• Title/Summary/Keyword: ex vivo study

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A Study of the Generation of Transgenic Chickens That Express Human SOD-3 Protein (사람의 SOD-3 단백질을 발현하는 형질전환 닭 생산 연구)

  • Byun, S.J.;Park, C.;Kim, J.A.;Woo, J.S.;Lee, H.C.;Kim, T.Y.;Kim, S.H.;Seong, H.H.;Park, J.K.;Jeon, I.S.
    • Korean Journal of Poultry Science
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    • v.35 no.3
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    • pp.241-245
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    • 2008
  • Lentiviral vector system is efficient vehicles for the delivery of exogenous genes, and it is generally used in the generation of transgenic chickens. In this study, we used recombinant lentiviral vectors to generate transgenic chicks that express the human superoxide dismutase-3 gene driven by the chicken ovalbumin promoter. It is well known that superoxide dismutases(SODs) are believed to play a crucial role in protecting cells against oxygen toxicity. There are three forms of SOD proteins: cytosolic Cu-Zn SOD, mitochondrial Mn SOD, and extracellular SOD(SOD-3). The recombinant lentivirus containing the human SOD-3 gene was injected into the subgerminal cavity of freshly laid eggs. Subsequently, the embryos were incubated to hatch using phases II and III of the surrogate shell ex vivo culture system. From 341 injected embryos, the 78 chicks hatched after 21 days incubation. The hatched chicks were screened for the human SOD-3 gene by using PCR. Two of 47 male chickens that survived to sexual maturity contained the human SOD-3 gene in their semen. These results showed that our transgenic chicken generation system was completely established.

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.

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.

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.

Evaluation of the Femoral Stem Implant in Canine Total Hip Arthroplasty: A Cadaver Study

  • Cho, Hyoung Sun;Kwon, Yonghwan;Kim, Young-Ung;Kang, Jin-Su;Lee, Kichang;Kim, Namsoo;Kim, Min Su
    • Journal of Veterinary Clinics
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    • v.36 no.1
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    • pp.53-61
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    • 2019
  • Total hip arthroplasty (THA) is a successful surgical treatment for both patients with chronical lameness and dogs who are nonresponsive to medical treatments, providing excellent joint function for returning dogs to the normal gait in 80% to 98% of hip dysplasia (HD) patients. The THA surgical implant system manufactured by BioMedtrix and Kyon are today widely accepted. When comparing the BioMedtrix biological fixation (BFX) system to the BioMedtrix cemented fixation (CFX) system, the many advantages of BFX, which include longer potential implant life, decreased risk of postoperative or later infection, and better implant stability, become evident. However, BFX implies a greater risk of femoral fracture during reaming and requires a more precise surgical technique to achieve good implant fit, given the press-fit nature of cementless THA. The purposes of this study are to both describe the mistakes and complications during stem implantation for beginner surgeons with both the BFX and the CFX systems and to document the initial result of 12 implantations in canine cadavers. Given the detailed evaluations of 3 specialists, who are Diplomate American College of Veterinary Surgeons (DACVS), only 3 of 11 stems were appropriately sized. Specifically, 6 stems were anteverted rather than being retroverted; further, although 7 stems were coaxial with the femoral long axis in the frontal plane, the other stems were in the varus at the frontal plane, with the proximal medial stem adjacent to the medial femoral cortex. Moderate angulation from the cranial to the caudal directions was found in 4 cases in the sagittal plane. Additionally, 1 case of femoral fissure and 1 case of perforated femoral cortex were reported. It is not easy for surgeons performing cementless THA for the first time to achieve a good result, even though they completed an educational course about it and given that catastrophic complications often occurred during early surgical clinical cases. Therefore, ex-vivo studies are sincerely required to get an expertise by rehearsing the preparation of the femoral envelop in isolated bones. Further studies should be conducted to achieve both highly accurate implant size and correct orientation during the preoperative planning. Additionally, surgeons' learning curve should be examined in future investigations.

