• Title/Summary/Keyword: In vivo development

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Evaluation of different media for ex vivo porcine lung culture model

  • Yang, Myeon-Sik;Zhou, Zixiong;Khatun, Amina;Nazki, Salik;Jeong, Chang Gi;Kim, Won Il;Lee, Sang Myeong;Kang, Seog-Jin;Lim, Chae Woong;Kim, Bumseok
    • Korean Journal of Veterinary Service
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    • v.41 no.4
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    • pp.263-269
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    • 2018
  • Developing drugs targeting respiratory pathogen is essential to control respiratory diseases. Many experiments have been performed under in vivo situation. However, in vivo experiments have economical and ethical issues. The objective of this study was to determine the possibility of developing an ex vivo lung culture system with possible application for respiratory infection studies. After isolating lungs from naïve pigs, agarose-inflated lung tissues were prepared and sliced manually. These sliced lung tissues were then subsequently placed on 24-well plates. Eight different combinations of media were used to determine the optimum ex vivo lung culture condition. In addition, lung tissues were infected with porcine reproductive and respiratory syndrome (PRRS) virus at a titer of $1{\times}10^4\;TCID_{50}/mL$. Virus growth was confirmed by titration in MARC-145 cells at 2, 4, 6 days post infection (dpi). We found that ex vivo lung culture in physiological environment was not media specific based on histopathology and cytotoxicity. However, under virus-infected condition, thickened alveolar walls in the lung tissues and stable virus titers at 2, 4, 6 dpi were shown in F12K medium suggesting that it was useful for tissue maintenance and virus infection using PRRS virus infected lung tissues. The present study shows the possibility of using porcine ex vivo lung model for respiratory infection studies.

Antioxidant Activity of Essential Oils from Wedelia chinensis (Osbeck) in vitro and in vivo Lung Cancer Bearing C57BL/6 Mice

  • Manjamalai, A.;Grace, V.M. Berlin
    • Asian Pacific Journal of Cancer Prevention
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    • v.13 no.7
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    • pp.3065-3071
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    • 2012
  • Aim: The present investigation was to evaluate the effects of essential oils of Wedelia chinensis (Osbeck) on free radicals and in vivo antioxidant properties. Methods: Essential oils were extracted using hydro-distillation and compound analysis was performed by GC-MS analysis. Screening for inhibitory activity was conducted by DPPH and OH-scavenging assays. In addition an in vivo study was carried out in cell line implanted cancer bearing mice with assessment of levels of catalase, superoxide dismutase, glutathione peroxidase, lipid peroxidation, nitric oxide and reduced glutathione. Finally, lungs were dissected out for histopathology study of metastasis. Results: GC-MS analysis revealed the presence of carvocrol and trans-caryophyllene as the major compounds with 96% comparison with the Wilily and NBS libraries. The essential oil exhibited significant inhibition in DPPH free radical formation. Whereas reducing power and hydroxyl radical scavenging activity are dose dependent. When compared with the standard, it was found that the essential oil has more or less equal activity in scavenging free radicals produced. In the animal studies, the level of antioxidant enzymes catalase, superoxide dismutase and glutathione peroxidase, as well as glutathione, were found to be increased in treated groups whereas lipid peroxidation and nitric oxide were reduced. Histopathology report also shows that the essential oil has a significant combating effect against cancer development. Conclusion: In all the in vitro assays, a significant correlation existed between the concentrations of the essential oil and percentage inhibition of free radicals. The in vivo studies also has shown a very good antioxidant property for the essential oil during cancer development. From, these results the essential oil can be recommended for treating disease related to free radicals and to prevent cancer development.

Recent advances in intravital microscopy for investigation of dynamic cellular behavior in vivo

  • Choo, Yeon Woong;Jeong, Juhee;Jung, Keehoon
    • BMB Reports
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    • v.53 no.7
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    • pp.357-366
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    • 2020
  • Currently, most biological research relies on conventional experimental techniques that allow only static analyses at certain time points in vitro or ex vivo. However, if one could visualize cellular dynamics in living organisms, that would provide a unique opportunity to study key biological phenomena in vivo. Intravital microscopy (IVM) encompasses diverse optical systems for direct viewing of objects, including biological structures and individual cells in live animals. With the current development of devices and techniques, IVM addresses important questions in various fields of biological and biomedical sciences. In this mini-review, we provide a general introduction to IVM and examples of recent applications in the field of immunology, oncology, and vascular biology. We also introduce an advanced type of IVM, dubbed real-time IVM, equipped with video-rate resonant scanning. Since the realt-ime IVM can render cellular dynamics with high temporal resolution in vivo, it allows visualization and analysis of rapid biological processes.

