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Cytotoxic T Lymphocytes Elicited by Dendritic Cell-Targeted Delivery of Human Papillomavirus Type-16 E6/E7 Fusion Gene Exert Lethal Effects on CaSki Cells

  • Wu, Xiang-Mei;Liu, Xing;Jiao, Qing-Fang;Fu, Shao-Yue;Bu, You-Quan;Song, Fang-Zhou;Yi, Fa-Ping
    • Asian Pacific Journal of Cancer Prevention
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    • 제15권6호
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    • pp.2447-2451
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    • 2014
  • Human papillomavirus (HPV) is the primary etiologic agent of cervical cancer. Consideration of safety and non human leukocyte antigen restriction, protein vaccine has become the most likely form of HPV therapeutic vaccine, although none have so far been reported as effective. Since tumor cells consistently express the two proteins E6 and E7, most therapeutic vaccines target one or both of them. In this study, we fabricated DC vaccines by transducing replication-defective recombinant adenoviruses expressing E6/E7 fusion gene of HPV-16, to investigate the lethal effects of specific cytotoxic T lymphocytes (CTL) against CaSki cells in vitro. Mouse immature dendritic cells (DC) were generated from bone marrow, and transfected with pAd-E6/E7 to prepare a DC vaccine and to induce specific CTL. The surface expression of CD40, CD68, MHC II and CD11c was assessed by flow cytometry (FCM), and the lethal effects of CTL against CaSki cells were determined by DAPI, FCM and CCK-8 methods. Immature mouse DC was successfully transfected by pAd-E6/E7 in vitro, and the transfecting efficiency was 40%-50%. A DC vaccine was successfully prepared and was used to induce specific CTL. Experimental results showed that the percentage of apoptosis and killing rate of CaSki cells were significantly increased by coculturing with the specific CTL (p <0.05). These results illustrated that a DC vaccine modified by HPV-16 E6/E7 gene can induce apoptosis of CaSki cells by inducing CTL, which may be used as a new strategy for biological treatment of cervical cancer.

자동차의 배기관 VOCs 배출 특성 (Exhaust VOCs Emission Characteristics from Motor Vehicles)

  • 유영숙;류정호;한종수;김선문;임철수;김대욱;이동민;이중구;엄명도;김종춘
    • 한국대기환경학회지
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    • 제24권3호
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    • pp.275-283
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    • 2008
  • Since mobile source is a major source of VOCs, quantifying emissions from motor vehicles is an important factor to control VOCs in atmosphere. In this study, in order to evaluate tailpipe VOCs emissions from motor vehicles, mass emissions of non-methane volatile organic compounds from 45 vehicles were determined. Measurements were made on a chassis dynamometer using CVS-75 mode and speed specific drive modes. Target VOCs are 53 compounds determined as the volatile ozone precursors. The individual VOCs composition of vehicle emission and emission rates were also determined. In case of gasoline vehicles, VOCs emission from over 80,000 km vehicles were about 46% larger than less 80,000 km vehicles. The difference in benzene and toluene according to driving mileage was 44% and 26% respectively. The composition of VOCs were different by fuel type. The order of VOCs composition was paraffins>aromatics>olefins in gasoline vehicle emissions, paraffins>olefins>aromatics in light duty diesel vehicle emissions. The VOCs emissions were decreased as vehicle speed increasing. These results will be used to calculate total VOCs emissions from automobiles in the future.

간질아동에게 제공된 심리사회적 중재의 효과에 대한 연구동향: Systematic Review (Trends of Research on the Effects of Psychosocial Intervention for Children with Epilepsy: Systematic Review)

  • 유하나;윤오복
    • 한국콘텐츠학회논문지
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    • 제14권6호
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    • pp.219-228
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    • 2014
  • 목적 본 연구는 간질 아동에게 제공된 심리사회적 중재들의 효과에 대한 연구동향을 밝혀 추후 간질아동을 위한 간호중재의 기초자료로 활용하기 위함이다. 방법 PRISMA 문헌선택 흐름도에 따라 데이터베이스(Ovid-MEDLINE, EMBASE, PubMed, CINAHL, The Cochrane library, KMbase, and Koreamed)를 통해 454개의 논문들이 검색되었고 선정 및 제외기준에 의해 7개의 논문이 최종적으로 선택되었다. 2명의 연구자가 분석에 포함된 모든 연구를 독립적으로 검토하였고, 방법론적인 질 평가를 위해 코크란의 Risk of Bias 도구와 국내의 Risk of Bias Assessment tool for Non-randomized Study 도구를 이용하였다. 결과 중재 전략의 형태는 4가지로 분류되었다: 교육; 상담; 인지행동치료; 신체활동. 중재의 목표들은 매우 다양하였다. 결론 각각의 중재들은 간질관련 건강 결과에 부분적으로 긍정적인 효과를 보여주었지만 불충분한 중재 정보, 타당도가 떨어지는 결과 측정법, 적은 표본 수, 연구 설계의 제한점과 같은 방법론적인 약점들이 있었다.

