• Title/Summary/Keyword: ex vivo study

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Effect of MCT (medium-chain triglyceride) and LCT (long-chain triglyceride) on Myocardial Ischemia/Reperfusion Injury and Platelet Aggregation in Rat (MCT(medium-chain triglyceride) 및 LCT(long-chain triglyceride) 유제가 백서에서 허혈/재관류 심장기능손상 및 혈소판응집능에 미치는 영향)

  • Lee, Soo-Hwan;Jung, Yi-Sook;Hong, Jeong;Kim, Min-Hwa;Lee, Hee-Joo;Baik, Eun-Joo;Wang, Hee-Jung;Kim, Myung-Wook;Moon, Chang-Hyun
    • Biomolecules & Therapeutics
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    • v.6 no.4
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    • pp.358-363
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    • 1998
  • Intravenous lipid emulsion is used extensively as a major component of parenteral nutrition for patients in the surgical intensive care unit. Abnormal cardiovascular function related to lipid infusion has been reported although conflicting results exist. In the present study, we investigated the effects of intravenous emulsions of long-chain triglyceride (LCT) and medium-chain triglyceride (MCT) on myocardial ischemia/ reperfusion injury and on platelet aggregation in rat. There was no difference between LCT and MCT considering the effects on left ventricular developed pressure (LVDP) and coronary flow rate (CFR) before and after ischemia/reperfusion in isolated rat heart. On the other hand, a difference was found between LCT and MCT with regard to their effects on heart rate (HR) and end diastolic pressure (EDP) after ischemia/reperfusion. After ischemia/reperfusion, HR was significantly (P<0.05) reduced and EDP significantly (P<0.05) inc.eased by LCT (18$\pm$2.0% and 42.8$\pm$8.9%, respectively), but not by MCT Ex vivo platelet aggregation induced by collagen was reduced by LCT infusion, but not by MCT These findings suggest that MCT may have slightly more favorable effect than LCT on the myocardial function after ischemia/reperfusion in rat.

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Mycobacterium tuberculosis Derived Epitope Peptide Specific CD8+T Cell Responses in Tuberculous Pleurisy

  • Cho, Jang-Eun;Kim, Young-Sam;Park, Moo-Suk;Lee, Kyung-Wha;Lee, Eun-Hee;Cho, Sang-Nae;Cho, Sung-Ae
    • Biomedical Science Letters
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    • v.13 no.4
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    • pp.325-332
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    • 2007
  • Cell-mediated immune response (CMI) is a major immune protective mechanism against tuberculosis (TB) infection. Among several components involved in CMI, recent studies suggest that CD8+ T cells are important in controlling TB infection. In our previous report, we defined four Mycobacterium tuberculosis (MTB) derived epiotpe peptides specific for HLA-A*0201-restricted CD8+ T cells. These four peptides are $PstAl_{75-83}$, $ThyA_{30-38}$, $RpoB_{127-135}$ and $85B_{15-23}$. In this study, these epitope peptides specific CD8+ T cell responses in tuberculous pleurisy were investigated using ex vivo $IFN-\gamma$ elispot assay and intracellular $IFN-\gamma$ staining method. As a result, we observed these epitope peptide specific CD8+ T cell responses are induced in all three patients with tuberculous pleurisy suggesting that CD8+ T cells are involved in protective immune mechanism against MTB infection in tuberculous pleurisy. However, the CMI to mitogens and MTB antigens from pleural fluids of patients with tuberculous pleurisy does not seem to correlate with that from peripheral blood, although the sample size is too small to make any conclusion. In sum, the MHC I restricted CD8+ T cell responses seem to be induced efficiently in the pleural fluids, at the site of TB infection, in which the CMI is actively induced. In addition, these experiments suggest that MHC I restricted CD8+ T cell mediated immune responses are also involved in protective mechanism against MTB infection in extra-pulmonary TB.

