저독성 키토산 금 나노입자를 이용한 shRNA의 폐암세포에서 전달

정새로미 崇實大學校 2012 국내석사

본 연구에서는 독성이 낮은 금 나노입자를 만들고, small hairpin RNA (shRNA)를 붙여서 세포내에서의 uptake를 dark-field microscopy (DFM) 와 공초점 라만 분광기를 통해 (confocal Raman spectroscopy) 관찰하는 것을 연구하였다. 이들의 결합반응은 (-)전하를 띠는 shRNA 를 (+) 전하를 띠는 키토산 금 나노입자 표면위에 정전기적 상호작용을 통하여 일어나고 infrared spectroscopy (IR)과 visible absorption 으로 확인을 하였다. (+) 전하를 띠는 키토산 금 나노입자를 화학적 환원방법을 통하여 준비를 하고, A549 세포내로 들어간 나노입자를 TEM (tranmission electron microscopy) 사진으로부터 알 수 있었다. 금 나노입자에 shRNA가 bioconjugates 된 후 세포 내에 위치는 공초점 라만 분광기를 통해 관찰 할 수 있었다. 키토산 금 나노입자는 좋은 gene silencing효과를 나타낼 수 있었고 널리 쓰이는 BPEI (branched ,polyethylenimine) 보다 더 적은 독성을 나타낸다는 걸 확인 할 수 있었다. The research for the present study focused on the preparation of low-toxicity gold nanoparticles (Au NPs) attached with small hairpin RNA (shRNA) and on monitoring their intracellular distribution by means of dark-field microscopy (DFM) and confocal Raman spectroscopy. The conjugation reactions of negatively charged shRNA onto positively charged chitosan Au NPs should occur via an electrostatic interaction as examined by visible absorption and infrared spectroscopy. Positively charged chitosan gold nanoparticles (Au NPs) prepared by chemical reduction were found well-internalized inside human lung adenocarcinoma A549 cells from the transmission electron microscopy (TEM) images. We determined the localization of internalized Au NP-shRNA bioconjugates and found that Au NPs can be monitored inside a single cell using a confocal Raman spectroscopic tool. Chitosan Au NPs appeared to show good gene silencing efficiency with less toxicity than widely used branched, polyethyleneimine (BPEI).

