• Title/Summary/Keyword: Ex vivo

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M. tuberculosis Somatic Antigen Specific CD8+T cell Responses in BCG-Vaccinated Subjects (BCG 예방접종을 받은 개체에서 유도되어 있는 결핵균 균체항원에 특정한 CD8+T 세포의 보호 면역반응)

  • Cho, Jang-Eun;Cho, Sang-Nae;Lee, Kyung Wha;Park, Seung Kyu;Cho, Sungae
    • Tuberculosis and Respiratory Diseases
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    • v.59 no.3
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    • pp.272-278
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    • 2005
  • Background : The immune responses mediated by CD8+T cells are known to be significant in controlling M. tuberculosis infections. In order to determine the role of cytotoxic CD8+T cells in the protective immune mechanism in latently infected subjects, this study examined whether or not the cytotoxic immune responses of CD8+T cells specific to the M. tuberculosis somatic antigens are induced in BCG vaccinated healthy subjects. Methods : Cytotoxicity and $IFN-{\gamma}$ elispot assays were used to investigate the activities of CD8+T cells specific for the $thyA_{30-38}$ peptide epitope in circulating peripheral blood mononuclear cells (PBMC) from BCG-vaccinated HLA-A*0201 and A*0206 subjects. Results : The results indicate the cytotoxic and $IFN-{\gamma}$ immune responses of CD8+T cells specific for $thyA_{30-38}$ were induced in BCG vaccinated healthy subjects. Conclusion : The cytotoxic and $IFN-{\gamma}$ responses by CD8+T cells specific for the M. tuberculosis somatic antigens are induced in BCG-vaccinated subjects, and appear to be involved in the protective immune mechanism in latently infected people against a M. tuberculosis infection.

Effect of reperfusion with leukocyte-depleted blood on the expression of myocardial vascular cell adhesion molecule-1 (VCAM-1) and myocardial function in isolated working heart perfusion model (단순 작업성 심관류 모델에서의 신생돈 심장의 보존 후 백혈구-제거 혈액을 이용한 재관류가 심근 VCAM-1 발현 및 심기능에 미치는 영향)

  • 이정렬;석철준;서정욱;한재진
    • Journal of Chest Surgery
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    • v.33 no.3
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    • pp.213-220
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    • 2000
  • Background: Adhesion of leukocytes to myocardium or vascular endothelium has been known as an importation initial step in the ischemia-reperfusion injury which may affect the cardiac function. Therefore, leukocyte-depleted reperfusion may inhibit ischemia-reperfusion induced functional and ultrastructural deterioration. In this study, we quantified the time-dependent expression of the vascular cell adhesion molecule-1(VCMA-1) on piglet myocardium and demonstrated its relation to functional recovery using isolated piglet heart perfusion model. Material and Method: Neonatal(1 to 3 day old) piglet heart was harvested with 4$^{\circ}C$ University of Wisconsin solution (UWS) and presrved in the same solution for 12 hours. Ex vivo model of an isolated working neonatal piglet heart perfusion consisting of membrane oxygenator and roller-pump was used (Fig. 1). Hearts were grouped into leukocyte-non-depleted (group A, n=8) and leukocyte-depleted group(group B, n=8). In group B, hearts were reperfused with leukocyte-depleted blood using a leukocyte filter (Sepacell R, Asahi Medical, Japan). Segments of right atrium were taken before and after 1, 2, 3, and 4 hours of reperfusion for the evaluation of expression of VCAM-1. The intensity of immunohistochyemical satining of the VCAM-1 on the myocardium were graded semiquantitatively (0 to 4). For the evaluation of myocardial stroke work indices were calculated as well at the same time-points. Result: Mean expressins of VCAM-1 on the myocardium at 0, 1, 2, 3, adn 4 hours of reperfusion were 0.63, 1.44, 1.64, 2.65, and 3.34 in group A, while 0.56, 1.40, 1.50, 1.88 and 2.14 in group B (Fig. 3). Mean stroke work indices at 0.5, 1, 2, 3, and 4 hours after reperfusion were 1.35$\times$104, 1.32$\times$104, 1.14$\times$104, 0.81$\times$104, 0.68$\times$104 erg/gm in group A, while 1.40$\times$104, 1.43$\times$104, 1.43$\times$104, 1.28$\times$104, and 1.12$\times$104 erg/em in group B(Fig. 4). Conclusion : In this study, we demonstrated that leukocyte-depletion attenuated the expression of VCAM-1 during reperfusion and the time-dependent functional deterioration of the myocardium was well correlated with the degree of VCAM-1 expression.