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Human extracellular superoxide dismutase (EC-SOD) expression in transgenic chicken

  • Byun, Sung June;Ji, Mi-Ran;Jang, Ye-Jin;Hwang, A-In;Chung, Hee Kyoung;Kim, Jeom Sun;Kim, Kyung-Woon;Chung, Hak-Jae;Yang, Byoung-Chul;Jeon, Iksoo;Park, Jin-Ki;Yoo, Jae Gyu;Kim, Tae-Yoon
    • BMB Reports
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    • v.46 no.8
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    • pp.404-409
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    • 2013
  • Extracellular superoxide dismutase (EC-SOD) is a metallo-protein and functions as an antioxidant enzyme. In this study, we used lentiviral vectors to generate transgenic chickens that express the human EC-SOD gene. The recombinant lentiviruses were 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. Of 158 injected embryos, 16 chicks (G0) hatched and were screened for the hEC-SOD by PCR. Only 1 chick was identified as a transgenic bird containing the transgene in its germline. This founder (G0) bird was mated with wild-type hens to produce transgenic progeny, and 2 transgenic chicks (G1) were produced. In the generated transgenic hens (G2), the hEC-SOD protein was expressed in the egg white and showed antioxidant activity. These results highlight the potential of the chicken for production of biologically active proteins in egg white.

Poncirin alleviates the symptoms of dextran sulfate sodium-induced colitic mice (Poncirin의 dextran sulfate sodium 유도 마우스 궤양성 대장염 증세 감소 효과)

  • Kim, Jong-Bin;Cho, Woong;Han, Ar-Reum;Seo, Eun-kyung;Lee, Kyung-Tae
    • Korean Journal of Pharmacognosy
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    • v.39 no.2
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    • pp.104-109
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    • 2008
  • We previously reported that anti-inflammatory properties of poncirin, isolated from fruit of Poncirus trifoliata, might be the result from the inhibition of inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), tumor necrosis $factor-{\acute{a}}$ ($TNF-{\alpha}$) and interlukin-6 (IL-6) expression via the down-regulation of $NF{-\kappa}B$ binding activity. In this study, we investigated whether poncirin has an inhibitory effect on the production of pro-inflammatory mediators ex vivo and whether poncirin could relieve the symptoms of dextran sulfate sodium (DSS)-induced colitis in mice model of inflammatory bowel disease. Poncirin significantly inhibited the productions of NO, IL-6 and $TNF-{\alpha}$ in lipopolysaccharide (LPS)-induced mouse peritoneal macrophage. In addition, poncirin-treated mice when compared to control mice not receiving treatment recovered better from the weight loss caused by DSS-induced colitis. Changes in disease activity index (DAI) of poncirin-treated mice were also more favorable than for control mice and were comparable with mice treated with a typical anti-inflammatory-drug, 5-aminosalichylic acid (5-ASA). In addition, suppression of plasma NO and IL-6 productions of poncirin-treated mice was also observed in DSS-induced colitis. These results suggest that poncirin has potentially useful anti-inflammatory effects mediated by suppression of inflammatory mediator productions.

Biological Control of Apple Ring Rot on Fruit by Bacillus amyloliquefaciens 9001

  • Li, Yan;Han, Li-Rong;Zhang, Yuanyuan;Fu, Xuechi;Chen, Xinyi;Zhang, Lixia;Mei, Ruhong;Wang, Qi
    • The Plant Pathology Journal
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    • v.29 no.2
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    • pp.168-173
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    • 2013
  • Apple ring rot disease, caused by Botryosphaeria dothidea (Moug. ex. Fr) Ces. et de Not., is one of the most important diseases on apple fruits. In this study, strain 9001 isolated from healthy apple fruits from an infested orchard was evaluated for its biocontrol activity against apple ring rot in vitro and in vivo. Strain 9001 showed obvious antagonistic activity to B. dothidea YL-1 when plated on potato dextrose agar. Soaking healthy apples in the bacterial suspensions of strain 9001 prior to artificial inoculation of fungal pathogen resulted in a dramatic decrease in disease incidence when compared to the control. Moreover, either field application in the growth season or postharvest treatment of apples from infected orchards with bacterial suspensions of strain 9001 resulted in significantly reduced disease incidence within the storage period for 4 months at room temperature. Based on the phylogenetic analysis of 16S rRNA and the gyrA gene, strain 9001 was identified as Bacillus amyloliquefaciens. These results indicated that B. amyloliquefaciens 9001 could be a promising agent in biocontrol of apple ring rot on fruit, which might help to minimize the yield loss of apple fruit during the long postharvest period.