또래와의 사회적기술지향 음악놀이활동이 자폐성 아동의 사회적 상호작용에 미치는 효과

정새로미 공주대학교 대학원 2003 국내석사

국문초록 또래와의 사회적 기술 지향 음악놀이활동이 자폐성 아동의 사회적 상호작용에 미치는 효과 공주대학교 대학원 특수교육학과 정서장애교육전공 정새로미 본 연구의 목적은 또래와의 사회적 기술 지향 음악놀이활동이 두 명의 자폐성 아동의 사회적 상호작용에 어떠한 영향을 미치는지를 알아보고자 하였다. 실험 방법은 단일대상연구의 반전 설계법을 사용하였으며, 전체 총 30회기에 걸쳐 기초선(5회기), 중재 1(8회기), 반전(5회기), 중재 2(8회기), 유지(4회기)를 실시하였다. 실험은 전 기간 동안 1주일에 3일(월, 수, 금) 동안 총 40분씩 자유놀이와 음악놀이활동이 진행되었으며, 사회적 상호작용의 발생률은 자유놀이 시간 15분중 10분을 비디오로 촬영하여 15초 간격의 등간기록법으로 부분간격기록하여 각 대상학생들의 사회적 상호작용 발생률을 분석하였다. 이 연구의 결과를 종합하여 결론을 내리면 다음과 같다. 첫째, 또래와의 사회적 기술 지향 음악놀이활동은 자폐성 아동의 사회적 상호작용 습득에 효과적이었다. 둘째, 또래와의 사회적 기술 지향 음악놀이활동을 통해 습득된 사회적 상호작용은 중재가 종결된 후 유지 검사에서도 자폐성 아동에게 그 효과가 있었다. 이러한 결과는 또래와의 사회적 기술 지향 음악놀이 활동이 경증-중간 자폐성 아동인 아동 A에게는 목표행동이었던 주의집중, 모방, 관심공유에 있어서 전반적으로 골고루 증가하는 경향을 나타내어 적절한 중재전략이었다고 볼 수 있었다. 그러나 중증의 자폐성 아동인 아동 B에게는 또래와의 사회적 기술 지향 음악놀이활동이 목표행동이었던 주의집중과 모방에는 적절한 중재전략이었으나 관심공유에 있어서는 그다지 효과적이지 못했다고 볼 수 있다. 그러므로 중증의 자폐성 아동에게는 통합된 환경 안에서 보다 심화되고 단계적인 중재 전략이 필요하며 그에 따른 지속적인 연구가 이루어져야 함을 시사하고 있다. 이상의 연구 결과들을 종합하여 결론을 맺으면 다음과 같다. 또래와의 사회적 기술 지향 음악놀이활동은 자폐성 아동의 사회적 상호작용 의 습득에 있어서 그 효과가 있었으며, 하위 행동으로 본 주의집중, 모방, 관심공유에 있어서 경증-중간 자폐성 아동은 주의집중, 모방, 관심공유가 모두 골고루 증가하는 양상을 보였다. 그러나 중증의 자폐성 아동은 주의집중이나 모방은 변화를 보였으나 관심공유의 경우에는 어려운 것으로 나타났다. 2주 후 실시된 유지 검사에서 두 아동들은 모두 습득된 사회적 상호작용이 유지되고 있는 것으로 나타났으며, 하위 행동에 있어서도 경증-중간 자폐 아동은 주의집중, 모방, 관심공유 행동이 모두 유지되었으나, 중증 자폐 아동은 주의집중, 모방만 유지되고 관심공유 행동은 전혀 나타나지 않아 후속연구의 필요성과 적합한 프로그램의 개발을 시사하고 있다. ABSTRACT The effects of social skill-oriented music play activities with peers on social interaction of autistic children Sae Ro Mi Jeong Department of Special Education Graduate School, Kong Ju National University, Kong Ju, Korea (Supervised by Professor Seong Hee Han, Ph.D.) The purpose of this study was to investigate the effects of social skill- oriented music play activities with peers on social interaction of autistic children. For this purpose, two children with autism and twelves nonhandicapped peers were selected from one social welfare center. In this studies, reversal design method was used to identify the effect of interagency and an experiment was progressed a long a baseline five times, the first interagency step eight times, reverse five times, the second interagency step eight times, maintenance four times. In processing the test, social skill-based music play activities have been carried out to subjects for 20 minutes, and the interval recording has been used before each activity for 10 minutes to collect data. The results obtained from this study are summarized as the following: First, social skill-oriented music play activities with peers had an effect on improving social interaction of autistic children. Second, social skill-oriented music play activities with peers had an effect on maintaining the improved social interaction of autistic children.

≪聊齋志異≫의 愛情故事에 나타난 男性의 自己實現 硏究

정새로미 공주대학교 교육대학원 2010 국내석사

明末清初的蒲松龄科举失败, 又适逢王朝更迭, 社会飘摇之际, 个人的境遇与国家的遭遇构成了明清文人独特的人格特征. 从而, 在这现象之后, 折射出的是封建社会的衰微与没落以及文人内心的苦闷和悲凉, ≪聊齋志異≫表達了作家蒲松齡的思想, 愛憎感情. ≪聊齋志異≫篇目四百九十二篇之中, 情愛篇章有九十七篇, 約為五分之一, 其中有许多乃人與異類之情愛故事. 这些作品的女性命运不同, 形态各异的女性大致, 逐步變異至≪聊齋志異≫中令人嚮往的女性, 可說是男性普遍欲求下的共同心理投射, 也正與榮格所說「無意識」與「原型理論」相契合. 本論文嘗試利用榮格心理學中的無意識, 阿尼瑪, 人格面具, 阴影等理論進行狐女形象分析, 期望能從心理學的角度, 對於以男性心理所創造出的女性形象, 有較完整的瞭解. 本論文嘗試透過榮格的心理學進行分析, 試圖為文中眾多的女性形象找尋其共有的創作基礎. 即蒲松齡在女性形象敘寫背後所隱含的心理因素, 期能對長期以來令人著迷的作品有深一層的理解.