Evaluation of Therapeutic Monitoring of Prostate Cancer (PCa) using [18F]Florastamin, Diagnostic Radiopharmaceutical for PCa: Non-clinical Ex vivo Whole-body Autoradiographic Analysis

  • Min Hwan Kim;Kyongkyu Lee;Hee Seup Kil;Soon Jeong Kwon;Yong Jin Lee;Kyo Chul Lee;Dae Yoon Chi
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.9 no.1
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    • pp.17-21
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    • 2023
  • In this study, we evaluated the targeting of prostate cancer (PCa) using [18F]Florastamin in non-clinical study, for the purpose of therapeutic monitoring of [177Lu]Ludotadipep, a therapeutic radiopharmaceutical for PCa, [18F]Florastamin/[177Lu]Ludotadipep was co-administered to a single-individual prostate tumor bearing mouse model, mimicking clinical condition. Considering the difference in half-life of the two isotopes (18F or 177Lu), image scan of whole-body autoradiography was performed at 24 or 48 h after preparation of frozen section, respectively. Then, it was confirmed whether they showed the same targeting efficiency for the area of tumor. A tumor xenograft model was prepared using PSMA-overexpressing PC3-PIP prostate cancer cells. [18F]Florastamin [111 MBq (3 mCi) in 100 µL]/177Lu]Ludotadipep [3.7 MBq (100 µCi) in 100 µL] was co-administered through the tail vein, and 2 hours after administration, the mice were frozen, and after freezing for 24 hours, whole-body cryosection was performed at 24 h after freezing. Image scanning using cryosection was performed after 24 or 48 hours after freezing, respectively. In the scan image after 24 hours, tumor uptake of [18F] Florastamin/[177Lu]Ludotadipep were simultaneously observed specific uptake in the tumor. In the scan image after 48 hours in the same section, signal of 18F was lost by decay of radioisotope, and specific uptake image for [177Lu]Ludotadipep was observed in the tumor. Uptake of [177Lu]Ludotadipep was specific to the same tumor region where [18F]Florastamin/[177Lu]Ludotadipep was uptake. These results suggested that [18F]Florastamin showed the same tumor uptake efficiency to PCa as [177Lu]Ludotadipep, and effective therapeutic monitoring is expected to be enable using [18F]Florastamin during [177Lu]Ludotadipep therapy for PCa.

Dysregulation of Cellular Immune Functions on Gastric Administration to Mixtures of Polyethlene Microplastics and Metallic Lead in Mice (폴리에틸렌 미세플라스틱과 납의 복합노출에 따른 실험동물의 세포면역기능 조절 장애 평가)

  • Gyoungwoo Lee;Changyul Kim
    • Journal of Environmental Health Sciences
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    • v.49 no.1
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    • pp.22-29
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    • 2023
  • Background: The existing research results on the combined toxicity of these pollutants using mammals, such as rodents, are insufficient, especially in relation to changes in the immune system. Objectives: This study aims at evaluating the cellular immune response to PE-MPs solely or when combined with Pb, which possess excellent adsorption capacity with PE-MPs and is commonly co-exposed in our daily lives. Methods: The study investigated the cellular immune function of 9-week ICR mice with 28 days exposure to PE-MPs (2 mg/mouse/day) and Pb (0.1 mM in distilled water) individually and in combination. PE-MPs were administered via gastric intubation while the lead intake was conducted via the oral drinking water route. Cellular immunity was evaluated by analyzing the production for TH1 cytokines namely, TNF-α and IFN-𝛾 and TH2 cytokines, IL-4 and IL-6 in culture supernatants from polyclonally activated splenic mononuclear cells ex vivo. Results: Both the PE-MPs only and the PE-MPs+Pb exposure group revealed an increased TH1 response with elevated TNF-α and IFN-𝛾 levels and downregulated TH2 response with low IL-4, and IL-6 production levels compared to the control group. Furthermore, an increased IFN-𝛾/IL-4 ratio was found in the PE-MPs only and PE-MPs+Pb exposure groups, which indicated the skewedness to TH1 response. Meanwhile, reduced blood hemoglobin levels and increased levels of IL-4, the dominant TH2 cytokine in the Pb-only exposure group, were observed. Conclusions: Our current findings on the predominance of TH1 immune response in the PE-MPs and PE-MPs+Pb groups suggest that PE-MPs could be responsible for the predominant induction of the cellular immune changes. This finding could be used as an important landmark in research related to TH1 predominance, such as autoimmune diseases. It suggests that additional research on immune modulation using longer exposure durations or the same exposure route is required to elucidate stronger findings.

