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Evaluation of Candidate Housekeeping Genes for the Normalization of RT-qPCR Analysis using Developing Embryos and Prolarvae in Russian Sturgeon Acipenser gueldenstaedtii (러시아 철갑상어(Acipenser gueldenstaedtii) 발생 시료의 RT-qPCR 분석을 위한 내재 대조군 유전자의 선정)

  • Nam, Yoon Kwon;Lee, Sang Yoon;Kim, Eun Jeong
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.51 no.1
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    • pp.95-106
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    • 2018
  • To evaluate appropriate reference genes for the normalization of quantitative reverse transcription PCR (RT-qPCR) data with embryonic and larval samples from Russian sturgeon Acipenser gueldenstaedtii, the expression stability of eight candidate housekeeping genes, including beta-actin (ACTB), elongation factor-1A (EF1A), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), histone 2A (H2A), ribosomal protein L5 (RPL5), ribosomal protein L7 (RPL7), succinate dehydrogenase (SDHA), and ubiquitin-conjugating enzyme E2 (UBE2A), were tested using embryonic samples from 12 developmental stages and larval samples from 11 ontogenic stages. Based on the stability rankings from three statistic software packages, geNorm, NormFinder, and BestKeeper, the expression stability of the embryonic subset was ranked as UBE2A>H2A>SDHA>GAPDH>RPL5>EF1A>ACTB>RPL7. On the other hand, the ranking in the larval subset was determined as UBE2A>GAPDH>SDHA>RPL5>RPL7>H2A>EF1A>AC TB. When the two subsets were combined, the overall ranking was UBE2A>SDHA>H2A>RPL5>GAPDH>EF1A>ACTB>RPL7. Taken together, our data suggest that UBE2A and SDHA are recommended as suitable references for developmental and ontogenic samples of this sturgeon species, whereas traditional housekeepers such as ACTB and GAPDH may not be suitable candidates.

Isolation and Characterization of Pyrimidine Auxotrophs from the Hyperthermophilic Archaeon Sulfolobus acidocaldarius DSM 639 (Sulfolobus acidocaldarius 균주로부터 피리미딘 영양요구주의 분리 및 특성 연구)

  • Choi, Kyoung-Hwa;Cha, Jae-Ho
    • Journal of Life Science
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    • v.21 no.10
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    • pp.1370-1376
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    • 2011
  • To study the functional genomic analysis of a crenachaeon Sulfolobus acidocaldarius, we have constructed an auxotrophic mutant based on pyrEF, which encodes the pyrimidine biosynthetic enzymes orotate phosphoribosyltransferase and orotidine-5'-monophosphate decarboxylase. S. acidocaldarius was shown to be sensitive to 5-fluoroorotic acid (5-FOA), which can be selected for mutations in pyrEF genes within a pyrimidine biosynthesis cluster. Spontaneous 5-FOA-resistant mutants by ultraviolet, KH1U and KH2U, were found to contain two point mutations and a frame shift mutation in pyrE, respectively. Mutations at these sites from KH1U and KH2U decreased the activity of orotate phosphoribosyltransferase encoded by the pyrE gene and blocked the degradation of 5-FOA into toxic 5-FOMP and 5-FUMP that kill the cells. Therefore, KH1U and KH2U were uracil auxotrophs. Transformation of Sulfolobus-Escherichia coli shuttle vector pC bearing pyrEF genes from S. solfataricus P2 into S. acidocaldarius mutant KH2U restored 5-FOA sensitivity and overcame the uracil auxotrophy. This study establishes an efficient genetic strategy towards the systematic knockout of genes in S. acidocaldarius.

Differential expression of soybean SLTI100 gene encoding translation elongation factor 1A by abiotic stresses

  • Chung, Eun-Sook;Cho, Chang-Woo;So, Hyun-A;Yun, Bo-Hyun;Lee, Jai-Heon
    • Journal of Plant Biotechnology
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    • v.36 no.3
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    • pp.255-260
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    • 2009
  • The translation elongation factor 1A, eEF1A, catalyzes the binding of aminoacyl-tRNA to the A-site of the ribosome by a GTP-dependent mechanism. By subtractive suppression hybridization technique, we have isolated a soybean low-temperature inducible gene, SLTI100 encoding translation elongation factor 1A. Multiple sequence alignments and phylogenic analysis showed that SLTI100 and other eEF1As originated from diverse organisms are highly conserved. RNA expression of SLTI100 was specifically induced by low temperature, high salt, ABA, or drought stress. Based on the subcellular localization of the corresponding gene product fused to GFP, we were able to confirm that SLTI100-GFP was restricted to the nucleus and cytoplasm. We propose that soybean eEF1A may play an important role in translational regulation during abiotic stress responses in plants.

