• Title/Summary/Keyword: Ex vivo

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Gene Therapy Using GM-CSF Gene Transferred by a Defective Infectious Single-cycle Herpes Virus in Micro-residual Organotropic Head and Neck Squamous Cell Cancer Model (향장기성 두경부 편평세포암종의 미세잔존암 모델에서 GM-CSF 유전자를 이입시킨 제한복제성 헤르페스바이러스 벡터를 이용한 종양백신의 유전자 치료)

  • Kim Se-Heon;Choi Eun-Chang;Kim Han-Su;Chang Jung-Hyun;Kim Ji-Hoon;Kim Kwang-Moon
    • Korean Journal of Head & Neck Oncology
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    • v.19 no.1
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    • pp.25-33
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    • 2003
  • Background and Objectives: The Herpes Simplex type 2 Defective Infectious Single Cycle virus (DISC virus) is attenuated virus originally produced as viral vaccines but are also efficient gene transfer vehicle. The main goals of this study were to examine the efficiencies of the gene transfer using DISC vectors for various head and neck squamous cell carcinoma cell lines and to evaluate the efficacy of vaccination with DISC virus carrying a immunomodulatory genes (GM-CSF) as cancer therapy in a organotopic oral cavity squamous cell cancer model. Materials and Methods : We determinated the gene transfer efficiency of DISC virus by x-gal stain method and proved gene and protein expression of DISC-GMCSF transfected SCCVII cells by RT-PCR and ELISA method. Also we evaluated the ex vivo vaccination effects of SCCVII/GMCSF (DISC-GMCSF transfected SCCVII vaccine) vaccine on preventing the recurrence of micro-residual tumor. After the vaccination of SCCVII/GMCSF, specific cytotoxic T-cell responses was evaluated by CTL assay. Results: At an MOI of 10 DISC virus showed 64-88% of transfection rates in various head and neck squamous cancer cell lines. SCCVII cells transduced by DISC virus vector (MOI=10) carrying the GM-CSF gene, produced 4.5 nanogram quantities of GM-CSF per $10^6$ cells. In vivo vaccination using tumor cells transduced ex vivo with DISC-GMCSF resulted in better protection rate against subsequent tumor recurrence in organotopic oral cavity cancer model. Although tumor free survival rate was not statistically significantly increased in vaccination group (p=0.078), tumor specific cytotocic T-cell responses were significantly increased in SCCVII/GMCSF vaccination group. Conclusion: These data demonstrate that; 1) The DISC virus vector is capable of efficient gene transfer to various head and neck squamous cancer cell lines, 2) GM-CSF secreting genetically modified tumor vaccine (SCCVII/GMCSF) efficiently protected against tumor recurrence in organotopic micro-residual oral cavity cancer model and produced tumor specific cytotoxic T-cell response. DISC virus-mediated, cytokine gene transfer may prove to be useful as a clinical therapy for head and neck cancers.

Mitigation effects of red Platycodon grandiflorum extract on lipopolysaccharide-induced inflammation in splenocytes isolated from mice (홍도라지 추출물이 마우스에서 분리한 비장세포에서 lipopolysaccharide로 유도된 염증에 미치는 영향)