도시텃밭 운영 프로그램이 참여자 공동체의식에 미치는 영향 (Effects of the Urban Farm Program on the Participants' Sense of Community)

  • 박태호;이인성
    • 한국조경학회지
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    • 제40권5호
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    • pp.119-128
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    • 2012
  • 본 연구에서는 도시텃밭 프로그램이 공동체의식에 미치는 효과를 규명하고자 서울의 3개 공공도시텃밭을 선정하여 설문조사를 실시하였다. 연구결과는 다음과 같다. 첫째, 텃밭프로그램이 참여자의 상호교제와 공동체의식에 미치는 영향이 다각도로 확인되었다. 지속적 프로그램이 있는 텃밭과 그렇지 못한 텃밭 사이에는 내부교제와 공동체의식에서 차이가 있었으며, 프로그램참여자가 비참여자에 비해 높은 수준의 텃밭활동, 내 외부교제 및 공동체의식을 보였고, 경로분석에서 도 프로그램참여가 공동체의식에 가장 큰 영향을 미치는 변수로 나타났다. 둘째, 자가점유, 거주기간, 참여인원이 공동체의식에 영향을 미치는데, 이는 거주기간이 긴 자가점유자, 특히 1인 및 노인 가구 등 사회적 소외계층을 대상으로 텃밭프로그램이 운영되어야 함을 시사한다. 셋째, 농업교육보다 지역사회와 연계된 다양한 문화 프로그램의 만족도가 높았다. 이 결과들은 도시텃밭의 공동체의식 형성효과를 높이기 위해서는 지속적이고 다양한 프로그램이 필요함을 보여준다.

Application of LATE-PCR to Detect Candida and Aspergillus Fungal Pathogens by a DNA Hybridization Assay

  • Gopal, Dhayaalini Bala;Lim, Chua Ang;Khaithir, Tzar Mohd Nizam;Santhanam, Jacinta
    • 한국미생물·생명공학회지
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    • 제45권4호
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    • pp.358-364
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    • 2017
  • Asymmetric PCR preferentially amplifies one DNA strand for use in DNA hybridization studies. Linear-After-The-Exponential-PCR (LATE-PCR) is an advanced asymmetric PCR method which uses innovatively designed primers at different concentrations. This study aimed to optimise LATE-PCR parameters to produce single-stranded DNA of Candida spp. and Aspergillus spp. for detection via probe hybridisation. The internal transcribed spacer (ITS) region was used to design limiting primer and excess primer for LATE-PCR. Primer annealing and melting temperature, difference of melting temperature between limiting and excess primer and concentration of primers were optimized. In order to confirm the presence of single-stranded DNA, the LATE-PCR product was hybridised with digoxigenin labeled complementary oligonucleotide probe specific for each fungal genus and detected using anti-digoxigenin antibody by dot blotting. Important parameters that determine the production of single-stranded DNA in a LATE-PCR reaction are difference of melting temperature between the limiting and excess primer of at least $5^{\circ}C$ and primer concentration ratio of excess primer to limiting primer at 20:1. LATE-PCR products of Candida albicans, Candida parapsilosis, Candida tropicalis and Aspergillus terreus at up to 1:100 dilution and after 1 h hybridization time, successfully hybridised to respective oligonucleotide probes with no cross reactivity observed between each fungal genus probe and non-target products. For Aspergillus fumigatus, LATE-PCR products were detected at 1:10 dilution and after overnight hybridisation. These results indicate high detection sensitivity for single-stranded DNA produced by LATE-PCR. In conclusion, this advancement of PCR may be utilised to detect fungal pathogens which can aid the diagnosis of invasive fungal disease.

