• Title/Summary/Keyword: substitution

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The Progression of SARS Coronavirus 2 (SARS-CoV2): Mutation in the Receptor Binding Domain of Spike Gene

  • Sinae Kim;Jong Ho Lee;Siyoung Lee;Saerok Shim;Tam T. Nguyen;Jihyeong Hwang;Heijun Kim;Yeo-Ok Choi;Jaewoo Hong;Suyoung Bae;Hyunjhung Jhun;Hokee Yum;Youngmin Lee;Edward D. Chan;Liping Yu;Tania Azam;Yong-Dae Kim;Su Cheong Yeom;Kwang Ha Yoo;Lin-Woo Kang;Kyeong-Cheol Shin;Soohyun Kim
    • IMMUNE NETWORK
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    • v.20 no.5
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    • pp.41.1-41.11
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    • 2020
  • Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) is a positive-sense single-stranded RNA (+ssRNA) that causes coronavirus disease 2019 (COVID-19). The viral genome encodes twelve genes for viral replication and infection. The third open reading frame is the spike (S) gene that encodes for the spike glycoprotein interacting with specific cell surface receptor - angiotensin converting enzyme 2 (ACE2) - on the host cell membrane. Most recent studies identified a single point mutation in S gene. A single point mutation in S gene leading to an amino acid substitution at codon 614 from an aspartic acid 614 into glycine (D614G) resulted in greater infectivity compared to the wild type SARS-CoV2. We were interested in investigating the mutation region of S gene of SARS-CoV2 from Korean COVID-19 patients. New mutation sites were found in the critical receptor binding domain (RBD) of S gene, which is adjacent to the aforementioned D614G mutation residue. This specific sequence data demonstrated the active progression of SARS-CoV2 by mutations in the RBD of S gene. The sequence information of new mutations is critical to the development of recombinant SARS-CoV2 spike antigens, which may be required to improve and advance the strategy against a wide range of possible SARS-CoV2 mutations.

cSNP Identification and Genotyping from C4B and BAT2 Assigned to the SLA Class III Region (돼지 SLA class III 영역 내 C4B 및 BAT2의 cSNP 동정 및 이를 이용한 유전자형 분석)

  • Kim, J.H.;Lim, H.T.;Seo, B.Y.;Lee, S.H.;Lee, J.B.;Yoo, C.K.;Jung, E.J.;Jeon, J.T.
    • Journal of Animal Science and Technology
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    • v.49 no.5
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    • pp.549-558
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    • 2007
  • C4B and BAT2, assigned to the SLA class III region, were recently reported on relation with human diseases. The primers for RT-PCR and RACE-PCR for CDS analysis of these genes of pig were designed by aligning the CDSs of humans and mice from GenBank. After we amplified and sequenced with these primers and cDNAs, the full-length CDSs of pig were determined. The CDS lengths of C4B and BAT2 were shown as 5226 bp and 6501 bp. In addition, the identities of nucleotide sequences with human and mouse were 76% to 87%, and the identities of amino acids were 72% to 90%. After we carried out the alignment with determined CDSs in this study and pig genomic sequences from GenBank, the primers for cSNP detection in genome were designed in intron regions that flanked one or more exons. Then, we amplified and directly sequenced with genomic DNAs of six pig breeds. Four cSNPs from C4B and three 3 cSNPs from BAT2 were identified. In addition, amino acid substitution occurred in six cSNP positions except for C4248T of C4B. By the Multiplex-ARMS method, we genotyped seven cSNPs with DNA samples used for direct sequencing. We verified that this result was the same as that analyzed using direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS on two randomly selected DNA samples. The genotype of each sample showed the same result from both methods. Therefore, seven cSNPs were identified from C4B and BAT2 and could be used as the basic data for haplotype analysis of SLA class III region. Moreover, the Multiplex-ARMS method should be powerful for genotyping of genes assigned to the whole SLA region for the xenograft study.

