• 제목/요약/키워드: Inoculation methods

검색결과 217건 처리시간 0.028초

오제스키병의 생체 조기진단을 위한 면역세포화학, In situ hybridization 및 전자현미경적 연구 (Immunocytochemistry, In situ hybridization and electron microscopy for early diagnosis of Aujeszky's in living pigs)

  • 문운경;김순복;서정향;송근석;노환국
    • 대한수의학회지
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    • 제36권4호
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    • pp.845-858
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    • 1996
  • The purpose of this study was to establish early diagnostic methods for the detection of Aujeszky's disease viral antigens and nucleic acid in nasal cells, and buffy coats from experimentally infected living pigs by a combination of immunocytochemistry, in situ hybridization with digoxigenin(DIG)-labled probe and electron microscopy. Forty days old piglets were inoculated intranasally with $10^{7.0}TCID_{50}$ of Aujeszky's disease virus (ADV, NYJ-1-87 strain). The viral antigens and nucleic acid of ADV were detected in nasal cells, and buffy coat for 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopical method. The results were compared with conventional methods such as a porcine Aujeszky's disease serodiagnostic(PAD) kit, neutralization test(NT) and virus isolation. 1. The viral antigens, nucleic acids and capsids of ADV were detected in nasal cells, buffy coats from 3 days to 20 days after inoculation by immunocytochemistry, in situ hybridization with DIG-labeled probe and electron microscopy, respectively. 2. When viral antigens were detected by the immunocytochemical technique, a diffuse brown deposit was observed in the nucleus and cytoplasm of nasal cells, buffy coats and PK-15 cells under a microscope. 3. DIG-labeled DNA probe was prepared by amplification of conserved sequence of recombinant ADV-gp50 clone with polymerase chain reacction. When ADV-DNA was detected by ISH with DIG-labeled probe, purplish blue pigmentation were observed in the nuclei and cytoplasms of ADV-infected cells under a microscope. Positive signals were observed in nasal cells and in the buffy coat and PK-15 cells at the first day after inoculation. 4. Where ADV-capsids were detected by transmission electron microscopical method, aggregation of capsids was observed in the nuclei and cytoplasms of nasal cells, buffy coats and PK-15 cells. The results suggested that these methods were considered as the highly sensitive and reliable tools for rapid and confirmative diagnosis of Aujeszky's disease in living pigs.

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Neuroradiology 연구를 위한 VX2 세포를 이용한 토끼 뇌종양 모델 제작과 MRI를 이용한 검증 (Development of Rabbit Brain Tumor Model Using VX2 Cells and Verification with the MRI in Neuroradiologic Research)

  • 김용우;최선희;김학진
    • 대한영상의학회지
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    • 제84권2호
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    • pp.441-453
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    • 2023
  • 목적 네 가지 방법으로 토끼의 뇌에 VX2 세포를 이식하여 뇌종양의 생성 유무와 위치, 체적을 분석하고자 한다. 대상과 방법 공여 토끼를 희생하여 얻은 VX2 세포 현탁액을 1) 당일 이식, 2) 냉장 보관 7일 후 이식, 3) 초저온 냉동보관 7일 후 이식, 그리고 4) 공여 토끼를 희생하지 않고 조직생검 기구를 이용하여 VX2 세포현탁액을 얻은 후 당일 이식하였다. 뇌 이식에는 정위 기구를 사용하였으며 하방 레버를 이용하여 5 mm 진입하여 VX2 세포 현탁액을 주입하였다. 10일 경과 후 MR imaging을 실시하였으며 다음 날 뇌 조직을 얻어 조직검사를 실시하여 MR 영상과 현미경 검사상 뇌종양의 생성 유무와 위치, 체적 등을 살펴보았다. 결과 VX2 세포 현탁액을 당일 이식한 토끼(n = 18)에서는 17마리(94.4%)에서 뇌종양이 생겼다(평균 체적은 106.32 mm3). 냉장 보관 후 7일에 세포 현탁액을 만들어 뇌 이식한 토끼(n = 5)에서는 1마리(20%)에서 뇌종양이 생겼으나 현미경으로만 확인되었다. 초저온 냉동 보관 후 7일에 세포 현탁액을 만들어 뇌 이식한 토끼는 5마리에서는 3마리(60%)에서 뇌종양이 생겼고 현미경으로만 확인되었다. 생검 횟수를 10회 실시한 후 이식한 3마리에서는 뇌종양이 생기지 않았고, 20회 실시한 후 이식한 2마리에서는 모두 뇌종양이 생겼으며 MR 영상과 조직검사에서 확인되었다(평균 체적 38.71 mm3). 네 가지 방법으로 발생한 종양의 위치는 superficial cortex가 대부분이었다. 결론 VX2 세포를 이식하여 토끼의 뇌종양 모델을 제작하는 기법은 쉽고 재현성은 다양했다. 이 모델은 뇌종양에 대한 영상, 치료 방법, 중재 시술 및 약물 전달 등 다양한 연구에 활용될 수 있을 것이다. 연구자의 처한 다양한 환경에 따라 다양한 방법으로 VX2 세포를 이식할 수 있을 것으로 생각된다.