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Three-dimensional Imaging with an Endoscopic Optical Coherence Tomography System for Detection of Airway Stenosis (기도협착 측정을 위한 내시경 광 결맞음 단층촬영법을 이용한 3차원 이미징)

  • Kwon, Daa young;Oak, Chulho;Ahn, Yeh-Chan
    • Korean Journal of Optics and Photonics
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    • v.30 no.6
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    • pp.243-248
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    • 2019
  • The respiratory tract is an essential part of the respiratory system involved in the process of respiration. However, if stenosis occurs, it interferes with breathing and can even lead to death. Asthma is a typical example of a reversible cause of airway narrowing, and the number of patients suffering from acute exacerbation is steadily increasing. Therefore, it is important to detect airway narrowing early and prevent the patient's condition from worsening. Optical coherence tomography (OCT), which has high resolution, is suitable for observing the microstructure of tissues. In this study we developed an endoscopic OCT system. We combined a 1300-nm OCT system with a servo motor, which can rotate at a high speed. A catheter was pulled back using a linear stage while imaging with 360° rotation by the motor. The motor was selected considering various requirements, such as torque, rotational speed, and gear ratio of pulleys. An ex vivo rabbit tracheal model was used as a sample, and the sample and catheter were immobilized by acrylic structures. The OCT images provided information about the structures of the mucosa and submucosa. The difference between normal and stenosed parts in the trachea was confirmed by OCT. Furthermore, through a three-dimensional (3-D) reconstruction process, it was possible to identify and diagnose the stenosis in the 3-D image of the airway, as well as the cross-sectional image. This study would be useful not only for diagnosing airway stenosis, but also for realizing 3-D imaging.

Transfection of Mesenchymal Stem Cells with the FGF-2 Gene Improves Their Survival Under Hypoxic Conditions

  • Song, Heesang;Kwon, Kihwan;Lim, Soyeon;Kang, Seok-Min;Ko, Young-Guk;Xu, ZhengZhe;Chung, Ji Hyung;Kim, Byung-Soo;Lee, Hakbae;Joung, Boyoung;Park, Sungha;Choi, Donghoon;Jang, Yangsoo;Chung, Nam-Sik;Yoo, Kyung-Jong;Hwang, Ki-Chul
    • Molecules and Cells
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    • v.19 no.3
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    • pp.402-407
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    • 2005
  • Bone marrow mesenchymal stem cells (MSCs) have shown potential for cardiac repair following myocardial injury, but this approach is limited by their poor viability after transplantation. To reduce cell loss after transplantation, we introduced the fibroblast growth factor-2 (FGF-2) gene ex vivo before transplantation. The isolated MSCs produced colonies with a fibroblast-like morphology in 2 weeks; over 95% expressed CD71, and 28% expressed the cardiomyocyte-specific transcription factor, Nkx2.5, as well as ${\alpha}$-skeletal actin, Nkx2.5, and GATA4. In hypoxic culture, the FGF-2-transfected MSCs (FGF-2-MSCs) secreted increased levels of FGF-2 and displayed a threefold increase in viability, as well as increased expression of the anti-apoptotic gene, Bcl2, and reduced DNA laddering. They had functional adrenergic receptors, like cardiomyocytes, and exposure to norepinephrine led to phosphorylation of ERK1/2. Viable cells persisted 4 weeks after implantation of $5.0{\times}10^5$ FGF-2-MSCs into infarcted myocardia. Expression of cardiac troponin T (CTn T) and a voltage-gated $Ca^{2+}$ channel (CaV2.1) increased, and new blood vessels formed. These data suggest that genetic modification of MSCs before transplantation could be useful for treating myocardial infarction and end-stage cardiac failure.