Combination of Doxorubicin with Gemcitabine-Incorporated G-Quadruplex Aptamer Showed Synergistic and Selective Anticancer Effect in Breast Cancer Cells

  • Joshi, Mili;Choi, Jong-Soo;Park, Jae-Won;Doh, Kyung-Oh
    • Journal of Microbiology and Biotechnology
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    • 제29권11호
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    • pp.1799-1805
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    • 2019
  • Doxorubicin (DOX) is one of the most effective anticancer agents used for the treatment of multiple cancers; however, its use is limited by its short half-life and adverse drug reactions, especially cardiotoxicity. In this study, we found that the conjugate of DOX with APTA12 (Gemcitabine incorporated G-quadruplex aptamer) was significantly more cancer selective and cytotoxic than DOX. The conjugate had an affinity for nucleolin, with higher uptake and retention into the cancer cells than those of DOX. Further, it was localized to the nucleus, which is the target site of DOX. Owing to its mechanism of action, DOX has the ability to intercalate into the nucleotides thus making it a suitable drug to form a conjugate with cancer selective aptamers such as APTA12. The conjugation can lead to selectively accumulate in the cancer cells thus decreasing its potential nonspecific as well as cardiotoxic side effects. The aim of this study was to prepare a conjugate of DOX with APTA12 and assess the chemotherapeutic properties of the conjugate specific to cancer cells. The DOX-APTA12 conjugate was prepared by incubation and its cytotoxicity in MCF-10A (non-cancerous mammary cells) and MDA-MB-231 (breast cancer cells) was assessed. The results indicate that DOX-APTA12 conjugate is a potential option for chemotherapy especially for nucleolin expressing breast cancer with reduced doxorubicin associated side effects.

Effect of Histone Deacetylase Inhibitors on Differentiation of Human Bone Marrow-derived Stem Cells Into Neuron-like Cells

  • Jang, Sujeong;Park, Seokho;Cho, Hyong-Ho;Yang, Ung;Kang, Maru;Park, Jong-Seong;Park, Sah-Hoon;Jeong, Han-Seong
    • 통합자연과학논문집
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    • 제12권4호
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    • pp.133-141
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    • 2019
  • Mesenchymal stem cells (MSCs) are known to differentiate into multiple lineages, making neurogenic differentiation an important target in the clinical field. In the present study, we induced the neurogenic differentiation of cells using histone deacetylase (HDAC) inhibitors and studied their mechanisms for further differentiation in vitro. We treated cells with the HDAC inhibitors, MS-275 and NaB; and found that the cells had neuron-like features such as distinct bipolar or multipolar morphologies with branched processes. The mRNA expressions encoding for NEFL, MAP2, TUJ1, OLIG2, and SYT was significantly increased following HDAC inhibitors treatment compared to without HDAC inhibitors; high protein levels of MAP2 and Tuj1 were detected by immunofluorescence staining. We examined the mechanisms of differentiation and found that the Wnt signaling pathway and downstream mitogen-activate protein kinase were involved in neurogenic differentiation of MSCs. Importantly, Wnt4, Wnt5a/b, and Wnt11 protein levels were highly increased after treatment with NaB; signals were activated through the regulation of Dvl2 and Dvl3. Interestingly, NaB treatment increased the levels of JNK and upregulated JNK phosphorylation. After MS-275 treatment, Wnt protein levels were decreased and GSK-3β was phosphorylated. In this cell, HDAC inhibitors controlled the non-canonical Wnt expression by activating JNK phosphorylation and the canonical Wnt signaling by targeting GSK-3β.

비정질 금속 기판상에 증착된 YBCO 박막의 결정성에 대한 CEO$_2$ 완충막의 효과 (Effect of CeO$_2$ buffer layer on the crystallization of YBCO thin film on Hastelloy substrate)

  • 김성민;이상렬
    • 한국초전도학회:학술대회논문집
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    • 한국초전도학회 1999년도 High Temperature Superconductivity Vol.IX
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    • pp.392-396
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    • 1999
  • Superconducting YBa$_2Cu_3O_{7-{\delta}}$(YBCO) thin films were grown on Hastelloy(Ni-Cr-Mo alloys) with CeO$_2$ buffer layer in-situ by pulsed laser deposition in a multi-target processing chamber. To apply superconducting property on power transmission line, we have deposited YBCO thin film on flexible metallic substrate. However, it is difficult to grow the YBCO films on flexible metallic substrates due to both interdiffusion problem between metallic substrate and superconducting overlayers and non-crystallization of YBCO on amorphous substrate. It is necessary to use a buffer layer to overcome the difficulties. We have chosen CeO$_2$ as a buffer layer which has cubic structure of 5.41 ${\AA}$ lattice parameter and only 0.2% of lattice mismatch with 3.82 ${\AA}$ of a-axis lattice parameter of YBCO on [110] direction of CeO$_2$ In order to enhance the crystallization of YBCO films on metallic substrates, we deposited CeO$_2$ buffer layers with varying temperature and 02 pressure. By XRD, it is observed that dominated film orientation is strongly depending on the deposition temperature of CeO$_2$ layer. The dominated orientation of CeO$_2$ buffer layer is changed from (200) to(111) by increasing the deposition temperature and this transition affects the crystallization of YBCO superconducting film on CeO$_2$ buffered Hastelloy.