건강증진학교 모형개발 - 대형 보건소 활동을 중심으로 - (Modeling of Health-Promoting School - Based on Health Center of University -)

  • 하은희;조희숙;윤영옥;강명근
    • 한국학교보건학회지
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    • 제14권1호
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    • pp.83-93
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    • 2001
  • Objectives: The purpose of this study is to develop a health-promoting program centered on the university-health center and to provide a developed program for students and faculties. Methods: A survey was conducted of 719 female students and 238 faculties concerning their health-promoting programs. Based on the results of the survey, we developed programs that could be applied to students and faculties. After operating the programs, we developed further models by evaluating the effectiveness and satisfaction. Conclusions: We selected a comprehensive health-promoting program that included weight control, preventive inoculation, sex education and control of disease based on repsective needs. We also applied programs developed through collaboration with other departments from September 1998 to August, 1999. Users of the preventive inoculation-program increased by one point five during the year. Also, participants were generally satisfied with the weight control program and succeeded in reducing weight. We hope that we can set up this program and expand it by developing various other projects in Korea.

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Antifungal Activity of Nor-securinine Against Some Phytopathogenic Fungi

  • Sahni, Sangita;Maurya, S.;Singh, U.P.;Singh, A.K.;Singh, V.P.;Pandey, V.B.
    • Mycobiology
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    • 제33권2호
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    • pp.97-103
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    • 2005
  • Crude extracts and active principles from medicinal plants have shown potential role in controlling plant diseases in glasshouses as well as in fields as one of the safest and ecofriendly methods. The effect of nor-securinine (an alkaloid) isolated from Phyllanthus amarus has been seen against spore germination of some fungi (Alternaria brassicae, A. solani, Curvularia pennisetti, Curvularia sp., Erysiphe pisi, Helminthosporium frumentacei) as well as pea powdery mildew (Erysiphe pisi) under glasshouse conditions. The sensitivity of fungi to nor-securinine varied considerably. Nor-securinine was effective against most of the fungi. H. frumentacei was more sensitive even at the lowest concentration ($1,000\;{\mu}g/ml$). Likewise conidia of E. pisi were also inhibited in partially or completely appressorium formation. Pre-inoculation treatment showed greater efficacy than post-inoculation in inhibiting powdery mildew development on pea plants in a glasshouse. Maximum inhibition occurred at $2000\;{\mu}g/ml$.

Natural Variation in Virulence of Acidovorax citrulli Isolates That Cause Bacterial Fruit Blotch in Watermelon, Depending on Infection Routes

  • Song, Yu-Rim;Hwang, In Sun;Oh, Chang-Sik
    • The Plant Pathology Journal
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    • 제36권1호
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    • pp.29-42
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    • 2020
  • Acidovorax citrulli causes bacterial fruit blotch in Cucurbitaceae, including watermelon. Although A. citrulli is a seed-borne pathogen, it can cause diverse symptoms in other plant organs like leaves, stems and fruits. To determine the infection routes of A. citrulli, we examined the virulence of six isolates (Ac0, Ac1, Ac2, Ac4, Ac8, and Ac11) on watermelon using several inoculation methods. Among six isolates, DNA polymorphism reveals that three isolates Ac0, Ac1, and Ac4 belong to Clonal Complex (CC) group II and the others do CC group I. Ac0, Ac4, and Ac8 isolates efficiently infected seeds during germination in soil, and Ac0 and Ac4 also infected the roots of watermelon seedlings wounded prior to inoculation. Infection through leaves was successful only by three isolates belonging to CC group II, and two of these also infected the mature watermelon fruits. Ac2 did not cause the disease in all assays. Interestingly, three putative type III effectors (Aave_2166, Aave_2708, and Aave_3062) with intact forms were only found in CC group II. Overall, our results indicate that A. citrulli can infect watermelons through diverse routes, and the CC grouping of A. citrulli was only correlated with virulence in leaf infection assays.

