• Title/Summary/Keyword: M2M Translation

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Effects of the cis-Acting Element in the 3' End of Porcine $\beta$-Casein Gene on the Expression in Mammary Epithelial Cells (돼지 $\beta$-Casein 유전자의 3' 말단 부위의 cis-Acting Element가 유선 상피 세포내의 발현에 미치는 영향)

  • Lee, Hwi-Cheul;Kim, Byoung-Ju;Byun, Sung-June;Lee, Seung-Hoon;Kim, Min-Ji;Chung, Hee Kyoung;Lee, Hyun-Gi;Jo, Su-Jin;Chang, Won-Kyong;Park, Jin-Ki;Lee, Poong-Yeon
    • Reproductive and Developmental Biology
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    • v.32 no.3
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    • pp.153-158
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    • 2008
  • Tissue-specific and temporal regulation of milk protein gene expression is advantageous when creating transgenic animal that produces foreign protein into milk. Gene expression, i.e. protein production, is regulated not only by promoter strength but also mRNA stability. Especially, poly A tail length by polyadenylation affects in vivo and in vitro mRNA stability and translation efficiency of the target gene. In the present study, nucleotide sequence of 3'-UTR was analyzed to evaluate the effects of mRNA stability on the target gene expression. Based on the poly A signal of 3' -untranslated region (UTR), nucleotide sequences of putative cytoplasmic polyadenylation elements (CPEs) and downstream elements (DSEs: U-rich, G-rich, GU-rich) were analyzed and used to construct 15 luciferase reporter vectors. Each vector was transfected to HC11 and porcine mammary gland cell (PMGC) and measured for dual luciferase expression levels after 48 hours of incubation. Luciferase expression was significantly higher in construct #6 (with CPE 2, 3 and DSE 1 of exon 9) and #11 (with CPE 2, 3 and DSE 1, 2 and 3 of exon 9) than construct #1 in the PMGC. These results suggest that expression of target genes in PMGC may be effectively expressed by using the construct #6 and #11 on production of transgenic pig.

Cloning and characterization of Giardia intestinalis cyclophilin

  • Yu, Hak-Sun;Kong, Hyun-Hee;Chung, Dong-Il
    • Parasites, Hosts and Diseases
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    • v.40 no.3
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    • pp.131-138
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    • 2002
  • The cyclophilins (Cyps) are family members of proteins that exhibit peptidylprolyl cis-trans isomerase (PPIase, EC 5.2.1.8) activity and bind the immunosuppressive agent cyclosprin A (CsA) in varying degrees. During the process of random sequencing of a cDNA library made from Giardia intestinalis WB strain, the cyclophilin gene (gicypl) was isolated. An open reading frame of gicyp1 gene was 576 nucleotides, which corresponded to a translation product of 176 amino acids (Gicypl). The identity with other Cyps was about 58-71%. The 13 residues that constituted the CsA binding site of human cyclophilin were also detected in the amino acid sequence of Gicypl, including tryptophan residue essential for the drug binding. The single copy of the gicypl gene was detected in the G. intestinalis chromosome by southern hybridization analysis. Recombinant Gicyp 1 protein clearly accelerated the rate of cis ${\rightarrow}$ trans isomerization of the peptide substrate and the catalysis was completely inhibited by the addition of $0.5{\;}{\mu}M$ CsA.

Purification and Characterization of an Antiviral Ribosome-inactivating Protein from Chenopodium album L.

  • Cho, Kang-Jin;Lee, Si-Myung;Kim, Yeong-Tae;Hwang, Young-Soo
    • Journal of Applied Biological Chemistry
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    • v.43 no.3
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    • pp.125-130
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    • 2000
  • An antiviral protein (CAP30) with ribosome-inactivating activity was purified from the leaves of Chenopodium album L. through ammonium sulfate precipitation and column chromatography using S-Sepharose, Blue-Sepharose, FPLC Suprose12 HR, and FPLC Mono-S. The molecular wight of CAP30 was estimated to be 30kD. CAP30 was thermostable, maintaing its activity even after incubation at $70^{\circ}C$ for 30 min, and was stable in the pH range of 6 to 9. In a cell-free in vitro translation system using rabbit reticulocyte lysate, protein synthesis was inhibited by the addition of CAP30 with an $IC_{50}$ of 2.26pM. The comparison of N-terminal amino acid sequences of this protein with known ribosome-inactivating proteins (RIPs) revealed that it had some sequence homology with PAP-S and PAP-R from pokeweed (Phytolacca americana)and dodecandrin from P. dodecandra, but had no sequence homology with RIPs from other plants belonging to different orders. The mosaic symptoms on tobacco leaves caused by cucumber mosaic virus infection was completely inhibited by 100 ng/ml of the pure CAP30 protein.

