• Title/Summary/Keyword: In vivo

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Post-Thaw Cryosurvival of Bovine Embryos Produced In Vitro and In Vivo after Controlled Freezing

  • Cho, Sang-Rae;Choi, Sun-Ho;Choe, Chang-Yong;Lee, Poong-Yeon;Son, Jun-Kyu;Kim, Jae-Bum;Kim, Sung-Jae;Kim, Hyun-Jong;Shin, Seung-Oh;Son, Dong-Soo
    • Journal of Embryo Transfer
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    • v.24 no.4
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    • pp.253-257
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    • 2009
  • To enhance the embryo preservation technology and better application of embryo transfer technique to the field (dairy science or animal reproduction. etc.), we examined the viabilities of bovine embryos produced in vitro and in vivo after cryopreservation according to their developmental stage and thawing temperature. Bovine embryos from in vivo/vitro fertilization (Hanwoo) were examined at day 7, 8, and 9. Survival rates and total cell numbers of in vivo fertilized embryos were as follows: morulae 68.8% and $67\;{\pm}\;6.0$; blastocysts 80.5% and $120\;{\pm}\;10$; expanded blastocysts 77.4% and $138\;{\pm}\;9.7$, respectively. Rates of embryo development for blastocysts and expanded blastocysts after thawing were significantly higher than that of morula stage embryos (p<0.05). While survival rates of in vitro fertilized embryos according to developmental stage showed no significant difference among groups (morula 67.9%; blastocyst 74.3%; and expanded blastocyst 79.4%), total cell numbers were significantly lower than those of other groups (morula $64\;{\pm}\;5.9$; blastocyst $116\;{\pm}\;8.7$; and expanded blastocyst $135\;{\pm}\;9.1$) For the viability according to thawing temperature, survival rate was higher in $37^{\circ}C$.

Assessment of Risks and Benefits of Using Antibiotics Resistance Genes in Mesenchymal Stem Cell-Based Ex-Vivo Therapy

  • Narayan Bashyal;Young Jun Lee;Jin-Hwa Jung;Min Gyeong Kim;Kwang-Wook Lee;Woo Sup Hwang;Sung-Soo Kim;Da-Young Chang;Haeyoung, Suh-Kim
    • International Journal of Stem Cells
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    • v.16 no.4
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    • pp.438-447
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    • 2023
  • Recently, ex-vivo gene therapy has emerged as a promising approach to enhance the therapeutic potential of mesenchymal stem cells (MSCs) by introducing functional genes in vitro. Here, we explored the need of using selection markers to increase the gene delivery efficiency and evaluated the potential risks associated with their use in the manufacturing process. We used MSCs/CD that carry the cytosine deaminase gene (CD) as a therapeutic gene and a puromycin resistance gene (PuroR) as a selection marker. We evaluated the correlation between the therapeutic efficacy and the purity of therapeutic MSCs/CD by examining their anti-cancer effect on co-cultured U87/GFP cells. To simulate in vivo horizontal transfer of the PuroR gene in vivo, we generated a puromycin-resistant E. coli (E. coli/PuroR) by introducing the PuroR gene and assessed its responsiveness to various antibiotics. We found that the anti-cancer effect of MSCs/CD was directly proportional to their purity, suggesting the crucial role of the PuroR gene in eliminating impure unmodified MSCs and enhancing the purity of MSCs/CD during the manufacturing process. Additionally, we found that clinically available antibiotics were effective in inhibiting the growth of hypothetical microorganism, E. coli/PuroR. In summary, our study highlights the potential benefits of using the PuroR gene as a selection marker to enhance the purity and efficacy of therapeutic cells in MSC-based gene therapy. Furthermore, our study suggests that the potential risk of horizontal transfer of antibiotics resistance genes in vivo can be effectively managed by clinically available antibiotics.

In Vivo Three-dimensional Motion Analysis of the Shoulder Joint During Internal and External Rotation at 90 Degrees of Abduction, using wide Gantry MRI.

