• Journal of Embryo Transfer
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• v.30 no.3
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• pp.249-255
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• 2015

Diabetes mellitus, the most common metabolic disorder, is divided into two types: type 1 and type 2. The essential treatment of type 1 diabetes, caused by immune-mediated destruction of ${\beta}-cells$, is transplantation of the pancreas; however, this treatment is limited by issues such as the lack of donors for islet transplantation and immune rejection. As an alternative approach, stem cell therapy has been used as a new tool. The present study revealed that bone marrowderived mesenchymal stromal cells (BM-MSCs) could be transdifferentiated into pancreatic cells by the insertion of a key gene for embryonic development of the pancreas, the pancreatic and duodenal homeobox factor 1 (PDX1). To avoid immune rejection associated with xenotransplantation and to develop a new cell-based treatment, BM-MSCs from ${\alpha}$-1,3-galactosyltransferase knockout (GalT KO) pigs were used as the source of the cells. Transfection of the EGFP-hPDX1 gene into GalT KO pig-derived BM-MSCs was performed by electroporation. Cells were evaluated for hPDX1 expression by immunofluorescence and RT-PCR. Transdifferentiation into pancreatic cells was confirmed by morphological transformation, immunofluorescence, and endogenous pPDX1 gene expression. At 3~4 weeks after transduction, cell morphology changed from spindle-like shape to round shape, similar to that observed in cuboidal epithelium expressing EGFP. Results of RT-PCR confirmed the expression of both exogenous hPDX1 and endogenous pPDX1. Therefore, GalT KO pig-derived BM-MSCs transdifferentiated into pancreatic cells by transfection of hPDX1. The present results are indicative of the therapeutic potential of PDX1-expressing GalT KO pig-derived BM-MSCs in ${\beta}-cell$ replacement. This potential needs to be explored further by using in vivo studies to confirm these findings.

• Korean Journal of Veterinary Research
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• v.53 no.1
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• pp.37-42
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• 2013

Mesenchymal stem cells (MSCs) have ability to differentiate into multi-lineage cells, which confer a great promise for regenerative medicine to the cells. The aim of this study was to establish a method for isolation and characterization of adipose tissue-derived MSC (pAD-MSC) and bone marrow-derived MSC (pBM-MSC) in pigs. Isolated cells from all tissues were positive for CD29, CD44, CD90 and CD105, but negative for hematopoietic stem cell associated markers, CD45. In addition, the cells expressed the transcription factors, such as Oct4, Sox2, and Nanog by RT-PCR. pAD-MSC and pBM-MSC at early passage successfully differentiated into chondrocytes, osteocytes and adipocytes. Collectively, pig AD-MSC and BM-MSC with multipotency were optimized in our study.

• Proceedings of the Ginseng society Conference
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• 1978.09a
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• pp.67-74
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• 1978

The phenomenon of 'soil sickness' is one of the most important limiting factors for ginseng(Panax ginseng) production in Korea. The principal cause is known to be due to the root rots caused by Cylindrocarpon destructans and Fusarium solani. Attempts were made to control the root rots with non-polluting cultural methods or soil amendments. Among the nine soil amendments tested, crab shell, cow bone and pig feces were selected for further testing. Each of the three amendments increased the populations or various actinomycetes in the range of 10-25 times over that of non-amended soil, whereas the population of C. destructans was reduced to about $50-70\%$ as compared with the control. Five isolates of Streptomyces with clear zones on chitin-agar medium were selected and then tested for their antagonistic effects on C. destructans. When anyone of the five isolates of Streptomyces and C. destructans was grown together in a modified peptone broth, growth of the latter was highly inhibited. When three levels of crab shell, cow bone, or pig feces were used to amend potted soil infested with C. destruetans, the root rot ratings of ginseng seedlings were reduced to less than one half in all the treatments as compared to the control. In another similar experiment, crab shell and cow bone amendments resulted in almost complete control of the seedling root rots in soil infested with C. destructans or F. solani. In conclusion, biological control with soil amendments of ginseng root rots caused by C. destructans and F. solani was successful. Further basic studies should be pursued using soil amendments for better control. In addition, field experiments are needed to complement the soil amendment control measures in an integrated pest control program.

