• Title/Summary/Keyword: C1-2 fusion

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Expression of Human Immunodeficiency Virus Type 1 Gag Protein in Escherichia coli

  • Park, Weon-Sang
    • Journal of Life Science
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    • v.9 no.5
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    • pp.556-563
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    • 1999
  • Presence of antibody to the capsid protein p24 is the main diagnostic criterion, since this reflects reliable antibody response to HIV infection. However, it takes about 6-8 weeks for antibody production after infection and people who are infected but antibodies are not produced yet are classified as seronegative. Therefore, there is a strong need for an improved diagnostic method for better health security. As a first step for developing such an improved diagnostic system, gag protein of human immunodificiency virus type 1 was expressed in E. coli DH5$\alpha$. The gag fragment of HIV-1 (including a portion of p17 and whole p24) was amplified by polymerase chain reaction (PCR) and BamH I/EcoR I sites were created during PCR. The amplified DNA fragment was cleaved with BamH I/EcoR I and was subcloned into the GEX-2T vector which had been digested with BamHI/EcoRI, resulting gene fusion with gst gene of pGEX-2T. The recombinant DNA was transferred into E. coli DH5$\alpha$. The transformed bacteria were grown at 37$^{\circ}C$ for 3h and protein expression was induced with 0.1mM IPTG at $25^{\circ}C$ for 3h. Recombinant gag protein or GST-gag fusion protein was purified with glutathione-sepharose 4B bead and migrated as a single band when analyzed by 10% polyacrylamide gel. These proteins were confirmed by immunoblotting with anti-GST goat sera or Korean AIDS patients sera. The results of this study establish the expression and single step pulification of HIV-1 gag protein which can specifically bind with Korean AIDS patients sera.

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A Study for Phoria and Positive Fusion Reverse on Vision Training of Convergence Insufficiency (폭주부족 피검사자의 시기능 훈련에 의한 양성융합여력 및 사위 변화에 관한 연구)

  • Lee, Chang-Sun;Kim, In-Su;Son, Jeong-Sik;Sung, Duk-Yong;Kim, Jaedo;Kim, Ki-Hong
    • Journal of Korean Ophthalmic Optics Society
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    • v.13 no.1
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    • pp.83-87
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    • 2008
  • Purpose: This study was investigated for the effects of vision training on the phoria and positive fusion reverse (PFR) of patients with convergence insufficiency (C.I). Methods: Subjects were included 11 patients with CI of T Vision Center (5 patients with distance orthophoria, 6 patients with distance exophoria). The vision training of this study had been processed daily during an average of 2 months for each patients. Results: The result showed that after vision training, the phoria and PFR of CI patient were improved significantly than that before vision training (p<0.05) and when the groups were divided to the distance orthophoria convergence insufficiency (DOCI) group and distance exophoria convergence insufficiency (DE-CI) group, these improvements were proved similarly in the both groups. Conclusions: This study proved that vision training for patients with C.I was effective for the improvement of the phoria and PFR of these patients.

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Strength and fracture toughness of reduced - activation ferritic steel (JLF-1) for fusion reactor application (핵융합로용 저방사화 철강재료(JLF-1)의 강도와 파괴인성)

  • Yun, Han-Gi;Kim, Dong-Hyeon;Lee, Sang-Pil;Park, Lee-Hyeon;Gong, Yu-Sik;Katoh, Y.;Kohyama, A.
    • Proceedings of the KSME Conference
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    • 2003.04a
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    • pp.13-18
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    • 2003
  • Reduced activation ferritic steel, JLF-1 steel (Fe-9Cr-2W-V-Ta), is one of the promising candidate materials for fusion reactor applications. Fracture toughness ($J_IC$) and tensile tests were carried out at room temperature and elevated temperature ($400^{\circ}C$). Two types of CT specimen were prepared to examine the effect of rolling direction on the fracture toughness of JLF-1 steel. Four types of tensile specimen were also prepared to investigate the property by the rolling direction and welding. The Micro Vickers hardness was measured at various distances of a cross section of the TIG joints of JLF-1 steel according to the heating history of each position. Finally, the fracture surface was observed by scanning electron microscopy (SEM).