Electrochemical biosensor for Chemokine ligand and CXCR2 receptor affinity

정새로미 부산대학교 2012 국내석사

Chemokine Ligands bind to its cognate receptor CXCR2 to induce inflammatory responses, wound healing, tumorogenesis, and neuronal survival. CXCR2 is the receptor for CXCL5 and CXCL8 involved in angiogenic CXC chemokine-mediated angiogenesis. CXCR2 is promiscuous and binds multiple chemokine ligand with high affinity, including CXCL5, CXCL8. Chemokine ligands and receptors have been shown to play important roles in mediating non-small cell lung cancer-associated angiogenesis and organ-specific metastases. Compared with CXCL8, CXCL5 was reported to have a higher degree of correlation with NSCLC-derived angiogenesis. Conducting polymer was polymerized on GCE before immobilizing CXCR2 to fabricate sensor probe. CXCL5 was allowed to react with the sensing probe. The sensor probe was characterized using Cyclic voltammetry, Impedance and QCM. Various parameters such as pH, temperature and ligands concentration were optimized.

Development of New Approaches for Electrochemical Detection of Clinically Relevant Molecules

정새로미 부산대학교 대학원 2019 국내박사

Electrochemical platforms such as biosensors and microfluidic channels for clinically relevant molecules detection have been developed towards simplify detection process and miniaturized device. Especially, electrochemical biosensors can be easily applied to hand-held device for clinical monitoring due to its coherent characteristics in terms of easy-to-use, simple, fast, selective, and sensitive properties. In addition, microfluidic channels have been considered to be very promising devices to generate new recognition molecules which can be applied to biosensors. These platforms make quantitative and qualitative analysis possible based on interaction between analytes and recognition molecules such as antibodies, natural receptors, and aptamers. Recently, it has been received attention as a point-of-care testing device. For this, it is essential to develop a device that can measure accurately and accurately with high sensitivity, as well as to discover a recognition molecule. To achieve this, clinical molecules such as chemokine, thrombin, rocuronium were electrochemically detected using recognition molecules and electrode catalysts and aptamers for malaria biomarker was generated using AC potential modulated microfluidic channels. In chapter 1, general information of electrochemical biosensors and AC applied microfluidic channel using potentiometric, amperometric, and conductometric method, is introduced. To apply those electrochemical devices to various field such as biosensor, chemical sensor, and immunosensor, recognition molecules and electrode catalysts are required for modification of sensing probe which are also described. One of the applications is application to the SELEX (Systematic evolution of ligands by exponential enrichment) which is method for new aptamer generation. The principles of SELEX as well as general and modified aptamer selection method are described. In chapter 2, multi-functional biosensor was developed for chemokine ligand screening and detection in colorectal cancer cell line, where chemokine has important role in angiogenesis towards the cancer cell. Especially, the interaction between chemokine ligands and its receptor is involved in cancer metastasis. In this respect, electrochemical biosensor for chemokine screening and detection was designed in a single experimental setting. The sensing probe was prepared by anchoring the CXCR2 on the gold nanoparticles (AuNPs) deposited conducting polymer-composite layer. The interaction between CXCR2 and chemokines was studied using electrochemical impedance spectroscopy (EIS) and voltammetry. Among the chemokine ligands, CXCL5 showed the highest affinity to CXCR2, which was further used for development of an amperometric CXCL5 biosensor. Experimental parameters, in terms of concentration of receptor, pH, temperature, and incubation time were optimized. The dynamic range of CXCL5 sensor was obtained between 0.1 and 10 ng/mL with the detection