• Korean Circulation Journal
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• v.53 no.6
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• pp.351-366
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• 2023

Along with the development of immunosuppressive drugs, major advances on xenotransplantation were achieved by understanding the immunobiology of xenograft rejection. Most importantly, three predominant carbohydrate antigens on porcine endothelial cells were key elements provoking hyperacute rejection: α1,3-galactose, SDa blood group antigen, and N-glycolylneuraminic acid. Preformed antibodies binding to the porcine major xenoantigen causes complement activation and endothelial cell activation, leading to xenograft injury and intravascular thrombosis. Recent advances in genetic engineering enabled knock-outs of these major xenoantigens, thus producing xenografts with less hyperacute rejection rates. Another milestone in the history of xenotransplantation was the development of co-stimulation blockaded strategy. Unlike allotransplantation, xenotransplantation requires blockade of CD40-CD40L pathway to prevent T-cell dependent B-cell activation and antibody production. In 2010s, advanced genetic engineering of xenograft by inducing the expression of multiple human transgenes became available. So-called 'multi-gene' xenografts expressing human transgenes such as thrombomodulin and endothelial protein C receptor were introduced, which resulted in the reduction of thrombotic events and improvement of xenograft survival. Still, there are many limitations to clinical translation of cardiac xenotransplantation. Along with technical challenges, zoonotic infection and physiological discordances are major obstacles. Social barriers including healthcare costs also need to be addressed. Although there are several remaining obstacles to overcome, xenotransplantation would surely become the novel option for millions of patients with end-stage heart failure who have limited options to traditional therapeutics.

• Journal of Veterinary Clinics
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• v.14 no.1
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• pp.30-36
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• 1997

It is well estabilished that IL-4, IL-6, $IFN{\gamma}$ and immunoglobulin production are increased as a consequence of advancing age, and endothelial cells increase their expression of pro-inflammatory surface protein, such as ICAM-1, as a consequence of contact with inflammatory cytokines, including IL-1, $TNF{\alpha} or INF{\gamma}.$ This study was designed to define the relationship between age-associated changes of cytokines and immunoglobulin production and ICAM-1 expression in aged mice. Serum from aged mice had elevated IL-6 and immunoglobulin levels compared to mature adult controls and activation-induced production of $IFN{\gamma}$ of splenocytes from aged mice were increased above normal adult level. By immunoperoxidase staining, ICAM-1 in hearts of normal adult mice was expressed occasionally at low levels, but in aged mice, the number of positive blood vessels for ICAM-1 were increased and also the immunoreactivity were stronger. Therefore, these finding indicate that the altered expression of ICAM-1 eith normal aging may actually be correlative to these age-related changes in cytokiness and immunoglobulin production.

• Korean Journal of Agricultural Science
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• v.48 no.2
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• pp.353-365
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• 2021

Previously, we reported that tissue factor (Tf) was included in the list of differentially expressed genes as an upregulated gene in a rejected porcine heart after xenotransplantation into monkey. In this study, we analyzed that expression of Tf in aortic endothelial cells (pAEC) isolated from alpha 1,3-galactosyltransferase knockout pig in response to allogeneic porcine serum and xenogeneic human serum. The consequence was significant upregulation of Tf expression by responding to human serum compared with porcine serum. To analyze the function of Tf gene as a promoter, we constructed reporter vectors for expression of luciferase linked to 1,246 and 787 base pairs of porcine Tf (pTF1246 and pTF787), and 535 base pairs of human TF (hTF535) sequences including putative promoter regions and AP-1 biding site at the 5' end. The reporter vectors were transfected into pAEC including cytomegalovirus enhancer/chicken β-actin (CAG)-luciferase vector as a control. Luciferase assay showed that all of the promoters were insufficient to express luciferase compared with CAG promoter in basic culture conditions. Notably, pTF1246, pTF787, and hTF535 led to a significant increase of luciferase expression in response to human serum compared with porcine serum while no change of CAG. pTF1246 and pTF787 showed higher expression than hTF535. Taken together, our findings suggest that pTF1246 and pTF787 promoters could mediate target gene expression specifically at xenogeneic stress condition.

