• Title/Summary/Keyword: Simple Technique

Search Result 3,403, Processing Time 0.028 seconds

Neuroglial Reaction in the Substantia Nigra and Striatum of 6-Hydroxydopamine Induced Parkinson's Disease Rat Model (흰쥐 흑질내 수산화도파민 주입으로 유도된 파킨슨병 모델에서 흑질과 선조체의 신경교세포 반응)

  • Yang, Kyung Won;Sung, Jae Hoon;Kim, Moon Chan;Lee, Moon Yong;Lee, Sang Won;Choi, Seung Jin;Park, Choon Keun;Kang, Joon Ki
    • Journal of Korean Neurosurgical Society
    • /
    • v.30 no.6
    • /
    • pp.688-698
    • /
    • 2001
  • Objectives : Parkinson's disease is a well-known neurodegenerative disease characterized by dopaminergic cell death in the substantia nigra. The reactive gliosis by activated astrocytes and microglias is no more regarded as a simple sequel of neuronal cell death. Microglial activation takes place in a stereotypic pattern with graded morphologic and functional(resting, activated and phagocytic) changes. In Parkinson's disease animal model, the degree of microglial activation along the nigro-striatal dopaminergic tract has not been studied intensively. The purpose of this study was to elucidate the characteristics of microglial reaction and to grade its degree of activation at substantia nigra and corpus striatum using 6-hydroxydopamine induced rat model of Parkinson's disease. Methods : Using Sprague-Dawley rat, parkinsonian model was made by 6-hydroxydopamine(OHDA) induced destruction of medial and lateral substantia nigra(SN). The rat was sacrificed 3-, 5-, 7-, 14- and 21-day-after operation. For control group, we injected saline with same manner and sacrificed 3-day after operation. With immunohistochemistry, we examined dopaminergic neuronal cells and microglial expression using tyrosine hydroxylase (TH) and OX-42 antibodies, respectively. Also we performed in situ hybridization for osteopontin, a possible marker of subset in activated microglia. Results : 1) In lesioned side of substantia nigra and corpus striatum, the TH immunoreactivity was markedly decreased in whole experimental groups. 2) Using optical densitometry, microglia induced immunoreactivity of OX-42 was counted at SN and corpus striatum. At SN, it was increased significantly on the lesioned side in control and all time-dependent experimental groups. At striatum, it was increased significantly in post lesion 3-day group only(p <0.05). Compared to control group, immunoreactivity of OX-42 on lesioned side was increased in groups, except post lesion 21-day group, at SN. Only post lesion 3-day group showed significance at striatum(p <0.05). Compared to SN region, immunoreactivity of OX-42 was much weaker in striatum. 3) Microscopically, the microglias showed typically different activation pattern. At SN, numerous phagocytic microglias were found at pars compacta and reticularis of lesion side. At striatum, no phagocytic form was found and the intensity of staining was much weaker. 4) At SN, the immunoreactivity of osteopontin showed definite laterality and it was markedly increased at pars compacta of lesion side with relatively short duration time. At striatum, however, it was not detected by in situ hybridization technique. Conclusion : The nigral 6-OHDA induced rat model of Parkinson's disease revealed several characteristic patterns of microglial reaction. At SN, microglias was activated shortly after direct neuronal damage and maintained for about three weeks. In contrast, despite of sufficient dopaminergic insufficiency at striatum, activation of microglias was trivial, and distinguished 3 day later. Antegrade slow neuronal degeneration is major pathophysiology in striatal dopaminergic deficiency. So, the acuteness of neuronal damage and consequential degree of neuronal degeneration may be important factor for microglial activation in neurodegenerative diseases such as Parkinson's disease. Additionally, osteopontin may be a possible marker for several subsets of activated microglia, possibly the phagocytic form.