A Case of Eosinophilic Fasciitis Presenting as Stiffness of all Limbs (사지의 강직을 주소로 내원한 호산구성 근막염 환자)

  • Heo, Jae-Hyeok;Min, Ju-Hong;Kwon, Hyung-Min;Kim, Ji-Young;Cho, Joong-Yang;Lee, Kwang-Woo
    • Annals of Clinical Neurophysiology
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    • v.7 no.1
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    • pp.46-48
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    • 2005
  • Eosinophilic fasciitis (EF), also known as Shulman syndrome, is an inflammatory disorder of unknown etiology. It usually presents with pain, swelling, and tenderness of the proximal aspect of the limbs, chest, or neck, with subsequent induration of the skin and subcutaneous tissues, in association with peripheral eosinophilia. EF is differentiated from scleroderma by the pattern of skin involvement and non-involvement of muscle. We report a case of progressive EF presented with tightness and stiffness in all limbs.

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Effect of Application on the Growth of Head Lettuce under Fertigation Conditions in Highland (고랭지 결구상추의 관비재배시 돈분뇨 발효액비 시용효과)

  • 김원배;권영기;장석우;임상철
    • Korean Journal of Plant Resources
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    • v.14 no.1
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    • pp.84-88
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    • 2001
  • The growth of leaves in field-grown head lettuce(cv. Urake) was compared after conventional application of N-P-K fertilizer or after application of various dilutions of livestock waste solution. Plants treated with livestock waste solution diluted 25 times (Ef. 25) or 50 times (Ef. 50) had greater growth than that for any other treatment. The number of leaves was greater with Ef. 25 than that with other treatments, although the increase was not statistically significant. Total head yield increased by fertigation with livestock waste solution compared to yield resulting from conventional fertilization. Plant tissue weight was increased according to the increase in concentration of applied livestock waste solution. The soil chemical properties, pH, electrical conductivity, $NH_4^+-N$, NO$_3$-N, available P exchangeable Ca and Mg, all increased with the increased application of livestock waste solution. In conclusion, fertigation of the dilute livestock waste solution on the range of Ef. 25 to Ef. 50 could improve yield and quality in head lettuce.

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Investigation of right ventricle function in patients with tetralogy of Fallot after total correction using cardiac magnetic resonance imaging (심장 자기공명영상을 이용한 팔로사징 완전 교정술 후 우심실 기능에 대한 연구)

  • Jang, Woo Sung;Choi, Hee Joung;Lee, Jong-Min;Kim, Jae Bum;Kim, Jae Hyun;Jang, Jae Seok
    • Journal of Yeungnam Medical Science
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    • v.34 no.2
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    • pp.238-241
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    • 2017
  • Background: We investigated the difference in right ventricle (RV) volume and ejection fraction (EF) according to the pulmonary valve (PV) annular extension technique during Tetralogy of Fallot (TOF) total correction. Methods: We divided patients who underwent the procedure from 1993 to 2003 into two groups according to PV extension technique (group I: PV annular extension, group II: no PV annular extension) during TOF total correction. We then analyzed the three segmental (RV inlet, trabecular and outlet) and whole RV volume and EF by cardiac magnetic resonance imaging (MRI). Results: Fourteen patients were included in this study (group I: 10 patients, group II: four patients; male: nine patients, female: five patients). Cardiac MRI was conducted after a 16.1 years TOF total correction follow-up period. There was no statistical difference in RV segmental volume index or EF between groups (all p>0.05). Moreover, the total RV volume index and EF did not differ significantly between groups (all p>0.05). Conclusion: The RV volume and EF of the PV annular extension group did not differ from that of the PV annular extension group. Thus, PV annular preservation technique did not show the surgical advantage compared to PV annular extension technique in this study.

Evaluating Usefulness of Deep Learning Based Left Ventricle Segmentation in Cardiac Gated Blood Pool Scan (게이트심장혈액풀검사에서 딥러닝 기반 좌심실 영역 분할방법의 유용성 평가)

  • Oh, Joo-Young;Jeong, Eui-Hwan;Lee, Joo-Young;Park, Hoon-Hee
    • Journal of radiological science and technology
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    • v.45 no.2
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    • pp.151-158
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    • 2022
  • The Cardiac Gated Blood Pool (GBP) scintigram, a nuclear medicine imaging, calculates the left ventricular Ejection Fraction (EF) by segmenting the left ventricle from the heart. However, in order to accurately segment the substructure of the heart, specialized knowledge of cardiac anatomy is required, and depending on the expert's processing, there may be a problem in which the left ventricular EF is calculated differently. In this study, using the DeepLabV3 architecture, GBP images were trained on 93 training data with a ResNet-50 backbone. Afterwards, the trained model was applied to 23 separate test sets of GBP to evaluate the reproducibility of the region of interest and left ventricular EF. Pixel accuracy, dice coefficient, and IoU for the region of interest were 99.32±0.20, 94.65±1.45, 89.89±2.62(%) at the diastolic phase, and 99.26±0.34, 90.16±4.19, and 82.33±6.69(%) at the systolic phase, respectively. Left ventricular EF was calculated to be an average of 60.37±7.32% in the ROI set by humans and 58.68±7.22% in the ROI set by the deep learning segmentation model. (p<0.05) The automated segmentation method using deep learning presented in this study similarly predicts the average human-set ROI and left ventricular EF when a random GBP image is an input. If the automatic segmentation method is developed and applied to the functional examination method that needs to set ROI in the field of cardiac scintigram in nuclear medicine in the future, it is expected to greatly contribute to improving the efficiency and accuracy of processing and analysis by nuclear medicine specialists.