Association Between the Polymorphism in the Promoter Region of Porcine A-FABP Gene and Growth Traits in Duroc Pigs (돼지 Duroc 품종에서 A-FABP promoter의 다형성과 성장형질의 연관성)

  • Han, Sang-Hyeon;Jo, In-Cheol;Lee, Jong-Eon;Kim, Hyo-Seon;Lee, Jeong-Gyu;Jeon, Jin-Tae;O, Mun-Yu;Go, Mun-Seok
    • Journal of Animal Science and Technology
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    • v.48 no.1
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    • pp.21-28
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    • 2006
  • A polymorphism was found in the promoter region of porcine adipocyte fatty acid binding protein gene(A-FABP) gene which plays a key role in the binding and transportation of free fatty acid in adipocyte and deposition of intramuscular fat. Mutation was detected a substitution(T406C) using SSCP analysis and subsequently confirmed by sequencing the fragment in Duroc pigs. This T-406C mutation might change the binding activity for transcription factor nuclear factor 1(NF1). In this population, this mutation was genotyped using HinfⅠRFLP, and found three kinds of genotypes(TT, TC, and CC) showing their frequencies of 42.3, 44.3, and 13.4%, respectively. We statistically analyzed the association between the A-FABP genotypes and growth traits and found that the body weights of the pigs containing 406C/(TC or CC) were heavier for the body weight at the age of 20 weeks than those containing genotype TT(P<0.05), but not for those at the age of 0, 3, and 10 weeks. Pigs containing genotype CC had also a higher value for the average daily gain and lower values for the date for 90kg of body weight and food conversion ratio than those of 406T/- genotype. In addition, without the significant difference of back fat thickness, there was a significant association between the existence of allele CC and lean meat and eye muscle area(P<0.05). As a result of this study, we suggest that the allele T406C in the promoter region of A-FABP gene play an important role in deposition of intramuscular fat and weight in the later growth period. This polymorphism will be an useful molecular marker for breeding of Duroc pigs.

Chemical Equilibria of Lanthanide {Ln(III)=Pr, Sm, Gd, Dy}-Macrocyclic Complexes with Auxiliary Ligands in $CH_3OH$(Part III); Study of the Coordination of Nitrogen-or Oxygen-Containing Bases ($CH_3OH$ 용매에서 란탄족 원소 {Ln(III)=Pr, Sm, Gd, Dy}-거대고리 착물과 보조리간드 간의 화학평형(제3보); 두자리 리간드(주게원자:N혹은 O)를 중심으로 고찰)

  • Byun, Jong-Chul;Park, Yu-Chul;Han, Chung-Hun
    • Journal of the Korean Chemical Society
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    • v.44 no.2
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    • pp.95-101
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    • 2000
  • Macroacyclic transition metal complexes such as $Cu(H_2L[A]).H_2O$, $Cu(H_2L[B]).H_2O$, CuFe(L[A]($NO_3$).$4H_2O$, CuFe(L[B])($NO_3$).$4H_2O$, [$CuGd(H_2L[A])(NO_3)_2](NO_3).2CH_3OH$, [CuGd($H_2L$[B])($NO_3)_2$]($NO_3).2CH_3OH were prepared from the corresponding hexadentate compartmental ligands, $H_4L[A]$ and $H_4L[B]$, which were obtained by the condensation of 2-hydroxy-3-hydroxymethy1-5-methyIbenzaldehyde(HHNNB) and ethylenediamine or l,3-diaminopropane. Ln-macrocyclic([20]DOTA) complexes,[Ln([20]DOTA)($NO_3)(H_2O)$]($NO_3$)2.$xH_2O${Ln(III)=Pr, Sm, Gd, Dy, which had been synthesized from 2,6-diformyl-p-cresol(DFPC), was placed in methanol for 2 days, and [Ln([20] DOTA)($NO_3)(CH_3OH)]^{2+}$ was formed The equilibrium constants (k) for the substitution of coordinated $CH_3OH$ in the Ln-[20]DOTA complexes by various bidentate auxiliary ligands, $L_a$(=o-phenylenediamine,1,10-phenanthroline, ethylenediamine,oxalicacid, malonic acid, acethylacetone) were determined by spectroscopic method at $25^{\circ}C$ and 0.1M $NaClO_4$.The pKa of auxiliary ligands is in the order of o-phenylenediamine < 1,10-phenanthroline < ethylene-diamine, oxalic acid < malonic acid < acethylacetone. However, the equilibrium constant(K) has shown thetrend of ethyleneiamine < 1,10-phenanthroline < o-phenylenediamine, acethylacetone < malonic acid < oxalic acid.

