• Korean Journal of Veterinary Service
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• v.35 no.2
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• pp.153-157
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• 2012

A 7-year-old, castrated male dachshund dog with tumor of the spleen portion was referred to the Kangwon National University. The tumor removed surgically and tumor size was 6~7 cm. Histopathologically, this neoplasm was the presence of perivascular whorls of fusiform cells. The cells also arranged in interlacing bundles or storiform patterns. And this neoplasm consisted of spindle cells that often formed distinct whorls around a central capillary. Immunohistochemical analysis revealed multi-focally immunoreactivity for ${\alpha}$-smooth muscle actin, whereas not immunoreactive for desmin, S-100, von Willebrand factor. On the base of the histological and immunohistochemical results, this neoplasm was diagnosed as a canine hemangiopericytoma.

• Bulletin of the Korean Chemical Society
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• v.33 no.9
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• pp.2907-2909
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• 2012

A disintegrin and metalloprotease with thrombospondin domains (ADAMTS) are a member of peptidase and involved in processing of von Willebrand factor as well as cleavage of aggrecan, versican, brevican and neurocan. Among 19 subfamilies of human ADAMTS, ADAMTS-4 is a zinc-binding metalloprotease and is a famous therapeutic target for arthritis. It has been reported that a flavonoid luteolin shows inhibitory activity against ADMATS-4. In this study, we verified that luteolin is a potent inhibitor of ADAMTS-4 and probed the molecular basis of its action. On the basis of a docking study, we proposed a binding model between luteolin and ADAMTS-4 in which 3',4'-dihydroxyl groups in luteolin formed hydrogen bonding with ADMATS-4 and these interactions are important for its inhibitory activity against ADAMTS-4.

• The Korean Journal of Physiology and Pharmacology
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• v.19 no.1
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• pp.35-42
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• 2015

In cardiovascular disorders, understanding of endothelial cell (EC) function is essential to elucidate the disease mechanism. Although the mouse model has many advantages for in vivo and in vitro research, efficient procedures for the isolation and propagation of primary mouse EC have been problematic. We describe a high yield process for isolation and in vitro culture of primary EC from mouse arteries (aorta, braches of superior mesenteric artery, and cerebral arteries from the circle of Willis). Mouse arteries were carefully dissected without damage under a light microscope, and small pieces of the vessels were transferred on/in a Matrigel matrix enriched with endothelial growth supplement. Primary cells that proliferated in Matrigel were propagated in advanced DMEM with fetal calf serum or platelet-derived serum, EC growth supplement, and heparin. To improve the purity of the cell culture, we applied shearing stress and anti-fibroblast antibody. EC were characterized by a monolayer cobble stone appearance, positive staining with acetylated low density lipoprotein labeled with 1,1'-dioctadecyl-3,3,3',3'-tetramethyl-indocarbocyanine perchlorate, RT-PCR using primers for von-Willebrand factor, and determination of the protein level endothelial nitric oxide synthase. Our simple, efficient method would facilitate in vitro functional investigations of EC from mouse vessels.

• Journal of Veterinary Clinics
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• v.37 no.6
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• pp.336-338
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• 2020

A 12-year-old female spayed miniature schnauzer with one month history of halitosis had a cylindrical protruding mass about 2 × 2 × 1 cm in size arising from the right palatine tonsil. Histopathologically, tonsillar mass was covered with stratified squamous epithelium and many lymphoid follicles with germinal center were existed in the upper area of submucosa. Numerous variable sized, occasionally cystic dilated vascular channels (thin walled lymphatic channels) lined by flattened, discontinous endothelial cells were widely distributed throughout the core of fibrovascular stroma. According to immunohistochemistry (IHC), lining endothelial cells in vascular channels showed strong immunoreactivities for CD31 and von Willebrand factor. Based on the gross, histopathologic, and IHC findings, the oral mass of dog was diagnosed as tonsillar lymphangiomatous polyp, the second case in a veterinary field.