Effects of a Plant Water Extract Mixture(Ixeris sonchifolia Hance, Oenanthe javanica, Fagopyrum esculentum Moench, Hizikia fusiforme, Zingiber officinale Roscoe) on Mouse Immune Cell Activation (식물 혼합(고들빼기, 돌미나리, 메밀, 톳, 생강) 추출물이 마우스 면역 세포 활성에 미치는 영향)

  • Kim, Jung-Hee;Kim, Hyun-Sook;Ryu, Hye-Sook
    • The Korean Journal of Food And Nutrition
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    • v.20 no.1
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    • pp.74-78
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    • 2007
  • Ixeris sonchifolia Hance(Godulbaegi), Oenanthe javanica(Dolminari), Fagopyrum esculentum Moench(Buckwheat), Hizikia fusiforme(Seaweed Fusiforme) and Zingiber officinale Roscoe(Ginger) have all been used as one of the traditional remedies as well as food source. There are few studies However, on their immunomodulating effects have been reported. We previously reported that ex vivo supplementation of each of the Ish, Oj, Fem, Hf and Zor water extracts enhanced the splenocytes proliferation compared to the control group. In this study, the combined immunomodulative effects of a plant water extract mixture containing these five food sources(Ish+Oj+Fem+Hf+Zor) was compared to the individual effect of each. The production of cytokine(IL-1${\beta}$, IL-6, and TNF-${\alpha}$), secreted by macrophages stimulated with LPS or without, were detected via ELISA assay using a cytokine kit. After 48hrs of incubation with mitogen(ConA or LPS) stimulation, the mouse splenocyte proliferation in the experimental group had significantly increased at two different concentrations compared to the control group. The results of this study may suggest that the supplementing with a plant water extract mixture could regulate immune function by increasing splenocyte proliferation as well as enhance immune function by regulating the cytokine production capacity activated macrophages in mice.

Formation of Provitamin-$B_5$ Liquid Crystal with Hydrogenated Lecithin and Its Effectiveness of Moisturizing Activity (HL에 의한 프로비타민-$B_5$ 액정의 형성과 보습효과)

  • Kim, In-Young;Zhoh, Choon-Koo;Ryoo, Hee-Chang
    • Journal of the Korean Applied Science and Technology
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    • v.20 no.2
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    • pp.101-109
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    • 2003
  • Provitamin $B_5$ liquid crystal ($PVB_5$-LC) was the new emulsion system to enhance moisturizing activity on the skin. In this study, it should be mentioned that PVB5-LC could be prepared with the main compound of hydrogenated lecithin (HL) in oil-in water (O/W) emulsion. The key ingredient of humectants was contained 2% of provitamin $B_5$ ($PVB_5$) into the $PVB_5$-LC. The best suitable compositions of $PVB_5$-LC were made from 4.0 wt% of HL, 4.0 wt% of cetostearyl alcohol (CSA) as emulsifier and gelling agent, 3.0 wt% of 1,3-butylene glycol (1,3-BG) and 2.0 wt% of glycerin as moisturizers, 3.0 wt% of cyclomethicone (CMC), 3.0 wt% of isononanoate (ININ), 3.0 wt% of capric/caprylic triglyceride (CCTG), 3.0 wt% of macadamia nut oil (MNO) as emollients. As the analytical result of $PVB_5$-LC, it could know that the distribution range of particle size was 0.14 to 12.37 m level (mean size 3.24 m). It was certified the multi lamellar phase around the droplet of liquid crystal when observed the droplet particles through a polarization microscope. And it clinically was tested the effectiveness of moisturizing activity (in-vivo) compared with control sample (O/W emulsion). The effectiveness of moisturizing activity of $PVB_5$-LC with Skincon-200EX after 6 hours went up 49.0% (p<0.05, n=20) whereas the effectiveness of moisturizing activity of generally O/W emulsion cream was 25.7% (p<005, n=20). Also, in case of Comeometer CM-825, the moisturizing activity of $PVB_5$-LC after 6 hours rose 36.6% (p<0.05, n=20) whereas the effectiveness of moisturizing activity of generally O/W emulsion cream was 10.8%. Therefore, it was known that the effectiveness moisturizing activity of $PVB_5$-LC with HL was remarkably superior compared with O/W emulsion cream.