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Potential of polylactic-co-glycolic acid (PLGA) for delivery Jembrana disease DNA vaccine Model (pEGFP-C1-tat)

  • Unsunnidhal, Lalu;Wasito, Raden;Setyawan, Erif Maha Nugraha;Warsani, Ziana;Kusumawati, Asmarani
    • Journal of Veterinary Science
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    • v.22 no.6
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    • pp.76.1-76.15
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    • 2021
  • Background: The development of a vaccine for Jembrana disease is needed to prevent losses in Indonesia's Bali cattle industry. A DNA vaccine model (pEGFP-C1-tat) that requires a functional delivery system will be developed. Polylactic-co-glycolic acid (PLGA) may have potential as a delivery system for the vaccine model. Objectives: This study aims to evaluate the in vitro potential of PLGA as a delivery system for pEGFP-C1-tat. Methods: Consensus and codon optimization for the tat gene was completed using a bioinformatic method, and the product was inserted into a pEGFP-C1 vector. Cloning of the pEGFP-C1-tat was successfully performed, and polymerase chain reaction (PCR) and restriction analysis confirmed DNA isolation. PLGA-pEGFP-C1-tat solutions were prepared for encapsulated formulation testing, physicochemical characterization, stability testing with DNase I, and cytotoxicity testing. The PLGA-pEGFP-C1-tat solutions were transfected in HeLa cells, and gene expression was observed by fluorescent microscopy and real-time PCR. Results: The successful acquisition of transformant bacteria was confirmed by PCR. The PLGA:DNA:polyvinyl alcohol ratio formulation with optimal encapsulation was 4%:0.5%:2%, physicochemical characterization of PLGA revealed a polydispersity index value of 0.246, a particle size of 925 nm, and a zeta potential value of -2.31 mV. PLGA succeeded in protecting pEGFP-C1-tat from enzymatic degradation, and the percentage viability from the cytotoxicity test of PLGA-pEGFP-C1-tat was 98.03%. The PLGA-pEGFP-C1-tat demonstrated luminescence of the EGFP-tat fusion protein and mRNA transcription was detected. Conclusions: PLGA has good potential as a delivery system for pEGFP-C1-tat.

A Role for Peroxidasin PXN-1 in Aspects of C. elegans Development

  • Lee, Juyeon;Bandyopadhyay, Jaya;Lee, Jin Il;Cho, Injeong;Park, Daeho;Cho, Jeong Hoon
    • Molecules and Cells
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    • v.38 no.1
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    • pp.51-57
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    • 2015
  • The Caenorhabditis elegans peroxidasins, PXN-1 and PXN-2, are extracellular peroxidases; pxn-2 is involved in muscle-epidermal attachment during embryonic morphogenesis and in specific axon guidance. Here we investigate potential roles of the other homologue of peroxidasin, pxn-1, in C. elegans. A pxn-1 deletion mutant showed high lethality under heat-stress conditions. Using a transcriptional GFP reporter, pxn-1 expression was observed in various tissues including neurons, muscles, and hypodermis. A translational fusion showed that PXN-1::GFP was secreted and localized in extracellular matrix, particularly along body wall muscles and pharyngeal muscles. Various neuronal developmental defects were observed in pxn-1 mutants and in pxn-1 over-expressing animals, including handedness, branching, breakage, tangling, and defasciculation. These results suggest that pxn-1, like other peroxidasins, plays an important role throughout development.

A Numerical Study on the Performance Analysis of a Solar Air Heating System with Forced Circulation Method (강제순환 방식의 공기가열식 태양열 집열기의 성능분석에 관한 수치해석 연구)