Development of an Optical Tissue Clearing Laser Probe System

  • Yeo, Changmin;Kang, Heesung;Bae, Yunjin;Park, Jihoon;Nelson, J. Stuart;Lee, Kyoung-Joung;Jung, Byungjo
    • Journal of the Optical Society of Korea
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    • v.17 no.4
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    • pp.289-295
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    • 2013
  • Although low-level laser therapy (LLLT) has been a valuable therapeutic technology in the clinic, its efficacy may be reduced in deep tissue layers due to strong light scattering which limits the photon density. In order to enhance the photon density in deep tissue layers, this study developed an optical tissue clearing (OTC) laser probe (OTCLP) system which can utilize four different OTC methods: 1) tissue temperature control from 40 to $10^{\circ}C$; 2) laser pulse frequency from 5 to 30 Hz; 3) glycerol injection at a local region; and 4) a combination of the aforementioned three methods. The efficacy of the OTC methods was evaluated and compared by investigating laser beam profiles in ex-vivo porcine skin samples. Results demonstrated that total (peak) intensity at full width at half maximum of laser beam profile when compared to control data was increased: 1) 1.21(1.39)-fold at $10^{\circ}C$; 2) 1.22 (1.49)-fold at a laser pulse frequency of 5 Hz; 3) 1.64 (2.41)-fold with 95% glycerol injection; 4) 1.86 (3.4)-fold with the combination method. In conclusion, the OTCLP system successfully improved the laser photon density in deep tissue layers and may be utilized as a useful tool in LLLT by increasing laser photon density.

Human umbilical cord blood plasma alleviates age-related olfactory dysfunction by attenuating peripheral TNF-α expression

  • Lee, Byung-Chul;Kang, Insung;Lee, Seung-Eun;Lee, Jin Young;Shin, Nari;Kim, Jae-Jun;Choi, Soon Won;Kang, Kyung-Sun
    • BMB Reports
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    • v.52 no.4
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    • pp.259-264
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    • 2019
  • Social requirements are needed for living in an aging society and individual longevity. Among them, improved health and medical cares, appropriate for an aging society are strongly demanded. Human cord blood-derived plasma (hUCP) has recently emerged for its unique anti-aging effects. In this study, we investigated brain rejuvenation, particularly olfactory function, that could be achieved by a systemic administration of young blood and its underlying mechanisms. Older than 24-month-old mice were used as an aged group and administered with intravenous injection of hUCP repetitively, eight times. Anti-aging effect of hUCP on olfactory function was evaluated by buried food finding test. To investigate the mode of action of hUCP, brain, serum and spleen of mice were collected for further ex vivo analyses. Systemic injection of hUCP improved aging-associated olfactory deficits, reducing time for finding food. In the brain, although an infiltration of activated microglia and its expression of cathepsin S remarkably decreased, significant changes of proinflammatory factors were not detected. Conversely, peripheral immune balance distinctly switched from predominance of Type 1 helper T (Th1) cells to alternative regulatory T cells (Tregs). These findings indicate that systemic administration of hUCP attenuates age-related neuroinflammation and subsequent olfactory dysfunction by modulating peripheral immune balance toward Treg cells, suggesting another therapeutic function and mechanism of hUCP administration.

Endoscopic Laser Irradiation Condition of Urethra in Tubular Structure (관형 요도 조직 대상 내시경적 레이저 조사 조건 연구)

  • Hwarang, Shin;Seonghee, Lim;Yeachan, Lee;Hyun Wook, Kang
    • Journal of Biomedical Engineering Research
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    • v.44 no.1
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    • pp.85-91
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    • 2023
  • Stress urinary incontinence (SUI) occurs when abdominal pressure increases, such as sneezing, exercising, and laughing. Surgical and non-surgical treatments are the common methods of SUI treatment; however, the conventional treatments still require continuous and invasive treatment. Laser have been used to treat SUI, but excessive temperature increase often causes thermal burn on urethra tissue. Therefore, the optimal conditions must be considered to minimize the thermal damage for the laser treatment. The current study investigated the feasibility of the laser irradiation condition for SUI treatment using non-ablative 980 nm laser from a safety perspective through numerical simulations. COMSOL Multiphysics was used to analyze the numerical simulation model. The Pennes bioheat equation with the Beer's law was used to confirm spatio-temporal temperature distributions, and Arrhenius equation defined the thermal damage caused by the laser-induced heat. Ex vivo porcine urethral tissue was tested to validate the extent of both temperature distribution and thermal damage. The temperature distribution was symmetrical and uniformly observed in the urethra tissue. A muscle layer had a higher temperature (28.3 ℃) than mucosal (23.4 ℃) and submucosal layers (25.5 ℃). MT staining revealed no heat-induced collagen and muscle damage. Both control and treated groups showed the equivalent thickness and area of the urethral mucosal layer. Therefore, the proposed numerical simulation can predict the appropriate irradiation condition (20 W for 15 s) for the SUI treatment with minimal temperature-induced tissue.