Biological Activities of Flavonoid Glycosides Isolated from Angelica keiskei (신선초에서 분리된 flavonoid glycosides의 생리활성)

  • Shim, Jae-Seok;Kim, Seung-Deok;Kim, Tae-Seok;Kim, Kyung-Nam
    • Korean Journal of Food Science and Technology
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    • v.37 no.1
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    • pp.78-83
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    • 2005
  • Recently, much attention has been focused on plant antioxidants, because they are expected to protect against oxidative damage, possibly preserving biological functions of cells. Antioxidant compounds were isolated from Angelica keiskei through extraction with 80% EtOH, and fractionations were carried out sequentially with n-hexane, chloroform, ethyl acetate, n-butanol, and water. Two active compounds were isolated from ethyl acetate fraction by silica gel column chromatography, and were identified as isoquercitrin ($quercetin-3-O-{\beta}-D-glucose$) and hyperoside ($quercetin-3-O-{\beta}-D-glucose$). Isoquercitrin and hyperoside showed strong antioxidative potency, as revealed by evaluation of their ABTS, DPPH, OH, and $H_{2}O_{2}$ radical-scavenging activities, and ex vivo DNA damage-protecting effects.

Activation of Intestinal Immune System by an Orally Administered Methanol Extract from Pine Needles (솔잎 메탄올추출물의 마우스 경구투여에 의한 장관면역 활성)

  • Yoon, Jin-A;Yu, Kwang-Won;Shin, Shun-Hyuck;Cho, Hong-Yeon
    • Journal of the Korean Society of Food Science and Nutrition
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    • v.39 no.3
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    • pp.356-362
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    • 2010
  • When pine (Pinus densiflora) needles were fractionated into cold water (PD-CW), hot water (PD-HW) and methanol extract (PD-M), PD-M showed potent simulating activity (1.19-fold of the saline control) for proliferation of bone marrow cells mediated by Peyer's patch cells in vitro. MeOH extracts were prepared by homogenization, stirring or reflux to identify the method of methanol extraction, and MeOH extract by reflux method showed significantly highest intestinal immune system modulating activity (1.30-fold) in vitro. The intestinal immune system modulating effect of orally administered PD-M fractionated from pine needles also were studied in mice. Analyzing intestinal immune system modulating activity mediated Peyer's patch cells from C3H/He mice which had been fed with PD-M at different doses for 7 days, 1.0 g/kg of BW/day indicated that the bone marrow cells had proliferated (3.65-fold of 3% EtOH administered group). In addition, the amounts of IL-6 in the culture supernatant of Peyer's patch cells at 1.0 g/kg of BW/day were increased (1.13-fold) whereas the production of GM-CSF was not dose dependent. These results indicate that oral administration of PD-M enhances the secretion of hematopoietic growth factors such as GM-CSF and IL-6 from Peyer's patch cells, and these cytokines also act on modulator of bone marrow cell proliferation.

Ex vivo High-resolution Optical Coherence Tomography (OCT) Imaging of Pleural Reaction after Pleurodesis Using Talc