Development of Phantom and Comparison Analysis for Performance Characteristics of MOSFET Dosimeter (MOSFET 선량계 특성분석을 위한 팬톰 개발 및 특성 비교)

  • Chung, Jin-Beom;Lee, Jeong-Woo;Kim, Yon-Lae;Lee, Doo-Hyun;Choi, Kyoung-Sik;Kim, Jae-Sung;Kim, In-Ah;Hong, Se-Mie;Suh, Tae-Suk
    • Progress in Medical Physics
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    • v.18 no.1
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    • pp.48-54
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    • 2007
  • This study is to develope a phantom for MOSFET (Metal Oxide Semiconductors Field Effect Transistors) dosimetry and compare the dosimetric properties of standard MOSFET and microMOSFET with the phantom. In this study, the developed phantom have two shape: one is the shape of semi-sphere with 10cm diameters and the other one is the flat slab of $30{\times}30cm$with 1 cm thickness. The slab phantom was used for calibration and characterization measurements of reproducibility, linearity and dose rate dependency. The semi-sphere phantom was used for angular and directional dependence on the types of MOSFETs. The measurements were conducted under $10{\times}10cm^2$ fields at 100cm SSD with 6MV photon of Clinac (21EX, Varian, USA). For calibration and reproducibility, five standard MOSFETS and microMOSFETs were repeatedly Irradiated by 200cGy five times. The average calibration factor was a range of $1.09{\pm}0.01{\sim}1.12{\pm}0.02mV/cGy$ for standard MOSFETS and $2.81{\pm}0.03{\sim}2.85{\pm}0.04 mV/cGy$ for microMOSFETs. The response of reproducibility in the two types of MOSFETS was found to be maximum 2% variation. Dose linearity was evaluated In the range of 5 to 600 cGy and showed good linear response with $R^2$ value of 0.997 and 0.999. The dose rate dependence of standard MOSFET and microMOSFET was within 1% for 200 cGy from 100 to 500MU/min. For linearity, reproducibility and calibration factor, two types of MOSFETS showed similar results. On the other hand, the standard MOSFET and microMOSFET were found to be remarkable difference in angular and directional dependence. The measured angular dependence of standard MOSFET and microMOSFET was also found to be the variation of 13%, 10% and standard deviation of ${\pm}4.4%,\;{\pm}2.1%$. The directional dependence was found to be the variation of 5%, 2% and standard deviation of ${\pm}2.1%,\;{\pm}1.5%$. Therefore, dose verification of radiation therapy used multidirectional X-ray beam treatments allows for better the use of microMOSFET which has a reduced angular and directional dependence than that of standard MOSFET.

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Tetrahydrobiopterin Inhibits PDGF-stimulated Migration and Proliferation in Rat Aortic Smooth Muscle Cells via the Nitric Oxide Synthase-independent Pathway

  • Jiang, Xiaowen;Kim, Bo-Kyung;Lin, Haiyue;Lee, Chang-Kwon;Kim, Jung-Hwan;Kang, Hyun;Lee, Pil-Young;Jung, Seung-Hyo;Lee, Hwan-Myung;Won, Kyung-Jong
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.3
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    • pp.177-183
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    • 2010
  • Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.