• Biomedical Science Letters
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• v.18 no.4
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• pp.420-427
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• 2012

Sprague-Dawley (SD) rats were orally administered with NG-nitro-L-arginine methyl ester (L-NAME), which inhibits or blocks the production of nitric oxide from L-arginine in vascular endothelial cells and vessel tissue. We examined the effects of nitric oxide on some physiological changes such as blood pressure and heart rate, and confirms the apoptosis induced by the suppressed nitric oxide activity in the kidney. This study was performed to investigate correlation between the activities of nitric oxide and apoptosis by immunohistochemical changes of apoptotic control proteins with regulated chronic inhibition of nitric oxide. In the kidney from L-NAME-treated group, immunohistochemical reaction to the antigens of apoptosis inhibiting proteins such as bcl-2 and bcl-xL, exhibited a time-dependent reduction. The expression of apoptosis-inhibiting proteins such as bax and p53 increased expression in proportion to the duration of treatment. The most sensitive apoptosis regulating proteins to L-NAME were p53 in stimulation and bcl-2 in inhibition, respectively.

• Korean Journal of Food Science and Technology
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• v.46 no.3
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• pp.375-383
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• 2014

This study aimed to investigate the effects of 50% ethanol extract of ripe black raspberry (Rubus occidentalis, RBR) on hypertension in human umbilical vein endothelial cells (HUVECs) and in spontaneously hypertensive rats (SHR). Angiotensin converting enzyme (ACE) inhibition and activation of nitric oxide production by endothelial nitric oxide synthase were significantly regulated by RBR in HUVEC cells. Moreover, the SHR showed significantly higher levels of blood pressure, ACE, renin, endothelin-1, and interleukin-6 than Wistar Kyoto rats (WKY). However, treatment with captopril and RBR decreased the levels of these hypertension-related events in the SHR. The renal arteriole showed greater media thickness/lumen diameter (%) in the SHR than in the WKY. However, media thickness/lumen diameter (%) was reduced in SHR by treatment with captopril and RBR. In addition, the number of eosinophilic cardiac muscle cells was decreased in the heart muscles after treatment with captopril and RBR. Therefore, this study suggests that 50% ethanol extract of RBR may be useful for the prevention and treatment of high blood pressure.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.164-166
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• 2003

Despite recent advances in the treatment of acute myocardial infarction, the ability to repair extensive myocardial damage is limited. Revascularization in ischemic myocardium is required to improve cardiac function and prevent further scar tissue formation. Bone marrow contains endothelial precursors that can be used to induce neovascularization in ischemic myocardium. To develop a new therapy for myocardial infarction, we investigated if implantation of bone marrow mononuclear cells (BM-MNCs) using biodegradable matrices could enhance neovascularization in ischemic myocardium. Eight weeks after implantation, the damaged myocardium implanted with BM-MNCs and fibrin gels exhibited significantly greater angiogenic responses than those implanted with either fibrin gels or BM-MNCs alone. Fibrin gels disappeared completely 8 weeks after implantation. Echocardiography revealed improved heart functions. These results suggest that implantation of BM-MNCs using fibrin gel matrix efficiently induces neovascularization and improved heart functions in ischemic myocardium.

• Journal of Chest Surgery
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• v.37 no.2
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• pp.131-138
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• 2004

High-flow gas insufflation to get a bloodless field during off-pump coronary artery bypass may have adverse effects on the coronary endothelium. This study was designed (1) to elucidate the effect of carbon dioxide gas insufflations on the coronary endothelium at different flow rates and (2) to assess the protective effect of humidifcation against the coronary endothelial damage. Material and Method: In nine pigs, the left anterior descending coronary artery (LAD) was exposed after a median sternotomy. The LAD was divided into 4 segments and a coronary arteriotomy was made in each LAD segment in the beating heart. The far distal arteriotomy was exposed to room air for 10 minutes and was harvested as a control. Non-humidified carbon dioxide gas at a continuous flow rate of 5 L/min (Group I), humidified carbon dioxide gas at a continuous flow rate of 5 L/min (Group II), and humidified carbon dioxide gas at a continuous flow rate of 10 L/min (Group III) were insufflated for 10 minutes on each coronary arteriotomy site, respectively. After harvesting the coronary segments, hematoxylin-eosin staining, elastic fiber staining, and immunostaining with a CD34 monoclonal antibody were performed to evaluate the depth of endothelial damage and to count the residual endothelial cells, Result: In all three groups (Group I, II, and III), internal elastic laminae were preserved, however, the endothelial layers were significantly damaged by carbon dioxide gas insufflation. The mean percentages of remaining endothelial cells were 20,9$\pm$16.7%, 39.3$\pm$19.6%, and 6.8$\pm$5.3%, in groups I, II, and III, respectively. The percentages of remaining cells were significantly higher in group II than in groups I and III (p=0.008). The percentages of remaining cells were significantly higher in group I than in group III (p=0.008). Conclusions: The harmful effect of carbon dioxide gas insufflation on the coronary endothelium was dependent on the flow rate. The addition of humidification did not protect the coronary endothelium from denudation injury caused by high flow carbon dioxide gas insufflations.