  • Koishi, Hayato;Goto, Akira;Yoshikawa, Hideki;Sugamoto, Kazuomi
    • The Academic Congress of Korean Shoulder and Elbow Society
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    • 2009.03a
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    • pp.175-175
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    • 2009
  • Despite its importance for the understanding of joint kinematics in vivo, there has been few studies about shoulder joints. The purpose of this study is to analyze the glenohumeral joint during internal and external rotation at 90 degrees of abduction using in vivo noninvasive motion analysis system. MRI was performed for the following seven positions from maximum internal rotation to maximum external rotation at intervals of 30 degrees. We used 3D-gradient echo sequencing (TR: 12 ms, TE: 5.8 ms, 0.8 mm-slice thickness). Our method is based on matching three-dimensional MR images by the similarity of the image intensity. We analyzed the in vivo three-dimensional motions of the glenohumeral and scapulothoracic joint during this motion. In scapla plane, the mean rotation angle of the glenohumeral join was 105.5 degrees ($SD{\pm}39.0^{\circ}$). The mean rotation angle of the scapulothracic joint was 27.5 degrees ($SD\;{\pm}\;7.7^{\circ}$). The contribution ratio is almost 3.8:1 of glenohumeral and scapulothracic joint respectively.

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Anticardiovascular Diseases Effects of Fermented Garlic and Fermented Chitosan

  • Kim, Hyun-Kyoung;Lee, Jeong-Hun
    • International Journal of Advanced Culture Technology
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    • v.6 no.4
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    • pp.109-115
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    • 2018
  • Garlic is a medicinal plant used throughout the world for its anti-inflammatory, antioxidant, and antiplatelet activities. Chitosan is a natural polysaccharide obtained from chitin, and derivatives of chitosan have been shown to inhibit platelet aggregation and adhesion. We hypothesized that fermented preparations of these products may possess stronger antiplatelet effects than the non-fermented forms owing to the increased bioavailability of the bioactive compounds produced during fermentation. Therefore, we compared these compounds via in vitro and ex vivo platelet aggregation assays by using standard light transmission aggregometry and ex vivo granule secretions from rat platelets. We found that fermented preparations exerted more potent and significant inhibition of platelet aggregation both in vitro and ex vivo. Likewise, ATP release from dense granules of platelets was also significantly inhibited in fermented preparation-treated rat platelets compared to that in non-fermented preparation-treated ones. We concluded that fermented preparations exerted more potent effects on platelet function both in vitro and ex vivo, possibly as a result of the increased bioavailability of active compounds produced during fermentation. We therefore suggest that fermented products may be potent therapeutics against platelet-related CVDs and can be used as antiplatelet and antithrombotic agents.

Inhibitory Effects of Phenolic Alkaloids of Menispermum Dauricum on Gastric Cancer in Vivo

  • Zhang, Hong-Feng;Wu, Di;Du, Jian-Kuo;Zhang, Yan;Su, Yun-Ming
    • Asian Pacific Journal of Cancer Prevention
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    • v.15 no.24
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    • pp.10825-10830
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    • 2015
  • The present study was conducted to investigate effects and mechanisms of action of phenolic alkaloids of Menispermum dauricum (PAMD) on gastric cancer in vivo. In vitro, cell apoptosis of human gastric cancer cell line SGC-7901 was observed using fluorescence staining. In vivo, a mice model was constructed to observe tumor growth with different doses. Cell apoptosis was examined using flow cytometry and K-RAS protein expression using Western blotting. The mRNA expression of P53, BCL-2, BAX, CASPASE-3, K-RAS was examined by real-time PCR. PAMD significantly suppressed tumor growth in the xenograft model of gastric cancer in a dose-dependent manner (p<0.01). Functionally, PAMD promoted cell apoptosis of the SGC-7901 cells and significantly increased the rate of cell apoptosis of gastric tumor cells (p<0.05). Mechanically, PAMD inhibited the expression of oncogenic K-RAS both at the mRNA and protein levels. In addition, PAMD affected the mRNA expression of the cell apoptosis-related genes (P53, BCL-2, BAX, CASPASE-3). PAMD could suppress gastric tumor growth in vivo, possibly through inhibiting oncogenic K-RAS, and induce cell apoptosis possibly by targeting the cell apoptosis-related genes of P53, BCL-2, BAX, CASPASE-3.

