• Title/Summary/Keyword: passage (in mice)

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Changes in the pathogenicity of Naegleria fowleri by serial brain passage in mice (자유생활아메바 Naegleria fowleri의 계대감염에 의한 병원성의 변화에 관한 연구)

  • 이득기;임경일
    • Parasites, Hosts and Diseases
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    • v.21 no.2
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    • pp.234-240
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    • 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.

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Passage and Adaptation of Maaji Virus in Hamster (Maaji Virus의 Hamster 계대 및 적응)

  • Kim, Yun-Cheol;Paik, Woo-Hyun;Lee, Pyung-Woo
    • The Journal of Korean Society of Virology
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    • v.26 no.1
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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.

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Propagation and Attenuation of Japanese Encephalitis Virus in Tissue Culture Cells (조직배양세포에서의 일본뇌염virus 증식에 관한 연구)

  • Lee, Ho-Wang;Moon, Seok-Bae
    • The Journal of the Korean Society for Microbiology
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    • v.16 no.1
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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.

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Tumor-Suppressing Effects of Saponin-SOD in Nude Mice (Saponin-SOD가 SNUC-4 Cell에 의해 유발편 대장암 Nude Mice의 암 성장 억제 효과)

  • 박수진;강명화
    • Journal of the East Asian Society of Dietary Life
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    • v.14 no.4
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    • pp.338-342
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    • 2004
  • This study was performed to measure the antioxidant effects of red ginseng extracts which antioxidation had been promoted through enzyme hydrolysis. In order to observe their tumor-suppressing effects, an anti-cancer medicine and Saponin-SOD, which was a highly antioxidant beverage made from red ginseng saponin adding SOD-like rice (with embryo buds) extracts, were administered to nude mice with large intestine cancer induced. There was a significant increase in the content of phenolic compounds as the enzyme was added. The red ginseng extracts showed a high electron-donating ability with the passage of time. The electron-donating ability was particularly high in the enzyme-treated red ginseng extract, and also observed as high in Saponin-SOD. The lipid-peroxide generation was inhibited depending on the concentration of Saponin-SOD added; the addition of 0.625% Saponin-SOD served to decrease the inhibition level up to 65% compared with the case of no addition (100%). As a result, it could be assumed that Saponin-SOD would strongly inhibit the oxidation of ghost membrane. After the cancer was induced in nude mice through the injection of SNUC-4 cell, there was a significant inhibition in the growth of tumors in nude mice into which Saponin-SOD were injected; the growth of tumors was gradually decreasing with the passage of time after the cancer induction. In particular, when Saponin-SOD was administered together with an anti-cancer medicine, the synergic effect was observed. In conclusion, Saponin-SOD, when used with an anti-cancer medicine, is expected to reduce the amount of free radical and lipid peroxide, which are known to cause harmful effects occurring from the internal application of medicine.

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Macrophages Promote Coal Tar Pitch Extract-induced Tumorigenesis of BEAS-2B Cells and Tumor Metastasis in Nude Mice Mediated by AP-1