  • Park, Eun-Jung;Lee, You-Suk;Jeong, Hyun Cheol;Lee, Sung-Hyen;Lee, Hae-Jeung
    • Journal of Nutrition and Health
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    • v.52 no.3
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    • pp.243-249
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    • 2019
  • Purpose: Platycodon grandiflorum (PG) is known to have effective antimicrobial and anticancer activity. The main bioactive components of PG are saponins, and these could contribute to anti-inflammatory activity. However, little is known about the anti-inflammatory effect of PG. In this study, we aim to assess the anti-inflammatory response to Red PG Extract (RPGE) in splenocytes under ex vivo conditions. Methods: The cell viability of isolated splenocytes taken from mice was analyzed by performing a Cell Counting Kit-8 assay. The productions of nitric oxide (NO) and cytokines (specifically interleukin-6 (IL-6) and interleukin-10 (IL-10)) were measured utilizing Griess reagent and ELISA, respectively. Results: We found that co-treatment with RPGE and Lipopolysaccharide (LPS) decreased isolated splenocyte proliferation as compared with that of the LPS-stimulated control. We also observed that RPGE markedly suppressed NO synthesis and IL-6 production that was induced by LPS. There were no significant differences of IL-10 production between co-treatment with RPGE plus LPS and treatment with LPS alone. Conclusion: When taken together, our data has shown that RPGE mitigates LPS-induced inflammation in splenocytes isolated from mice. Further research is surely needed to confirm the anti-inflammation effects of RPGE in an in vivo model.

Carbon Nanotubes Multi Electrodes Array to Image Capacitance for Label-free Discrimination of Lipid Region in Atherosclerosis ex vivo

  • Song, Jun-Ho;Lee, Seon-Mi;Han, Nal-Ae;Yu, Gyeong-Hwa
    • Proceedings of the Korean Vacuum Society Conference
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    • 2016.02a
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    • pp.372.1-372.1
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    • 2016
  • Recently, there are a lot of diseases all around the world. Out of them, Atherosclerosis (AS) is the most common cause of stroke, cardiovascular mortality, and myocardial infarction. The macrophage-derived foam cell, which is formed by oxidized low-density lipoprotein (oxLDL), is the crucial marker for AS. In this study, we report a label-free capacitance imaging technique with multi-electrode array (MEA). The lipid-rich aorta arch lesions, which are derived from an apolipoprotein-E receptor-deficient (apoE-/-) mouse, exhibit higher capacitance than the lipid-free aorta arch, allowing the capacitance imaging of lipid region in atherosclerosis. To improve the contacts between MEA and tissue, polypyrrole(PPy)-coated multi walled carbon nanotubes (MWNTs) multi electrode array (PPy-MWNTs-MEA) was fabricated. Compared to TiN-MEA, PPy-MWNTs-MEA yielded lower contact impedance and better capacitance images. In addition, we have also developed a flexible MEA using single walled carbon nanotubes on a PET substrate. The lipid region could be discriminated in the capacitance images of the lipid-rich aorta arch lesions measured using flexible MEA, demonstrating a feasibility of in vivo applications.

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The Role of the Immune System in the use of Probiotic Lactic Acid Bacteria in Preventing and Treating Allergic Diseases

  • Choi, Kyeong-Ok;Nguyen, Hoang-Hai;Kwak, Hae-Soo
    • Food Science of Animal Resources
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    • v.30 no.1
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    • pp.1-12
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    • 2010
  • The immune system is generally divided into the innate and the adopted immune systems, both protecting the body from pathogens. Recently, allergies, a disease associated with an imbalanced immune system, have increased rapidly in developed countries. Prevailing symptoms of allergic diseases are eczema, allergic rhinitis, asthma, inflammatory bowel disease, and food allergy. Probiotic bacteria, mainly consisting of lactic acid bacteria, are used in the prevention and treatment of allergic diseases. The function of them is to stimulate the intestinal immune cells and form a complex signal network to activate other immune cells. Beneficial health effects of probiotics are based on the hygiene hypothesis, which suggests that sanitary environment is important for health, but limited exposure to environmental factors increases allergic diseases. An immunoregulatory effect of probiotic bacteria is demonstrated by controlled trial, animal model, in vitro, in vivo and ex vivo designs. However, the immunoregulatory effect of probiotic bacteria is controversial because it depends on probiotic strains, a dose and a type of diseases. In this review, we discussed clinical evidences on immunoregulatory effects of probiotic bacteria.