  • PDF

The Effects of Polyampholyte on Vitrification Process for cryopreservation of Bovine Oviduct Epithelial Cell (Polyampholyte가 소난관상피세포의 초자화 동결방법에 미치는 영향)

  • Kim, Sung Woo;Lee, Jae-Yeong;Kim, Chan-Lan;Yu, Yeonhee;Lee, Sung Soo;Ko, Yeoung-Gyu
    • Journal of the Korea Academia-Industrial cooperation Society
    • /
    • v.21 no.6
    • /
    • pp.527-535
    • /
    • 2020
  • The purpose of this study was to establish a simple vitrification protocols to preserve animal cell lines derived from tissues of livestock that could be recultured. Bovine oviduct epithelial cells (BOEC) were used for the vitrification process using a 0.25 ml straw to increase cryopreservation efficiency. BOEC was cultured from the oviduct of 3.5-day estrus state, and the commercially available polyampholyte StemCell KeepTM was used as a cryoprotective agent. Using different concentrations, the viability rates of BOEC in 5, 10, 25, 50, 75, and 100% in freezing media were investigated. Survivability was determined using a differential staining technique using a trypan blue test and a CYTO-13/PI staining protocol. The viability rates of BOEC in the trypan blue test were 5.6±11.8, 12.5±7.2, 53.0±2.7, 85.1±6.9, 79.8±0.6, and 60.7±6.7% with a respective concentration of StemCell KeepTM. The viability rates in CYTO-13/PI staining were 4.6±2.5, 30.8±12.1, 58.4±2.5, 85.5±1.2, 79.8±0.6, and 71.2±1.2%, respectively. These results indicate that BOEC could be preserved with StemCell KeepTM without toxicity in a 0.25-ml straw. The optimal concentration of vitrification solution with StemCell KeepTM was determined to be 50% and can be considered as a proper preservation method for cryobanking.

Arthroscopic Treatment of Metallic Suture Anchor Failures after Bankart Repair (Bankart 수술 후 발생한 금속 봉합 나사못 합병증의 관절경적 치료)

  • Shin, Sang-Jin;Jung, Jae-Hoon;Kim, Sung-Jae;Yoo, Jae-Doo
    • Journal of the Korean Arthroscopy Society
    • /
    • v.10 no.1
    • /
    • pp.70-76
    • /
    • 2006
  • Purpose: This study presents 5 patients who had metallic anchor protrusion on glenoid after Bankart repair in anterior shoulder instability and reviewed the cause, clinical feature and arthroscopic removal technique. Method and Materials: 5 male with average age of 22 years (range 19 to 25 years) were included. 4 patients had arthroscopic Bankart repair and 1 patient had open repair for anterior shoulder instability. They had protruded metallic suture anchors on glenoid and the protruded suture anchors were removed arthroscopically using larger suture anchor empty inserter. Results: 4 patients had painful clicking sound with motion of abduction and external rotation and 1 patient showed shoulder instability. The ROM showed normal except mild degrees loss of external rotation. The position of protruded metallic anchor was 2, 3 and 5 O'clock in three patients and 4 O'clock in 2 patients. In 2 patients, the metallic suture anchor was malpositioned about 5mm off on the medial side from the anterior glenoid edge. All had Outerbrige classification Grade II-III chondral damage on humeral head and 1 patient showed glenoid cartilage destruction. None had shoulder instability after 2 years of follow-up. Constant score was 65 preoperatively and 89 postoperatively. ASES score was 67 preoperatively and 88 postoperatively. Conclusion: Symptoms of protruded suture anchor are not combined with instability. Most of symptoms were revealed from the rehabilitation period and confused with postoperative pain. Prompt diagnosis and early arthroscopic removal or impaction of protruded metallic suture anchor is recommended because of serious glenohumeral cartilage destruction. This is easy and simple and reproducible method to remove protruded metallic suture anchor arthroscopically.

