• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2003년도 학술발표대회 발표논문초록집
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• pp.43-43
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• 2003

This study was conducted to produce transgenic pig harboring human tissue plasminogene activator (tPA) gene. Two different tPA genes containing bovine $\beta$-casein promoter and mouse uroplakin promoter were prepared for microinjection and confirmed the expression level of tPA protein from the CHO (Chinese hamster ovary) cell lines by gene transfection. Concentration of tPA expression from the six cell lines (all of CHO cells) were average 212.4 ng/ml. Reconstructed DNA to used the CHO cell were microinjected into the pronuclei of in vivo embryos The total of 2,307 zygotes were collected from 95 donors and 1,851 embryos were in 1-cell stage which were visualized the pronuclei for DNA microinjection. The concentration of linear DNA was 2.0 ng per microliter and injected into zygotes with two pronuclei on an inverted Nikon microscope equipped with narishige micromanipulator and modulation contrast optics. The 541 embryos injected with bovine $\beta$-casein promoter-tPA were transferred to 22 recipients. The 1,154 embryos injected with mouse uroplakin promoter-tPA were transferred to 51 recipients. Sixty nine offspring from 9 delivered sows were produced. We analysed the transgenes with PCR methods from 69 offsprings, but could not detect the PCR product from piglet tails DNA.

• Reproductive and Developmental Biology
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• 제34권3호
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• pp.185-191
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• 2010

Plasminogen activators (PAs) are serine protease that cleave plasminogen to form the active protease plasmin and may participate in mammalian fertilization. Although correlations have been reported between reactive oxygen species (ROS) and sperm function, the relationship between PA activity and ROS is unknown. We determined the effects of ROS on sperm function and PA activities in boar spermatozoa preincubated under the X-XO system. When spermatozoa were treated with the X+XO group, a significant increase (p<0.05) was observed in the percentage of acrosome reacted spermatozoa compared with that of the control group. However, when antioxidants were added to the medium with X+XO, the rate of acrosome reaction tended to decrease. Also, a significantly lower percentage of acrosome reacted spermatozoa was observed in the X+XO+catalase group at 6 hr of incubation compared with that of X+XO group. The density of malondialdehyde (MDA) was higher in the X+XO group than in different treatment groups. In another experiment, incubation of spermatozoa in medium with X+XO was associated with a significant (p<0.05) increase in activity of tPA-PAI and tPA compared with the control group. Antioxidants decreased the increased activity of tPA-PAI and tPA by preincubation in the X-XO system. Also, a significantly lower (p<0.05) activities of tPA-PAI and tPA were observed in the X+XO+catalase group compared with the X+XO group. No significant differences, however, were observed in the activity of uPA. These results suggest that the increase of acrosome reaction by the X-XO system resulted in increase of PAs activity in the sperm incubation medium.

• Biotechnology and Bioprocess Engineering:BBE
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• 제5권2호
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• pp.123-129
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• 2000

A high cell density culture system for the anchorage dependent CHO cells was developed based on the combination of in removal of ammonium ion and microcarrier culture system, and semi-fed-batch feeding of glucose and glutamine was employed to the developed culture system. The glass bead was selected as an optimum microcarrier in terms of cell growth. An ammonium ion selective zeolite, Phillipsite-Gismondine, was packed in a dialysis menium ion. The semi-fed-batch operation was employer to the novel culture system for the high density cell culture, and the results showed the cell growth was improved by 32% and tPA productivity by 250%.

• 한국독성학회:학술대회논문집
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• 한국독성학회 1997년도 국제 심포지움 및 춘계 학술대회
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• pp.57-60
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• 1997

Aloe의 추출 시료들 중 혈관생성을 촉진하는 물질이 G1M1D1M1 분획에 있음을 chorioallantoic-membrane(CAM) assay, in vitro angiogenesis assay, u-PA, PAI, u-PAR, 및 matrix metalloprotease(MMP)의 유전자 발현조사, gelatin zymogram assay, modified CAM assay를 하여 확인 하였다. 그런 다음 G1M1D1M1 분획을 다시 분리하여 단일 성분인 NY932와 3종의 분획들을(U>3,000, 3,000>U>1,000, U<1,000) 얻었다. 이들 각각에 대한 혈관 생성 촉진 작용을 CAM assay 방법으로 조사한 결과 순수단일 성분인NY932가 혈관 생성 촉진 활성을 가장 높게 나타났다. 즉 NY932의 농도를 100ng, 2, 10, 35, 60 $\mu$g으로 투여하였을 때, 각각 2/18($11\%$), 4/10($49\%$), 16/22($73\%$), 15/17($88\%$), 9/9($100\%$)의 혈관생성 촉진 효과를 보였다. 따라서 순수 단일 성분인 NY932의 혈관 생성 촉진 작용기전을 규명하기 위하여 혈관 생성에 관련된 효소들[matrix metalloprotease(MMP)s, urokinase-plasminogen activator(uPA) 등]과 그 저해제들(TIMPs, PAI)의 유전자 발현을 조사하였으며, wounding migration assay등을 수행하였다.