Optimization of In Vivo Stickiness Evaluation for Cosmetic Creams Using Texture Analyzer (Texture Analyzer (TA)를 이용한 화장품 크림의 In Vivo 끈적임 평가법의 최적화)

  • Ryoo, Joo-Yeon;Bae, Jung-Eun;Kang, Nae-Gyu
    • Journal of the Society of Cosmetic Scientists of Korea
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    • v.46 no.4
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    • pp.371-382
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    • 2020
  • There have been continuous attempts to quantify sensory attributes of cosmetic products by measuring relevant physical properties. The most representative method to evaluate stickiness is to measure axial force using texture analyzer. Stickiness is known to correlate with AUC which abbreviates area under curve in the obtained axial force curve as a function of time. Recently, Normandie University research group developed in vivo stickiness evaluation method considering the characteristics of skin along with established evaluation method[8]. Based on the study, we tried to optimize in vivo stickiness evaluation method especially for cosmetic creams. The experiment was carried out on 5 different facial creams products by changing the amount and the times of rolling of creams, and the shape and material of probes. Based on the results of the sensory evaluation, the most consistent conditions were established as the optimal evaluation method. As a result, applying 70 μL of cream and rubbing 10 times for 7 s inside the 3.4 cm circle were judged to be suitable. As for the probes, spherical metallic probe was more proper due to its reproducibility. We conducted the settled method on 10 subjects to check its validity. Although the absolute values of AUC differed depending on the individuals, the AUC values were all ranked the same. Finally, for the standardization of stickiness of AUC, polyvinylpyrrolidone (PVP) was set as a reference material and we measured AUC of its aqueous solution by changing concentration. Then, the degree of stickiness recognition for 5 different creams was surveyed to check the correlation between AUC and stickiness.

Chest Wall Thickness Measurement of the LLNL Phantom for Ce Detectors (Ce 검출기를위한 LLNL 팬텀의가슴벽두께측정)

  • Lee, Tae-Young;Lee, Jong-Il;Chang, Si-Young
    • Journal of Radiation Protection and Research
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    • v.25 no.1
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    • pp.3-9
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    • 2000
  • The Korea Atomic Energy Research Institute acquired the Lawrence Livermore National Laboratory phantom for calibration of germanium detectors used for in vivo measurement of radionuclides deposited in human lungs. The manufacturer inscribed concentric circles as a phoswich detector positioning guides on the phantom's torso plate and its overlay plates, and provided the effective thickness of the chest wall for each plate measured at locations over the circles. However, since the germanium detectors are of different sizes, the areas considered for phoswich detectors were no. longer applicable for the locations of the germanium detectors on the phantom. Therefore, we re-evaluated the effective thickness of the phantom to determine if the manufacturer' s data are valid for germanium detectors in use for in vivo lung counting or if new values must be implemented. Differences no more than 3% in effective thickness were found between the germanium detector regions to be used at the Korea Atomic Energy Research Institute and the phoswich detector regions prescribed by the manufacturer.

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Effects of the ex-vivo Immunotherapy on the Mammary Gland Tumorigenesis Induced by 7, 12-dimethylbenz[a]anthracene(DMBA) in rats (7, 12-dimethylbenz[a]anthracene(DMBA) 투여에 의한 랫드 유선암 모델에서 ex-vivo 면역치료 효과)