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RF Magnetron Sputtering공정에 의해 IT유리에 적층시킨 Silicon Nitride 박막의 특성 (Characteristics of Silicon Nitride Deposited Thin Films on IT Glass by RF Magnetron Sputtering Process)

  • 손정일;김광수
    • 한국재료학회지
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    • 제30권4호
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    • pp.169-175
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    • 2020
  • Silicon nitride thin films are deposited by RF (13.57 MHz) magnetron sputtering process using a Si (99.999 %) target and with different ratios of Ar/N2 sputtering gas mixture. Corning G type glass is used as substrate. The vacuum atmosphere, RF source power, deposit time and temperature of substrate of the sputtering process are maintained consistently at 2 ~ 3 × 10-3 torr, 30 sccm, 100 watt, 20 min. and room temperature, respectively. Cross sectional views and surface morphology of the deposited thin films are observed by field emission scanning electron microscope, atomic force microscope and X-ray photoelectron spectroscopy. The hardness values are determined by nano-indentation measurement. The thickness of the deposited films is approximately within the range of 88 nm ~ 200 nm. As the amount of N2 gas in the Ar:N2 gas mixture increases, the thickness of the films decreases. AFM observation reveals that film deposited at high Ar:N2 gas ratio and large amount of N2 gas has a very irregular surface morphology, even though it has a low RMS value. The hardness value of the deposited films made with ratio of Ar:N2=9:1 display the highest value. The XPS spectrum indicates that the deposited film is assigned to non-stoichiometric silicon nitride and the transmittance of the glass with deposited SiO2-SixNy thin film is satisfactory at 97 %.

Properties of a New Adhesive Composed of Gambir-Sucrose

  • SUCIPTO, Tito;WIDYORINI, Ragil;PRAYITNO, Tibertius Agus;LUKMANDARU, Ganis
    • Journal of the Korean Wood Science and Technology
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    • 제48권3호
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    • pp.303-314
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    • 2020
  • Gambir is a non-wood forest product with a potential of being used as wood adhesive, due to about 33% catechin in it. Meanwhile, catechins and sucrose have not been studied as adhesives. Therefore, basic characteristics of gambir-sucrose adhesives were investigated. In this research, adhesives were prepared by dissolving gambir and sucrose in distilled water, at different blending ratios of the gambir/sucrose such as 100/0, 75/25, 50/50, and 25/75 wt%. Furthermore, gas chromatography-mass spectrometry (GC-MS) was employed to determine the gambir chemical compositions, and Fourier transform-infrared (FTIR) spectroscopy was carried out to identify chemical bonds. Particleboards with a target density of 0.8 g/㎤ were then manufactured by hot-pressing for 10 min at 200℃. The internal bond (IB) strength of particleboard was subsequently measured. Based on the GC-MS analysis, 31.11% of catechin was identified. In addition, the viscosity, density, solid content, and gelation time of the adhesives, and insoluble matter content (IMC) in boiling water were 7.30~33.24 mPa.s, 1.2~1.3 g/㎤, 25.56~28.44%, 73~420 min, and 29.75~62.10%, respectively. Adding sucrose to the adhesive was observed to raise the IMC from 49.05 to 62.10%, at 180℃ and 200℃. FT-IR analysis showed that the gambir absorption peaks occurred at approximately 1620 cm-1, assigned to the C=O stretching of 5-hydroxymethylfurfural, which tended to increase with the addition of sucrose. The reaction between gambir and sucrose was observed in the form of the dimethylene ether bridge. The 25/75 wt% gambir-sucrose adhesives and 200℃ hot-pressed temperature resulted in the highest IB strength (0.89 MPa), and met the requirement of JIS A5908-2003 type 18. Consequently, the gambir-sucrose adhesive could be used as a particleboard adhesive.