• Journal of Microbiology and Biotechnology
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• 제13권6호
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• pp.989-992
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• 2003

Recombinant human tissue plasminogen activator deletion mutein (Reteplase) is a clinically promising thrombolytic drug. Reteplase cDNA was subcloned into a bacteria expression system, and the resultant recombinant was biologically characterized. The Reteplase was expressed in Escherichia coli as an inclusion body, and the downstream processes of the Reteplase inclusion body included denaturation, renaturation, and purification. A protein disulfide isomerase (PDI) was used to assist the refolding of Reteplase, and it was found to increase the refolding rate from less than 2％ to more than 20％. The refolded Reteplase was purified through two chromatography steps, including lysine-coupled agarose affinity chromatography and then CM-sepharose cation-exchange chomatography. The purity of r-PA was analyzed by Western bolt analysis, and N-terminal amino acid and amino acid composition analyses confirmed the end-product. Reteplase showed higher thrombolytic potency in an animal thrombus model.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2887-2889
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• 2012

Malignant tumors have a capacity to degrade the extracellular matrix by controlled proteolysis. One system involved in these processes is the urokinase-type plasminogen activator (uPA) system. uPAR levels are elevated in tumors from several types of cancer. Our study was planned to investigate serum and urine levels of uPAR in breast cancer patients (n=180) and healthy controls (n=60) by ELISA. Serum (p<0.001) and urine (p<0.001) uPAR values in the patients were both significantly elevated. High serum and urine levels of uPAR can be used as diagnostic tools in lymph node positive patients.

• Asian Pacific Journal of Cancer Prevention
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• 제13권6호
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• pp.2437-2444
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• 2012

Tumor metastasis remains the principal cause of treatment failure and poor prognosis in patients with colorectal cancer. It is a multistage process which includes proteolysis, motility and migration of cells, proliferation in a new site, and neoangiogenesis. A crucial step in the process of intra- and extra-vasation is the activation of proteolytic enzymes capable of degrading the extracellular matrix (ECM). In this stage, urokinase plasminogen activator receptor (uPAR) and matrix metalloproteinases (MMPs) are necessary. Micrometastases need the presence of growth factor and vascular growth factor so that they can form macrometastasis. In addition, cell adhesion molecules (CAMs) and guanine nucleotide exchange factors (GEFs) play important roles in the progression of colorectal cancer and metastatic migration. Further elucidation of the mechanisms of how these molecules contribute will aid in the identification of diagnostic and prognostic markers as well as therapeutic targets for patients with colorectal metastasis.

• BMB Reports
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• 제47권4호
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• pp.227-232
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• 2014

Histone deacetylase-3 (HDAC3) is involved in cellular proliferation, apoptosis and transcriptional repression. However, the role of HDAC3 in angiogenesis remains unknown. HDAC3 negatively regulated the expression of angiogenic factors, such as VEGF and plasminogen activator inhibitor-1 (PAI-1). HDAC3 showed binding to promoter sequences of PAI-1. HDAC3 activity was necessary for the expression regulation of PAI-1 by HDAC3. VEGF decreased the expression of HDAC3, and the down-regulation of HDAC3 enhanced endothelial cell tube formation. HDAC3 negatively regulated tumor-induced angiogenic potential. We show the novel role of HDAC3 as a negative regulator of angiogenesis.

• 한국임상수의학회지
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• 제32권1호
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• pp.130-134
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• 2015

A 13-year-old, castrated male Yorkshire terrier was presented with acute blepharospasm 2 months after phacoemulsification with intraocular lens implantation in the left eye. Menace response was weakly positive in the left eye. Intraocular pressure (IOP) was 98 mmHg in the left eye. On ophthalmic examination using slit lamp biomicroscope, episcleral injection and corneal edema were observed in the left eye. Aqueous flare (1+) and iris bombe with peripheral anterior synechia were also detected in the left eye. In spite of medical treatments, IOP was remained more than 50 mmHg. Synechiolysis and radial iridectomy with irrigation/aspiration of anterior chamber were performed in the left eye. One week after the surgery, tissue plasminogen activator was injected to resolve the blood and fibrin clot in the left anterior chamber. Menace response was recovered completely 1 month after the surgery and IOP was maintained less than 20 mmHg more than 6 months.

• The Korean Journal of Physiology and Pharmacology
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• 제10권5호
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• pp.223-229
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• 2006

The emerging concept of the 'neurovascular unit' may enable a powerful paradigm shift for neuroscience. Instead of a pure focus on the 'neurobiology' of disease, an opportunity now exists to return to a more integrative approach. The neurovascular unit emphasizes that signaling between vascular and neuronal compartments comprise the basis for both function and dysfunction in brain. Hence, brain disorders are not just due to death of neurons, but instead manifested as cell signaling perturbations at the neurovascular interface. In this mini-review, we will examine 3 examples of this hypothesis: neurovascular mechanisms involved in the thrombolytic therapy of stroke, the crosstalk between neurogenesis and angiogenesis, and the link between vascular dysfunction and amyloid pathology in Alzheimer's disease. An understanding of cell-cell and cell-matrix signaling at the neurovascular interface may yield new approaches for targeting CNS disorders.