Non-Invasive in vivo Loss Tangent Imaging: Thermal Sensitivity Estimation at the Larmor Frequency

  • Choi, Narae;Kim, Min-Oh;Shin, Jaewook;Lee, Joonsung;Kim, Dong-Hyun
    • Investigative Magnetic Resonance Imaging
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    • v.20 no.1
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    • pp.36-43
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    • 2016
  • Visualization of the tissue loss tangent property can provide distinct contrast and offer new information related to tissue electrical properties. A method for non-invasive imaging of the electrical loss tangent of tissue using magnetic resonance imaging (MRI) was demonstrated, and the effect of loss tangent was observed through simulations assuming a hyperthermia procedure. For measurement of tissue loss tangent, radiofrequency field maps ($B_1{^+}$ complex map) were acquired using a double-angle actual flip angle imaging MRI sequence. The conductivity and permittivity were estimated from the complex valued $B_1{^+}$ map using Helmholtz equations. Phantom and ex-vivo experiments were then performed. Electromagnetic simulations of hyperthermia were carried out for observation of temperature elevation with respect to loss tangent. Non-invasive imaging of tissue loss tangent via complex valued $B_1{^+}$ mapping using MRI was successfully conducted. Simulation results indicated that loss tangent is a dominant factor in temperature elevation in the high frequency range during hyperthermia. Knowledge of the tissue loss tangent value can be a useful marker for thermotherapy applications.

In vitro Nasal Cell Culture Systems for Drug Transport Studies

  • Cho, Hyun-Jong;Termsarasab, Ubonvan;Kim, Jung-Sun;Kim, Dae-Duk
    • Journal of Pharmaceutical Investigation
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    • v.40 no.6
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    • pp.321-332
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    • 2010
  • Growing interest in the nasal route as a drug delivery system calls for a reliable in vitro model which is crucial for efficiently evaluating drug transport through the nasal cells. Various in vitro cell culture systems has thus been developed to displace the ex vivo excised nasal tissue and in vivo animal models. Due to species difference, results from animal studies are not sufficient for estimating the drug absorption kinetics in humans. However, the difficulty in obtaining reliable human tissue source limits the use of primary culture of human nasal epithelial cells. This shortage of human nasal tissue has therefore prompted studies on the "passage" culture of nasal epithelial cells. A serially passaged primary human nasal epithelial cell monolayer system developed by the air-liquid interface (ALI) culture is known to promote the differentiation of cilia and mucin gene and maintain high TEER values. Recent studies on the in vitro nasal cell culture systems for drug transport studies are reviewed in this article.

Involvement of Heme Oxygenase-1 in Orexin-A-induced Angiogenesis in Vascular Endothelial Cells

  • Kim, Mi-Kyoung;Park, Hyun-Joo;Kim, Su-Ryun;Choi, Yoon Kyung;Bae, Soo-Kyung;Bae, Moon-Kyoung
    • The Korean Journal of Physiology and Pharmacology
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    • v.19 no.4
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    • pp.327-334
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    • 2015
  • The cytoprotective enzyme heme oxygenase-1 (HO-1) influences endothelial cell survival, proliferation, inflammatory response, and angiogenesis in response to various angiogenic stimuli. In this study, we investigate the involvement of HO-1 in the angiogenic activity of orexin-A. We showed that orexin-A stimulates expression and activity of HO-1 in human umbilical vein endothelial cells (HUVECs). Furthermore, we showed that inhibition of HO-1 by tin (Sn) protoporphryin-IX (SnPP) reduced orexin- A-induced angiogenesis in vivo and ex vivo. Orexin-A-stimulated endothelial tube formation and chemotactic activity were also blocked in SnPP-treated vascular endothelial cells. Orexin-A treatment increased the expression of nuclear factor erythroid-derived 2 related factor 2 (Nrf2), and antioxidant response element (ARE) luciferase activity, leading to induction of HO-1. Collectively, these findings indicate that HO-1 plays a role as an important mediator of orexin-A-induced angiogenesis, and provide new possibilities for therapeutic approaches in pathophysiological conditions associated with angiogenesis.