  • Ahn, Yeh-Chan;Oak, Chulho;Park, Jung-Eun;Jung, Min-Jung;Kim, Jae-Hun;Lee, Hae-Young;Kim, Sung Won;Park, Eun-Kee;Jung, Maan Hong
    • Journal of the Optical Society of Korea
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    • v.20 no.5
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    • pp.607-613
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    • 2016
  • The pleura is known as an end target organ of exposure to toxic environmental materials such as fine particulate matter and asbestos. Moreover, long-term exposure to hazardous materials can eventually lead to fatal lung disease such as diffuse pleural fibrosis or mesothelioma. Chest computed tomography (CT) and ultrasound are gold standard imaging modalities for detection of advanced pleural disease. However, a diagnostic tool for early detection of pleural reaction has not been developed yet due to difficulties in imaging ultra-fine structure of the pleura. Optical coherence tomography (OCT), which provides cross-sectional images of micro tissue structures at a resolution of 2-10 μm, can image the mesothelium with a thickness of ~100 μm and therefore enables investigation of the early pleural reaction. In this study, we induced the early pleural reaction according to a time sequence after pleurodesis using talc, which has been widely used in the clinical field. The pleural reaction in talc grouped according to the time sequence (1st, 2nd, 4th weeks) showed a significant thickening (average thickness: 45 ± 7.5 μm, 80 ± 10.7 μm, 90 ± 12.5 μm), while the pleural reaction in sham and normal groups showed pleural change from normal to minimal thickening (average thickness: 16 ± 5.5 μm, 17 ± 4.5 μm, 15 ± 6.5 μm, and 12 ± 7.5 μm, 13 ± 2.5 μm, 12 ± 3.5 μm). The measurement of pleural reaction by pathologic examinations was well-matched with the measurement by OCT images. This is the first study for measuring the thickness of pleural reactions using a biophotonic modality such as OCT. Our results showed that OCT can be useful for evaluating the early pleural reaction.

Effects of Haedoksamul-tang on Trimellitic Anhydride-induced Contact Hypersensitivity in a Mouse Model (해독사물탕(解毒四物湯)이 피부 접촉성 과민반응에 미치는 영향(影響))

  • Jeon, Young-Kyun;Jung, Myung;Bok, Young-Ok;Lee, Eun-Hye;Lim, Kyu-Sang;Yun, Young-Gab
    • The Journal of Korean Medicine Ophthalmology and Otolaryngology and Dermatology
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    • v.29 no.2
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    • pp.65-81
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    • 2016
  • Objectives : Haedoksamul-tang (HSTE), a water extract from a mixture of Phellodendri Cortex, Coptidis, Scutellariae Radix, Gardeniae Fructus, Angelica acutiloba Radix, Cnidii Rhizoma, Paeoniae Radix, Rehmanniae Radix, has been traditionally used for allergic skin diseases such as atopic dermatitis and contact dermatitis in oriental countries. However, little is known about the effects of aqueous extract of HSTE on trimellitic anhydride (TMA)-induced contact hypersensitivity (CHS) in a mouse model. Methods : In this study, we investigate the pharmacological effects of HSTE on TMA-induced CHS in Balb/c mice. Contact hypersensitivity was induced in mice by topically sensitizing and challenging with TMA in flank skin and ears during oral administration (for 17 days) and topical treatment (30 min before challenge) with HSTE. We examined the effects of HSTE on IgE and IgG1 levels, inflammatory parameters in ear tissues, CD4+/CD8+ ratio, cytokine and chemokine production in sera, tissues, and immune cells from TMA-sensitized mice.Results : Oral and topical administration with HSTE reduced, in a dose dependent manner, thickness and leukocyte infiltration of ear tissues and IgE levels in serum from mice sensitized with TMA. In addition, auricula lymph node cells isolated from TMA-sensitized mice significantly elevated the expression ratio of CD4+/CD8+ as well as increased the production of IL-4, IL-5, IL-13 and IFN-γ by ex vivo stimulation with antibodies against CD3 and CD28, and these inflammatory indexes, except for IFN-γ, were significantly suppressed by orally and topically administration of HSTE. Furthermore, stimulation of auricula lymph node cells from TMA-sensitized mice with antibodies against CD3 and CD28 increased the production of MCP-1/CCL2 and MIP-1α/CCL3, and these effects were inhibited in a dose-dependent manner in cells from mice treated with HSTE. Conclusions : These results suggest that HSTE can be used for treating contact hypersensitivity by inhibiting leukocyte infiltration as well as production of serum IgE and chemokine/Th2 cytokine in an animal model.