• Korean Journal of Soil Science and Fertilizer
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• v.44 no.5
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• pp.788-793
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• 2011

To evaluate fertilizer value of animal cadavers for agricultural recycling, fertilizer components of animal cadavers by pig and poultry were investigated using rendering and alkali (KOH) treatment methods. Total nitrogen concentrations in meat waste by pig and poultry using rendering treatment method were 7.80% and 9.30%, respectively. Total nitrogen concentration in meat waste of pig by KOH treatment method was lower than that by rendering treatment method. Organic matter concentrations in meat waste of pig and poultry ranged 87.8~97.4%. Total phosphorus concentrations in bone waste of pig using rendering and KOH treatment methods ranged 5.59~11.18%. Animal cadavers contains nitrogen, phosphorus, potassium and other nutrients essential to plant growth. The results of this study suggest that animal cadavers can supply some of the nutrient requirements of crops and is a valuable fertilizer as well.

• Biomolecules & Therapeutics
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• v.7 no.1
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• pp.97-104
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• 1999

A new antibiotic bead, CJ-40003 is a combination of three antibiotics, tobramycin, vancomycin and cefazolin embedded in bone cement, for the treatment of osteomyelitis. To evaluate the general pharmacological properties of CJ-40003, the effects of its active ingredients were investigated in mice, rats, dogs and isolated guinea pig ileum. The combination of three antibiotics (CA) did not affect general behavior, central nervous system, smooth muscles, gastrointestinal system, cardiovascular and respiratory system and water and electrolytes excretion when administered intravenously at the doses of 0.3, 1 and 3 mg/kg, respectively, into experimental animals. The CA had no effect on the contractile response of the isolated guinea pig ileum to various spasmogen at concentrations of 1, 3 and 10 $\mu\textrm{g}$/ml, respectively. In conclusion, the active ingredients of CJ-40003 showed no pharmacological effect in these studies.

• Maxillofacial Plastic and Reconstructive Surgery
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• v.30 no.5
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• pp.417-427
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• 2008

The aim of this study is to investigate the effect of anodizing surface to osseointegration of implant by using of resonance frequency analysis (RFA), quantitative and qualitative assessment of an anodically modified implant type with regard to osseous healing qualities. A total of 96 screw-shaped implants were prepared for this study. 72 implants were prepared by electrochemical oxidation with different ways. 24 (group 1 SP) were prepared at galvanostatic mode in 0.25M sulfuric acid and phosphoric acid. 24 (group 2GC) were prepared at galvanostatic mode in calcium glycerophosphate and calcium acetate and 24 (group 3 CMP (Calcium Metaphosphate) Coating were prepared at galvanostatic mode in 0.25M sulfuric acid and phosphoric acid followed by CMP coating. Rest of 24 (control group were as a control group of RBM surface. Bone tissue responses were evaluated by resonance frequency analysis (RFA) that were undertaken at 2, 4 and 6 weeks after implant placement in the mandible of mini-pig. Group 1 SP (anodized with sulfuric acid and phosphoric acid implants) demonstrated slightly stronger bone responses than control Group RBM. Group 2 GC (anodized surface with calcium glycerophosphate and calcium acetate implants) demonstrated no difference which were compared with control group. Group 3 GMP (anodized and CMP coated implants) demonstrated slightly stronger and faster bone responses than any other implants. But, all observation result of RF A showed no significant differences between experimental groups with various surface type. Histomorphometric evaluation demonstrated significantly higher bone-to-implant contact for group 2 GC. Significantly more bone formation was found inside threaded area for group 2 GC. It was concluded that group 2 GC (anodized surface with calcium glycerophosphate and calcium acetate implants) showed more effects on the bone tissue responses than RBM surface in initial period of implantation. In addition, CMP showed a tendency to promote bone tissue responses.

