• Biomolecules & Therapeutics
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• 제29권2호
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• pp.144-153
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• 2021

Astrocytes play various important roles such as maintaining brain homeostasis, supporting neurons, and secreting inflammatory mediators to protect the brain cells. In aged subjects, astrocytes show diversely changed phenotypes and dysfunctions. But, the study of aged astrocytes or astrocytes from aged subjects is not yet sufficient to provide a comprehensive understanding of their important processes in the regulation of brain function. In this study, we induced an in vitro aged astrocyte model through late passage cultivation of rat primary cultured astrocytes. Astrocytes were cultured until passage 7 (P7) as late passage astrocytes and compared with passage 1 (P1) astrocytes as early passage astrocytes to confirm the differences in phenotypes and the effects of serial passage. In this study, we confirmed the morphological, molecular, and functional changes of late passage astrocytes showing aging phenotypes through SA-β-gal staining and measurement of nuclear size. We also observed a reduced expression of inflammatory mediators including IL-1β, IL-6, TNFα, iNOS, and COX2, as well as dysregulation of wound-healing, phagocytosis, and mitochondrial functions such as mitochondrial membrane potential and mitochondrial oxygen consumption rate. Culture-conditioned media obtained from P1 astrocytes promoted neurite outgrowth in immature primary cultures of rat cortices, which is significantly reduced when we treated the immature neurons with the culture media obtained from P7 astrocytes. These results suggest that late passage astrocytes show senescent astrocyte phenotypes with functional defects, which makes it a suitable model for the study of the role of astrocyte senescence on the modulation of normal and pathological brain aging.

• Parasites, Hosts and Diseases
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• 제44권1호
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• pp.15-20
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• 2006

Free-living amoebae of the genus Acanthamoeba are causative agents of granulomatous amebic encephalitis and amebic keratitis. Because the virulence of Acanthamoeba culbertsoni cultured in the laboratory is restored by consecutive brain passages, we examined the genes induced in mouse brain-passaged A. culbertsoni by differential display reverse transcriptase polymerase chain reaction (DDRT-PCR). Enhanced A. culbertsoni virulence was observed during the second mouse brain passage, i.e., infected mouse mortality increased from $5\%\;to\;70\%.$ Ten cDNAs induced during mouse brain passage were identified by DDRT-PCR and this was confirmed by northern blot analysis. BlastX searches of these cDNAs indicated the upregulations of genes encoding predictive NADH-dehydrogenase, proteasomal ATPase, and GDP-mannose pyrophosphorylase B, which have previously been reported to be associated with A. culbertsoni virulence factors.

• 대한바이러스학회지
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• 제26권1호
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• pp.67-76
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• 1996

The methods that make Hantavirus grow consist of inoculation into the experimental animals and cultured cells. The cultured cells, such as Vero-E6 and A549 cells, have been usually used for isolation of the virus and the animals, such as mice and rats, are used for large scale preparation of the virus so far. Furthermore, the cell can be used to maintain the virus and assay the infectivity and the animals can be used for the experiment of viral pathogenicity and challenge for assessment of vaccine. Apodemus mice, the own natural host of the virus, has been used for challenge test of Hantaan virus. However it has been pointed out to difficult handling and breeding the animal in laboratory. Therefore, we attempted to establish a new animal model for challenge test at the time of isolation of Maaji virus which is a new hantavirus similar but distinct to Hantaan virus. In suckling hamster, the titer of Maaji virus and the lethality to mice of the virus were increased gradually in the titer and lethality through passage by intracerebral (IC) inoculation. We tried to re-adapt this brain virus to lung of weanling hamster. The brain passaged virus was inoculated into weanling hamster intramuscularly. Again, the titer of the virus in lung was also increased by continuous passage of this virus. This facts could regarded as adaptation to new environment in which the virus proliferates. To identity the virus passaged in hamster with Maaji virus, both of the virus passaged in hamster brain and lung were compared with Maaji virus (MAA-I) and Hantaan virus (HTN 76-118) by means of restriction fragment length polymorphism (RFLP) and slingle strand conformation polymophism (SSCP). As a result, we conclude that Maaji virus could be adapted successfully to weanling hamster through this passage strategy. Utilizing this adapted Maaji virus strain, hamster model is able to be used for challenge test in hantaviral vaccinology and further experiments utilizing hamster system as a rather available and convenient lab animal are expected.

