• Biomolecules & Therapeutics
• /
• v.29 no.2
• /
• pp.144-153
• /
• 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
• /
• v.44 no.1 s.137
• /
• pp.15-20
• /
• 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.

• The Journal of Korean Society of Virology
• /
• v.26 no.1
• /
• pp.67-76
• /
• 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
• /
• v.55 no.5
• /
• pp.213-219
• /
• 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
• /
• v.21 no.2
• /
• pp.234-240
• /
• 1983

The pathogenicity of free-living amoeba, Waegleria fcwleri, is influenced according to the strain, cultural condition and host (Culbertson et at., 1968; Carter, 1970; Wong et at., 1975), Phillips (1973) demonstrated that Entamoeba histolytica became avirulent after more than 2 year maintenance in axonic culture in vitro. This study was carried out to compare the difference in pathogenicity between two strains of N. fowleri, one of a prolonged maintenance in arsenic medium and the other one obtained by serial brain passage in mice. The 0 strain was that N. fowleri had cultivated axenically more than 7 years in CGVS medium. The 2-1 strain was obtained from the brain of mouse inoculated intranasally with a strain, which was from the mouse brain infected with 0 strain, and cultured for 15 weeks until the beginning of this experiment. White male mice weighing 18-22 g were used. Mice were anesthetized by an intraperitoneal injection of about 1 mg secobarbital, and inoculated intranasally with $10{\times}$10^4 live N. fowleri trophoBoites in a $5{\;}{\mu}l$ cell suspension. Sluggish behaviour, nervousness, rotation and leg paralysis were developed earlier and more frequently in the 2-1 experimental group than the control 0 group. Pathological changes such as inflammatory and necrotic lesion were observed in the olfactory and anterior portion of brain, and these changes were more extensive in the 2-1 group. The edematous and inflammatory changes in lung were demonstrated in mice died after 13th day post-inoculation. The experimental mice of 2-1 group began to die suddenly from 7th day post-inoculation, and the survival time in 2-1 group mice was shorter than 0 group mice. The typical primary amoebic meningoencephalitis was developed in the mice inoculated intranasally with N. fowleri. The prolonged maintenance of N. fowleri amoebae in axonic CGVS medium was observed to have lost their original pathogenicity for mice, but their pathogenicity was restored by serial brain passage in mice.

• The Journal of the Korean Society for Microbiology
• /
• v.16 no.1
• /
• pp.83-89
• /
• 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.

• The Korean Journal of Nuclear Medicine Technology
• /
• v.23 no.1
• /
• pp.69-74
• /
• 2019

Purpose $^{18}F$-FDOPA using amino acid is particularly attractive for imaging of brain tumors because of the high uptake in tumor tissue and the low uptake in normal brain tissue. But, on the other hand, $^{18}F$-FDG is highly uptake in both tumor tissue and normal brain tissue. The purpose of study is to evaluate comparison of contrasts in $^{18}F$-FDOPA Brain PET/CT and $^{18}F$-FDG Brain PET/CT and to find out optimal scan time by analysis of variation in SUV with the passage of uptake time. Materials and Methods A region of interest of approximately $350mm^2$ at the center of the tumor and cerebellum in 12 patients ($51.4{\pm}12.8yrs$) who $^{18}F$-FDG Brain PET/CT and $^{18}F$-FDOPA Brain PET/CT were examined more than once each. The $SUV_{max}$ was measured, and the $SUV_{max}$ ratio (T/C ratio) of the tumor cerebellum was calculated. In the analysis of SUV, T/C ratio was calculated for each frame after dividing into 15 frames of 2 minutes each using List mode data in 25 patients ($49.{\pm}10.3yrs$). SPSS 21 was used to compare T/C ratio of $^{18}F$-FDOPA and T/C ratio of $^{18}F$-FDG. Results The T/C ratio of $^{18}F$-FDOPA Brain PET/CT was higher than the T/C ratio of $^{18}F$-FDG Brain, and show a significant difference according to a paired t-test(t=-5.214, p=0.000). As a result of analyzing changes in $SUV_{max}$ and T/C ratio, the peak point of $SUV_{max}$ was $5.6{\pm}2.9$ and appeared in the fourth frame (6 to 8 minutes), and the peak of T/C ratio also appeared in the fourth frame (6 to 8 minutes). Taking this into consideration and comparing the existing 10 to 30 minutes image and 6 to 26 minutes image, the $SUV_{max}$ and T/C ratio increased by 0.2 and 0.1 each, compared to the 10 to 30 minutes image for 6 to 26 minutes image. Conclusion From this study, $^{18}F$-FDOPA Brain PET/CT is effective when reading the image, because the T/C ratio of $^{18}F$-FDOPA Brain PET/CT was higher than T/C ratio of $^{18}F$-FDG Brain PET/CT. In addition, in the case of $^{18}F$-FDOPA Brain PET/CT, there was no difference between the existing 10 to 30 minutes image and 6 to 26 minutes image. Through continuous research, we can find possibility of shortening examination time in $^{18}F$-FDOPA Brain PET/CT. Also, we can help physician to accurate reading using additional scan data.

• Parasites, Hosts and Diseases
• /
• v.47 no.2
• /
• pp.103-107
• /
• 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).

• Journal of The Korean Digital Architecture Interior Association
• /
• v.12 no.4
• /
• pp.39-48
• /
• 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.

• Proceedings of the Korean Society of Developmental Biology Conference
• /
• 2003.10a
• /
• pp.105-105
• /
• 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.