• The Journal of Korean Academy of Prosthodontics
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• v.46 no.6
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• pp.628-633
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• 2008

STATEMENT OF PROBLEM: The application of a simple, clinically applicable noninvasive test to assess implant stability are considered highly desirable. So far there is still a controversy about correlation of various tests and implant stability. PURPOSE: In order to assess implant stability, the development of a new method is critical. It's possible to assess implant stability by calculating energy and angular momentum during implant installation. The purpose of this study is to evaluate the correlation of energy and implant stability. MATERIAL AND METHODS: Twenty three implants were installed in two different types of pig bone. Type I bone was retrieved from the distal aspect of the rib, with more cortical bone. Type II bone came from a more proximal region with less cortical components and a higher content of bone marrow and spongeous trabeculae. Insertion torque, removal torque, ISQ values and angular momentum and energy were measured. Pearson Correlation test was done to analyze the relation between RFA, maximum insertion torque, mean insertion torque, bone type, energy and removal torque. RESULTS: Type I bone showed higher removal torque than type II bone. Energy value was significantly correlated with maximum insertion torque and mean insertion torque. RFA values were related with insertion torques but the significance was lower than Energy value. CONCLUSION: Within the limitation of this study energy values were considered clinically predictable method to measure the implant stability.

• Food Science of Animal Resources
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• v.32 no.2
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• pp.234-240
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• 2012

This study was conducted to examine the effects of low molecular weight gelatin hydrolysates (GH, less than 3kDa), extracted from pig skin collagen on the bone metabolism of ovariectomized (OVX) rats. The rats in the experimental groups were randomly segregated into six different treatment groups such as 1) NC, the normal rat fed AIN 93 diet (basal diet) only; 2) OC, the OVX rat fed the basal diet only; 3) GH 0.1, the OVX rat fed the basal diet with 0.1% GH; 4) GH 0.8, the OVX rat fed the basal diet with 0.8% GH; 5) G 0.1, the OVX rat fed the basal diet with 0.1% gelatin; 6) G 0.8, the OVX rat fed the basal diet with 0.8% gelatin. Body weight gain in the GH 0.1, GH 0.8, and G 0.8 was significantly higher than those in the NC and OC. Feed intake of the GH 0.1 and GH 0.8 was higher than that of the NC and OC, while no significant difference was found in feed efficiency ratio (FER). BMD of the GH 0.8 was higher than that of the OC. However, gelatin hydrolysates and gelatin resulted in higher BMC level compare to the OC. Serum HDL-cholesterol of rat fed GH and gelatin was higher than that of OC (p<0.05). LDL-C of the GH 0.1 and the GH 0.8 tended to be less than that of OC. Serum alkaline phosphatase (ALP) of the GH 0.1 was lower than that of the OC. The serum of GH 0.8 showed lower osteocalcin value than the OC (p<0.05). In addition, GOT and GPT levels significantly decreased in all treatment groups. These results indicated that gelatin hydrolysates from pig skin gelatin hydrolysates enhanced BMD and serum biochemical parameters related to bone metabolism. Therefore, the gelatin hydrolysates could be used as a beneficial material to improve bone health.

• Journal of Embryo Transfer
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• v.28 no.3
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• pp.281-287
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• 2013