• Journal of Veterinary Clinics
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• v.40 no.5
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• pp.330-335
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• 2023

Platelet function evaluation by PFA-100 or -200 has been known to be objective and sensitive for assessing platelet function and dysfunction of Von Willebrand Factor in humans and dogs. However, using the C/EPI cartridge in dogs is controversial. This study aimed to establish a reference range for PFA closure time in healthy small breed dogs (body weight < 10 kg) and to evaluate the effectiveness of both C/ADP and C/EPI cartridges for these dogs. Citrated blood samples were collected from 50 clinically healthy small breed dogs that were admitted for presurgical procedures or health checkups, and closure times were measured using the PFA-200. Reference ranges were determined as 42-144 s (median 67 s, mean 71.2 s, SD ± 21.2 s, 95% RI 43-140 s) , for CT-C/ADP and 41-200 s (median 87, mean 91.2 s, SD ± 31.8 s, 95% RI 44-195 s) for CT-C/EPI. The present study demonstrated that the reference ranges for PFA closure times in small breed dogs are in line with existing reference ranges. The utilization of C/ADP cartridges is the preferred choice for evaluating platelet function in small breed dogs. However, due to variable responses of epinephrine to platelet aggregation in dogs, caution should be exercised when using C/EPI cartridges.

• Korean Journal of Food Science and Technology
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• v.29 no.4
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• pp.817-822
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• 1997

Inhibitory mechanism of the anticoagulant polysaccharide purified from the fruit body of Coriolus versicolor was investigated in this paper. The anticoagulant polysaccharide (CV-40-Va-1) was proposed to have functions of the inhibition of intrinsic pathway in the blood coagulation pathway together with the interuption of a human platelet aggregation induced by von Willebrand factor (vWF). CV-40-Va-I inhibited other factors of the coagulation cascade such as factor VIII, IX, and as well as thrombin. Especially, CV-40-Va-I inhibited the fibrin formation mediated by thromin, however the polysaccharide did not affect the fibrin formation directly but affected the anticoagulant activity through the activation of antithrombin III. The sulfation of the anticoagulant polysaccharide increased the anticoagulant activity, showing that the sulfate concentration of anticoagulant polysaccharide was important factor in the blood coagulation cascade. Low molecular weight subfraction (MW 1,000) obtained by partial hydrolysis of the CV-40-Va-1 generated potent antiplatelet activity, but showed decreased anticoagulant activity.

• Journal of Animal Science and Technology
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• v.50 no.6
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• pp.753-762
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• 2008

The primers for RT-PCR and RACE-PCR were designed by aligning the pig genomic sequence and the human complement factor B(CFB) coding sequence(CDS) from the GenBank. Each PCR product was amplified in pig cDNA and sequencing was carried out. The CDS length of pig CFB gene was determined to be 2298 bp. In addition, the pig CDS was more longer than human and mouse orthologs because of insertion and deletion. The identities of porcine nucleotide sequences with those of human and mice were 84% and 80%, and the identities of amino acids were 79% to 77%, respectively. Three complement control protein(CCP) domains, one Von Willebrand factor A(VWFA) domain and a serine protease domain, that are revealed typically in mammals, were found in the pig CFB gene. Based on the CDSs determined, the primers were designed in intron regions for amplification of entire length of exons. In amplification and direct sequencing with genomic DNAs of six pig breeds, three cSNPs(coding single nucleotide polymorphisms) were identified and verified as missense mutations. Using the Multiplex-ARMS method, we genotyped and verified the mutations identified from direct sequencing. To demonstrate recrudescence, we performed both direct sequencing and Multiplex-ARMS with two randomly selected DNA samples. The genotype of each sample exhibited the same results using both methods. Therefore, three cSNPs were identified from pig CFB gene and that can be used for haplotype analysis of the swine leukocyte antigen(SLA) class III region. Moreover, the results indicate that the Multiplex-ARMS method should be powerful for genotyping of genes in the SLA region.