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Comparison of the Modified Brostrom Repair Technique with and without Augmentation Using Suture Tape for Chronic Ankle Instability (만성 족관절 불안정성을 가진 환자군에서 변형 브로스트롬 술식과 봉합 테이프를 추가한 술식 간의 결과 비교)

  • Gwak, Heui-Chul;Jung, Soo-Hwan;Kim, Jung-Han;Park, Dae-Hyun;Choo, Hye-Jung;Kim, Dae-Yoo
    • Journal of Korean Foot and Ankle Society
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    • v.26 no.1
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    • pp.40-47
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    • 2022
  • Purpose: The modified Broström repair (BR) technique has yielded good outcomes in patients with chronic ankle instability. This study compared clinical and radiological outcomes between two groups of patients who underwent modified BR or lateral ligament augmentation using suture tapes (ST). Materials and Methods: Seventy-seven patients (ST group [n=47], BR group [n=30]; body mass index <26.61 kg/m2; mean age, 30.7±11.0 years [range, 17~39 years]; mean follow-up, 34.0±12.0 months [range, 24~59 months]) were retrospectively reviewed between January 2014 and July 2017. The Foot and Ankle Outcome Score (FAOS), American Orthopedic Foot and Ankle Score (AOFAS), Foot and Ankle Ability Measure (FAAM), visual analogue scale (VAS) score, and Sefton grading system were used for clinical assessment. The talar tilt angle and anterior talar translation were measured using the Telos stress device (Telos GmbH, Marburg, Germany) at 150 N for radiological evaluation. Results: FAOS, AOFAS, FAAM, and VAS scores improved in both groups at final follow-up (ST, 91.1±5.2, 93±2, 88.1±4.5, 1.5±0.7 vs. BR, 91.3±5.4, 93±3, 83.3±4.8, 1.2±0.7, respectively; p=0.854, 0.971, <0.001, 0.04, respectively). According to the FAOS, mean sports activity scores for the ST and BR groups at the final follow-up were 90.3±3.2 and 76.6±4.2, respectively, reflecting superior outcomes in the ST group (p<0.001). Sefton grading revealed satisfactory functional outcomes (ST, 91.5% vs. BR, 90.0%). There was significant improvement in the talar tilt angle and anterior talar translation in both the ST and BR groups (7.6°±1.2°, 10.5±1.8 mm vs. 4.9°±1.1°, 7.9±1.5 mm, respectively; p<0.001). Conclusion: The ST group demonstrated comparable clinical but better improvement in mechanical stability and FAOS sports scores than the BR group.

Evaluation of the Usefulness of Exactrac in Image-guided Radiation Therapy for Head and Neck Cancer (두경부암의 영상유도방사선치료에서 ExacTrac의 유용성 평가)

  • Baek, Min Gyu;Kim, Min Woo;Ha, Se Min;Chae, Jong Pyo;Jo, Guang Sub;Lee, Sang Bong
    • The Journal of Korean Society for Radiation Therapy
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    • v.32
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    • pp.7-15
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    • 2020
  • Purpose: In modern radiotherapy technology, several methods of image guided radiation therapy (IGRT) are used to deliver accurate doses to tumor target locations and normal organs, including CBCT (Cone Beam Computed Tomography) and other devices, ExacTrac System, other than CBCT equipped with linear accelerators. In previous studies comparing the two systems, positional errors were analysed rearwards using Offline-view or evaluated only with a Yaw rotation with the X, Y, and Z axes. In this study, when using CBCT and ExacTrac to perform 6 Degree of the Freedom(DoF) Online IGRT in a treatment center with two equipment, the difference between the set-up calibration values seen in each system, the time taken for patient set-up, and the radiation usefulness of the imaging device is evaluated. Materials and Methods: In order to evaluate the difference between mobile calibrations and exposure radiation dose, the glass dosimetry and Rando Phantom were used for 11 cancer patients with head circumference from March to October 2017 in order to assess the difference between mobile calibrations and the time taken from Set-up to shortly before IGRT. CBCT and ExacTrac System were used for IGRT of all patients. An average of 10 CBCT and ExacTrac images were obtained per patient during the total treatment period, and the difference in 6D Online Automation values between the two systems was calculated within the ROI setting. In this case, the area of interest designation in the image obtained from CBCT was fixed to the same anatomical structure as the image obtained through ExacTrac. The difference in positional values for the six axes (SI, AP, LR; Rotation group: Pitch, Roll, Rtn) between the two systems, the total time taken from patient set-up to just before IGRT, and exposure dose were measured and compared respectively with the RandoPhantom. Results: the set-up error in the phantom and patient was less than 1mm in the translation group and less than 1.5° in the rotation group, and the RMS values of all axes except the Rtn value were less than 1mm and 1°. The time taken to correct the set-up error in each system was an average of 256±47.6sec for IGRT using CBCT and 84±3.5sec for ExacTrac, respectively. Radiation exposure dose by IGRT per treatment was measured at 37 times higher than ExacTrac in CBCT and ExacTrac at 2.468mGy and 0.066mGy at Oral Mucosa among the 7 measurement locations in the head and neck area. Conclusion: Through 6D online automatic positioning between the CBCT and ExacTrac systems, the set-up error was found to be less than 1mm, 1.02°, including the patient's movement (random error), as well as the systematic error of the two systems. This error range is considered to be reasonable when considering that the PTV Margin is 3mm during the head and neck IMRT treatment in the present study. However, considering the changes in target and risk organs due to changes in patient weight during the treatment period, it is considered to be appropriately used in combination with CBCT.