  • PDF

ELECTRON MICROSCOPIC STUDY OF SLOWLY ADAPTING PERIODONTAL MECHANORECEPTIVE PRIMARY AFFERENT FIBERS WITHIN THE SUBNUCLEUS ORALIS OF THE CAT (서순응형 치근막 일차구심성 신경섬유 종말부의 Subnucleus oralis에서의 시냅스 양상에 관한 전자현미경적 연구)

  • Kim, Moo-Jung;Bae, Yong-Chul;Kim, Chin-Soo
    • Maxillofacial Plastic and Reconstructive Surgery
    • /
    • v.15 no.4
    • /
    • pp.281-301
    • /
    • 1993
  • It was revealed that the morphology and projection pattern of terminal arbors from single primary afferent are different among distinct fiber types, functional types and the different subdivision of trigeminal sensory nucleus complex(TSNC). But it was not identified the ultrastructural morphology and synaptic connections of terminal arbors from each primary afferent within TSNC. So we employed the intra-axonal horseradish peroxidase(HRP) injection technique to define the terminal arbors of primary afferent fiber from slowly adapting mechanoreceptors in the periodontal ligament of the cat, and examined 66 labeled terminal arbors within the rostrodorsomedial part(Vo.r) of the trigeminal nucleus oralis, electromicroscopically with 90nm serial sections. All the boutons labelled with HRP contained clear, spherical and uniform sized synaptic vesicles(diameter : $47.66{\pm}3.58nm$ ). Most of the labelled boutons were boutons en passant type and they were connected by unmyelinated axonal strand. In which neurofilament and microtubule was not developed but occasionally contained synaptic vesicle in contrast to the myelinated axon. The size of the labelled bouton was relatively small(long diameter : $1.46{\pm}0.24{\mu}m$, short diameter $0.85{\pm}0.26{\mu}m$, average diameter $1.15{\pm}0.24{\mu}m$) and the shape of which varied from dome to elongated shape, but scalloped glomerulus shape was not developed. Each primary ending in Vo.r made synapse with one or two neuronal propiles(average : $1.11{\pm}0.31$), of which, 89.4% of labelled boutons made synapse with only one neuronal pro pile, the remainder, 10.6% of labelled boutons, made synapse with two neuronal propile. So characteristically they made very simple synapse. Most of labelled boutons(80.03%) made asymmetrical synapse only with dendritic shaft or spine, and 6.1% of labelled boutons received symmetrical synapse from pleomorphic vesicle containing axonal ending(p-ending). So presynaptic inhibiton was relatively scarce. Synaptic triad, in which a p-ending is presynaptic both pre-and post-synaptic element of the axo-dendritic contact from the labelled primary ending was not observed.

  • PDF

Speed-up Techniques for High-Resolution Grid Data Processing in the Early Warning System for Agrometeorological Disaster (농업기상재해 조기경보시스템에서의 고해상도 격자형 자료의 처리 속도 향상 기법)

  • Park, J.H.;Shin, Y.S.;Kim, S.K.;Kang, W.S.;Han, Y.K.;Kim, J.H.;Kim, D.J.;Kim, S.O.;Shim, K.M.;Park, E.W.
    • Korean Journal of Agricultural and Forest Meteorology
    • /
    • v.19 no.3
    • /
    • pp.153-163
    • /
    • 2017
  • The objective of this study is to enhance the model's speed of estimating weather variables (e.g., minimum/maximum temperature, sunshine hour, PRISM (Parameter-elevation Regression on Independent Slopes Model) based precipitation), which are applied to the Agrometeorological Early Warning System (http://www.agmet.kr). The current process of weather estimation is operated on high-performance multi-core CPUs that have 8 physical cores and 16 logical threads. Nonetheless, the server is not even dedicated to the handling of a single county, indicating that very high overhead is involved in calculating the 10 counties of the Seomjin River Basin. In order to reduce such overhead, several cache and parallelization techniques were used to measure the performance and to check the applicability. Results are as follows: (1) for simple calculations such as Growing Degree Days accumulation, the time required for Input and Output (I/O) is significantly greater than that for calculation, suggesting the need of a technique which reduces disk I/O bottlenecks; (2) when there are many I/O, it is advantageous to distribute them on several servers. However, each server must have a cache for input data so that it does not compete for the same resource; and (3) GPU-based parallel processing method is most suitable for models such as PRISM with large computation loads.