Development and Degeneration of Retinal Ganglion Cell Axons in Xenopus tropicalis

  • Choi, Boyoon;Kim, Hyeyoung;Jang, Jungim;Park, Sihyeon;Jung, Hosung
    • Molecules and Cells
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    • v.45 no.11
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    • pp.846-854
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    • 2022
  • Neurons make long-distance connections via their axons, and the accuracy and stability of these connections are crucial for brain function. Research using various animal models showed that the molecular and cellular mechanisms underlying the assembly and maintenance of neuronal circuitry are highly conserved in vertebrates. Therefore, to gain a deeper understanding of brain development and maintenance, an efficient vertebrate model is required, where the axons of a defined neuronal cell type can be genetically manipulated and selectively visualized in vivo. Placental mammals pose an experimental challenge, as time-consuming breeding of genetically modified animals is required due to their in utero development. Xenopus laevis, the most commonly used amphibian model, offers comparative advantages, since their embryos ex utero during which embryological manipulations can be performed. However, the tetraploidy of the X. laevis genome makes them not ideal for genetic studies. Here, we use Xenopus tropicalis, a diploid amphibian species, to visualize axonal pathfinding and degeneration of a single central nervous system neuronal cell type, the retinal ganglion cell (RGC). First, we show that RGC axons follow the developmental trajectory previously described in X. laevis with a slightly different timeline. Second, we demonstrate that co-electroporation of DNA and/or oligonucleotides enables the visualization of gene function-altered RGC axons in an intact brain. Finally, using this method, we show that the axon-autonomous, Sarm1-dependent axon destruction program operates in X. tropicalis. Taken together, the present study demonstrates that the visual system of X. tropicalis is a highly efficient model to identify new molecular mechanisms underlying axon guidance and survival.

Picropodophyllotoxin Inhibits Cell Growth and Induces Apoptosis in Gefitinib-Resistant Non-Small Lung Cancer Cells by Dual-Targeting EGFR and MET

  • Jin-Young, Lee;Bok Yun, Kang;Sang-Jin, Jung;Ah-Won, Kwak;Seung-On, Lee;Jin Woo, Park;Sang Hoon, Joo;Goo, Yoon;Mee-Hyun, Lee;Jung-Hyun, Shim
    • Biomolecules & Therapeutics
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    • v.31 no.2
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    • pp.200-209
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    • 2023
  • Patients with non-small-cell lung cancer (NSCLC) with epidermal growth factor receptor (EGFR) amplification or sensitive mutations initially respond to the tyrosine kinase inhibitor gefitinib, however, the treatment becomes less effective over time by resistance mechanism including mesenchymal-epithelial transition (MET) overexpression. A therapeutic strategy targeting MET and EGFR may be a means to overcoming resistance to gefitinib. In the present study, we found that picropodophyllotoxin (PPT), derived from the roots of Podophyllum hexandrum, inhibited both EGFR and MET in NSCLC cells. The antitumor efficacy of PPT in gefitinib-resistant NSCLC cells (HCC827GR), was confirmed by suppression of cell proliferation and anchorage-independent colony growth. In the targeting of EGFR and MET, PPT bound with EGFR and MET, ex vivo, and blocked both kinases activity. The binding sites between PPT and EGFR or MET in the computational docking model were predicted at Gly772/Met769 and Arg1086/Tyr1230 of each ATP-binding pocket, respectively. PPT treatment of HCC827GR cells increased the number of annexin V-positive and subG1 cells. PPT also caused G2/M cell-cycle arrest together with related protein regulation. The inhibition of EGFR and MET by PPT treatment led to decreases in the phosphorylation of the downstream-proteins, AKT and ERK. In addition, PPT induced reactive oxygen species (ROS) production and GRP78, CHOP, DR5, and DR4 expression, mitochondrial dysfunction, and regulated involving signal-proteins. Taken together, PPT alleviated gefitinib-resistant NSCLC cell growth and induced apoptosis by reducing EGFR and MET activity. Therefore, our results suggest that PPT can be a promising therapeutic agent for gefitinib-resistant NSCLC.