limit of 0.078 ng/mL (RSD<4.7%). The proposed biosensor was successfully applied to detect CXCL5 in clinically relevant concentrations in human serum and colorectal cancer cells samples with high sensitivity and selectivity. Interference effect and the stability of the developed biosensor were also evaluated. Method verification was performed by comparing the results using commercially available ELISA kit for CXCL5 detection. In chapter 3, the magnetic force assisted electrochemical sensor (MESA) was developed using the aptamer-antibody sandwich formation to detect the thrombin as a target model protein in serum samples. The MESA detection system was used for the reaction between bioconjugates and thrombin and the removal of unbound bioconjugates from the working electrode without washing step, which detection is obtained by controlling the magnetic field. Thrombin was determined by the reduction of a toluidine blue O (TBO) and thrombin antibodies attached magnetic nanoparticle (MNP) at the sensing probe. We applied a thrombin-specific aptamer as the capture molecule which bound to the COOH-functionalized conducting polymer (poly-(2,2´:5´,5″-terthiophene-3'(p-benzoic acid) (pTBA)) layer. The streptavidin and starch coated-MNP was conjugated with biotinylated thrombin antibodies (Ab) and TBO as the bioconjugate (MNP@Ab-TBO) to detect thrombin in a serum sample. The MNP@Ab-TBO and sensing probe were characterized using voltammetry, impedance spectroscopy, XPS, and UV–VIS spectroscopy. The experimental conditions were optimized in terms of pH, binding time, removal time of unbound bioconjugates, and applied potential. The dynamic ranges of thrombin were obtained from 1.0 to 500 nM with detection limit of 0.49 nM. The recovery test demonstrates the reliability of the proposed sensing system for a handheld device. In chapter 4, the rocuronium biosensor was developed using a lipid-bonded conducting polymer with porous carbon composite in whole blood samples. Rocuronium is a neuromuscular blocking agent used with anesthetic medicine to facilitate tracheal intubation by muscle relaxation. A fast and sensitive amperometric sensor was developed for the detection of rocuronium in human whole blood samples. The sensing probe was fabricated by using phosphatidylinositol lipid (PI) bonded 3’-(2-aminopyrimidyl)-2,2’: 5’,2’’-terthiophene and porous carbon, which revealed the enhanced response to the rocuronium oxidation. The characterization of modified sensor layers was confirmed by electrochemical and surface analysis experiments. The experimental parameters affecting the rocuronium analysis were studied such as lipid and porous carbon concentration, pH, temperature, and interference species. A dynamic range was observed between 0.025 to 10 mg/mL with the detection limit of 3.83 ng/mL. The reliability of proposed sensor was successfully demonstrated via rocuronium detection in clinical samples without pre-treatment. In chapter 5, simple and fast SELEX (systematic evolution of ligands by exponential enrichment) process was developed based on an AC potential modulated electrochemical channel which modified with a functionalized-conducting polymer. The biomarker for malaria, a model target protein, was covalently bonded on the polymer coated on channel walls. The AC potential applied to carbon channel walls for inducing the specific binding between DNA molecules and target proteins by electro-kinetic mixing, and aptamers separation. In this case, the channel device achieved a partitioning efficiency of 1.67 x 107 and background of 5.56 × 10-6 through the quantitative PCR. The dissociation constants (Kd) of selected aptamers were determined employing electrochemical impedance spectroscopy (EIS) and a fluorescence method after one round selection. The PvLDH detection was carried out by aptamer array sensor assembled with selected aptamers, where signal amplification was ensured by covalent bonding of aptamer to gold nanoparticle-conducting polymer composite. This approach allowed to reach the detection limit (7.8 fM) was ~105 times lower than Kd (31.4 nM).