• Korean Journal of Microbiology
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• v.44 no.4
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• pp.282-288
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• 2008

Tissue ischemia resulting from the constriction or obstruction of blood vessels leads to an illness that may affect many organs including the heart, brain, and legs. In recent years, considerable progress has been made in the field of therapeutic angiogenesis and the new approaches are expected to cure those "no-option patients" who are unsuited to conventional therapies. Although single angiogenic growth factor may be successful in inducing angiogenesis, combination of multiple growth factors is increasingly sought these days to augment the therapeutic responses. This trend is proper in light of the fact that blood vessel formation is a complex and multi-step process that requires the actions of many different factors. To meet the growing need for functionally significant blood flow recovery in the ischemic tissues, a novel strategy that can provide concerted actions of multiple factors is required. One way to achieve such a goal is to use a transcription factor that can orchestrate the expression of multiple target genes in the ischemic region and thus induce significant level of angiogenesis. Here, a putative transcription factor, cardiac ankyrin repeat protein (CARP), was evaluated in adenoviral vector context for angiogenic activity in human umbilical vein endothelial cells. The results indicated significant increase in proliferation, capillary-like structure formation, and induction of vascular endothelial growth factor, a typical angiogenic gene. Taken together, these results suggest that CARP represents itself as a novel target for therapeutic angiogenesis and warrants further investigation.

• Journal of Chest Surgery
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• v.29 no.4
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• pp.373-380
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• 1996

Prevention of thromboembolism is the most important task in the development of bioconpatible small caliber artificial vascular graft. In normal vessels, vascular endothelial cells maintain homeosatsis by secreting numerous factors. The aim of this study is to develope a method which Improves biocompatibility of small caliver polyurethane graft using endothelial cell culture technique, and ev luate the efTectiveness of extracelluar matrix for endothelization which was produced by cultured fibroblast. Methods ; Multiporous polyurethane tube of 3 mm diameter, 0.3 mm thickness was manufactured for vascular graft. Three mongrel dogs were intubated and internal jugular veins removed. Extracelluar matrix produced by cultured flbrobast which was obtained from dog's internal jugular vein were coated to the polyurethane graft. Then, endothelial cells extracted from Jugular vein were cultured and fixed on the extracelluar matrix layer of vascular graft. Endothelial cell coated vascular grafts were implanted to the carotid arteries of experimental dogs as interposed autograft. Implanted grafts were removed after 3 and 6 weeks. As a control, PTFE graft was interposed on carotid artery. These experiments demonstrated that extracelluar matrix produced by fibroblast can afford a base for endothelial cell linings of polyurethane graft. Although thrombosis were developed on autografted en othelial cell coated graft, 33% opening was noticed, and showed less adhesion to adjacent tissue layer. These findings suggest that fiboblast produced extracelluar matrix which can be used for edothelial cell lining vascular graft, and by improving the cultured endothelial cell function, there will be a new modality for reducing thrombosis on small vascular graft.

• Journal of Korean Neurosurgical Society
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• v.63 no.2
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• pp.163-170
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• 2020

Objective : Milk fat globule-epidermal growth factor VIII (MFG-E8) may play a key role in inflammatory responses and has the potential to function as a neuroprotective agent for ameliorating brain injury in cerebral infarction. This study aimed to determine the role of MFG-E8 in brain injury in the subacute phase of cerebral ischemia in a rat model. Methods : Focal cerebral ischemia was induced in rats by occluding the middle cerebral artery with the modified intraluminal filament technique. Twenty-four hours after ischemia induction, rats were randomly assigned to two groups and treated with either recombinant human MFG-E8 or saline. Functional outcomes were assessed using the modified Neurological Severity Score (mNSS), and infarct volumes were evaluated using histology. Anti-inflammation, angiogenesis, and neurogenesis were assessed using immunohistochemistry with antibodies against ionized calcium-binding adapter molecule 1 (Iba-1), rat endothelial cell antigen-1 (RECA-1), and bromodeoxyuridine (BrdU)/doublecortin (DCX), respectively. Results : Our results showed that intravenous MFG-E8 treatment did not reduce the infarct volume; however, the mNSS test revealed that neurobehavioral deficits were significantly improved in the MFG-E8-treated group than in the vehicle group. Immunofluorescence staining revealed a significantly lower number of Iba-1-positive cells and higher number of RECA-1 in the periinfarcted brain region, and significantly higher numbers of BrdU- and DCX-positive cells in the subventricular zone in the MFG-E8-treated group than in the vehicle group. Conclusion : Our findings suggest that MFG-E8 improves neurological function by suppressing inflammation and enhancing angiogenesis and neuronal proliferation in the subacute phase of cerebral infarction.