Functional Imaging of the Multidrug Resistance In Vivo (기능적 영상술을 이용한 다약제 내성의 체내 진단)

  • Lee, Jea-Tae
    • 대한핵의학회:학술대회논문집
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    • 2001.05a
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    • pp.66-75
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    • 2001
  • Although diverse mechanisms are involved in multidrug resistance for chemotherapeutic drugs, the development of cellular P-glycoprotein(Pgp) and multidrug-resistance associated protein (MRP) are important factors in the chemotherapy failure to cancer. Various detection assays provide information about the presence of drug efflux pumps at the mRNA and protein levels. However these methods do not yield information about dynamic function of Pgp and MRP un vivo. Single photon emission tomography (SPECT) and positron emission tomography (PET) are available for the detection of Pgp and MRP-mediated transport. $^{99m}Tc$-sestaMIBl and other $^{99m}Tc$-radiopharmaceuticals are substrates for Pgp and MRP, and have been used in clinical studies for tumor imaging, and to visualize blockade of Pgp-mediated transport after modulation of Pgp pump. Colchicine, verapamil and daunorubicin labeled with $^{11}C$ have been evaluated for the quantification of Pgp-mediated transport with PET in vivo and reported to be feasible substrates with which to image Pgp function in tumors. Leukotrienes are specific substrates for MRP and N-$[^{11}C]$acetyl-leukotriene E4 provides an opportunity to study MRP function non-invasively in vivo. Results obtained from recent publications are reviewed to confirm the feasibility of using SPECT and PET to study the functionality of MDR transporters in vivo.

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Development of Glioblastoma In Vivo Model for the Research of Brain Cancer Diagnosis and Therapy (뇌암 진단 및 치료 연구를 위한 교모세포종 동물모델 개발)

  • Kang, Seonghee;Kang, Bosun
    • Journal of the Korean Society of Radiology
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    • v.8 no.7
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    • pp.389-395
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    • 2014
  • The research was carried out to develop a animal model of malignant brain tumor for the researches in glioblastoma multiform (GBM) diagnosis and therapy. C6 cells were transplanted into the right striatum of SD rat using stereotactic instrument for the development. The developed animal model was verified by MRI and H&E stain assay of anatomicohistological examination. The MRI observations showed that the tumor developed at the injection site at the 7 days after glioblastoma inoculation. At 14 days post inoculation, the tumor grew to a large volume occupying almost a half of the right cerebral hemisphere. It was confirmed that the expression of excessive mitosis and pleomorphism in anatomicohistological examination. The developed animal model must be necessary and useful tool for the in vivo level research in the development of the new modality for the diagnosis and therapy of brain cancer.

A STUDY ON A COMPARISON BETWEEN IN-VIVO AND IN-VITRO PHOTOTOXICITY TEST (IN-VIVO와 IN-VITRO에서의 광독성 시험법의 비교에 대한 연구)

  • Lee, Ho;Koh, Jae-Sook;Park, Won-Jae
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.19 no.1
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    • pp.57-76
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    • 1993
  • Phototoxicity is a complex phenomenon which may involve photochemical reaction and biological response mechanism. This complexicity and iii mal protecting tendency has led to the development of various in-vitro approaches as sensitive, alternative test to the in-vivo phototoxicity test. In this study, we investigated not only the sensitivity of two microorganism, (C. albicans and 5. typhimurium TA 98 about UV) but also a correlation between in-vitro and in-vivo phototoxicity test using UV A and 1 Furthermore, we studied the effect of irradiation method which were as follows 1) irradiate to material and microorganism, simultaneously 2) irradiate to only material 3) irradiate to material and microorganism, respectively In each irradiation method, it showed no significant difference, However we were able to observe the more sensitive phototoxicity in S. typhimurium TA 98 than C. albicans, and the results of in-vitro test using 5. typhimurium TA 98 had a good correlation with those of in-vivo test.

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Development of Dose Verification Method for In vivo Dosimetry in External Radiotherapy (방사선치료에서 투과선량을 이용한 체내선량 검증프로그램 개발)

  • Hwang, Ui-Jung;Baek, Tae Seong;Yoon, Myonggeun
    • Progress in Medical Physics
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    • v.25 no.1
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    • pp.23-30
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    • 2014
  • The purpose of this study is to evaluate the developed dose verification program for in vivo dosimetry based on transit dose in radiotherapy. Five intensity modulated radiotherapy (IMRT) plans of lung cancer patients were used in the irradiation of a homogeneous solid water phantom and anthropomorphic phantom. Transit dose distribution was measured using electronic portal imaging device (EPID) and used for the calculation of in vivo dose in patient. The average passing rate compared with treatment planning system based on a gamma index with a 3% dose and a 3 mm distance-to-dose agreement tolerance limit was 95% for the in vivo dose with the homogeneous phantom, but was reduced to 81.8% for the in vivo dose with the anthropomorphic phantom. This feasibility study suggested that transit dose-based in vivo dosimetry can provide information about the actual dose delivery to patients in the treatment room.