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Investigating the Partial Substitution of Chicken Feather for Wood Fiber in the Production of Wood-based Fiberboard (목질 섬유판 제조에 있어 도계부산물인 닭털의 목섬유 부분적 대체화 탐색)

  • Yang, In;Park, Dae-Hak;Choi, Won-Sil;Oh, Sei Chang;Ahn, Dong-uk;Han, Gyu-Seong;Oh, Seung Won
    • Korean Chemical Engineering Research
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    • v.56 no.4
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    • pp.577-584
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    • 2018
  • This study was conducted to investigate the potential of chicken feather (CF), which is a by-product in poultry industry, as a partial substitute of wood fiber in the production of wood-based fiberboard. Keratin-type protein constituted the majority of CF, and its appearance did not differ from that of wood fiber. When the formaldehyde (HCHO) adsorptivities of CF compared by its pretreatment type, feather meal (FM), which was pretreated CF with high temperature and pressure and then grounded, showed the highest HCHO adsorptivity. In addition, there was no difference between the adsorbed HCHO amounts, which was measured by dinitrophenylhydrazine method, of scissors-chopped CF and CF beated with an electrical blender. Mechanical properties and HCHO emission of medium-density fiberboards (MDF), which were fabricated with wood fiber and 5 wt% CF, beated CF or FM based on the oven-dried weight of wood fiber, were not influenced by the pretreatment type of CF. However, when the values compared with those of MDF made with just wood fiber, thickness swelling and HCHO emission of the MDF were improved greatly with the addition of CF, beated CF or FM. Based on the results, it might be possible to produce MDF with improved dimensional stability and low HCHO emission if CF, beated CF or FM is added partially as a substitute of wood fiber in the manufacturing process of MDF produced with the conventional urea-formaldehyde resin of $E_1$ grade. However, the use of CF or FM in the production of MDF has a low economic feasibility at the current situation due to the securing difficulty and high cost of CF. In order to enhance the economic feasibility, it requires to use CF produced at small to medium-sized chicken meat plants. More importantly, it is considered that the technology developed from this research has a great potential to make provision for the prohibition of animal-based feed and to dispose environmentally avian influenza-infected poultry.

Interpretation on the Formative Design for Garden Pond of Hwaseol-dang in Muan (무안 화설당(花雪堂) 지당(池塘)의 조형디자인적 해석(解釋))

  • Rho, Jae-Hyun;Lee, Hyun-Woo
    • Journal of the Korean Institute of Traditional Landscape Architecture
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    • v.33 no.2
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    • pp.1-11
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    • 2015
  • This study sheds light on a pond design process which is a core facility of Hwaseol-dang in Muan, the Jeonnam. The plasticity of the pond was analyzed and interpreted for the design process using methods such as "literature search, interview, site visits, aerial pictures, aerial photographing, drawing figures of configuration plane via measurements, internet search, etc.", to trace the developing process of the design and the implications therein. The study results being centered on the developing process of the pond design are summarized herein below. The position of the Hwaseol-dang, being formed on a low hill having low competence as a place for a pavilion, draws more attention regarding its implications from the aspect of inner design. The pond Hwaseol-dang is in a rectangular shape of 1 : 1.2 ratio, in which the depth is a bit higher on the pond edge of the Hwaseol-dang thus being slanted, and Crape Myrtle, which is not known whether introduced during the formation of the pond, is cultivated on the island in the center widespread toward the southeast region. The planar design of the pond is interpreted as "rectangular pond" but it has a smooth half-moon shape where a part is excluded to remove edge. In particular, the three islands in rectangular pond, due to the narrow area, put one island and two half-moon-shaped islands in juxtaposition, and thus, although only being one island, resultantly exhibits the existence effect of proliferated three islands. This is allegedly due to the intentional formation aiming at the effect of hybrid while minimizing the overlap due to merging and adding from the aspect of constituting a design. Furthermore, the pond Hwaseol-dang is extended northwest along with Hwaseol-dang, and also the island in the center is thought to additionally have one or two, but the widespread phenomenon of the island in the center appears to consider the effect of "sit view on the floor of the pavilion of Hwaseol-dang". Considering that even a few examples of ponds having the three islands among the private house gardens in the nation are all curved ponds, the characteristics of the rectangular Hwaseol-dang pond establishing the garden effect of the three islands by modifying the one island in rectangular pond is highly notable. Considering that the three islands of "Yeongju, Bangjang, and Bongrae" is the original shape of the pond garden gestating Taoist ideology, as a symbolic design of a pond, it is regarded as the characteristics of the pond shape in Jeonnam area, and the so-called three treasures "Hwaseol-dang, Camellia, and oddly shaped stones, etc." are concentrated as the symbolism of Hwaseol-dang pond.