A major barrier to progress in pig to primate organ transplantation or cell therapy is the presence of terminal ${\alpha}$-1,3-galactosyl epitopes on the surface of pig cells. Therefore, the purpose of this experiment was to establish and cha- racterize mesenchymal stromal/stem cells (MSCs) derived from ${\alpha}$-1,3-galactosyltransferase (GalT) knock out (GalT KO) pig to confirm their potential for cell therapy. Bone marrow (BM)-MSCs from GalT KO pig of 1 month old were isolated by Ficoll-Paque PLUS gradient and cultured with A-DMEM + 10% FBS on plastic dishes in 5% $CO_2$ incubator at 38.5. GalT KO BM-MSCs were analyzed for the expression of CD markers ($CD45^-$, $29^+$, $90^+$ and $105^+$) and in vitro differentiation ability (adiopogenesis and osteogenesis). Further, cell proliferation capacity and cell aging of GalT KO BM-MSCs were compared to Wild BM-MSCs by BrdU incorporation assay (Roche, Germany) using ELISA at intervals of two days for 7 days. Finally, the cell size was also evaluated in GalT KO and Wild BM-MSCs. Statistical analysis was performed by T-test (P<0.05). GalT KO BM-MSCs showed fibroblast-like cell morphology on plastic culture dish at passage 1 and exhibited $CD45^-$, $29^+$, $90^+$ and $105^+$ expression profile. Follow in ginduction in StemPro adipogenesis and osteogenesis media for 3 weeks, GalT KO BM-MSCs were differentiated into adipocytes, as demonstrated by Oilred Ostaining of lipid vacuoles and osteocytes, as confirmed by Alizarinred Sstaining of mineral dispositions, respectively. BrdU incorporation assay showed a significant decrease in cell proliferation capacity of GalT KO BM-MSCs compared to Wild BM-MSCs from 3 day, when they were seeded at $1{\times}10^3$ cells/well in 96-well plate. Passage 3 GalT KO and Wild BM-MSCs at 80% confluence in culture dish were allowed to form single cells to calculate cell size. The results showed that GalT KO BM-MSCs($15.0{\pm}0.4{\mu}m$) had a little larger cell size than Wild BM-MSCs ($13.5{\pm}0.3{\mu}m$). From the above findings, it is summarized that GalT KO BM-MSCs possessed similar biological properties with Wild BM-MSCs, but exhibited a weak cell proliferation ability and resistance to cell aging. Therefore, GalT KO BM-MSCs might form a good source for cell therapy after due consideration to low proliferation potency in vitro.

• Asian-Australasian Journal of Animal Sciences
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• v.33 no.11
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• pp.1837-1847
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• 2020

Objective: To evaluate the pancreatic differentiation potential of α-1,3-galactosyltransferase knockout (GalTKO) pig-derived bone marrow-derived mesenchymal stem cells (BM-MSCs) using epigenetic modifiers with different pancreatic induction media. Methods: The BM-MSCs have been differentiated into pancreatic β-like cells by inducing the overexpression of key transcription regulatory factors or by exposure to specific soluble inducers/small molecules. In this study, we evaluated the pancreatic differentiation of GalTKO pig-derived BM-MSCs using epigenetic modifiers, 5-azacytidine (5-Aza) and valproic acid (VPA), and two types of pancreatic induction media - advanced Dulbecco's modified Eagle's medium (ADMEM)-based and N2B27-based media. GalTKO BM-MSCs were treated with pancreatic induction media and the expression of pancreas-islets-specific markers was evaluated by real-time quantitative polymerase chain reaction, Western blotting, and immunofluorescence. Morphological changes and changes in the 5'-C-phosphate-G-3' (CpG) island methylation patterns were also evaluated. Results: The expression of the pluripotent marker (POU class 5 homeobox 1 [OCT4]) was upregulated upon exposure to 5-Aza and/or VPA. GalTKO BM-MSCs showed increased expression of neurogenic differentiation 1 in the ADMEM-based (5-Aza) media, while the expression of NK6 homeobox 1 was elevated in cells induced with the N2B27-based (5-Aza) media. Moreover, the morphological transition and formation of islets-like cellular clusters were also prominent in the cells induced with the N2B27-based media with 5-Aza. The higher insulin expression revealed the augmented trans-differentiation ability of GalTKO BM-MSCs into pancreatic β-like cells in the N2B27-based media than in the ADMEM-based media. Conclusion: 5-Aza treated GalTKO BM-MSCs showed an enhanced demethylation pattern in the second CpG island of the OCT4 promoter region compared to that in the GalTKO BM-MSCs. The exposure of GalTKO pig-derived BM-MSCs to the N2B27-based microenvironment can significantly enhance their trans-differentiation ability into pancreatic β-like cells.