• Archives of Plastic Surgery
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• v.40 no.5
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• pp.517-521
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• 2013

Background Diabetes is characterized by chronic hyperglycemia, which can increase reactive oxygen species (ROS) production by the mitochondrial electron transport chain. The formation of ROS induces oxidative stress and activates oxidative damage-inducing genes in cells. No research has been published on oxidative damage-related extracellular superoxide dismutase (EC-SOD) protein levels in human diabetic skin. We investigated the expression of EC-SOD in diabetic skin compared with normal skin tissue in vivo. Methods The expression of EC-SOD protein was evaluated by western blotting in 6 diabetic skin tissue samples and 6 normal skin samples. Immunohistochemical staining was also carried out to confirm the EC-SOD expression level in the 6 diabetic skin tissue samples. Results The western blotting showed significantly lower EC-SOD protein expression in the diabetic skin tissue than in the normal tissue. Immunohistochemical examination of EC-SOD protein expression supported the western blotting analysis. Conclusions Diabetic skin tissues express a relatively small amount of EC-SOD protein and may not be protected against oxidative stress. We believe that EC-SOD is related to the altered metabolic state in diabetic skin, which elevates ROS production.

• Journal of the Korean Society of Radiology
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• v.8 no.6
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• pp.325-332
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• 2014

Metallic nanoparticles have attractive properties in biomedical applications such as diagnostics and therapeutics. Cross linked dextran coated iron oxide nanoparticles (SPIONs) and silica coated gadolinium oxide nanoparticles (SPGONs) have been synthesized as a radiosensitizer in the proton beam cancer therapy. The dextran and silicaused for the protective moieties on the SPIONs and SPGONs respectively. Size distributions of synthesized nanoparticles were confirmed 3~5 nm for SPIONs and 30~100 nm for SPGONs by transmission electron microscope (TEM). Cell survival fraction measurement and Western blot assay were performed to evaluate the radiosensitization effects of synthesized radiosensitizer. The calculated radiosensitization of SPIONs and SPGONs at 90 % cell death from the measured cell survival curves were 1.23 and 1.03 respectively. Western blotting results also show the same consistent results that the amount of released cytochrome c from mitochondria was considerably increased for the cancer cells taken up SPIONs.

• Asian Pacific Journal of Cancer Prevention
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• v.13 no.12
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• pp.6463-6468
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• 2012

Purpose: To evaluated the effect of the gambogic acid (GA), one of the effective components of Garcinia, in combination with a new multi-targeted oral medication, sunitinib (SU) on renal cancer cell proliferation in vitro and on tumor growth in vivo. Methods: After treatment with GA or SU, either alone or in combination, MTT and FACS analysis were used to examine cell viability and cycle distribution of the renal carcinoma cell lines 786-0 and Caki-1. Western blotting was employed to examine the expression of proteins related to the cell cycle and vascular formation. Furthermore, a xenograft model was applied to study the antitumor efficacy of SU or GA alone or in combination, with immunohistochemistry to detect expression of proteins related to xenograft growth and angiogenesis. Western blotting was used to examine NF-${\kappa}B$ signaling pathway elements in xenografts. Results: Treatment of 786-0 and Caki-1 cells with GA or SU resulted in decreased tumor cell proliferation, especially with joint use. Cells accumulated more strongly in the sub-G1 phase after joint treatment with GA and SU than treatment of GA and SU alone. Western blotting arrays showed 1 protein significantly upregulated, 2 proteins downregulated, and 2 proteins unchanged. Moreover, combined use of GA and SU inhibited the growth and angiogenesis of xenografts generated from Caki-1 significantly. Immunohistochemistry arrays showed downregulation of the expression of proteins promoting xenograft growth and angiogenesis, and Western blotting showed inhibition of the NF-${\kappa}B$ signaling pathway after treatment by GA alone and in combination with SU in xenografts. Conclusions: Our results show that the joint use of GA and SU can provide greater antitumor efficacy compared to either drug alone and thus may offer a new treatment strategy for renal cell carcinoma.

• Food Science of Animal Resources
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• v.23 no.1
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• pp.75-79
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• 2003

This study was carried out to detect the pro-inflammatory cytokines(IL-1, IL-6, TNF-a, IL-18) and IL-1 receptor accessory in mongolia mare's milk by western blotting. IL-1 and TNF-a were detected in 4 samples of mare's milk Proteins of 6 kD and 7 kD were bound to specific antibody against hIL-18. But, IL-l and TNF-a were not detectable in Difco skim milk IL-6 like factor of 60 kD was detected in both Difco skim milk and mare's milk. Also, IL-1 receptor accessory of 55 kD was detected in the mongolia mare's milk.

• The Plant Pathology Journal
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• v.32 no.5
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• pp.452-459
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• 2016

The genomic region of Grapevine fanleaf virus (GFLV) encoding the movement protein (MP) was cloned into pET21a and transformed into Escherichia coli strain BL21 (DE3) to express the protein. Induction was made with a wide range of isopropyl-${\beta}$-D-thiogalactopyranoside (IPTG) concentrations (1, 1.5, and 2 mM) each for duration of 4, 6, or 16 h. However, the highest expression level was achieved with 1 mM IPTG for 4 h. Identity of the expressed protein was confirmed by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) followed by Western blotting. The expressed 41 kDa protein was purified under denaturing condition by affinity chromatography, reconfirmed by Western blotting and plate-trapped antigen enzyme-linked immunosorbent assay (PTA-ELISA) before being used as a recombinant antigen to raise polyclonal antibodies in rabbits. Purified anti-GFLV MP immunoglobulines (IgGs) and conjugated IgGs detected the expressed MP and GFLV virions in infected grapevines when used in PTA-ELISA, double antibody sandwich-ELISA, and Western blotting. This is the first report on the production of anti-GFLV MP polyclonal antibodies and application for the virus detection.