Hydrogen sulfide ameliorates abdominal aorta coarctation-induced myocardial fibrosis by inhibiting pyroptosis through regulating eukaryotic translation initiation factor 2α phosphorylation and activating PI3K/AKT1 pathway

  • Yaling Li;Zhixiong Wu;Jiangping Hu;Gongli Liu;Hongming Hu;Fan Ouyang;Jun Yang
    • The Korean Journal of Physiology and Pharmacology
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    • v.27 no.4
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    • pp.345-356
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    • 2023
  • This study aimed to assess the effects of exogenous hydrogen sulfide (H2S) on abdominal aorta coarctation (AAC) induced myocardial fibrosis (MF) and autophagy in rats. Forty-four Sprague-Dawley rats were randomly divided into control group, AAC group, AAC + H2S group, and H2S control group. After a model of rats with AAC was built surgically, AAC + H2S group and H2S group were injected intraperitoneally with H2S (100 µmol/kg) daily. The rats in the control group and the AAC group were injected with the same amount of PBS. We observed that H2S can improve left ventricular function and the deposition of myocardial collagen fibers, inhibit pyroptosis, down-regulate the expression of P-eif2α in myocardial tissue, and inhibit cell autophagy by activating the phosphatidylinositol 3-kinase (PI3K)/AKT1 signaling pathway (p < 0.05). In addition, angiotensin II (1 µM) H9c2 cardiomyocytes were injured in vitro experiments, and it was also observed that pyroptosis was inhibited after H2S (400 µmol/kg) intervention, the expression of P-eif2α in cardiomyocytes was significantly down-regulated, and the PI3K/AKT1 signaling pathway was activated at the same time. Therefore, increasing the expression of P-eif2α reverses the activation of the PI3K/AKT1 signaling pathway by H2S. In conclusion, these findings suggest that exogenous H2S can ameliorate MF in rats with AAC by inhibiting pyroptosis, and the mechanism may be associated with inhibiting the phosphorylation of eif2α and activating the PI3K/AKT1 signaling pathway to inhibit excessive cell autophagy.

CD Gene Microarray Profiles of Bambusae Caulis in Liquamen in Human Mast Cell

  • Jeon Hoon;Kang Nan Joo;Kim Gyo Seok
    • Journal of Physiology & Pathology in Korean Medicine
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    • v.17 no.1
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    • pp.241-246
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    • 2003
  • Bambusae Caulis in Liquamen(BCL) has been used to relieve the cough and asthma, and remove the phlegm in traditional Oriental medicine. In recent years, it was studied for its antiinflammatory, antiallergenic, immune-modulating, and anticarcinogenic capabilities. This experiment was performed to evaluate the microarray profiles of CD genes in human mast cells before and after BCL treatment. The results are as follows: The expression of 51 of the genes studied was up-regulated in the Bel-treated group; they include the genes coding L apoferritin, beta-2-microglobulin, ferritin light polypeptide, CD63, monocyte chemotactic and activating fact, heme oxygenase 1, CD140a, integrin alpha M, colony stimulating factor 2 receptor, eukaryotic translation elongation factor, CD37, interleukin 18, NADH dehydrogenase 1 beta, CD48, 5-lipoxygenase activating protein, interleukin 4, ribosomal protein L5, GABA(A) receptor-associated protein, beta-tubulin, integrin beta 1, CD162, CD32, lymphotoxin beta, alpha-tublin, integrin alpha L, CD2, CD151, CD331, 90 kDa heat shock protein, CD59, CD3Z, microsomal glutathione S-transferase 2, CD33, CD162R, cyclophilinA, CD84, interleukin 9 receptor, interleukin 11, CD117, CD39-Like 2, and so forth. The expression of 7 of the genes studied was down-regulated in the BCL-treated group; they include the genes coding con, CD238, SCF, CD160, CD231, CD24, and CD130. Consequently, the treatment of BCL on the human mast cells increased the expression of 51 genes and decreased the expression of 7 genes. These data would provide a fundamental basis to the traditional applications of Bambusae Caulis in Liquamen.