  • Zhang, Peng;Jin, Yue-Fei;Zhang, Qiao;Wu, Yi-Ming;Wu, Wei-Dong;Yao, Wu;Wu, Yong-Jun;Li, Zhi-Tao;Zhao, Yong;Liu, Yu;Feng, Fei-Fei
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.12
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    • pp.4871-4876
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    • 2014
  • Background: We sought to evaluate the role of tumor associated macrophages (TAMs) on the promotion of coal tar pitch extract (CTPE)-induced tumorigenesis of human bronchial epithelial cells (BEAS-2B) and tumor metastasis in nude mice, and related mechanisms. Materials and Methods: BEAS-2B cells were first treated with 2.4 mg/mL CTPE for 72 hours. After removal of CTPE, the cells were continuously cultured and passaged using trypsin-EDTA. THP-1 cells were used as macrophage-like cells. BEAS-2B cells under different conditions (n=6/group) were injected into the back necks of nude mice, and alterations of tumor xenograft growth, indicative of tumorigenicity, and tumor metastasis were determined. Pathological changes (tumor nests and microvascular lesions) of HE-stained tumor tissues were also evaluated. The expression of AP-1(c-Jun) in xenografts and metastatic tumors was determined using immunohistochemistry. Results: Tumor size and weight in nude mice transplanted with the mixture of CTPE-induced passage 30 BEAS-2B and THP-1 cells (2:1) were increased compared to those from the CTPE-treated BEAS-2B cells at passage 30 alone at different observation time points. Tumor metastasis to lymph nodes and liver was only detected after transplantation of a mixture the two kinds of cells. The numbers of tumor nests and microvascular lesions, and the expression levels of AP-1 (c-Jun) in tumors from the mixture of two kinds of cells were increased apparently in contrast to those in tumor from the CTPE-treated BEAS-2B cells of passage 30 alone. In addition, there was positive correlation between AP-1 (c-Jun) expression level and the number of microvascular lesions, or between AP-1 (c-Jun) expression level and tumor metastasis in these two groups. Conclusions: TAMs not only facilitate tumorigenesis transformation of CTPE-induced BEAS-2B cells, but also promote tumor growth, angiogenesis and metastasis in nude mice in vivo, which may be mediated by AP-1.

Regulatory roles of NKT cells in Anaplasma phagocytophilum infection

  • Choi, Kyoung-Seong;Chae, Joon-Seok
    • Korean Journal of Veterinary Research
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    • v.49 no.2
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    • pp.167-172
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    • 2009
  • Human granulocytic anaplasmosis (HGA) is caused by the obligate intracellular bacterium Anaplasma (A.) phagocytophilum. Natural killer T (NKT) cells are key players in host defense against various microbial infections. We investigated the role of NKT cells in immune response to A. phagocytophilum infection using NKT-knockout ($J\alpha$18-/-) mice. $J\alpha$18-/- and wild-type (WT) mice were infected with low-passage A. phagocytophilum and assayed for hepatic histopathology and cytokine production during 7 days post-infection. Compared to WT controls, the infected $J\alpha$18 -/- mice had much less histopathologic lesions and less apoptosis through day 7, and lower concentrations of ${IFN\gamma}$ and IL- 12, but not of IL-10. This result suggests that NKT cells are major components in the pathogenesis of HGA.

Effects of several suspending media on behavior of Toxoplasma gondii (보존액이 Toxoplasma gondii의 생존성에 미치는 영향)

  • Jin, Joo-Eun;Chung, Kyung-Tae;Lee, Woo-Won;Yang, Ju;Lee, Gang-Rog;Kim, Geun-Kyu
    • Korean Journal of Veterinary Service
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    • v.22 no.2
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    • pp.129-134
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    • 1999
  • A preservation test of Toxoplasma gondii tachyzoites for considerable time was tried to obtain simple and economical methods using various suspending media at $4^{\circ}C$ rather than serial passage of the parasite in mice. The preservation period was a term that the tachyzoites were detected from the peritoneal fluid of mice after inoculation of 2$\times$$10^5$ organisms preserved according to the lapse of time. The numbers of tachyzoite per 1\textrm{mm}^3$ of the peritoneal fluid with 2$m\ell$ of the saline solution taken in 4days after inoculation were presented as percentage in proportion to the control. The numbers. of tachyzoite per 1\textrm{mm}^3$ of the peritoneal fluid of the control were consisted of the average number of the tachyzoites of 10 mice inoculated with 2$\times$$10^5$ organisms by serial mouse passage. The tachyzoites could be preserved for 26 days in the suspending medium of saline solution at $4^{\circ}C$ Ringer's solution for 18 days, Hank's solution for 28 days, and egg-glycerine solution for 50 days.