Bleeding Time Prolongation Effect of Methanol Extract of Viscum album var. coloratum

  • Yang, Hyun-Ok;Park, Shin-Young;Hong, Kyung-Hee;Kang, Lin-Woo;Choe, Kwang-Hoon;Kim, Young-Kyoon
    • Natural Product Sciences
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    • v.8 no.4
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    • pp.152-154
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    • 2002
  • The methanol extract of Viscum album var. coloratum, Korean mistletoe, showed potent prolongation effects on the bleeding time in rats in vivo, and whole blood clotting time and plasma recalcification time in rats ex vivo. The prolongation effect on the bleeding time of Korean mistletoe is comparable to that of Viscum album L., European mistletoe, 185.6% and 176.5%, respectively. However, the water extracts of the both plants did not show any prolongation effects. Platelet activating factor (PAF) receptor binding assay was carried out to elucidate the action mechanisms of the extracts, and both of the methanol extracts did not show any inhibitory activity. The $LD_{50}$ of the methanol extracts of both mistletoes are more than 2 g/kg. These results suggest that the mehtanol extract of Korean mistletoe might be a potential candidate to develop new drug to improve microcirculation.

Enhancement of immune activities of Dioscorea japonica Thunberg in in vivo and ex vivo models

  • Lim, Seokwon;Kim, Na-Hyung;Cho, Hi-Jae;Jeong, Hyun-Ja
    • Korean Journal of Food Science and Technology
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    • v.51 no.4
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    • pp.398-403
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    • 2019
  • Dioscorea japonica Thunberg (DJ) has been widely used as a healthy food in Korea for the enhancement of physical stamina. Hence, the present study evaluated the immune-enhancing effect of DJ in forced swim test of mouse model. The immobility time of the group treated with DJ for 7 days was significantly reduced in comparison with that of the control group. After a forced swimming test, the changes in blood biochemical parameters and splenic T lymphocyte populations induced by the administration of DJ were assessed. Serum levels of lactic dehydrogenase, creatine phosphokinase, and aspartate aminotransferase were significantly decreased in DJ-administered group compared to the control group. However, administration of DJ did not affect the splenic T lymphocyte populations. Moreover, DJ significantly increased the production of interferon-g and interleukin-2 compared to the media control in splenocytes. Collectively, it may be concluded that DJ is useful for enhancement of physical and immune function.

Cochlear Implant Failure in the Pediatric Population

  • Ozer, Fulya;Yavuz, Haluk;Yilmaz, Ismail;Ozluoglu, Levent N.
    • Journal of Audiology & Otology
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    • v.25 no.4
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    • pp.217-223
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    • 2021
  • Background and Objectives: In cochlear implant (CI) surgery, the results and causes of revision and reimplantation may guide surgeons in establishing surgical protocols for revision surgery with safe audiological outcomes. The aim of this study was to review our experience in terms of etiology, surgical strategy, and hearing outcomes in pediatric patients who underwent CI removal and reimplantation. Subjects and Methods: All patients received implants of the same brand. Pre and postoperative Categories of Auditory Performance score and aided free-field pure tone audiometry thresholds were noted. In vivo integrity tests were performed for each patient and the results of ex vivo tests of each implant were obtained from manufacturer. Results: A total of 149 CIs were placed in 121 patients aged <18 years. The revision rate in children was 6.7% (10/121 children). Six patients had a history of head injury leading to a hard failure. The causes of reimplantation in others were soft failure (n=1), electrode migration (n=1), infection (n=1), and other (n=1). All patients showed better or similar postreimplantation audiological performance compared with pre-reimplantation results. Conclusions: It is very important to provide a safe school and home environment and educate the family for reducing reimplantation due to trauma. Especially for active children, psychiatric consultation should be continued postoperatively.