• BMB Reports
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• 제55권5호
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• pp.213-219
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• 2022

The blood-brain barrier (BBB) is an interface between cerebral blood and the brain parenchyma. As a gate keeper, BBB regulates passage of nutrients and exogeneous compounds. Owing to this highly selective barrier, many drugs targeting brain diseases are not likely to pass through the BBB. Thus, a large amount of time and cost have been paid for the development of BBB targeted therapeutics. However, many drugs validated in in vitro models and animal models have failed in clinical trials primarily due to the lack of an appropriate BBB model. Human BBB has a unique cellular architecture. Different physiologies between human and animal BBB hinder the prediction of drug responses. Therefore, a more physiologically relevant alternative BBB model needs to be developed. In this review, we summarize major features of human BBB and current BBB models and describe organ-on-chip models for BBB modeling and their applications in neurological complications.

• Parasites, Hosts and Diseases
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• 제21권2호
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• pp.234-240
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• 1983

장기간 시험관내에서 무균적으로 배양하여 병원성이 약화된 자유생활아메바 naegleria fowleri를 마우스에 연속적으로 감염시켜 그 병원성의 증감여부를 관찰하고자 하였다. 체증 18∼22gm의 백색 마우스를 secobarbital로 마취시키고 오른쪽 비강에 Naegleria fowleri를 떨어뜨려 감염시켰다. 시험관내에서 7년이상 CGVS배지에서 계대배양된 0주와 마우스에 감염시켜 뇌조직을 2번 통과시킨 2-1주의 병원성을 비교하였다. 행동둔화, 자극에 민감, 회전운동, 사지마비등 여ㅓ 증상이 나타났고, 뇌의 오른쪽 앞쪽 부위에 심한 염증 및 괴사를 발견하였다. 이러한 증상과 병소는 0주 감염군보다 2-1감염군에서 보다 빨리, 보다 심하게 발생되었음을 관찰하였다. 감염 7일 후부터 감염된 마우스가 사망하기 시작하였으며 2-1주 감염군에서 0주 감염군 보다 생존기간이 짧았으며 감염 13일 후부터 사망한 예에 있어서는 육안적으로 뇌에서 병소가 없었고 폐장에서 염증이 심하였음을 관찰하였다. 마우스에 비강을 Naegleria fowleri를 감염시켜 전형적인 원발성 아메바성 뇌수막염을 발생시킬 수 있었다. 장기간 시험관에서 무균적으로 계대배양된 Naegleria fowleri는 병원성이 약화되어 있었고 이 아메바를 마우스에 연속적으로 감염시켜 뇌조직을 통과시킴으로 다시 병원성이 증강됨을 관찰하였다.

• 대한미생물학회지
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• 제16권1호
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• pp.83-89
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• 1981

Japanese encephalitis has been prevalent for long time in the Far East and many patients have been reported in both South East and Mid-West Asia recently. Recently, vaccine was used in prevention of this viral disease of man which was derived from formalin inactivated virus inoculated into mouse brain, but live attenuated active vaccine for human is not developed yet. Author inoculated Japanese encephalitis virus into several cell culture strains for development of live attenuated encephalitis virus strain and the results were as follows: 1. Japanese encephalitis virus was inactivated rapidly in cell free medium at $36^{\circ}C$ and totally inactivated by 72 hours. 2. In growth curve of Japanese encephalitis virus in HeLa cell cultures, maximal multiplication of the virus was occured at 4th day and virus multiplication was continued for at least 12 days. 3. After succeeding passage of the virus in HeLa cell cultures and human esophagus epithelial cell cultures, infectivity of virus for mice was disappeared from 2nd passage in HeLa cell cultures and 3rd passage in esophagus epithelial cell cultures. 4. In inoculation to monkey kidney epithelial cells and chick embryo cell cultures, infectivity of the virus for mice was continued after 10th passages.

• 핵의학기술
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• 제23권1호
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• pp.69-74
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• 2019