  • 정자영;김옥희;이영순
    • Toxicological Research
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    • v.14 no.4
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    • pp.465-474
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    • 1998
  • This study was examined on the effect of ex-vivo immunotherapy in 7, 12-dimethylbenz[a]anthracene (DMBA)-induced rat mammary carcinogenesis. Sprague-Dawley female 40 rats were divided into Jour groups. As a positive control, Group I was intubated with DMBA, 5 mg /100 g body weight and single dose, at experimental onset. Group II was treated ex-vivo immunotherapy with polyinosinic-polycytidylic acid (Poly I : C) and Group III was treated with Interleukin-2 (IL-2). Group IV was negative control. All rats were sacrificed at 16 weeks after DMBA intubation. Mammary gland wet weight, dry fat free tissue weight, incidence of tumor, and the number of lobules, alveolar buds, terminal end buds, and terminal ducts were examined. Morphological changes of the mammary gland after treated with DMBA were analyzed by whole mount and histopathological method. As results, the induced mammary tumors of Group I, II and III were 60%, 33% and 0%, respectively. Histopathological types of induced-mammary tumors were adenoma, adenocarcinoma and carcinosarcoma. In analysis of the whole mount method, the number of the terminal end buds, terminal ducts and lobules were significantly lower in Group II (p<0.01) and III (p<0.01) than DMBA alone treated Group I. In microscopic observation, hyperplastic alveolar nodules were significantly lower in Group III than Group I (p<0.01). In conclusion, IL-2 had strong inhibitory effect on the mammary gland tumorigenesis induced by DMBA in rats. Whole mount method may be a useful technique to assess the mammary carcinogenesis. Moreover, hyperplastic alveolar nodules were very sensitive parameter to assess the mammary carcinogenesis.

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Identification of Enterococcus faecalis antigens specifically expressed in vivo

  • Lee, Seok-Woo;Shet, Uttom K.;Park, Sang-Won;Lim, Hyun-Pil;Yun, Kwi-Dug;Kang, Seong Soo;Kim, Se Eun
    • Restorative Dentistry and Endodontics
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    • v.40 no.4
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    • pp.306-313
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    • 2015
  • Objectives: Molecular mechanism of the pathogenicity of Enterococcus faecalis (E. faecalis), a suspected endodontic pathogen, has not yet been adequately elucidated due to limited information on its virulence factors. Here we report the identification of in vivo expressed antigens of E. faecalis by using a novel immunoscreening technique called change-mediated antigen technology (CMAT) and an experimental animal model of endodontic infection. Materials and Methods: Among 4,500 E. coli recombinant clones screened, 19 positive clones reacted reproducibly with hyperimmune sera obtained from rabbits immunized with E. faecalis cells isolated from an experimental endodontic infection. DNA sequences from 16 of these in vivo-induced (IVI) genes were determined. Results: Identified protein antigens of E. faecalis included enzymes involved in housekeeping functions, copper resistance protein, putative outer membrane proteins, and proteins of unknown function. Conclusions: In vivo expressed antigens of E. faecalis could be identified by using a novel immune-screening technique CMAT and an experimental animal model of endodontic infection. Detailed analysis of these IVI genes will lead to a better understanding of the molecular mechanisms involved in the endodontic infection of E. faecalis.

Utility of Structural Information to Predict Drug Clearance from in Vitro Data

  • Lee, So-Young;Kim, Dong-Sup
    • Interdisciplinary Bio Central
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    • v.2 no.2
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    • pp.3.1-3.4
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    • 2010
  • In the present research, we assessed the utility of the structural information of drugs for predicting human in vivo intrinsic clearance from in vitro intrinsic clearance data obtained by human hepatic microsome experiment. To compare with the observed intrinsic clearance, human intrinsic clearance values for 51 drugs were estimated by the classical methods using in vivo-in vitro scale-up and by the new methods using the in vitro experimental data and selected molecular descriptors of drugs by the forward selection technique together. The results showed that taking consideration of molecular descriptors into prediction from in vitro experimental data could improve the prediction accuracy. The in vitro experiment is very useful when the data can estimate in vivo data accurately since it can reduce the cost of drug development. Improvement of prediction accuracy in the present approach can enhance the utility of in vitro data.