Comparison of PCR-RFLP and Real-Time PCR for Allelotyping of Single Nucleotide Polymorphisms of RRM1, a Lung Cancer Suppressor Gene (폐암 억제유전자 RRM1의 단일염기다형성 검사를 위한 PCR-RFLP법과 Real-Time PCR법의 유용성 비교)

  • Jeong, Ju-Yeon;Kim, Mi-Ran;Son, Jun-Gwang;Jung, Jong-Pil;Oh, In-Jae;Kim, Kyu-Sik;Kim, Young-Chul
    • Tuberculosis and Respiratory Diseases
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    • v.62 no.5
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    • pp.406-416
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    • 2007
  • Background: Single nucleotide polymorphisms (SNPs), which consist of a substitution of a single nucleotide pair, are the most abundant form of genetic variations occurring with a frequency of approximately 1 per 1000 base pairs. SNPs by themselves do not cause disease but can predispose humans to disease, modify the extent or severity of the disease or influence the drug response and treatment efficacy. Single nucleotide polymorphisms (SNPs), particularly those within the regulatory regions of the genes often influence the expression levels and can modify the disease. Studies examining the associations between SNP and the disease outcome have provided valuable insight into the disease etiology and potential therapeutic intervention. Traditionally, the genotyping of SNPs has been carried out using polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP), which is a low throughput technique not amenable for use in large-scale SNP studies. Recently, TaqMan real-time PCR chemistry was adapted for use in allelic discrimination assays. This study validated the accuracy and utility of real-time PCR technology for SNPs genotyping Methods: The SNPs in promoter sequence (-37 and -524) of lung cancer suppressor gene, RRM1 (ribonucleotide reductase M1 subunit) with the genomic DNA samples of 89 subjects were genotyped using both real-time PCR and PCR-RFLP. Results: The discordance rates were 2.2% (2 mismatches) in -37 and 16.3% (15 mismatches) in -524. Auto-direct sequencing of all the mismatched samples(17 cases) were in accord with the genotypes read by real-time PCR. In addition, 138 genomic DNAs were genotyped using real-time PCR in a duplicate manner (two separated assays). Ninety-eight percent of the samples showed concordance between the two assays. Conclusion: Real-time PCR allelic discrimination assays are amenable to high-throughput genotyping and overcome many of the problematic features associated with PCR-RFLP.

The Optimal Condition of Performing MTT Assay for the Determination of Radiation Sensitivity (방사선 감수성 측정법으로서 MTT 법 시행 시의 최적 조건에 대한 연구)