Fusarium Wilt Caused by Fusarium oxysporum on Passionfruit in Korea (Fusarium oxysporum에 의한 패션프루트 시들음병)

  • Joa, Jae-Ho;Choi, In-Young;Choi, Min-Kyung;Heo, Byong-Soo;Jang, Jong-Ok;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.24 no.1
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    • pp.75-80
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    • 2018
  • From 2014 to 2016, Fusarium wilt disease was found on fassionfruit in Iksan and Jeju, Korea. Symptoms included wilting of foliage, drying and withering of leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelia with 35 mm after one week. Macroconidia were falcate to almost straight, thin-walled and usually 2-3 septate. Microconidia were usually formed on monophialides of the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $3-12{\times}2.5-6{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of two molecular markers, internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$, the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of F. oxysporum on fassionfruit in Korea.

Superoxide Dismutase Gene Expression in the Endotoxin-Treated Rat Lung (내독소에 의한 백서 폐장의 Superoxide Dismutase 유전자 발현에 관한 연구)

  • Yoo, Chul-Gyu;Suh, Gee-Young;Kim, Young-Whan;Han, Sung-Koo;Shim, Young-Soo;Kim, Keun-Youl;Han, Yong-Chol
    • Tuberculosis and Respiratory Diseases
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    • v.41 no.3
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    • pp.215-221
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    • 1994
  • Background: It is well known that oxygen free radicals(OFR) play a vital role in the various type of acute lung injury. Among various antioxidant defense mechanisms, the superoxide dismutases(SOD) are thought to be the first line of antioxidant defense by catalyzing the dismutation of two superoxide radicals to yield hydrogen peroxide and oxygen. Eukaryotic cells contain two types of intracellular SOD : cytosolic, dimeric copper/zinc- containing enzyme(CuZnSOD) and mitochondrial, tetrameric manganese-containing enzyme(MnSOD). The purpose of this study is to evaluate the time-dependent gene expression of MnSOD and CuZnSOD in the endotoxin-treated rats, and to compare with the manifestations of LPS-induced acute lung injury in rats. Methods: Total RNA from rat lung was isolated using single step phenol extraction 0, 1, 2, 4, 6, 12, 18, 24 hours after E. coli endotoxin injection(n=3, respectively). RNA was separated by formaldehyde-containing 1.2% agarose gels elctrophoresis, transblotted, baked, prehybridized, and hybridized with $^{32}P$-labeled cDNA probes for rat MnSOD and CuZnSOD, which were kindly donated by Dr. Ho(Duke University, Durham, NC, USA). The probes were labeled by nick translation. Blots were washed and autoradiography were quantitated using laser densitometry. Equivalent amounts of total RNA/gel were assessed by monitoring 28S and 18S rRNA. Results: Endotoxin caused a rise in steady-state MnSOD mRNA levels by 4h with peak mRNA accumulation by 6h. Continued MnSOD mRNA expression was observed at 12h. CuZnSOD mRNA expression was observed from 1h to 24h with peak levels by 18h. Conclusion: These results suggest that SOD palys an important defensive role in the endotoxin-induced acute lung injury in rats.

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First Report of Fusarium Wilt Caused by Fusarium oxysporum on Kohlrabi in Korea (Fusarium oxysporum에 의한 콜라비 시들음병)

  • Choi, In-Young;Kim, Ju;Ju, Ho-Jong;Park, Ji-Hyun;Shin, Hyeon-Dong
    • Research in Plant Disease
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    • v.21 no.1
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    • pp.27-31
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    • 2015
  • In April 2014, Fusarium wilt was found on kohlrabi seedlings in Iksan, Korea. Symptoms included wilting of foliage, drying and withering of older leaves, and stunting of the plants. The infected plants eventually died during growth. Colonies on potato dextrose agar were pinkish white, and felted with cottony and aerial mycelium. Macroconidia were falcate to almost straight, thin walled and usually 3-septate. Microconidia were usually formed abundantly in false-heads on short monophialides on the hyphae and were hyaline, smooth, oval to ellipsoidal, aseptate or medianly 1-septate, very occasionally 2-septate, slightly constricted at the septa, $4-11{\times}2.5-5{\mu}m$. On the basis of the morphological characteristics and phylogenetic analyses of molecular markers (internal transcribed spacer rDNA and translation elongation factor $1{\alpha}$), the fungus was identified as Fusarium oxysporum. Pathogenicity of a representative isolate was proved by artificial inoculation, fulfilling Koch's postulates. To our knowledge, this is the first report on the occurrence of Fusarium oxysporum on kohlrabi in Korea.