Xenografted Tumorigenesis in the oral vestibule of nude mice by Snail transfection: Histological and immunohistochemical study

  • Kim, Moon-Key;Lee, Eun-Ha;Kim, Jin;Yook, Jong-In;Cha, In-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • 제35권4호
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    • pp.199-204
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    • 2009
  • Purpose: The purpose of this study is to investigate the epithelial-mesenchymal transition (EMT) induced by Snail transcription factor and Snail-transfected in vivo tumors with histopathological features. Materials and methods: We induced in vivo xenografted tumorigenesis in the oral vestibules of nude mice by a Snail transfected HaCaT cell line and investigated morphological and immunohistochemical features in Snail expressive tumors. Results: We identified tumor masses in 14 out of 15 nude mice in the HaCaT-Snail cell inoculation group, but no tumors were present in any of the HaCaT cell inoculation group. Induced tumors showed features of poorly differentiated carcinoma with invasion to neighboring muscles and bones. The HaCaT-Snail tumors showed decreased expressions of E-cadherin and cytokeratin, but showed increased expressions of vimentin and N-cadherin. Discussion: The Snail transfected xenograft can improve productivity of malignant tumors, show various histopathological features including invasive growth, and aid in the investigation of tumor progression and the interaction with surrounding tissues.

넙치에서 방사선을 조사한 병원성 세균 오염사료 투여의 효과 (The Effect of Food Treated with Gamma Radiation after Inoculation with Pathogenic Bacteria in the Flounder (Paralichthys olivaceus))

  • 김세라;김성호
    • 한국임상수의학회지
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    • 제19권1호
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    • pp.7-13
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    • 2002
  • This study was examined the microbiological adequacy of fish feed treated with high-dose irradiation (5 kGy). 125 flounder (Paralichthys olivaceus) were grouped into 5 and then the fishes were fed the following feeds for 28 days: (1)standard feed; (2)standard feed, inoculated with Edwardsiella tarda ($1{\times}10^8-1{\times}10^9CFU/g$ of feed); (3)standard feed, inoculated with Vibrio anguilarum ($1{\times}10^8-1{\times}10^9CFIT/g$ of feed); (4)standard feed, inoculated with Streptococcus faecalis ($1{\times}10^8-1{\times}10^9CFU/g$ of feed); (5)standard feed, inoculated with Edwardsiella tarda, Vibrio anguillarum and Streptococcus faecalis, and then irradiated the mixed feed to 5 kGy. The flounders feed the mixed diet with Edwardsiella tarda, Wbrio anguillarum or Streptococcus faecalis inoculated feed were showed severe cumulative mortalities of 60, 48 and 52% respectively. The gross and histological changes were observed on the fishes. However, fishs fed with the feed of bacteria inoculation before irradiation demonstrated excellent protection against the bacteria-related disease. The results from experiments with bacteria inoculated feed indicated that the irradiation methods employed were capable of preventing contamination of the fishs with pathogenic bacteria.

마우스에 있어서 Dermatophilus 양 균에 의한 농양의 형태 병리 발생 (Morphopathogenesis o the abscess induced by Dermatophilus-like microorganism in mice)

  • 하창수;박응복
    • 대한수의학회지
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    • 제33권4호
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    • pp.633-647
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    • 1993
  • NC strain mice were inoculated intraperitoneally with Dermatophoilus-like microorganism strain W254 isolated from the sulfur granules of the swine tonsils. The development of the resulting abscesses in NC strain mice was investigated at regular time intervals by means of histological, histochemical and electron microscopical methods. The results obtained were as follows ; 1. The inoculated bacteria have settled in the subserous site of the peritoneal organ and provoked cellular response to form neutrophil, macrophage and connective tissue layers at the time of one week after inoculation, and thereafter were found to have formed typical actinomycotic abscess two weeks after inoculation. 2. The persistant abscesses were appeared to reveal close interaction between higher activity of organism and tissue reaction and contained hyaloid bodies within or outside bacterial clumps. 3. The hyaline materials were appeared to be composed of protein rich in hydroxyphenyl group by histochemical examination. The materials were presumed to be produced by the interaction between bacterial activity and host cell reaction. 4. The bacteria in the abscesses showed the various developmental stages of coccoid body, mulberry-like body tuber-shaped body and filamentous body, and it was confirmed that the mesosomal development was related to septation of the bacterial cells.