Enhancement of the solubility of human tissue inhibitor of matrix metallocroteinase-2 (TIMP-2) in E. coli using a modified in vitro mutagenesis (새로운 유전자 재조합 방법을 이용한 대장균에서의 인간 tissue inhibitor of mtrix metalloproteinase-2 (TIMP-2) 유전자의 가용성 발현)

  • Kim, Jong-Uk;Choi, Dong-Soon;Joo, Hyun;Min, Churl-K.
    • KSBB Journal
    • /
    • v.23 no.3
    • /
    • pp.231-238
    • /
    • 2008
  • The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

A Study on eDesign Platform for Effective Communication and Information sharing - with an emphasis on process and template (효과적인 커뮤니케이션과 정보공유를 위한 e디자인 플랫폼 구축에 관한 연구 - 프로세스와 템플릿을 중심으로)

  • 윤주현
    • Archives of design research
    • /
    • v.17 no.2
    • /
    • pp.425-436
    • /
    • 2004
  • A new design field called eDesign appears as if eBiz is an online related business in which an industrialized design creates the individual value added facing a digital revolution. The field of eDesign requires a special design process and management methodology regardless of the fact that human sensitivity will be satisfied through a dehumanized computer technique. However, it is the reality of eDesign that has been dependent upon a simple process or project management tool of general design. In this study, we develop an eDesign platform based on an eDesign process and template mainly focused on eBusiness in order to overcome the wrong situation. The template is a kind of document that has a standardization form. We aim to establish a general process through various case projects, store information using a necessary template, and use for the way of visual communication. We propose a standard of eDesign platform that can be widely applied to the field of design, medium and small enterprises focused on IT businesses or design-team through this project performed as an educational-industrial study. It makes it possible to get a detailed process methodology, which can be applied to many small design related companies that don't have their own process yet, and will be a scale for comparing their own process in which the company has a process of opened standard eDesign with it. In addition, it makes possible a systematic control of the own projects within and outside the firm, accumulating information for the firm through the database, and easy communication. Furthermore, it can be applied to check the process of the project as a checklist, and then it will reduce trial and error repeated for every project that has been done.

  • PDF

A Study of Genetic Polymonhisms of HLA-class I and II Genes Using Polymerase Chain Reaction (중합효소연쇄반응을 이용한 HLA-class I, II 유전자군의 유전적 다형성에 관한 연구)

  • Kyung-Ok Lee
    • Biomedical Science Letters
    • /
    • v.4 no.1
    • /
    • pp.11-25
    • /
    • 1998
  • The HLA genes located in the short arm of chromosome 6 specify heterodimeric glycoproteins involved in the regulation of the immune response. Recently, in the elucidation of HLA polymorphism, serological and cellular typing methods have been replaced by DNA typing using polymerase chain reaction (PCR). The purpose of this study was to establish the HLA DNA typing methods and determine gene frequencies of HLA molecules in Koreans. PCR-SSP (sequence specific primers) and PCR-RFLP (restriction fragment length polymorphism) techniques were used for the analysis of HLA-A, -B, -C, DRBl genes and HLA-DQAl, DQBl, DPBl genes, respectively. The results of B-lymphoblastoid cells used for control experiment were consistent with the previous data identified in the 11th International Histocompatibility Workshop. Seventeen, 23, 16, 8, 16, 13 and 37 types of HLA-A, B, C, DQAl, DQBl, DPBl and DRBl alleles were found, respectively, in a total of unrelated 120 Korean individuals. The most frequent HLA alleles were $A^*$02 (27.0%), B$^*$40 (17.6%), Cw$^*$01 (19.2%), DQAl$^*$0301 (32.1%), DQBl$^*$0303 (12.9%), DPBl$^*$0501 (31.3%) and DRBl$^*$1501 (9.2%) among Koreans. This study shows that DNA typing method using PCR technique is a relatively simple, fast and practical tool for the determination of the HLA-class I and II genes. Moreover, the data of HLA gene frequencies could be useful for the Korean database before clinical applications, including organ and unrelated bone marrow transplantation, anthropological study, disease association and individual identification.