Comparison of RAPD, AFLP, and EF -1 α Sequences for the Phylogenetic Analysis of Fusarium oxysporum and Its formae speciales in Korea

  • Park, Jae-Min;Kim, Gi-Young;Lee, Song-Jin;Kim, Mun-Ok;Huh, Man-Kyu;Lee, Tae-Ho;Lee, Jae-Dong
    • Mycobiology
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    • v.34 no.2
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    • pp.45-55
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    • 2006
  • Although Fursarium oxysporum causes diseases in economically important plant hosts, identification of F. oxysporum formae speciales has been difficult due to confusing phenotypic classification systems. To resolve these complexity, we evaluated genetic relationship of nine formae speciales of F. oxysporum with random amplified polymorphic DNA (RAPD), amplified fragment length polymorphism (AFLP), and translation elongation factor-l alpha ($EF-1{\alpha}$) gene. In addition, the correlation between mycotoxin content of fusaric acid and isolates based on molecular marker data was evaluated using the modified Mantel's test. According to these result, these fusaric acid-producing strains could not identify clearly, and independent of geographic locations and host specificities. However, in the identification of F. oxysporum formae speciales, especially, AFLP analysis showed a higher discriminatory power than that of a the RAPD and $EF-1{\alpha}$ analyses, all three techniques were able to detect genetic variability among F. oxysporum formae speciales in this study.

A Study of Titanium Phase Transition through In-situ EF-TEM Heating Experiments (EF-TEM 직접가열 실험을 통한 titanium의 고온 상전이 연구)

  • Kim, Jin-Gyu;Lee, Young-Bu;Kim, Youn-Joong
    • Applied Microscopy
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    • v.33 no.1
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    • pp.49-58
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    • 2003
  • The ${\alpha}-{\beta}$ phase transition of titanium was investigated through in-situ EF-TEM heating experiments. Three different areas of a titanium foil were observed to minimize statistical errors. Systematic recording of diffraction patterns and images was carried out from $RT{\rightarrow}600^{\circ}C{\rightarrow}900^{\circ}C{\rightarrow}RT$ on each area. The following results were obtained: (1) Transition of titanium takes place very rapidly at $900^{\circ}C$. Two phases of titanium, ${\alpha}\;and\;{\beta}$, coexist at this temperature. (2) The transited ${\beta}$-phase appears in the form of twinned plates which are arranged in rotation relationship one another. (3) Analyses of electron diffraction patterns and EDS data indicate that the thermal oxidation layer is gradually formed on the surface of titanium above $900^{\circ}C$, which hinders the reversible ${\beta}{\rightarrow}{\alpha}$ phase transition upon cooling.

Antithrombosis, Antidiabetes, and Antioxidant Activities of Houttuynia cordata (어성초의 항혈전, 항당뇨 및 항산화 활성)

  • Yun-Jin, Lee;Deok-Gyeong, Kang;Jong Sik, Kim;Man-Hyo, Lee;Ho-Yong, Sohn
    • Journal of Life Science
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    • v.33 no.1
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    • pp.43-49
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    • 2023
  • Houttuynia cordata belongs to the Saururacease family and its leaves, stems, and roots have been used as oriental medicines to treat pneumonia, acute or chronic bronchitis, enteritis, and abscesses and to remove extravasated blood. Recently, the antioxidant, anti-inflammation, antibacterial, and anti-proliferation activities and protection abilities of H. cordata against liver and neuron cell damage have been reported. In this study, ethanol extract and its solvent fractions (fractions of hexane, ethyl acetate, butanol, and water residue) were prepared, and their antithrombosis, antidiabetes, antioxidant, and hemolysis activities were evaluated. The ethyl-acetate fraction of H. cordata (EF-HC) showed the highest polyphenol and flavonoids contents among the fractions and exhibited strong antithrombosis and antioxidant activities. The EF-HC at 5 mg/ml showed 2.09-folds of thrombin time, 2.19-folds of prothrombin time, and 1.69-folds of activated partial thromboplastin time compared to the their solvent control and 30.9, 19.9, and 49.6 ㎍/ml of RC50 against DPPH, ABTS, and nitrite radicals, respectively. Furthermore, the EF-HC did not show any hemolytic activity up to 1 mg/ml, whereas the hexane fraction of H. cordata showed 55% hemolysis at 1 mg/ml. This is the first report of the antithrombosis activity of H. cordata. Our results suggest that quercitirin, hyperoside, orientin, and isoquercitrin in EF-HC are related to its antithrombosis and antioxidant activities and that the EF-HC could be developed as a promising antithrombosis agent.