  • Park, Hyeong-Su;Kim, Chul-Ho
    • Journal of the Korea Academia-Industrial cooperation Society
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    • v.18 no.3
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    • pp.122-126
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    • 2017
  • The aim of this study was to develop a device for solving the heating problem of living space using heated air, utilizing a simple air heater type collector for solar energy. At the present time, this study assessed the possibility of a development system through theoretical calculations for the amount of available energy according to the size change of the air-heated solar energy collector. To produce and supply hot water using the heat energy of the sun, hot water at $100^{\circ}C$ or less was produced using a flat or vacuum tube type collector. The purpose of this study was to research the air heating type solar collector that utilizes heating energy with heating air above $75^{\circ}C$, by designing and manufacturing an air piping type solar collector that is a simpler type than a conventional solar collector system. The analysis results were obtained for the generated air temperature ($^{\circ}C$) and the production of air (kg/h) to determine the performance of air heating by an air-heated solar collector according to the heat transfer characteristics in the collector of the model when a specified amount of heat flux was dropped into a solar collector of a certain size using PHOENICS, which is a heat flow analysis program applying the Finite Volume Method. From the analysis result, the temperature of the air obtained was approximately $40.5^{\circ}C$, which could be heated using an air heating tube with an inner diameter of 0.1m made of aluminum in a collector with a size of $1.2m{\times}1.1m{\times}0.19m$. The production of air was approximately 161 m3/h. This device can be applied to maintain a suitable environment for human activity using the heat energy of the sun.

Sequencing of cDNA Clones Expressed in Adipose Tissues of Korean Cattle

  • Bong, J.J.;Tong, K.;Cho, K.K.;Baik, M.G.
    • Asian-Australasian Journal of Animal Sciences
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    • v.18 no.4
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    • pp.483-489
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    • 2005
  • To understand the molecular mechanisms that regulate intramuscular fat deposition and its release, cDNA clones expressed in adipose tissues of Korean cattle were identified by differential screening from adipose tissue cDNA library. By partial nucleotide sequencing of 486 clones and a search for sequence similarity in NCBI nucleotide databases, 245 clones revealed unique clones. By a functional grouping of the clones, 14% of the clones were categorized to metabolism and enzyme-related group (stearoyl CoA desaturase, lactate dehydrogenase, fatty acid synthase, ATP citrate lyase, lipoprotein lipase, acetyl CoA synthetase, etc), and 6% to signal transduction/cell cycle-related group (C/EBP, cAMP-regulated phosphoprotein, calmodulin, cyclin G1, cyclin H, etc), and 4% to cytoskeleton and extracellular matrix components (vimentin, ankyrin 2, gelosin, syntenin, talin, prefoldin 5). The obtained 245 clones will be useful to study lipid metabolism and signal transduction pathway in adipose tissues and to study obesity in human. Some clones were subjected to full-sequencing containing open reading frame. The cDNA clone of bovine homolog of human prefoldin 5 gene had a total length of 959 nucleotides coding for 139 amino acids. Comparison of the deduced amino acid sequences of bovine prefoldin 5 with those of human and mouse showed over 95% identity. The cDNA clone of bovine homolog of human ubiquitin-like/S30 ribosomal fusion protein gene had a total length of 484 nucleotides coding for 133 amino acids. Comparison of the deduced amino acid sequences of bovine ubiquitin-like/S30 ribosomal fusion protein gene with those of human, rat and mouse showed over 97% identity. The cDNA clone of bovine homolog of human proteolipid protein 2 mRNA had a total length of 928 nucleotides coding for 152 amino acids. Comparison of the deduced amino acid sequences of bovine proteolipid protein 2 with those of human and mouse showed 87.5% similarity. The cDNA clone of bovine homolog of rat thymosin beta 4 had a total length of 602 nucleotides coding for 44 amino acids. Comparison of the deduced amino acid sequences of bovine thymosin beta 4 gene with those of human, mouse and rat showed 93.1% similarity. The cDNA clone of bovine homolog of human myotrophin mRNA had a total length of 790 nucleotides coding for 118 amino acids. Comparison of the deduced amino acid sequences of bovine myotrophin gene with those of human, mouse and rat showed 83.9% similarity. The functional role of these clones in adipose tissues needs to be established.

