• Title/Summary/Keyword: Inoculation methods

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

  • Moon, Oun-kyong;Kim, Soon-bok;Sur, Jung-hyang;Song, Geun-suk;Nho, Whan-gook
    • Korean Journal of Veterinary Research
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    • v.36 no.4
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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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Development of Rabbit Brain Tumor Model Using VX2 Cells and Verification with the MRI in Neuroradiologic Research (Neuroradiology 연구를 위한 VX2 세포를 이용한 토끼 뇌종양 모델 제작과 MRI를 이용한 검증)

  • Yong-Woo Kim;Seon Hee Choi;Hak Jin Kim
    • Journal of the Korean Society of Radiology
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    • v.84 no.2
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    • pp.441-453
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    • 2023
  • Purpose To evaluate the development, location, and volume of a VX2 carcinoma using four inoculation methods in a rabbit brain. Materials and Methods Inoculation of a VX2 cell suspension was performed 1) on the appointed day, 2) seven days after storing a VX2 carcinoma in a freezer or 3) seven days after storing a VX2 carcinoma in a deep freezer after sacrificing the donor rabbits. 4) Without sacrificing the rabbits, the VX2 cell suspension was obtained using a gun biopsy, inoculation was performed on the appointed day. MR imaging was performed 10 days after inoculation. Brain tissues were obtained the day after. The development, location, and volume of the tumor were evaluated. Results Seventeen of the 18 rabbits inoculated on the appointed day developed tumors (average tumor volume, 106.32 mm3). One of five inoculated seven days after storing the VX2 tumor in the freezer, and three of five inoculated seven days after storing the VX2 tumor in the deep freezer developed tumors. Inoculation with a VX2 cell suspension obtained with a gun biopsy from five rabbits revealed development of tumors in only two rabbits. The tumors mostly developed in the superficial cortex. Conclusion TVX2 rabbit brain tumor model is easy to develop and revealed variable reproducibility. This model can be applicable in radiologic imaging, treatment planning, interventional treatment and drug delivery research. VX2 cell can be successfully innoculated into the brain using variable methods under researcher's variable conditions.

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

  • Ha, Eunhee;Jol, Heuisug;Yun, Youngok;Kang, Myunggeun
    • Journal of the Korean Society of School Health
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    • v.14 no.1
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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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    • v.33 no.2
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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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    • v.36 no.1
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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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    • v.35 no.4
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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.

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

  • 김세라;김성호
    • Journal of Veterinary Clinics
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    • v.19 no.1
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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.

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

  • Ha, Chang-su;Bak, Ung-bok
    • Korean Journal of Veterinary Research
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    • v.33 no.4
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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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    • v.38 no.4
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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.

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

  • Lee, Jin Soo;Sohn, Young Mo
    • Pediatric Infection and Vaccine
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    • v.7 no.2
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    • pp.201-210
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    • 2000
  • Purpose : We observed response to PPD skin test and local side reactions among subjects who received inoculation with Tokyo 172 BCG strain by percutaneous method using multiple puncture device. Methods : 138 infants and young children were enrolled at Yongdong Severance Hospital and 7 private clinics. 5TU PPD skin test were performed at 4 months after inoculation. The local reactions at multiple puncture site were observed in 3 days, 4~6 weeks, 36 weeks, and 48 weeks after inoculations and physical check up was done for evaluation of lymphadenopathy. Results : During 48 weeks of observation period, 96 subjects among 138 who were enrolled were followed up completely with records of PPD skin test and observation of local side reactions, presenting with the photos. The size of the induration after 48 hours of PPD skin test, was less than 5mm in six subjects(6.3%), greater than 10mm in sixty seven subjects(70.0%) and greater than 12mm in forty six subjects(47.9%). All subjects showed inflammatory reaction and pustules at multiple puncture sites and only just small papules, ulcer and pustules remained 4-6 weeks later. Eight to twelve weeks later, all local inflammatory skin reactions disappeared with remaining crust. After 48 weeks, 4(4.2%) subjects showed no scar with only faint stain on the puncture site. More than 70% of subjects showed more than 10 faint pin-point scars on the sites. However, the size of scar was clearly smaller compared to that of intradermal inoculation. There were no cases of lymphadenopathy. Conclusion : We observed good immune response to 5TU PPD skin test among the infant and young children who were immunized with percutanous inoculation of Tokyo 172 BCG strain. We could not find any severe local scar at inoculation sites. A degree of satisfaction of the parents whose children received the percutaneous injection was very high.

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