymerase chain reaction experiments were conducted on 44 genes related to cell behavior of human gingival fibroblasts. Results: Adhesion and proliferation of human gingival fibroblast on microgroove-fibronectin complex titanium were activated through four types of signaling pathway. Integrin α5, Integrin β1, Integrin β3, Talin-2, which belong to the focal adhesion pathway, AKT1, AKT2, NF-κB, which belong to the PI3K-AKT signaling pathway, MEK2, ERK1, ERK2, which belong to the MAPK signaling pathway, and Cyclin D1, CDK4, CDK6 genes belonging to the cell cycle signaling pathway were upregulated on the microgroove-fibronectin complex titanium surface (E60/10FN). Conclusion: The microgroove-fibronectin complex titanium surface can up-regulate various genes involved in cell behavior.

Correlation Between Angiotensin-Converting Enzyme(ACE) Inhibitor Induced Dry Cough and ACE Gene Insertion/Deletion(I/D) Polymorphism (안지오텐신 전환효소 억제제에 의한 건성 기침의 발생과 안지오텐신 전환효소 유전자 다형성과의 관계)

  • Kim, Je-Hyeong;Jeong, Hye-Cheol;Kim, Kyung-Kyu;Lee, Sung-Yong;Kwon, Young-Hwan;Lee, So-Ra;Lee, Sang-Youb;Lee, Sin-Hyung;Cha, Dae-Ryong;Cho, Jae-Youn;Shim, Jae-Jeong;Cho, Won-Yong;Kang, Kyung-Ho;Kim, Hyoung-Kyu;Yoo, Se-Hwa;In, Kwang-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.46 no.2
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    • pp.241-250
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    • 1999
  • Background: Persistent nonproductive cough is a major adverse effect encountered with ACE inhibitor treatment and the most frequent reason for withdrawal of the drug. The mechanism of cough was postulated to be associated with accumulation of bronchial irritants which are substrates of ACE. It has been speculated that occurrence of this adverse effect is genetically predetermined ; in particular, variants of the genes encoding ACE. To investigate this relationship, we determined ACE gene Insertion/Deletion polymorphism in subjects with and without a history of ACE inhibitor-induced cough. Methods: Among the 339 patients with ACE inhibitor treatment, subjects who developed cough that resolved when not taking medication were designated to cough group and other subjects who did not complain cough were designated to non-cough group. Clinical characteristics of the patients were collected by review of medical records. ACE genotypes were determined by PCR amplification of DNA from peripheral blood and agarose gel electrophoresis. Results: 37 patients complained of dry cough(cough group) and 302 patients did not complained of cough(non-cough group). The incidence of ACE inhibitor induced dry cough was 10.9%. There was a preponderance of females in the cough group (M : F=24.3% : 75.7%) compared to the non-cough group (M : F=49.7% : 50.3%, p=0.004). There was no significant difference in mean age, underlying diseases, and kinds and frequencies of ACE inhibitors and their mean dosage between the both groups. ACE genotypic frequencies were I/I : I/D : D/D=16.2% : 18.9% : 64.9% in the cough group and 18.9% : 18.2% : 62.9% in the non-cough group which showed no significant difference between the both groups(p=0.926). Allelic frequencies were I : D = 25.7% : 74.3% and 28.0% : 72.0% in the cough and non-cough group respectively and the difference was not significant(p = 0.676). Conclusion: The incidence of ACE inhibitor-induced cough are 10.9%, and women are more susceptible to ACE inhibitor-induced cough. ACE inhibitor-induced dry cough is not associated with ACE gene Insertion/Deletion polymorphism.

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Studies on Inheritance and Ecological Variation of the Culm Length and Its Related Characters in Short-Statured Rice Varieties (수도단간품종의 간장 및 관련형질의 유전과 생태적 변이에 관한 연구)