Properties of AlN epilayer grown on 6H-SiC substrate by mixed-source HVPE method (6H-SiC 기판 위에 혼합소스 HVPE 방법으로 성장된 AlN 에피층 특성)

  • Park, Jung Hyun;Kim, Kyoung Hwa;Jeon, Injun;Ahn, Hyung Soo;Yang, Min;Yi, Sam Nyung;Cho, Chae Ryong;Kim, Suck-Whan
    • Journal of the Korean Crystal Growth and Crystal Technology
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    • v.30 no.3
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    • pp.96-102
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    • 2020
  • In this paper, AlN epilayers on 6H-SiC (0001) substrate are grown by mixed source hydride vapor phase epitaxy (MS-HVPE). AlN epilayer of 0.5 ㎛ thickness was obtained with a growth rate of 5 nm per hour. The surface of AlN epilayer grown on 6H-SiC (0001) substrate was investigated by field emission scanning electron microscopy (FE-SEM) and energy dispersive X-ray spectroscopy (EDS). Dislocation density was considered through HR-XRD and related calculations. A fine crystalline AlN epilayer with screw dislocation density of 1.4 × 109 cm-2 and edge dislocation density of 3.8 × 109 cm-2 was confirmed. The AlN epilayer on 6H-SiC (0001) substrate grown by using the mixed source HVPE method could be applied to power devices.

Nucleo-cytoplasmic Interactions of Bovine Oocytes and Embryos Following Nuclear Transplantation (핵이식에 의한 소 난자 및 초기배의 핵-세포질의 상호작용에 관한 연구)

  • 김정익;양부근;정희태
    • Korean Journal of Animal Reproduction
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    • v.17 no.4
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    • pp.287-294
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    • 1994
  • This study was to investigate the effects of electrofusion, activation and developmental stage of donor embryos on in vitro development of nuclear transplant bovine embryos. A single blastomere nucleus from 8-cell to morula stage embryos produced by in vitro fertilization(IVF) was transferred into a recipient oocyte enucleated at 23∼25 h after in vitro maturation(IVM) or into a recipient oocyte enucleated and cultured for 14∼15 h. In one experiment the nuclear transplant embryos were subjected to additional activation treatments. Fusion rate of nuclear transplant eggs was high at direct current(D.C) voltages of 1.0 and 1.5 kV/cm 991.5 and 93.3%, respectively), but decreased at 2.0kV/cm (81.8%). Additional activation treatments by electric pulases or 7% ethanol did not affect the cleavage and development of nuclear transplant embryos. Development of nuclear transplant embryos slightly increased by delayed nuclear transfer and fusion (42∼43 h after IVM). With this system, blastocysts were obtained from transfer of 8-cell to morula stage donor nuclei (9.6%∼2.4%). The result of this study suggests that nucleo-cytoplasmic interactins, expecially activation of ooplast are very important for the development of nuclear transplant embryos, and donor cell stage does not affect the development of nuclear transplant embryos.

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The Clinical Experience of Computed Tomographic-Guided Navigation System in C1-2 Spine Instrumentation Surgery

  • Kim, Sang-Uk;Roh, Byoung-Il;Kim, Seong-Joon;Kim, Sang-Don
    • Journal of Korean Neurosurgical Society
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    • v.56 no.4
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    • pp.330-333
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    • 2014
  • Objective : To identify the accuracy and efficiency of the computed tomographic (CT)-based navigation system on upper cervical instrumentation, particularly C1 lateral mass and C2 pedicle screw fixation compared to previous reports. Methods : Between May 2005 and March 2014, 25 patients underwent upper cervical instrumentation via a CT-based navigation system. Seven patients were excluded, while 18 patients were involved. There were 13 males and five females; resulting in four degenerative cervical diseases and 14 trauma cases. A CT-based navigation system and lateral fluoroscopy were used during the screw instrumentation procedure. Among the 58 screws inserted as C1-2 screws fixation, their precise positions were evaluated by postoperative CT scans and classified into three categories : in-pedicle, non-critical breach, and critical breach. Results : Postoperatively, the precise positions of the C1-2 screws fixation were 81.1% (47/58), and 8.6% (5/58) were of non-critical breach, while 10.3% (6/58) were of critical breach. Most (5/6, 83.3%) of the critical breaches and all of non-critical breaches were observed in the C2 pedicle screws and there was only one case of a critical breach among the C1 lateral mass screws. There were three complications (two vertebral artery occlusions and a deep wound infection), but no postoperative instrument-related neurological deteriorations were seen, even in the critical breach cases. Conclusion : Although CT-based navigation systems can result in a more precise procedure, there are still some problems at the upper cervical spine levels, where the anatomy is highly variable. Even though there were no catastrophic complications, more experience are needed for safer procedure.