Effect of Culture Media and Co-culture with Bovine and Rabbit Oviductal Epithelial Cells on In Vitro Development of Rabbit Embryos (토끼 수정란의 체외발달에 미치는 배양액 및 소와 토끼의 난관상피세포들과의 공배양 효과)

  • 노규진;이효종;송상현;윤희준;박충생
    • Korean Journal of Animal Reproduction
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    • v.18 no.1
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    • pp.39-46
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    • 1994
  • This experiment was carried out to develop an in vitro culture system for rabbit embryos. The zygotes or 2-cell embryos were collected from the oviducts of the superovulated and mated does with D-PBS/10% FCS at 24 hours after hCG injection. The in vitro developmental rate of blastocyst formation and the number of nuclei in the embryos were examined under the following treatments; 1) TCM-199 with 10% FCS, 2) EBSS with 10% FCS, 3) rabbit vitreous humor(VH), 4) TCM-199 with 10% FCS+BOEC, 5) TCM-199 with 10% FCS+ROEC, 6) EBSS with 10% FCS+BOEC and 7) EBSS with 10% FCS+ROEC. For a comparative study of in vivo and in vitro development, the fresh blastocysts, which were developed in vivo for 96 hours after hCG injection, were collected from the uterus and their numbers of nuclei were counted. 1. The zygotes or 2-cell embryos developed to the blastocyst stage in TCM-199, EBSS and VH at the rates of 93, 92 and 89%, respectively. 2. The higher developmental rates 95~98% of blastocyst formation was achieved when the embryos were co-cultured with a monolayer of bovine or rabbit oviductal epithelial cells in TCM-199 or EBSS. No significant difference in developmental rates was shown between bovine and rabbit oviductal epithelial cells. 3. In a comparative study of in vivo and in vitro development, the total numbers of nuclei were significantly less in the in vitro cultured embryos(104~224) than the in vivo developed embryos(1, 0090 at 96 hours after hCG injectin. 4. The mean cell cycle numbers in the embryos cultured for 72 hours in TCM-199 with 10% FCS, EBSS with 10% FCS, TCM-199 with 10% FCS+BOEC, TCM-199 with 10% FCS+ROEC, EBSS with 10% FCS+BOEC and in vivo was 7.38, 6.63, 7.76, 7.69, 7.01 and 9.92, respectively. From these results, it can be suggested the optimal culture system for in vitro culture of rabbit embryos is a co-culture system with bovine or rabbit oviductal epithelial cells in TCM-199 with 10% FCS. Considering the significant reduction in total numbers of nuclei in the in vitro cultured embryos, the advanced research on development of in vitro culture system for rabbit embryos is expected.

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Ex vivo Cytotoxicity of the Bacillus thuringiensis Cry4B δ-Endotoxin to Isolated Midguts of Aedes aegypti Larvae

  • Barusrux, Sahawat;Sramala, Issara;Katzenmeier, Gerd;Bunyaratvej, Ahnond;Panyim, Sakol;Angsuthanasombat, Chanan
    • BMB Reports
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    • v.36 no.3
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    • pp.294-298
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    • 2003
  • The pathological effect of the Bacillus thuringiensis Cry $\delta$-endotoxins on susceptible insect larvae had extensive damage on the midgut epithelial cells. In this study, an ex vivo assay was devised for assessing the insecticidal potency of the cloned Cry4B mosquito-larvicidal protein that is expressed in Escherichia coli. Determination of toxicity was carried out by using a cell viability assay on the midguts that were dissected from 5-day old Aedes aegypti mosquito larvae. After incubation with the toxin proteins, the number of viable epithelial cells was determined photometrically by monitoring the quantity of the bioreduced formazan product at 490 nm. The results showed that the 65-kDa trypsin-activated Cry4B toxin exhibited toxic potency ca. 3.5 times higher than the 130-kDa Cry4B protoxin. However, the trypsin-treated products of the non-bioactive Cry4B mutant (R158A) and the lepidopteran-specific Cry1Aa toxin displayed relatively no ex vivo activity on the mosquito-larval midguts. The ex vivo cytotoxicity studies presented here confirms data that was obtained in bioassays.

Comparison of the Digestibility of Grain and Forage by Sheep, Red and Fallow Deer

  • Ru, Y.J.;Glatz, P.C.;Miao, Z.H.;Swanson, K.;Falkenberg, S.;Wyatt, S.
    • Asian-Australasian Journal of Animal Sciences
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    • v.15 no.6
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    • pp.800-805
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    • 2002
  • Two experiments were conducted to compare digestibility of 12 diets in sheep, red and fallow deer. No differences (p>0.05) between sheep, red and fallow deer in digestibility of dry matter, organic matter and digestible energy content for all diets were found except for the sorghum diet and medic hay. Sheep and fallow deer digested the sorghum diet better than red deer. An in vitro study showed that sheep had a lower in vitro dry matter digestibility and digestible energy content than both red and fallow deer, with a significant interaction between animal species and feed ingredient. Deer digested straws and hays better (p<0.05) than sheep. In vitro digestibility was lower (p<0.05) than in vivo digestibility, but significantly correlated with in vivo digestibility for red and fallow deer. The in vitro method for digestibility estimation has potential as a rapid feed evaluation system for deer, but needs further validation.