  • Sung-Ho Bea
    • KOREAN JOURNAL OF CROP SCIENCE
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    • v.13
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    • pp.1-40
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    • 1973
  • These studies were aimed at clarification of genetic and ecological variation in culm length, panicle length and plant height of the $\textrm{F}_2$ plants in some selected crosses made between semi-dwarf rice varieties and tall Japonica ones. One Indica semi-dwarf, Taichung Native 1, one Indica $\times$ Japonica hybrid, IE51 and one Japonica semi-dwarf, Tankanbaekmang were used as short-gene donors while two of medium maturity varieties, Jinheung and Kwanok and one late veriety, Palkweng were used as the corresponding counterpart of respective dwarf varieties in a series of crosses. Five different crosses, Kwanok $\times$ Tankanbaekmang, Palkweng $\times$ Tankanbaekmang, Jinheung $\times$ T(N)1, Kwanok $\times$ T(N)1 and Kwanok $\times$ IE51, were made among the above six varieties. The $\textrm{F}_2$ plants of these crosses together with the concerned parental varieties were grown under several different conditions including three levels of each nitrogen and planting space, three planting seasons and three locations in 1968, to investigate variation in length of culm and panicle, and plant height. On the other hand, the F$_3$ progenies which were derived from the shortest 10 percent of the plants of three $\textrm{F}_2$ populations, Kwanok $\times$ T(N)1, Jinheung $\times$ T(N) 1 and Kwanok $\times$ IE51 grown in the previous year, were compared each other on the basis of selection efficiency in culm length. The experimental results could be summarized as follows; 1. Genetic behavior A. It was revealed that Tankanbaekmang, one of Japonica dwarf has a simple recessive gene responsible for short culm expression, showing a typical segregation ratio of three tall to one short culm plants in $\textrm{F}_2$ generation of the crosses either with Kwanok or Palkweng. B. In the both combinations, segregation pattern of the panicle length was exactly same as that of culm length. It seems that the same gene controls both culm length and panicle length. C. No difference between segregation of culm length and plant height in the above crosses was observed. D. T(N)1, one of Indica semi-dwarf did not show such a simple genetic behavior as detected from the crosses with Tankanbaekmang in segregation of culm length but formed a continuous and normal distribution curve. Therefore, some nonallelic genic actions might be involved in expression of culm length of the counterpart varieties of T(N)1. In particular, a transgressive segregation appeared toward the direction of longer culm length in case of Jinheung $\times$ T(N)1. The genetic behavior of panicle length and plant height generally coincided with that of culm length in all the cases. E. IE51 demonstrated exactly the same genetic behavior as that of T(N)1 when this variety was crossed with Kwanok. It was clearly clarified that the simple recessive gene controlling dwarfism from T(N)1 was well incorporated into this variety. 2. Ecological variation A. In general, there was a decreasing tendency in culm length and plant height of rice plant as seeding delayed while it was not so noticeable in panicle length. The decreasing magnitude varied from variety to variety and from cross to cross. Genetic behavior of the culm length and related characters of these materials was not disturbed by the variation of seeding season, nitrogen level, planting space and experimental location. E. The elongation mode of the upper three internodes was very similar to the segregation mode of culm length, panicle length and plant height in $\textrm{F}_2$ populations of . all the crosses investigated in this study. Accordingly, this result confirmed that the roles of the upper three internodes are very important in manifesting plant stature in rice. C. The effect of nitrogen on culm length and the related other two characters seemed to be meager. However, it was true to show an increasing tendency of those characters as nitrogen level got increased from 4 kg to 12kg per l0a, with different magnitude depending upon variety or cross. D. Also, the effect of planting space on culm length, panicle length and plant height was relatively small in all the cases. Those characters varied again depending upon variety or cross. However, a general increasing tendency was detected in manifestation of those traits under denser planting space condition. E. All the parental varieties produced shorter culm, panicle and plant height when they were grown at the lower latitude locations. It might be attributed to the fact that their reproductive growth accelerated with increased temperature prevailing at the lower latitude locations such as Iri and Mi1yang. On the countrary, $\textrm{F}_2$ population reacted differently to the different locations from the parental varieties. All the $\textrm{F}_2$ plants produced the longest culm, panicle and plant at Milyang. 3. Selection efficiency A. The heritability of culm length in Kwanok $\times$ T(N)1, Kwanok $\times$ IE51 and Jinheung$\times$T(N)1 was 92 percent, 74 percent and 55 percent, respectively. B. The actual genetic advance for culm length obtained from the progeny lines of the selected plants(10 precent) from the $\textrm{F}_2$ generation, was comparable to the expected advance calculated from the original $\textrm{F}_2$ populations. As compared with the $\textrm{F}_2$ population, the $\textrm{F}_3$ plants of Kwanok $\times$ T(N)l shortened on the average by 20.8cm, those of Kwanok $\times$ IE51 did 8.7cm and those of Jinheung$\times$T(N)1 20.0cm, respectively. C. Panicle length of the populations was differently affected from one cross to another by the selection based upon culm length in $\textrm{F}_2$ Kwanok $\times$ T(N)1 did not show any noticeable shortening of its culm length due to the selection pressure. On the other hand, both Kwanok $\times$ IE51 and Jinheung $\times$ T(N)1 showed a considerable shortening of their panicles in case of selection for culm length. Based upon the above results, it could be concluded that the ecological variation in culm length, panicle length and plant height was relatively small and fallen within the range of genetic variation. Considering from the fact that the simple recessive gene governing short height of Tankanbaekmang always accompanied with some undesirable characters such as short panicle and extremely small grain, the short gene of T(N)1 seemed to be more useful as dwarf gene source since it did not carry short gene together with such undesirable traits.

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