$^{18}F$-FDOPA는 뇌 종양의 아미노산 대사를 추적하는 방사성 의약품이다. 본 연구의 목적은 뇌 종양의 아미노산 대사를 영상화 하여 악성 종양을 진단하는 $^{18}F$-FDOPA와 포도당 대사를 통한 $^{18}F$-FDG의 Brain PET/CT 검사 영상의 대조도 분석을 통해 병변의 검출 능력을 비교하고, $^{18}F$-FDOPA Brain PET/CT 검사에서 섭취 시간에 따른 SUV의 변화를 분석하여 최적의 영상 획득 시간을 알아보기 위함이다. $^{18}F$-FDOPA 와 $^{18}F$-FDG 두 영상에서 종양(Tumor)과 소뇌(Cerebellum)의 중심에 각각 약 $350mm^2$의 관심 영역을 설정하여 $SUV_{max}$를 측정하였고, 종양과 소뇌의 $SUV_{max}$ 비율(T/C ratio)을 산출하였고, $^{18}F$-FDOPA 투여 직후 30분 동안 획득한 리스트 수집 방식 데이터(List mode data)를 활용해 2분씩 15프레임으로 나눈 뒤 각 프레임 별로 종양과 소뇌 중심에 $SUV_{max}$를 측정하여 위와 동일한 방법으로 T/C ratio를 산출하여 분석하였다. 종양의 평균 $SUV_{max}$를 비교해 본 결과, $^{18}F$-FDOPA Brain PET/CT 검사에서 $4.2{\pm}0.8$, $^{18}F$-FDG Brain PET/CT 검사에서는 $5.6{\pm}0.7$ 이었다. 또한, T/C ratio는 $^{18}F$-FDOPA 검사에서 $2.1{\pm}0.7$, $^{18}F$-FDG 검사에서는 $1.1{\pm}0.4$ 이었으며, $^{18}F$-FDOPA의 $SUV_{max}$는 $^{18}F$-FDG보다 낮지만 T/C ratio는 높게 나타나 종양 구별 능력이 더욱 뛰어난 것을 알 수 있었다(t=-5.214, p=0.000). $^{18}F$-FDOPA의 섭취 시간에 따른 $SUV_{max}$와 T/C ratio를 분석한 결과, $SUV_{max}$와 T/C ratio의 Peak는 모두 6~8분에서 나타났다. 이를 토대로 본원에서 $^{18}F$-FDOPA Brain PET/CT 검사에서 활용하는 10~30분의 영상과 Peak가 나타나기 시작한 6~26분의 영상을 비교한 결과 SUV와 T/C ratio가 각각 0.2, 0.1 증가하였다. 추후 지속적인 연구를 통해 검사 소요시간의 단축 가능성과 추가적인 스캔 정보 활용을 통한 정확한 진단에도 도움이 될 것으로 사료된다.

• Parasites, Hosts and Diseases
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• 제47권2호
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• pp.103-107
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• 2009

The genus Acanthamoeba can cause severe infections such as granulomatous amebic encephalitis and amebic keratitis in humans. However, little genomic information of Acanthamoeba has been reported. Here, we constructed Acanthamoeba expressed sequence tags (EST) database (Acanthamoeba EST DB) derived from our 4 kinds of Acanthamoeba cDNA library. The Acanthamoeba EST DB contains 3,897 EST generated from amebae under various conditions of long term in vitro culture, mouse brain passage, or encystation, and downloaded data of Acanthamoeba from National Center for Biotechnology Information (NCBI) and Taxonomically Broad EST Database (TBestDB). The almost reported eDNA/genomic sequences of Acanthamoeba provide stand alone BLAST system with nucleotide (BLAST NT) and amino acid (BLAST AA) sequence database. In BLAST results, each gene links for the significant information including sequence data, gene orthology annotations, relevant references, and a BlastX result. This is the first attempt for construction of Acanthamoeba database with genes expressed in diverse conditions. These data were integrated into a database (http://www. amoeba.or.kr).

• 한국디지털건축인테리어학회논문집
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• 제12권4호
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• pp.39-48
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• 2012

Generally, rural housing in Korea have had target of people who are in good physical health rather than the disabled. Therefore, it is difficult to offer high quality residential environment for the elderly and the disabled. The purpose of this study is to present the remodeling of rural house for the disabled. this study divides the a rural house into 7 sectors : a passage to entrance, an entrance hall, a livingroom, a bedroom, a bathroom, a kitchen, and etc. this study propose the following renovation plan that rural housing to be barrier free space. First, in passage to entrance of the house, people with disabilities should not experience difficulties in walking, so installing ramps to remove the stepped slope grade without slip so that the floor finish should be. Second, install grab handle on the wall inside the housing so that the disabled can lean on the handle. Third, placing the furniture in the bedroom that wheelchair can be rotated, and make to lower the height of the switchs. Forth, install a non-slip floor tile to prevent accidents in the bathroom.

• 한국발생생물학회:학술대회논문집
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• 한국발생생물학회 2003년도 제3회 국제심포지움 및 학술대회
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• pp.105-105
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• 2003

In previous reports, pVPSV.IGR2.1 transgenic mouse were described that brain tumor and lymphoma by reason of Vasopressin-SV40 T antigen. In this study, we produced pVPSV.IGR3.6 transgenic mouse that used pVPSV.IGR3.6 vector. Expression of transgene was vary different in transgenic mouse. We obtained 6 transgenic mouse line, moreover they had died at the age of 2-6 weeks without transmitting the transgene to their offspring, and had tumorigenesis on same location with pVPSV.IGR2.1 transgenic mouse. Only a founder mouse was investigated for expression of fusion gene. Here we extended this transgenic approach to the study of tumor progression. From the mouse, we confirmed brain tumor cell, after then cultured for investigate characterization. In this report, we demonstrate that reduction of survival rate in transgenic mouse fused vasopressin gene length, acquisition of brain tumor cell, composition with astrocyte cells and neuronal cells. Finally, cells had no change with increase of passage.