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Effects of Arugula Vermicompost on the Root-Knot Nematode (Meloidogyne javanica) and the Promotion of Resistance Genes in Tomato Plants

  • Rostami, Mahsa;Karegar, Akbar;Ghorbani, Abozar
    • The Plant Pathology Journal
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    • 제38권4호
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    • pp.261-271
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    • 2022
  • Root-knot nematodes are the most important plant-parasitic nematodes worldwide. Many efforts have been made to find non-chemical, risk-free, and environmentally friendly methods for nematode control. In this study, the effects of compost and vermicompost of arugula (Eruca sativa) on Meloidogyne javanica were investigated in three glasshouse experiments. In addition, the expression of the defense-related genes nonexpressor of pathogenesis-related 1 (NPR1) and lipoxygenase 1 (LOX1) was detected in tomato plants treated with vermicompost of arugula at 0, 2, 7, and 14 days after nematode inoculation. The result showed that the vermicompost of arugula significantly reduced the reproduction factor of the nematode by 54.4% to 70.5% in the three experiments and increased the dry weight of shoots of infected tomato plants. Gene expression analysis showed that LOX1 expression increased on the second and seventh day after nematode inoculation, while NPR1 expression decreased. The vermicompost of arugula showed stronger nematode inhibitory potential than the vermicompost of animal manure. The vermicompost of arugula is superior to arugula compost in suppressing the activity of M. javaniva and reducing its impact. It manipulates the expression of resistance genes and could induce systemic resistance against root-knot nematodes.

경피용 건조 B.C.G. 백신(Tokyo 172주) 접종 후 국소 반응과 투베르쿨린 양전율에 관한 연구 (Observation of Response to PPD Skin Test and Local Side Reactions at Multiple Inoculation Sites after Percutanous Inoculation with BCG Tokyo 172 Strain)

  • 이진수;손영모
    • Pediatric Infection and Vaccine
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    • 제7권2호
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    • pp.201-210
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    • 2000
  • 목 적 : 관침관을 이용한 분산난자 경피용으로 접종하는 일본에서 제조한 Tokyo 172주 냉동건조 비시지 백신의 투베르쿨린 양성율과 피부국소 반응 및 부작용 빈도를 알아보기 위하여 본 조사를 시행하였다. 방 법 : 신생아, 영유아, 소아를 대상으로 영동세브란스병원과 7개 개인 소아과 의원에서 접종 후 3일, 4~6주, 36주, 48주째 국소 변화 및 림프절 종창 여부를 관찰하였다. 접종 후 4개월 후에 5TU 0.1mL를 전박 내측에 피내주사하여 48~72시간 후 판독하였다. 결 과 : 관찰기간 중 총 138명의 접종자 중 96명에서의 48주간 추적이 가능하였으며 5TU 투베르쿨린 반응검사 결과 경결 크기의 분포는 5mm 미만이 6명으로 6.3%, 10mm 이상이 67명으로 70.0%, 12mm 이상이 46명으로 47.9%였다. 접종자 모든 례에서 접종 부위 피부에 염증 반응이 관찰되었으며 대부분 4~6주 후에 구진과 작은 농포 및 궤양이 생겼다. 모든 례에서 8~12주 후에 가피를 형성한 후 증상이 사라졌으며 접종 후 48주에 관찰한 접종 부위 반흔 조사에 의하면 전체의 4.2%(4명)에서는 아무런 흔적도 발견할 수 없었고 약 70% 이상에서 10개 이상의 반흔이 관찰되었다. 접종 후 16주에 관찰된 5TU 투베르쿨린 검사의 경결 크기와 48주에 관찰한 접종부위 반흔의 개수와는 상관관계가 관찰되지 않았다. 접종 후 48주까지 추적한 96례 모두에서 액와부 림프절의 종창이나 화농성 염증 등의 이상 반응은 관찰되지 않았다. 결 론 : 본 연구에 사용된 관침관을 이용한 분산난자 경피용 접종법은 이상 반응이 관찰되지 않았으며, 통상적으로 피내 접종 후에 초래되는 어깨 부위의 작은 흉터는 한 례에서도 관찰되지 않았다. 모든 례에서 접종 후 48주 후가 되면 희미한 접종 자국만이 관찰되어 기존의 피내 접종으로 인한 흉터의 우려에 대한 보호자의 만족도는 비교적 높았다.

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