  • PDF

Development of an Efficient Screening System for Resistance of Watermelon Plants to Didymella bryoniae (수박 덩굴마름병에 대한 효율적인 저항성 검정 방법 개발)

  • Lee, Ji Hyun;Jang, Kyoung Soo;Choi, Yong Ho;Kim, Jin-Cheol;Choi, Gyung Ja
    • Research in Plant Disease
    • /
    • v.22 no.2
    • /
    • pp.72-80
    • /
    • 2016
  • Gummy stem blight, caused by the fungus Didymella bryoniae, is major disease of watermelons worldwide. The objective of the present study was to establish an efficient screening system to identify watermelon resistant to D. bryoniae. An GSB3 isolate was prepared from a watermelon plant showing typical symptoms of gummy stem blight in Haman-gun and identified as D. bryoniae based on molecular analysis of internal transcribed spacer sequence. A simple mass-production technique of inoculum was developed based on spore production of D. bryoniae GSB3 under several incubation conditions and their virulence on watermelon plants. Resistance degrees of 22 commercial watermelon cultivars to the GSB3 isolate were evaluated. Among them, four watermelon cultivars showing different degree of resistance response were selected for further study. Development of disease on the cultivars according to various conditions including inoculum concentrations, incubation periods in dew chamber, and incubation temperatures was investigated. From the results, we suggest an efficient screening method for resistant watermelon cultivars to gummy stem blight. Seeds of watermelon cultivar are sown and grown in a greenhouse until plant stage of 2-fully expanded leaves. Seedlings are inoculated with D. bryoniae by spraying spore suspension of the fungus at a concentration of $5.0{\times}10^5spores/ml$. The infected plants are incubated in humidity chamber at $25^{\circ}C$ for 48 hours and then transferred to a growth chamber at $25^{\circ}C$ and 80% relative humidity with 12-hour light a day. Three to four days after inoculation, disease severity of the plant are measured using percentage of infected leaf area.

Antimycotic Susceptibility Testing of Trichophyton Rubrum by Microculture Method (96-well microplate를 이용한 Trichophyton Rubrum의 항진균제 감수성검사)

  • Lee, Moo-Woong;Kim, Jong-Chul;Choi, Jong-Soo;Kim, Ki-Hong
    • Journal of Yeungnam Medical Science
    • /
    • v.9 no.2
    • /
    • pp.396-406
    • /
    • 1992
  • Various susceptibility tests have been used to determine minimal inhibition concentration(MIC) of dermatophytes. They have limitations to apply practically because they need long time to determine MIC. Authors examined MIC of T. rubrum to ketoconazole and itraconazole using 96-well microplate and 24-well macroplate by method of Granade and Artis and tried to check the possibility of this method on clinical application. Nine strains of T. rubrum from patients with dermatophytosis were used. Evaluations of the factors affecting MIC were also tried. The results were as follows. 1. Effect of inoculation density on determination time and MIC : Determination of MIC were possible in 4th days after inoculation at higher inoculation density Caborbance 2.0, 1.0) compared to 6th days at lower inoculation density(absorbance 0.5, 0.25). 2. Effect of incubation temperature on MIC : When incubating at $37^{\circ}C$, MIC were below 0.006-$0.04{\mu}g/ml$ to ketokckonazole and below 0.006-$0.04{\mu}g/ml$ to itraconazole while at $25^{\circ}C$ 0.08-$5.68{\mu}g/ml$ to ketoconazole and 0.006-$0.71{\mu}g/ml$ to itraconazole. Significant reduction of MIC was observed at $37^{\circ}C$ compared to $25^{\circ}C$. 3. Effect of container size on determination time and MIC : When incubating in 96-well microplate and 24-well macroplate, determination of MIC was possible in 4th to 6th days after inoculation in broth-containig 96-well microplate compared to 8th to 12th days in broth-containing 24-well macroplate. But no difference in MIC was observed between different container size. 4. Effect of media on MIC : When using broth as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconazole, below 0.006-$0.36{\mu}g/ml$ to itraconazole in broth-containg 24-well macroplate. When using agar as media, MIC were below 0.006-$5.68{\mu}g/ml$ to ketoconzole, below 0.006-$5.68{\mu}g/ml$ to intraconzole in agar-containing 24-well macroplate. There was slight increase of MIC with agar media compared to broth media. 5. These findings confirm that determination of MIC of dermatophtes by method of Granade and Artis is fast and simple technique for antifungal susceptibility test.

  • PDF