  • Hong, Se-Mie;Kim, Il-Han
    • Radiation Oncology Journal
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    • v.19 no.2
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    • pp.163-170
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    • 2001
  • Purpose : The measurement of radiation survival using a clonogenic assay, the established standard, can be difficult and time consuming. In this study, We have used the MTT assay, based on the reduction of a tetrazolium salt to a purple formazan precipitate by living cells, as a substitution for clonogenic assay and have examined the optimal condition for performing this assay in determination of radiation sensitivity. Materials and Methods : Four human cancer cell lines - PCI-1, SNU-1066, NCI-H630 and RKO cells have been used. For each cell line, a clonogenic assay and a MTT assay using Premix WST-1 solution, which is one of the tetrazolium salts and does not require washing or solubilization of the precipitate were carried out after irradiation of 0, 2, 4, 6, 8, 10 Gy. For clonogenic assay, cells in $25\;cm^2$ flasks were irradiated after overnight incubation and the resultant colonies containing more than 50 cells were scored after culturing the cells for $10\~14$ days. For MTT assay, the relationship between absorbance and cell number, optimal seeding cell number, and optimal timing of assay was determined. Then, MTT assay was performed when the irradiated cells had regained exponential growth or when the non-irradiated cells had undergone four or more doubling times. Results : There was minimal variation in the values gained from these two methods with the standard deviation generally less than $5\%$, and there were no statistically significant differences between two methods according to t-test in low radiation dose (below 6 Gy). The regression analyses showed high linear correlation with the $R^2$ value of $0.975\~0.992$ between data from the two different methods. The optimal cell numbers for MTT assay were found to be dependent on plating efficiency of used cell line. Less than 300 cells/well were appropriate for cells with high plating efficiency (more than $30\%$). For cells with low plating efficiency (less than $30\%$), 500 cells/well or more were appropriate for assay. The optimal time for MTT assay was after 6 doubling times for the results compatible with those of clonogenic assay, at least after 4 doubling times was required for valid results. In consideration of practical limits of assay (12 days, in this study) cells with doubling time more than 3 days were inappropriate for application. Conclusion : In conclusion, it is found that MTT assay can successfully replace clonogenic assay of tested cancer cell lines after irradiation only if MTT assay was undertaken with optimal assay conditions that included plating efficiency of each cell line and doubling time at least.

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Effect of Shift Interval for the Clinical Nurse on the Circadian Rhythm (임상 간호사의 교대근무 기간이 circadian rhythm 변화에 미치는 영향)

  • 황애란;정현숙;임영신;이혜원;김조자
    • Journal of Korean Academy of Nursing
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    • v.21 no.2
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    • pp.129-149
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    • 1991
  • Circadian rhythm is entrained in the 24-hour time interval by periodic factors in the environment, known as zeitgeber. But most rotating work schedules are outside the range of the entrainment of the pacemaker timing the human circadian sleep - wake cycle. It has been postulated that physiological and emotional disturbances occur in most human functions when the circadian rhythm is disturbed. So application of circadian principles to the design of shift schedules can aid in maintaining the temporal integrity of the circadian system and thereby minimize for the shift worker any detrimental consequences of circadian disruption. This study was a quasi-experimental study to test the effect of shift intervals for the clinical nurse on the circadian rhythm. Twenty nurses newly employed in general units of two hospitals were selected as an experimental group and twelve college nursing students as a control group. Both groups were selected according to an established criteria using a purposive sampling technique. Ten subjects were assigned to a weekly shift group and another ten to a biweekly shift group engaged in a semi -continuous shift schedule(sunday off) with a backward direction, that is, morning -evening - night shift. The control group worked a morning shift for 42 days. Oral temperature rhythm, waking tim, sleep - wake cycle, fatigue, and mental performance were measured during the experimental period. The data collection period was from April 30, 1990 to June 10, 1990. MANOVA, paired t-test, ANOVA, and Student Newman Keuls method were used for statistical analysis. The results are summarized as follows. 1. Phase delay in the acrophase of temperature rhythm was shown according to the backward rotating shift. A complete adaptation to work on the night shift was achieved between the sixth and ninth day of the night shift. 2. There was no difference in either waking time or sleep- wake cycle according to the duration of the working day for every shift group. Significant difference was found in the waking time and the sleep -wake cycle for subjects on the morning, evening, and night shift in both of the shift groups(weekly shift group : λ=0.121, p<0.01, λ=0.112, p<0.01, biweekly shift group : λ=0.116, p<0.01, λ=0.084, p<0.01). 3. There was no difference in fatigue between the first working day and the last working day for the control group and for the biweekly shift group. In the weekly shift group, physical fatigue was significantly different for the first day and the sixth day of the night shift(t=-2.28, p<0.05). Physical fatigue and total fatigue on the first day of the night shift showed a significant difference among the control group, the weekly shift group, and the biweekly shift group(F=5.79, p<0.01, F=4.56, p<0.05). There was a significant difference between the shift groups and the control group(p<0.05), Physical fatigue, neurosensory fatigue and total fatigue on the last day of the night shift showed a significant difference among the control group, the weekly shift group, and the biweekly shift group(F=12.65, p<0.01, F=7.77, p<0.01, F=9.68, p<0.01). There was a significant difference between the shift groups and the control group(p<0.05). 4. No difference in mental performance was seen between the first day and the last day of work in each case. An arithmatic test on the first day of the night shift revealed a significant difference among the control group, the weekly shift group, and the biweekly shift group(F=3.79, p<0.05). There was a significant difference between the shift groups and the control group(p<0.05) . The digital symbol substitution test and the arithmetic test on the last day of the night shift showed a significant difference among the control group, the weekly shift group, and the biweekly shift group(F=3.68, p<0.05, F=5.55, p<0.01), and both showed a significant difference between the shift groups and the control group(p<0.05). Accordingly, this study showed that during night duty, the waking time, sleep- wake cycle, and fatigue increased and mental performance decreased compared with morning and evening duty. It was also found that the weekly shift group had a higher fatigue score on the sixth day of night duty as compared to the -first day, but the waking time, sleep- wake cycle, and mental performance revealed no difference for the duration of the night duty or between shift groups, and complete adaptation of temperature rhythm was achieved between the sixth and ninth day of night duty. It is possible to conclude from these results that for intermediate circadian type in a healthy young woman, a biweekly shift system is more compatible with the circadian timing system than weekly shift system.