• Korean Journal of Microbiology
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• v.40 no.4
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• pp.348-354
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• 2004

The expression and antibacterial. activity of recombinant human lactoferrin (hLf) was studied from meth­ylotrophic yeast Pichia pastoris. The gene encoding hLf, isolated from human breast cDNA library, was subcloned into the expression vector, pPIC3.5K under the control of AOX1 promoter. The gene was integrated into the host chromosome and was identified by Southern blotting. The expression of the integrated gene was investigated by RT-PCR, Northern blotting, SDS-PAGE and Western blotting. Discrete band corresponding to hLf was detected from the SDS-PAGE, which was confirmed by Western blotting. The expression was also confirmed by RT-PCR and Northern blotting. The antibacterial activity of the recombinant hLf (rhLf) was investigated using Staphy­lococcus aureus ATCC 6538P and Micrococcus flavus ATCC 10240 as test organisms. The rhLf showed strong antibacterial activities against the bacteria. Furthermore, many Gram-negative animal pathogens such as E.coli ATCC8739, 25922, and Salmonella typhimurium 114 and 115, Pseudomonas fluorescens ID 963 I, P. aeruginosa KCCM 11802, and Gram-positive bacteria Bacillus mesentericus were also inhibited in their growth by the rhLf.

• Journal of Korean Orthopaedic Sports Medicine
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• v.4 no.1
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• pp.18-28
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• 2005

Purpose: The purpose of this study is to evaluate the clinical, radiological and histopathological results after microfracture surgery for degenerative arthritis of the knee. Materials and Methods: From Oct. 1997 to Dec. 1998, 48 knees in 46 patients were treated by microfracture technique. Their mean age at the time of operation was 56 years(range, 40-75 years) and mean period of follow-up study was one year(range, 7-20 months). For 24 knees in 22 patients, 'second-look' arthroscopies and biopsies were performed at 6 months following microfracture. At the last follow up clinical results were evaluated with Baumgaertner's scale. The specimens of 24 cases were stained with H-E, Safranin-O, and Masson's trichrome. Eighteen of 24 cases were stained immunohistochemically and the Western blotting test was performed on 12 cases for type II collagen. We analyzed the relationship of the Western blotting for type II collagen with clinical score, preoperative varus deformity, joint space widening in radiological result, extent of repaired articular cartilage in '2nd-look' arthroscopic findings, patient's age and weight. Results: Clinical results were excellent in 90% and good in 10%. Among the 24 knees, more than 80% of areas of chondral defect were covered with regenerated cartilage in 21 knees Histologically, the repaired tissue appears to be a hybrid of hyaline cartilage and fibrocartilage. Repaired cartilage contains variable amounts of type II collagen with immunohistochemical staining. The results of the Western blotting test were similar. The amounts of type II collagen formation had positive correlation with the extent of repaired cartilage and preoperative varus deformity. Conclusion: 'Second-look' showed that the chondral defect areas were covered with newly grown grayish white tissue. Articular cartilage repair was confirmed with histological and immunohisto-chemical study qualitatively, and the amount of type II collagen was calculated with the Western blotting test quantitatively. The exact nature and fate of repaired cartilagenous tissues need further long term follow-up study. The results of this study provide the rationale to select osteoarthritic patients indicated for microfracture surgery.

• Journal of Life Science
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• v.22 no.3
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• pp.318-323
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• 2012

This study involved observation of the inhibitory effect of 70% EtOH and water extracts from Doinseunggitang on melanin synthesis, tyrosinase activity, and western blotting using B16F10 melanoma cells. Doinseunggitang extracts inhibited melanin synthesis and tyrosinase activity in a dependent manner. As a result, it was found that Doinseunggitang 70% EtOH extracts inhibit melanin synthesis and tyrosinase activity, respectively, by 40% and 51%. In addition, western blotting analysis showed that 70% EtOH extracts inhibited tyrosinase, MITF, TRP-1, and TRP-2 expression. These results show that 70% ethanol extracts of Doinseunggitang could be developed as a skin whitening material in cosmetics.

• KSBB Journal
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• v.17 no.6
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• pp.582-585
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• 2002

In an effort to use plant biotechnology for the production of biopharmaceuticals, pollens collected from lily (Lilium longiflorum) were grown in vitro and transformed with a PCR-amplified 1.7 kb cDNA encoding human tissue plasminogen activator (tPA) using Agrobacterium via a vacuum infiltration process. Western blotting showed that transgenic lily pollen tubes selected on kanamycin for 16 hrs expressed a tPA protein with a size similar to the human standard, suggesting their possible use as a disposable host for rapid foreign protein production.

• Interdisciplinary Bio Central
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• v.4 no.2
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• pp.5.1-5.9
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• 2012

Small peptide tags such as c-myc, HA, or FLAG tag have facilitated efficient Western-blotting of proteins of interest especially when specific antibodies for the proteins are not available. However, the conventional Western-blotting requires the multi-steps process taking at least several hours up to two days. With examples of various applications, here we show a convenient and time-saving method for protein detection which employs a fluorescent chemical BDED and its binding peptide RC-tag. And we propose "Estern staining", as a standard term for protein detection method using fluorescent chemicals and their binding small peptide tags. Eastern staining may substitutes for the time-consuming "immuno-staining" in many versatile applications.