• Biotechnology and Bioprocess Engineering:BBE
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• v.5 no.2
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• pp.130-135
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• 2000

The characteristics of two different modes of perfusion culture, intermittent and continuous bleedings, were investigated by culturing the hybridoma cells producing von Willebrand Factor (vWF) monoclonal antibody (McAb) in a 15 L bioreactor without clogging the filter. Both culture methods exhibited similar profiles of cell density and metabolite concentrations during the culture period at the cell concentration of around 1${\times}$107 cells/mL. When the perfusion rate was increased, the intermittrnt bleeding culture showed problems of ammonia accumulation and decrease of cell viability. The continuous bleeding culture in terms of nutrient consumption and metabolite production kinetics. But the analysis of specific oxygen consumption rate showed that the specific oxygen consumption rate of intermittent bleeding culture was similar to that of exponential growth phase. The continuous bleeding culture showed higher specific oxygen consumption rate of intermittent bleeding culture. finally we proved the possibility of long-term operation of continuous bleeding culture and produced approximately 40 g of vWF McAb in a 15L bioreactor after one-month operation.

• Korean Journal of Clinical Laboratory Science
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• v.41 no.1
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• pp.1-5
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• 2009

The Platelet Function Analyzer (PFA)$^{(R)}$-100 measures the ability of platelets activated in a high-shear environment to occlude an aperture in a membrane treated with collagen and epinephrine (CEPI) or collagen and ADP (CADP). The time taken for the flow across the membrane to stop (closure time, CT) is recorded. The aim of this study was to assess the potential of the PFA$^{(R)}$-100 as a primary clinical screening tool using the wide spectrum of clinical samples assessed for platelet function as well as to perform the optimal algorithm for the use of PFA$^{(R)}$-100. We established the reference interval in 460 hospital inpatients defined as having normal platelet function based on classical laboratory tests. The reference interval by using the range $5^{th}$ and $95^{th}$ percentile was 84~251 seconds for males CEPI-CT and 85~249 seconds for females CEPI-CT. A total of 1,200 inpatients were enrolled to identify impaired hemostasis before surgical interventions. The abnormal group showing prolonged CEPI-CT was 303 cases (18.9%). Only 3 cases had both abnormal CEPI-CT and CADP-CT. Several factors including sample errors, drugs, hematologic abnoralities were contributed to unexpected prolonged CEPI-CT for screening test. The von Willebrand factor (vWF:Ag) assay was performed only in one patient to verify the algorithm for the use of PFA$^{(R)}$-100. The PFA$^{(R)}$-100 was sensitive and rapid method for primary screening test of platelet dysfunction, so we can substitute it for the bleeding time in routine clinical practice.

• Proceedings of the Korean Society of Developmental Biology Conference
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• 2003.10a
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• pp.78-78
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• 2003

Adult stem cells can make identical copies of themselves for long periods of time. They also give rise to many differentiated mature cell types that have characteristic morphology and specialized function. Human adult stem cells are the attractive raw materials for the cell/tissue therapy, however, it is not easy to get from the adult tissues. In the present study, we tried to isolate a cell population derived from human umbilical cord vein which has been discarded after birth. The cells were isolated after treatment of the umbilical vein with collagenase or trypsin. After 3 days of culture, two kinds of cell populations were found consisting of adherent cells with endothelial cell-like and fibroblast-like morphology, respectively. When these cells were subcultured 12 times over a period of 3 months, almost cells appeared uniformly to exhibit fibroblastoid morphology which was different from that of mesenchymal stem cells obtained from human bone marrow The results of RT-PCR analyses showed distinct expression of BMP-4, oct-4, and SCF genes but not of GATA, PAX-6 and Brachyury genes. On immunohistochemical staining, the cells were negative for the von Willebrand factor(vWF), alpha-smooth muscle actin and placental alkaline phosphatase. From these observations, it is suggested that stem-like cells might be present in human umbilical cord vein.