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BONE FORMATION BY HUMAN ALVEOLAR BONE CELLS (사람 치조골세포를 이용한 골형성)

  • Choi, Byung-Ho;Park, Jin-Hyoung;Huh, Jin-Young;Oh, Jin-Rok
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.28 no.1
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    • pp.42-45
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    • 2002
  • Cultures of primary human alveolar bone-derived cells were established from alveolar bone chips obtained from normal individuals undergoing tooth extraction. These cells were expanded in vitro until passage 3 and used for the in vivo assays. Cells were loaded into transplantation vehicles, and transplanted subcutaneously into immunodeficient mice to study the capacities of human alveolar bone-derived cells to form bone in vivo. Transplants were harvested 12 weeks after transplantation and evaluated histologically. Of 10 human alveolar bone-derived cell transplants, two formed a bone-like tissue that featured osteocytes and mineral. Eight of the ten formed no osseous tissue. These results show that cells from normal human alveolar bone are capable of forming bone-like tissue when transplanted into immunodeficient mice.

Development of a New Gut-targeted Oral Typhoid Vaccine Ty21a Encasulated within Alginate Enteric Beads (알긴산 장용 비드에 봉입한 새로운 장 표적성 경구용 장티푸스 Ty21a 백신의 개발)

  • 장윤정;정성균;박동우;김희준;김기호
    • YAKHAK HOEJI
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    • v.45 no.5
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    • pp.536-544
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    • 2001
  • increase the viability of oral typhoid vaccine during the passage through the castro-intes-tidal tract, numerous attempts have been made including the vaccine coating. However problems such as high death rate during the coating process and its instability in the gastric juice still remain to be solved. In this study, the oral vaccine was made as the micro-enteric beads by adding Salmomella typhi Ty21a cells to sodium alginate solution and spraying onto calcium chloride solution (ionotropic relation method). The vaccine showed more than 90% of its original viability after treating it for 1 hour in the artificial gastric juice (37$^{\circ}C$, 300 rpm). The clearance rate of the Ty21a in the liver and spleen of the mice orally administrated with coated Ty21a was similar to that of the mice intraperitoneally administrated with uncoated Ty21a. The peripheral blood lymphocytes (PBL) isolated from the mice orally administered with this vaccine produced 15.5 fold higher specific IgA antibody titer than that from the control mice administerd with saline solution. furthermore, the mice treated with the coated Ty21a had higher survival rates (50~87%) than the control mice treated with saline solution (0~10%) in the intraperitoneal challenge test with wild type S. typhi Ty21a cells. These results suggest that the alginate-based coating technique is effective to protect live Ty21a from acidic environments, and produces better intestinal immune responses thereby providing a potentially excellent oral typhoid vaccine.

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Antidiarrheal Effect of LacteolTM-Loperamide Combination on Castor oil-induced Mice Model

  • Hwang, Se-Hee;Sung, Hee-Jin;Chung, Yong-Ho;Ryu, Jei-Man;Seong, Seung-Kyoo
    • Biomolecules & Therapeutics
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    • v.10 no.4
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    • pp.236-239
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    • 2002
  • The goal of this study was to evaluate the antidiarrheal efficacy of $Lacteol^{TM}$-loperamide combination against the mouse model of secretory diarrhea. Secretory dirrhea was induced in mice by p.o. administration of castor oil (0.3 ml). Antidirrheal effects of $Lacteol^{TM}$-loperamide combination were compared with each individual component. $Lacteol^{TM}$-loperamide combination was the most potent among these agents, eliminating diarrhea in 100% of mice at a dose 1360/4 mg/kg (Lacteol/loperamide, respectively). In this study, we also measured changes of bodyweight as another indicator of the dirrhea, based on the assumption that lower bodyweight loss represented reduced fecal passage. The bodyweight loss of $Lacteol^{TM}$-loperamide combination administered group was 4 times lower than that of vehicle control. These findings indicate that $Lacteol^{TM}$-loperamide combination may be more potent than individual component in its antidiarrheal action, so we are going to challenge this combination for further study and clinical evaluation.