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Correlation Between Primary Tuberculous Pleurisy and NRAMP1 Genetic Polymorphism (결핵성 흉막염 환자에서 NRAMP1 유전자 다형성에 대한 연구)

  • Kim, Je-Hyeong;Kim, Byung-Gyu;Jung, Ki-Hwan;Park, Sang-Myun;Lee, Sang-Youb;Lee, Sin-Hyung;Sin, Cheol;Cho, Jae-Youn;Shim, Jae-Jeong;In, Kwang-Ho;Yoo, Se-Hwa;Kang, Kyung-Ho
    • Tuberculosis and Respiratory Diseases
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    • v.48 no.2
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    • pp.155-165
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    • 2000
  • Background: The phagolysosomal function of alveolar macrophage against M. tuberculosis infection is influenced by Nramp1, which is encoded by the NRAMP1 gene. There are several genetic polymorphisms in NRAMP1, and these polymorphisms affect the innate host resistance through the defect in production and function of Nramp1. To investigate this relationship, the NRAMP1 genetic polymorphism in patients with primary tuberculous pleurisy was determined. Methods: Fifty-six primary tuberculous pleurisy patient, who were diagnosed by pleural biopsy, were designated to the pleurisy group and 45 healthy adults were designated to the healthy control group. Three genetic polymorphisms of NRAMP1, such as a single point mutation in intron 4(469+14G/C, INT4), a nonconservative single-base substitution at codon 543 that changes aspartic acid to asparagine(D543N) and a TGTG deletion in the 3' untranslated region(1729+55delI4, 3'UTR), were determined. Polymerase chain reaction(PCR) and polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) were used. Results: The frequencies of mutant genotypes of INT4 and 3'UTR were significantly high in pleurisy group(p=0.001, p=0.023). But the frequencies of D543N were not significantly different between the two groups(p=0.079). The odds ratios, which are a comparison with wild genotype for determining mutant genotypes, were 8. 022(95% confidence interval=2.422-26.572) for INT4 and 5.733(95% confidence interval = 1.137~28.916) for 3'UTR ; these were statistically significant But the ratio for D543N was not significant In the combined analysis of the INT4 and 3'UTR polymorphisms, the odds ratios were 6.000(95% confidence interval = 1.461~24.640) for GC/++ genotype and 14.000(95% confidence interval=1.610~121.754) for GC/+del when compared with GG/++ homozygotes ; these were statistically significant. Conclusion: Among the NRAMP1 genetic polymorphisms, a single point mutation in intron 4(469+14G/C, INT4) and a TGTG deletion in the 3' untranslated region(1729+55del4, 3'UTR) were closely related to the primary tuberculous pleurisy.

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